EA044500B1 - NAPHTHYRIDINONE COMPOUNDS FOR USE AS T-CELL ACTIVATORS - Google Patents

Description

Related applications

This application claims priority under U.S. Provisional Patent Application No.

62/690444, filed June 27, 2018, the entire contents of which are incorporated into this application.

Technical field of the invention

The present invention generally relates to naphthyridinone compounds that activate T cells, stimulate T cell proliferation and/or have anticancer activity. This invention provides naphthyridinone compounds, compositions containing such compounds, and methods of using them. Additionally, the invention relates to pharmaceutical compositions containing at least one compound according to the invention, which can be used in the treatment of proliferative disorders such as cancer and viral infections.

BACKGROUND OF THE INVENTION

Human malignancies carry with them numerous genetic and epigenetic changes leading to the production of tumor antigens that are potentially recognized by the immune system (Sjoblom et al. (2006) Science 314:268-74). The adaptive immune system, consisting of T and B lymphocytes, has a powerful anticancer potential, characterized by broad capabilities and amazing specificity of response to a variety of tumor antigens. In addition, the immune system exhibits significant plasticity and immune memory. Successfully combining all these attributes of the adaptive immune system would make immunotherapy a unique approach among all cancer treatments. However, although an endogenous immune response to malignancy is observed in preclinical models and in patients, the response is ineffective and confirmed malignancies are treated as self and ignored by the immune system. Tumors can use several different mechanisms to actively undermine anticancer immunity, contributing to the development of a state of immune tolerance. These mechanisms include disruption of T cell signal transduction (Mizoguchi et al., (1992) Science 258:1795-98), suppression of regulatory cells (Facciabene et al., (2012) Cancer Res. 72:2162-71), and use of endogenous control cells. immune response points that down-modulate the intensity of adaptive immune responses and protect normal tissues from collateral damage by tumors to avoid immune damage (Topalian et al., (2012) Curr. Opin. Immunol. 24:1-6; Mellman et al. (2011) Nature 480:480-489).

Diacylglycerol kinases (DGKs) are lipid kinases that catalyze the conversion of diacylglycerol to phosphatidic acid and thereby terminate T cell functions via the TCR signaling pathway. Accordingly, DGKs serve as intracellular checkpoints, and inhibition of DGKs can be expected to stimulate T cell signaling pathways and T cell activation. Supporting data include studies using DGKa- or DGKZ-knockout mouse models that show a hyperreactive T-cell phenotype and improved anticancer immune activity (Riese M.J. et al., Journal of Biological Chemistry, (2011) 7: 5254-5265; Zha Y et al., Nature Immunology, (2006) 12:1343; Olenchock B.A. et al., (2006) 11:117481). In addition, tumor-infiltrating lymphocytes isolated from patients with renal cell carcinoma have been shown to overexpress DGKa, resulting in inhibition of T cell function (Prinz, P.U. et al., J. Immunology (2012) 12:5990-6000). Accordingly, DGKa and DGKZ are considered targets for cancer immunotherapy (Riese M.J. et al., Front Cell Dev Biol. (2016) 4:108; Chen, S.S. et al., Front Cell Dev Biol. (2016) 4:130; Avila- Flores, A. et al., Immunology and Cell Biology (2017) 95: 549-563; Noessner, E., Front Cell Dev Biol. (2017) 5: 16; Krishna, S., et al., Front Immunology ( 2013) 4:178; Jing, W. et al., Cancer Research (2017) 77:5676-5686.

There remains a need for compounds that can be used as inhibitors of one or both of DGKa and DGKZ. In addition, there remains a need for compounds that can be used as inhibitors of one or both of DGKa and DGKZ and have selectivity for DGKa and DGKZ over other diacylglycerol kinases, protein kinases and/or other lipid kinases.

Accordingly, an agent that is safe and effective in restoring T cell activation, lowering antigen threshold levels, enhancing anticancer functionality, and/or overcoming the suppressive effects of one or more endogenous immune checkpoints such as PD-1, LAG-3, and TGFe, would be a significant addition for the treatment of patients with proliferative disorders such as cancer as well as viral infections.

Applicants have found compounds that have inhibitory activity on one or both of DGKa and DGKZ. In addition, Applicants have found compounds that have inhibitory activity on one or both of DGKa and DGKZ and are selective for DGKa and DGKZ over other diacylglycerol kinases, protein kinases and/or other lipid kinases. It is proposed that these compounds can be used as pharmaceuticals having desirable values of stability, bioavailability, therapeutic index and toxicity, which are important for their commercial use as drugs.

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The essence of the invention

The present invention provides naphthyridinone compounds of formula (I), which can be used as inhibitors of DGKa, DGKZ or both DGKa and DGKZ, including salts and prodrugs of these compounds.

The present invention also provides pharmaceutical compositions containing a compound of formula (I), and/or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

The present invention also provides the use of a compound of formula (I), and/or a pharmaceutically acceptable salt thereof, for the treatment of a disease or disorder associated with the activity of DGKa, DGKZ, or both DGKa and DGKZ.

The compounds of formula (I) and compositions containing compounds of formula (I) can be used to treat, prevent or cure viral infections and various proliferative disorders such as cancer. Pharmaceutical compositions containing these compounds can be used to treat, prevent, or slow the progression of diseases or disorders related to various therapeutic areas, such as viral infections and cancer.

An expanded form of the above and other features of the present invention is given below in the description of the invention.

Detailed Description of the Invention

According to a first aspect of the present invention there is provided a compound of formula (I)

or a pharmaceutically acceptable salt thereof, wherein

R1 represents H, Cl, Br, -CN, C1-2 alkyl, -CH=CH2, -OCH3, -C(O)OH, -C(O)OCH3, -C(O)N(CH ₃ ) ₂ , -NH2 or -NHC(O)OC(CH ₃ ) ₃ ; R2 represents -CH ₃ , -CH2CH3 , -CH ₂ CH ₂ CH ₃ , -CH ₂ CH ₂ CH ₂ CH ₃ , -CH2CN, -CH2CH2CH2CN, -CH2CH2CF3, -CH ₂ CH=CH ₂ , -CH ₂ CH ₂ CH=CF ₂ , -CH ₂ C=CH, -CH2CH2OCH3, -CH2CH2CH2OCH3, -CH2CH2CH2OCH2CH2OCH3, -CH2CH2CH2C(O)CH3,

-CH2C(O)OCH2CH3, -CH2(cyclopropyl) or -CH2CH2(dioxanil);

R3 is H, F, Cl, Br, -CN, -CH3, -CF3, -NO2, -C(O)OCH2CH3 or -C(O)CF3;

R4 represents:

(a) 2,3-dihydro-1H-indenyl having from 1 to 2 substituents independently selected from F, -OH, -OCH3 and -OCH2CH=CH2; or (b) -CH2Ry, -C(CH3)2Ry, -CHRxRy, -CH2CH(OH)Rx, -CH(CH3)(CH2CH2OCH3) or cyclopropyl having a fluorophenyl substituent;

R _x represents C _1-2 alkyl, -CH(CH ₃ ) ₂ , -C(CH ₃ ) ₃ , -CH ₂ C(CH ₃ ) ₃ , -CH2OH, -CH2NH2, cyclopropyl, cyclobutyl, cyclohexyl or phenyl, having from 0 to 2 substituents independently selected from F, Cl, -OH and -OCH3; And

R _y is 1,3-benzodiazolyl, indazolyl, indolyl, ethyl indolyl, indolinyl, naphthalenyl, hydroxynaphthalenyl, oxoindolinyl, pyridinyl, methoxypyridinyl, pyrimidinyl or phenyl, having from 0 to 3 substituents independently selected from F, Cl, Br, - OH, -CN, -CH ₃ , -C(CH ₃ ) ₃ , -CHF ₂ , -CF3, -OCH ₃ , -OCF3, -OCH ₂ CH=CH ₂ , -OCH-C CH, -OCH ₂ (cyanopyridinyl ), -NH ₂ , -NHS(O) ₂ CH ₃ , -N(CH ₃ )(CH ₂ CH ₃ ), -N HC(O)CH ₃ , -NHC(O)O(C(CH ₃ ) ₃ ), -NHC(O)(phenyl), -NHC(O)NH(phenyl) and phenyl.

According to one embodiment, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt thereof, wherein R ₄ is -CH ₂ R _y , -CHRxRy or -CH2CH(OH)Rx.

According to one embodiment, the present invention provides a compound of formula (1), or a pharmaceutically acceptable salt thereof, wherein

R _x represents phenyl having from 0 to 2 substituents independently selected from F, Cl, -OH, C _1-2 alkyl, -CHF2, -OCH ₃ , -OCH ₂ CH=CH ₂ and -OCH-C CH; And

R _y represents phenyl having from 0 to 3 substituents independently selected from F, Cl, Br, -OH, -CN, C _1-4 alkyl, C _1-2 fluoroalkyl, C _1-2 alkoxy, C _1-2 fluoroalkoxy, -OSH ₂ CH=CH ₂ , -OSH ₂ C=CH, -OSH ₂ (cyanopyridinyl), -NRcRc, -NHS(O) ₂ CH ₃ , -N HC(O)(C _1-2 alkyl), -NHC(O)O(C _1-4 alkyl), -NHC(O)(phenyl), -NHC(O)NH(phenyl) and phenyl.

According to one embodiment, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt thereof, wherein

R4 is -CHRxRy;

R _x represents phenyl having from 0 to 2 substituents independently selected from F, Cl,

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-OH and -OSH ₃ ; And

R _y represents phenyl having from 0 to 3 substituents independently selected from F, Cl,

Br, -OH, -CN, -CH3, -C(CH3)3, -CHF ₂ , -CF3, -OCH3, -OCF3, -OCH ₂ CH=CH ₂ , -OCH2C CH, -OCH ₂ (cyanopyridinyl), -NH ₂ , -NHS(O)2CH3, -N(CH ₃ )(CH2CH ₃ ), -NHC(O)CH3, -NHC(O)O(C(CH3)3), -NHc(o)(phenyl ), -NHC(O)NH(phenyl) and phenyl.

According to one embodiment, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt thereof, wherein R4 is 2,3-dihydro1H-indenyl having from 0 to 2 substituents independently selected from F, Cl, - OH, C _1-2 alkyl, C _1-2 fluoroalkyl, C _1-2 alkoxy and -OCH ₂ CH=CH ₂ .

According to one embodiment, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt thereof, which is: ethyl 4-(4-(bis(4fluorophenyl)methyl)piperazin-1-yl)-6-bromo- 1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carboxylate (1); 6-bromo-4-(4-(2-hydroxybenzyl)piperazin-1 -yl)-1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3carbonitrile (2); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1 -yl)-1 -methyl-3-nitro-1,5-naphthyridin-2(1 H)one (3); 6-bromo-4-{4-[(4-fluorophenyl)[2-(prop-2-yn-1-yloxy)phenyl]methyl]piperazin-1-yl}-1-methyl-2oxo-1,2- dihydro-1,5-naphthyridine-3-carbonitrile (5); 6-bromo-4-{4-[(4-fluorophenyl)[2-(prop-2-yn-1-yloxy)phenyl]methyl]piperazin-1-yl}-1-methyl-2-oxo-1, 2-dihydro-1,5-naphthyridine-3-carbonitrile (5-7); 6bromo-4-{4-[(4-fluorophenyl)(2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile (8 -10); 8-{4-[(4-fluorophenyl)(2-hydroxyphenyl)methyl]piperazin-1-yl}-5methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2,7-dicarbonitrile (11 ); 6-bromo-4-{4-[(4-fluorophenyl)(2methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (1214); 6-bromo-4-{4-[(4-fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridine -3-carbonitrile (15-17); 8-{4-[(4-fluorophenyl)(2-methoxyphenyl)methyl]piperazin1-yl}-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (18-20); 6-bromo-4-[4-(6methoxy-2,3-dihydro-1 H-inden-1 -yl)piperazin-1 -yl]-1 -methyl-2-oxo-1,2-dihydro-1, 5-naphthyridine-3 carbonitrile (21); 6-bromo-4-{4-[1 -(4-fluorophenyl)ethyl]piperazin-1 -yl}-1 -methyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile (22- 24); 6-bromo-4-{4-[1-(4-fluorophenyl)propyl]piperazin-1-yl}-1-methyl-2oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (25- 27); 6-bromo-4-{4-[2-(4-fluorophenyl)-2-hydroxyethyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3 -carbonitrile (28-30); 6-bromo-4-{4[ 1 -(4-fluorophenyl)-2-hydroxyethyl]piperazin-1-yl} -1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3 carbonitrile (31); 8-{4-[1-(4-fluorophenyl)propyl]piperazin-1-yl}-5-methyl-7-nitro-6-oxo-5,6-dihydro1,5-naphthyridine-2-carbonitrile (32- 34); 6-bromo-4-{4-[cyclopropyl(4-fluorophenyl)methyl]piperazin-1-yl}1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (35) ; 8-{4-[2-(4-fluorophenyl)-2-hydroxyethyl]piperazin-1-yl}-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridin-2 -carbonitrile (36); 8-{4-[1(4-fluorophenyl)-2-hydroxyethyl]piperazin-1-yl}-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2carbonitrile ( 37); 1-methyl-4-{4-[(naphthalene-1-yl)methyl]piperazin-1-yl}-3-nitro-1,2-dihydro-1,5naphthyridin-2-one (38); 6-chloro-4-{4-[(4-fluorophenyl)[2-(prop-2-yn-1-yloxy)phenyl]methyl]piperazin-1yl}-1-methyl-2-oxo-1,2- dihydro-1,5-naphthyridine-3-carbonitrile (39); 8-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridin-2-carbonitrile (40 ); 8-(4benzhydrylpiperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridin-2-carbonitrile (41); 8(4-((2-hydroxyphenyl)(phenyl)methyl)piperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridin-2-carbonitrile ( 42); 8-(4-(1-(4-fluorophenyl)ethyl)piperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6dihydro-1,5-naphthyridine-2-carbonitrile (43- 45); 8-(4-(6-methoxy-2,3-dihydro-1H-inden-1-yl)piperazin1-yl)-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5 -naphthyridine-2-carbonitrile (46); 8-(4-(2-hydroxy-1phenylethyl)piperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (47); 8(4-(2-hydroxy-2-phenylethyl)piperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2carbonitrile (48); 8-(4-(cyclopropyl(4-fluorophenyl)methyl)piperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6dihydro-1,5-naphthyridine-2-carbonitrile (49-51 ); 4-(4-benzhydrylpiperazin-1-yl)-6-bromo-1-methyl-3nitro-1,5-naphthyridin-2(1H)-one (52); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1 -yl)-6-bromo-1 -methyl-3nitro-1,5-naphthyridin-2(1H)-one (53); 6-bromo-4-(4-(( 1 -ethyl-1H-indol-4-yl)methyl)piperazin-1 -yl)-1 methyl-3-nitro-1,5-naphthyridin-2(1H) -on (54); 6-bromo-1-methyl-4-(4-(naphthalene-1 -ylmethyl)piperazin-1 yl)-3-nitro-1,5-naphthyridin-2(1H)-one (55); 6-bromo-4-(4-((4-fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl)piperazin-1-yl)-1-methyl-3-nitro-1,5-naphthyridine-2(1H )-on (56-58); 6-bromo-4-(4-((4-fluorophenyl)(2methoxy-6-methylphenyl)methyl)piperazin-1 -yl)-1 -methyl-3-nitro-1,5-naphthyridine-2(1H) -on (59-61); tert-butyl (8-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridin-2-yl )carbamate (62); 6-amino-4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-1-methyl-3-nitro-1,5naphthyridin-2(1H)-one (63); 6-bromo-4-(4-(2-(difluoromethyl)benzyl)piperazin-1-yl)-2-oxo-1-(prop-2-yn-1yl)-1,2-dihydro-1,5- naphthyridine-3-carbonitrile (64); 6-bromo-4-(4-(2-hydroxybenzyl)piperazin-1-yl)-2oxo-1 -(prop-2-in-1 -yl)-1,2-dihydro-1,5-naphthyridin-3 -carbonitrile (65); 6-bromo-4-(4-(2-hydroxy-4methylbenzyl)piperazin-1 -yl)-2-oxo-1 -(prop-2-in-1 -yl)-1,2-dihydro-1,5 -naphthyridine-3-carbonitrile (66); 6-bromo-4-(4-(4-fluoro-2-hydroxybenzyl)piperazin-1-yl)-2-oxo-1 -(prop-2-in-1 -yl)-1,2-dihydro-1 ,5naphthyridine-3-carbonitrile (67); 6-bromo-4-(4-(4-fluoro-2-methoxybenzyl)piperazin-1-yl)-2-oxo-1-(prop- 3 044500

2-yn-1-yl)-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (68); 6-bromo-4-(4-(2-hydroxy-4,6-dimethylbenzyl)piperazin-1 -yl)-2-oxo-1 -(prop-2-in-1-yl)-1,2-dihydro -1,5-naphthyridine-3-carbonitrile (69); 6-bromo4-(4-((4-fluorophenyl)(2-hydroxyphenyl)methyl)piperazin-1 -yl)-1 -methyl-3-nitro-1,5-naphthyridin-2(1 H)-one (70 ); 6-bromo-4-(4-((2-fluoro-4-methylphenyl)(2-hydroxyphenyl)methyl)piperazin-1-yl)-1-methyl-3-nitro-1,5naphthyridine-2(1H) -on (71); 6-bromo-4-(4-((2,4-dimethylphenyl)(2-hydroxyphenyl)methyl)piperazin-1 -yl)-1 methyl-3-nitro-1,5-naphthyridin-2(1H)-one (72); 6-bromo-4-(4-((2-hydroxyphenyl)(ortho-tolyl)methyl)piperazin-1-yl)-1-methyl-3-nitro-1,5-naphthyridin-2(1H)-one ( 73); 6-bromo-4-(4-((3-fluoro-2-hydroxyphenyl)(4fluorophenyl)methyl)piperazin-1-yl)-1-methyl-3-nitro-1,5-naphthyridine-2(1H)- he (74-76); 6-bromo-4-(4-((2hydroxyphenyl) (phenyl)methyl)piperazin-1 -yl)-1 -methyl-3-nitro-1,5-naphthyridin-2(1 H)-one (77-79 ); 6bromo-4-(4-((4-fluoro-2-hydroxyphenyl)(4-fluorophenyl)methyl)piperazin-1-yl)-1-methyl-3-nitro-1,5-naphthyridine-2(III)- he (80-82); 6-bromo-4-(4-((4-fluorophenyl)(2-hydroxy-3-methylphenyl)methyl)piperazin-1-yl)1-methyl-3-nitro-1,5-naphthyridine-2(1H) -on (83-85); 6-bromo-4-(4-((4-fluorophenyl)(2-hydroxy-5-methylphenyl)methyl)piperazin-1 -yl)-1 -methyl-3-nitro-1,5-naphthyridin-2(1 N)-on (86); 6-bromo-4-(4-((4-fluorophenyl)(2-hydroxy-6-methylphenyl)methyl)piperazin-1 -yl)-1 -methyl-3-nitro-1,5-naphthyridin-2(1 N)-on (87); 5-((2-((4-(6-bromo-1-methyl-3-nitro-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)piperazin-1-yl)( 4fluorophenyl)methyl)-3-methylphenoxy)methyl)nicotinonitrile (88-90); 5-((2-((4-(6-bromo-1-methyl-3-nitro-2oxo-1,2-dihydro-1,5-naphthyridin-4-yl)piperazin-1-yl)(4- fluorophenyl)methyl)-5-fluorophenoxy)methyl)nicotinonitrile (91-93); 4-(4-((2-(allyloxy)-6-methylphenyl)(4-fluorophenyl)methyl)piperazin-1-yl)-6-bromo1-methyl-3-nitro-1,5-naphthyridine-2(1H )-on (94-96); 8-(4-((4-fluorophenyl)(2-hydroxyphenyl)methyl)piperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridin-2 -carbonitrile (97-99); 8-(4-((4fluorophenyl)(2-(prop-2-yn-1-yloxy)phenyl)methyl)piperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6- dihydro1,5-naphthyridine-2-carbonitrile (100-102); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-6-methoxy-1methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (103); 4-(4-((4-fluorophenyl)(2-hydroxyphenyl)methyl)piperazin-1 -yl)-6-methoxy-1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3 -carbonitrile (104); 6-bromo-4-(4-((4-fluorophenyl)(2-hydroxyphenyl)methyl)piperazin-1-yl)-2-oxo-1-(prop-2-yn-1-yl)-1,2dihydro -1,5-naphthyridine-3-carbonitrile (105-107); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1 -yl)-6bromo-1-(2-methoxyethyl)-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (113); 6-bromo-4-(4-((4-fluoro2-methoxyphenyl)(4-fluorophenyl)methyl)piperazin-1-yl)-1-(2-methoxyethyl)-2-oxo-1,2-dihydro-1 ,5-naphthyridine-3-carbonitrile (114-116); 6-bromo-4-(4-((4-fluorophenyl)(2-hydroxyphenyl)methyl)piperazin-1-yl)1 -(2-methoxyethyl)-2-oxo-1,2-dihydro-1,5- naphthyridine-3-carbonitrile (117); 8-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-7-nitro-6-oxo-5-(prop-2-in-1-yl)-5,6-dihydro-1 ,5-naphthyridine-2-carbonitrile (118); 8-(4-((4-fluoro-2-hydroxyphenyl)(4-fluorophenyl)methyl)piperazin-1-yl)-7-nitro-6-oxo-5-(prop-2in-1-yl)-5 ,6-dihydro-1,5-naphthyridine-2-carbonitrile (119-121); 8-(4-((4-fluoro-2-hydroxyphenyl)(4-fluorophenyl)methyl)piperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5 -naphthyridine-2-carbonitrile (122); 8-(4-((4-fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl)piperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5 -naphthyridine-2-carbonitrile (123-125); 8-(4-((4-fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl)piperazin-1-yl)-7-nitro-6-oxo-5-(prop-2-in-1-yl) -5,6-dihydro-1,5-naphthyridine-2-carbonitrile (126-128); 8-(4-((4-fluorophenyl)(2-methoxyphenyl)methyl)piperazin-1-yl)-7-nitro-6-oxo-5-(prop-2-yn-1yl)-5,6-dihydro -1,5-naphthyridine-2-carbonitrile (129-131); 8-(4-((4-fluorophenyl)(2hydroxyphenyl)methyl)piperazin-1-yl)-7-nitro-6-oxo-5-(prop-2-yn-1-yl)-5,6-dihydro -1,5-naphthyridine2-carbonitrile (132-134); 8-(4-(1-(4-fluorophenyl)ethyl)piperazin-1-yl)-7-nitro-6-oxo-5-(prop-2-in-1-yl)5,6-dihydro-1 ,5-naphthyridine-2-carbonitrile (135-137); 8-(4-(6-methoxy-2,3-dihydro-1H-inden-1yl)piperazin-1-yl)-7-nitro-6-oxo-5-(prop-2-in-1-yl) -5,6-dihydro-1,5-naphthyridine-2-carbonitrile (138); 8-{4-[1-(4-fluorophenyl)propyl]piperazin-1-yl}-7-nitro-6-oxo-5-(prop-2-yn-1-yl)-5,6-dihydro- 1,5naphthyridine-2-carbonitrile (139-141); 8-(4-(cyclopropyl(4-fluorophenyl)methyl)piperazin-1-yl)-7-nitro6-oxo-5-(prop-2-in-1-yl)-5,6-dihydro-1,5 -naphthyridine-2-carbonitrile (142-144); 8-(4-((4-fluorophenyl)(2hydroxyphenyl)methyl)piperazin-1-yl)-5-(2-methoxyethyl)-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine -2carbonitrile (145); 8-(4-(6-methoxy-2,3-dihydro-1H-inden-1 -yl)piperazin-1 -yl)-5-(2-methoxyethyl)-7nitro-6-oxo-5,6-dihydro -1,5-naphthyridine-2-carbonitrile (146); 8-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-5-(2-methoxyethyl)-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2 -carbonitrile (147); 8-(4(1-(4-fluorophenyl)ethyl)piperazin-1-yl)-5-(2-methoxyethyl)-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2carbonitrile (148-150); 5-(cyanomethyl)-8-(4-(1-(4-fluorophenyl)ethyl)piperazin-1-yl)-7-nitro-6-oxo-5,6dihydro-1,5-naphthyridine-2-carbonitrile ( 151); 8-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-5(cyanomethyl)-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridin-2-carbonitrile ( 152); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-6-bromo-1-(cyanomethyl)-2-oxo-1,2-dihydro-1,5-naphthyridin-3-carbonitrile (153); 6-bromo-1-(cyanomethyl)-4-(4-(1-(4-fluorophenyl)ethyl)piperazin-1-yl)-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile ( 154); 6-bromo-1 -(cyanomethyl)-4-(4-( 1 -(4-fluorophenyl)propyl)piperazin-1 -yl)2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (155); 6-bromo-1-(cyanomethyl)-4-(4-(6-methoxy-2,3dihydro-1H-inden-1 -yl)piperazin-1 -yl)-2-oxo-1,2-dihydro-1 ,5-naphthyridine-3-carbonitrile (156); 6bromo-1-(cyanomethyl)-4-(4-((4-fluorophenyl)(2-hydroxyphenyl)methyl)piperazin-1-yl)-2-oxo-1,2-dihydro1,5-naphthyridine-3-carbonitrile (157); 6-bromo-1-(cyclopropylmethyl)-4-(4-((4-fluorophenyl)(2-methoxyphenyl)methyl)piperazin-1 -yl)-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile (158-160); 4-(4-(bis(4- 4 044500 fluorophenyl)methyl)piperazin-1 -yl)-6-bromo-1 -(cyclopropylmethyl)-2-oxo-1,2-dihydro-1,5-naphthyridin-3 carbonitrile (161); 6-bromo-1 -(cyclopropylmethyl)-4-(4-( 1 -(4-fluorophenyl)ethyl)piperazin-1 -yl)-2-oxo1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 162-164); 4-(4-benzhydrylpiperazin-1 -yl)-3-nitro-1 -(prop2-yn-1 -yl)-1,5-naphthyridin-2(1 H)-one (165); 1-(2-(1,3-dioxan-2-yl)ethyl)-4-(4-benzhydrylpiperazin-1 -yl)3-nitro-1,5-naphthyridin-2(1 H)-one (166) ; 1 -allyl-4-(4-benzhydrylpiperazin-1 -yl)-3-nitro-1,5-naphthyridin-2(1 H)-one (176); 4-(4-benzhydrylpiperazin-1 -yl)-1-butyl-3-nitro-1,5-naphthyridin-2(1 H)-one (177); 4-(4-(4-benzhydrylpiperazin-1-yl)-3-nitro-2-oxo-1,5-naphthyridin-1(2H)-yl)butanenitrile (183); 4-(4benzhydrylpiperazin-1 -yl)-3-nitro-1 -(3,3,3-trifluoropropyl)-1,5-naphthyridin-2(1H)-one (185); 4-(4benzhydrylpiperazin-1 -yl)-1 -(4,4-difluorobut-3-en-1 -yl)-3-nitro-1,5-naphthyridin-2(1 H)-one (186); 4-(4benzhydrylpiperazin-1 -yl)-3-nitro-1 -(4-oxopentyl)-1,5-naphthyridin-2(1 H)-one (187); 4-(4-benzhydrylpiperazin-1 -yl)-1 -(3-(2-methoxyethoxy)propyl)-3-nitro-1,5-naphthyridin-2(1 H)-one (189); 4-(4-benzhydrylpiperazin-1 -yl)-1-(3-methoxypropyl)-3-nitro-1,5-naphthyridin-2(1H)-one (190); 4-(4-benzhydrylpiperazin-1 -yl)-6-bromo-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3-carbonitrile (191); 6-bromo-4(4-(4-fluoro-2-hydroxybenzyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (192) ; 6-chloro-4-(4-(4-fluoro-2-hydroxybenzyl)piperazin-1 -yl)-1 -methyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile (193); 4-(4-(4-fluoro-2-hydroxybenzyl)piperazin-1 -yl)-1,6-dimethyl-2-oxo1,2-dihydro-1,5-naphthyridine-3-carbonitrile (194); 4-(4-(1-(2-(allyloxy)-4-fluorophenyl)ethyl)piperazin-1yl)-6-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine- 3-carbonitrile (195); 4-(4-benzhydrylpiperazin1-yl)-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (196); 6-chloro-4-(4-(2-hydroxybenzyl)piperazin-1 -yl)-1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (198); 6-chloro-4-(4-(3 (ethyl(methyl)amino)benzyl)piperazin-1 -yl)-1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (199); 4-(4-(7-(allyloxy)-5-fluoro-2,3-dihydro-1 H-inden-1 -yl)piperazin-1 -yl)-6-chloro-1 -methyl-2-oxo1, 2-dihydro-1,5-naphthyridine-3-carbonitrile (200); 6-chloro-4-(4-(5-fluoro-7-hydroxy-2,3-dihydro-Shinden-1 -yl)piperazin-1 -yl)-1 -methyl-2-oxo-1,2-dihydro -1,5-naphthyridine-3-carbonitrile (201); 4-(4benzhydrylpiperazin-1 -yl)-6-ethyl-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3-carbonitrile (202); 4(4-benzhydrylpiperazin-1 -yl)-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3-carbonitrile (204); 4(4-benzhydrylpiperazin-1 -yl)-1-methyl-2-oxo-6-vinyl-1,2-dihydro-1,5-naphthyridin-3-carbonitrile (208); 6-chloro-4-(4-(cyclohexyl(phenyl)methyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (209); 4-(4-(2-aminobenzyl)piperazin-1-yl)-6-bromo-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridin-3-carbonitrile (211); K-(2-((4-(6-bromo-3-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin4-yl)piperazin-1-yl)methyl)phenyl) methanesulfonamide (212); K-(2-((4-(6-bromo-3-cyano-1-methyl-2-oxo1,2-dihydro-1,5-naphthyridin-4-yl)piperazin-1 -yl)methyl)phenyl) benzamide (213); 1 -(2-((4-(6-bromo-3 cyano-1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)piperazin-1 -yl)methyl)phenyl )-3-phenylurea (214); K-(2-((4-(6-bromo-3-cyano-1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)piperazin-1 -yl)methyl) phenyl)acetamide (215); 6-chloro-4-(4-(indolin-7-ylmethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3-carbonitrile (216); 8-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-5-methyl-6oxo-5,6-dihydro-1,5-naphthyridine-2,7-dicarbonitrile (217); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1yl)-6-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (218); 6-chloro-4-(4-((2-hydroxyphenyl)(phenyl)methyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (219); 6-chloro-4-(4-(( 1 -ethyl-1H-indol-4-yl)methyl)piperazin-1 -yl)-1 -methyl-2-oxo-1,2-dihydro-1,5- naphthyridine3-carbonitrile (220); 6-chloro-1-methyl-4-(4-(naphthalene-1 -ylmethyl)piperazin-1 -yl)-2-oxo-1,2-dihydro-1,5naphthyridin-3-carbonitrile (221); 6-chloro-4-(4-((4-fluoro-2-hydroxyphenyl)(4-fluorophenyl)methyl)piperazin-1 -yl)-1 -methyl-2-oxo-1,2-dihydro-1,5 -naphthyridine-3-carbonitrile (222); 8-(4-benzhydrylpiperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridin-2-carbonitrile (223); 3-bromo-4-(4(1 -(4-fluorophenyl)ethyl)piperazin-1 -yl)-1,6-dimethyl-1,5-naphthyridin-2(1H)-one (224); 6-bromo-4-(4-(( 1 -ethyl1H-indol-4-yl)methyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine- 3-carbonitrile (225); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1 -yl)-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (226); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1 -yl)-1 -methyl-3 -(2,2,2-trifluoroacetyl)-1,5naphthyridin-2(1 H)-one (227 ); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-1,6-dimethyl-3-(2,2,2trifluoroacetyl)-1,5-naphthyridin-2(1H)-one ( 228); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-6-bromo2-oxo-1-(prop-2-yn-1-yl)-1,2-dihydro-1,5 -naphthyridine-3-carbonitrile (229); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-3-bromo-1-methyl-1,5-naphthyridin-2(1H)-one (230); 8-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-7-cyano-K,K,5-trimethyl-6-oxo-5,6-dihydro-1,5-naphthyridin-2 -carboxamide (231); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1 -yl)-1,6-dimethyl-3-(trifluoromethyl)-1,5-naphthyridin-2(1H)-one (232); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-3-chloro-1,6-dimethyl-1,5-naphthyridin-2(1H)-one (233); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-3-fluoro-1,6-dimethyl-1,5-naphthyridin-2(1H)-one (234); 4-(4(bis(4-fluorophenyl)methyl)piperazin-1-yl)-6-bromo-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile, TFU ( 235); 3 -bromo-4-(4-(( 1 -ethyl-1 H-indol-4-yl)methyl)piperazin-1 -yl)-1 -methyl-1,5-naphthyridin-2(1 H)-one (236); 6-bromo-1-methyl-4-(4-(naphthalene-1 -ylmethyl)piperazin-1 -yl)-2-oxo-1,2-dihydro1,5-naphthyridin-3-carbonitrile (237); 4-(4-([1,1'-biphenyl]-2-ylmethyl)piperazin-1-yl)-6-bromo-1-methyl-2oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile, TFU (238); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1 yl)-3-bromo-1,6-dimethyl-1,5-naphthyridin-2(1H)-one (239); methyl 8 -(4-(bis(4-fluorophenyl)methyl)piperazine- 5 044500

1-yl)-7-cyano-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carboxylate (240); 8-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-7-cyano-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carboxylic acid ( 241); 4-[4-(diphenylmethyl)piperazin-1 -yl]-1 -methyl-3-nitro-1,2-dihydro-1,5-naphthyridin-2-one (242); 4[4-(diphenylmethyl)piperazin-1-yl]-1-ethyl-3-nitro-1,2-dihydro-1,5-naphthyridin-2-one (243); 4-[4-(diphenylmethyl)piperazin-1-yl]-1-(2-methoxyethyl)-3-nitro-1,2-dihydro-1,5-naphthyridin-2-one (244); 2-{4-[4(diphenylmethyl)piperazin-1 -yl]-3-nitro-2-oxo-1,2-dihydro-1,5-naphthyridin-1 -yl}acetonitrile (245);

ethyl 2-{4-[4-(diphenylmethyl)piperazin-1 -yl]-3-nitro-2-oxo-1,2-dihydro-1,5-naphthyridin-1 -yl} acetate (246); 4-[4-(diphenylmethyl)piperazin-1-yl]-3-nitro-1-propyl-1,2-dihydro-1,5-naphthyridin-2-one (248); 4{4-[cyclopropyl(4-fluorophenyl)methyl]piperazin-1-yl}-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridine3-carbonitrile (250); 4-{4-[bis(4-fluoro-2-methoxyphenyl)methyl]piperazin-1-yl}-1,6-dimethyl-2-oxo-1,2dihydro-1,5-naphthyridine-3-carbonitrile (251 ); 4-[4-(4-methoxybutan-2-yl)piperazin-1 -yl]-1,6-dimethyl2-oxo-1,2-dihydro-1,5-naphthyridin-3-carbonitrile (252); 4-[4-(3,4-dihydro-2H-1-benzopyran-4-yl)piperazin-1 -yl]-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile (253); 4-{4-[(4-fluorophenyl)(2-methoxypyridin-3 -yl)methyl]piperazin-1 -yl}-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile (254); 4-{4-[(4-fluorophenyl)(3-methoxypyridin-2-yl)methyl]piperazin-1-yl}-1,6dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3 -carbonitrile (255); 4-{4-[(4-fluorophenyl)(pyridin-2yl)methyl]piperazin-1-yl}-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (256); 4-{4[(2-bromo-6-hydroxyphenyl)methyl]piperazin-1-yl}-6-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine3-carbonitrile (258 ); 6-bromo-4-{4-[(2-hydroxy-6-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 259); 8-{4-[2-(4-fluorophenyl)propan-2-yl]piperazin-1-yl}-5methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridin-2- carbonitrile (260); 4-{4-[(4-tert-butyl-2-hydroxyphenyl)methyl]piperazin-1-yl}-6-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin- 3-carbonitrile (261); 4-(4-{[2-hydroxy-5-(trifluoromethoxy)phenyl]methyl}piperazin-1-yl)-1,6-dimethyl-2-oxo-1,2-dihydro1,5-naphthyridine-3-carbonitrile (262); 8-{4-[(4-bromo-2-hydroxyphenyl)methyl]piperazin-1-yl}-5-methyl-6oxo-5,6-dihydro-1,5-naphthyridine-2,7-dicarbonitrile (263) ; 6-chloro-4-{4-[(2-hydroxy-6-methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (264); 8-(4-{[2hydroxy-4-(trifluoromethoxy)phenyl]methyl}piperazin-1-yl)-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2,7-dicarbonitrile (265); 6-bromo-4-{4-[(4-chloro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 266); 6-chloro-4-{4-[(2-chloro-6-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (267); 4-{4-[(3-bromo-2hydroxyphenyl)methyl]piperazin-1-yl}-6-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 268); 8-{4-[(4-chloro-2-hydroxyphenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro-1,5naphthyridine-2,7-dicarbonitrile (269) ; 6-bromo-4-(4-{[2-hydroxy-5-(trifluoromethoxy)phenyl]methyl}piperazin-1 -yl)-1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile (270); 6-bromo-4-{4-[(2-bromo-6hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 271); 6-chloro-4-{4-[(2-chlorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3carbonitrile (272); 6-chloro-4-{4-[(2-hydroxy-4-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridine-3-carbonitrile ( 273); 6-chloro-4-{4-[(3-fluoro-2-methoxyphenyl)methyl]piperazin1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 274); 4-{4-[(5-bromo-2-hydroxyphenyl)methyl]piperazin-1-yl}-6-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (275); 4-{4-[(4-bromo-2-hydroxyphenyl)methyl]piperazin-1-yl}-6-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (276); 4-{4-[(5-bromo-2-hydroxyphenyl)methyl]piperazin-1-yl}-1,6-dimethyl-2-oxo1,2-dihydro-1,5-naphthyridine-3-carbonitrile (277) ; 6-bromo-4-{4-[(3-chloro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (278); 6-chloro-4-{4-[(3-fluoro-4methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 279); 6-chloro-4-{4-[(2-fluoro-6-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (280); 6-chloro-4-{4-[(2-fluoro-3-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 281); 6-bromo-4-{4-[(5-bromo-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (282); 6-chloro-4-{4-[(2-hydroxy5-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 283); tert-butyl N-(2-{ [4-(6-bromo-3-cyano-1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)piperazin-1 yl] methyl}phenyl)carbamate (284); 6-chloro-1-methyl-2-oxo-4-(4-{[2-(trifluoromethoxy)phenyl]methyl}piperazin-1-yl)-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (285); 4-{4-[(3-bromo-2-hydroxyphenyl)methyl]piperazin-1-yl}-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 286); 6-chloro-4-{4-[(2chloro-4-fluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 287); 6-chloro-4-{4-[(4-chloro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile ( 288); 6-chloro-4-{4-[(3-fluoro-4-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 289); 6-chloro-4-{4-[(3-fluoro-5-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (290); 6-chloro-4-{4-[(3,5-difluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 291); 6chloro-4-{4-[(2-hydroxy-6-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 292); 6-chloro-4-{4-[(3-chloro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo- 6 044500

1,2-dihydro-1,5-naphthyridine-3-carbonitrile (293); 6-bromo-4-{4-[(2-fluoro-6-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (294); 6-bromo-4-{4-[(2-hydroxy4-methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 295); 4-{4-[(5-chloro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 296); 6-bromo-4-{4-[(2-methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridine-3-carbonitrile (297); 6-chloro-4-{4-[(3,5-difluoro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine- 3-carbonitrile (298); 6-bromo-4-{4-[(2,3dihydro-III-indol-7-yl)methyl] piperazin-1 -yl} -1 -methyl-2-oxo-1,2-dihydro-1,5- naphthyridine-3-carbonitrile (299); 6-bromo-4-{4-[(2-chlorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile (300); 6-chloro-4-{4-[(2-hydroxy-4-methoxyphenyl)methyl]piperazin-1-yl}-1methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 301); 6-chloro-4-{4-[(3-chlorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (302) ; 6-chloro-4-{4-[(III-indol-7yl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 303); 6-bromo-4{4-[(3-fluoro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3carbonitrile (304 ); 6-chloro-4-{4-[(2-hydroxy-5-methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 305); 6-chloro-4-{4-[(3-fluoro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (306); 6-chloro-4-{4-[(3,5-dichloro-2hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (307); 6-chloro-4-{4-[(4-fluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3carbonitrile (308); 6-bromo-4-{4-[(3-tert-butyl-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo1,2-dihydro-1,5-naphthyridin-3- carbonitrile (309); 6-chloro-4-{4-[(2,4-dichlorophenyl)methyl]piperazin-1yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (310) ; 6-chloro-4-(4-{[2-hydroxy-4(trifluoromethyl)phenyl]methyl} piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin- 3-carbonitrile (311); 6-chloro-4-{4-[(5-fluoro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridine-3-carbonitrile ( 312); 6-chloro-4-(4-{[3-fluoro-5-(trifluoromethyl)phenyl]methyl}piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile (313); 6-bromo-4-{4-[(2hydroxy-6-methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 314); 6-chloro-4-{4-[1-(4-fluorophenyl)-2-methylpropyl]piperazin-1-yl}-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridine-3-carbonitrile (315-316); 8-{4-[(2-hydroxyphenyl)methyl]piperazin-1-yl}-5methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2,7-dicarbonitrile (317); 6-chloro-4-{4-[(2,5-difluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 318); 6-chloro-4-{4[(3,4-difluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (319 ); 6-bromo-4-{4-[(3,5-difluoro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro1,5-naphthyridin-3- carbonitrile (320); 6-chloro-4-{4-[(3-fluorophenyl)methyl]piperazin-1-yl}-1-methyl-2oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (321); 6-chloro-4-{4-[(1H-indazol-7-yl)methyl]piperazin1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3-carbonitrile ( 322); 6-chloro-4-{4-[(4-chloro-3-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (323); 6chloro-4-{4-[(3-chloro-5-fluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3carbonitrile (324) ; 8-[4-(diphenylmethyl)piperazin-1-yl]-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine2,7-dicarbonitrile (325); 6-chloro-4-{4-[(4-chloro-3-fluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridine-3-carbonitrile ( 326); 6-chloro-4-{4-[(3-methoxyphenyl)methyl]piperazin-1-yl}-1methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (327); 6-chloro-4-{4-[(3-hydroxy-4-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (328); 4-{4-[(3fluoro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3carbonitrile (329); 6-chloro-4-{4-[(2,3-difluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro1,5-naphthyridine-3-carbonitrile (330) ; 6-chloro-4-{4-[(2-methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (331); 8-{4-[(4-chloro-3-hydroxyphenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2,7-dicarbonitrile ( 332); 6-bromo-4-{4-[(Shindazol-7-yl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3-carbonitrile ( 333); 4-{4-[(1 -ethyl-1 H-indol-4-yl)methyl]piperazin-1 -yl}-1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3carbonitrile (334); 6-chloro-4-{4-[(2-fluoro-6-methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridine-3-carbonitrile ( 335); 6-chloro-4-{4-[(2-hydroxynaphthalene-1-yl)methyl]piperazin1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3-carbonitrile ( 336); 4-{4-[(2-hydroxyphenyl)methyl]piperazin-1-yl}-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (337); 6-chloro-4-{4[(3-fluoro-5-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (338); 6-chloro-4-{4-[(2-fluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile (339); 6-chloro-4-{4-[(5-cyano-2-fluorophenyl)methyl]piperazin-1-yl}-1-methyl-2oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 340); 6-chloro-4-{4-[(4-methoxyphenyl)methyl]piperazin1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (341); 8-{4-[1-(4-fluorophenyl)cyclopropyl]piperazin-1-yl}-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile ( 342); 4-(4benzylpiperazin-1-yl)-6-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3-carbonitrile (343); 6-bromo4-{4-[(2-fluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile

- 7 044500 (344); 6-chloro-4-{4-[(3-chloro-5-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile ( 345); 6-chloro-1-methyl-4-{4-[(2-methylphenyl)methyl]piperazin-1-yl}-2-oxo1,2-dihydro-1,5-naphthyridine-3-carbonitrile (346); 4-{4-[bis(4-fluorophenyl)methyl]piperazin-1-yl}-1methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (347); 6-chloro-4-{4-[(4-fluoro-3-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (348); 4-{4-[(2hydroxyphenyl)(phenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (349); 6-chloro-4-{4-[(2,4-difluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile (350) ; 6-chloro-4-{4-[(3-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (351); 4-{4-[(2-hydroxy-3-methoxyphenyl)methyl]piperazin-1-yl}-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 352); 6-chloro-4-{4-[(3hydroxy-4-methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 353); 6-chloro-4-{4-[(3-chloro-4-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridine-3-carbonitrile ( 354); 6-chloro-4-(4-{[4-hydroxy-3-(trifluoromethyl)phenyl]methyl} piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile (355); 6-chloro-4-{4-[(4hydroxy-3-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 356); 6-chloro-4-{4-[(3-fluoro-4-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridine-3-carbonitrile ( 357); 6-chloro-4-[4-(diphenylmethyl)piperazin-1-yl]-1-methyl-2oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (358); 4-{4-[(1H-1,3-benzodiazol-7-yl)methyl]piperazin-1-yl}-6-chloro-1-methyl-2-oxo-1,2-dihydro-1,5- naphthyridine-3-carbonitrile (359); 4-{4-[(1H-1,3benzodiazol-7-yl)methyl]piperazin-1 -yl}-6-bromo-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin- 3-carbonitrile (360); 6-chloro-4-{4-[(4-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile (362); 8-{4-[(3-fluoro-4-hydroxyphenyl)methyl]piperazin-1-yl}-5-methyl-6oxo-5,6-dihydro-1,5-naphthyridine-2,7-dicarbonitrile (363) ; 6-chloro-4-{4-[(2-fluoro-5-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (3 64); 6-chloro-4-{4-[(2chloro-6-methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 365); 6-bromo-4-{4-[(3,5-dichloro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridin-3- carbonitrile (366); 6-bromo-4-{4-[(5-chloro-2-hydroxyphenyl)methyl]piperazin1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 367); 6-bromo-4-{4-[(2-hydroxy-3methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 368); 6-chloro-4-(4-{[2-hydroxy-5-(trifluoromethoxy)phenyl]methyl}piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydro1,5-naphthyridin-3 -carbonitrile (369); 6-chloro-4-{4-[(5-chloro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 370); 6-chloro-4-{4-[(2-hydroxy-3-methoxyphenyl)methyl]piperazin-1 -yl} -1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3 - carbonitrile (371); 6-bromo4-{4-[(2-hydroxy-4-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3carbonitrile (372) ; 6-bromo-4-{4-[(1 H-indol-7-yl)methyl]piperazin-1 -yl}-1 -methyl-2-oxo-1,2-dihydro1,5-naphthyridine-3-carbonitrile (373); 6-bromo-1-methyl-2-oxo-4-{4-[(2-oxo-2,3-dihydro-1H-indol-7yl)methyl]piperazin-1-yl}-1,2-dihydro- 1,5-naphthyridine-3-carbonitrile (374); 6-chloro-4-(4-{[3-fluoro-4(trifluoromethyl)phenyl]methyl} piperazin-1-yl)-1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridin- 3-carbonitrile (375); 6-chloro-1-methyl-2-oxo-4-{4-[(2-oxo-2,3-dihydro-1H-indol-7-yl)methyl]piperazin-1-yl}-1,2dihydro- 1,5-naphthyridine-3-carbonitrile (376); 6-chloro-4-{4-[1-(4-fluorophenyl)-3,3-dimethylbutyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile (377-378); 6-chloro-4-{4-[(4-cyano-2-fluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (379); 6-bromo-4-{4-[(3-bromo-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile ( 380); 6-chloro-4-(4-{[2-hydroxy-4-(trifluoromethoxy)phenyl]methyl}piperazin1-yl)-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3 -carbonitrile (381); 6-chloro-4-{4-[cyclobutyl(4fluorophenyl)methyl]piperazin-1 -yl}-1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (382383); 4-{4-[(3-tert-butyl-2-hydroxyphenyl)methyl]piperazin-1-yl}-6-chloro-1-methyl-2-oxo-1,2-dihydro1,5-naphthyridin-3- carbonitrile (384); 6-chloro-1-methyl-4-{4-[(3-methylphenyl)methyl]piperazin-1-yl}-2oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (385); 6-chloro-1-methyl-4-{4-[(4-methylphenyl)methyl]piperazin-1-yl}-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (386) ; 6-chloro-4-{4-[1-(4-fluorophenyl)ethyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 387-388); 4-{4[(3-chloro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3carbonitrile (389); 6-bromo-4-{4-[(2-chloro-6-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridine-3-carbonitrile ( 390); 6-chloro-4-{4-[(2-hydroxy-3-methylphenyl)methyl]piperazin1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 391); 8-{4-[(4-fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridin-2 -carbonitrile (392); 4-{4-[bis(4-chlorophenyl)methyl]piperazin-1-yl}-6-bromo-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (393 ); 8-{4-[bis(4-chlorophenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro1,5-naphthyridine-2,7-dicarbonitrile (394); 4-{4-[bis(4-chlorophenyl)methyl]piperazin-1-yl}-6-methoxy-1methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (395); 4-{4-[(4-fluorophenyl)(2-methoxypyridin3-yl)methyl]piperazin-1-yl}-6-methoxy-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile (396); 4-{4-[(4-fluorophenyl)(3-methoxypyridin-2-yl)methyl]piperazin-1-yl}-6-methoxy-1-methyl-2-oxo

- 8 044500

1,2-dihydro-1,5-naphthyridine-3-carbonitrile (397); 4-{4-[1-(4-fluorophenyl)propyl]piperazin-1-yl}-1,6dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (398); 8-{4-[(S)-(4-chlorophenyl)(phenyl)methyl]piperazin-1-yl}-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine -2-carbonitrile (399); 8-{4-[(4fluorophenyl)(pyridin-2-yl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridin-2,7dicarbonitrile (400 ); 4-{4-[(4-fluoro-2-methoxyphenyl)(pyrimidin-2-yl)methyl]piperazin-1-yl}-6-methoxy-1 -methyl-2-oxo-1,2-dihydro-1 ,5-naphthyridine-3-carbonitrile (401); 4-{4-[bis(4-fluoro-2-methoxyphenyl)methyl]piperazin-1 -yl}-6-methoxy-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3 -carbonitrile (402); 4-{4-[(4-fluorophenyl)(pyridin-2-yl)methyl]piperazin-1 -yl}-6-methoxy-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridin-3 -carbonitrile (403); 5-methyl-8-{4-[(naphthalene-1-yl)methyl]piperazin-1-yl}-6-oxo-5,6dihydro-1,5-naphthyridin-2,7-dicarbonitrile (404); 8-{4-[(4-fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridin-2 ,7-dicarbonitrile (405-407); 5-methyl8-{4-[(4-methylphenyl)(phenyl)methyl]piperazin-1-yl}-6-oxo-5,6-dihydro-1,5-naphthyridine-2,7-dicarbonitrile (408); 8-{4-[bis(4-fluoro-2-methoxyphenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro-1,5naphthyridine-2,7-dicarbonitrile (409 ); 8-{4-[(4-fluorophenyl)(3-methoxypyridin-2-yl)methyl]piperazin-1yl}-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridin-2,7 -dicarbonitrile (410); 8-{4-[(4-fluorophenyl)(phenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2,7-dicarbonitrile (411 ); 4-{4[(4-fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl]piperazin-1-yl}-6-methoxy-1-methyl-2-oxo-1,2-dihydro1,5-naphthyridine- 3-carbonitrile (412); 4-{4-[(4-fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl]piperazin-1yl}-6-methoxy-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile (413); 8-{4-[(S)-(4-chlorophenyl)(phenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2,7 -dicarbonitrile (414); 5-methyl-8-{4-[(naphthalene-1-yl)methyl]piperazin-1-yl}-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridin-2-carbonitrile ( 415); 8-{4-[(4-fluorophenyl)(2-methoxy-4-methylphenyl)methyl]piperazin-1-yl}-5-methyl7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine -2-carbonitrile (416); 6-chloro-4-{4-[(4-chlorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (417) ; 8-{4-[(2-hydroxyphenyl)(phenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2,7-dicarbonitrile (418 ); 6-chloro-4-{4-[(2-chloro-6-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (419); 6-chloro-4-{4-[(2-chloro-6-fluorophenyl)methyl]piperazin-1-yl}-1-methyl-2oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 420); 6-chloro-4-{4-[1-(4-fluorophenyl)cyclopropyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 421); 6-chloro-4-{4[(2,6-difluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (422 ); 6-chloro-4-{4-[(2-fluoro-4-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (423); 6-chloro-4-{4-[(4-cyano-2-methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 424); 6-chloro-4-{4-[2-(4-fluorophenyl)propan-2-yl]piperazin-1 -yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin- 3-carbonitrile (425); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-1-methyl-1,5-naphthyridin-2(1H)-one (426); 4-{4-[bis(4-fluorophenyl)methyl]piperazin-1 -yl}-1,6-dimethyl-1,2-dihydro-1,5-naphthyridin-2-one (427); 6-bromo-4-{4-[(S)-(4-chlorophenyl)(phenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile (428); 6-bromo-4{4-[(4-chlorophenyl)(phenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 429); 6-bromo-4-{4-[(4-fluorophenyl)(phenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro1,5-naphthyridine-3-carbonitrile (430 ); 6-bromo-1-methyl-4-{4-[(4-methylphenyl)(phenyl)methyl]piperazin-1yl}-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (431 ); or 8-{4-[bis(4-fluorophenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (432).

Other specific embodiments of the present invention may be proposed without departing from the spirit or essential features of the invention. This invention includes all combinations of aspects and/or embodiments of the invention set forth in this specification. It is intended that any and all embodiments of the present invention may be combined with any other embodiment or embodiments of the invention when further embodiments of the invention are described. It is also intended that each individual element of the embodiments of the present invention should be combined with any and all other elements of any embodiment of the invention when describing a further embodiment of the invention.

Definitions.

The features and advantages of the present invention will become more apparent to those of ordinary skill in the art from the following detailed description of the invention. It should be appreciated that certain features of the present invention, which, for clarity, are described herein in the context of individual embodiments of the invention, may also be combined in a single embodiment of the invention. Conversely, several features of the present invention, which, for brevity, are described in the context of one embodiment of the invention, can also be combined into subcombinations of features of the invention. It is intended that the exemplary or preferred embodiments described herein are illustrative and not limiting of the scope of the present invention.

- 9 044500

Unless clearly indicated otherwise, as used herein, the singular form may also include the plural form. For example, singular forms of nouns can refer to either singular nouns or singular or plural nouns.

As used herein, the phrase compounds and/or salts of these compounds refers to at least one compound, at least one salt of these compounds, or a combination thereof. For example, the phrase a compound of formula (I) and/or a salt thereof includes a compound of formula (I), two compounds of formula (I), a salt of a compound of formula (I), a compound of formula (I), and one or more salts of a compound of formula ( I) and two or more salts of the compound of formula (I).

Unless otherwise stated, any atom with unsaturated valences is intended to have hydrogen atoms in sufficient quantities to saturate those valences.

The definitions set forth in this specification supersede the definitions set forth in any patent, patent application, and/or patent application publication incorporated herein by reference.

The following are definitions of various terms used to describe the present invention. These definitions apply to the specified terms wherever these terms appear in the description of the invention (unless in specific cases the definitions of the terms contain other restrictions) alone or as part of a more common group.

The groups and substituents found herein that provide stable moieties and compounds can be selected by one skilled in the art.

According to the conventions used in the art, is used in this description in structural formulas to indicate a bond that represents the site of attachment of a moiety or substituent to the core or backbone structure.

The terms halogen and halogen as used herein refer to F, C1, Br and I.

The term cyano refers to the -CN group.

The term amino refers to the -NH2 group.

The term oxo refers to the =O group.

The term alkyl as used herein refers to saturated, branched or straight chain aliphatic hydrocarbon groups containing, for example, 1 to 12 carbon atoms, 1 to 6 carbon atoms, and 1 to 4 carbon atoms. Examples of alkyl groups include methyl (Me), ethyl (Et), propyl (such as n-propyl and iso-propyl), butyl (such as n-butyl, iso-butyl, sec-butyl and tert-butyl) and pentyl ( for example, n-pentyl, isopentyl, neopentyl), n-hexyl, 2-methylpentyl, 2-ethylbutyl, 3-methylpentyl and 4-methylpentyl. When the symbol C is followed by numbers in subscript format, the subscript specifies more specifically the number of carbon atoms that a given group can contain. For example, C1-4 alkyl means straight and branched chain alkyl groups containing from one to four carbon atoms.

As used herein, the term fluoroalkyl is intended to include branched or straight chain saturated aliphatic hydrocarbon groups having one or more fluorine atoms as substituents. For example, C1.4 fluoroalkyl is intended to include C1, C2, _C3 and C4 alkyl groups having one or more fluorine atoms as substituents.

Typical examples of fluoroalkyl groups include, but are not limited to -CF3 and -CH2CF3.

The term cyanoalkyl includes saturated branched or straight chain alkyl groups substituted by one or more cyano groups. For example, cyanoalkyl includes -CH2CN, -CH2CH2CN and C1-4 cyanoalkyl.

The term aminoalkyl includes saturated branched or straight chain alkyl groups substituted by one or more amino groups. For example, aminoalkyl includes -CH2NH2, -CH2CH2NH2 and C1_ ₄ aminoalkyl.

The term hydroxyalkyl includes saturated branched or straight chain alkyl groups substituted by one or more hydroxyl groups. For example, hydroxyalkyl includes -CH2OH, -CH2CH2OH and C1_ ₄ hydroxyalkyl.

The term alkenyl refers to a straight or branched chain hydrocarbon radical containing from 2 to 12 carbon atoms and at least one carbon-carbon double bond. Typical examples of such groups include ethenyl or allyl. For example, C2-6 alkenyl means straight and branched chain alkenyl groups containing from two to six carbon atoms.

The term alkynyl refers to a straight or branched chain hydrocarbon radical containing from 2 to 12 carbon atoms and at least one carbon-carbon triple bond. Typical examples of such groups include ethynyl. For example, C ₂ _ ₆ alkynyl means straight and branched chain alkynyl groups containing from two to six carbon atoms.

The term cycloalkyl as used herein refers to a group derived from a non-aromatic monocyclic or polycyclic hydrocarbon molecule by the removal of one hydrogen atom from a saturated ring carbon. Typical examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclopentyl and cyclohexyl. When the symbol C is followed by numbers in subscript format, the subscript specifies more specifically the number of carbon atoms that a given cycloalkyl group may contain. For example,

C ₃ - ₆ cycloalkyl means cycloalkyl groups containing from three to six carbon atoms.

The term alkoxy as used herein refers to an alkyl group connected to the main molecular moiety via an oxygen atom, for example, a methoxy group (-OCH3). For example, C1_ ₃ alkoxy means alkoxy groups containing from one to three carbon atoms.

The terms fluoroalkoxy and -O(fluoroalkyl) refer to a fluoroalkyl group, as defined above, attached via an oxygen bridge (-O-). For example, _C1-4 fluoroalkoxy is intended to include C1, C2, _C3 and _C4 fluoroalkoxy groups.

The term alcalenyl refers to a saturated carbon chain containing two points of attachment to the core or main chain structure. The alkalenyl group has the following structure: -(CH ₂ ) _n -, where n is an integer of 1 or more. Examples _of alkalenyl bridges include -CH2CH2-, -CH2CH2CH2- and -( _CH2 ) _{2_4-} .

As used herein, the phrase pharmaceutically acceptable refers to compounds, substances, compositions and/or dosage forms that have been medically assessed to be suitable for use in contact with human and animal tissue and do not cause excessive toxicity, irritation, allergic reaction or other problems or complications, and correspond to a reasonable benefit/risk ratio.

The compounds of formula (I) can form salts, which are also included within the scope of the present invention. Unless otherwise indicated, reference to a compound of the present invention is intended to include reference to one or more salts of said compound. The term salt(s) refers to acidic and/or basic salts formed with inorganic and/or organic acids and bases. In addition, the term salt(s) may include zwitterions (internal salts), for example when the compound of formula (I) contains both a basic moiety, such as an amine or a pyridine or imidazole ring, and an acidic moiety, such as a carboxylic acid. Preferred salts are pharmaceutically acceptable (ie, non-toxic, physiologically acceptable) salts, such as, for example, acceptable metal salts and amine salts, in which the cation does not significantly affect the toxicity or biological activity of the salt. However, other salts may also be used, for example, in isolation or purification steps in the process of preparing a compound, and accordingly such salts are included within the scope of the present invention. Salts of compounds of formula (I) can be obtained, for example, by reacting a compound of formula (I) with an acid or base, taken, for example, in an equivalent amount, in a medium where the salt precipitates, or in an aqueous medium, followed by lyophilization .

Typical acid addition salts include acetates (such as acetates formed with acetic acid or trihaloacetic acid, such as trifluoroacetic acid), adipates, alginates, ascorbates, aspartates, benzoates, benzenesulfonates, bisulfates, borates, butyrates, citrates, camphorates, camphorsulfonates, cyclopentane propionates, digluconates, dodecyl sulfates, ethanesulfonates, fumarates, glucoheptanoates, glycerophosphates, hemisulfates, heptanoates, hexanoates, hydrochlorides (formed with hydrochloric acid), hydrobromides (formed with hydrogen bromide), hydroiodides, maleates (formed with maleic acid), 2-hydroxyethanesulfonates, lactates, methanesulfonates (formed with methanesulfonic acid), 2-naphthalene sulfonates, nicotinates, nitrates, oxalates, pectinates, persulfates, 3-phenylpropionates, phosphates, picrates, pivalates, propionates, salicylates, succinates, sulfates (such as sulfates formed with sulfuric acid), sulfonates (such as the sulfonates mentioned herein), tartrates, thiocyanates, toluenesulfonates such as tosylates, undecanoates and the like.

Typical base salts include ammonium salts, alkali metal salts such as sodium, lithium and potassium salts; alkaline earth metal salts such as calcium and magnesium salts; barium, zinc and aluminum salts; salts formed with organic bases (for example organic amines) such as trialkylamines, for example triethylamine, procaine, dibenzylamine, N-benzyl-β-phenethylamine, 1-ephenamine, N,N'-dibenzylethylene-diamine, dehydroabiethylamine, N-ethylpiperidine, benzylamine, dicyclohexylamine or similar pharmaceutically acceptable amines, and salts formed with amino acids such as arginine, lysine and the like. Basic nitrogen-containing groups can be quaternized with agents such as lower alkyl halides (for example, methyl, ethyl, propyl and butyl chlorides, bromides and iodides), dialkyl sulfates (for example, dimethyl, diethyl, dibutyl and diamyl sulfates), long-chain halides (for example, decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides), aralkyl halides (for example, benzyl and phenethyl bromides) and others. Preferred salts include monohydrochloride, hydrogen sulfate, methanesulfonate, phosphate or nitrate salts.

The compounds of formula (I) may be presented as amorphous solids or crystalline solids. To obtain compounds of formula (I) in solid form, you can

- 11 044500 lyophilization be used.

It should also be understood that solvates (eg, hydrates) of the compounds of Formula (I) are also included within the scope of the present invention. The term solvate means the physical association of a compound of formula (I) with one or more solvent molecules, organic or inorganic. This physical association involves the formation of hydrogen bonds. In some cases, a solvate may be isolated, for example when one or more solvent molecules are incorporated into the crystal lattice of the crystalline solid. The term solvate includes both liquid phase solvates and crystal solvates. Typical solvates include hydrates, ethanolates, methanolates, isopropanolates, acetonitrile solvates, and ethyl acetate solvates. Solvation techniques are well known in the art.

Various forms of prodrugs are well known in the art and are described in the following publications:

a) The Practice of Medicinal Chemistry, Camille G. Wermuth et al., Ch 31, (Academic Press, 1996);

b) Design of Products, edited by H. Bundgaard, (Elsevier, 1985);

c) A Textbook of Drug Design and Development, P. Krogsgaard-Larson and H. Bundgaard, eds. Ch 5, pgs 113 - 191 (Harwood Academic Publishers, 1991); And

d) Hydrolysis in Drug and Drug Metabolism, Bernard Testa and Joachim M. Mayer, (Wiley-VCH, 2003).

In addition, the compounds of formula (I), after their preparation, can be isolated and purified to obtain a composition containing 99% (by weight) or more of the compound of formula (I) (substantially pure compound), which is then used as is or used for obtaining the drug in the manner described in this application. Such essentially pure compounds of formula (I) are also intended to be included within the scope of the present invention.

The phrases stable compound and stable structure refer to a compound that is stable enough to withstand isolation from a reaction mixture to an acceptable degree of purity and the technology for preparing a preparation containing the compound as an effective therapeutic agent. The present invention includes stable compounds.

The phrase a therapeutically effective amount is intended to include an amount of a single compound of the present invention, or an amount of a combination of compounds of the present invention, or an amount of a compound of the present invention in combination with other active ingredients that is effective in inhibiting DGKa and/or DGKZ or showing effectiveness in treating or preventing viral infections and proliferative disorders such as cancer.

As used herein, the terms treat or treat refer to the treatment of a disease state in a mammal, particularly a human, and include: a) preventing the occurrence of the disease state in the mammal, particularly when such mammal is predisposed to developing the disease condition but the disease state has not yet developed. diagnosed as present; b) inhibition of the disease state, that is, stopping its development; and/or c) alleviation of the disease state, that is, regression of the disease state.

The compounds of the present invention are intended to include all isotopes of the atoms present in the compounds. Isotopes include atoms that have the same atomic number but different mass numbers. As a typical non-limiting example, isotopes of hydrogen include deuterium (D) and tritium (T). Isotopes of carbon include ¹³ C and ¹⁴ C. The isotopically labeled compounds of the present invention can generally be prepared using standard techniques known to those skilled in the art, or by techniques similar to those described herein using a suitable labeled isotope of the reagent instead of the unlabeled reagent used otherwise.

The compounds of formula (I), and/or pharmaceutically acceptable salts of these compounds, can be administered by any route suitable for treating the condition, the choice of route of administration may depend on the need for targeted therapy or the amount of the compound of formula (I) to be delivered .

This invention also includes a class of pharmaceutical compositions containing a compound of formula (I), and/or pharmaceutically acceptable salts thereof, and one or more non-toxic, pharmaceutically acceptable carrier and/or diluent and/or adjuvant (collectively herein referred to as a carrier) and, if desired, other active ingredients. The compounds of formula (I) may be administered by any suitable route, preferably in the form of a pharmaceutical composition adapted for such route and in a dose effective for the intended treatment. The compounds and compositions of the present invention can be administered, for example, orally, transmucosally or parenterally, including intravascularly, intravenously, intraperitoneally, subcutaneously, intramuscularly and intrasternally, using unit dosage forms containing standard pharmaceutically acceptable carriers, adjuvants and fillers. For example, the pharmaceutical carrier may contain a mixture of mannitol or lactose and microcrystalline cellulose. The mixture may contain additional components such as a lubricant such as magnesium stearate and a disintegrant such as crospovidone. The mixture containing the carrier can be used to fill gelatin capsules or can be compressed into tablet form. The pharmaceutical composition can be administered, for example, in the form of an oral dosage form or by infusion.

The pharmaceutical composition for oral administration may be in the form of, for example, a tablet, capsule, liquid capsule, suspension or liquid. Preferably, the pharmaceutical composition is presented in unit dosage form containing a specified amount of the active ingredient. For example, the pharmaceutical composition may be in the form of a tablet or capsule containing an amount of active ingredient ranging from about 0.1 to 1000 mg, preferably from about 0.25 to 250 mg, and more preferably from about 0. 5 to 100 mg. The appropriate daily dosage for a human or other mammal may vary widely depending on the patient's condition and other factors, but can be determined using standard techniques.

Any pharmaceutical composition of the present invention may be delivered, for example, orally through any acceptable and suitable oral preparations. Typical oral preparations include, for example, tablets, lozenges, lozenges, aqueous and oily suspensions, dispersible powders or granules, emulsions, hard and soft capsules, liquid capsules, syrups and elixirs. Pharmaceutical compositions intended for oral administration can be prepared using any techniques known in the art of preparing pharmaceutical compositions intended for oral administration. To obtain pharmaceutical preparations that are palatable and palatable, at least one agent selected from sweetening agents, flavoring agents, coloring agents, anti-mucosal irritants, antioxidants and preservatives may be included in the pharmaceutical composition of the present invention.

A tablet can be prepared, for example, by mixing at least one compound of formula (I), and/or at least one pharmaceutically acceptable salt of the compound, with at least one non-toxic pharmaceutically acceptable excipient suitable for the manufacture of tablets. Typical excipients include, for example, inert diluents such as, for example, calcium carbonate, sodium carbonate, lactose, calcium phosphate and sodium phosphate; granulating and disintegrating agents such as, for example, microcrystalline cellulose, croscarmellose sodium, corn starch and alginic acid; binding agents such as, for example, starch, gelatin, polyvinylpyrrolidone and gum arabic; and lubricants such as, for example, magnesium stearate, stearic acid and talc, but are not limited to. In addition, the tablet may be uncoated or may be coated using known techniques to mask the unpleasant taste of an unpalatable drug or to retard the dissolution and absorption of the active ingredient in the gastrointestinal tract and thereby prolong the action of the active ingredient. Typical water-soluble taste masking agents include, but are not limited to, hydroxypropyl methylcellulose and hydroxypropylcellulose. Typical retardants include, but are not limited to, ethylcellulose and cellulose acetate butyrate.

Hard gelatin capsules can be prepared, for example, by mixing at least one compound of formula (I), and/or at least one salt of said compound, with at least one inert solid diluent, such as, for example, calcium carbonate, phosphate calcium and kaolin.

Soft gelatin capsules can be prepared, for example, by mixing at least one compound of formula (I), and/or at least one pharmaceutically acceptable salt thereof, with at least one water-soluble carrier, such as, for example, polyethylene glycol, and at least one oily medium, such as, for example, peanut oil, petroleum jelly and olive oil.

An aqueous suspension can be prepared, for example, by mixing at least one compound of formula (I), and/or at least one pharmaceutically acceptable salt of said compound, with at least one excipient suitable for preparing an aqueous suspension. Typical excipients suitable for making an aqueous suspension include, but are not limited to, suspending agents such as, for example, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, alginic acid, polyvinylpyrrolidone, gum tragacanth and gum arabic; dispersing or wetting agents such as, for example, natural phosphatides, eg lecithin; condensation products of alkylene oxide with fatty acids, such as, for example, polyoxyethylene stearate; condensation products of ethylene oxide with long-chain aliphatic alcohols, such as, for example, heptadecaethylene-oxycetanol; condensation products of ethylene oxide with partial esters formed by fatty acids and hexitol, such as, for example, polyoxyethylene sorbitol monooleate; and condensation products of ethylene oxide with

- 13 044500 partial esters formed by fatty acids and hexitol anhydrides, such as, for example, polyethylene sorbitan monooleate. The aqueous suspension may also contain at least one preservative, such as, for example, para-hydroxybenzoic acid ethyl n-propyl ester; at least one coloring agent; at least one flavoring agent and/or at least one sweetening agent, including, but not limited to, for example, sucrose, saccharin and aspartame.

Oily suspensions can be prepared, for example, by suspending at least one compound of formula (I), and/or at least one pharmaceutically acceptable salt of said compound, in a vegetable oil, such as, for example, peanut oil, olive oil, sesame oil and coconut oil, or in mineral oil such as petrolatum. The oil suspension may also contain at least one thickening agent such as, for example, beeswax, paraffin wax and cetyl alcohol. To obtain a palatable oil suspension, at least one of the sweetening agents and/or at least one flavoring agent may be added to the oil suspension. The oil suspension may further contain at least one preservative, including, but not limited to, an antioxidant such as, for example, butylated hydroxyanisole and alpha-tocopherol.

Dispersible powders and granules can be prepared, for example, by mixing at least one compound of formula (I), and/or at least one pharmaceutically acceptable salt of said compound, with at least one dispersing and/or wetting agent, at least one suspending agent and/or at least one preservative. Suitable dispersing agents, wetting agents and suspending agents may be selected from those already described above. Typical preservatives include, for example, but are not limited to antioxidants such as ascorbic acid. In addition, dispersible powders and granules may also contain at least one excipient, including, but not limited to, for example, sweetening agents, flavoring agents and coloring agents.

An emulsion containing at least one compound of formula (I) and/or at least one pharmaceutically acceptable salt of said compound can be prepared, for example, as an oil-in-water emulsion.

The oil phase of emulsions containing compounds of formula (I) can be formulated from known ingredients using known techniques. The oil phase can be, without limitation, for example, vegetable oil such as, for example, olive oil and peanut oil; mineral oil such as, for example, petroleum jelly; and mixtures of these oils. The oil phase may contain only an emulsifier, the oil phase may also contain a mixture of at least one emulsifier with a fat or oil component or both a fat and oil component. Suitable emulsifying agents include, but are not limited to, natural phosphatides, such as soybean lecithin; esters or partial esters formed by fatty acids and hexitol anhydrides, such as, for example, sorbitan monooleate; and condensation products of partial esters with ethylene oxide, such as, for example, polyoxyethylene sorbitan monooleate. In a preferred embodiment, the hydrophilic emulsifier is included along with a lipophilic emulsifier which acts as a stabilizer. In addition, in a preferred embodiment, both an oil and a fat component are included. The emulsifier(s), together with or without stabilizer(s), constitute the so-called emulsifying wax, and this wax, together with the oil and fat component, constitute the so-called emulsifying ointment base, which forms the oil dispersed phase of the preparations in the form of a cream. The emulsion may also contain a sweetening agent, flavoring agent, preservative and/or antioxidant. Emulsifiers and emulsion stabilizers suitable for use in the formulation of the present invention include Tween 60, Span 80, cetostearyl alcohol, myristyl alcohol, glycerol monostearate, sodium lauryl sulfate, glycerol distearate, without or with wax, or other substances well known in the art. this field of technology.

The compounds of formula (I), and/or at least one pharmaceutically acceptable salt thereof, may also be administered, for example, intravenously, subcutaneously and/or intramuscularly in any pharmaceutically acceptable and suitable injectable form. Typical injectable forms include, but are not limited to, sterile aqueous solutions containing acceptable carriers and solvents such as, for example, water, Ringer's solution, and isotonic sodium chloride solution; sterile oil-in-water microemulsions and aqueous or oil suspensions.

Preparations for parenteral administration may be in the form of aqueous or non-aqueous isotonic sterile injectable solutions or suspensions. These solutions and suspensions can be prepared from sterile powders or granules using one or more carriers or diluents used in oral preparations, or using other suitable dispersing or wetting agents and suspending agents. The compounds of the present invention can be dissolved in water, polyethylene glycol, propylene glycol, ethanol, corn oil, cottonseed oil, peanut oil, sesame oil, benzyl alcohol, sodium chloride, gum tragacanth and/or various buffering agents. Other adjuvants and administration methods are well known in the pharmaceutical industry. The active ingredient may also be administered by injection in the form of a composition with suitable carriers including saline, dextrose or water, or with cyclodextrin (namely Captisol), substances suitable for solubilization in a co-solvent system (namely propylene glycol ), or substances suitable for micellar solubilization (namely with Tween 80).

The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example a solution in 1,3-butanediol. Water, Ringer's solution and isotonic sodium chloride solution may be used as acceptable carriers and solvents. In addition, sterile non-volatile oils are usually used as a solvent or suspending medium. Any neutral fixed oil can be used for this purpose, including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used to obtain injectable preparations.

A sterile injectable oil-in-water microemulsion can be prepared, for example, by 1) dissolving at least one compound of formula (I) in an oil phase, such as, for example, a mixture of soybean oil and lecithin; 2) mixing the oil phase containing the compound of formula (I) with a mixture of water and glycerin and 3) stirring this mixture to obtain a microemulsion.

A sterile aqueous or oily suspension can be prepared in accordance with techniques known in the art. For example, a sterile aqueous solution or suspension can be prepared using a non-toxic parenterally acceptable diluent or solvent such as, for example, 1,3-butanediol; and the sterile oily suspension can be prepared using a sterile non-toxic acceptable solvent or sterile non-toxic acceptable suspending medium, such as, for example, sterile fixed oils, such as synthetic mono- or diglycerides, and fatty acids, such as, for example, oleic acid.

Pharmaceutically acceptable carriers, adjuvants and excipients that can be used in the pharmaceutical compositions of the present invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, self-emulsifying drug delivery systems (SEDDS) such as d-alpha tocopherol polyethylene glycol 1000 succinate, surfactants used in pharmaceutical dosage forms such as Tweens, polyethoxylated castor oil such as CREMOPHOR surfactant (BASF), or other similar polymeric delivery matrices, serum proteins such as human serum albumin , buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, mixtures of partial esters of glycerol with saturated fatty acids of plant origin, water, salts or electrolytes such as protamine sulfate, sodium hydrogen orthophosphate, potassium hydrogen orthophosphate, sodium chloride, zinc salts, colloid silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene block polymers, polyethylene glycol and lanolin. In addition, cyclodextrins such as alpha, beta and gamma cyclodextrin, or chemically modified derivatives thereof such as hydroxyalkyl cyclodextrins, including 2- and 3 -hydroxypropyl-cyclodextrins, or other solubilized derivatives.

The pharmaceutically active compounds of the present invention can be processed in accordance with standard pharmaceutical techniques to produce medicinal agents for administration to patients, including humans and other mammals. These pharmaceutical compositions may be subjected to standard processing steps used in the pharmaceutical field, such as sterilization, and/or may contain standard adjuvants such as preservatives, stabilizers, wetting agents, emulsifiers, buffering agents, and so on. Additionally, enteric coatings can be applied to tablets and pills. Such compositions may also contain adjuvants, for example wetting agents, sweetening agents, flavoring agents and flavoring agents.

The amounts of compounds administered and the mode of administration when treating a disease state using the compounds and/or compositions of the present invention depend on a number of factors, including the age, weight, sex, health status of the subject, type of disease, severity of disease, route and frequency of administration, and the particular compound used . Accordingly, the mode of administration may vary widely, but can be routinely determined using standard techniques. A suitable daily dose may be from about 0.001 to 100 mg/kg body weight, preferably from about 0.0025 to about 50 mg/kg body weight, and most preferably from about 0.005 to 10 mg/kg body weight. The daily dose can be administered in the form of 1-4 fractional doses per day. Other administration schedules include one dose per week and one dose every two days.

For therapeutic purposes, the active compounds of the present invention are typically used in combination with one or more adjuvant suitable for the specified route of administration. Compounds intended for oral administration may be mixed with lactose, sucrose, starch powder, cellulose alkanoic acid esters, cellulose alkyl esters, talc, stearic acid, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphorus and sulfuric acid, gelatin, gum acacia, sodium alginate, polyvinylpyrrolidone and/or polyvinyl alcohol and then tabletted or encapsulated for ease of administration. Such capsules or tablets may contain a controlled release formulation in the form of a dispersion of the active compound in hydroxypropyl methylcellulose.

The pharmaceutical compositions of the present invention contain at least one compound of formula (I), and/or at least one pharmaceutically acceptable salt of said compound, and optionally an additional agent selected from any pharmaceutically acceptable carrier, adjuvant and excipient. Alternative compositions of the present invention contain a compound of formula (I) described herein, or a prodrug thereof, and a pharmaceutically acceptable carrier, adjuvant or excipient.

Application.

The compounds of formula (I) can be used for the treatment of cancer.

According to another embodiment, the present invention provides the preparation of a combination of a compound of formula (I), and/or a pharmaceutically acceptable salt of the said compound, a stereoisomer of the said compound, or a tautomer of the said compound, and additional therapeutic agent(s) to simultaneously , their separate or sequential administration for the treatment and/or prevention of numerous diseases or disorders associated with inhibition of the DGK target in T cells.

According to another aspect of the present invention, there is provided a method of treating a patient suffering from, or predisposed to, a pathological condition that is associated with inhibition of a DGK target in T Cells. Some of the pathological conditions can be cured. The proposed method involves administering to a patient a therapeutically effective amount of a composition containing a compound of formula (I), and/or a pharmaceutically acceptable salt of said compound, a stereoisomer of said compound, or a tautomer of said compound. For example, the compounds described herein can be used to treat or prevent viral infections and proliferative diseases such as cancer.

The compounds of formula (I) and pharmaceutical compositions containing at least one compound of formula (I) can be used for the treatment or prevention of any diseases or pathological conditions that are associated with inhibition of the DGK target in T cells. These diseases and conditions include viral and other infections (eg, skin infections, GI infections, urinary tract infections, genitourinary infections, systemic infections) and proliferative diseases (eg, cancer). The compounds of formula (I) and pharmaceutical compositions containing at least one compound of formula (I) can be administered to animals, preferably mammals (eg, domesticated animals, cats, dogs, mice, rats) and more preferably humans. Any administration technique may be used to deliver a given compound or pharmaceutical composition to a patient. According to some embodiments of the present invention, a compound of formula (I) or a pharmaceutical composition containing at least a compound of formula (I) is administered orally. According to other embodiments of the present invention, a compound of formula (I) or a pharmaceutical composition containing at least a compound of formula (I) is administered parenterally.

Compounds of formula (I) can inhibit the activity of diacylglycerol kinases alpha and zeta (DGKa/Z. For example, compounds of formula (I) can be used to inhibit the activity of DGKa and DGKZ, in cells or in a subject requiring modulation of DGKa and DGKZ, by administering an inhibitory amount a compound of formula (I) or a salt of said compound.

Additionally, the present invention provides methods for treating diseases associated with the activity or expression, including abnormal activity and/or increased expression, of DGKa and DGKZ, in a subject (eg, a patient) by administering to the subject in need of such treatment a therapeutically effective amount or dose a compound of formula (I) or a pharmaceutical composition containing said compound. Examples of such diseases may include any disease, disorder or pathological condition that is directly or indirectly associated with the expression or activity of the DGKa and DGKZ enzymes, such as overexpression or abnormal activity. DGKa- and DGKZ-associated disease can also include any disease, disorder or pathological condition that can be prevented, ameliorated or treated by modulating the activity of the DGKa and DGKZ enzymes. Examples of DGKa- and DGKZ-associated diseases include cancer and viral infections such as HIV infection, hepatitis B and hepatitis C.

According to one aspect of the present invention, sequential administration involves that

- 16 044500 the compound(s) of formula (I) is administered before the administration of the immuno-oncology agent. According to another aspect of the present invention, the compound(s) of Formula (I) is administered simultaneously with an immuno-oncology agent. According to another aspect of the present invention, sequential administration involves the compound(s) of formula (I) being administered after administration of the immuno-oncology agent.

According to another aspect of the present invention, the compounds of formula (I) may be presented in the form of a preparation further containing an immuno-oncology agent.

Immuno-oncology agents include, for example, a small molecule drug, antibody, or other biological or small molecule. Examples of biological immuno-oncology agents include, but are not limited to, cancer vaccines, antibodies, and cytokines. According to one aspect of the present invention, the antibody is a monoclonal antibody. According to another aspect of the present invention, the monoclonal antibody is a humanized antibody or a human antibody.

According to one aspect of the invention, the immuno-oncology agent is (i) a stimulatory (including co-stimulatory) receptor agonist or (ii) an antagonist of inhibitory (including co-inhibitory) T cell signaling, both of which (often called immune checkpoint regulators) cause enhancement of antigen-specific T-cell responses.

Some stimulatory and inhibitory molecules belong to the immunoglobulin superfamily (IgSF). One important family of membrane-bound ligands that bind to costimulatory or coinhibitory receptors is the B7 family, including B7-1, B7-2, B7-H1 (PD-L1), B7-DC (PD-L2), B7-H2 ( ICOS-L), B7-H3, B7-H4, B7-H5 (VISTA) and B7-H6. Another family of membrane-bound ligands that bind to co-stimulatory or co-inhibitory receptors is the TNF family of molecules that bind to cognate receptors belonging to the TNF receptor family, these ligands and receptors include CD40 and CD40L, OX-40, OX-40L, CD70, CD27L, CD30, CD30L, 4-1BBL, CD137 (4-1BB), TRAIL/Apo2-L, TRAILR1/DR4, TRAILR2/DR5, TRAILR3, TRAILR4, OPG, RANK, RANKL, TWEAKR/Fn14, TWEAK, BAFFR, EDAR , XEDAR, TACI, APRIL, BCMA, LTeR LIGHT, DcR3, HVEM, VEGI/TL1A, TRAMP/DR3, EDAR, EDA1, XEDAR, EDA2, TNFR1, lymphotoxin α/TNFe, TNFR2, TNFa, LTeR, lymphotoxin α1β2, FAS, FASL, RELT, DR6, TROY, NGFR.

According to one aspect of the present invention, T cell responses can be stimulated by a combination of a compound of formula (I) and one or more agents selected from (i) a protein antagonist that inhibits T cell activation (e.g., an immune checkpoint inhibitor) , such as CTLA-4, PD-1, PD-L1, PD-L2, AG-3, TIM-3, galectin 9, SEACAM-1, BTLA, CD69, galectin-1, TIGIT, CD113, GPR56, VISTA, 2B4, CD48, GARP, PD1H, LAIR1, TIM-1 and TIM-4, and (ii) a protein agonist that stimulates T cell activation such as B7-1, B7-2, CD28, 4-1BB (CD137) , 41BBL, ICOS, ICOS-L, OX40, OX40L, GITR, GITRL, CD70, CD27, CD40, DR3 and CD28H.

Other agents that can be used in combination with compounds of formula (I) for the treatment of cancer include antagonists of inhibitory receptors located on NK cells (natural killer cells), or agonists of activating receptors located on NK cells. For example, compounds of formula (I) can be used in combination with KIR (killer cell immunoglobulin-like receptor) antagonists such as lirilumab.

Other combination therapy agents include agents that inhibit or deplete macrophages or monocytes, including but not limited to CSF-1R (colony stimulating factor receptor) antagonists, such as anti-CSF-1R antagonist antibodies, including RG7155 (WO 11/70024, WO 11/107553, WO 11/131407, WO 13/87699, WO 13/119716, WO 13/132044) or FPA-008 (WO 11/140249; WO 13169264; WO 14/036357).

According to another aspect of the present invention, the compounds of formula (I) can be used in conjunction with one or more agonist agent that binds positive co-stimulatory receptors, a blocking agent that attenuates signaling through inhibitory receptors, an antagonist, and one or more agents which systemically increases the frequency of anti-cancer T cells, an agent that bypasses various immunosuppressive pathways in the tumor microenvironment (eg, blocks inhibitory receptor engagement (eg, PD-L1/PD-1 interaction), depletes or inhibits Tregs (regulatory T cells) (e.g., an anti-CD25 monoclonal antibody (e.g., daclizumab) or depletion using bead-linked antibodies against ex vivo CD25-positive cells may be used here), inhibits metabolic enzymes such as IDO (indoleamine 2,3-dioxygenase ), or eliminates/prevents anergy and T cell exhaustion), and an agent that triggers activation of the innate immune response and/or inflammation at the tumor site.

According to one aspect of the present invention, the immuno-oncology agent is an antagonist of the CTLA-4 protein, such as an anti-CTLA-4 antagonist antibody. Suitable anti-CTLA-4 antibodies include, for example, YERVOY (ipilimumab) or tremelimumab.

According to another aspect of the present invention, the immuno-oncology agent is a PD-1 (programmed cell death protein 1) antagonist, such as an anti-PD-1 antagonist antibody. Suitable anti-PD-1 antibodies include, for example, OPDIVO (nivolumab), KEYTRUDA (pembrolizumab) or MEDI-0680 (AMP-514; WO 2012/145493). These immuno-oncology agents may also include pidilizumab (CT-011), although the specificity of its binding to PD-1 is questionable. Another approach to target the PD-1 receptor is to use the recombinant protein AMP-224, consisting of the extracellular domain of the PD-L2 protein (B7-DC) fused to the Fc fragment of IgG1.

According to another aspect of the present invention, the immuno-oncology agent is an antagonist of the PD-L1 protein, such as an anti-PD-L1 antagonist antibody. Suitable anti-PD-L1 antibodies include, for example, MPDL3280A (RG7446; WO2010/077634), durvalumab (MEDI4736), BMS-936559 (WO 2007/005874) and MSB0010718C (WO 2013/79174).

According to another aspect of the present invention, the immuno-oncology agent is a LAG-3 (lymphocyte activation gene 3) protein antagonist, such as an anti-LAG-3 antagonist antibody. Suitable anti-LAG3 antibodies include, for example, BMS-986016 (WO 10/19570, WO 14/08218) or IMP-731 or IMP-321 (WO 08/132601, WO 09/44273).

According to another aspect of the present invention, the immuno-oncology agent is a CD137 receptor agonist (4-1BB), such as an anti-CD137 agonist antibody. Suitable anti-CD137 antibodies include, for example, urelumab and PF-05082566 (WO 12/32433).

According to another aspect of the present invention, the immuno-oncology agent is a GITR (glucocorticoid-inducible receptor superfamily of tumor necrosis factor receptor) agonist, such as an anti-GITR agonist antibody. Suitable anti-GITR antibodies include, for example, BMS-986153, BMS-986156, TRX-518 (WO 06/105021, WO 09/009116) and MK-4166 (WO 11/028683).

According to another aspect of the present invention, the immuno-oncology agent is an IDO antagonist. Suitable IDO antagonists include, for example, INCB-024360 (WO 2006/122150, WO 07/75598, WO 08/36653, WO 08/36642), indoximod, BMS-986205 or NLG-919 (WO 09/73620, WO 09/ 1156652, WO 11/56652, WO 12/142237).

According to another aspect of the present invention, the immuno-oncology agent is an OX40 receptor agonist, such as an anti-OX40 agonist antibody. Suitable anti-OX40 antibodies include, for example, MEDI-6383 or MEDI-6469.

According to another aspect of the present invention, the immuno-oncology agent is an OX40L ligand antagonist, such as an anti-OX40 antagonist antibody. Suitable OX40L ligand antagonists include, for example, RG-7888 (WO 06/029879).

According to another aspect of the present invention, the immuno-oncology agent is a CD40 receptor agonist, such as an anti-CD40 agonist antibody. According to another embodiment of the present invention, the immuno-oncology agent is a CD40 receptor antagonist, such as an anti-CD40 antagonist antibody. Suitable anti-CD40 antibodies include, for example, lucatumumab or dacetuzumab.

According to another aspect of the present invention, the immuno-oncology agent is a CD27 receptor agonist, such as an anti-CD27 agonist antibody. Suitable anti-CD27 antibodies include, for example, varlilumab.

According to another aspect of the present invention, the immuno-oncology agent is MGA271 (to B7H3) (WO 11/109400).

Combination therapy is intended to involve sequential administration of these therapeutic agents, i.e. administering the therapeutic agents at different times, as well as substantially simultaneous administration of the therapeutic agents, or at least two of the therapeutic agents. Substantially simultaneous administration may be accomplished, for example, by administering to the subject a single single-administration dosage form containing all therapeutic agents in a fixed proportion, or multiple single-administration dosage forms each containing one therapeutic agent. Sequential or substantially simultaneous administration of therapeutic agents can be accomplished by any suitable route, including, without limitation, oral routes of administration, intravenous routes of administration, intramuscular routes of administration, and direct absorption through mucosal tissues. The therapeutic agents may be administered by the same route or by different routes. For example, the first therapeutic agent of a selected combination may be administered by intravenous injection, while other therapeutic agents of the combination may be administered orally. Alternatively, for example, all therapeutic agents may be administered orally, or all therapeutic agents may be administered by intravenous injection. Combination therapy may also include administration of the therapeutic agents described above in another combination with other biologically active ingredients and non-drug treatment (eg, surgery or radiation therapy). When the combination therapy additionally includes a non-drug treatment, the non-drug treatment can be administered at any appropriate time until a positive result is achieved from the combination of therapeutic agents and the non-drug treatment. For example, in appropriate cases, a positive result may persist for several days or even weeks when non-drug treatment is temporarily excluded and only the administration of therapeutic agents remains.

As used herein, the term cell is intended to refer to a cell that is in vitro, ex vivo, or in vivo. In some embodiments of the present invention, the ex vivo cell may be part of a tissue sample removed from an organism, such as a mammal. In some embodiments of the present invention, the in vitro cell may be a cell in cell culture. In some embodiments of the present invention, the in vivo cell is a cell living in an organism, such as a mammal.

As used herein, the term contacting refers to the association of said moieties in an in vitro system or an in vivo system. For example, contacting the enzyme DGKa and DGKZ with a compound of formula (I) involves administering a compound of the present invention to an individual or patient, for example a person having DGKa and DGKZ, and also, for example, administering a compound of formula (I) to a sample containing cellular or purified a preparation containing the enzyme DGKa and DGKZ.

The term DGKa and DGKZ enzyme inhibitor refers to an agent capable of inhibiting the activity of diacylglycerol kinase alpha and/or diacylglycerol kinase zeta (DGKa and DGKZ) in T cells, resulting in stimulation of the T cells. The DGKa and DGKZ enzyme inhibitor may be a reversible or irreversible inhibitor of the DGKa and DGKZ enzymes. A reversible inhibitor of DGKa and DGKZ enzymes is a compound that reversibly inhibits the activity of DGKa and DGKZ enzymes at a catalytic site or a non-catalytic site, and an irreversible inhibitor of DGKa and DGKZ enzymes is a compound that irreversibly disrupts the activity of DGKa and DGKZ enzymes by forming a covalent bond between inhibitor and enzyme.

Types of cancer that can be treated using a compound of formula (I) include, but are not limited to, brain cancers, skin cancers, bladder cancers, ovarian cancers, breast cancers, gastric cancers, pancreatic cancers , prostate malignancies, colon malignancies, blood malignancies, lung malignancies and bone malignancies. Examples of such types of cancer include neuroblastoma, intestinal carcinoma such as rectal carcinoma, colon carcinoma, familial adenomatous polyposis carcinoma and hereditary nonpolyposis colorectal cancer, esophageal carcinoma, labial carcinoma, laryngeal carcinoma, subpharyngeal carcinoma, tongue carcinoma, salivary carcinoma glands, gastric carcinoma, adenocarcinoma, medullary thyroid carcinoma, papillary thyroid carcinoma, renal carcinoma, renal parenchymal carcinoma, ovarian carcinoma, cervical carcinoma, uterine body carcinoma, endometrial carcinoma, chorionic carcinoma, pancreatic carcinoma, prostate carcinoma, testicular carcinoma , breast carcinoma, urinary organ carcinoma, melanoma, brain tumors such as glioblastoma, astrocytoma, meningioma, medulloblastoma, and peripheral neuroectodermal tumors, Hodgkin's lymphoma, non-Hodgkin's lymphoma, Burkitt's lymphoma, acute lymphatic leukemia (ALL), chronic lymphatic leukemia ( CLL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), adult T-cell leukemia/lymphoma, diffuse large B-cell lymphoma (DLBCL), hepatocellular carcinoma, gallbladder carcinoma, bronchial carcinoma, small cell lung carcinoma, non-small cell lung carcinoma, multiple myeloma, basal cell carcinoma, teratoma, retinoblastoma, uveal melanoma, seminoma, rhabdomyosarcoma, craniopharyngioma, osteosarcoma, chondrosarcoma, myosarcoma, liposarcoma, fibrosarcoma, Ewing's sarcoma and plasmacytoma.

One or more additional pharmaceutical agents or therapeutic modalities, such as, for example, antiviral agents, chemotherapeutic or other anticancer agents, immunostimulants, immunosuppressants, ionizing radiation, antitumor and antiviral vaccines, cytokine therapy (eg, IL2 and GM-CSF), and /or tyrosine kinase inhibitors, may optionally be used in combination with compounds of formula (I) for the treatment of DGKa- and DGKZ-associated diseases, disorders or pathological conditions. These agents may be used in combination with the compounds of the present invention in a single dosage form, or the agents may be administered simultaneously or sequentially in separate dosage forms.

Suitable chemotherapeutic or other anticancer agents include, for example, alkylating agents (including, without limitation, nitrogen mustards, ethyleneimine derivatives, alkylsulfonates, nitrosoureas and triazenes) such as uramustine, chlormethine, cyclophosphamide (CY-19 044500

ΤΟΧΑΝζ), ifosfamide, melphalan, chlorambucil, pipobromane, triethylene-melamine, triethylenethiophosphoramine, busulfan, carmustine, lomustine, streptozocin, dacarbazine and temozolomide.

Agents suitable for use in combination with compounds of formula (I) for the treatment of melanoma include: dacarbazine (DTIC), optionally together with other chemotherapeutic drugs such as carmustine (BCNU) and cisplatin; the Dartmouth regimen, which consists of DTIC, BCNU, cisplatin, and tamoxifen; a combination of cisplatin, vinblastine and DTIC, temozolomide or YERVOY™. The compounds of formula (I) in the treatment of melanoma can also be used in combination with immunotherapy drugs, including cytokines such as interferon alpha, interleukin 2 and tumor necrosis factor (TNF).

The compounds of formula (I) can also be used in combination with vaccine therapy for the treatment of melanoma. Melanoma vaccines are somewhat similar to viral vaccines, which are used to prevent diseases caused by viruses such as polio, measles and mumps. Weakened melanoma cells or parts of melanoma cells, called antigens, may be injected into the patient to stimulate the body's immune system to destroy the melanoma cells.

For the treatment of melanomas located on the arms and legs, a combination of agents comprising one or more compounds of formula (I) in combination with an isolated hyperthermic limb perfusion technique can also be used. This treatment protocol involves temporarily separating the circulation of the manipulated limb from that of the rest of the body and injecting high doses of chemotherapy drugs into the artery supplying that limb, thereby delivering high doses to the tumor site without exposing internal organs. given doses that might otherwise cause severe side effects. Usually the liquid is heated to 38.9-40.0°C. Melphalan is the drug most commonly used in this chemotherapy technique. Another agent called tumor necrosis factor (TNF) may also be used.

Suitable chemotherapeutic or other anticancer agents include, for example, antimetabolites (including, without limitation, folate antagonists, pyrimidine analogs, purine analogs, and adenosine deaminase inhibitors) such as methotrexate, 5-fluorouracil, floxuridine, cytarabine, 6-mercaptopurine, 6-thioguanine , fludarabine phosphate, pentostatin and gemcitabine.

Suitable chemotherapeutic or other anticancer agents further include, for example, certain natural products and their derivatives (eg, vinca alkaloids, antitumor antibiotics, enzymes, lymphokines, and epipodophyllotoxins), such as vinblastine, vincristine, vindesine, bleomycin, dactinomycin, daunorubicin, doxorubicin, epirubicin , idarubicin, araC, paclitaxel (Taxol), mithramycin, deoxycoformycin, mitomycin-C, L-asparaginase, interferons (especially IFN-a), etoposide and teniposide.

Other cytotoxic agents include navelbene, CPT-11, anastrazole, letrazole, capecitabine, reloxafine and droloxafine.

Suitable cytotoxic agents also include cytotoxic agents such as epidophyllotoxin; antitumor enzyme; topoisomerase inhibitor; procarbazine; mitoxantrone; platinum coordination complexes such as cisplatin and carboplatin; biological response modifiers; growth inhibitors; antihormonal therapeutic agents; leucovorin; tegafur and hematopoietic growth factors.

Other anticancer agents include therapeutic antibodies such as trastuzumab (HERCEPTIN®), antibodies against costimulatory molecules such as CTLA-4, 4-1BB and PD-1, or antibodies against cytokines (IL-1O or TGF-β).

Other anticancer agents also include anticancer agents that block the migration of immune cells, such as chemokine receptor antagonists, including CCR2 (C-C chemokine receptor type 2) and CCR4 (C-C chemokine receptor type 4).

Other anticancer agents also include anticancer agents that enhance the immune system, such as adjuvants or T cell adoptive transfer immunotherapy.

Cancer vaccines include dendritic cells, synthetic peptides, DNA vaccines, and recombinant viruses.

The pharmaceutical composition according to the invention may optionally include at least one signal transduction inhibitor (STI). A signal transduction inhibitor is an agent that selectively inhibits one or more steps of signaling pathways critical for the normal functioning of cancer cells, thereby inducing their apoptosis. Suitable STIs include, but are not limited to: (i) bcr/abl kinase inhibitors, such as, for example, STI 571 (GLEEVECZ); (ii) epidermal growth factor (EGF) receptor inhibitors, such as, for example, kinase inhibitors (IRESAZ, SSI-774) and antibodies (Imclone: C225 [Goldstein et al., Clin. Cancer Res., 1:13111318 (1995) ] and Abgenix: ABX-EGF); (iii) her-2/neu receptor inhibitors, such as farnesyltransferase inhibitors (FTI), such as, for example, L-744832 (Kohl et al., Nat. Med., 1(8):792-797 (1995)) ; (iv) inhibitors of the Akt family of kinases or the Akt pathway, such as, for example, rapamycin (see, for example, Sekulic et al., Cancer Res., 60:3504-3513 (2000)) ; (v) cell cycle kinase inhibitors, such as, for example, flavopiridol and UCN-O1 (see, for example, Sausville, Curr. Med. Chem. Anti-Canc. Agents, 3:47-56 (2003)); and (vi) phosphatidylinositol kinase inhibitors, such as, for example, LY294002 (see, for example, Vlahos et al, J. Biol. Chem., 269:5241-5248 (1994)). Alternatively, at least one STI and at least one compound of formula (I) may be presented in separate pharmaceutical compositions. According to a specific embodiment of the present invention, at least one compound of formula (I) and at least one STI can be administered to a patient simultaneously or sequentially. In other words, at least one compound of formula (I) may be administered first, at least one STI may be administered first, or at least one compound of formula (I) and at least one STI may be administered simultaneously. In addition, when more than one compound of formula (I) and/or STI is used, the compounds may be introduced in any order.

The present invention further provides a pharmaceutical composition for the treatment of a chronic viral infection in a patient, comprising at least one compound of formula (I), optionally at least one chemotherapeutic drug and optionally at least one antiviral agent in a pharmaceutically acceptable carrier.

A method of treating a chronic viral infection in a patient by administering an effective amount of the above-mentioned pharmaceutical composition is also provided.

According to a specific embodiment of the present invention, at least one compound of formula (I) and at least one chemotherapeutic agent are administered to a patient simultaneously or sequentially. In other words, at least one compound of formula (I) may be administered first, at least one chemotherapeutic agent may be administered first, or at least one compound of formula (I) and at least one STI may be administered simultaneously. In addition, when more than one compound of formula (I) and/or a chemotherapeutic agent is used, the compounds may be administered in any order. Likewise, any antiviral agent or STI can also be administered at any time relative to the administration of the compound of formula (I).

Chronic viral infections that can be treated with the combination therapy of the present invention include, but are not limited to, those caused by hepatitis C virus (HCV), human papillomavirus (HPV), cytomegalovirus (CMV), herpes simplex virus (HSV), Epstein-Barr (EBV), varicella zoster virus, Coxsackie virus, human immunodeficiency virus (HIV). Notably, the above-mentioned methods can also be used to treat parasitic infections (eg, malaria), which optionally use already known compounds suitable for treating parasitic infections instead of antiviral agents.

Suitable antiviral agents considered as candidates for use in combination with a compound of formula (I) may include nucleoside and nucleotide reverse transcriptase inhibitors (NRTIs), non-nucleoside reverse transcriptase inhibitors (NNRTIs), protease inhibitors and other antiviral drugs.

Examples of suitable NRTIs include zidovudine (AZT); didanosine (ddl); zalcitabine (ddC); stavudine (d4T); lamivudine (3TC); abacavir (1592U89); adefovir dipivoxil [bis(ROM)-RMEA]; lobucavir (BMS-180194); BCH-I0652; emitricitabine [(-)-FTC]; beta-L-FD4 (also called beta-b-O4C and called beta-L-2',3'-dicleoxy-5-fluoro-cytidene); DAPD, ((-)-beta-O-2,6-diaminopurine dioxolane) and lodenosine (FddA). Typical suitable NNRTIs include nevirapine (BI-RG-587); delavirdine (VNAR, U-90152); efavirenz (DMP-266); PNU-142721; AG-1549; MKC-442 (1-(ethoxymethyl)-5-(1methylethyl)-6-(phenylmethyl)-(2,4(1H,3H)-pyrimidinedione) and (+)-calanolide A (NSC-675451) and B. Typical Suitable protease inhibitors include saquinavir (Ro 31-8959); ritonavir (ABT-538); indinavir (MK-639); nelfnavir (AG-1343); amprenavir (141W94); lasinavir (BMS-234475); DMP- 450; BMS-2322623; ABT-378 and AG-1549 Other antiviral agents include hydroxyurea, ribavirin, IL-2, IL-12, pentafuside and Yissum Project No. 11607.

The present invention also includes pharmaceutical kits that can be used, for example, for the treatment or prevention of DGKa- and DGKZ-associated diseases or disorders and other diseases mentioned herein, including one or more containers containing a pharmaceutical composition containing a therapeutically effective amount of compound of formula (I). Such kits may further include, if desired, one or more different standard pharmaceutical components used in the kits, such as, for example, containers with one or more pharmaceutically acceptable carriers, additional containers, as will be readily apparent to those skilled in the art. . The kit may also include instructions, in the form of inserts or labels, indicating quantities of components to be administered, recommendations for administration, and/or recommendations for mixing components.

- 21 044500

Combination therapy is intended to involve sequential administration of these therapeutic agents, i.e. administering the therapeutic agents at different times, as well as substantially simultaneous administration of the therapeutic agents, or at least two of the therapeutic agents. Substantially simultaneous administration may be accomplished, for example, by administering to the subject a single single-administration dosage form containing all therapeutic agents in a fixed proportion, or multiple single-administration dosage forms each containing one therapeutic agent. Sequential or substantially simultaneous administration of therapeutic agents can be accomplished by any suitable route, including, without limitation, oral routes of administration, intravenous routes of administration, intramuscular routes of administration, and direct absorption through mucosal tissues. The therapeutic agents may be administered by the same route or by different routes. For example, the first therapeutic agent of a selected combination may be administered by intravenous injection, while other therapeutic agents of the combination may be administered orally. Alternatively, for example, all therapeutic agents may be administered orally, or all therapeutic agents may be administered by intravenous injection. Combination therapy may also include administration of the therapeutic agents described above in another combination with other biologically active ingredients and non-drug treatment (eg, surgery or radiation therapy). When the combination therapy further includes a non-drug treatment, the non-drug treatment can be administered at any appropriate time until a positive result is achieved from the combination of therapeutic agents and the non-drug treatment. For example, in appropriate cases, a positive result may persist for several days or even weeks when non-drug treatment is temporarily excluded and only the administration of therapeutic agents remains.

The present invention also provides pharmaceutically acceptable compositions that contain a therapeutically effective amount of one or more compounds of formula (I) together with one or more pharmaceutically acceptable carrier and/or diluent and optionally one or more an additional therapeutic agent described above.

The compounds of the invention, in any of their uses described herein, may be administered in any suitable manner, for example orally, i.e. in the form of tablets, capsules (each of which includes sustained-release or extended-release preparations), pills, powders, granules, elixirs, tinctures, suspensions (including nanosuspensions, microsuspensions, spray-drying dispersions), syrups and emulsions; sublingual; buccal; parenterally, for example, using subcutaneous, intravenous, intramuscular or intrasternal injection or infusion techniques (for example, in the form of sterile injectable aqueous or non-aqueous solutions or suspensions); nasally, including administration into the nasal mucosa, for example using an inhalation spray; topically, for example in the form of a cream or ointment; or rectally, for example in the form of suppositories. The compounds of the invention may be administered by themselves, but are typically administered in conjunction with a pharmaceutical carrier selected taking into account the route of administration used and standard pharmaceutical practice.

The phrase pharmaceutically acceptable carrier as used herein means a pharmaceutically acceptable substance, composition or excipient, such as a liquid or solid filler, diluent, excipient, adjuvant, used in the manufacturing process of a drug (for example, lubricant, talc, magnesium, calcium or zinc stearate or stearic acid), or a substance encapsulating the solvent involved in the transfer or transport of the compound according to the invention from one organ or part of the body to another organ or part of the body. Each carrier must be acceptable in terms of compatibility with the other ingredients of the formulation, including an adjuvant, excipient or excipient, such as diluents, preservatives, excipients, flow control agents, disintegrating agents, wetting agents, emulsifying agents, suspending agents, sweetening agents, flavoring agents, flavoring agents, antibacterial agents, antifungal agents, lubricating agents and distributing agents, depending on the nature of the route of administration and dosage forms, and harmless to the patient.

The term pharmaceutical composition means a composition containing a compound of the invention in combination with at least one additional pharmaceutically acceptable carrier.

Pharmaceutically acceptable carriers are included in the formulation, taking into account a number of factors within the competence of those of ordinary skill in the art. These factors include, without limitation, the type and nature of the active agent included in the formulation; the characteristics of the subject to whom the agent-containing composition will be administered; the intended route of administration of this composition and target therapeutic indications. Pharmaceutically acceptable carriers include aqueous and non-aqueous liquid vehicles, as well as a variety of solid and semi-solid dosage forms. Such media can

- 22 044500 include several different ingredients and excipients in addition to the active agent, such additional ingredients included in the preparation for a variety of reasons, for example to stabilize the active agent, binders and so on, are well known to those of ordinary skill in the art. Descriptions of suitable pharmaceutically acceptable carriers and factors influencing their selection can be found in many readily available sources, such as, for example, Allen, L.V. et al., Remington: The Science and Practice of Pharmacy (2 Volumes), 22nd Edition (2012), Pharmaceutical Press.

The mode of administration of the compounds of the present invention will, of course, vary depending on such known factors as the pharmacodynamic characteristics of the particular agent and the mode and route of administration; biological species to which the recipient belongs, its age, gender, health status, medical condition and body weight; the nature and severity of symptoms; type of concurrent treatment; frequency of treatment repetition; route of administration, functional parameters of the patient's kidneys and liver, and the desired effect.

As a general guideline, the daily oral dosage of each active ingredient required to achieve a therapeutic effect will generally range from about 0.001 to about 5000 mg per day, preferably in the range from about 0.01 to about 1000 mg per day, and most preferably ranging from about 0.1 to about 250 mg per day. When administered intravenously, the most preferred doses are from about 0.01 to about 10 mg/kg/min at a constant infusion rate. The compounds of the present invention may be administered once daily as a single dose, or the total daily dose may be administered in divided doses of two, three or four divided doses.

The compounds are typically administered in mixtures with suitable pharmaceutical diluents, excipients or carriers (collectively referred to herein as pharmaceutical carriers) selected based on the intended form of administration, which may be, for example, oral tablets, capsules, elixirs, syrups and the like. , and in accordance with standard pharmaceutical practice.

Suitable dosage forms (pharmaceutical compositions) for administration may contain from about 1 milligram to about 2000 milligrams of active ingredient per unit dosage form. In these pharmaceutical compositions, the active ingredient is typically present in an amount of approximately 0.1-95% (by weight) of the total weight of the composition.

A typical oral capsule contains at least one of the compounds of the present invention (250 mg), lactose (75 mg) and magnesium stearate (15 mg). This mixture is passed through a 60 mesh sieve and filled into a gelatin capsule No. L.

A typical injectable preparation is prepared by aseptically placing at least one of the compounds of the present invention (250 mg) in a vial, then lyophilizing and aseptically sealing. Before use, the contents of this tube are mixed with 2 ml of saline to obtain an injectable drug.

Included within the scope of the present invention are pharmaceutical compositions containing as an active ingredient at least one of the compounds of the present invention, in a therapeutically effective amount, either alone or in combination with a pharmaceutical carrier. Optionally, the compounds of the present invention may be used alone, in combination with other compounds of the invention, or in combination with one or more other therapeutic agents, eg, an anticancer agent, or other pharmaceutically active agent.

Regardless of the route of administration chosen, the compounds of the present invention, which can be administered in a suitable hydrated form, and/or the pharmaceutical compositions of the present invention are prepared in pharmaceutically acceptable dosage forms using standard techniques known to those skilled in the art.

The actual dosage levels of the active ingredients in the pharmaceutical compositions of the present invention may vary in order to obtain an amount of the active ingredient that will be effective in achieving the desired therapeutic response in a particular patient and depending on the composition and route of administration, but in any case not must be toxic to the patient.

The dose level selected will generally depend on a number of factors, including the activity of the particular compound of the present invention used, or the ester, salt or amide of the compound used, route of administration, time of administration, rate of excretion or metabolism of the particular compound used, rate and extent of absorption, duration of treatment, other drugs, compounds and/or substances used in combination with the particular compound used, age, sex, body weight, pathological condition, general health and medical history of the patient being treated, and other similar factors well known in the medical field.

A physician or veterinarian of ordinary skill in the art can readily determine and prescribe an effective amount of the required pharmaceutical composition. For example, a physician or veterinarian might begin by prescribing a lower dosage level of the compounds of the invention used in a pharmaceutical composition than that required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved.

In general, a suitable daily dose of a compound according to the present invention is that amount of the compound that is the lowest dose that produces a therapeutic effect. The level of such effective dose will generally depend on the factors described above. Typically, doses of the compounds of the present invention for oral, intravenous, intracerebroventricular and subcutaneous administration to a patient range from about 0.01 to about 50 mg per kilogram of body weight per day.

If desired, an effective daily dose of the active compound may be administered in two, three, four, five, six or more sub-doses, which are administered separately throughout the day at suitable intervals, optionally in unit dosage forms. In some aspects of the present invention, the entire daily dose is administered at one time.

Although the compound of the present invention can be administered by itself, it is still preferable when the compound is administered in the form of a pharmaceutical preparation (composition).

The above other therapeutic agents, when used in combination with the compounds of the present invention, can be used, for example, in the amounts specified in the Physician's Desk Reference (PDR), otherwise these amounts can be determined by one of ordinary skill in the art. In the methods of the present invention, such other therapeutic agent (or agents) may be administered before, simultaneously with, or after the compounds of the invention.

Synthesis techniques.

The compounds of the present invention can be synthesized using many techniques available to those skilled in the art of organic chemistry. General synthetic routes used to prepare the compounds of the present invention are described below. These diagrams are illustrative and are not intended to limit the use of other possible techniques that one skilled in the art may employ to prepare the compounds described herein. Those skilled in the art will appreciate that a variety of techniques can be used to prepare the compounds of the present invention. Examples of compounds of the present invention prepared using the procedures described in the general schemes are given in the examples section below. The preparation of homochiral compounds can be accomplished using techniques known to one skilled in the art. For example, homochiral compounds can be prepared by resolving racemic products or diastereoisomers using chiral preparative HPLC. Alternatively, the compounds described in the Examples section can be synthesized using known techniques that provide enantiomerically or diastereomerically enriched products.

The reactions and procedures described in this section are performed in solvents suitable for the reagents and substances used, and these reactions and procedures are also suitable for achieving the desired transformations. In addition, in the description of the synthesis procedures given below, it is understood that all proposed reaction conditions, including the choice of solvent, reaction atmosphere, temperature of the reaction mixture, duration of the experiment and procedures for isolating the reaction product, are selected in accordance with standard conditions for this reaction, which without labor must be understood by a specialist in this field of technology. One skilled in the art of organic synthesis will appreciate that the functional groups present in various parts of the molecule must be compatible with the proposed reagents and reactions. The limitations placed on substituents that are compatible with the reaction conditions will be readily apparent to one skilled in the art, and alternative approaches are required when incompatible substituents are present. Therefore, in order to obtain the desired compound of the present invention, it is sometimes necessary to change the order of synthetic steps or to favor one particular synthetic route over another. It should also be appreciated that another important factor when planning any synthetic route in the art is the judicious choice of the protecting group used to protect the reactive functional groups present in the compounds described in this invention. For qualified practitioners, an authoritative source containing a wide range of alternatives to these groups is Wuts and Greene, Greene's Protective Groups in Organic Synthesis, Fourth Edition, Wiley and Sons (2007).

Examples

The following examples are provided to illustrate specific and preferred embodiments of the present invention and do not limit the scope of the present invention. Chemical abbreviations and symbols and scientific abbreviations and symbols have their usual and customary meanings unless otherwise indicated. Additional abbreviations used in the examples section and elsewhere in this invention are defined above. Common Intermediates

- 24 044500 can usually be used for synthesis in more than one example, these compounds are identified in the order they appear (for example, intermediate 1, intermediate 2, etc.), and the abbreviations Int are used to designate them. 1 or II, Int. 2 or I2 and so on. Example compounds are identified using the example number and the name of the step where they were prepared (e.g., 1-A means Example 1, step A), or using the Example number alone, the compound being the compound listed in the example title (e.g. 1 means the connection indicated in the title of example 1). In some cases, alternative procedures for preparing intermediates or example compounds are described. Often those skilled in the art of chemical synthesis may develop alternative preparation procedures that may be desirable when one or more factors must be taken into account, such as shorter reaction time, lower consumption of starting materials, ease of operation or isolation, higher yield, more productive catalysis, elimination of toxic reagents, availability of specialized equipment and reduction in the number of linear stages, and so on. The description of alternative synthetic procedures is intended to provide additional options for the preparation of exemplary compounds of the present invention. In some cases, certain functional groups in the presented examples and claims may be replaced using known bioisosteric substitutions known in the art, for example, by replacing the carboxyl group with tetrazole or phosphate group. ^{The 1} H-NMR spectrum was recorded in deuterated dimethyl sulfoxide; water signals were suppressed during data processing. In the spectra shown, the effects of suppression of water signals are not corrected. Proton signal intensities near the frequency used to suppress the water signal (3.35 ppm) are reduced.

- 25 044500

Abbreviations Ac acetyl without anhydrous water WATER aza-HOBt 7-aza-1-hydroxybenzotriazole Bn benzyl 1 -BOC-piperazine m/eth-butyl piperazine-1-carboxylate Bu butyl CV column volume DCE dichloroethane DXM dichloromethane DEA diethylamine DIEA diisopropylethylamine (Hunig's base) DIPEA diisopropylethylamine DMA UA-dimethylacetamide DMF dimethylformamide DMSO dimethyl sulfoxide E.A. ethyl acetate EDC 1 -(3-dimethylaminopropyl)-3 ethylcarbodiimide hydrochloride Et ethyl hour, hours or h watch) HATU (1 -[bis(dimethylamino)methylene]-\H-1,2,3triazolo[4,5-b]pyridinium 3-oxide, hexafluorf osph at) HCl hydrochloric acid HPLC high performance liquid chromatography KHMDS potassium bis(trimethylsilyl)amide LC liquid chromatography LC-MS liquid chromatography-mass spectrometry M molar mm millimolar Me methyl MHz megahertz min minute(s)

- 26 044500

M ⁺¹ (M+N) ⁺ ms mass spectrometry n. or N normal NaHMDS sodium bis(trimethylsilyl)amide N.B.S. Y-bromosuccinimide nM nanomolar NMP U-methylpyrrolidinone Ph phenyl PYBROP Bromotripyrrolidinophosphonium hexafluorophosphate RuPhos, precatalyst chloro(2-dicyclohexylphosphino-2',6'diisopropoxy-1,1'-biphenyl) [2-(2'-amino-1, G biphenyl)]palladium (II) RT or Ret-time retention time us. saturated tre from-VnON TEA tertiary butanol triethylamine TFU trifluoroacetic acid THF tetrahydrofuran those thin layer chromatography ROS phosphorus oxychloride Xphos 2nd generation CAS No. 1310584-14-5

Intermediate 1: 2-(methylthio)-3-nitro-1,5-naphthyridin-4-ol

Ethyl 3-aminopicolinate (1.71 g, 10.29 mmol) and (2-nitroethene-1,1-diyl)bis(methylsulfane) (1.75 g, 10.59 mmol) were mixed without solvent and melted at 130 °C. After 44 h, the reaction mixture was cooled to room temperature and stirred with excess diethyl ether. The resulting orange solid was collected on filter paper (1.7 g, 68%). ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.75 (d, J=4.6 Hz, 1H), 8.53 (d, J=8.6 Hz, 1H), 7.99 (dd, J=8.4, 5.0 Hz, 1H) , 2.59 (s, 3H).

Intermediate 2: 4-chloro-2-(methylthio)-3-nitro-1,5-naphthyridine

2-(Methylthio)-3-nitro-1,5-naphthyridin-4-ol (1.5 g, 6.32 mmol) and phosphorus oxychloride (8.84 mL, 95 mmol) were mixed at room temperature and then heated with stirring at 50°C overnight. The reaction mixture was then cooled to room temperature and water was slowly added. The resulting orange solid was collected on filter paper and dried under vacuum to yield intermediate 2 (1 g, 62%). 1H NMR (400 MHz, chloroform-d) δ 9.06 (d, J=3.9 Hz, 1H), 8.35 (d, J=8.3 Hz, 1H), 7.79 (dd, J=8.6, 4.2 Hz, 1H), 2.77 (s, 3H). Analytical LC-MS conditions: column: Phenomenex Luna C18, 2.0x50 mm, particle size 3.0 µm; mobile phase A: (5:95) methanol:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) methanol:water with 0.1% trifluoroacetic acid; gradient: 0-100% B for 4 min, then maintaining 100% B for 0.75 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: 3.7 min, 256, 258 (M+H).

Intermediate 3: 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-2-(methylthio)-3nitro-1,5-naphthyridine

- 27 044500

1-(Bis(4-fluorophenyl) methyl)piperazine (0.992 g, 3.44 mmol) and triethylamine (0.436 ml, 3.13 mmol). This reaction mixture was stirred for 1 hour at room temperature. Completion of the reaction was determined by LC-MS analysis. The solvent was removed by rotary evaporation. The residue was purified by silica gel chromatography using ethyl acetate and hexanes to give the product (0.7 g, 44%) as a yellow solid. 1H NMR (400 MHz, chloroform-d) δ 8.71 (dd, J= 4.2, 1.7 Hz, 1H), 8.15 (dd, J=8.6, 1.7 Hz, 1H), 7.56 (dd, J=8.6, 4.2 Hz, 1H), 7.47-7.37 (m, 4H), 7.07-6.96 (m, 4H), 5.32 (s, 1H), 4.34 (s, 1H), 3.73-3.60 (m, 4H), 2.69-2.63 (m, 7H). LC-MS: Analytical LC-MS conditions: Column: Phenomenex Luna C18, 2.0x50 mm, particle size 3.0 µm; mobile phase A: (5:95) methanol:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) methanol:water with 0.1% trifluoroacetic acid; gradient: 0-100% B for 4 min, then maintaining 100% B for 0.75 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: 4.1 min, 508 (M+H).

Intermediate 4: 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-3-nitro-1,5-naphthyridin-2(III)-one

4-(4-(bis(4-Fluorophenyl)methyl)piperazin-1-yl)-2-(methylthio)-3-nitro-1,5-naphthyridine (0.69 g, 1.359 mmol) was dissolved in acetic acid ( 13.59 ml) to obtain a yellow suspension. Hydrogen peroxide (0.166 mL, 1.631 mmol) and sodium tungstate dihydrate (0.067 g, 0.204 mmol) were added successively and the mixture was stirred overnight at room temperature. Acetic acid was removed by rotary evaporation and the residue was chromatographed on silica gel with 0-80% ethyl acetate in hexanes. A white solid was isolated (400 mg, 62%). 1H NMR (400 MHz, chloroform-d) δ 8.54-8.48 (m, 1H), 7.70-7.60 (m, 1H), 7.50-7.45 (m, 1H), 7.41 (dd, J=8.8, 5.4 Hz, 4H ), 7.02 (t, J= 8.7 Hz, 4H), 4.34 (s, 1H), 3.72 (br. s., 4H), 2.65 (d, J=4.4 Hz, 4H). Analytical LC-MS conditions: Column: Phenomenex Luna C18, 2.0x50 mm, particle size 3.0 µm; mobile phase A: (5:95) methanol:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) methanol:water with 0.1% trifluoroacetic acid; gradient: 0-100% B for 4 min, then maintaining 100% B for 0.75 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: 3.3 min, 478 (M+H).

Intermediate 5: 4-(4-benzhydrylpiperazin-1-yl)-2-(methylthio)-3-nitro-1,5-naphthyridine

4-Chloro-2-(methylthio)-3-nitro-1,5-naphthyridine (1 g, 3.91 mmol) and 1-benzhydrylpiperazine (0.987 g, 3.91 mmol) were dissolved in dichloromethane (19.56 ml) in a round bottom flask. Triethylamine (0.545 mL, 3.91 mmol) was added and the reaction mixture was stirred for 1 hour at room temperature. LC-MS analysis indicated that the reaction was complete. The solvent was removed by rotary evaporation. The residue was purified by silica gel chromatography using ethyl acetate and hexanes to give the title compound (1.5 g, 3.2 mmol, yield: 81%) as a yellow solid. ¹ H NMR (400 MHz, chloroform-d) δ 8.66 (dd, J=4.1, 1.6 Hz, 1H), 8.10 (dd, J=8.4, 1.6 Hz, 1H), 7.52 (dd, J=8.5, 4.0 Hz , 1H), 7.48-7.40 (m, 4H), 7.30-7.25 (m, 4H), 7.20-7.15 (m, 2H), 4.30 (s, 1H), 3.69-3.60 (m, 4H), 2.67-2.61 (m, 4H), 2.59 (s, 3H). Analytical LC-MS conditions: Column: Phenomenex Luna C18,

- 28 044500

2.0x50 mm, particle size 3.0 microns; mobile phase A: (5:95) methanol:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) methanol:water with 0.1% trifluoroacetic acid; gradient: 0-100% B for 4 min, then maintaining 100% B for 0.75 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: 3.6 min, 472 (M+H).

Intermediate 6: 4-(4-Benzhydrylpiperazin-1-yl)-3-nitro-1,5-naphthyridin-2(1H)-one

In a round bottom flask, 4-(4-benzhydrylpiperazin-1-yl)-2-(methylthio)-3-nitro-1,5-naphthyridine (1.5 g, 3.18 mmol) was dissolved in acetic acid (31.8 ml ) to obtain a yellow suspension. Hydrogen peroxide (0.389 mL, 3.82 mmol) was added followed by sodium tungstate dihydrate (0.157 g, 0.477 mmol). The resulting suspension was stirred overnight at room temperature and then heated for 72 hours at 30°C. LC-MS analysis showed a single peak corresponding to the desired product. Volatiles were removed under high vacuum to give the title compound (1.4 g, 3.2 mmol, yield: 78%). 1H NMR (400 MHz, DMSO-d ₆ ) δ 8.51 (d, J=4.2 Hz, 1H), 7.71-7.66 (m, 1H), 7.60 (dd, J=7.9, 4.3 Hz, 1H), 7.48 (d , J=7.6 Hz, 4H), 7.32 (t, J=7.3 Hz, 4H), 7.25-7.17 (m, 2H), 4.42 (s, 1H), 3.51 (d, J=4.2 Hz, 4H), 2.58 -2.52 (m, 4H hidden by residual DMSO). Analytical LC-MS conditions: column: Waters BEH C18, 2.0x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 40°C; gradient: maintaining 0% B for 0.5 min, 0-100% B for 4 min, then maintaining 100% B for 0.5 min; flow rate: 1 ml/min. Analytical LC-MS results confirmed the title compound: 3.5 min, 440 (M-H), 442 (M+H).

Intermediate 7: 6-bromo-4-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile

CH ₃

6-Bromo-1-methyl-2,4-dioxo-1,2,3,4-tetrahydro-1,5naphthyridine-3-carbonitrile (2.50 g, 8.93 mmol) was added to a 500 ml round bottom flask. acetonitrile (89 ml) and DIEA (9.4 ml, 53.8 mmol) was added. This reaction mixture was stirred for two minutes. After adding DIEA, the reaction mixture looked like a homogeneous solution. Phosphorus oxychloride (3.3 mL, 35.4 mmol) was added to the reaction mixture, and then benzyltriethylammonium chloride (2.68 g, 11.77 mmol) was added. This reaction mixture was stirred under nitrogen atmosphere overnight at room temperature to obtain a dark brownish heterogeneous mixture. Volatile components were removed under vacuum using a system consisting of a rotary evaporator and a vacuum pump. Ice and 1.5 M potassium hydrogen orthophosphate solution were added to the remainder of the reaction mixture, and the mixture was partitioned between dichloromethane and chloroform. The aqueous fraction was extracted with chloroform. The combined organic extracts were washed with potassium hydrogen orthophosphate, 1N. HCl and then a mixture of potassium hydrogen orthophosphate and saline solution. The organic fractions were dried over sodium sulfate, filtered and the solvent was removed in vacuo using a rotary evaporator to give a brown solid (~3.1 g). The resulting solid was redissolved in chloroform/dichloromethane and adsorbed onto silica gel (9.8 g). The substance was then purified by chromatography (suspension of silica gel (83 g) in 2% ethyl acetate in dichloromethane), using 2% ethyl acetate in dichloromethane for elution. Fractions containing product were combined and the solvent was removed in vacuo to give 1.922 g (72%) of the title compound. 1H NMR (chloroform-d) δ 7.81 (d, J=8.8 Hz, 1H), 7.66 (d, J=9.0 Hz, 1H), 3.75 (s, 3H). Analytical LC-MS conditions: Waters Acquity UPLC VEN C18, 2.1x 50 mm, particle size 1.7 µm; mobile phase A: 100% water with 0.05% trifluoroacetic acid; mobile phase B: 100% acetonitrile with 0.05% trifluoroacetic acid; temperature: 40°C; gradient: 298% B for 1.5 min, then maintain 98% B for 0.5 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. Analytical LC-MS results confirmed the title compound: 1.3 min, 298, 300 (M+H).

Intermediate 8: tert-butyl 4-([1,1'-biphenyl]-2-ylmethyl)piperazine-1-carboxylate

- 29 044500

2-Phenylbenzyl bromide (2.264 g, 9.16 mmol) was dissolved in DMF (70 ml). 1-BOCpiperazine (1.717 g, 9.22 mmol) was then added and then potassium carbonate (1.272 g, 9.20 mmol) was added. The reaction vessel was closed. The reaction mixture was stirred overnight (21 hours) at room temperature. Volatile components were removed from the reaction mixture under vacuum using a rotary evaporator. The reaction mixture was then partitioned between ethyl acetate and water. The aqueous phase was extracted with ethyl acetate (1x). The organic extracts were combined, washed with saline and dried over magnesium sulfate. The drying agent was filtered off and the solvent was removed from the resulting filtrate under vacuum using a rotary evaporator to give Intermediate 8 as a clear oil. LC-MS: column: Waters Acquity VEN 2.1 mm x 50 mm, particle size 1.7 µm; mobile phase A: (10:90) acetonitrile:water with 0.1% TFA; mobile phase B: (90:10) acetonitrile:water with 0.1% TFA; temperature: 40°C; gradient: from 0% B to 100% B for 2 min, then maintaining 100% B for 1 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Injected sample volume: 1 µl. Retention time = 1.24 min; observed adducts: [M+H]; measured masses: 353.3. ¹ H NMR (αcetonitrile-d ₃ ) δ 7.51 (dd, J=7.1, 1.6 Hz, 1H), 7.28-7.45 (m, 7H), 7.21-7.28 (m, 1H), 3.37 (s, 2H), 3.24 -3.32 (m, 4H), 2.18-2.26 (m, 4H), 1.39 (s, 9H).

Intermediate 9: 1-([1,1'-biphenyl]-2-ylmethyl)piperazine, TFA

tert-Butyl 4-([1,1'-biphenyl]-2-ylmethyl)piperazine-1-carboxylate (3.219 g, 9.13 mmol) was dissolved in dichloromethane (25 ml), then TFA (50.0 ml) was added . This reaction mixture was placed under a nitrogen atmosphere and stirred for 2 hours at room temperature. Volatile components were removed from the reaction mixture under vacuum using a rotary evaporator. Toluene was added to the resulting reaction product and volatile components were removed under vacuum to facilitate removal of excess TFA. Dissolution in toluene and removal of volatile components was repeated again. Ethyl acetate, hexanes and diethyl ether were added to convert the product from an oil to a solid to give a semi-solid, colorless precipitate. After removing the solvents and drying the product under vacuum at room temperature, Intermediate 9 was obtained as a colorless solid. LC-MS: column: Waters Acquity VEN 2.1 mm x 50 mm, particle size 1.7 µm; mobile phase A: (10:90) acetonitrile:water with 0.1% TFA; mobile phase B: (90:10) acetonitrile:water with 0.1% TFA; temperature: 40°C; gradient: from 0% B to 100% B for 2 min, then maintaining 100% B for 1 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Injected sample volume: 1 µl. Retention time = 0.92 min; observed adducts: [M+H]; measured masses: 253.2. 1H NMR (DMSO-d ₆ ) δ 8.65 (br. s., 2H), 7.52-7.62 (m, 1H), 7.34-7.51 (m, 7H), 7.20-7.32 (m, 1H), 3.67 (br. s., 2H), 3.08 (br. s., 4H), 2.52-2.71 (m, 4H).

Intermediate 10: tert-butyl 4-(naphthalene-1-ylmethyl)piperazine-1-carboxylate

Intermediate 1-1OA: 1-(iodomethyl)naphthalene

- 30 044500

1-(Chloromethyl)naphthalene (1.5 g, 8.49 mmol) was dissolved in acetone (54 ml). Sodium iodide (1.52 g, 10.14 mmol) was added to this reaction mixture. The reaction mixture was heated to reflux for 1.25 hours. The reaction mixture was then cooled and filtered through a pad of diatomaceous earth (Celite). The solvent was removed from the resulting filtrate using a rotary evaporator. The resulting reaction product was dissolved in diethyl ether and filtered through another pad of diatomaceous earth (Celite) to remove remaining salts. The solvent was again removed under vacuum using a rotary evaporator to obtain 2.25 g of 1-(iodomethyl)naphthalene as an amber solid. Purity assessed by NMR gave a value of 80-85%. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ 8.16-8.13 (m, 1.0H), 7.99 (d, J=8.2 Hz, 1.0H), 7.91 (d, J=8.4 Hz, 1.0H), 7.74 -7.71 (m, 1.2H), 7.707.66 (m, 1.2H), 7.56 (ddd, J=8.1, 6.9, 1.1 Hz, 1.1H), 7.42 (dd, J=8.1,7.2 Hz, 1.0H) , 5.13 (s, 2.0H).

Intermediate connection 10.

1-(Iodomethyl)naphthalene (2.248 g, 8.39 mmol) was dissolved in DMF (70 ml) and 1-BOC-piperazine (1.586 g, 8.52 mmol). Potassium carbonate (1.164 g, 8.42 mmol) was added. The reaction vessel was closed. The reaction mixture was stirred for 2 hours at room temperature. The solvent was removed under vacuum using a rotary evaporator and the remainder of the reaction mixture was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate. The organic extract was washed successively with a mixture of water and saline solution and then with saline solution only. The organic extract was then dried over magnesium sulfate. The drying agent was filtered off and the solvent was removed in vacuo to give an orange oil. The resulting orange oil was purified by silica gel column chromatography, eluting with a gradient of 0-10% ethyl acetate in dichloromethane. The purified product fractions were combined to obtain 2.46 g of tert-butyl 4-(naphthalene-1ylmethyl)piperazine-1-carboxylate as a colorless oil. LC-MS: Column: Waters Acquity VEN

C18 2.1 mmx50 mm, particle size 1.7 microns; mobile phase A: 100% water with 0.05% TFA; mobile phase B: 100% acetonitrile with 0.05% TFA; temperature: 40°C; gradient: from 2% B to 98% B for 1.5 min, then maintaining 98% B for 0.5 min; flow rate: 0.8 ml/min; registration: MS and UV (220 nm). Injected sample volume: 3 µl. Retention time = 0.88 min; observed adducts: [M+H]; measured masses: 327.2. 1H NMR (acetonitrile-d ₃ ) δ 8.28-8.35 (m, 1H), 7.87-7.92 (m, 1H), 7.80-7.85 (m, 1H), 7.48-7.57 (m, 2H), 7.41-7.47 (m , 2H), 3.90 (s, 2H), 3.25-3.40 (m, 4H), 2.34-2.48 (m, 4H), 1.42 (s, 9H).

Intermediate 11: 1-(naphthalene-1-ylmethyl)piperazine at ^ΠΝΗ (Ι-P) tert-Butyl 4-(naphthalene-1-ylmethyl)piperazine-1-carboxylate (2.41 g, 7.38 mmol) dissolved in dichloromethane (37 ml). TFA (37.0 ml) was then added. The reaction vessel was sealed under a nitrogen atmosphere. The reaction mixture was stirred for 1.5 hours at room temperature. Toluene (70 ml) was added to the reaction mixture and volatile components were removed under vacuum. Dichloromethane and toluene were added to this mixture. Volatiles were removed under vacuum to facilitate removal of excess TFA. The desired product was obtained as a creamy beige solid. LC-MS: column:

Waters Acquity VEN C18 2.1 mmx50 mm, particle size 1.7 microns; mobile phase A: 100% water with 0.05% TFA; mobile phase B: 100% acetonitrile with 0.05% TFA; temperature: 40°C; gradient: from 2% B to 98% B for 1.5 min, then maintaining 98% B for 0.5 min; flow rate: 0.8 ml/min; registration: MS and UV (220 im). Injected sample volume: 3 µl. Retention time = 0.68 min; observed adducts: [M+H]; measured masses: 227.1. 1H NMR (DMSO^) δ 8.73 (br. s., 2H), 8.28 (d, J=8.2 Hz, 1H), 7.87-8.06 (m, 2H), 7.39-7.69 (m, 4H), 4.26 (br . s., 2H), 3.17 (br. s., 4H), 2.91 (br. s., 3H).

Intermediate 12. Ethyl 3-amino-6-bromopicolinate <^/ ^NH2

L,L/O_sn ₃

Br Ν γ ^J

O (1-12)

Ethyl 3-aminopicolinate (8.0 g, 48.1 mmol) was suspended in water (66 mL) in a 250 mL three-neck round bottom flask equipped with a mechanical stirrer, addition funnel, and temperature probe with thermocouple. Sulfuric acid (1.7 mL, 31.9 mmol) and acetic acid (3.31 mL, 57.8 mmol) were slowly added while immersing the flask in a room temperature water bath to maintain temperature. A solution of bromine (2.5 mL, 48.5 mmol) in acetic acid (17.5 mL, 306 mmol) was then added to the reaction mixture over 15 min at ambient temperature with vigorous stirring, maintaining the internal temperature of the reaction mixture below 23 °C. Oud water bath

- 31 044500 was poured and the reaction mixture was stirred for 2 hours at ambient temperature. This reaction suspension was filtered and washed with a small amount of water and then dried under vacuum at room temperature to obtain 9.305 g of Intermediate 12 as a yellow solid. LC-MS: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: 100% water with 0.05% trifluoroacetic acid; mobile phase B: 100% acetonitrile with 0.05% trifluoroacetic acid; temperature: 40°C; gradient: 2-98% B for 1.5 min, then maintain 98% B for 0.5 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. Retention time = 0.94 min; observed adducts: [M+H]; measured masses: 245.0. 1H NMR (DMSO-d ₆ ) δ 7.44 (d, J=8.8 Hz, 1H), 7.21 (d, J=8.7 Hz, 1H), 6.88 (br. s., 2H), 4.29 (q, J=7.1 Hz, 2H), 1.31 (t, J=7.1 Hz, 3H).

Intermediate 13: Ethyl 3-acetamido-6-bromopicolinate

Ethyl 3-amino-6-bromopicolinate (1.31 g, 5.35 mmol) was dissolved in THF (6 ml) and then acetic anhydride (1.6 ml, 16.96 mmol) was added. This reaction mixture was a suspension/incomplete solution. The reaction mixture was placed under a nitrogen atmosphere and heated to reflux. The reaction mixture became homogeneous within 15 minutes. The reaction mixture was refluxed for 4 hours. Volatile components of the reaction mixture were removed under vacuum using a rotary evaporator. A small amount of ethyl acetate was added to the remainder of the reaction mixture and a nearly colorless solid was collected by filtration, which was dried under vacuum to give 787 mg of Intermediate 13. LC-MS: Column: Waters Acquity UPLC VEN C18, 2.1 x 50 mm, particle size 1, 7 µm; mobile phase A: 100% water with 0.05% trifluoroacetic acid; mobile phase B: 100% acetonitrile with 0.05% trifluoroacetic acid; temperature: 40°C; gradient: 2-98% B for 1.5 min, then maintain 98% B for 0.5 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. Retention time = 0.98 min; observed adducts: [M+H]; measured masses: 287.0. 1H NMR (DMSO-d ₆ ) δ 10.40 (s, 1H), 8.32 (d, J=8.7 Hz, 1H), 7.83 (d, J=8.8 Hz, 1H), 4.33 (q, J=7.1 Hz, 2H ), 2.12 (s, 3H), 1.32 (t, J=7.2 Hz, 3H). Removal of solvent from the filtrate provided an additional 695 mg of product (purity: 87%).

Intermediate 14: Ethyl 3-acetamido-6-cyanopicolinate

Copper(I) cyanide (86 mg, 0.960 mmol) and ethyl 3-acetamido-6-bromopicolinate (200 mg, 0.697 mmol) were added to a 2.5 mL microwavable tube (Biotage) and then N,Ndimethylformamide (3 .5 ml). A magnetic stir bar was added and the tube was sealed under a nitrogen atmosphere. This reaction mixture was heated to 200°C for 8 minutes in a Biotage Initiator microwave synthesizer in normal adsorption mode. The reaction mixture was then transferred to a round bottom flask and the solvent was removed under vacuum using a system consisting of a vacuum pump and a rotary evaporator. The resulting brown residue was triturated with ethyl acetate (25 ml) containing DCM and the resulting suspension was filtered through a pad of diatomaceous earth (Celite). The solvent was removed from the filtrate to give the crude product as a yellow solid (102 mg). This crude product was purified on a Biotage Isolera One system using an Isco RediSep silica gel cartridge (4 g) and eluting with a gradient of 1-10% ethyl acetate in dichloromethane. Fractions containing pure product were combined and the solvent was removed in vacuo to obtain 84 mg of pure product as a colorless solid. LC-MS: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: 100% water with 0.05% trifluoroacetic acid; mobile phase B: 100% acetonitrile with 0.05% trifluoroacetic acid; temperature: 40°C; gradient: 2-98% B for 1.5 min, then maintain 98% B for 0.5 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. Retention time = 0.92 min; observed adducts: [M+H]; measured masses: 234.1. 1H NMR (DMSO-dg) δ 10.67 (s, 1H), 8.58 (d, J=8.7 Hz, 1H), 8.19 (d, J=8.7 Hz, 1H), 4.36 (q, J=7.1 Hz, 2H) , 2.17 (s, 3H), 1.33 (t, J=7.1 Hz, 3H).

Intermediate 15: Ethyl 6-cyano-3-(N-methylacetamido)picolinate

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Ethyl 3-acetamido-6-cyanopicolinate (20 mg, 0.086 mmol) was dissolved in DMF (0.85 ml). Cesium carbonate (39.1 mg, 0.120 mmol) was then added and then methyl iodide (9 μL, 0.144 mmol) was added. The reaction vessel was closed and the reaction mixture was stirred overnight at room temperature.

The reaction mixture was then diluted with acetonitrile (2.8 mL) and filtered through a 0.45 μm syringe filter. Volatiles were removed from the resulting filtrate, then the filtrate was triturated with DCM/ethyl acetate and filtered again through a 0.45 μm syringe filter to remove salts. Volatiles were removed under vacuum using a rotary evaporator to obtain the crude product as an amber oil. The resulting oil was dissolved in deuterated chloroform to record the proton NMR spectrum. The proton NMR spectrum contained characteristic features indicating limited rotation (rotamer): 1H NMR (chloroform-d) δ 7.65-8.01 (m, 2H), 4.32-4.56 (m, 2H), 3.43 (br. s., 1H ), 3.22 (s, 2H), 2.26 (br. s., 1H), 1.82 (s, 2H), 1.34-1.50 (m, 3H). The spectra obtained in ^{the 13} C DEPT experiment (undistorted enhancement by polarization transfer experiment) corresponded to N-methylation with a signal of 37 ppm.

The NMR sample was reduced by removing volatile components under vacuum using a rotary evaporator and then the sample was dissolved in 1 ml of DMF/acetonitrile (1:1). This crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; gradient: 0-30% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. The product yield was 17.9 mg and its purity, determined by LC-MS analysis, was 99%. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Results: retention time = 1.11 min; observed adducts: [M+H]; measured masses: 248.0. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Retention time =1.01 min; observed adducts: [M+H]; measured masses: 248.0.

Intermediate 16: 8-hydroxy-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile

CH ₃

HE (1-16)

Potassium bis(trimethylsilyl)amide (KHMDS) (3.0 mL, 1,500 mmol) in 0.5 M toluene was added to a 25 mL round bottom flask. The flask was placed under a nitrogen atmosphere and cooled to -78°C. A solution of ethyl 6-cyano-3-(Nmethylacetamido)picolinate (333 mg, 1.347 mmol) in THF (13.5 ml) was added via cannula to the KHMDS solution over approximately 23 min. This reaction mixture was kept at -78°C for 20 minutes, then the dry ice bath was removed and the reaction mixture was warmed to room temperature over 1.25 hours. Ethyl acetate and water were added to the reaction mixture. The reaction mixture was then transferred to a separatory funnel and additional ethyl acetate was added. The reaction mixture was shaken gently and then separated, collecting the aqueous phase in a small Erlenmeyer flask. The aqueous phase (~20 ml) was acidified using 1.6 ml of 1N. hydrochloric acid solution. The fine yellow precipitate was collected by filtration, washed with deionized water (~2 ml) and dried under vacuum at room temperature to obtain 229 mg of intermediate 16 as a yellow solid. LC-MS: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: 100% water with 0.05% trifluoroacetic acid; mobile phase B: 100% acetonitrile with 0.05% trifluoroacetic acid; temperature: 40°C; gradient: 2-98% B for 1.5 min, then maintain 98% B for 0.5 min; flow rate: 0.8 ml/min; registration:

- 33 044500

UV, 220 nm. Retention time = 0.68 min; observed adducts: [M+H]; measured masses: 202.1. 1H

NMR (DMSO-d ₆ ) δ 11.51 (br. s., 1H), 8.21 (d, J=8.8 Hz, 1H), 8.11 (d, J=8.8 Hz, 1H), 6.16 (s, 1H), 3.54 (s, 3H).

Intermediate 17: 8-hydroxy-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2carbonitrile

CH ₃

HE (1-17)

Acetic acid (2.8 ml) was added to a 2 dram test tube containing 8-hydroxy-5-methyl-6-oxo-5,6-dihydro-1,5naphthyridine-2-carbonitrile (227 mg, 1.128 mmol), stir and add nitric acid (0.151 mL, 3.39 mmol) slowly. This mixture was left to stir for approximately one minute, then the tube was capped and the mixture was heated to 100°C for 18 minutes. The reaction mixture became homogeneous after approximately 3 minutes at 100°C. The mixture was cooled, then placed in an ice bath and then the yellow precipitate that formed was collected by filtration and washed with cold ethanol (2x0.5 ml). This product was dried under vacuum at room temperature to obtain the title compound (202 mg) as a yellow solid. LC-MS: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: 100% water with 0.05% trifluoroacetic acid; mobile phase B: 100% acetonitrile with 0.05% trifluoroacetic acid; temperature: 40°C; gradient: 2-98% B for 1.5 min, then maintain 98% B for 0.5 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. Retention time = 0.67 min; observed adducts: [M+H]; measured masses: 247.1. 1H NMR (DMSO) δ 8.30-8.38 (m, 1H), 8.19-8.28 (m, 1H), 3.59 (s, 3H).

Intermediate 18: 8-chloro-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2carbonitrile

CH ₃

NC^ ⁴ NO ₂

Cl (1-18)

A magnetic stir bar and acetonitrile were added to a 2 dram test tube containing 8-hydroxy-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5naphthyridine-2-carbonitrile (192 mg, 0.780 mmol) (3.1 ml). DIEA (0.272 mL, 1.560 mmol) was then added to the resulting suspension. This reaction mixture was stirred for 1-2 minutes until the reaction mixture took the form of a homogeneous yellow solution. Phosphorus oxychloride (0.131 mL, 1.404 mmol) was added to the reaction mixture. The test tube was capped in a nitrogen atmosphere with gas supplied through an oil bubbler. The reaction mixture was stirred for 1.5 hours at room temperature, then benzyltriethylammonium chloride (200 mg, 0.878 mmol) was added to the reaction mixture. The tube was capped under a nitrogen atmosphere, immersed in an oil bath (65°C) and heated for 1 hour. The reaction mixture was then cooled and volatile components were removed from the reaction mixture under vacuum using a rotary evaporator. The remainder of the reaction mixture was dissolved in ethyl acetate, poured into a beaker containing ice (~10 ml) and then transferred to a separatory funnel. The aqueous phase was extracted with ethyl acetate. The organic extracts were combined and washed successively with 1.5 M K2HPO4 solution, saturated aqueous sodium bicarbonate solution and saline solution. The organic extract was dried over magnesium sulfate, filtered and the solvent was removed in vacuo to obtain 204 mg of a brownish crystalline solid. LC-MS: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: 100% water with 0.05% trifluoroacetic acid; mobile phase B: 100% acetonitrile with 0.05% trifluoroacetic acid; temperature: 40°C; gradient: 2-98% B for 1.5 min, then maintain 98% B for 0.5 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. Retention time = 1.01 min; observed adducts: [M+H]; measured masses: 265.0 (weak ionization). ^1H NMR (chloroform-d) δ 8.03 (d, J=8.8 Hz, 1H), 7.89-7.97 (m, 1H), 3.82 (s, 3H).

Intermediate 19: 6-bromo-4-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3carbonitrile

CH ₃

J t t Br^N^y^CN

Cl (1-19)

6-Bromo-1-methyl-2,4-dioxo-1,2,3,4-tetrahydro-1,5-naphthyridine-3-carbonitrile (2.50 g, 8.93 mmol) was suspended in acetonitrile (89 ml ). DIEA (9.4 mL, 53.8 mmol) was then added and the mixture was stirred for two minutes. Phosphorus oxychloride (POCl ₃ ) (3.3 ml,

- 34 044500

35.4 mmol) and then benzyltriethylammonium chloride (2.68 g, 11.77 mmol) was added. The reaction mixture was placed under a nitrogen atmosphere and stirred for 18 hours at room temperature. The volatile components were then removed from the reaction mixture under vacuum using a system consisting of a rotary evaporator and a vacuum pump. Ice was added to the remainder of the reaction mixture, then a 1.5 M solution of potassium hydrogen orthophosphate was added. Dichloromethane and chloroform were then added. This mixture was transferred to a separatory funnel to mix and separate the aqueous and organic phases. The aqueous phase was extracted with chloroform. The organic extracts were combined and washed successively with a 1.5 M solution of potassium hydrogen orthophosphate, 1 N. a solution of hydrochloric acid and a mixture of potassium hydrogen orthophosphate and saline solution. The organic extract was dried over sodium sulfate, then filtered and the solvents were removed from the filtrate under vacuum to obtain a brown solid. This product was purified using silica gel column chromatography, eluting with 2% ethyl acetate in dichloromethane. Fractions containing pure product by TLC analysis were combined and the solvent was removed in vacuo to give the title compound (1.684 g) as a yellow solid. LC-MS: column: Waters Acquity VEN 2.1 mm x 50 mm, particle size 1.7 µm; mobile phase A: (10:90) acetonitrile:water with 0.1% TFA; mobile phase B: (90:10) acetonitrile:water with 0.1% TFA; temperature: 40°C; gradient: from 0% B to 100% B for 2 min, then maintaining 100% B for 1 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Injected sample volume: 1 µl. Retention time = 1.29 min; observed adducts: [M+H]; measured masses: 298.1. Ή NMR (chloroform-d) δ 7.81 (d, J=8.8 Hz, 1H), 7.66 (d, J=9.0 Hz, 1H), 3.75 (s, 3H).

Intermediate 20: tert-butyl 4-(6-bromo-3-cyano-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridin-4-yl)piperazin-1-carboxylate

CH ₃ dXX

VG Ν γ CN o N ₃ C ^Χ Ν ⁿ 3S (1-20)

6-Bromo-4-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (545 mg, 1.826 mmol) was dissolved in DMF (18 ml). 1-BOC-piperazine (354 mg, 1.901 mmol) and potassium carbonate (381 mg, 2.76 mmol) were then added. This reaction mixture was stirred at room temperature under a nitrogen atmosphere. After approximately 1 hour, the reaction mixture became heterogeneous and became a suspension. This suspension was stirred for 5.5 hours. According to HPLC analysis, the reaction was 99% complete. DMF (9 ml) was added to the reaction mixture and the mixture was stirred overnight. Ethyl acetate and 1.0 M potassium hydrogen orthophosphate (pH ~ 4.5) were then added. THF was added. This mixture was placed in a separating funnel and some of the aqueous phase was drained, the remaining mixture was filtered through a Buchner funnel, and the resulting filter cake was washed with deionized water. This product was dried under vacuum at room temperature to obtain the title compound (307 mg) as a yellow solid. Additional product was obtained from the filtrate. Ethyl acetate and THF were added to the filtrate to redissolve the material that had precipitated due to the reduction of the solvent under vacuum. The organic phase was washed with brine and dried over magnesium sulfate. The drying agent was filtered off and the solvents were removed in vacuo using a rotary evaporator to obtain an additional amount (492 mg) of product. ¹ H NMR (DMSO-d ₆ ) δ 7.94-8.00 (m, 1H), 7.89-7.94 (m, 1H), 3.74-3.84 (m, J=4.4 Hz, 4H), 3.54-3.62 (m, J= 4.3 Hz, 4H), 3.52 (s, 3H), 1.44 (s, 9H).

Intermediate 21: Methyl 8-(4-(tert-butoxycarbonyl)piperazin-1-yl)-7-cyano-5methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carboxylate

Palladium (II) acetate (1.502 mg, 6.69 µmol), 1,1'-bis(diphenylphosphino)ferrocene (7.42 mg, 0.013 mmol) and tert-butyl were added to a two-dram test tube equipped with a diaphragm cap. 4-(6bromo-3-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)piperazine-1-carboxylate (30 mg, 0.067 mmol). DMF (1.1 ml) was added to the tube (sparged with nitrogen for 30 min). Test tube

- 35 044500 was degassed and then filled with carbon monoxide from a cylinder (10x) through a needle and 3-way valve, then methanol (0.46 ml) was added. This reaction mixture was heated for 1 hour at 60°C. Triethylamine (0.020 mL, 0.143 mmol) was then added. Heating of the reaction mixture at 60°C in an atmosphere of carbon monoxide (1 at.) was continued overnight. The reaction mixture was then cooled and filtered through a syringe filter. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 20-60% B for 25 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 98.9%; measured mass: 428.11; retention time: 1.65 min. Analysis conditions for the 2nd sample injection:

column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 428.13; retention time: 1.69 min. Proton NMR signal intensities near the frequency used to suppress the water signal may be distorted and were not corrected: ¹ H NMR (DMSO-d ₆ ) δ 8.25 (d, J=8.8 Hz, 1H), 8.09 (d, J=8.8 Hz , 1H), 3.93 (s, 3H), 3.85-3.92 (m, J=4.8 Hz, 4H), 3.60-3.70 (m, J=3.7 Hz, 4H), 3.56 (s, 3H), 1.45 (s, 9H).

Intermediate 22: Methyl 7-cyano-5-methyl-6-oxo-8-(piperazin-1-yl)-5,6-dihydro-1,5naphthyridine-2-carboxylate, HCl

Into a 2 dram test tube containing methyl 8-(4-(tert-butoxycarbonyl)piperazin-1-yl)-7cyano-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carboxylate (109 mg, 0.255 mmol) dioxane (1.5 ml) was added. This mixture was heated to dissolve the solids. The mixture was then cooled, which led to the formation of a precipitate, and then 3 ml of 4N was added. solution of HCl in dioxane. The tube was capped and the reaction mixture was stirred for 1.45 hours at room temperature. Monitoring the reaction using HPLC indicated that the reaction was complete. The reaction mixture was transferred to a round bottom flask. Volatile components were removed under vacuum using a rotary evaporator. The resulting material was dried under vacuum at room temperature to yield 94 mg of product as a light pink solid. LC-MS: column: Waters Acquity VEN 2.1 mm x 50 mm, particle size 1.7 µm; mobile phase A: (10:90) acetonitrile:water with 0.1% TFA; mobile phase B: (90:10) acetonitrile:water with 0.1% TFA; temperature: 40°C; gradient: from 0% B to 100% B for 2 min, then maintaining 100% B for 1 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Injected sample volume: 1 µl. Retention time = 0.812 min; observed adducts: [M+H]; measured masses: 328.2. 1H NMR (DMSO-dg) δ 9.22-9.43 (m, 2H), 8.30 (d, J=9.0 Hz, 1H), 8.16 (d, J=9.0 Hz, 1H), 4.01-4.15 (m, 4H), 3.93 (s, 3H), 3.58 (s, 3H), 3.42 (br. s., 4H).

Intermediate 23: tert-butyl 4-(3-cyano-6-(methoxy(methyl)carbamoyl)-1-methyl-2oxo-1,2-dihydro-1,5-naphthyridin-4-yl)piperazine-1 - carboxylate

Palladium (II) acetate (1.5 mg, 6.68 µmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (3.9 mg, 6 .74 µmol), tert-butyl 4-(6-bromo-3-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)piperazin-1-carboxylate (30

- 36 044500 mg, 0.067 mmol) and Ν,Ο-dimethylhydroxylamine hydrochloride (10.2 mg, 0.105 mmol). DMF (1.0 ml) was added (sparged with nitrogen for 30 min) and the tube was degassed and then filled with carbon monoxide from a cylinder (1x) through the needle and 3-way valve. Triethylamine (30 μL, 0.215 mmol) was then added to the reaction mixture. The reaction mixture was heated in an oil bath (60°C) and stirred under a carbon monoxide atmosphere (1 atm; balloon pressure) for 18 hours. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 microns; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 20-60% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 457.12; retention time: 1.48 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 457.15; retention time: 1.59 min. Proton NMR signal intensities near the frequency used to suppress the water signal are distorted and uncorrected in this analysis. 1H NMR (DMSO) δ 8.08 (d, J=8.8 Hz, 1H), 7.90 (d, J=8.4 Hz, 1H), 3.79-3.88 (m, J=4.8 Hz, 4H), 3.61 (s, 2H) , 3.53-3.59 (m, 6H), 3.31 (s, 2H), 1.45 (s, 9H).

Intermediate 24: 8-(4-(tert-butoxycarbonyl)piperazin-1-yl)-7-cyano-5-methyl-6oxo-5,6-dihydro-1,5-naphthyridine-2-carboxylic acid

Intermediate 24 was isolated as a reaction by-product to give the previous product, tert-butyl 4-(3-cyano-6-(methoxy(methyl)carbamoyl)-1-methyl-2-oxo-1,2dihydro-1,5- naphthyridin-4-yl)piperazine-1-carboxylate. This crude substance was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 20-60% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; RT: 1.5; observed adducts: [M+Na]; measured masses: 436.05. Proton NMR signal intensities near the frequency used to suppress the water signal are distorted and uncorrected. 1H NMR (500 MHz, DMSO-d ₆ ) δ 8.21 (d, J=8.8 Hz, 1H), 8.04 (d, J=9.2 Hz, 1H), 3.87 (br.s, 4H), 3.62 (br.s , 2H), 3.54 (s, 1H), 1.43 (s, 9H).

Intermediate 25: Ethyl 3-acetamidopicolinate

O^CHZ

[^γΝΗ o (1-25)

Ethyl 3-aminopicolinate (5.0 g, 30.1 mmol) was dissolved in THF (35 ml) with heating. Acetic anhydride (13 mL, 138 mmol) was then added to the reaction mixture. The reaction mixture was placed under a nitrogen atmosphere and heated to reflux for 3 hours. The reaction mixture was then cooled

- 37 044500 and volatile components were removed under vacuum using a rotary evaporator. The resulting colorless product was dried under vacuum at room temperature to yield the title compound (6.228 g). LC-MS: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: 100% water with 0.05% trifluoroacetic acid; mobile phase B: 100% acetonitrile with 0.05% trifluoroacetic acid; temperature: 40°C; gradient: 2-98% B for 1.5 min, then maintain 98% B for 0.5 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. Retention time = 0.85 min; observed adducts: [M+H]; measured masses: 209.1. 1H NMR (DMSO-d ₆ ) δ0.35 (br.s., 1H), 8.33-8.40 (m, 2H), 7.59 (dd, J=8.4, 4.5 Hz, 1H), 4.32 (q, J= 7.1 Hz, 2H), 2.11 (s, 3H), 1.32 (t, J=7.2 Hz, 3H).

Intermediate 26: Ethyl 3-(N-methylacetamido)picolinate

Ethyl 3-acetamidopicolinate (1.0 g, 4.80 mmol) was dissolved in DMF (48.0 ml). Cesium carbonate (2.191 g, 6.72 mmol) was added and then methyl iodide (0.480 mL, 7.68 mmol) was added. The test tube with the reaction mixture was capped and the reaction mixture was stirred for 2.25 hours at room temperature. Volatile components were removed under vacuum using a system consisting of a rotary evaporator and a vacuum pump. The residue was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate. The aqueous phase was extracted with ethyl acetate. The organic extracts were combined and washed successively with saturated aqueous sodium bicarbonate followed by brine. The organic extract was dried over magnesium sulfate, filtered and the solvent was removed under vacuum using a rotary evaporator. The title compound was obtained as a light brown solid (891 mg). LC-MS: Column: Waters Acquity

UPLC VEN C18, 2.1x50 mm, particle size 1.7 microns; mobile phase A: 100% water with 0.05% trifluoroacetic acid; mobile phase B: 100% acetonitrile with 0.05% trifluoroacetic acid; temperature: 40°C; gradient: 2-98% B for 1.5 min, then maintain 98% B for 0.5 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. Retention time = 0.80 min; observed adducts: [M+H]; measured masses: 223.1. 1H NMR (acetonitrile-d ₃ ) δ 8.47-8.70 (m, 1H), 7.66-7.88 (m, 1H), 7.50-7.65 (m, 1H), 4.24-4.41 (m, 2H), 3.04-3.38 (m , 3H), 1.69 (s, 2H), 1.32 (t, J=7.0 Hz, 3H).

Intermediate 27: 4-hydroxy-1-methyl-1,5-naphthyridin-2(1H)-one

A solution of KHMDS (4.1 mL, 4.10 mmol) in THF (1.0 M) was added to a 100 mL round bottom flask under a nitrogen atmosphere and cooled to -78°C. A solution of ethyl 3-(Nmethylacetamido)picolinate (805 mg, 3.62 mmol) in THF (36.5 ml) was then added over 26 min. This reaction mixture was stirred at -78°C for 1 hour. The flask was removed from the dry ice bath. The reaction mixture was warmed to room temperature with stirring for 1.5 hours. Ethyl acetate and 30 ml of deionized water were added to the reaction mixture, then the reaction mixture was transferred to a separatory funnel and shaken gently. The aqueous phase was separated and transferred to an Erlenmeyer flask, acidified by adding 1N. hydrochloric acid solution (4.1 ml, 4.10 mmol) and stirred at room temperature. The resulting precipitate was collected by filtration using a Buchner funnel. The pH of the filtrate, measured using a pH test strip, was approximately 4. The resulting colorless precipitate was dried under vacuum at room temperature to yield 531 mg of the title compound as a colorless solid. LC-MS: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: 100% water with 0.05% trifluoroacetic acid; mobile phase B: 100% acetonitrile with 0.05% trifluoroacetic acid; temperature: 40°C; gradient: 2-98% B for 1.5 min, then maintain 98% B for 0.5 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. Retention time = 0.56 min; observed adducts: [M+H]; measured masses: 177.0. 1H NMR (DMSO-d ₆ ) δ 10.97 (br. s., 1H), 8.51 (dd, J=4.3, 1.0 Hz, 1H), 7.96 (dd, J=8.5, 1.0 Hz, 1H), 7.68 (dd , J=8.5, 4.5 Hz, 1H), 6.06 (s, 1H), 3.53 (s, 3H).

Intermediate 28: 4-chloro-1-methyl-1,5-naphthyridin-2(1H)-one

- 38 044500

A suspension of 4-hydroxy-1-methyl-1,5-naphthyridin-2(1H)-one (100 mg, 0.568 mmol) in acetonitrile (2.8 ml) was obtained. Phosphorus oxychloride (0.423 mL, 4.54 mmol) was added to this suspension. The reaction vessel was capped under a nitrogen atmosphere and heated at 80°C for 4 hours. The reaction mixture was then cooled and analyzed using HPLC. Additional phosphorus oxychloride (0.2 mL, 2.146 mmol) was added to the reaction mixture and the reaction vessel was capped under nitrogen and heated to 80°C for 2 hours. The reaction mixture was then cooled and volatiles were removed under vacuum using a rotary evaporator. . The remainder of the reaction mixture was partitioned between ethyl acetate and 1.5 M aqueous K2HPO4 solution. The aqueous phase (pH ~7, as indicated by the pH strip) was made alkaline by adding saturated aqueous sodium bicarbonate and then extracted with ethyl acetate. The organic extracts were combined and washed successively with saturated aqueous sodium bicarbonate and brine. The organic extract was then dried over magnesium sulfate, filtered and the solvent was removed under vacuum to obtain 138 mg of product. LC-MS: column: Phenomenex LUNA C18, 2 mmx 50, particle size 3 µm; mobile phase A: 5% acetonitrile: 95% water: 10 mM ammonium acetate; mobile phase B: 95% acetonitrile: 5% water: 10 mM ammonium acetate; temperature: 40°C; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. Retention time = 1.69 min; observed adducts: [M+H]; measured masses: 194.9. ¹ H NMR (DMSO) δ 8.64 (dd, J=4.4, 1.3 Hz, 1H), 8.10 (dd, J=8.7, 1.3 Hz, 1H), 7.76 (dd, J=8.6, 4.3 Hz, 1H), 7.23 (s, 1H), 3.63 (s, 3H).

Intermediate 29: 3-bromo-4-hydroxy-1-methyl-1,5-naphthyridin-2(1H)-one

HE (1-29)

4-Hydroxy-1-methyl-1,5-naphthyridin-2(1H)-one (500 mg, 2.84 mmol) was dissolved in DMF (8 ml) and then NBS (535 mg, 3.01 mmol) was added. The reaction vessel was capped under a nitrogen atmosphere. The reaction mixture was stirred for 2 hours at room temperature. The solvent was removed under vacuum using a rotary evaporator and 8 ml of deionized water was added to the crude reaction product. This suspension was stirred by rotating the reaction vessel and treated with ultrasound (pulses lasting 5-10 seconds), then filtered through a Buchner funnel and washed with deionized water (three portions of 8-10 ml). The resulting colorless product was dried under vacuum to yield 681 mg of the title compound as a colorless solid. LC-MS: column: Waters Acquity VEH C18 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: 100% water with 0.05% TFA; mobile phase B: 100% acetonitrile with 0.05% TFA; temperature: 40°C; gradient: from 2% B to 98% B for 1.5 min, then maintaining 98% B for 0.5 min; flow rate: 0.8 ml/min; registration: MS and UV (220 nm). Injected sample volume: 3 µl. Results: retention time = 0.64 min; observed adducts: [M+H]; measured masses: 254.9. ¹ H NMR (DMSO-d ₆ ) δ 11.86 (br.s., 1H), 8.57 (d, J=3.5 Hz, 1H), 8.06 (d, J=8.5 Hz, 1H), 7.76 (dd, J= 8.5, 4.4 Hz, 1H), 3.65 (s, 3H).

Intermediate 30: 3-bromo-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl trifluoromethanesulfonate

CH ₃

O_CF ₃ o ⁰ (1-30)

3-Bromo-4-hydrokCi-1-methyl-1,5-naphthyridin-2(1H)-one (200 mg, 0.784 mmol) was dissolved in acetonitrile (5 ml) containing DIEA (0.548 ml, 3.14 mmol) , at room temperature. This reaction mixture was placed under a nitrogen atmosphere and cooled to 0°C. Trifluoromethanesulfonic anhydride (0.265 mL, 1.568 mmol) in acetonitrile (2 mL) was slowly added dropwise to the reaction mixture using a syringe. The reaction mixture was stirred for 30 min at 0°C, then warmed to room temperature over 40 min and stirred for 30 min at room temperature. Volatiles were removed from the reaction mixture using a rotary evaporator and then the crude reaction mixture was partitioned between ethyl acetate and 1.5 M potassium hydrogen orthophosphate solution. The aqueous phase was extracted with ethyl acetate and the organic extracts were combined and washed successively with 1.5 M potassium hydrogen phosphate and saline. The organic extract was then dried over magnesium sulfate, then filtered and the filtrate devolatised using a rotary evaporator to obtain the crude product as a dark brown amorphous solid. The purity of this crude product was approximately 78%, as was

- 39 044500 was established by LC-MS analysis with registration at a UV wavelength of 220 nm. The resulting crude product was used without further purification. LC-MS: column: Waters Acquity VEH C18 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: 100% water with 0.05% TFA; mobile phase B: 100% acetonitrile with 0.05% TFA; temperature: 40°C; gradient: from 2% B to 98% B for 1.5 min, then maintaining 98% B for 0.5 min; flow rate: 0.8 ml/min; registration: MS and UV (220 nm). Injected sample volume: 3 µl. Results: retention time = 1.01 min; observed adducts: [M+H]; measured masses: 386.9.

Intermediate 31: tert-butyl yl)piperazine-1-carboxylate

4-(1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-4-

4-Chloro-1-methyl-1,5-naphthyridin-2(1H)-one (15 mg,

0.077 mmol) was dissolved in DMF (0.7 ml) to obtain a solution. 1-BOC-piperazine (27.7 mg, 0.149 mmol) was then added and the resulting mixture was stirred until 1-BOC-piperazine dissolved, then potassium carbonate (19.2 mg, 0.139 mmol) was added. The reaction vessel was capped under a nitrogen atmosphere. The reaction mixture was heated for 16.5 hours at 85°C. LC-MS analysis of the reaction mixture showed 45% conversion of the starting materials into the reaction product. The reaction mixture was again placed under a nitrogen atmosphere and heated to 85°C for 71 hours. The reaction mixture was then diluted with acetonitrile (1 ml) and filtered through a syringe filter. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 25-65% B for 15 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. (Purity: 99.1%; RT: 1.61; observed adducts: [M+H]; measured masses: 345.18). Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. (Purity: 99.2%; RT: 1.48; observed adducts: [M+H]; measured masses: 345.18). Proton NMR signal intensities near the frequency used to suppress the water signal are distorted and uncorrected. 1H NMR (DMSO-d ₆ ) δ 8.49 (dd, J=4.2, 1.3 Hz, 1H), 7.86-8.04 (m, 1H), 7.61 (dd, J=8.6, 4.2 Hz, 1H), 6.04 (s, 1H), 3.53 (s, 4H), 1.42 (s, 9H).

Naphthyridine-2(1H)-oh intermediate

32: 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-1-methyl-1,5-

4-Chloro-1-methyl-1,5-naphthyridin-2(1H)-one (12.3 mg, 0.063 mmol) was dissolved in DMF (632 μl) to obtain a solution. 1-(4,4'-difluorobenzhydryl)piperazine (22.9 mg, 0.079 mmol) and potassium carbonate (18.7 mg, 0.135 mmol) were then added. This reaction mixture was placed under a nitrogen atmosphere and heated for 18 hours at 80°C. HPLC analysis showed approximately 50% conversion of the starting compounds

- 40 044500 substances into the reaction product. Potassium carbonate (8.2 mg, 0.059 mmol) was added to the reaction mixture. The reaction mixture was sealed under a nitrogen atmosphere and heated for 19 hours at 85°C. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 5090% B for 15 min, then maintain 100% B for 5 min; flow rate: 20 ml/min.

Fractions containing the desired product were combined and dried using a centrifugal evaporator. The product yield was 21.1 mg and its purity, determined by LC-MS analysis, was 100%. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. LC-MS: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Retention time = 1.42 min; observed adducts: [M+H]; measured masses: 447.1. LC-MS: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Retention time = 2.25 min; observed adducts: [M+H]; measured masses: 447.1. Proton NMR signal intensities near the frequency used to suppress the water signal are distorted and uncorrected 1H NMR (DMSOD6) δ 8.42 (d, J=2.9 Hz, 1H), 7.91 (d, J=8.8 Hz, 1H), 7.57 (dd , J=8.8, 4.4 Hz, 1H), 7.48 (dd, J=8.6, 5.7 Hz, 4H), 7.14 (t, J=8.8 Hz, 4H), 5.98 (s, 1H), 4.47 (s, 1H) , 3.51 (s, 2H).

Intermediate 33: Ethyl 6-bromo-3-(N-methylacetamido)picolinate

Ethyl 3-acetamido-6-bromopicolinate (5 g, 17.41 mmol) was dissolved in DMF (100 ml) to form a solution. Cesium carbonate (8.15 g, 25.01 mmol) and methyl iodide (1.75 mL, 28.0 mmol) were then added. This reaction mixture was placed under a nitrogen atmosphere and stirred for 2 hours and 40 minutes at room temperature. The solvent was removed under vacuum using a system consisting of a rotary evaporator and a vacuum pump. Ethyl acetate and DCM were added to the remainder of the reaction mixture along with chloroform and toluene. This mixture was filtered through a pad of diatomaceous earth (Celite) to remove salts. Solvents were again removed under vacuum using a rotary evaporator. The remainder of the reaction mixture was redissolved in chloroform and toluene and filtered through a pad of diatomaceous earth (Celite) to remove trace amounts of undissolved substances still present. Removal of the solvents in vacuo gave 5.35 g of the desired product as an orange oil. LC-MS: column: Waters Acquity VEH 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: 10:90 acetonitrile:water with 0.1% TFA; mobile phase B: 90:10 acetonitrile:water with 0.1% TFA; temperature: 40°C; gradient: from 0% B to 100% B for 2 min, then maintaining 100% B for 1 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Injected sample volume: 1 µl. Retention time = 1.07 min; observed adducts: [M+H]; measured masses: 301.1. The proton NMR spectrum showed characteristic features indicating limited rotation around a chemical bond (rotamer); 1H NMR (500 MHz, chloroform-d) δ 7.72 (d, J=8.4 Hz, 0.8H), 7.66 (d, J=8.4 Hz, 0.2H), 7.51 (d, J=8.4 Hz, 0.8H), 7.45 (d, J=8.4 Hz, 0.2H), 4.50-4.36 (m, 2.0H), 3.37 (s, 0.6H), 3.19 (s, 2.4H), 2.24 (s, 0.6H), 1.82 (s , 2.5H), 1.43-1.36 (m, 3.1H).

Intermediate 34: Ethyl 6-methyl-3-(N-methylacetamido)picolinate

- 41 044500

Ethyl 6-bromo-3-(N-methylacetamido)picolinate (3.01 g, 10.00 mmol) was dissolved in THF (100 ml).

This solution was bubbled with nitrogen for 20 min and then tetrakis(triphenylphosphine)palladium(0) (235 mg, 0.203 mmol) was added. The headspace of the reaction mixture was purged with nitrogen and dimethylzinc (12 mL, 12.00 mmol) was added via syringe. This reaction mixture was heated under nitrogen atmosphere for 2 hours at 70°C. The reaction mixture was then cooled and transferred to a 500 mL Erlenmeyer flask containing a magnetic stir bar. Ethyl acetate was added and saturated sodium bicarbonate solution was added slowly with stirring. The organic phase was separated from the aqueous phase, then washed with brine and dried over magnesium sulfate. The drying agent was separated by filtration from the organic extract and the solvent was removed under vacuum using a rotary evaporator to obtain 2.23 g of product as a yellow oil. This crude product was purified on an Isco Redi-sep silica gel cartridge (80 g) using a gradient of 20%-50% ethyl acetate in dichloromethane for elution. Fractions containing the desired product by TLC analysis were combined and the solvent was removed in vacuo to give the title compound (1.937 g) as a pale yellow oil. The purity of this product was approximately 92% by HPLC analysis detected at a UV wavelength of 220 nm. The analytically pure sample was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 5-45% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 237.17; retention time: 1.05 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 237.18; retention time: 1.01 min. ¹ H NMR (400 MHz, DMSO-06) δ 7.87 (d, J=8.1 Hz, 0.8H), 7.68 (d, J=8.1 Hz, 0.2H), 7.56 (d, J=8.1 Hz, 0.8H) , 7.49 (d, J=8.3 Hz, 0.2H), 4.31 (q, J=7.1 Hz, 1.6H), 4.24 (q, J=7.1 Hz, 0.4H), 3.29 (s, 0.6H), 3.04 ( s, 2.4H), 2.55 (s, 2.4H), 2.12 (s, 0.6H), 1.66 (s, 2.4H), 1.27 (t, J=7.1 Hz, 3.0H).

Intermediate 35: Ethyl 6-methyl-3-(methylamino)picolinate

To a solution of ethyl 6-methyl-3-(U-methylacetamido)picolinate (1 g, 3.68 mmol) in ethanol (20 mL) was added HCl (1.841 mL, 22.09 mmol). This reaction mixture was stirred for 6 hours at 80°C. Solvents were removed in vacuo and the resulting residue was dissolved in water (10 ml), made basic by adding 10% NaHCO ₃ (20 ml) and extracted with DCM (3x20 ml). The combined organic layers were washed with 10% NaHCO ₃ (20 ml), water (20 ml) and brine (20 ml). The organic fraction was dried over sodium sulfate and concentrated to give the crude product (0.8 g). This crude product was purified by silica gel column chromatography (12 g) using 40% ethyl acetate in petroleum ether to isolate the desired product (0.5 g, 2.55 mmol, yield: 69.2%). 1H NMR (acetonitrile^) δ 7.38 (br. s, 1H), 7.23 (d, J=8.7 Hz, 1H), 7.11 (d, J=8.7 Hz, 1H), 4.32 (q, J=7.1 Hz, 2H ), 2.87 (d, J=5.2 Hz, 3H), 2.36 (s, 3H), 1.34 (t, J=7.1 Hz, 3H).

Intermediate 36: 1,6-dimethyl-2,4-dioxo-1,2,3,4-tetrahydro-1,5-naphthyridine-3carbonitrile

- 42 044500

To a solution of ethyl 6-methyl-3-(methylamino)picolinate (3.0 g, 15.45 mmol), 2-cyanoacetic acid (1.445 g, 16.99 mmol) and PYBROP (7.20 g, 15.45 mmol ) in dichloromethane (77 ml), triethylamine (5.40 ml, 30.9 mmol) was added. This reaction mixture was stirred overnight. The reaction mixture was then washed with saturated aqueous NaHCO3 (30 ml) and then brine (30 ml) and the organic layer was dried (sodium sulfate), filtered and evaporated to obtain the crude product. This crude product was dissolved in a minimal amount of dichloromethane and purified by flash silica gel chromatography. The chromatographic column was pre-equilibrated with 1% TEA/5% EA/94% hexanes (94:5:1 (v/v) hexane/ethyl acetate and 1% TEA) and eluted with a step gradient of 20-100% ethyl acetate/ hexane/1% TEA and then 0-20% (v/v) methanol in dichloromethane with 1% TEA gave the pure compound. Fractions containing pure product were combined, concentrated by rotary evaporation and dried under high vacuum to yield a pale yellow solid (6 g). The resulting solid was redissolved in DCM and washed with 1N. NaOH solution. After shaking this mixture, a white solid precipitated. This solid was isolated from the mixture by filtration and dried under vacuum to give 1,6-dimethyl-2,4-dioxo-1,2,3,4-tetrahydro-1,5-naphthyridine-3-carbonitrile (3.1 g , 14.40 mmol, yield: 93%). LC-MS: Method S1; RT 1.027, M ⁺¹ 216. 1H NMR (500 MHz, methanol-d4) δ 7.87-7.73 (m, 1H), 7.47 (d, J=8.7 Hz, 1H), 3.56 (s, 3H), 2.61 (s , 3H).

Intermediate 37: 4-chloro-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile

CH ₃ ί Y τ

H ₃ C Ν γΐΝ ci (1-37)

A suspension of 4-hydroxy-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (600 mg, 2.79 mmol) was dissolved in POCl ₃ (2.86 ml, 30 .7 mmol). This reaction mixture was stirred for 2 hours at 120°C. Then the reaction mixture was cooled to room temperature and poured into 1N. NaOH solution (20 ml), cooled to 0°C. The precipitated solid was collected by filtration, washed with diethyl ether and dried under vacuum overnight to give 4-chloro1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (400 mg, 1.712 mmol, yield: 61.4%). LC-MS: Method S1; RT 1.274, M ⁺¹ 262. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ 8.25-8.04 (m, 1H), 7.77 (d, J=8.7 Hz, 1H), 3.64 (s, 3H), 2.63 (s, 3H).

Intermediate 38: 4-hydroxy-1,6-dimethyl-1,5-naphthyridin-2(1H)-one

CH ₃

JT y τ

_H3CN ^^

HE (1-38)

To a 1.0 M solution of NaHMDS (9 ml, 9.00 mmol) in THF at -78°C under a nitrogen atmosphere, a solution of ethyl 6-methyl-3(N-methylacetamido) was added dropwise through a cannula over 50 min with stirring on a magnetic stirrer )picolinate (1.825 g, 7.51 mmol) in THF (60 ml). This reaction mixture was stirred for an additional 70 minutes at -78°C, then the cold bath was removed and the reaction mixture was warmed to room temperature over 30 minutes. Ethyl acetate and then 45 ml of deionized water were added to the reaction mixture. This mixture was transferred to a separatory funnel, shaken, and the aqueous layer was separated and transferred to an Erlenmeyer flask. The aqueous layer was acidified by adding 1.0N with stirring. hydrochloric acid solution (9.0 ml, 9.00 mmol). This aqueous solution was stirred for several minutes and an orange solid precipitated from the aqueous mixture (pH 4, measured using a pH test strip). This orange product was collected by filtration using a Buchner funnel and washed with very little deionized water. The resulting product was dried under vacuum at room temperature to yield 840 mg of the title compound as an orange solid.

LC-MS: column: Waters Acquity VEN 2.1 mm x 50 mm, particle size 1.7 µm; mobile phase A: (10:90) acetonitrile:water with 0.1% TFA; mobile phase B: (90:10) acetonitrile:water with 0.1% TFA; temperature: 40°C; gradient: from 0% B to 100% B for 2 min, then maintaining 100% B for 1 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Injected sample volume: 1 µl. Retention time =1.12 min; observed adducts: [M+H]; measured masses: 191.2. 1H NMR (DMSO-d6) δ 10.67 (br.s., 1H), 7.86 (d, J=8.7 Hz, 1H), 7.53 (d, J=8.8 Hz, 1H), 6.03 (s, 1H), 3.51 (s, 3H), 2.57 (s, 3H).

Intermediate 39: 4-chloro-1,6-dimethyl-1,5-naphthyridin-2(1H)-one

- 43 044500

4-Hydroxy-1,6-dimethyl-1,5-naphthyridin-2(1P)-one (54.5 mg, 0.287 mmol) was suspended in acetonitrile (1.3 ml) in a 2-dram high-pressure tube and equipped with a sealed lid. POCl ₃ (0.3 mL, 3.22 mmol) and a magnetic stir bar were then added. The tube was capped under a nitrogen atmosphere and the reaction mixture was heated for 3 hours at 80°C. LC-MS analysis of the reaction mixture showed approximately 40% conversion. The reaction mixture was heated under nitrogen at 80°C for an additional 2 hours. The reaction was stopped by adding ice and then ethyl acetate. The mixture was transferred to a separatory funnel, additional ethyl acetate was added and washed with 1.5 M potassium hydrogen orthophosphate solution. The aqueous layer was extracted with ethyl acetate. The organic extracts were combined and washed successively with a small amount of 1.5 M potassium hydrogen orthophosphate solution and saline. The resulting organic extract was dried over magnesium sulfate, then the drying agent was filtered off and the solvent was removed from the filtrate under vacuum using a rotary evaporator to obtain the title compound (51 mg) as a beige solid. LC-MS: column: Waters Acquity VEN 2.1 mm x 50 mm, particle size 1.7 µm; mobile phase A: (10:90) acetonitrile:water with 0.1% TFA; mobile phase B: (90:10) acetonitrile:water with 0.1% TFA; temperature: 40°C; gradient: from 0% B to 100% B for 2 min, then maintaining 100% B for 1 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Injected sample volume: 1 µl. Retention time = 0.99 min; observed adducts: [M+H]; measured masses: 209.3. 1H NMR (chloroform-d) δ 7.64 (d, J=8.7 Hz, 1H), 7.43 (d, J=8.7 Hz, 1H), 7.13 (s, 1H), 3.69 (s, 3H), 2.71 (s, 3H).

Intermediate 40: 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-1,6-dimethyl-1,5naphthyridin-2(1H)-oh

Into a 20 mL high pressure tube containing the 2nd generation RuPhos precatalyst (27.9 mg, 0.036 mmol), 4-chloro-1,6-dimethyl-1,5-naphthyridin-2(1P)-one ( 150 mg, 0.719 mmol), 1-(4,4'-difluorobenzhydryl)piperazine (249 mg, 0.863 mmol) and cesium carbonate (703 mg, 2.157 mmol), 7.1 ml of solvent (DMA/tert-ViOP; 1) was added :4). The reaction tube was capped under a nitrogen atmosphere and immersed in an oil bath at 70°C. The bath temperature was increased to 90°C. The reaction mixture was heated for 18 hours at 90°C. Volatile components were removed from the reaction mixture under vacuum using a system consisting of a rotary evaporator and a vacuum pump. Chloroform and dichloromethane were added to the reaction mixture and the mixture was heated with vortex stirring and then filtered through a Whatman autovial filter (0.45 μm pore size) using a diatomaceous earth pad (Celite) as a prefilter. The solvent was removed from the filtrate to give the crude product as an orange oil. This crude product was purified by silica gel column chromatography, eluting with 15% ethyl acetate in dichloromethane. Fractions containing pure product by TLC were combined and the solvent was removed in vacuo using a rotary evaporator to give 290 mg of product as an orange oil. Removal of the solvent in vacuo gave the product (217 mg) as a pale yellow foam. LC-MS: column: Waters Acquity VEN 2.1 mm x 50 mm, particle size 1.7 µm; mobile phase A: (10:90) acetonitrile:water with 0.1% TFA; mobile phase B: (90:10) acetonitrile:water with 0.1% TFA; temperature: 40°C; gradient: from 0% B to 100% B for 2 min, then maintaining 100% B for 1 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Injected sample volume: 1 µl. Retention time = 1.18 min; observed adducts: [M+H]; measured masses: 461.3. LC-MS: column: Phenomenex Luna C18, 2 mmx50 mm, particle size 3 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 40°C; gradient: from 0% B to 100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: MS and UV (220 nm). Injected sample volume: 3 µl. Retention time = 3.61 min; observed adducts: [M+H]; measured masses: 461.2. 1P NMR (Chloroform-d) δ 7.56 (d, J=8.7 Hz, 1P), 7.38-7.43 (m, 4P), 7.29 (d, J=8.7 Hz, 1P), 6.97- 44 044500

7.03 (m, 4H), 6.16 (s, 1H), 4.31 (s, 1H), 3.62 (s, 3H), 3.57 (br. s., 4H), 2.63 (t, J=4.8 Hz, 4H), 2.57 (s, 3H).

Intermediate 41: tert-butyl 4-(1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4yl)piperazine-1-carboxylate

In a 30 ml tube containing the 2nd generation precatalyst RuPhos (46.5 mg, 0.060 mmol), 1-BOC-piperazine (268 mg, 1.438 mmol) and 4-chloro-1,6-dimethyl-1,5- naphthyridin-2(1H)-one (250 mg, 1.198 mmol), cesium carbonate (1171 mg, 3.59 mmol) and a magnetic stir bar were added. To this reaction mixture was added 12 ml of tert-butanol/dimethylacetamide (4:1). The tube was capped under a nitrogen atmosphere and immersed in an oil bath (75°C). The bath temperature was increased to 90°C. The reaction mixture was heated for 19 hours at 90°C. The reaction mixture was then cooled and filtered through a pad of diatomaceous earth (Celite) to remove salts and washed with dichloromethane. The filtrate was devolatilized under vacuum using a rotary evaporator to obtain the crude product as an orange solid. The crude reaction mixture was purified by chromatography on silica gel, eluting with 70% ethyl acetate in dichloromethane. Fractions containing pure product by TLC were combined and the solvent was removed in vacuo using a rotary evaporator to obtain 304 mg of an orange solid. According to LC-MS analysis, the purity of the product after chromatographic purification was 70%. An analytically pure sample was obtained as a result of additional purification by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 0-65% B for 20 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Retention time = 1.69 min; observed adducts: [M+H]; measured masses: 359.2. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Retention time = 1.68 min; observed adducts: [M+H]; measured masses: 359.2. Proton NMR signal intensities near the frequency used to suppress the water signal are distorted and uncorrected: 1H NMR (DMSO^) δ 7.84 (d, J=8.8 Hz, 1H), 7.47 (d, J=8.4 Hz, 1H), 5.99 (s, 1H), 3.48-3.57 (m, 4H), 3.41-3.47 (m, 1H), 2.54 (s, 3H), 1.43 (s, 9H). The remainder of the mixture containing the reaction product was subjected to bromination and purified at this stage.

Intermediate 42. tert-butyl naphthyridin-4-yl)piperazine-1-carboxylate

4-(3-bromo-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-

tert-butyl 4-(1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)piperazine-1-carboxylate (575 mg, 1.364 mmol) was dissolved in DMF (12 ml ) until a solution is obtained. This reaction mixture was cooled to 0°C under nitrogen, NBS (270 mg, 1.517 mmol) was added and the reaction mixture was stirred for 2 hours at 0°C. A small amount of saturated sodium bicarbonate solution was added to the reaction mixture and the mixture was stirred for several minutes, then the solvent

- 45 044500 and volatile components were removed using a system consisting of a rotary evaporator and a vacuum pump. The remainder of the reaction mixture was distributed between ethyl acetate and 1.5 M potassium hydrogen orthophosphate solution. The organic extract was washed successively with 1.5 M potassium hydrogen orthophosphate solution and saline solution. The organic extract was then dried over magnesium sulfate, filtered and the solvent was removed from the filtrate using a rotary evaporator. This crude product was purified on a silica gel column, eluting with 15% ethyl acetate in dichloromethane. Fractions containing pure product by TLC analysis were combined and the solvents were removed in vacuo using a rotary evaporator to obtain 611 mg of the title compound as a pale yellow solid. LC-MS: column: Waters Acquity VEN 2.1 mm x 50 mm, particle size 1.7 µm; mobile phase A: (10:90) acetonitrile:water with 0.1% TFA; mobile phase B: (90:10) acetonitrile:water with 0.1% TFA; temperature: 40°C; gradient: from 0% B to 100% B for 1.5 min, then maintaining 100% B for 0.5 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Injected sample volume: 1 µl. Retention time = 1.44 min; observed adducts: [M+H]; measured masses: 437.2. 1H NMR (chloroform-d) δ 7.57 (d, J=8.7 Hz, 1H), 7.33 (d, J=8.7 Hz, 1H), 3.73 (s, 3H), 3.64-3.70 (m, 4H), 3.60 ( br s, 4H), 2.62 (s, 3H), 1.52 (s, 9H).

Intermediate 43: 3-bromo-1,6-dimethyl-4-(piperazin-1-yl)-1,5-naphthyridin-2(1H)-one, TFA

tert-butyl 4-(3-bromo-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)piperazine-1-carboxylate (572 mg, 1.308 mmol) was dissolved in dichloromethane (6 ml) and then TFA (6.00 ml) was added to obtain a solution. This reaction mixture was placed under a nitrogen atmosphere and stirred for 1.5 hours at room temperature. Volatile components were then removed from the reaction mixture using a rotary evaporator. The resulting reaction product was redissolved in DCM and devolatilized several times under vacuum to remove excess TFA. The reaction product (1.086 g) was obtained as an orange oil, the weight of which indicated the presence of 4.4 equivalents of TFA. LCMS: column: Phenomenex Luna C18, 2 mmx50 mm, particle size 3 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 40°C; gradient: from 0% B to 100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: MS and UV (220 im). Injected sample volume: 3 µl. Retention time = 1.88 min; observed adducts: [M+H]; measured masses: 337.0.

Intermediate 44: Ethyl 6-bromo-3-(2-cyanoacetamido)picolinate

Add ethyl 3-amino-6-bromopicolinate (1.0 g, 4.08 mmol) and cyanoacetic acid (0.417 g, 4.90 mmol) to a 25 mL round bottom flask, then add DCM (14 mL) and N,N -dimethylaniline (0.622 ml, 5.30 mmol). The flask was capped and cooled to 0°C, then EDC (1.172 g, 6.11 mmol) was added. This reaction mixture was stirred for 10 minutes at 0°C, then warmed to room temperature and stirred for 2.5 hours. The reaction mixture was transferred to a separatory funnel, diluted with dichloromethane and washed successively with 1.0N. hydrochloric acid solution (3x) and saline solution (1x). The organic extract was dried over sodium sulfate, filtered and the solvent was removed from the filtrate using a rotary evaporator to give the title compound (1.177 g) as a beige solid. LC-MS: column: Waters Acquity VEH C18 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: 100% water with 0.05% TFA; mobile phase B: 100% acetonitrile with 0.05% TFA; temperature: 40°C; gradient: from 2% B to 98% B for 1.5 min, then maintaining 98% B for 0.5 min; flow rate: 0.8 ml/min; registration: MS and UV (220 nm). Injected sample volume: 3 µl. Results: retention time = 0.95 min; observed adducts: [M+H]; measured masses: 312.0. ^1H NMR (500 MHz, DMSO-b6) δ 10.67 (s, 1H), 8.23 (d, J=8.7 Hz, 1H), 7.89 (d, J=8.7 Hz, 1H), 4.34 (q, J=7.1 Hz, 2H), 4.06 (s, 2H), 1.33(1, J=7.2 Hz, 3H).

Intermediate 45: 6-bromo-4-hydroxy-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile

- 46 044500

Ethyl 6-bromo-3-(2-cyanoacetamido)picolinate (1.091 g, 3.50 mmol) was dissolved in anhydrous tetrahydrofuran (35.0 ml) to form a solution. This reaction solution was slightly heterogeneous. A solution of KHMDS (3.7 mL, 3.70 mmol) in THF (1.0 M) was added to the reaction solution at room temperature. This reaction mixture was stirred for 1.5 hours at room temperature. KHMDS (0.2 mL, 0.200 mmol) was then added and the reaction mixture was stirred at room temperature for several minutes. Hydrochloric acid (4.2 ml, 4.20 mmol) was added to obtain a pH in the range of 7-8 according to the pH indicator strip and then 2 ml of 1N was added. HCl solution. This mixture was stirred by rotating the vessel and the volatile components were removed under vacuum using a system consisting of a rotary evaporator and a vacuum pump. The resulting crude solid was suspended in 20 ml of deionized water and cooled in an ice bath, then the precipitate was collected by filtration using a Buchner funnel. This substance was washed with cold water, which was slowly percolated through the filter cake. The resulting product was dried under vacuum at room temperature. A proton NMR spectrum in DMSO-d ₆ confirmed the desired product but showed the presence of ammonium chloride. The resulting substance was resuspended in 20 ml of 1N. hydrochloric acid solution and filtered through a Buchner funnel. The filter cake was washed with 1N. a solution of hydrochloric acid and water. This product was dried to obtain the title compound (773 mg) as a light beige solid. LC-MS: column: Waters Acquity VEN 2.1 mm x 50 mm, particle size 1.7 µm; mobile phase A: (10:90) acetonitrile:water with 0.1% TFA; mobile phase B: (90:10) acetonitrile:water with 0.1% TFA; temperature: 40°C; gradient: from 0% B to 100% B for 2 min, then maintaining 100% B for 1 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Injected sample volume: 1 µl. Retention time = 0.62 min; observed adducts: [M+H]; measured masses: 266.0. 1H NMR (500 MHz, DMSO-d ₆ ) δ 11.90 (br.s., 1H), 7.84 (d, J=8.7 Hz, 1H), 7.62 (d, J=8.8 Hz, 1H).

Intermediate 46: 6-bromo-4-hydroxy-2-oxo-1-(prop-2-yn-1-yl)-1,2-dihydro-1,5naphthyridine-3-carbonitrile

A solution prepared by dissolving 6-bromo-4-hydroxy-2-oxo-1,2-dihydro-1,5-naphthyridine-3carbonitrile (704 mg, 2.65 mmol) in DMF (24 ml) was cooled to 0° C in a nitrogen atmosphere. Sodium hydride (60% by weight in mineral oil) (270 mg, 6.75 mmol) was added at one time. This reaction mixture was placed under a nitrogen atmosphere and stirred for 10 minutes, then the ice bath was removed and the reaction mixture was allowed to warm to room temperature for 35 minutes. Propargyl bromide (80% in toluene) (0.737 ml, 6.62 mmol) was added and the reaction mixture was stirred for 1 hour at room temperature. Acetic acid (0.8 mL, 13.97 mmol) was added and volatiles were removed from the reaction mixture using a rotary evaporator. This crude reaction product was dried under vacuum at room temperature for approximately 2 hours to leave a brown oil. To the crude reaction residue was added 12 ml of 1N. hydrochloric acid solution and this mixture was stirred by rotating the vessel and briefly treated with ultrasound. The mixture was cooled in an ice bath and then the resulting product was collected by filtration using a Buchner funnel and washed with 1N. hydrochloric acid solution (12 ml). The product was then dried under vacuum at room temperature to obtain 879 mg of the title compound as a tan solid. A proton NMR spectrum confirmed the desired product. LC-MS: column: Waters Acquity VEN 2.1 mm x 50 mm, particle size 1.7 µm; mobile phase A: (10:90) acetonitrile:water with 0.1% TFA; mobile phase B: (90:10) acetonitrile:water with 0.1% TFA; temperature: 40°C; gradient: from 0% B to 100% B for 2 min, then maintaining 100% B for 1 min; flow rate: 1 ml/min; registration: MS and UV (220 im). Injected sample volume: 1 µl. Retention time = 0.91 min; observed adducts: [M+H]; measured masses: 304.0. LC-MS: column: Phenomenex Luna C18, 2 mmx50 mm, particle size 3 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 40°C; gradient: from 0% B to 100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: MS and UV (220 nm). Injected sample volume: 5 µl. Retention time = 1.48 min; observed adducts: [M+H]; measured masses: 303.8. ¹ H NMR (DMSO-d ₆ ) δ 7.94-8.02 (m, 2H), 5.00 (d, J=2.4 Hz, 2H), 3.33 (t, J=2.4 Hz, 1H).

- 47 044500

Intermediate 47: 6-bromo-4-chloro-2-oxo-1-(prop-2-yn-1-yl)-1,2-dihydro-1,5-naphthyridine-3-carbonitrile

6-Bromo-4-hydroxy-2-oxo-1-(prop-2-yn-1-yl)-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (874 mg, 2.87 mmol) was added to acetonitrile (29 ml) to obtain a suspension. DIEA (3.1 mL, 17.75 mmol) was then added and the reaction mixture was stirred for several minutes to ensure dissolution of the components. Phosphorus oxychloride (POCl ₃ ) (1.1 mL, 11.80 mmol) and benzyltriethylammonium chloride (790 mg, 3.47 mmol) were then added to the reaction mixture. The reaction mixture was capped under a nitrogen atmosphere and stirred for 19 hours at room temperature. Volatile components were removed from the reaction mixture under vacuum using a rotary evaporator. Ice was added to the remainder of the reaction mixture, then dichloromethane and the mixture was transferred to a separatory funnel. A 1.5 M aqueous solution of potassium hydrogen orthophosphate was added, the phases were separated and the organic extract was washed with a 1.5 M solution of potassium hydrogen orthophosphate. The aqueous phases were combined and extracted with dichloromethane. The combined organic extracts were washed successively with 1.5 M potassium hydrogen phosphate and saline. This organic extract was dried over sodium sulfate and the drying agent was filtered off. The filtrate was devolatilized under vacuum using a rotary evaporator to obtain a sticky brown solid. This crude product was purified by normal phase chromatography on silica gel, eluting with dichloromethane. Fractions containing pure product by TLC analysis were combined and the solvent was removed in vacuo using a rotary evaporator to give the title compound (652 mg) as a pale yellow solid. LC-MS: column: Waters Acquity VEN 2.1 mm x 50 mm, particle size 1.7 µm; mobile phase A: (10:90) acetonitrile:water with 0.1% TFA; mobile phase B: (90:10) acetonitrile:water with 0.1% TFA; temperature: 40°C; gradient: from 0% B to 100% B for 2 min, then maintaining 100% B for 1 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Injected sample volume: 1 µl. Retention time = 1.30 min; observed adducts: [M+H]; measured masses: 321.95. ¹ H NMR (chloroform-d) δ 7.76-7.89 (m, 2H), 5.08 (d, J=2.5 Hz, 2H), 2.39 (t, J=2.5 Hz, 1H).

Intermediate 48: tert-butyl 4-(6-bromo-3-cyano-2-oxo-1-(prop-2-yn-1-yl)-1,2dihydro-1,5-naphthyridin-4-yl) piperazine-1-carboxylate

6-Bromo-4-chloro-2-oxo-1-(prop-2-yn-1-yl)-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (502 mg, 1.556 mmol) was dissolved in DMF (15 ml) to obtain a solution. Then 1-BOC-piperazine (296 mg, 1.588 mmol) was added and then potassium carbonate (323 mg, 2.335 mmol) was added. This reaction mixture was stirred at room temperature under nitrogen for 2 hours. DMF was removed under vacuum using a rotary evaporator and vacuum pump system. The remainder of the reaction mixture was partitioned between ethyl acetate and water. THF was added to improve solubility. For better phase separation, a 1.5 M aqueous solution of potassium hydrogen orthophosphate was added. The organic extract was washed successively with 1.5 M potassium hydrogen orthophosphate solution and saline solution. The organic phase was dried over magnesium sulfate and filtered. The solvent was removed under vacuum using a rotary evaporator. The resulting product was dried under vacuum at room temperature to yield the title compound as a yellow-greenish solid. LC-MS: column: Phenomenex LUNA C18, 2 mmx50 mm, particle size 3 µm; mobile phase A: 5% acetonitrile: 95% water: 10 mM ammonium acetate; mobile phase B: 95% acetonitrile: 5% water: 10 mM ammonium acetate; temperature: 40°C; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. Retention time = 3.10 min; observed adducts: [M+H]; measured masses: 471.9. 1H NMR (chloroform-d) δ 7.64-7.77 (m, 2H), 5.01 (d, J=2.4 Hz, 2H), 3.93 (br. s., 4H), 3.6.

- 48 044500

Intermediate 49: 6-bromo-2-oxo-4-(piperazin-1-yl)-1-(prop-2-yn-1-yl)-1,2-dihydro1,5-naphthyridine-3-carbonitrile

tert-Butyl 4-(6-bromo-3-cyano-2-oxo-1-(prop-2-in-1-yl)-1,2-dihydro-1,5-naphthyridin-4-yl)piperazin- 1-carboxylate and tetrahydrofuran (25 mg, 0.052 mmol) were dissolved in DCM (260 μl) to obtain a solution. TFA (260 μl) was then added. This reaction mixture was placed under a nitrogen atmosphere and stirred for 1.5 hours at room temperature. Volatile components were removed from the reaction mixture under vacuum using a rotary evaporator. The crude reaction mixture was dissolved in DMF/acetonitrile. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 10-50% B for 15 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 371.98; retention time: 1.09 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 371.97; retention time: 1.04 min. Proton NMR signal intensities near the frequency used to suppress the water signal are distorted and uncorrected. 1H NMR (DMSO-d ₆ ) δ 7.85-8.05 (m, 2H), 4.99 (d, J=2.6 Hz, 2H), 3.73-3.84 (m, 4H), 3.17-3.26 (m, 1H), 2.90- 3.01 (m, 4H).

Intermediate 50: tert-butyl 4-(6-bromo-1-methyl-3-nitro-2-oxo-1,2-dihydro-1,5naphthyridin-4-yl)piperazin-1-carboxylate

Tert-butyl piperazin-1 was added to a solution of 6-bromo-4-chloro-1-methyl-3-nitro-1,5-naphthyridin-2(1H)-one (20 mg, 0.063 mmol) in DMF (1 ml). -carboxylate (11.70 mg, 0.063 mmol) and DIPEA (0.033 ml, 0.188 mmol). This reaction mixture was heated at 35°C overnight. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 40-80% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A:

- 49 044500 (5:95) acetonitrile: water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 1st sample injection: purity: 100%; measured mass: 468.0; retention time: 2.1 min. Analysis results for the 2nd sample injection: purity: 99.1%; measured mass: 468.0; retention time: 2.1 min. The title compound (11.7 mg) was isolated in 39.7% yield.

Intermediate 51: 6-bromo-1-methyl-3-nitro-4-(piperazin-1-yl)-1,5-naphthyridin-2(1H)one

To a solution of tert-butyl 4-(6-bromo-1-methyl-3-nitro-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)piperazine-1-carboxylate (10.43 mg , 0.022 mmol) in dichloromethane (2 ml), trifluoroacetic acid (1.716 μl, 0.022 mmol) was added and this solution was stirred for 2 hours at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 5-40% B for 15 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 1st sample injection: purity: 100%; measured mass: 367.9; retention time: 1.0 minute. Analysis results for the 2nd sample injection: purity: 98.8%; measured mass: 367.9; retention time: 0.9 min. The title compound (5.2 mg) was isolated in 64.2% yield.

Intermediate 52: (1R,5S)-tert-butyl 3-(6-chloro-3-cyano-1-methyl-2-oxo-1,2-dihydro1,5-naphthyridin-4-yl)-3,8 -diazabicyclo[3.2.1]octane-8-carboxylate

To a solution of 4,6-dichloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (164 mg, 0.645 mmol) and triethylamine (0.270 ml, 1.936 mmol) in DMF ( 2 ml) (1R,5S)-tert-butyl 3,8diazabicyclo[3.2.1]octane-8-carboxylate (137 mg, 0.645 mmol) was added and the resulting mixture was stirred overnight at room temperature. HPLC analysis showed the transformation of the starting substances into a reaction product with a mass corresponding to the mass of the product without the tert-butyl group. The crude reaction mixture was evaporated under reduced pressure and the residue was dissolved in dichloromethane and then adsorbed onto silica gel and subjected to flash chromatography using 100% ethyl acetate as the eluent and silica gel as the stationary phase. Homogeneous fractions were combined and evaporated in vacuo to obtain 200 mg of a yellow solid. Analytical LCMS conditions: injected sample volume 1:8: 3 µl; initial %B: 2; final % B: 98; gradient duration: 1.5 min; flow rate: 0.8 ml/min; wavelength: 220 nm; solvent pair: water/acetonitrile/TFA; solvent A: 100% water/0.05% TFA; solvent B: 100% acetonitrile/0.05% TFA; column: Waters Aquity VEN C18 2.1x50 mm, 1.7 µm MW1; thermostat temperature: 40; RT: 0.794

- 50 044500 min. (M+H-bos) ⁺ ; 329. Purity was 77%. 1H NMR (500 MHz, DMSO-d ₆ ) δ 8.11 (d, J=9.1 Hz, 1H), 7.84 (d, J=9.0 Hz, 1H), 4.25 (br. s., 2H), 4.12 (s, 2H), 3.61-3.47 (m, 4H), 2.22 (d, J=7.6 Hz, 2H), 1.88-1.80 (m, 2H), 1.46 (s, 9H). 10 mg of this substance was then purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; gradient: 35-75% B for 15 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 1st sample input: purity: 100.0%; 2.04 min; [M+Na]; 452.06. Analysis results for the 2nd sample injection: purity: 100.0%; 2.14 min; [M+Na]; 452.05.

Intermediate 53: tert-butyl 4-(6-bromo-3-cyano-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridin-4-yl)piperazine-1-carboxylate

To a solution of 6-bromo-4-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (100 mg, 0.335 mmol) and triethylamine (0.047 ml, 0.335 mmol) in DMF (5 ml) was added tert-butyl piperazine-1carboxylate (62.4 mg, 0.335 mmol). The resulting mixture was stirred at room temperature under nitrogen atmosphere overnight. The DMF was then removed under vacuum. The residue was dissolved in dichloromethane and the resulting solution was washed successively with water (1x) and then with saline. The organic layer was then dried over magnesium sulfate, filtered and evaporated under reduced pressure to obtain the desired product as a yellow-orange solid (125 mg, 79%). Analytical LC-MS conditions: injection sample volume: 3 µl; initial %B: 2; final % B: 98; gradient duration: 1.5 min; flow rate: 0.8 ml/min; wavelength: 220 nm; solvent pair: water/acetonitrile/TFA; solvent A: 100% water/0.05% TFA; solvent B: 100% acetonitrile/0.05% TFA; column: Waters Aquity VEN C18 2.1x50 mm, 1.7 µm MW1; oven temperature: 40. LC-MS results: 1.207 min; (M-tert-Bu) ⁺ ; 391.85. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.05-7.85 (m, 2H), 3.81 (br. s., 4H), 3.67-3.44 (m, 7H), 1.45 (s, 9H).

Intermediate 55: tert-butyl 4-(6-cyano-1-methyl-3-nitro-2-oxo-1,2-dihydro-1,5naphthyridin-4-yl)piperazin-1-carboxylate

Huenig's base ( 0.535 ml, 3.06 mmol) and then tert-butyl piperazine-1-carboxylate (209 mg, 1.122 mmol) was added. This reaction mixture was shaken for 2 hours at room temperature. According to LC-MS analysis, the reaction was complete. The solvent was removed under vacuum. The residue was purified on a silica gel column (40 g), eluting with hexanes: ethyl acetate (1:1). Fractions containing the title compound were combined to obtain a yellow solid (320 mg, yield: 76%). Analytical LC-MS conditions: column: Phenomenex LUNA C18, 50x2, particle size 3 µm; mobile phase A: 5% acetonitrile: 95% water: 51 044500 yes: 10 mM ammonium acetate; mobile phase B: 95% acetonitrile: 5% water: 10 mM ammonium acetate; temperature: 40°C; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. Analytical LC-MS results confirmed the title compound: 1.8 min, 475 (M+H), 473 (M-H).

Intermediate 56: 5-methyl-7-nitro-6-oxo-8-(piperazin-1-yl)-5,6-dihydro-1,5naphthyridine-2-carbonitrile

N

N (1-56)

To a solution of tert-butyl 4-(6-cyano-1-methyl-3-nitro-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)piperazine-1-carboxylate (320 mg, 0.772 mmol) in dichloromethane (4 ml), TFA (0.059 ml, 0.772 mmol) was added and the reaction mixture was stirred for 2 hours at room temperature. According to LC-MS analysis, the reaction was complete. The reaction mixture was concentrated in vacuo to give a yellow solid (240 mg, yield: 73%), which was used without further purification. Analytical LC-MS conditions: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: 100% water with 0.05% trifluoroacetic acid; mobile phase B: 100% acetonitrile with 0.05% trifluoroacetic acid; temperature: 40°C; gradient: 2-98% B for 1.5 min, then maintain 98% B for 0.5 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. Analytical LC-MS results confirmed the title compound: 1.0 min, 315 (M+H).

Intermediate 57: 6-bromo-1-methyl-2,4-dioxo-1,2,3,4-tetrahydro-1,5-naphthyridine-3carbonitrile

HE (1-57)

To a solution of 6-bromo-2,4-dioxo-1,2,3,4-tetrahydro-1,5-naphthyridine-3-carbonitrile (250 mg, 0.940 mmol) in DMF (5 ml), 60% hydride was added in portions sodium (113 mg, 2.82 mmol) in mineral oil. This reaction mixture was stirred for 30 minutes at room temperature. Iodomethane (0.176 mL, 2.82 mmol) was added and the reaction mixture was stirred overnight at room temperature. LC-MS analysis indicated the presence of unreacted starting material. One equivalent of sodium hydride and iodomethane was added. The reaction mixture was stirred for 4 hours at room temperature. The reaction was then stopped by adding water. The reaction mixture was acidified to pH~3 by adding 1N. hydrochloric acid solution. A yellowish solid was collected to obtain the final reaction product (163 mg, yield: 61.9%). Analytical LCMS conditions: column: Phenomenex LUNA C18, 50x2, particle size 3 µm; mobile phase A: 5% acetonitrile: 95% water: 10 mM ammonium acetate; mobile phase B: 95% acetonitrile: 5% water: 10 mM ammonium acetate; temperature: 40°C; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. Analytical LC-MS results confirmed the title compound: 1.9 min, 280, 282 (M+H).

Intermediate 58: 6-methoxy-1-methyl-2,4-dioxo-1,2,3,4-tetrahydro-1,5-naphthyridine3-carbonitrile

HE (1-58)

6-Bromo-1-methyl-2,4-dioxo-1,2,3,4-tetrahydro-1,5-naphthyridine-3-carbonitrile (100 mg, 0.357 mmol) was mixed with sodium methoxide (0.5 N solution in methanol) (2.142 ml, 1.071 mmol) in 1 ml DMF and the mixture was heated in a microwave reactor for 5 hours at 85°C, then for 2 hours at 100°C. According to LC-MS analysis, the reaction was complete. The solid reaction product (40 mg, yield: 50%) was collected by filtration.

Intermediate 59: 4-chloro-6-methoxy-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3carbonitrile

- 52 044500

To a solution of 6-methoxy-1-methyl-2,4-dioxo-1,2,3,4-tetrahydro-1,5-naphthyridine-3-carbonitrile (70 mg, 0.245 mmol) in acetonitrile (1.5 ml) DIPEA (0.427 mL, 2.447 mmol) was added and then phosphorus oxychloride (0.160 mL, 1.713 mmol) was added. This mixture was stirred for 2 hours at room temperature. LC-MS analysis showed the formation of a new peak, confirming the completion of the reaction. Volatile components were removed under vacuum. The residue was redissolved in ethyl acetate, washed with water and brine, and dried over magnesium sulfate. The solvent was removed in vacuo to obtain a light yellow solid (40 mg, yield: 74%).

Intermediate 60: 4,6-dichloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3carbonitrile

CH ₃ dXX Cl NY^ CN

Cl (1-60)

To 6-bromo-4-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (1 g, 3.35 mmol) was added 4 M HCl solution (20 ml, 80 mmol) in dioxane. This reaction mixture was heated in hermetically sealed tubes at 85°C for 4 days. The reaction mixture was then concentrated and triturated with methanol. The solid was collected by filtration to obtain the hydrochloride salt of the title compound as a yellow solid (810 mg, yield: 83%). Analytical LC-MS conditions: column: Phenomenex LUNA C18, 2.0x50 mm, particle size 3 µm; mobile phase A: (10:90) methanol:water with 0.1% TFA; mobile phase B: (90:10) methanol:water with 0.1% TFA; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: 2.9 min, 253.9 (M+H)+. 1H NMR (400 MHz, chloroform-d) δ 7.80-7.76 (m, 1H), 7.71-7.68 (m, 1H), 3.77 (s, 3H).

Intermediate 61: tert-butyl 4-(6-bromo-3-cyano-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridin-4-yl)piperazin-1-carboxylate

To a solution of 6-bromo-4-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (1.2 g, 4.02 mmol) in DMF (10 ml) , tert-butyl piperazine-1-carboxylate (0.824 g, 4.42 mmol) and triethylamine (1.681 mL, 12.06 mmol) were added. The reaction was stopped by adding 1N. HC1 solution. The precipitated yellow solid was collected by filtration to give the title compound (1.775 g, yield: 98%). Analytical LC-MS conditions: column: Phenomenex LUNA C18, 2.0x50 mm, particle size 3 µm; mobile phase A: (10:90) methanol:water with 0.1% TFA; mobile phase B: (90:10) methanol:water with 0.1% TFA; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: 4.0 min, 448.0 (M+H) ⁺ . 1H NMR (400 MHz, chloroform-d) δ 7.67 (d, J=9.0 Hz, 1H), 7.55 (d, J=8.8 Hz, 1H), 3.93 (br. s, 4H), 3.79-3.69 (m, 4H), 3.64 (s, 3H), 1.52 (s, 9H).

Intermediate 62: 6-bromo-1-methyl-2-oxo-4-(piperazin-1-yl)-1,2-dihydro-1,5naphthyridine-3-carbonitrile

To a solution of tert-butyl 4-(6-bromo-3-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)piperazine-1-carboxylate (700 mg, 1.561 mmol) in dichloromethane (5 ml), trifluoroacetic acid (3

- 53 044500 ml, 38.9 mmol). This reaction mixture was stirred for 2 hours at room temperature and then concentrated in vacuo to obtain the bis-TFA salt of the title compound as a brownish solid (858 mg, 1.489 mmol, yield: 95%). Analytical LCMS conditions: column: Phenomenex LUNA C18, 2.0x50 mm, particle size 3 µm; mobile phase A: (10:90) methanol:water with 0.1% TFA; mobile phase B: (90:10) methanol:water with 0.1% TFA; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: 1.7 min, 348.0 (MH ⁺ ). ¹ H NMR (400 MHz, methanol-d ₄ ) δ 7.99 (d, J=9.0 Hz, 1H), 7.88 (d, J=9.0 Hz, 1H), 4.27-4.07 (m, 4H), 3.67 (s, 3H), 3.59-3.46 (m, 4H).

Intermediate 63: 5-methyl-6-oxo-8-(piperazin-1-yl)-5,6-dihydro-1,5-naphthyridine2,7-dicarbonitrile

Tert-butyl 4-(6-bromo-3-cyano-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridin-4-yl)piperazin-1-carboxylate (100 mg, 0.223 mmol), zinc (2.92 mg, 0.045 mmol), zinc cyanide (15.72 mg, 0.134 mmol) and 1,1'-bis(diphenylphosphino)ferrocene-palladium(II) dichloride and dichloromethane complex (18.22 mg, 0.022 mmol). This vessel was sequentially degassed and purged with nitrogen three times. NMP (2 ml) was added and the mixture was heated for 1 hour at 75°C. Methanol was added, the resulting suspension was filtered and the filtrate was purified by preparative reverse phase HPLC using methanol-H ₂ O-TFA as eluent. Homogeneous fractions were combined and then concentrated in vacuo overnight to give a light yellow solid. This material was dissolved in dichloromethane (3 ml) and TFA (3 ml, 38.9 mmol) was added. This reaction mixture was stirred for 3 hours at room temperature and then evaporated under reduced pressure to obtain the bis-TFA salt of the title compound as a reddish solid (53 mg, yield: 45.5%). Analytical LC-MS conditions: Phenomenex LUNA C18, 50x2, 3 µm particle size; mobile phase A: 5% acetonitrile: 95% water: 10 mM ammonium acetate; mobile phase B: 95% acetonitrile: 5% water: 10 mM ammonium acetate; temperature: 40°C; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: 1.8 min, 295.1 (M+H)+. 1H NMR (500 MHz, DMSO-d ₆ ) δ 8.22 (d, J=8.8 Hz, 1H), 8.13 (d, J=9.2 Hz, 1H), 3.87-3.75 (m, 4H), 3.55 (s, 3H ), 3.08-2.92 (m, 4H).

Intermediate 64: naphthyridine-3-carbonitrile

6-chloro-1-methyl-2-oxo-4-(piperazin-1 -yl)-1,2-dihydro-1,5-

To a solution of 4,6-dichloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (115 mg, 0.453 mmol) in DMF (2 ml), add tert-butyl piperazine -1-carboxylate (84 mg, 0.453 mmol) and triethylamine (0.189 ml, 1.358 mmol). This reaction mixture was stirred for 2 hours at room temperature. The reaction was stopped by adding water. The beige solid that precipitated out of solution was collected by filtration. This substance was then dissolved in dichloromethane (5 ml) and a 2M solution of HCl in diethyl ether (2.263 ml, 4.53 mmol) was added. The resulting mixture was stirred at room temperature for 2 days and then concentrated under vacuum to obtain the tribasic hydrochloride salt of the title compound as a beige solid (133 mg, yield: 71.1%). Analytical LC-MS conditions: Column: Phenomenex LUNA C18,

2.0x50 mm, particle size 3 microns; mobile phase A: (10:90) methanol:water with 0.1% TFA; mobile phase B: (90:10) methanol:water with 0.1% TFA; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: 1.8 min, 304.0 (M+H) ⁺ . 1H NMR (400 MHz, DMSO-d ₆ ) δ 9.30 (br. s., 2H), 8.13 (d, J=9.0 Hz, 1H), 7.86 (d, J=9.0 Hz, 1H), 4.10-3.89 ( m, 4H), 3.57 (s, 3H), 3.36 (br. s., 4H).

Intermediate 65: 2,4-dioxo-1,2,3,4-tetrahydro-1,5-naphthyridine-3-carbonitrile

- 54 044500

2-Cyanoacetic acid was converted to the congeneric acid chloride by treating a solution of 2-cyanoacetic acid (1.388 g, 16.32 mmol) in dichloromethane (10 ml) containing a few drops of DMF with a mixture of 2 M oxalyl chloride in dichloromethane (10.83 ml, 21.66 mmol) . The resulting solution was stirred for 3 hours at room temperature. The mixture was then concentrated and the residue was kept under vacuum overnight. This material was dissolved in dichloromethane (10 ml) and added dropwise to a solution of ethyl 3-aminopicolinate (1.5 g, 9.03 mmol) in dichloromethane (10 ml) containing DIPEA (6.31 ml, 36.1 mmol) . This reaction mixture was stirred at room temperature over the weekend and then quenched by adding water. The resulting mixture was extracted with dichloromethane. Then the aqueous layer was acidified with 1N. HCl solution to pH~2, the resulting precipitate was collected by filtration to obtain the hydrochloride salt of the title compound as a brown solid (689 mg, yield: 40.8%). Analytical LC-MS conditions: column: Phenomenex LUNA C18, 2.0x50 mm, particle size 3 µm; mobile phase A: (10:90) methanol:water with 0.1% TFA; mobile phase B: (90:10) methanol:water with 0.1% TFA; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: 0.44 min, 188.1 (MH ⁺ ). Ή NMR (400 MHz, DMSO-d ₆ ) δ 11.56 (br. s, 1H), 8.51 (dd, J=4.9, 1.2 Hz, 1H), 7.96 (dd, J=8.4, 1.1 Hz, 1H), 7.85 (dd, J=8.6, 4.9 Hz, 1H).

Intermediate 66: 1-methyl-2,4-dioxo-1,2,3,4-tetrahydro-1,5-naphthyridine-3-carbonitrile

Sodium hydride (60%, 278 mg, 6.95 mmol) in mineral oil. The resulting mixture was stirred for 30 minutes at room temperature. Iodomethane (0.651 mL, 10.42 mmol) was then added and stirring was continued overnight. The reaction was then stopped by adding water. The pH of the mixture was adjusted to ~3 by adding 1N dropwise. HCl solution. The separated solid from the mixture was collected by filtration to obtain the hydrochloride salt of the title compound as a solid powder (403 mg, 57.7%).

Intermediate 67: 4-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile

1-Methyl-2,4-dioxo-1,2,3,4-tetrahydro-1,5-naphthyridine-3-carbonitrile (250 mg, 1.243 mmol) and phosphorus oxychloride (2 ml, 21.46 mmol) were heated in for 5 hours at 95°C in a hermetically sealed tube. The reaction mixture was then concentrated under reduced pressure, the residue was poured into ice water and then neutralized by adding solid NaHCO ₃ . The resulting mixture was extracted with dichloromethane (2x20 ml) and the organic layers were combined, dried over MgSO ₄ and filtered. The filtrates were concentrated in vacuo to give the title compound as a brown solid (105 mg, yield: 38.5%). Analytical LC-MS conditions: column: Phenomenex LUNA C18, 2.0x50 mm, particle size 3 µm; mobile phase A: (10:90) methanol:water with 0.1% TFA; mobile phase B: (90:10) methanol:water with 0.1% TFA; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LCMS results: 2.3 min, 220.0 (MH ⁺ ). ^1H NMR (400 MHz, chloroform-d) δ 8.80 (dd, J=4.4, 1.2 Hz, 1H), 7.83 (dd, J=8.8, 1.2 Hz, 1H), 7.72 (dd, J=8.6, 4.4 Hz , 1H), 3.78 (s, 3H).

Intermediate 71: carboxylate

6-bromo-4-hydroxy-2-oxo-1,2-dihydro-1,5-naphthyridine-3ethyl

To a solution of ethyl 3-amino-6-bromopicolinate (500 mg, 2.040 mmol) and DIPEA (1.069 ml, 6.12 mmol) in dichloromethane (10 ml), ethyl 3-chloro-3-oxopropanoate (0.313 ml, 2.448 mmol) at

- 55 044500

0°C. The reaction mixture was then warmed to room temperature and stirred for 1 hour. 21% (w/w) sodium ethoxide (0.914 mL, 2.448 mmol) in ethanol was then added dropwise and the reaction mixture was stirred for 3 hours at room temperature. The reaction was then stopped by adding water. The reaction mixture was acidified to pH ~4 using 1N. HCl solution. The precipitated solid was collected by filtration to obtain the title compound as a brown solid (400 mg, yield: 62.6%). Analytical LC-MS conditions: Phenomenex LUNA C18, 50x2, particle size 3 µm; mobile phase A: 5% acetonitrile: 95% water: 10 mM ammonium acetate; mobile phase B: 95% acetonitrile: 5% water: 10 mM ammonium acetate; temperature: 40°C; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: 1.5 min, 312.9 (M+H)+. 1H NMR (400 MHz, methanol-d ₄ ) δ 7.81 (d, J=8.8 Hz, 1H), 7.64 (d, J=8.8 Hz, 1H), 4.45 (d, J=7.1 Hz, 2H), 1.42 ( t, J=7.2 Hz, 3H).

Intermediate 72: Ethyl naphthyridine-3-carboxylate

6-bromo-4-hydroxy-1-methyl-2-oxo-1,2-dihydro-1,5-

To a solution of ethyl 6-bromo-4-hydroxy-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carboxylate (400 mg, 1.278 mmol) in DMF (5 ml) was added portionwise 60% hydride sodium (128 mg, 3.19 mmol) in mineral oil. This reaction mixture was stirred for 30 minutes at room temperature, then iodomethane (0.200 ml, 3.19 mmol) was added. This mixture was stirred at room temperature for an additional 1 hour. The reaction was stopped by adding water. The reaction mixture was acidified to pH ~3 by adding 1N. HCl solution. The resulting solution was extracted with ethyl acetate (2x40 ml) and the organic layers were combined, dried (MgSO ₄ ) and concentrated in vacuo to give an orange oil. This substance was purified by reverse phase preparative HPLC using CH ₃ OH-H ₂ O-TFA as the eluent. Homogeneous fractions were combined, neutralized with saturated NaHCO ₃ solution and concentrated under reduced pressure to remove acetonitrile. The resulting mixture was extracted with ethyl acetate (2x40 ml) and the organic layers were combined, dried (MgSO ₄ ) and concentrated in vacuo to give the title compound as a yellow solid (190 mg, yield: 45.5%). Analytical LC-MS conditions: column: Phenomenex LUNA C18, 2.0x50 mm, particle size 3 µm; mobile phase A: (10:90) methanol:water with 0.1% TFA; mobile phase B: (90:10) methanol:water with 0.1% TFA; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: 3.1 min, 326.8 (M+H) ⁺ . 1H NMR (400 MHz, chloroform-d) δ 7.78 (d, J=8.8 Hz, 1H), 7.60 (d, J=8.8 Hz, 1H), 4.54 (q, J=7.2 Hz, 2H), 3.68 (s , 3H), 1.49 (t, J=7.1 Hz, 3H).

Intermediate 73: tert-butyl 4-((1H-indol-4-yl)methyl)piperazine-1-carboxylate Boc c) (1-73)

To a solution of tert-butyl piperazine-1-carboxylate (2.89 g, 15.50 mmol) in DMF (10 mL) was added 1H-indole-4-carbaldehyde (1.5 g, 10.33 mmol). This reaction mixture was stirred for 1 hour at room temperature, and sodium cyanoborohydride (1.948 g, 31.0 mmol) was added and the mixture was stirred at room temperature for 7 days. Water was then added and the separated solid was collected by filtration. This crude product was fractionated by flash chromatography on silica gel using 35% ethyl acetate in hexanes as eluent. Homogeneous fractions were combined and evaporated to obtain the title compound as a white solid (405 mg, yield: 12.43%). Analytical LC-MS conditions: column: Phenomenex LUNA C18, 50x2, particle size 3 µm; mobile phase A: 5% acetonitrile: 95% water: 10 mM ammonium acetate; mobile phase B: 95% acetonitrile: 5% water: 10 mM ammonium acetate; temperature: 40°C; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: 2.9 min, 316 (M+H) ⁺ . 1H NMR (400 MHz, chloroform-d) δ 8.23 (br.s., 1H), 7.35 (d, J=8.1 Hz, 1H), 7.23 (br.s., 1H), 7.18 (t, J=7.7 Hz, 1H), 7.13-7.05 (m, 1H), 6.76 (br. s., 1H), 3.83 (s, 2H), 3.51-3.40 (m, 4H), 2.53-2.40 (m, 4H), 1.48 (s, 9H).

Intermediate 74: 1-ethyl-4-(piperαzin-1-ylmethyl)-1H-indole bis(2,2,2trifluoroacetate)

- 56 044500

To a solution of tert-butyl 4-((1H-indol-4-yl)methyl)piperazine-1-carboxylate (0.4 g, 1.268 mmol) in DMF (5 ml) at 0°C, sodium hydride (0.061 g) was added in portions , 1.522 mmol, 60% in mineral oil). This reaction mixture was then warmed to room temperature and stirred for 30 minutes. Ethyl iodide (0.205 mL, 2.54 mmol) was then added and the reaction mixture was stirred for 10 min at room temperature, then quenched by adding water. The resulting mixture was extracted with ethyl acetate (2x40 ml). The extracts were combined, dried (MgSO ₄ ), filtered and concentrated in vacuo to give an orange oil. This substance was fractionated by preparative HPLC using acetonitrile-water-TFA mixture as the eluent. Homogeneous fractions were combined and concentrated under reduced pressure to obtain a dark brown viscous oil. This substance was dissolved in dichloromethane (3 ml) and TFA (1 ml) was added. This reaction mixture was stirred for 2 hours at room temperature. The reaction mixture was then concentrated in vacuo to give a dark purple oil. This substance was fractionated by reverse phase preparative HPLC using acetonitrile-water-TFA as an eluent. Homogeneous fractions were combined and concentrated in vacuo to give the title compound as a violet viscous oil (373 mg, yield: 62.4%). Analytical LC-MS conditions: column: Phenomenex LUNA C18, 2.0x50 mm, particle size 3 µm; mobile phase A: (10:90) methanol:water with 0.1% TFA; mobile phase B: (90:10) methanol:water with 0.1% TFA; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: 1.9 min, 244 (M+H)+. 1H NMR (400 MHz, methanol-d ₄ ) δ 7.60 (d, J=7.8 Hz, 1H), 7.43 (d, J=1.0 Hz, 1H), 7.35-7.15 (m, 2H), 6.74 (d, J =2.7 Hz, 1H), 4.68 (s, 2H), 4.29 (q, J=7.1 Hz, 2H), 3.68-3.43 (m, 8H), 1.45 (t, J=7.2 Hz, 3H).

Intermediate 75: 2-(phenyl(piperazin-1-yl)methyl)phenol

Phenylboronic acid (1.309 g, 10.74 mmol), salicylaldehyde (1.311 g, 10.74 mmol) and tert-butyl piperazine-1-carboxylate (2 g, 10.74 mmol) were dissolved in DMF (12 ml) in a round bottom flask. This reaction mixture was heated at 120°C in an oil bath over the weekend. The reaction mixture was then diluted with ethyl acetate and washed successively with water and brine. The organic layer was then dried (MgSO ₄ ), filtered and concentrated in vacuo to give a yellow viscous oil. This oil was subjected to flash chromatography on silica gel using 20% ethyl acetate in hexanes as the eluent. Homogeneous fractions were combined and concentrated in vacuo to give the protected derivative as a yellow oil. This substance was dissolved in dichloromethane (10 ml) and TFA (5 ml) was added. This reaction mixture was stirred at room temperature for 4 days and concentrated under reduced pressure. The residue was dissolved in ethyl acetate. This mixture was washed with saturated NaHCO ₃ solution and the organic layer was dried over MgSO ₄ , filtered and evaporated to dryness under vacuum. The resulting residue was triturated with 25% ethyl acetate in hexanes and the solid was collected by filtration to give the title compound as a yellowish powder (0.506 g, yield: 17.56%). Analytical LC-MS conditions: column: Phenomenex LUNA C18, 2.0x50 mm, particle size 3 µm; mobile phase A: (10:90) methanol:water with 0.1% TFA; mobile phase B: (90:10) methanol:water with 0.1% TFA; gradient: 0100% B for 4 min, then maintain 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: 2.5 min, 267.3 (M-H ^- ). 1H NMR (400 MHz, methanol-d ₄ ) δ 7.557.43 (m, 2H), 7.36-7.27 (m, 3H), 7.27-7.20 (m, 1H), 7.08 (td, J=7.7, 1.5 Hz, 1H), 6.86-6.66 (m, 2H), 4.85 (s, 1H), 3.28 (t, J=5.3 Hz, 4H), 2.84-2.61 (m, 4H).

Intermediate 78: tert-butyl 4-((2-hydroxynaphthalene-1-yl)methyl)piperazine-1carboxylate

2-Hydroxy-1-naphthaldehyde (600 mg, 3.48 mmol) was added to a solution of tert-butyl piperazine-1-carboxylate (779 mg, 4.18 mmol) in DMF (10 ml). This reaction mixture was stirred for

- 57 044500 for 1 hour at room temperature. Sodium cyanoborohydride (657 mg, 10.45 mmol) was added and the reaction mixture was stirred at room temperature for 6 days. The reaction was stopped by adding water. The mixture was extracted with ethyl acetate (2x40 ml). The organic layers were combined, dried (MgSO ₄ ), filtered and concentrated in vacuo to give an orange oil. This substance was fractionated by reverse phase preparative HPLC using acetonitrile-water-TFA as an eluent. Homogeneous fractions were collected, neutralized with saturated NaHCO ₃ solution and then concentrated under reduced pressure to remove acetonitrile. The separated beige solid was collected by filtration and dried to obtain the title compound as a powder (498 mg, yield: 41.7%). Analytical LC-MS conditions: column: Phenomenex LUNA C18, 50x2, particle size 3 µm; mobile phase A: 5% acetonitrile: 95% water: 10 mM ammonium acetate; mobile phase B: 95% acetonitrile: 5% water: 10 mM ammonium acetate; temperature: 40°C; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: 3.6 min, 343.2 (M+H)+. 1H NMR (400 MHz, methanol-d ₄ ) δ 7.96 (d, J=8.6 Hz, 1H), 7.82-7.62 (m, 2H), 7.45 (td, J=7.7, 1.2 Hz, 1H), 7.29 (t , J=7.5 Hz, 1H), 7.06 (d, J=9.0 Hz, 1H), 4.17 (s, 2H), 3.60-3.43 (m, 4H), 2.63 (t, J=5.0 Hz, 4H), 1.48 (s, 9H).

Intermediate 79: 1-(piperazin-1-ylmethyl)naphthalene-2-ol bis(2,2,2-trifluoroacetate)

To a solution of tert-butyl 4-((2-hydroxynaphthalene-1-yl)methyl)piperazine-1-carboxylate (494 mg, 1.443 mmol) in dichloromethane (3 mL) was added TFA (2 mL, 26.0 mmol). This reaction mixture was stirred overnight at room temperature and then concentrated under reduced pressure to obtain the title compound as a purple solid (670 mg, yield: 99%). Analytical LC-MS conditions: column: Phenomenex LUNA C18, 2.0x50 mm, particle size 3 µm; mobile phase A: (10:90) methanol:water with 0.1% TFA; mobile phase B: (90:10) methanol:water with 0.1% TFA; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: 2.1 min, 243.0 (M+H)+. 1H NMR (400 MHz, methanol^) δ 8.03 (d, J=8.6 Hz, 1H), 7.91 (d, J=8.8 Hz, 1H), 7.85 (d, J=8.1 Hz, 1H), 7.64-7.54 ( m, 1H), 7.39 (t, J=7.5 Hz, 1H), 7.23 (d, J=8.8 Hz, 1H), 4.84 (s, 2H), 3.67-3.56 (m, 4H), 3.55-3.45 (m , 4H).

Intermediate 80: 1-(2-(4-fluorophenyl)propan-2-yl)-4-tosylpiperazine

A mixture of 2-(4-fluorophenyl)propan-2-amine hydrochloride (0.5 g, 2.64 mmol), N,N-bis(2-chloroethyl)-4methylbenzenesulfonamide (0.820 g, 2.77 mmol) and DIPEA ( 1.381 ml, 7.91 mmol) was heated using microwave radiation for 20 hours at 125°C. Water was added and the mixture was extracted using dichloromethane (2x30 ml). The organic layers were combined, dried (MgSO ₄ ), filtered and concentrated under reduced pressure to give a yellow oil. This oil was triturated with 25% ethyl acetate in hexanes and the resulting yellowish solid was collected by filtration to give the title compound (505 mg, yield: 50.9%). Analytical LC-MS conditions: Phenomenex LUNA C18, 50x2, particle size 3 µm; mobile phase A: 5% acetonitrile: 95% water: 10 mM ammonium acetate; mobile phase B: 95% acetonitrile: 5% water: 10 mM ammonium acetate; temperature: 40°C; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: 3.7 min, 377.1 (M+H) ⁺ . 1H NMR (400 MHz, DMSO-d ₆ ) δ 7.61 (d, J=8.1 Hz, 2H), 7.47 (d, J=8.1 Hz, 2H), 7.40 (dd, J=8.6, 5.9 Hz, 2H), 7.07 (t, J=8.8 Hz, 2H), 2.83 (br. s, 4H), 2.44 (s, 7H), 1.25 (s, 6H).

Intermediate 81: 1-(2-(4-fluorophenyl)propan-2-yl)piperazine

A solution of hydrogen bromide (6 ml, 33.1 mmol, 33% by weight) in acetic acid was added to a mixture of 1-(2-(4-fluorophenyl)propan-2-yl)-4-tosylpiperazine (495 mg, 1.315 mmol ) and 4-hydroxybenzoic acid (545 mg, 3.94 mmol). This reaction mixture was stirred at room temperature over the weekend. Water was then slowly added and the resulting mixture was stirred for 2 hours. The resulting white precipitate was collected by filtration. The precipitate was washed successively with cold water and then with toluene. The aqueous filtrate and washing water were then cooled in

- 58 044500 ice bath and made alkaline to pH > 10 by adding NaOH granules. This mixture was extracted with ethyl acetate (2x20 ml) and the combined extracts were washed with brine, dried (MgSO ₄ ), filtered and concentrated under reduced pressure to obtain the title compound as a yellowish solid (195 mg, yield: 66.7%). Analytical LC-MS conditions: Phenomenex LUNA C18, 50x2, particle size 3 µm; mobile phase A: 5% acetonitrile: 95% water: 10 mM ammonium acetate; mobile phase B: 95% acetonitrile: 5% water: 10 mM ammonium acetate; temperature: 40°C; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: 1.8 min, 223.1 (M+H) ⁺ . 1H NMR (400 MHz, chloroform-d) δ 7.51 (dd, J=8.6, 5.6 Hz, 2H), 6.99 (t, J=8.7 Hz, 2H), 2.96-2.73 (m, 4H), 2.52-2.36 ( m, 4H), 1.34 (s, 6H).

Intermediate 82: 1-(1-(4-fluorophenyl)cyclopropyl)-4-tosylpiperazine

Intermediate 82 was prepared following the general procedure used for the synthesis of 1-(2-(4-fluorophenyl)propan-2-yl)piperazine, except that 1-(4fluorophenyl)cyclopropan-1-amine hydrochloride (0 .5 g, 2.66 mmol). The title compound was synthesized as a light yellow solid (651 mg, yield: 65.2%). Analytical LC-MS conditions: Phenomenex LUNA C18, 50x2, particle size 3 µm; mobile phase A: 5% acetonitrile: 95% water: 10 mM ammonium acetate; mobile phase B: 95% acetonitrile: 5% water: 10 mM ammonium acetate; temperature: 40°C; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: 3.6 min, 375.1 (M+H)+. ^1H NMR (400 MHz, chloroform-d) δ 7.61 (d, J=8.3 Hz, 2H), 7.31 (d, J=8.3 Hz, 2H), 7.20 (dd, J=8.4, 5.5 Hz, 2H), 7.01 (t, J=8.6 Hz, 2H), 3.15-2.78 (m, 4H), 2.58 (b. m, 4H), 2.44 (s, 3H), 0.90-0.72 (m, 4H).

Intermediate 83: 1-(1-(4-fluorophenyl)cyclopropyl)piperazine

Intermediate 83 was prepared following the general procedure used for the synthesis of 1-(2-(4-fluorophenyl)propan-2-yl)piperazine, except that 1-(1-(4fluorophenyl)cyclopropyl)-4- tosylpiperazine (600 mg, 1.602 mmol), this compound was synthesized as a brown solid (235 mg, yield: 66.6%). Analytical LC-MS conditions: Phenomenex LUNA C18, 50x2, particle size 3 µm; mobile phase A: 5% acetonitrile: 95% water: 10 mM ammonium acetate; mobile phase B: 95% acetonitrile: 5% water: 10 mM ammonium acetate; temperature: 40°C; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: 1.8 min, 221.1 (M+H) ⁺ . 1H NMR (400 MHz, methanol-d ₄ ) δ 7.34 (br. s, 2H), 7.06 (br. d, J=2.0 Hz, 2H), 3.01-2.13 (m, 8H), 1.20-0.37 (m, 4H).

Intermediate 84: 1-(1-(4-fluorophenyl)-2-methylpropyl)piperazine bis(2,2,2-trifluoroacetate)

To a solution of 1-(4-fluorophenyl)-2-methylpropan-1-one (250 mg, 1.504 mmol) in THF (4 ml) was added titanium (IV) chloride (1.0 N solution in dichloromethane) (1.956 ml, 1.956 mmol). This reaction mixture was stirred for 0.5 h at room temperature. tert-Butyl piperazine 1-carboxylate (336 mg, 1.805 mmol) in THF (4 ml) was added. The color of the reaction mixture changed from green to yellow. The reaction mixture was stirred for 1 hour and then sodium cyanoborohydride (123 mg, 1.956 mmol) was added and stirring was continued for an additional 3 days. The reaction was stopped by adding acetic acid. The mixture was diluted by adding ethyl acetate. The resulting solution was washed with brine and the organic layer was separated, dried (MgSO ₄ ), filtered and evaporated under reduced pressure to obtain a viscous yellow oil. This crude product was dissolved in dichloromethane (4 ml) and TFA (2 ml) was added. The reaction mixture was then stirred overnight at room temperature and then concentrated to dryness under reduced pressure. The residue was fractionated by reverse phase preparative HPLC using acetonitrile-water-TFA as the eluent. Homogeneous fractions were combined and concentrated in vacuo to give the title compound as a white solid (103 mg, yield: 14.74%). Analytical LC-MS conditions: Phenomenex LUNA C18, 50x2, particle size 3 µm; mobile phase A: 5% acetonitrile: 95% water: 10 mM ammonium acetate; mobile phase B: 95% aceto- 59 044500 nitrile: 5% water: 10 mM ammonium acetate; temperature: 40°C; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. results

LC-MS: 2.3 min, 237.1 (M+H)+. ¹ H NMR (400 MHz, methanol^) δ 7.28 (dd, J=8.6, 5.4 Hz, 2H), 7.19-7.01 (m,

2H), 3.33-3.17 (m, 5H), 2.78-2.60 (br. m, 4H), 2.36 (dt, J=9.5, 6.6 Hz, 1H), 1.07 (d, J=6.6 Hz, 3H), 0.76 (d,

J=6.6 Hz, 3H).

Intermediate 85: 1-(cyclobutyl(4-fluorophenyl)methyl)piperazine bis(2,2,2-trifluoroacetate)

Intermediate 85 was prepared following the general procedure used for the synthesis of 1-(2-(4-fluorophenyl)propan-2-yl)piperazine, except that cyclobutyl(4-fluorophenyl)methanone (300 mg, 1.683 mmol) was used ). The title compound was synthesized as a white solid (40.2 mg, yield: 5.01%). Analytical LC-MS conditions: Phenomenex LUNA C18, 50x2, particle size 3 µm; mobile phase A: 5% acetonitrile: 95% water: 10 mM ammonium acetate; mobile phase B: 95% acetonitrile: 5% water: 10 mM ammonium acetate; temperature: 40°C; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm.

LC-MS results: 2.3 min, 249.1 (M+H) ⁺ . 1H NMR (400 MHz, methanol-d4) δ 7.48 (dd, J=8.6, 5.4 Hz, 2H), 7.20 (t, J=8.6 Hz, 2H), 4.22 (d, J=10.5 Hz, 1H), 3.54 -3.43 (m, 4H), 3.41-3.28 (m, 2H), 3.27-3.11 (m, 3H), 2.55-2.29 (m, 1H), 2.16 (quin, J=9.8 Hz, 1H), 2.06-1.88 (m, 1H), 1.86-1.72 (m, 1H), 1.71-1.61 (m, 1H), 1.601.46 (m, 1H).

Intermediate 86: tert-butyl 4-(3-cyano-1,6-dimethyl-2-oxo-1,2-dihydro-1,5naphthyridin-4-yl)piperazine-1-carboxylate

Intermediate 86 was prepared following the general procedures described for Intermediates 55 and 61. LC-MS. [Injection volume = 3 µl, initial %B = 2, final %B =98, gradient duration = 1.5 min, flow rate = 0.8 ml/min, wavelength = 220 nm, solvent pair = water/acetonitrile /TFA, solvent A = 100% water/0.05% TFA, solvent B = 100% acetonitrile/0.05% TFA, column = Waters Aquity VEN C18 2.1x50 mm, 1.7 µm MW1, oven temperature = 40] RT = 1.170 min. (M+H)+ = 384.05.

Intermediate 87: 1,6-dimethyl-2-oxo-4-(piperazin-1-yl)-1,2-dihydro-1,5-naphthyridine-3-carbonitrile 2,2,2-trifluoroacetate

CH ₃

XXX

H ₃ C CN o N CF ₃ CO ₂ H Η (1-87) tert-Butyl 4-(3-cyano-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridine- 4-yl)piperazine-1-carboxylate (26 mg, 0.068 mmol) was dissolved in DCM (1 ml) and trifluoroacetic acid (1.045 ml, 13.56 mmol) was added. This mixture was stirred at room temperature for approximately 1 hour. The solvent and excess acid were removed in vacuo to give the desired product as an oil. LC-MS. [Injection volume = 3 µl, initial % B = 2, final % B = 98, gradient duration = 1.5 min, flow rate = 0.8 ml/min, wavelength = 220 nm, solvent pair = water/acetonitrile /TFA, solvent A = 100% water/0.05% TFA, solvent B = 100% acetonitrile/0.05% TFA, column = Waters Aquity VEN C18 2.1x50 mm, 1.7 µm MW1, oven temperature = 40 ] RT = 0.799 min. (M+H)+ = 284.05 (free base).

Intermediate 88: tert-butyl (1R,5S)-3-(6-chloro-3-cyano-1-methyl-2-oxo-1,2-dihydro- 60 044500

1,5-naphthyridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate

СН ₃ dХХ CK N y CN ,ν сн ₃ ο^ _ο 4όη ₃ ^СН з (1-88)

To a solution of 4,6-dichloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (164 mg, 0.645 mmol) and triethylamine (0.270 ml, 1.936 mmol) in DMF ( 2 ml) (1R,5S)-tert-butyl 3,8diazabicyclo[3.2.1]octane-8-carboxylate (137 mg, 0.645 mmol) was added and the resulting mixture was stirred overnight at room temperature. The crude reaction mixture was adsorbed onto silica gel and subjected to flash chromatography using EtOAc as eluent. Homogeneous fractions were combined and evaporated in vacuo to obtain the desired product as a yellow solid (200 mg). LC-MS. [Injection volume = 3 µl, initial %B = 2, final %B =98, gradient duration = 1.5 min, flow rate = 0.8 ml/min, wavelength = 220 nm, solvent pair = water/acetonitrile /TFA, solvent A = 100% water/0.05% TFA, solvent B = 100% acetonitrile/0.05% TFA, column = Waters Aquity VEN C18 2.1x50 mm, 1.7 µm MW1, oven temperature = 40 ] RT = 1.260 min. (M-tert-Bu+H) ⁺ = 373.95. 1H NMR (500 MHz, DMSOZH) δ 8.11 (d, J=9.1 Hz, 1H), 7.84 (d, J=9.0 Hz, 1H), 4.25 (br. s., 2H), 4.12 (s, 2H), 3.61-3.47 (m, 4H), 2.22 (d, J=7.6 Hz, 2H), 1.88-1.80 (m, 2H), 1.46 (s, 9H).

Intermediate 89: 4-((1R,5S)-3,8-diazabicyclo[3.2.1]octan-3-yl)-6-chloro-1-methyl-2oxo-1,2-dihydro-1,5- naphthyridine-3-carbonitrile 2,2,2-trifluoroacetate

CH ₃ dXX CK N y CN 6th cf ₃ co ₂ h _(M9)

Trifluoroacetic acid (0.717 ml, 9.30 mmol) was diluted with DCM (0.8 ml) and the resulting mixture was added to the flask containing (1R,5S)-tert-butyl 3-(6-chloro-3-cyano-1 -methyl-2-oxo1,2-dihydro-1,5-naphthyridin-4-yl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (200 mg, 0.465 mmol). The resulting yellow solution was stirred for 30 minutes at room temperature and then evaporated in vacuo to give the title compound as a brown oil. LC-MS. [Injection volume = 3 µl, initial %B = 2, final %B =98, gradient duration = 1.5 min, flow rate = 0.8 ml/min, wavelength = 220 nm, solvent pair = water/acetonitrile /TFA, solvent A = 100% water/0.05% TFA, solvent B = 100% acetonitrile/0.05% TFA, column = Waters Aquity VEN C18 2.1x50 mm, 1.7 µm MW1, oven temperature = 40 ] RT = 0.756 min, (M + N) ⁺ = 331.00. ^1H NMR (DMSOZH) δ 8.14(d, J=9.0 Hz, 1H), 7.87 (d, J=8.8 Hz, 1H), 4.23 (br. d, J=11.2 Hz, 4H), 3.84 (br. d , J=13.0 Hz, 2H), 3.57 (s, 3H), 2.42 (br. d, J=7.8 Hz, 2H), 1.84-2.03 (m, 2H).

Intermediate 90: Cyclohexyl(phenyl)methanol

HE

ΓΥιΊ (1-90)

Sodium borohydride (0.201 g, 5.31 mmol) was added to a solution of cyclohexyl (phenyl)methanone (1 g, 5.31 mmol) in ethanol (20 ml) and the mixture was stirred overnight at room temperature. Then the reaction mixture was evaporated under vacuum. The residue was dissolved in DCM and washed successively with water and brine. The organic layer was then dried over MgSO ₄ , filtered and evaporated under reduced pressure to obtain the desired product as a colorless oil (1.01 g). LC-MS. [Injection volume = 3 µl, initial %B = 2, final %B =98, gradient duration = 1.5 min, flow rate = 0.8 ml/min, wavelength = 220 nm, solvent pair = water/acetonitrile /TFA, solvent A = 100% water/0.05% TFA, solvent B = 100% acetonitrile/0.05% TFA, column = Waters Aquity VEN C18 2.1x50 mm, 1.7 µm MW1, oven temperature = 40 ] RT = 1.255 min. There were no components with MWt < 200 in the mass spectrum. 1H NMR (400 MHz, DMSOd6) δ 7.34-7.17 (m, 5H), 5.01 (d, J=4.4 Hz, 1H), 4.23 (dd, J=6.4, 4.6 Hz, 1H), 1.89-0.85 (m, 11H).

Intermediate 91: Bromo(cyclohexyl)methyl)benzene

- 61 044500

To a mixture of pentabromophosphorane (3.43 g, 7.96 mmol) in DCM (20 mL), a solution of cyclohexyl(phenyl)methanol (1.01 g, 5.31 mmol) in DCM (10 mL) was slowly added at room temperature. After 1 hour, the reaction mixture was washed with water (1 x 30 ml), 10% NaHSO ₃ solution (1 x 25 ml) and water (1 x 30 ml). The resulting material was dried over MgSO ₄ , filtered and evaporated under reduced pressure to obtain the desired product as a colorless oil (1.32 g). LC-MS. [Injection volume = 3 µl, initial %B = 2, final %B =98, gradient duration = 1.5 min, flow rate = 0.8 ml/min, wavelength = 220 nm, solvent pair = water/acetonitrile /TFA, solvent A = 100% water/0.05% TFA, solvent B = 100% acetonitrile/0.05% TFA, column = Waters Aquity VEN C18 2.1x50 mm, 1.7 µm MW1, oven temperature = 40 ] RT = 1.609 min. The ionization conditions used did not allow mass determination. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 7.52-7.18 (m, 5H), 5.03 (d, J=9.3 Hz, 1H), 2.26-0.75 (m, 11H).

Intermediate 92: tert-butyl 4-(cyclohexyl(phenyl)methyl)piperazine-1-carboxylate O_O,/ ^CH3

I GSNz N CH ₃ (1-92)

A mixture of potassium carbonate (218 mg, 1.580 mmol), tert-butyl piperazine-1-carboxylate (147 mg, 0.790 mmol) and (bromo(cyclohexyl)methyl)benzene (200 mg, 0.790 mmol) in acetonitrile (10 ml) was heated at reflux temperature in a nitrogen atmosphere overnight. NaI (~2 mg) was added and the mixture was heated at reflux for an additional 24 hours. The reaction mixture was then filtered and the filtrate evaporated to dryness. The residue was dissolved in 30% EtOAc in hexanes and the mixture was filtered through a pad of silica gel. The eluted material was evaporated under reduced pressure and the residue was placed under high vacuum for 1 hour to give the title compound (223 mg). LC-MS. [Injection volume = 3 µl, initial % B = 2, final % B = 98, gradient duration = 1.5 min, flow rate = 0.8 ml/min, wavelength = 220 nm, solvent pair = water/acetonitrile /TFA, solvent A = 100% water/0.05% TFA, solvent B = 100% acetonitrile/0.05% TFA, column = Waters Aquity VEN C18 2.1x50 mm, 1.7 µm MW1, oven temperature = 40 ] RT = 0.857 min. (M-Boc+AcOH+ACN)+ = 403.25.

Intermediate 93: 1-(cyclohexyl(phenyl)methyl)piperazine bis(2,2,2-trifluoroacetate)

N (CP ₃ CO ₂ H) ₂

NN (1-93)

Trifluoroacetic acid (0.479 ml, 6.22 mmol) was added to a solution of tert-butyl 4(cyclohexyl(phenyl)methyl)piperazine-1-carboxylate (223 mg, 0.622 mmol) in DCM (0.5 ml) and the resulting mixture was stirred in for 1 hour at room temperature. Then the reaction mixture was evaporated to dryness. LC-MS. [Injection volume = 3 µl, initial %B = 2, final %B =98, gradient duration = 1.5 min, flow rate = 0.8 ml/min, wavelength = 220 nm, solvent pair = water/acetonitrile /TFA, solvent A = 100% water/0.05% TFA, solvent B = 100% acetonitrile/0.05% TFA, column = Waters Aquity VEN C18 2.1x50 mm, 1.7 µm MW1, oven temperature = 40 ] RT = 0.850 min. (M+H)+ = 259.1 (free base).

Intermediate 94: tert-butyl 4-(2-hydroxybenzyl)piperazine-1-carboxylate

Acetic acid (0.307 mL, 5.37 mmol) was added to a solution of tert-butyl piperazine-1-carboxylate (1 g, 5.37 mmol) and 2-hydroxybenzaldehyde (0.656 g, 5.37 mmol) in DCE through pore size molecular sieves 4 microns. This mixture was stirred for 1 hour at room temperature and sodium triacetoxyborohydride (0.993 mL, 5.37 mmol) was added. The reaction mixture was stirred over the weekend, then the reaction was stopped by adding MeOH (10 ml). The reaction mixture was stirred for 30 minutes and filtered. The resulting filtrate was concentrated under vacuum.

- 62 044500

The crude residue was purified by column chromatography using 5-50% EtOAc in hexanes as eluent. Homogeneous fractions were combined and concentrated under reduced pressure to give the title compound as a pale yellow oil (684 mg). LC-MS. [Injection volume = 3 µl, initial % B = 2, final % B = 98, gradient duration = 1.5 min, flow rate = 0.8 ml/min, wavelength = 220 nm, solvent pair = water/acetonitrile /TFA, solvent A = 100% water/0.05% TFA, solvent B = 100% acetonitrile/0.05% TFA, column = Waters Aquity VEN C18 2.1x50 mm, 1.7 µm MW1, oven temperature = 40 ] RT = 0.810 min. (M+H)+ = 293.40. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.11-9.89 (s, 1H), 7.20-7.01 (m, 2H), 6.85-6.69 (m, 2H), 3.60 (s, 2H), 3.41-3.27 (m, 4H), 2.39 (t, J=4.9 Hz, 4H), 1.48-1.34 (m, 9H).

Intermediate 95: 2-(piperazin-1-ylmethyl)phenol bis(2,2,2-trifluoroacetate) (CF ₃ CO ₂ H) ₂ OH ^ΗΝ ^ (1-95)

Trifluoroacetic acid (1.792 ml, 23.26 mmol) was added to a solution of tert-butyl 4-(2-hydroxybenzyl)piperazine-1-carboxylate (680 mg, 2.326 mmol) in DCM (2 ml) and the resulting mixture was stirred for 1 h at room temperature. The mixture was then concentrated in vacuo to give the desired product as a red oil which slowly crystallized over time. LC-MS. [Injection volume = 3 µl, initial % B = 2, final % B = 98, gradient duration = 1.5 min, flow rate = 0.8 ml/min, wavelength = 220 nm, solvent pair = water/acetonitrile /TFA, solvent A = 100% water/0.05% TFA, solvent B = 100% acetonitrile/0.05% TFA, column = Waters Aquity VEN C18 2.1x50 mm, 1.7 µm MW1, oven temperature = 40 ] RT = 0.599 min.

Intermediate 96: 8-hydroxy-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile

CH ₃ chi NC Ν

HE (1-96)

8-Hydroxy-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (1 g, 4.97 mmol) was suspended in acetonitrile (25 ml) in a round bottom flask equipped with a reflux condenser. POCl ₃ (6.02 mL, 64.6 mmol) was added to this solution. The reaction mixture was heated under nitrogen atmosphere for 3.5 hours at 85°C. Ice was added to the reaction mixture and then 5N. an aqueous solution of sodium hydroxide and a saturated solution of sodium bicarbonate until the evolution of CO ₂ gas ceased. The aqueous fraction was extracted with chloroform. The organic fractions were combined and washed successively with 1.5 M K ₂ HPO ₄ solution and saline. The organic fraction was dried over sodium sulfate, then the drying agent was filtered from the extract and the solvent was removed under vacuum using a rotary evaporator. The title compound (958 mg, yield: 72%) was isolated as a yellow-green solid. LC-MS analysis confirmed the desired product. Gradient duration = 4 min, flow rate = 0.8 ml/min, wavelength = 220, solvent pair = ACN: water: ammonium acetate, solvent A = 5% ACN: 95% water: 10 mM ammonium acetate, solvent B = 95% ACN: 5% water: 10 mM ammonium acetate, column = Phenomenex LUNA C18, 50x2.3 µm, oven temperature = 40. LC-MS results: 1.9 min, 220.0 (M+H) ⁺ .

Intermediate 97: tert-butyl 4-(6-bromo-3-cyano-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridin-4-yl)piperazine-1-carboxylate

CH ₃

XXX BrN J CN O CH ₃ N H3C^O^O CH ₃ (1-97)

Tert. -butyl piperazine-1-carboxylate (3.12 g, 16.75 mmol) and Huenig's base (5.85 ml, 33.5 mmol). A large amount of substance immediately precipitated. According to LC-MS analysis, the reaction was complete. The resulting precipitate was collected and washed with diethyl ether, EtOAC and DCM to give the title compound (6.2 g, yield: 83%). LC-MS analysis was used to identify and determine the purity of the resulting product. Conditions for analysis of the introduced sample: column: Waters Aquity VEN C18, 2.1 mmx50 mm, particle size 1.7

- 63 044500 microns; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B:

(95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 40°C; gradient: from 0% B to

100% B for 1.5 min, then maintain 100% B for 0.75 min; flow rate: 1 ml/min;

registration: MS and UV (220 nm). Results of the analysis of the injected sample: purity: >90%; measured mass: 448.1; retention time: 1.4 min.

Intermediate 98: tert-butyl 4-(3-cyano-6-methoxy-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridin-4-yl)piperazine-1-carboxylate

Hermetically sealed reaction tube containing tert-butyl 4-(6-bromo-3-cyano-1methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)piperazin-1-carboxylate (500 mg , 1.115 mmol), palladium (II) acetate (12.52 mg, 0.056 mmol), cesium carbonate (363 mg, 1.115 mmol) and 5-[di(1-adamantyl)phosphino]-1',3',5' -triphenyl-1'h-[1,4']bipyrazole (73.9 mg, 0.112 mmol) was placed under vacuum and then sealed under nitrogen. Methanol (0.2 ml) and acetonitrile (4 ml) were added and the reaction mixture was heated at 80°C overnight. According to LC-MS analysis, the reaction was complete. The mixture was diluted with ethyl acetate, filtered, concentrated and the residue was purified using silica gel chromatography (hexane/ethyl acetate, silica gel column (40 g), 30-100% EtOAc). The title compound was isolated as a yellow solid (360 mg, yield: 81%). Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 400.05; retention time: 1.69 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 im). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 400.06; retention time: 1.72 min.

Intermediate 99: 6-methoxy-1-methyl-2-oxo-4-(piperazin-1-yl)-1,2-dihydro-1,5naphthyridine-3-carbonitrile

To a solution of tert-butyl 4-(3-cyano-6-methoxy-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)piperazine-1-carboxylate (350 mg, 0.876 mmol) in dichloromethane (3 ml) was added trifluoroacetic acid (0.675 ml, 8.76 mmol). This reaction mixture was stirred for 2 hours at room temperature. According to LC-MS analysis, the reaction was complete. The solvent was removed and the crude residue was diluted with ethyl acetate. This solution was washed with sodium bicarbonate and brine and dried over sodium sulfate to obtain a yellow solid (250 mg, yield: 90%). Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 300.11; retention time: 0.95 min. Analysis conditions for the 2nd sample injection: column: Waters Bridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration 64 044500: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 94.8%; measured mass: 300.09; retention time: 0.81 min.

Intermediate 100: (4-fluorophenyl)(2-methoxypyridin-3-yl)methanol

2-Methoxynicotinaldehyde (2.5 g, 18.23 mmol) was dissolved in THF (36.5 mL) in a 500 mL round bottom flask and cooled in an ice bath. After 5 minutes, (4fluorophenyl)magnesium bromide in diethyl ether (10.94 mL, 21.88 mmol) was added by syringe and the reaction mixture was stirred for 50 minutes. The cooling bath was removed and the reaction mixture was stirred for 10 minutes. The reaction was stopped by sequentially adding 1 ml of saturated aqueous ammonium chloride and 50 ml of ethyl acetate. The solid was removed by decantation. The solvent was removed by rotary evaporation and the crude residue was chromatographed on silica gel using 5-15% methanol in ethyl acetate. Fractions containing the desired product were combined. Removal of the solvent gave (4-fluorophenyl)(2-methoxypyridin-3-yl)methanol (3 g, 12.22 mmol, yield: 67.0%) as a yellow solid. According to NMR analysis, the purity of this substance was > 95%. ¹ H NMR (400 MHz, chloroform-d) δ 8.12 (d, J=4.9 Hz, 1H), 7.54 (d, J=7.1 Hz, 1H), 7.37 (dd, J=8.3, 5.6 Hz, 2H), 7.05 (t, J=8.6 Hz, 2H), 6.91 (dd, J=7.3, 5.1 Hz, 1H), 5.98 (d, J=4.6 Hz, 1H), 3.98 (s, 3H), 2.92 (d, J =4.6 Hz, 1H).

Intermediate 101: (cyanomethyl)trimethylphosphonium iodide

H ₃ C

1' ₊ CH ₃ NC^P^ ³ 1' ^sn s (1-101)

The title compound was prepared according to the method of Zaragoza and Stephensen (J. Org. Chem. 2001, 66, 2518-2521). Trimethyl phosphane in toluene (80 mL, 80 mmol) was diluted by adding THF (40 mL) and toluene (40 mL) in a 1 L round bottom flask and cooled in an ice bath. (Cyanomethyl)trimethylphosphonium iodide was added dropwise to the reaction mixture with constant vigorous stirring to obtain a yellowish-brown precipitate. The cooling bath was removed and the reaction mixture was stirred overnight at room temperature. The reaction mixture was placed in an ultrasonicator to break up lumps of the solid, and then the reaction mixture was stirred for an additional 4 hours. The solid was collected by filtration and dried under vacuum to give the title compound (17.1 g, 88%). NMR analysis confirmed the receipt of pure desired product. 1H NMR (400 MHz, DMSO-d ₆ ) δ 4.03 (d, J=16.4 Hz, 2H), 2.05 (d, J=15.4 Hz, 9H).

Intermediate 102: (4-fluorophenyl)(3-methoxypyridin-2-yl)methanol

Intermediate 102 was prepared from suitable starting materials according to the general procedure described for Intermediate 100. 1H NMR (500 MHz, chloroform-d) δ 8.21 (dd, J=4.8, 1.1 Hz, 1H), 7.39-7.32 (m , 2H), 7.25 (dd, J=8.3, 4.8 Hz, 1H), 7.15 (dd, J=8.2, 1.1 Hz, 1H), 7.016.94 (m, 2H), 5.94 (d, J=6.9 Hz, 1H), 5.50 (d, J=7.0 Hz, 1H), 3.79 (s, 3H).

Intermediate 103: (4-fluorophenyl)(pyridin-2-yl)methanol

Intermediate 103 was prepared from appropriate starting materials according to the general procedure described for Intermediate 100. 1H NMR (500 MHz, chloroform-d) δ 8.59 (d, J=4.9 Hz, 1H), 7.65 (td, J=7.6 , 1.7 Hz, 1H), 7.36 (dd, J=8.6, 5.4 Hz, 2H), 7.23 (dd, J=7.2, 5.0 Hz, 1H), 7.14 (d, J=7.8 Hz, 1H), 7.04 (t , J=8.7 Hz, 2H), 5.75 (d, J=2.0 Hz, 1H), 5.43-5.14 (m, 1H).

Intermediate 104: (4-fluoro-2-methoxyphenyl)(pyrimidin-2-yl)methanol

F (1-104)

Intermediate 104 was prepared from suitable starting materials according to the general procedure described for intermediate 100. 1H NMR (500 MHz, chloroform-d) δ 8.77 (d, J=4.9 Hz, 2H), 7.24 (t , J=4.9 Hz, 1H), 7.18 (dd, J=8.9, 6.7 Hz, 1H), 6.69-6.61 (m, 2H), 6.15 (d, J=5.4 Hz, 1H),

4.77 (d, J=5.6 Hz, 1H), 3.79 (s, 3H).

Intermediate 105: bis(4-fluoro-2-methoxyphenyl)methanol

H ₃ C. _ , ,sn ₃ ³ O he o ³

Intermediate 105 was prepared from suitable starting materials according to the general procedure described for intermediate 100. Ή NMR (500 MHz, chloroform-d) δ 7.17 (dd, J=8.9, 7.0 Hz, 2H), 6.72-6.58 (m , 4H), 6.26 (d, J=3.2 Hz, 1H), 3.83 (s, 6H), 3.26 (br. d, J=4.4 Hz, 1H).

Intermediate 106: tert-butyl 4-(3,6-dicyano-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridin-4-yl)piperazine-1-carboxylate

CH ₃

In a hermetically sealed reaction tube, tert-butyl 4-(6-bromo-3-cyano-1methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)piperazin-1-carboxylate (150 mg) was mixed , 0.335 mmol), zinc (2.92 mg, 0.045 mmol), zinc cyanide (15.72 mg, 0.134 mmol) and complex of 1,1'-bis(diphenylphosphino)ferrocene-palladium (II) dichloride and dichloromethane (54, 6 mg, 0.067 mmol). This reaction mixture was placed under vacuum and then sealed under a nitrogen atmosphere. The solid was suspended in NMP (2 ml) from a new, previously unopened vial. The reaction tube was heated for 2.5 hours at 80°C. According to LC-MS analysis, the reaction was complete. The crude material was purified by preparative HPLC (MeOH/water) using TFA as a buffer to give the desired product as a yellow solid (95 mg, 67%). Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for 1st sample input: column:

Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 microns; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 98.1%; measured mass: 395.12; retention time: 1.66 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 99.4%; measured mass: 395.15; retention time: 1.66 min. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ 8.26 (d, J=8.9 Hz, 1H), 8.16 (d, J=8.8 Hz, 1H), 3.83 (br. s, 4H), 3.69-3.36 ( m, 3H), 1.44 (s, 9H).

Intermediate 107: 5-methyl-6-oxo-8-(piperazin-1-yl)-5,6-dihydro-1,5-naphthyridine2,7-dicarbonitrile

Intermediate 107 was prepared from tert-butyl 4-(3,6-dicyano-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridin-4-yl)piperazin-1-carboxylate according to the general procedure, described for Intermediate 99. Analytical LC-MS was used to determine the final purity of the product. Conditions for analysis of the injected sample: column: Waters Aquity VEN C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 40°C; gradient: from 0% B to 100% B for 1.5 min, then maintaining 100% B for 0.75 min; flow rate 66 044500: 0.8 ml/min; registration: MS and UV (220 nm). Results of the analysis of the introduced sample: purity: 91%;

measured mass: 294.8; retention time: 0.87 min.

Intermediate 108: (4-fluoro-2-methoxyphenyl)(4-fluorophenyl)methanol

OH O'™’ w,™

Intermediate 108 was prepared from suitable starting materials according to the general procedure described for Intermediate 100. 1H NMR (400 MHz, chloroform-d) δ 7.35 (dd, J=8.3, 5.6 Hz, 2H), 7.20 (t, J =7.6 Hz, 1H), 7.03 (t, J=8.7 Hz, 2H), 6.71-6.58 (m, 2H), 6.03 (d, J=4.6 Hz, 1H), 3.83 (s, 3H), 2.78 (d , J=4.9 Hz, 1H, OH).

Intermediate 109: Ethyl 4-((4-fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl)piperazine 1-carboxylate

ABOUT

F 11

N O CH ₃ y ^CH3

F (1-109)

Intermediate 109 was prepared from suitable starting materials according to the general procedure described for intermediate 100. 1H NMR (400 MHz, chloroform-d) δ 7.58-7.46 (m, 1H), 7.36 (dd, J=8.3, 5.6 Hz , 2H), 6.96 (t, J=8.6 Hz, 2H), 6.67 (td, J=8.3, 2.2 Hz, 1H), 6.56 (dd, J=10.8, 2.2 Hz, 1H), 4.74 (s, 1H) , 4.19-4.08 (m, 2H), 3.79 (s, 3H), 3.47 (b.t, J=4.6 Hz, 4H), 2.46-2.24 (m, 4H), 1.26 (t, 7= 7.1 Hz, 3H ).

Intermediates 110 and 111 1-((4-fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl)piperazine n

V o' ^СНз сЛЛ (1-1У and 1-111)

Ethyl 4-((4-fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl)piperazine-1-carboxylate (3.6 g, 9.22 mmol) was mixed with water (23.05 ml) and methanol (69, 2 ml) in a 500 ml round bottom flask. Potassium hydroxide (7.76 g, 138 mmol) was then added. This reaction mixture was heated at reflux while monitoring the progress of the reaction using LC/MS analysis. After heating for 7 days at reflux temperature, the reaction seemed complete. The reaction mixture was extracted with diethyl ether (5 x 30 ml). The combined organic extracts were concentrated, redissolved in diethyl ether and dried over magnesium sulfate. Removal of the solvent gave 2.9 g of a pale yellow oil, which was dried under high vacuum overnight. LC/MS and NMR analysis confirmed the receipt of the desired racemic product. Four extra-hydrogens were present in the aliphatic region of the NMR spectrum. 1H NMR (500 MHz, chloroform-d) δ 7.54 (t, J=7.8 Hz, 1H), 7.36 (dd, J=8.2, 5.6 Hz, 2H), 6.95 (t, J=8.5 Hz, 2H), 6.66 (td, J=8.3, 2.3 Hz, 1H), 6.55 (dd, J=10.9, 2.4 Hz, 1H), 4.71 (s, 1H), 3.79 (d, J=1.1 Hz, 3H), 2.90 (t, J=4.8 Hz, 4H), 2.50-2.25 (m, 4H), 1.68 (br. s, 3H). Enantiomers were separated by chiral chromatography using the following conditions: column: Chiralpak AD-H, 30x250 mm, 5 µm, mobile phase: 10% MeOH/0.2% DEA/90% CO ₂ , pressure: 150 bar (1.5- 107 Pa), temperature: 30°C, flow rate: 100 ml/min, UV: 275 nm, injected sample volume: 0.5 ml (~75 mg/ml in a mixture of EtOH:CHCl ₃ (~9:1)) , fraction collection: Slope and Level mode without auxiliary gas consumption, first eluted enantiomer: 4.00'-6.00', second eluted enantiomer: 5.30' - 10.00'.

Intermediate 110 (1.1 g, yield: 75%) was isolated as the first enantiomer to elute. Intermediate 111 (1.1 g, yield: 75%) was isolated as the second eluting enantiomer.

Example 1 Ethyl 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-6-bromo-1-methyl-2-oxo-1,2-dihydro1,5-naphthyridine-3-carboxylate

- 67 044500

To a solution of ethyl 6-bromo-4-hydroxy-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carboxylate (25 mg, 0.076 mmol) and DIPEA (0.053 ml, 0.306 mmol) 1,1,1-trifluoro-N-phenyl-N(trifluoromethyl)sulfonyl methanesulfonamide (82 mg, 0.229 mmol) was added to THF (3 ml). The resulting solution was stirred overnight at room temperature. 1-(bis(4-fluorophenyl)methyl)piperazine (22.04 mg, 0.076 mmol) and DIPEA (0.053 mL, 0.306 mmol) were added and the resulting mixture was stirred for 2 hours at room temperature. The reaction mixture was then concentrated in vacuo and the crude material was purified by preparative HPLC using acetonitrile-ammonium water acetate as the eluent. Homogeneous fractions were combined and evaporated in vacuo to give the title compound (4.4 mg, yield: 9.2%). Analytical LC-MS conditions: column: Waters XBridge C18, 2.1 mm x 50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). LC-MS results: 2.6 min, 597.0 (M+H)+. 1H NMR (500 MHz, DMSO-d6) δ 7.89 (d, J=8.8 Hz, 1H), 7.79 (d, J=9.2 Hz, 1H), 7.49 (dd, J=8.3, 5.7 Hz, 4H), 7.14 (t, J=8.8 Hz, 4H), 4.51 (s, 1H), 4.27 (q, J=7.3 Hz, 2H), 3.52 (s, 3H), 3.42 (br. s, 4H), 2.51 (br. s, 4H), 1.28 (t, J=7.0 Hz, 3H).

Example 2 6-Bromo-4-(4-(2-hydroxybenzyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile

To a solution of 6-bromo-1-methyl-2-oxo-4-(piperazin-1-yl)-1,2-dihydro-1,5-naphthyridine-3-carbonitrile bis(2,2,2-trifluoroacetate) ( 25 mg, 0.043 mmol) in DMF (1.5 ml) was added 2-hydroxybenzaldehyde (7.95 mg, 0.065 mmol). This reaction mixture was stirred for 1 hour at room temperature. Sodium cyanoborohydride (8.18 mg, 0.130 mmol) was added and the reaction mixture was stirred at room temperature for an additional 2 hours. Methanol was added and the resulting mixture was filtered and then fractionated by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 microns; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 60-100% B for 20 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Homogeneous fractions were combined and evaporated using a centrifugal evaporator to obtain the title compound (9.3 mg, yield: 44.8%). Analytical LC-MS conditions: column: Waters XBridge C18, 2.1 mm x 50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). LC-MS results: 1.9 min, 454.0 (M+H)+. 1H NMR (500 MHz, DMSO-d6) δ 8.00-7.94 (m, 1H), 7.93-7.88 (m, 1H), 7.19 (br. d, J=7.7 Hz, 1H), 7.13 (br. t, J =7.5 Hz, 1H), 6.89-6.66 (m, 2H), 3.88 (br. s, 4H), 3.70 (br. s, 2H), 3.52 (s, 3H), 2.71 (br. s, 4H).

Example 3. 4-(4-(bis(4-Fluorophenyl)methyl)piperazin-1-yl)-1-methyl-3-nitro-1,5-naphthyridin-2(1H)one

- 68 044500

4-(4-(bis(4-Fluorophenyl)methyl)piperazin-1-yl)-3-nitro-1,5-nαphthyridin-2(1H)-one (400 mg, 0.838 mmol) and potassium carbonate (463 mg , 3.35 mmol) was mixed with DMF (4189 µl) in a round bottom flask. Iodomethane (119 mg, 0.838 mmol) was added and the reaction mixture was heated at 50°C overnight. The reaction mixture was then cooled and diluted with water. A yellow solid (0.26 g, 63%) was collected by filtration and dried under high vacuum. ¹ H NMR (400 MHz, DMSO-de) 5 8.57 (dd, J=4.4, 1.2 Hz, 1H), 8.04 (dd, J=8.8, 1.2 Hz, 1H), 7.72 (dd, J=8.7, 4.3 Hz , 1H), 7.49 (dd, J=8.6, 5.6 Hz, 4H), 7.15 (t, J=8.8 Hz, 4H), 4.51 (s, 1H), 3.58 (s, 3H), 3.49 (br. s. , 4H). ¹³ C NMR (101 MHz, chloroform-d) δ 163.2, 160.7, 155.8, 148.3, 143.2, 137.9, 137.8, 136.3, 134.6, 132.3, 129.3, 129.2, 125.8, 122.4, 115.7, 115.5, 74.5, 51.9, 50.9, 29.3. Analytical LC-MS conditions: Column: Phenomenex Luna C18, 2.0x50 mm, particle size 3.0 µm; mobile phase A: (5:95) methanol:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) methanol:water with 0.1% trifluoroacetic acid; gradient: 0-100% B for 4 min, then maintaining 100% B for 0.75 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: 3.2 min, 492 (M+H).

Examples 5-7. 6-Bromo-4-{4-[(4-fluorophenyl)[2-(prop-2-yn-1-yloxy)phenyl]methyl]piperazin-1-yl}-1methyl-2-oxo-1,2- Dihydro-1,5-naphthyridine-3-carbonitrile

Potassium carbonate ( 23.15 mg, 0.167 mmol) and 1-((4-fluorophenyl)(2(prop-2-yn-1-yloxy)phenyl)methyl)piperazine, 2 TFA (55.1 mg, 0.126 mmol). This reaction mixture was stirred by shaking for 2 hours at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 35-100% B for 15 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 2nd sample injection: purity: 100.0%; retention time: 1.7; observed adducts: [M+H]; measured masses: 585.97. Analysis results for the 1st sample input: purity: 100.0%; retention time: 2.4; observed adducts: [M+H]; measured masses: 585.99. The title compound (29.2 mg) was isolated in 59.3% yield. This racemic compound was purified using chiral chromatography SFC (supercritical fluid chromatography).

The compound of Example 6 (first eluting isomer) and the compound of Example 7 (second eluting isomer) were isolated from the racemate using SFC-chiral chromatography. Analytical LC-MS was used to determine the final purity of the products.

Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B:

- 69 044500 (95:5) acetonitrile: water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration:

MS and UV (220 nm).

Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Example 6. Analysis results for the 1st sample injection: purity: 100.0%; measured weight: 585.99; retention time: 2.32 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 585.94; retention time: 1.66 min. Analysis results for the 1st sample input: purity: 100.0%; retention time: 2.32; observed adducts: [M+H]; measured masses: 585.99. Example 6 (7.4 mg) was isolated in 15% yield.

Example 7. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 585.97; retention time: 2.32 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 585.96; retention time: 1.66 min. Example 7 (7.3 mg) was isolated in 14.8% yield.

Examples 8-10. 6-Bromo-4-{4-[(4-fluorophenyl)(2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile

Br N

Potassium carbonate ( 93 mg, 0.670 mmol) and 2-((4-fluorophenyl)(piperazin1-yl)methyl)phenol, 2 TFA (86 mg, 0.167 mmol). This reaction mixture was stirred by shaking for 2 hours at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 60-100% B for 20 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured weight: 548.05; retention time: 2.24 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured weight: 548.05; retention time: 1.47. The title racemic compound (25.8 mg) was isolated in 28% yield. The racemic substance was further purified using SFC-chiral chromatography to obtain isomers.

The compound of Example 9 (first eluting isomer) and the compound of Example 10 (second eluting isomer) were isolated from the racemate using SFC-chiral chromatography. Analytical LC-MS was used to determine the final purity of the products.

Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0% V to 100% V

- 70 044500 for 3 minutes, then maintaining 100% B for 0.75 minutes; flow rate: 1 ml/min; registration:

MS and UV (220 nm).

Example 9. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 548.11; retention time: 1.78 min. Analysis results for the 2nd sample injection: purity: 94.5%; measured mass: 548.08; retention time: 2.66 min. Example 9 (11.1 mg) was 94% pure after isolation.

Example 10. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 548.08; retention time: 1.78 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 548.08; retention time: 2.66 min. The compound of Example 10 (11.8 mg) was 100% pure after isolation.

Example 11 7-dicarbonitrile

DMF solution (1 ml) 5-methyl-6-oxo-8-(piperazin-1-yl)-5,6-dihydro-1,5-naphthyridine-2,7dicarbonitrile, TFA (20 mg, 0.049 mmol), (4-fluorophenyl)boronic acid (6.85 mg, 0.049 mmol) and 2hydroxybenzaldehyde (5.98 mg, 0.049 mmol) were sealed in a microwave-safe tube and heated for 2 h at 150 °C. According to LC-MS analysis, some reaction product was present in the mixture, but the starting material (5-methyl-6oxo-8-(piperazin-1-yl)-5,6-dihydro-1,5-naphthyridine- 2,7-dicarbonitrile, TFU). Another portion of 2-hydroxybenzaldehyde (5.98 mg, 0.049 mmol) and (4-fluorophenyl)boronic acid (6.85 mg, 0.049 mmol) was added and the reaction mixture was heated for 2 hours at 150°C. LC-MS analysis indicated that approximately 20% of the starting material was converted to the desired product. This crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 38-78% B for 22 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 495.14; retention time: 1.41 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 98.6%; measured mass: 495.14; retention time: 2.06 min. The title compound (1.5 mg) was isolated in 6.2% yield.

Examples 12-14. 6-Bromo-4-{4-[(4-fluorophenyl)(2-methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile

To a DMF solution (1.5 ml) of 6-bromo-4-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (50 mg, 0.167 mmol) was added Huenig (0.15 ml, 0.84 mmol) and then 1-((4-71 044500 fluorophenyl)(2-methoxyphenyl)methyl)piperazine, 2 TFA (89 mg, 0.167 mmol) were added. This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-100% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 562.12; retention time: 2.46 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 562.12; retention time: 1.59 min. The title racemic compound (50.4 mg) was isolated in 53.7% yield. The racemic substance was further purified using SFC-chiral chromatography to obtain isomers.

The compound of Example 13 (first eluting isomer) and the compound of Example 14 (second eluting isomer) were isolated from the racemate using SFC-chiral chromatography. Analytical LC-MS was used to determine the final purity of the products.

Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Example 13. Analysis results for the 1st sample injection: purity: 100.0%; measured weight: 561.99; retention time: 2.4 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 562; retention time: 1.55 min. Example 13 (15.2 mg) was isolated in 16.2% yield.

Example 14. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 562; retention time: 2.4 min. Analysis results for the 2nd sample injection: purity: 99.1%; measured mass: 562; retention time: 1.55 min. Example 14 (15.3 mg) was isolated in 16.3% yield.

Examples 15-17. 6-Bromo-4-{4-[(4-fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl]piperazin-1-yl}-1methyl-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile

CH ₃

C/ oK solution of 6-bromo-4-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (50 mg, 0.167 mmol) in DMF (1.5 ml) potassium carbonate (93 mg, 0.670 mmol) was added and then 1-((4fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl)piperazine, 2 TFA (92 mg, 0.167 mmol) was added. This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete. The formation of a precipitate was observed. DMF (2 ml) was added followed by ethyl acetate and washed with water and brine. The resulting crude substance was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-100% B for 20 min, then

- 72 044500 maintaining 100% B for 4 minutes; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 98.6%; measured mass: 580.12; retention time: 2.49 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 580.05; retention time: 1.7 min. The title racemic compound (22.7 mg) was isolated in 23.4% yield. The racemic substance was further purified using SFC-chiral chromatography to obtain isomers.

The compound of Example 16 (first eluting isomer) and the compound of Example 17 (second eluting isomer) were isolated from the racemate using SFC-chiral chromatography. Analytical LC-MS was used to determine the final purity of the products. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Example 16. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 580.07; retention time: 2.49 min. Analysis results for the 2nd sample injection: purity: 99.4%; measured mass: 580.09; retention time: 1.66 min. Example 16 (7.5 mg) was isolated in 7.7% yield.

Example 17. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 580.1; retention time: 2.49 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 580.1; retention time: 1.66 min. Example 17 (7.4 mg) was isolated in 7.6% yield.

Examples 18-20. 8-{4-[(4-Fluorophenyl)(2-methoxyphenyl)methyl]piperazin-1-yl}-5-methyl-7-nitro-6oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile

Huenig's base ( 0.099 ml, 0.567 mmol) and then 1((4-fluorophenyl)(2-methoxyphenyl)methyl)piperazine, 2 TFA (59.9 mg, 0.113 mmol) was added. This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 47-87% B for 20 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 529.17; retention time: 2.38 min. Analysis conditions for the 2nd input image

- 73 044500 tsa: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 microns; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 529.15; retention time: 1.62 min. The title racemic compound (42.4 mg) was isolated in 70% yield. The racemic substance was further purified using SFC-chiral chromatography to obtain isomers.

The compound of Example 19 (first eluting isomer) and the compound of Example 20 (second eluting isomer) were isolated from the racemate using chiral SFC chromatography. Analytical LC-MS was used to determine the final purity of the products.

Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Example 19. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 529.16; retention time: 2.38 min. Analysis results for the 2nd sample injection: purity: 99.4%; measured mass: 529.18; retention time: 1.62 min. Example 19 (9 mg) was isolated in 29% yield.

Example 20. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 529.17; retention time: 2.38 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 529.15; retention time: 1.62 min. Example 20 (9.9 mg) was isolated in 33% yield.

Example 21 -1,5-naphthyridine-3-carbonitrile

To a solution of 6-bromo-4-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (50 mg, 0.167 mmol) in DMF (1.5 ml) was added carbonate potassium (93 mg, 0.670 mmol) and then 1-(6methoxy-2,3-dihydro-1H-inden-1-yl)piperazine, 2 TFA (77 mg, 0.167 mmol) were added. This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude substance was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 40-80% B for 22 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 98.0%; measured mass: 494.06; retention time: 1.35 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 97.9%; measured mass: 494.08; retention time: 2.09 min. The title compound (19.7 mg) was isolated in 59.5% yield.

Examples 22-24. 6-Bromo-4-{4-[1-(4-fluorophenyl)ethyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro1,5-naphthyridine-3-carbonitrile

- 74 044500

To a solution of 6-bromo-4-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (20 mg, 0.067 mmol) and 1-(1-(4-fluorophenyl )ethyl)piperazine (27.9 mg, 0.134 mmol) in DMF (1 ml) was added Hunig's base (0.035 ml, 0.201 mmol). This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete.

The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 42-82% B for 20 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 470.05; retention time: 2.11 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 470.05; retention time: 1.3 min. The title compound (19.8 mg) was isolated in 62.8% yield. This racemic substance was further purified using SFC-chiral chromatography to obtain isomers.

Example 23 (first eluting isomer) and Example 24 (second eluting isomer) were isolated from the racemate using SFC chiral chromatography. Analytical LC-MS was used to determine the final purity of the products.

Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Example 23. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 470.06; retention time: 2.11 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 470.05; retention time: 1.26 min. Example 23 (6.2 mg) was isolated in 19.7% yield.

Example 24. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 470.03; retention time: 2.11 min. Analysis results for the 2nd sample injection: purity: 98.5%; measured mass: 470.04; retention time: 1.26 min. Example 24 (6.3 mg) was isolated in 20% yield.

Examples 25-27. 6-Bromo-4-{4-[1-(4-fluorophenyl)propyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile

- 75 044500

Base was added to a solution of 6-bromo-4-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (15 mg, 0.050 mmol) in DMF (0.5 ml) Huenig (0.026 ml, 0.151 mmol) and 1-(1-(4fluorophenyl)propyl)piperazine (22.34 mg, 0.100 mmol). This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 41-81% B for 20 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 484.1; retention time: 1.4 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 484.06; retention time: 2.26 min. The title racemic compound (20.4 mg) was isolated in 84.2% yield. The racemic substance was further purified using SFC-chiral chromatography to obtain isomers.

The compound of Example 26 (first eluting isomer) and the compound of Example 27 (second eluting isomer) were isolated from the racemate using chiral SFC chromatography. Analytical LC-MS was used to determine the final purity of the products.

Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Example 26. Analysis results for the 1st sample injection: purity: 96.7%; measured mass: 483.96; retention time: 1.53 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured weight: 483.97; retention time: 2.19 min. Example 26 (7.6 mg) was isolated in 31.4% yield.

Example 27. Analysis results for the 1st sample injection: purity: 97.3%; measured mass: 483.96; retention time: 1.34 min. Analysis results for the 2nd sample injection: purity: 97.7%; measured mass: 483.97; retention time: 2.19 min. Example 27 (7.4 mg) was isolated in 30.6% yield.

Examples 28-30. 6-Bromo-4-{4-[2-(4-fluorophenyl)-2-hydroxyethyl]piperazin-1-yl}-1-methyl-2-oxo1,2-dihydro-1,5-naphthyridine-3-carbonitrile

HE (28)

Base was added to a solution of 6-bromo-4-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (15 mg, 0.050 mmol) in DMF (0.5 ml) Huenig (0.026 ml, 0.151 mmol) and a mixture of 1-(4-fluorophenyl)-2-(piperazin-1-yl)ethan-1-ol, TFA and 2-(4-fluorophenyl)-2-(piperazin-1-yl) ethanol, TFA (34.0 mg, 0.100 mmol). This reaction mixture was stirred by shaking for 1 hour at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 25

- 76 044500

70% B for 20 min, then maintaining 100% B for 6 min; flow rate: 20 ml/min. Fractions containing the secondary alcohol isomer were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 2nd sample injection: purity: 98.9%; retention time: 1.16; observed adducts: [M+H]; measured masses: 485.91. Analysis results for the 1st sample input: purity: 100.0%; retention time: 1.61; observed adducts: [M+H]; measured masses: 485.93. The title racemic compound (15.4 mg) was isolated in 62% yield. The racemic substance was further purified using SFC-chiral chromatography to obtain isomers. Isomer 6-bromo-4-{4-[1-(4fluorophenyl)-2-hydroxyethyl]piperazin-1 -yl} -1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3 - carbonitrile was also isolated.

The compound of Example 29 (first eluting isomer) and the compound of Example 30 (second eluting isomer) were isolated from the racemate using chiral SFC chromatography. Analytical LC-MS was used to determine the final purity of the products.

Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Example 29. Analysis results for the 1st sample injection: purity: 98.3%; measured mass: 486.06; retention time: 1.65 min. Analysis results for the 2nd sample injection: purity: 98.0%; measured mass: 486.05; retention time: 1.21 min. Example 29 (6.2 mg) was isolated in 25.5% yield.

Example 30. Analysis results for the 1st sample injection: purity: 98.2%; measured mass: 486.06; retention time: 1.65 min. Analysis results for the 2nd sample injection: purity: 97.9%; measured mass: 486.03; retention time: 1.21 min. Example 30 (5.3 mg) was isolated in 21.8% yield.

Example 31 naphthyridine-3-carbonitrile

Fractions containing the primary alcohol isomer were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 2nd sample injection: purity: 94.2%; retention time: 1.2; observed adducts: [M+H];

- 77 044500 measured masses: 485.93. Analysis results for the 1st sample injection: purity: 95.2%; retention time: 1.72; observed adducts: [M+H]; measured masses: 485.95. The title compound (3.5 mg) was isolated in 14.4% yield.

Examples 32-34. 8-{4-[1 -(4-Fluorophenyl)propyl]piperazin-1-yl}-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile

1-( 1-(4-fluorophenyl)propyl)piperazine (18.48 mg, 0.083 mmol) and then Huenig's base (0.040 ml, 0.227 mmol). This reaction mixture was stirred by shaking for 2 hours at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 40-80% B for 20 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 451.14; retention time: 1.33 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured weight: 451.15; retention time: 2.17 min. The title racemic compound (27.3 mg) was isolated in 79.7% yield. The racemic substance was further purified using SFC-chiral chromatography to obtain isomers.

The compound of Example 33 (first eluting isomer) and the compound of Example 34 (second eluting isomer) were isolated from the racemate using chiral SFC chromatography. Analytical LC-MS was used to determine the final purity of the products. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Example 33. Analysis results for the 1st sample injection: purity: 98.9%; measured mass: 451.11; retention time: 1.33 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured weight: 451.15; retention time: 2.17 min. Example 33 (7.7 mg) was isolated in 22.5% yield.

Example 34. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 451.13; retention time: 1.33 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 451.14; retention time: 2.17 min. Example 34 (8.4 mg) was isolated in 24.5% yield.

Example 35

- 78 044500

To a solution of 1-(cyclopropyl(4-fluorophenyl)methyl)piperazine, TFA (23.34 mg, 0.067 mmol) and 6-bromo4-chloro-1-methyl-2-oxo-1,2-dihydro-1,5- Naphthyridine-3-carbonitrile (20 mg, 0.067 mmol) in DMF (1 ml) was added with Hunig's base (0.012 ml, 0.067 mmol). This reaction mixture was stirred by shaking for 2 hours at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 43-83% B for 20 min, then maintain 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 496.13; retention time: 2.25 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 496.07; retention time: 1.39 min. The title compound (5.1 mg) was isolated in 15.3% yield.

Example 36 2-carbonitrile

A mixture of 5-methyl-7-nitro-6-oxo-8-(piperazin-1-yl)-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (20 mg, 0.064 mmol) and 2-(4 -fluorophenyl)oxirane (26.4 mg, 0.191 mmol) in ethanol (2 ml) was heated at reflux overnight. DMF (2 ml) was added and the reaction mixture was heated in a microwave oven at 110°C for 45 minutes. LC-MS analysis indicated approximately 50% conversion of the starting materials. The reaction mixture was heated at 120°C in a microwave oven for 1 hour. LC-MS analysis indicated approximately 80% conversion to the desired product. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 20-80% B for 22 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 453.11; retention time: 1.2 min. Analysis conditions for 2nd sample injection: column

- 79 044500 ka: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 microns; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 98.6%; measured mass: 453.13; retention time: 1.74 min. The title compound (3.2 mg) was isolated in 11.1% yield.

Example 37 2-carbonitrile

The second isomer was isolated during the synthesis of 8-{4-[2-(4-fluorophenyl)-2-hydroxyethyl]piperazin-1-yl}-5methyl-7-nitro-6-oxo-5,6-dihydro-1,5- naphthyridine-2-carbonitrile and purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 20-80% B for 22 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for 1st sample injection: Column: Waters XBridge

C18, 2.1 mmx50 mm, particle size 1.7 microns; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 96.8%; measured mass: 453.16; retention time: 1.16 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 453.1; retention time: 1.61 min. The title compound (1.2 mg) was isolated in 4.1% yield.

Example 38 1-Methyl-4-{4-[(naphthalene-1-yl)methyl]piperazin-1-yl}-3-nitro-1,2-dihydro-1,5-naphthyridin-2-one

To a solution of 4-(4-(naphthalene-1-ylmethyl)pinerazin-1-yl)-3-nitro-1,5-naphthyridin-2(1H)-one (20 mg, 0.048 mmol) in DMF (1 ml) sodium hydride (2.311 mg, 0.096 mmol) was added. This reaction mixture was stirred for 5 minutes at room temperature. Iodomethane (6.02 µl, 0.096 mmol) was added and the reaction mixture was stirred overnight at room temperature. According to LCMS analysis, the reaction was complete. The resulting crude substance was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 35-75% B for 15 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water

- 80 044500 with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 1st sample injection: purity: 100%; measured mass: 430.0; retention time: 2.5 min. Analysis results for the 2nd sample injection: purity: 94.7%; measured mass: 430.0; retention time: 1.5 min. The title compound (5.4 mg) was isolated in 26.2% yield.

Example 3 9. 6-Chloro-4-{4-[(4-fluorophenyl) [2-(prop-2-yn-1 -yloxy)phenyl]methyl] piperazin-1 -yl}-1 methyl-2-oxo -1,2-dihydro-1,5-naphthyridine-3-carbonitrile

DIPEA (0.066 mL, 0.378 mmol) and then 1-((4-fluorophenyl)(2-(prop-2-yn1-yloxy)phenyl)methyl)piperazine, TFA (60.7 mg, 0.139 mmol). This reaction mixture was stirred by shaking for 2 hours at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-90% B for 15 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured weight: 542.05; retention time: 1.54 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured weight: 542.05; retention time: 2.32 min. The title compound (33.8 mg) was isolated in 49.5% yield.

Example 40 8-(4-(bis(4-Fluorophenyl)methyl)piperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5naphthyridine-2-carbonitrile

DMF was bubbled with nitrogen for 1 hour. Zinc (0.95 mg, 0.015 mmol), bromo(tri-tert-butylphosphine)palladium (I) dimer (9.96 mg, 0.013 mmol) and 4- (4-(bis(4fluorophenyl)methyl)niperazin-1-yl)-6-bromo-1-methyl-3-nitro-1,5-naphthyridin-2(1H)-one (21.38 mg, 0.037 mmol) . Bubbled DMF (0.3 ml) was added and the mixture was sealed under nitrogen and immersed in a 50° C. oil bath for 15 minutes. Dicyanozinc (2.86 mg, 0.024 mmol) was added. The reaction mixture was sealed under a nitrogen atmosphere and immersed in an oil bath at 50°C for 3 hours. According to LC-MS analysis, the reaction was complete. The resulting crude substance

- 81 044500 was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-90% B for 15 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. The title compound (11.4 mg) was isolated in 59.7% yield.

Alternative synthesis. A solution of 8-chloro-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2carbonitrile (750 mg, 2.83 mmol) in DMF (6 ml) was mixed with 1-( bis(4-fluorophenyl)methyl)piperazine (899 mg, 3.12 mmol) and then Huenig's base (0.990 ml, 5.67 mmol) was added. This reaction mixture was stirred overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was filtered and purified by preparative HPLC using aqueous acetonitrile with ammonium acetate as a buffer to give a yellow solid (1.02 g). To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 1st sample injection: purity: 100%; measured mass: 517.0; retention time: 2.4 min. Analysis results for the 2nd sample injection: purity: 98.4%; measured mass: 517.0; retention time: 1.7 min. 1H NMR (500 MHz, chloroform-d) δ 7.88 (d, J=8.7 Hz, 1H), 7.76 (d, J=8.9 Hz, 1H), 7.40 (dd, J=8.5, 5.5 Hz, 4H), 7.02 (t, J=8.7 Hz, 4H), 4.34 (s, 1H), 3.68 (s, 3H), 3.62-3.55 (m, 4H), 2.64 (br. s, 4H). ¹³ C NMR (126 MHz, chloroform-d) δ 163.0, 161.0, 155.4, 147.0, 138.0, 137.7, 137.7, 135.9, 132.4, 129.5, 129.2, 129.2, 126.0, 123.1, 116.5, 115.8, 115.6, 74.3, 51.6, 51.2, 29.7.

Example 41 8-(4-Benzhydrylpiperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridin2-carbonitrile

DMF was bubbled with nitrogen for 1 hour. Zinc (1.2 mg, 0.018 mmol), bromo(tri-tert-butylphosphine)palladium(I) dimer (16 mg, 0.021 mmol) and 4-(4 -benzhydrylpiperazin-1-yl)-6-bromo-1-methyl-3-nitro-1,5-naphthyridin-2(1H)-one (22.82 mg, 0.043 mmol). Bubbled DMF (0.3 mL) was added and the mixture was capped under nitrogen. The test tube was immersed for 15 minutes in an oil bath at 50°C. Dicyanozinc (3.7 mg, 0.032 mmol) was added. The tube was hermetically sealed under a nitrogen atmosphere and immersed for 3 hours in an oil bath at 50°C. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-100% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LCMS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 1st sample injection: purity: 100%; measured

- 82 044500 weight: 481.0; retention time: 2.4 min. Analysis results for the 2nd sample injection: purity: 100%;

measured mass: 481.0; retention time: 1.6 min. The title compound (13 mg) was isolated in 62.9% yield.

Example 42 carbonitrile

DMF was bubbled with nitrogen for 1 hour. Zinc (0.95 mg, 0.015 mmol), bromo(tri-tert-butylphosphine)palladium(I) dimer (9.96 mg, 0.013 mmol) and 6- bromo-4-(4((2-hydroxyphenyl)(phenyl)methyl)piperazin-1-yl)-1-methyl-3-nitro-1,5-naphthyridin-2(1H)-one (22.02 mg, 0.040 mmol). DMF (0.3 ml) was added. The tube was sealed under a nitrogen atmosphere and immersed for 15 min in an oil bath at 50°C. Dicyanozinc (2.86 mg, 0.024 mmol) was added. The tube was hermetically sealed under a nitrogen atmosphere and immersed for 3 hours in an oil bath at 50°C. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 40-90% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 1st sample injection: purity: 95.4%; measured mass: 550.0; retention time: 2.3 min. Analysis results for the 2nd sample injection: purity: 96.0%; measured mass: 550.0; retention time: 1.6 min. The title compound (5 mg) was isolated in 25.2% yield.

Examples 43-45. 8-(4-(1-(4-Fluorophenyl)ethyl)piperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6-dihydro1,5-naphthyridine-2-carbonitrile

To a solution of 8-chloro-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (20 mg, 0.060 mmol) and 1-(1-phenylethyl)piperazine ( 50.6 mg, 0.121 mmol) to DMF (1 ml) was added Huenig's base (0.032 ml, 0.181 mmol). This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 37-77% B for 20 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonite-83 044500 ril:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 437.11; retention time: 2.05 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 437.11; retention time: 1.23 min. The title racemic compound (18 mg) was isolated in 68.7% yield. The racemic substance was further purified using SFC-chiral chromatography to obtain isomers.

The compound of Example 44 (first eluting isomer) and the compound of Example 45 (second eluting isomer) were isolated from the racemate using SFC-chiral chromatography. Analytical LC-MS was used to determine the final purity of the products.

Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Example 44. Analysis results for the 1st sample injection: purity: 97.5%; measured mass: 437.14; retention time: 1.24 min. Analysis results for the 2nd sample injection: purity: 98.4%; measured mass: 437.11; retention time: 2.06 min. Example 44 (6.7 mg) was isolated in 25.6% yield.

Example 45. Analysis results for the 1st sample injection: purity: 97.5%; measured mass: 437.14; retention time: 1.24 min. Analysis results for the 2nd sample injection: purity: 98.4%; measured mass: 437.11; retention time: 2.06 min. Example 45 (7.2 mg) was isolated in 27.5% yield.

Example 46 1,5-naphthyridine-2-carbonitrile

To a solution of 8-chloro-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (20 mg, 0.060 mmol) and 1-(6-methoxy-2, 3-dihydro-1H-inden-1-ul)piperazine (55.7 mg, 0.121 mmol) in DMF (1 ml) was added Huenig's base (0.032 ml, 0.181 mmol). This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude substance was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 41-81% B for 20 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 99.3%; measured mass: 461.15; retention time: 2.03 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for 2nd input

- 84 044500 sample: purity: 100.0%; measured mass: 461.15; retention time: 1.29 min. The title compound (16.1 mg) was isolated in 58.3% yield.

Example 47 8-(4-(2-Hydroxy-1-phenylethyl)piperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6-dihydro1,5-naphthyridine-2-carbonitrile

To 5-methyl-7-nitro-6-oxo-8-(piperazin-1-yl)-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (20 mg, 0.064 mmol) in ethanol (2 ml ) 2-phenyloxirane (15.29 mg, 0.127 mmol) was added. This reaction mixture was heated at reflux overnight. LC-MS analysis indicated the absence of the desired product. DMF (2 ml) was added and the reaction mixture was heated in a microwave oven at 100°C for 30 minutes. LC-MS analysis indicated approximately 50% conversion of the starting materials. The reaction mixture was heated for 45 min at 120°C. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 25-75% B for 25 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 435.14; retention time: 1.68 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 435.16; retention time: 1.16 min. The title compound (7.1 mg) was isolated in 25.5% yield.

Example 48 8-(4-(2-Hydroxy-2-phenylethyl)piperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6-dihydro1,5-naphthyridine-2-carbonitrile

The second isomer was isolated during the synthesis of 8-[4-(2-hydroxy-2-phenylethyl)piperazin-1-yl]-5-methyl-7nitro-6-oxo-5,6-dihydro-1,5-naphthyridin-2- carbonitrile. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for 1st sample injection: Column: Waters XBridge

C18, 2.1 mmx50 mm, particle size 1.7 microns; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 435.12; retention time: 1.56 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 435.12; retention time: 1.13

- 85 044500 min. The title compound (2.2 mg) was isolated in 8% yield.

Examples 49-51. 8-(4-(Cyclopropyl(4-fluorophenyl)methyl)piperazin-1-yl)-5-methyl-7-nitro-6-oxo5,6-dihydro-1,5-naphthyridine-2-carbonitrile

To a solution of 1-(cyclopropyl(4-fluorophenyl)methyl)piperazine TFA (26.3 mg, 0.076 mmol) and 8-chloro-5methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine -2-carbonitrile (20 mg, 0.076 mmol) in DMF (1 ml) was added Hunig's base (0.013 ml, 0.076 mmol). This reaction mixture was stirred by shaking for 2 hours at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 41-81% B for 20 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. The resulting substance was further purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 32-72% B for 25 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 463.12; retention time: 1.35 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 463.12; retention time: 2.18 min. The title racemic compound (13.7 mg) was isolated in 39% yield. The racemic substance was further purified using SFC-chiral chromatography to obtain isomers.

The compound of Example 50 (first eluting isomer) and the compound of Example 51 (second eluting isomer) were isolated from the racemate using SFC-chiral chromatography. Analytical LC-MS was used to determine the final purity of the products.

Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Example 50. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 463.03; retention time: 1.35 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 463.03; retention time: 2.15 min. Example 50 (6.7 mg) was isolated in 19.1% yield.

Example 51. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 463.02; retention time: 1.35 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 463.02; retention time: 2.16 min. Example 51 (6.5 mg) was isolated in 18.5% yield.

Example 52 4-(4-Benzhydrylpiperazin-1-yl)-6-bromo-1-methyl-3-nitro-1,5-naphthyridin-2(1H)-one

- 86 044500

To a solution of 6-bromo-4-chloro-1-methyl-3-nitro-1,5-naphthyridin-2(1H)-one (20 mg, 0.063 mmol) in DMF (1 ml) was added DIPEA (0.033 ml, 0.188 mmol) and 1-benzhydrylpiperazine (15.85 mg, 0.063 mmol). This reaction mixture was stirred by shaking at 35°C overnight. According to LC-MS analysis, the reaction was complete. The resulting clear yellow solution was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-90% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 1st sample injection: purity: 100%; measured mass: 534.0; retention time: 2.6 min. Analysis results for the 2nd sample injection: purity: 83.0%; measured mass: 534.0; retention time: 1.6 min. The title compound (9.8 mg) was isolated in 29.1% yield.

Example 53 4-(4-(bis(4-Fluorophenyl)methyl)piperazin-1-yl)-6-bromo-1-methyl-3-nitro-1,5-naphthyridin-2(1H)-one

To a solution of 6-bromo-4-chloro-1-methyl-3-nitro-1,5-naphthyridin-2(1H)-one (20 mg, 0.063 mmol) in DMF (1 ml), DIPEA (0.033 ml, 0.188 mmol) and 1-(bis(4-fluorophenyl)methyl)piperazine (18.11 mg, 0.063 mmol). This reaction mixture was stirred by shaking at 35°C overnight. According to LC-MS analysis, the reaction was complete. The resulting clear yellow solution was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 55-95% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LCMS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0

- 87 044500 ml/min; registration: UV, 220 nm. Analysis results for the 1st sample injection: purity: 100%; measured mass: 570.0; retention time: 2.6 min. Analysis results for the 2nd sample injection: purity: 100.0%;

measured mass: 570.0; retention time: 1.8 min. The title compound (16.4 mg) was isolated in 45.6% yield.

Example 54 2( 1 H)-one

To a solution of 6-bromo-4-chloro-1-methyl-3-nitro-1,5-naphthyridin-2(1H)-one (20 mg, 0.063 mmol) in DMF (1 ml), DIPEA (0.033 ml, 0.188 mmol) and 1-ethyl-4-(piperazin-1-ylmethyl)-1 H-indole, 2 TFA (29.6 mg, 0.063 mmol). This reaction mixture was stirred by shaking at 35°C overnight. According to LC-MS analysis, the reaction was complete. The resulting clear yellow solution was purified using reverse phase HPLC for purification. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-90% B for 15 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LCMS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 1st sample injection: purity: 100%; measured mass: 524.9; retention time: 2.3 min. Analysis results for the 2nd sample injection: purity: 99.1%; measured mass: 524.9; retention time: 1.5 min. The title compound (14.9 mg) was isolated in 45% yield.

Example 5 5. 6-Bromo-1-methyl-4-(4-(naphthalene-1 -ylmethyl)piperazin-1 -yl)-3-nitro-1,5-naphthyridin2(1H)-one

1-(Naphthalene-1 -ylmethyl)piperazine (17.05 mg, 0.075 mmol) and then DIPEA (0.033 ml, 0.188 mmol) was added. This reaction mixture was stirred by shaking at 35°C overnight. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 55-95% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LCMS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, size

- 88 044500 particles 1.7 microns; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 1st sample injection: purity: 100%; measured mass: 508.0; retention time: 2.5 min. Analysis results for the 2nd sample injection: purity: 100%; measured mass: 508.0; retention time: 1.5 min. The title compound (18.3 mg) was isolated in 57.1% yield.

Examples 56-58. 6-Bromo-4-(4-((4-fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl)piperazin-1-yl)-1-methyl-3-nitro-1,5-naphthyridine-2( 1 N)-he

1-((4- fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl)piperazine, TFA (102 mg, 0.235 mmol) and DIPEA (0.082 ml, 0.471 mmol). This reaction mixture was stirred by shaking at 35°C overnight. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; gradient: 18-58% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 1st sample injection: purity: 99.0%; retention time: 2.57; observed adducts: [M+H]; measured masses: 599.97. Analysis results for the 2nd sample injection: purity: 95.5%; retention time: 1.69; observed adducts: [M+H]; measured masses: 600. The title racemic compound (52.5 mg) was isolated in 55.7% yield. The racemic substance was further purified using SFC chiral chromatography to obtain isomers.

Example 57 (first eluting isomer) and Example 58 (second eluting isomer) were isolated from the racemate using SFC chiral chromatography. To determine the final purity of the products, samples were analyzed in duplicate using analytical LC-MS.

Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Example 57. Analysis results for the 1st sample injection: purity: 100.0%; retention time: 2.71;

- 89 044500 observed adducts: [M+H]; measured masses:

599.95. Analysis results for the 2nd sample injection: purity: 100.0%; retention time: 1.76; observed adducts: [M+H]; measured masses: 599.98. Example 57 (13.2 mg) was isolated in 14% yield.

Example 58. Analysis results for the 1st sample injection: purity: 100.0%; retention time: 2.71; observed adducts: [M+H]; measured masses: 599.96. Analysis results for the 2nd sample injection: purity: 99.1%; retention time: 1.75; observed adducts: [M+H]; measured masses: 599.98. Example 58 (13.7 mg) was isolated in 14.5% yield.

Examples 59-61. 6-Bromo-4-(4-((4-fluorophenyl)(2-methoxy-6-methylphenyl)methyl)piperazin-1-yl)-1-methyl-3-nitro-1,5-naphthyridine-2(1 N)-he

CH ₃

A

V o' ^СНз

To a solution of 6-bromo-4-(4-((4-fluorophenyl)(2-hydroxy-6-methylphenyl)methyl)piperazin-1 -yl)-1 -methyl3-nitro-1,5-naphthyridin-2(1H )-one (30 mg, 0.052 mmol) in DMF (2 ml) was added cesium carbonate (50.3 mg, 0.155 mmol). This reaction mixture was stirred for 20 minutes at room temperature and methyl iodide (9.66 μl, 0.155 mmol) was added. The reaction mixture was stirred for 1 hour at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 55-95% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 2nd sample injection: purity: 100.0%; retention time: 1.67; observed adducts: [M+H]; measured masses: 595.95. Analysis results for the 1st sample input: purity: 100.0%; retention time: 2.73; observed adducts: [M+H]; measured masses: 595.96. The title racemic compound (17.1 mg) was isolated in 55.1% yield. The racemic substance was further purified using SFC-chiral chromatography to obtain isomers.

The compound of Example 60 (first eluting isomer) and the compound of Example 61 (second eluting isomer) were isolated from the racemate using SFC-chiral chromatography. To determine the final purity of the products, samples were analyzed in duplicate using analytical LC-MS.

Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Example 60. Analysis results for the 1st sample injection: purity: 100.0%; retention time: 2.65; observed adducts: [M+H]; measured masses: 595.97. Analysis results for the 2nd sample injection: purity: 100.0%; retention time: 1.67; observed adducts: [M+H]; measured masses: 595.96. Example 60 (5.4 mg) was isolated in 17.4% yield.

- 90 044500

Example 61. Analysis results for the 1st sample injection: purity: 100.0%; retention time: 2.65;

observed adducts: [M+H]; measured masses: 595.96. Example 61 (5.1 mg) was isolated in 16.4% yield.

Example 62. tert-Butyl (8-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6dihydro-1,5-naphthyridin- 2-yl)carbamate

A mixture of tris(dibenzylideneacetone)dipalladium (0) (9.38 mg, 10.24 µmol), 4,5-bis(diphenylphosphino)9,9-dimethylxanthene (17.77 mg, 0.031 mmol), tert-butylcarbamate (39, 0 mg, 0.333 mmol), cesium carbonate (125 mg, 0.384 mmol) and 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-6-bromo-1-methyl-3-nitro- 1,5naphthyridin-2(1H)-one (146 mg, 0.256 mmol) was placed under argon. Dioxane (2560 µl) and H ₂ O (5.53 µl, 0.307 mmol) were added. The reaction mixture was sealed under argon and heated at 100°C overnight. According to LC-MS analysis, the reaction was complete. The solvent was removed. The residue was purified on a BIOTAGE™ MPLC system using hexanes:ethyl acetate (1:1) and a silica gel column (24 g). Fractions corresponding to the title compound were collected to obtain 160 mg of product as a light yellow film. A 10 mg sample was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 60-100% B for 15 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 1st sample injection: purity: 98.7%; retention time: 2.55; observed adducts: [M+H]; measured masses: 607.03. Analysis results for the 2nd sample injection: purity: 97.0%; retention time: 1.79; observed adducts: [M+H]; measured masses: 607.04. The title compound (3.2 mg) was isolated in 32% yield.

Example 63 6-Amino-4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-1-methyl-3-nitro-1,5-naphthyridin-2(1H)-one

A solution of tert-butyl (8-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine- 2-yl)carbamate (150 mg, 0.247 mmol) in dichloromethane (3 ml) was mixed with TFA (0.5 ml, 6.49 mmol). This reaction mixture was stirred for 2 hours at room temperature. According to LC-MS analysis, the reaction was complete. The solvent was removed to obtain a yellow solid. A 6 mg sample was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 35-75% B for 20 min, then maintaining 100% B for 4 min; speed

- 91 044500 flow: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 1st sample injection: purity: 98.8%; retention time: 2.16; observed adducts: [M+H]; measured masses: 507.1. Analysis results for the 2nd sample injection: purity: 100.0%; retention time: 1.7; observed adducts: [M+H]; measured masses: 507.08. The title compound (3.4 mg) was isolated in 55.9% yield.

Example 64 -1,5-naphthyridine-3-carbonitrile

To 2-(difluoromethyl)benzaldehyde (16.05 mg, 0.103 mmol) was added polymeric N,N,N-trimethyl1-(p-tolyl)methanamonium cyanoborohydride (4.1 mmol/g) (35 mg, 0.144 mmol). A mixture of 6-bromo2-oxo-4-(piperazin-1 -yl)-1 -(prop-2-yn-1 -yl)-1,2-dihydro-1,5-naphthyridin-3-carbonitrile TFU ( 25 mg, 0.051 mmol) in dichloromethane (2 ml) and acetic acid (0.250 ml). This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 45-85% B for 18 min, then maintain 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 512.04; retention time: 2.39 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured weight: 512.05; retention time: 1.68 min. The title compound (16.6 mg) was isolated in 63.5% yield.

Example 65 naphthyridine-3-carbonitrile

- 92 044500

Solution of 2-hydroxybenzaldehyde (22.60 mg, 0.185 mmol), 6-bromo-2-oxo-4-(piperazin-1-yl)-1(prop-2-yn-1-yl)-1,2-dihydro -1,5-naphthyridine-3-carbonitrile TFA (60 mg, 0.123 mmol) and sodium cyanoborohydride (15.51 mg, 0.247 mmol) in dichloromethane (2 ml) were stirred by shaking overnight at room temperature. LC-MS analysis indicated approximately 30% conversion of the starting materials. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 38-78% B for 19 min, then maintain 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured weight: 477.99; retention time: 1.97 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured weight: 477.98; retention time: 1.18 min. The title compound (16.7 mg) was isolated in 28.4% yield.

Example 66 1,5-naphthyridine-3-carbonitrile

A solution of 2-hydroxy-4-methylbenzaldehyde (7.56 mg, 0.056 mmol) and 6-bromo-2-oxo-4-(piperazin-1yl)-1-(prop-2-yn-1-yl)-1, 2-dihydro-1,5-naphthyridine-3-carbonitrile TFA (18 mg, 0.037 mmol) in DMF (2 ml) was stirred by shaking for 1 hour at room temperature. Sodium cyanoborohydride (4.65 mg, 0.074 mmol) was added. This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 47-87% B for 19 min, then maintain 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LCMS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 492.02; retention time: 2.19 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 492.05; retention time: 1.33 min. The title compound (8 mg) was isolated in 43.9% yield.

Example 67 1,5-naphthyridine-3-carbonitrile

- 93 044500

A solution of 4-fluoro-2-hydroxybenzaldehyde (7.78 mg, 0.056 mmol) and 6-bromo-2-oxo-4-(piperazin-1yl)-1-(prop-2-yn-1-yl)-1, 2-dihydro-1,5-naphthyridine-3-carbonitrile TFA (18 mg, 0.037 mmol) in DMF (2 ml) was stirred by shaking for 1 hour at room temperature. Sodium cyanoborohydride (4.65 mg, 0.074 mmol) was added. This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 42-82% B for 19 min, then maintain 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LCMS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured weight: 496.05; retention time: 1.28 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 496.04; retention time: 2.11 min. The title compound (9.8 mg) was isolated in 53.4% yield.

Example 68 1, 5-naphthyridine-3-carbonitrile

A solution of 4-fluoro-2-methoxybenzaldehyde (8.56 mg, 0.056 mmol) and 6-bromo-2-oxo-4-(piperazin-1-yl)1-(prop-2-yn-1-yl)-1 ,2-dihydro-1,5-naphthyridine-3-carbonitrile TFA (18 mg, 0.037 mmol) in DMF (2 ml) was stirred by shaking for 1 hour at room temperature. Sodium cyanoborohydride (4.65 mg, 0.074 mmol) was added. This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 42-82% B for 20 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV

- 94 044500 (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 510.09; retention time: 2.11 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 510.07; retention time: 1.41 min. The title compound (4.9 mg) was isolated in 25.9% yield.

Example 69 -dihydro-1,5-naphthyridine-3-carbonitrile

A solution of 2-hydroxy-4,6-dimethylbenzaldehyde (11.58 mg, 0.077 mmol) and 6-bromo-2-oxo-4(piperazin-1-yl)-1-(prop-2-yn-1-yl) -1,2-dihydro-1,5-naphthyridine-3-carbonitrile TFA (25 mg, 0.051 mmol) in DMF (2 ml) was stirred by shaking for 1 hour at room temperature. Sodium cyanoborohydride (6.46 mg, 0.103 mmol) was added. This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 35-83% B for 20 min, then maintain 100% B for 6 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 506.04; retention time: 2.27 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 506.08; retention time: 1.46 min. The title compound (9.9 mg) was isolated in 38.3% yield.

Example 70 He

CH ₃

XXX Br^N^|^NO ₂ O OH N

OXJl (70)

(4-fluorophenyl)boronic acid (13.99 mg, 0.100 mmol), salicylaldehyde (12.21 mg, 0.100 mmol) and 6-bromo-1-methyl-3-nitro-4-(piperazine) were added to a microwavable tube -1-yl)-1,5naphthyridin-2(1H)-one (36.8 mg, 0.1 mmol). The reaction vessel was sealed and heated using microwave radiation for 2 hours at 150°C in a Biotage™ Initiator microwave oven. According to LC-MS analysis, the reaction was complete. The resulting crude substance was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm,

- 95 044500 particle size 5 microns; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 45-85% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 2nd sample injection: purity: 98.0%; retention time: 1.59; observed adducts: [M+H]; measured masses: 567.99. Analysis results for the 1st sample input: purity: 100.0%; retention time: 2.42; observed adducts: [M+H]; measured masses: 568.02. The title compound (3 mg) was isolated in 5.3% yield.

Example 71 1 N)-on

(2-fluoro-4-methylphenyl)boronic acid (15.39 mg, 0.100 mmol), salicylaldehyde (12.21 mg, 0.100 mmol) and 6-bromo-1-methyl-3-nitro- 4-(piperazin-1-yl)-1,5naphthyridin-2(1H)-one (36.8 mg, 0.1 mmol). The reaction vessel was sealed and heated using microwave radiation for 2 hours at 150°C in a Biotage™ Initiator microwave oven. According to LC-MS analysis, the reaction was complete. This crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-100% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 2nd sample injection: purity: 100.0%; retention time: 1.69, 1.72; observed adducts: [M+H], measured masses: 582, 582. Analytical results for 1st sample injection: purity: 98.1%; retention time: 2.51, 2.56; observed adducts: [M+H]; measured masses: 582. The title compound (2.1 mg) was isolated in 3.6% yield.

Example 72 -He

- 96 044500

The compound of Example 72 was prepared according to the general procedure used for the synthesis of 6-bromo-4-{4-[(2-fluoro-4-methylphenyl)(2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl- 3-nitro-1,2-dihydro-1,5-naphthyridin-2-one. The resulting crude material was purified by preparative LCMS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-100% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 1st sample input: purity: 100.0%; retention time: 2.59; observed adducts: [M+H]; measured masses: 577.98. Analysis results for the 2nd sample injection: purity: 98.3%; retention time: 1.72; observed adducts: [M+H]; measured masses: 577.99. The title compound (2.1 mg) was isolated in 3.6% yield.

Example 7 3. 6-Bromo-4-(4-((2-hydroxyphenyl)(ortho-tolyl)methyl)piperazin-1 -yl)-1 -methyl-3-nitro1,5-naphthyridine-2(1H) -He

The compound of Example 73 was prepared according to the general procedure used for the synthesis of 6-bromo-4-{4-[(2-fluoro-4-methylphenyl)(2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl- 3-nitro-1,2-dihydro-1,5-naphthyridin-2-one. The resulting crude material was purified by preparative LCMS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-90% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 1st sample input: purity: 100.0%; retention time: 2.48; observed adducts: [M+H]; measured masses: 564.01. Analysis results for the 2nd sample injection: purity: 95.2%; retention time: 1.59; observed adducts: [M+H]; measured masses: 564.01. Stated in the title

- 97 044500 compound (6.2 mg) was isolated in 15% yield.

Examples 74-76. 6-Bromo-4-(4-((3-fluoro-2-hydroxyphenyl)(4-fluorophenyl)methyl)piperazin-1-yl)-1-methyl-3-nitro-1,5-naphthyridine-2(1 N)-he

2-Fluoro-6- ((4-fluorophenyl)(piperazin-1-yl)methyl)phenol, TFA (99 mg, 0.235 mmol) and DIPEA (0.082 ml, 0.471 mmol). This reaction mixture was stirred by shaking at 35°C overnight. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 5090% B for 20 min, then maintain 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 2nd sample injection: purity: 92.2%; retention time: 1.73; observed adducts: [M+H]; measured masses: 585.98. Analysis results for the 1st sample input: purity: 100.0%; retention time: 2.42; observed adducts: [M+H]; measured masses: 585.95. The title compound (60.7 mg) was isolated in 65.9% yield. The racemic substance was further purified using SFC-chiral chromatography to obtain isomers.

The compound of Example 75 (first eluting isomer) and the compound of Example 76 (second eluting isomer) were isolated from the racemate using chiral SFC chromatography. To determine the final purity of the products, samples were analyzed in duplicate using analytical LC-MS.

Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Example 75. Analysis results for the 1st sample injection: purity: 100.0%; retention time: 2.31; observed adducts: [M+H]; measured masses: 585.97. Analysis results for the 2nd sample injection: purity: 100.0%; retention time: 1.7; observed adducts: [M+H]; measured masses: 585.98. Example 75 (21 mg) was isolated in 22.8% yield.

Example 76. Analysis results for the 1st sample injection: purity: 100.0%; retention time: 2.31; observed adducts: [M+H]; measured masses: 585.97. Analysis results for the 2nd sample injection: purity: 100.0%; retention time: 1.67, 1.7; observed adducts: [M+H], [M+H]; measured masses: 585.98, 585.98. Example 76 (21.8 mg) was isolated in 23.7% yield.

Examples 77-79. 6-Bromo-4-(4-((2-hydroxyphenyl)(phenyl)methyl)piperazin-1-yl)-1-methyl-3-nitro1,5-naphthyridin-2(1H)-one

- 98 044500

2-(phenyl(piperazine) -1-yl)methyl)phenol, TFA (90 mg, 0.235 mmol) and DIPEA (0.082 ml, 0.471 mmol). This reaction mixture was stirred by shaking at 35°C overnight. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 45-95% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LCMS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. LC-MS results (acetonitrile TFA)-2: purity: 96.3%; retention time: 1.65; observed adducts: [M+H]; measured masses: 550. LC-MS results (acetonitrile ammonium acetate)-2: purity: 100.0%; retention time: 2.34; observed adducts: [M+H]; measured masses: 550. The title racemic compound (27 mg) was isolated in 31% yield. The racemic substance was further purified using SFC-chiral chromatography to obtain isomers.

The compound of Example 78 (first eluting isomer) and the compound of Example 79 (second eluting isomer) were isolated from the racemate using SFC-chiral chromatography. To determine the final purity of the products, samples were analyzed twice using analytical LC-MS.

Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Example 78 LC-MS results (acetonitrile TFA)-2: purity: 96.3%; retention time: 1.65; observed adducts: [M+H]; measured masses: 550; LC-MS results (acetonitrile ammonium acetate)-2: purity: 100.0%; retention time: 2.34; observed adducts: [M+H]; measured masses: 550. The compound of Example 78 (11.4 mg) was isolated in 13.2% yield.

Example 79 LC-MS results (acetonitrile TFA)-2: purity: 95.4%; retention time: 1.63; observed adducts: [M+H]; measured masses: 550.04; LC-MS results (acetonitrile ammonium acetate)-2: purity: 96.2%; retention time: 2.35; observed adducts: [M+H]; measured masses: 550.01. Example 79 (11.3 mg) was isolated in 13.1% yield.

Examples 80-82. 6-Bromo-4-(4-((4-fluoro-2-hydroxyphenyl)(4-fluorophenyl)methyl)piperazin-1-yl)-1-methyl-3-nitro-1,5-naphthyridine-2( 1 N)-he

- 99 044500

5-Fluoro-2- ((4-fluorophenyl)(piperazin-1-yl)methyl)phenol, TFA (99 mg, 0.235 mmol) and DIPEA (0.082 ml, 0.471 mmol). This reaction mixture was stirred by shaking at 35°C overnight. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 4585% B for 20 min, then maintain 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 1st sample injection: purity: 93.2%; retention time: 2.43; observed adducts: [M+H]; measured masses: 585.96. Analysis results for the 2nd sample injection: purity: 91.7%; retention time: 1.7; observed adducts: [M+H]; measured masses: 585.95. The title racemic compound (17 mg) was isolated in 19% yield. The racemic substance was further purified using SFC-chiral chromatography to obtain isomers.

The compound of Example 81 (first eluting isomer) and the compound of Example 82 (second eluting isomer) were isolated from the racemate using SFC-chiral chromatography. To determine the final purity of the products, samples were analyzed twice using analytical LC-MS.

Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Example 81. Analysis results for the 1st sample injection: purity: 93.2%; retention time: 2.43; observed adducts: [M+H]; measured masses: 585.96. Analysis results for the 2nd sample injection: purity: 91.7%; retention time: 1.7; observed adducts: [M+H]; measured masses: 585.95. Example 81 (7.2 mg) was isolated in 7.8% yield.

Example 82. Analysis results for the 1st sample injection: purity: 90.8%; retention time: 2.43; observed adducts: [M+H]; measured masses: 585.92. Analysis results for the 2nd sample injection: purity: 95.9%; retention time: 1.7; observed adducts: [M+H]; measured masses: 585.9. Example 82 (5.3 mg) was isolated in 5.8% yield.

Examples 83-85. 6-Bromo-4-(4-((4-fluorophenyl)(2-hydroxy-3-methylphenyl)methyl)piperazin-1-yl)-1methyl-3-nitro-1,5-naphthyridine-2(1H)- He

- 100 044500

2-((4- fluorophenyl)(piperazin-1-yl)methyl)-6-methylphenol (70.7 mg, 0.235 mmol) and then DIPEA (0.027 ml, 0.157 mmol) was added. This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-90% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 2nd sample injection: purity: 96.0%; retention time: 1.85; observed adducts: [M+H]; measured masses: 582. Analysis results for the 1st sample injection: purity: 97.8%; retention time: 2.53, 2.6; observed adducts: [M+H]; measured masses: 581.98. The title racemic compound (15.7 mg) was isolated in 17.2% yield. The racemic substance was further purified using SFC-chiral chromatography to obtain isomers.

The compound of Example 84 (first eluting isomer) and the compound of Example 85 (second eluting isomer) were isolated from the racemate using SFC-chiral chromatography. To determine the final purity of the products, samples were analyzed twice using analytical LC-MS.

Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Example 84. Analysis results for the 1st sample injection: purity: 100.0%; retention time: 2.53; observed adducts: [M+H]; measured masses: 581.96. Analysis results for the 2nd sample injection: purity: 100.0%; retention time: 1.84; observed adducts: [M+H]; measured masses: 581.97. Example 84 (5.1 mg) was isolated in 5.6% yield.

Example 85. Analysis results for the 1st sample injection: purity: 100.0%; retention time: 2.53; observed adducts: [M+H]; measured masses: 581.95. Analysis results for the 2nd sample injection: purity: 99.0%; retention time: 1.83; observed adducts: [M+H]; measured masses: 581.97. Example 85 (5.3 mg) was isolated in 5.8% yield.

Example 86 1 N)-on

- 101 044500

2-(( 4-fluorophenyl)(piperazin-1-yl)methyl)-4-methylphenol (70.7 mg, 0.235 mmol) and then DIPEA (0.082 ml, 0.471 mmol) was added. This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-90% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 70°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 2.0 min; flow rate: 0.75 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters CSH C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 70°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 2.0 min; flow rate: 0.75 ml/min; registration: UV, 220 nm. Purity: 100.0%; retention time: 2.5; observed adducts: [M+H]; measured masses: 582.08. Analysis results for the 2nd sample injection: purity: 97.4%; retention time: 1.82; observed adducts: [M+H]; measured masses: 581.86. The title compound (5.3 mg) was isolated in 5.8% yield.

Example 87 1 N)-on

2-((4- fluorophenyl)(piperazin-1-yl)methyl)-3-methylphenol (141 mg, 0.471 mmol) and then DIPEA (0.165 ml, 0.942 mmol) was added. This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was diluted with ethyl acetate, washed with water and brine, and dried over magnesium sulfate. The residue was purified on a BIOTAGE™ MPLC system using hexanes:ethyl acetate (1:1) and a silica gel column (40 g). Fractions containing the desired product were combined to give the product as a yellow substance. The title compound (100 mg) was isolated in 54.7% yield. Approximately one third of the resulting yellow solid was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; gradient: 30-70% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature:

- 102 044500

50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 1st sample input: purity: 100.0%; retention time: 2.4; observed adducts: [M+H]; measured masses: 582. Analysis results for the 2nd sample injection: purity: 97.2%; retention time: 1.7; observed adducts: [M+H]; measured masses: 581.94. 33.4 mg of the title compound was isolated.

Examples 88-90. 5-((2-((4-(6-Bromo-1-methyl-3-nitro-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)piperazin-1-yl)( 4-fluorophenyl)methyl)-3-methylphenoxy)methyl)nicotinonitrile

To a solution of 6-bromo-4-(4-((4-fluorophenyl)(2-hydroxy-6-methylphenyl)methyl)piperazin-1 -yl)-1 -methyl3-nitro-1,5-naphthyridin-2(1H )-one (30 mg, 0.052 mmol) in DMF (2 ml) was added cesium carbonate (50.3 mg, 0.155 mmol). This reaction mixture was stirred for 20 minutes at room temperature. 5-(chloromethyl)nicotinonitrile (23.58 mg, 0.155 mmol) was added and the reaction mixture was heated for 3 hours at 75°C. LC-MS analysis indicated approximately 40% conversion of the starting materials. The reaction mixture was stirred by shaking at 75°C overnight. LC-MS analysis indicated approximately 75% conversion of the starting materials. The reaction mixture was stirred by shaking at 75°C for an additional 24 hours. According to LC-MS analysis, the reaction was almost complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: Waters CSH C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; gradient: 18-58% B for 28 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. LC-MS results (acetonitrile TFA)-3: purity: 98.7%; retention time: 1.72; observed adducts: [M+H]; measured masses: 697.92; LC-MS results (acetonitrile ammonium acetate)-3: purity: 96.6%; retention time: 2.49; observed adducts: [M+H]; measured masses: 697.9. The title racemic compound (21.7 mg) was isolated in 59.7% yield. The racemic substance was further purified using SFC-chiral chromatography to obtain isomers.

The compound of Example 89 (first eluting isomer) and the compound of Example 90 (second eluting isomer) were isolated from the racemate using chiral SFC chromatography. To determine the final purity of the products, samples were analyzed in duplicate using analytical LC-MS.

Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min;

- 103 044500 registration: UV, 220 nm.

Example 89. Analysis results for the 1st sample injection: purity: 98.3%; retention time: 2.51;

observed adducts: [M+H]; measured masses: 697.93. Analysis results for 2nd sample injection:

purity: 99.0%; retention time: 1.68; observed adducts: [M+H]; measured masses: 697.93. Example 89 (5.4 mg) was isolated in 14.9% yield.

Example 90. Analysis results for the 1st sample injection: purity: 99.2%; retention time: 2.51; observed adducts: [M+H]; measured masses: 697.9. Analysis results for the 2nd sample injection: purity: 100.0%; retention time: 1.69; observed adducts: [M+H]; measured masses: 697.92. Example 90 (5.9 mg) was isolated in 16.2% yield.

Examples 91-93. 5-((2-((4-(6-Bromo-1 -methyl-3-nitro-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)piperazin-1-yl)( 4-fluorophenyl)methyl)-5-fluorophenoxy)methyl)nicotinonitrile

To a solution of 6-bromo-4-(4-((4-fluoro-2-hydroxyphenyl)(4-fluorophenyl)methyl)piperazin-1-yl)-1-methyl-3nitro-1,5-naphthyridin-2(1H )-one (20 mg, 0.034 mmol) in DMF (1.6 ml) was added cesium carbonate (33.3 mg, 0.102 mmol). This reaction mixture was stirred for 20 minutes at room temperature. 5-(chloromethyl)nicotinonitrile (15.61 mg, 0.102 mmol) was added and the reaction mixture was heated at 75 °C overnight. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: Waters CSH C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; gradient: 15-55% B for 25 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. LC-MS results (acetonitrile TFA)-2: purity: 99.0%; retention time: 1.74; observed adducts: [M+H]; measured masses: 701.88; LC-MS results (acetonitrile ammonium acetate)-2: purity: 98.8%; retention time: 2.4; observed adducts: [M+H]; measured masses: 701.91. The title racemic compound (5.2 mg) was isolated in 21.8% yield. The racemic substance was further purified using SFC-chiral chromatography to obtain isomers.

The compound of Example 92 (first eluting isomer) and the compound of Example 93 (second eluting isomer) were isolated from the racemate using SFC-chiral chromatography. To determine the final purity of the products, samples were analyzed in duplicate using analytical LC-MS.

Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Example 92. Analysis results for the 1st sample injection: purity: 100.0%; retention time: 2.4; observed adducts: [M+H]; measured masses: 701.91. Analysis results for 2nd sample injection:

- 104 044500 purity: 100.0%; retention time: 1.7; observed adducts: [M+H]; measured masses: 701.81. Example 92 (1.4 mg) was isolated in 5.9% yield.

Example 93. Analysis results for the 1st sample injection: purity: 100.0%; retention time: 2.4; observed adducts: [M+H]; measured masses: 701.92. Analysis results for the 2nd sample injection: purity: 100.0%; retention time: 1.7; observed adducts: [M+H]; measured masses: 701.94. Example 93 (1.5 mg) was isolated in 6.3% yield.

Examples 94-96. 4-(4-((2-(Allyloxy)-6-methylphenyl)(4-fluorophenyl)methyl)piperazin-1-yl)-6-bromo1 -methyl-3-nitro-1,5-naphthyridine-2(1H )-He

CH ₃

To a solution of 6-bromo-4-(4-((4-fluorophenyl)(2-hydroxy-6-methylphenyl)methyl)piperazin-1 -yl)-1 -methyl3-nitro-1,5-naphthyridin-2(1H )-one (20 mg, 0.034 mmol) in DMF (1.6 ml) was added cesium carbonate (33.6 mg, 0.103 mmol). This reaction mixture was stirred for 20 minutes at room temperature. Allyl bromide (8.91 μl, 0.103 mmol) was added and the reaction mixture was stirred overnight at 60°C. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-100% B for 15 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. The resulting substance was further purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; gradient: 7-47% B for 25 min, then maintain 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Purity: 100.0%; retention time: 1.59; observed adducts: [M+H]; measured masses: 621.9. Analysis results for the 1st sample input: purity: 100.0%; retention time: 2.4; observed adducts: [M+H]; measured masses: 621.92. The title racemic compound (16.9 mg) was isolated in 79.9% yield. The racemic substance was further purified using SFC-chiral chromatography to obtain isomers.

The compound of Example 95 (first eluting isomer) and the compound of Example 96 (second eluting isomer) were isolated from the racemate using SFC-chiral chromatography. To determine the final purity of the products, samples were analyzed in duplicate using analytical LC-MS.

Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Example 95. Analysis results for the 1st sample injection: purity: 97.8%; retention time: 2.77;

- 105 044500 observed adducts: [M+H]; measured masses: 621.94. Analysis results for 2nd sample injection:

purity: 100.0%; retention time: 1.83; observed adducts: [M+H]; measured masses: 621.96.

Example 95 (5.3 mg) was isolated in 25% yield.

Example 96. Analysis results for the 1st sample injection: purity: 98.4%; retention time: 2.77; observed adducts: [M+H]; measured masses: 621.94. Analysis results for the 2nd sample injection: purity: 100.0%; retention time: 1.83; observed adducts: [M+H]; measured masses: 621.93. Example 96 (5.3 mg) was isolated in 25% yield.

Examples 97-99. 8-(4-((4-Fluorophenyl)(2-hydroxyphenyl)methyl)piperazin-1-yl)-5-methyl-7-nitro-6oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile

CH ₃

2-( (4-fluorophenyl)(piperazin-1-yl)methyl)phenol (390 mg, 1.360 mmol) and then potassium carbonate (313 mg, 2.267 mmol) was added. This reaction mixture was stirred overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was diluted with ethyl acetate, washed with brine and dried over magnesium sulfate. The residue was purified on a BIOTAGE™ MPLC system using a gradient of (1:1) hexanes:ethyl acetate to 100% ethyl acetate and a silica gel column (24 g). After collecting the fractions, the desired product was obtained as a light yellow solid. This substance was further purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 43-83% B for 15 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LCMS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 1st sample input: purity: 100.0%; retention time: 2.17; observed adducts: [M+H]; measured masses: 514.99. Analysis results for the 2nd sample injection: purity: 98.1%; retention time: 1.51; observed adducts: [M+H]; measured masses: 515.01. The title racemic compound (220 mg) was isolated in 37.7% yield. The racemic substance was further purified using SFC-chiral chromatography to obtain isomers.

A portion of the racemic substance (30 mg) was further purified using SFC-chiral chromatography to obtain the compound of Example 98 (first eluting isomer) and the compound of Example 99 (second eluting isomer). To determine the final purity of the products, samples were analyzed in duplicate using analytical LC-MS.

Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 2nd sample injection: purity: 100.0%; retention time: 1.55; observed adducts: [M+H]; measured masses: 515.03.

- 106 044500

Example 98. Analysis results for the 1st sample injection: purity: 100.0%; retention time: 2.2;

observed adducts: [M+H]; measured masses: 515.03. Example 98 (3.3 mg) was isolated in 11% yield.

Example 99. Analysis results for the 1st sample injection: purity: 100.0%; retention time: 2.17; observed adducts: [M+H]; measured masses: 515.01. Analysis results for the 2nd sample injection: purity: 100.0%; retention time: 1.5; observed adducts: [M+H]; measured masses: 515.01. Example 99 (4.2 mg) was isolated in 14% yield.

Example 100 ,6-dihydro-1,5-naphthyridine-2-carbonitrile

To a solution of 8-(4-((4-fluorophenyl)(2-hydroxyphenyl)methyl)piperazin-1-yl)-5-methyl-7-nitro-6-oxo5,6-dihydro-1,5-naphthyridin-2 -carbonitrile (40 mg, 0.078 mmol) in DMF (1 ml), potassium carbonate (32.2 mg, 0.233 mmol) was added and then propargyl bromide (80% (w/w) in toluene) (0.026 ml, 0.233 mmol) was added. This reaction mixture was stirred by shaking at 75°C over the weekend. According to LC-MS analysis, approximately 40% of the starting material was converted to the desired product. Additional propargyl bromide (80% (w/w) in toluene) (0.026 mL, 0.233 mmol) was added at the same time as cesium carbonate (38.0 mg, 0.117 mmol). The reaction mixture was stirred by shaking at 75°C overnight. According to LC-MS analysis, the starting material was completely consumed. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 40-85% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 553.08; retention time: 1.61 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 553.07; retention time: 2.41 min. The title racemic compound (13.5 mg) was isolated in 31.3% yield. The racemic substance was further purified using SFC-chiral chromatography to obtain isomers.

Example 101 8-(4-((4-Fluorophenyl)(2-(prop-2-yn-1-yloxy)phenyl)methyl)piperazin-1-yl)-5-methyl-7nitro-6-oxo-5 ,6-dihydro-1,5-naphthyridine-2-carbonitrile (first eluting isomer)

The resulting racemic substance was further purified using SFC chiral chromatography. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B:

- 107 044500 (95:5) acetonitrile: water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 553.08; retention time: 1.66 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 553.12; retention time: 2.33 min. The title compound (4 mg) was isolated in 9.3% yield.

Example 102 8-(4-((4-Fluorophenyl)(2-(prop-2-yn-1-yloxy)phenyl)methyl)piperazin-1-yl)-5-methyl-7nitro-6-oxo-5 ,6-dihydro-1,5-naphthyridine-2-carbonitrile (second eluting isomer)

The resulting racemic substance was further purified using SFC chiral chromatography. The crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 40-85% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 553.07; retention time: 2.33 min. The title compound (3.7 mg) was isolated in 8.6% yield.

Example 103 4-(4-(bis(4-Fluorophenyl)methyl)piperazin-1-yl)-6-methoxy-1-methyl-2-oxo-1,2-dihydro1,5-naphthyridine-3-carbonitrile

Huenig's base ( 0.014 ml, 0.080 mmol) and then 1(bis(4-fluorophenyl)methyl)piperazine (23.10 mg, 0.080 mmol) was added. This reaction mixture was stirred by shaking for 2 hours at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 45-85% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; poto speed

- 108 044500 ka: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 99.0%; measured mass: 502.09; retention time: 1.5 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 502.09; retention time: 2.25 min. The title compound (3.4 mg) was isolated in 8.5% yield.

Example 104 3-carbonitrile

Huenig's base ( 0.014 ml, 0.080 mmol) and then 2((4-fluorophenyl)(piperazin-1-yl)methyl) phenol, 2 TFA (41.2 mg, 0.080 mmol) was added. This reaction mixture was stirred by shaking for 2 hours at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; gradient: 17-57% B for 15 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 500.09; retention time: 2.1 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 500.09; retention time: 1.4 min. The title compound (2.2 mg) was isolated in 5.5% yield.

Examples 105-107. 6-Bromo-4-(4-((4-fluorophenyl)(2-hydroxyphenyl)methyl)piperazin-1-yl)-2-oxo-1(prop-2-yn-1 -yl)-1,2- Dihydro-1,5-naphthyridine-3-carbonitrile

Solution of 2-hydroxybenzaldehyde (15.07 mg, 0.123 mmol), 6-bromo-2-oxo-4-(piperazin-1-yl)-1(prop-2-yn-1-yl)-1,2-dihydro -1,5-naphthyridine-3-carbonitrile TFA (60 mg, 0.123 mmol) and (4-fluorophenyl)boronic acid (17.27 mg, 0.123 mmol) in DMF (1 ml) were heated in a microwave oven at 150 °C in for 2 hours. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-90% B in those

- 109 044500 reading for 18 minutes, then maintaining 100% B for 5 minutes; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 97.3%; measured mass: 572.05; retention time: 1.85 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 97.4%; measured mass: 572.1; retention time: 2.75 min. The title racemic compound (15.3 mg) was isolated in 21.7% yield. The racemic substance was further purified using SFC chiral chromatography to obtain isomers.

The racemic compound of Example 105 was further purified using SFC-chiral chromatography to obtain the compound of Example 106 (first eluting isomer) and the compound of Example 107 (second eluting isomer). Analytical LC-MS was used to determine the final purity of the products.

Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Example 106. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 572.1; retention time: 1.91 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 572.08; retention time: 2.72 min. Example 106 (6.8 mg) was isolated in 9.7% yield.

Example 107. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 572.06; retention time: 1.91 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 572.09; retention time: 2.72 min. Example 107 (7.4 mg) was isolated in 10.5% yield.

Example 113 3-carbonitrile

To a solution of triethylamine (0.017 ml, 0.123 mmol), 1-methylpiperidine (0.015 ml, 0.123 mmol) and 6bromo-4-hydroxy-1-(2-methoxyethyl)-2-oxo-1,2-dihydro-1,5- naphthyridine-3-carbonitrile (20 mg, 0.062 mmol) in acetonitrile (1 ml) was added 4-methylbenzene-1-sulfonyl chloride (23.53 mg, 0.123 mmol). This reaction mixture was stirred by shaking for 30 minutes at room temperature. 1-(bis(4-fluorophenyl)methyl)piperazine (35.6 mg, 0.123 mmol) was then added. The reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, all the starting material was consumed. The desired product was detected as a small peak and purified using reverse phase HPLC for purification. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-100% B for 20 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing

- 110 044500 the desired product was combined and dried using a centrifugal evaporator. The resulting substance was further purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; gradient: 25-65% B for 20 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 594.09; retention time: 1.78 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 594.1; retention time: 2.56 min. The title compound (1.8 mg) was isolated in 4.9% yield.

Examples 114-116. 6-Bromo-4-(4-((4-fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl)piperazin-1-yl)-1(2-methoxyethyl)-2-oxo-1,2-dihydro- 1,5-naphthyridine-3-carbonitrile

To a solution of 6-bromo-4-chloro-1-(2-methoxyethyl)-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (20 mg, 0.058 mmol) in DMF (1 ml) Hunig's base (0.051 mL, 0.292 mmol) and 1-((4-fluoro2-methoxyphenyl)(4-fluorophenyl)methyl)piperazine, 2 TFA (31.9 mg, 0.058 mmol) were added. This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude substance was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 60-100% B for 19 min, then maintain 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 im). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 624.06; retention time: 1.76 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 624.07; retention time: 2.59 min. The racemic compound of Example 114 (19.7 mg) was isolated in 54.4% yield.

The compound of Example 114 was further purified using SFC-chiral chromatography to obtain the compound of Example 115 (first eluting isomer) and the compound of Example 116 (second eluting isomer). Analytical LC-MS was used to determine the final purity of the products.

Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; re

- 111 044500 hystration: MS and UV (220 nm).

Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Example 115. Analysis results for the 1st sample injection: purity: 100.0%; measured weight: 624.05; retention time: 1.76 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 624.06; retention time: 2.58 min. Example 115 (4.5 mg) was isolated in 12.4% yield.

Example 116. Analysis results for the 1st sample injection: purity: 100.0%; measured weight: 624.05; retention time: 1.76 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 624.07; retention time: 2.58 min. Example 116 (4.2 mg) was isolated in 11.6% yield.

Example 117 1,5-naphthyridine-3-carbonitrile

To a solution of 6-bromo-4-chloro-1-(2-methoxyethyl)-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (20 mg, 0.058 mmol) in DMF (1 ml) Hunig's base (0.041 mL, 0.234 mmol) and 2-((4fluorophenyl)(piperazin-1-yl)methyl)phenol (16.72 mg, 0.058 mmol) were added. This reaction mixture was stirred by shaking for 1 hour at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-90% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 592.04; retention time: 2.29 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 592.03; retention time: 1.54 min. The title compound (6.4 mg) was isolated in 18.6% yield.

Example 118 1,5-naphthyridine-2-carbonitrile

To a solution of 8-chloro-7-nitro-6-oxo-5-(prop-2-in-1-yl)-5,6-dihydro-1,5-naphthyridine-2-carbonitrile

- 112 044500 (25 mg, 0.087 mmol) in DMF (1.5 ml) was added 1-(bis(4-fluorophenyl)methyl)piperazine (24.97 mg, 0.087 mmol) and then Huenig's base (0.08 ml) was added , 0.43 mmol). This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-90% B for 22 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 541.08; retention time: 1.74 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 541.09; retention time: 2.38 min. The title compound (32.4 mg) was isolated in 68.9% yield.

Examples 119-121. 8-(4-((4-Fluoro-2-hydroxyphenyl)(4-fluorophenyl)methyl)piperazin-1-yl)-7-nitro-6oxo-5-(prop-2-in-1-yl)-5 ,6-dihydro-1,5-naphthyridine-2-carbonitrile

To a solution of 8-chloro-7-nitro-6-oxo-5-(prop-2-yn-1-yl)-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (25 mg, 0.087 mmol) in DMF (1.5 ml) added Huenig's base (0.076 ml, 0.433 mmol) and then added 5-fluoro-2-((4-fluorophenyl)(piperazin-1-yl)methyl)phenol, TFA (36.2 mg , 0.087 mmol). This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, 2 main peaks appeared. The resulting crude substance was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-100% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 557.11; retention time: 2.31 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 557.11; retention time: 1.69 min. The title compound (10.5 mg) was isolated in 21.7% yield.

A portion of the racemic substance (8.37 mg) was further purified using SFC-chiral chromatography to obtain the compound of Example 120 (first eluting isomer) and the compound of Example 121 (second eluting isomer). Analytical LC-MS was used to determine the final purity of the products.

Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size

- 113 044500

1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase

B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to

100% B for 3 min, then maintain 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Example 120. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 557.13; retention time: 1.7 min. Analysis results for the 2nd sample injection: purity: 98.6%; measured mass: 557.17; retention time: 2.32 min. Example 120 (3.5 mg) was isolated in 41.9% yield.

Example 121. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 557.16; retention time: 1.7 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 557.12; retention time: 2.32 min. Example 121 (3.2 mg) was isolated in 38.3% yield.

Example 122 1,5-naphthyridine-2-carbonitrile

To a solution of 8-chloro-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (15 mg, 0.057 mmol) in DMF (1.5 ml) was added carbonate potassium (23.50 mg, 0.170 mmol) and then 5fluoro-2-((4-fluorophenyl)(piperazin-1-yl)methyl)phenol, TFA (23.71 mg, 0.057 mmol) was added. This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, 2 main peaks appeared. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; gradient: 10-80% B for 27 min, then maintain 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 98.2%; measured mass: 533.11; retention time: 1.53 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 97.8%; measured mass: 533.09; retention time: 2.17 min. The title compound (3.4 mg) was isolated in 11.2% yield.

Examples 123-125. 8-(4-((4-Fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl)piperazin-1-yl)-5-methyl-7nitro-6-oxo-5,6-dihydro-1,5-naphthyridine -2-carbonitrile

- 114 044500

Huenig's base ( 0.099 mL, 0.567 mmol) and then 1((4-fluoro-2-methoxyphenyl)(4-fluorophenyl) methyl)piperazine, 2 TFA (61.9 mg, 0.113 mmol) was added. This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 48-88% B for 19 min, then maintain 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 547.16; retention time: 2.41 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 547.15; retention time: 1.67 min. The title racemic compound (18 mg) was isolated in 29.1% yield.

The racemic material was further purified using SFC-chiral chromatography to obtain the compound of Example 124 (first eluting isomer) and the compound of Example 125 (second eluting isomer). Analytical LCMS was used to determine the final purity of the products.

Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 547.14; retention time: 2.41 min.

Example 124. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 547.15; retention time: 1.66 min. Analysis results for the 2nd sample injection: purity: 100.0%; retention time: 1.66; observed adducts: [M+H]; measured masses: 547.15. Example 124 (8.1 mg) was isolated in 13.1% yield.

Example 125. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 547.21; retention time: 1.66 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 547.14; retention time: 2.41 min. Example 125 (8.1 mg) was isolated in 13.1% yield.

Examples 126-128. 8-(4-((4-Fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl)piperazin-1-yl)-7-nitro-6oxo-5-(prop-2-in-1-yl)-5 ,6-dihydro-1,5-naphthyridine-2-carbonitrile

To a solution of 8-chloro-7-nitro-6-oxo-5-(prop-2-yn-1-yl)-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (25 mg, 0.087 mmol) 1-((4-fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl)piperazine, 2 TFA (47.3 mg, 0.087 mmol) were added to DMF (1.5 ml) and then Huenig's base (0.08 ml, 0.43 mmol). This reaction mixture was stirred by shaking overnight at room temperature. Co

- 115 044500 According to LC-MS analysis, the reaction was completed. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-100% B for 20 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 571.14; retention time: 2.47 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 571.17; retention time: 1.73 min. The title racemic compound (21.3 mg) was isolated in 42.9% yield.

The racemic material was further purified using SFC-chiral chromatography to obtain the compound of Example 127 (first eluting isomer) and the compound of Example 128 (second eluting isomer). Analytical LCMS was used to determine the final purity of the products.

Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Example 127. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 571.12; retention time: 1.75 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 571.16; retention time: 2.48 min. Example 127 (7.7 mg) was isolated in 15.5% yield.

Example 128. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 571.15; retention time: 1.74 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 571.16; retention time: 2.48 min. Example 128 (7.5 mg) was isolated in 15.1% yield.

Examples 129-131. 8-(4-((4-Fluorophenyl)(2-methoxyphenyl)methyl)piperazin-1-yl)-7-nitro-6-oxo-5(prop-2-yn-1-yl)-5,6- Dihydro-1,5-naphthyridine-2-carbonitrile

To a solution of 8-chloro-7-nitro-6-oxo-5-(prop-2-yn-1-yl)-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (25 mg, 0.087 mmol) in DMF (1.5 ml) was added 1-((4-fluorophenyl)(2-methoxyphenyl)methyl)piperazine, 2 TFA (45.8 mg, 0.087 mmol) and then Huenig's base (0.08 ml, 0.087 mmol) was added 43 mmol). This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-95% B for 20 min, then maintenance

- 116 044500

100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 553.15; retention time: 1.67 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 553.15; retention time: 2.45 min. The title racemic compound (27.3 mg) was isolated in 56.8% yield.

The racemic material was further purified using SFC-chiral chromatography to obtain the compound of Example 130 (first eluting isomer) and the compound of Example 131 (second eluting isomer). Analytical LCMS was used to determine the final purity of the products.

Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Example 130. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 553.15; retention time: 2.45 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 553.13; retention time: 1.68 min. Example 130 (8 mg) was isolated in 16.6% yield.

Example 131. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 553.14; retention time: 2.45 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 553.17; retention time: 1.71 min. Example 131 (2.3 mg) was isolated in 4.8% yield.

Examples 132-134. 8-(4-((4-Fluorophenyl)(2-hydroxyphenyl)methyl)piperazin-1-yl)-7-nitro-6-oxo-5(prop-2-yn-1-yl)-5,6- Dihydro-1,5-naphthyridine-2-carbonitrile

To a solution of 8-chloro-7-nitro-6-oxo-5-(prop-2-yn-1-yl)-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (25 mg, 0.087 mmol) in DMF (1.5 ml) was added 2-((4-fluorophenyl)(piperazin-1-yl)methyl)phenol, 2 TFA (44.6 mg, 0.087 mmol) and then Huenig's base (0.08 ml, 0.43 mmol). This reaction mixture was stirred by shaking for 1 hour at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-90% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for 1st sample injection: purity:

- 117 044500

100.0%; measured mass: 539.13; retention time: 2.26 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 539.14; retention time: 1.62 min. The title racemic compound (9.7 mg) was isolated in 20.7% yield.

The racemic material was further purified using SFC-chiral chromatography to obtain the compound of Example 133 (first eluting isomer) and the compound of Example 134 (second eluting isomer). Analytical LCMS was used to determine the final purity of the products.

Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Example 133. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 539.17; retention time: 2.25 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 539.15; retention time: 1.61 min. Example 133 (4.3 mg) was isolated in 9.2% yield.

Example 134. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 539.12; retention time: 2.25 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 539.19; retention time: 1.6 min. Example 134 (2.8 mg) was isolated in 6% yield.

Examples 135-137. 8-(4-( 1 -(4-Fluorophenyl)ethyl)piperazin-1 -yl)-7-nitro-6-oxo-5-(prop-2-in-1 -yl)5,6-dihydro-1 ,5-naphthyridine-2-carbonitrile

To a solution of 8-chloro-7-nitro-6-oxo-5-(prop-2-yn-1-yl)-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (20 mg, 0.055 mmol) and 1-(1-phenylethyl)piperazine (46.4 mg, 0.111 mmol) in DMF (1 ml) added Huenig's base (0.029 ml, 0.166 mmol) and 1-(1-phenylethyl)piperazine (46.4 mg, 0.111 mmol). This reaction mixture was stirred by shaking for 2 hours at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; gradient: 13-53% B for 20 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 461.16; retention time: 1.33 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass:

- 118 044500

461.11; retention time: 2.15 min. The title racemic compound (20.5 mg) was isolated in 80.9% yield.

The racemic material was further purified using SFC-chiral chromatography to obtain the compound of Example 136 (first eluting isomer) and the compound of Example 137 (second eluting isomer). Analytical LCMS was used to determine the final purity of the products.

Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Example 136. Analysis results for the 1st sample injection: purity: 99.0%; measured mass: 461.12; retention time: 1.35 min. Analysis results for the 2nd sample injection: purity: 98.8%; measured mass: 461.12; retention time: 2.15 min. Example 136 (6.1 mg) was isolated in 24.1% yield.

Example 137. Analysis results for the 1st sample injection: purity: 98.6%; measured mass: 461.14; retention time: 1.35 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 461.13; retention time: 2.15 min. Example 137 (5.7 mg) was isolated in 22.5% yield.

Example 138 1-yl)-5,6-dihydro-1,5-naphthyridine-2-carbonitrile

To a solution of 8-chloro-7-nitro-6-oxo-5-(prop-2-yn-1-yl)-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (20 mg, 0.055 mmol) and Huenig's base (0.029 ml, 0.166 mmol) in DMF (1 ml) was added 1-(6methoxy-2,3-dihydro-1H-inden-1-yl)piperazine (51.0 mg, 0.111 mmol). This reaction mixture was stirred by shaking for 2 hours at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude substance was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; gradient: 15-55% B for 20 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 485.18; retention time: 2.13 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 485.16; retention time: 1.4 min. The title compound (24 mg) was isolated in 90.1% yield.

Examples 139-141. 8-{4-[1 -(4-Fluorophenyl)propyl]piperazin-1 -yl}-7-nitro-6-oxo-5-(prop-2-in-1 yl)-5,6-dihydro-1 ,5-naphthyridine-2-carbonitrile

- 119 044500

To a solution of 8-chloro-7-nitro-6-oxo-5-(prop-2-yn-1-yl)-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (20 mg, 0.055 mmol) and 1-(1-phenylpropyl)piperazine, 2 TFA (35.9 mg, 0.083 mmol) in DMF (1 ml) was added Huenig's base (0.048 ml, 0.277 mmol) and 1-(1-phenylpropyl)piperazine, 2 TFA ( 35.9 mg, 0.083 mmol). This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 45-85% B for 20 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 475.16; retention time: 1.45 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 475.16; retention time: 2.29 min. The title racemic compound (15.3 mg) was isolated in 58.6% yield.

The racemic material was further purified using SFC-chiral chromatography to obtain the compound of Example 140 (first eluting isomer) and the compound of Example 141 (second eluting isomer). Analytical LCMS was used to determine the final purity of the products. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Example 140. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 475.19; retention time: 2.28 min. Analysis results for the 2nd sample injection: purity: 97.3%; measured mass: 475.19; retention time: 1.47 min. Example 140 (6.9 mg) was isolated in 26.4% yield.

Example 141. Analysis results for the 1st sample injection: purity: 98.2%; measured mass: 475.17; retention time: 2.28 min. Analysis results for the 2nd sample injection: purity: 98.5%; measured mass: 475.18; retention time: 1.47 min. Example 141 (6.4 mg) was isolated in 24.5% yield.

Examples 142-144. 8-(4-(Cyclopropyl(4-fluorophenyl)methyl)piperazin-1-yl)-7-nitro-6-oxo-5-(prop2-in-1-yl)-5,6-dihydro-1,5 -naphthyridine-2-carbonitrile

- 120 044500

To a solution of 1-(cyclopropyl(4-fluorophenyl)methyl)piperazine, TFA (24.14 mg, 0.069 mmol) and 8-chloro7-nitro-6-oxo-5-(prop-2-in-1-yl)- 5,6-dihydro-1,5-naphthyridine-2-carbonitrile (20 mg, 0.069 mmol) in DMF (1 ml) was added Huenig's base (0.036 ml, 0.208 mmol). This reaction mixture was stirred by shaking for 2 hours at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 45-85% B for 20 min, then maintaining 100% B for 6 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator.

The resulting racemic material was purified using SFC-chiral chromatography to yield the compound of Example 143 (first eluting isomer) and the compound of Example 144 (second eluting isomer). Analytical LC-MS was used to determine the final purity of the products.

Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Example 143. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 487.02; retention time: 1.56 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 487.02; retention time: 2.23 min. Example 143 (3.8 mg) was isolated in 11.3% yield.

Example 144. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 487.03; retention time: 1.41 min. Analysis results for the 2nd sample injection: purity: 99.1%; measured mass: 487.03; retention time: 2.23 min. Example 144 (4 mg) was isolated in 11.9% yield.

Example 145 5-naphthyridine-2-carbonitrile

To a solution of 8-chloro-5-(2-methoxyethyl)-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (5 mg, 0.016 mmol) in DMF (0.5 ml) Hunig's base (8.49 µl, 0.049 mmol) and 2-((4fluorophenyl)(piperazin-1-yl)methyl)phenol (9.28 mg, 0.032 mmol) were added. This reaction mixture was stirred by shaking for 1 hour at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 38-78% B for 20 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using

- 121 044500 called a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 559.23; retention time: 1.6 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 559.19; retention time: 2.27 min. The title compound (1.5 mg) was isolated in 16.8% yield.

Example 146 6-dihydro-1,5-naphthyridine-2-carbonitrile

To a solution of 8-chloro-5-(2-methoxyethyl)-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (15 mg, 0.049 mmol) in DMF (0.5 ml) Hunig's base (0.025 ml, 0.146 mmol) and 1-(6methoxy-2,3-dihydro-1H-inden-1-yl)piperazine (16.93 mg, 0.073 mmol) were added. This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 40-80% B for 20 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 505.21; retention time: 1.41 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 505.2; retention time: 2.13 min. The title compound (12.5 mg) was isolated in 50.6% yield.

Example 147 2-carbonitrile

To a solution of 8-chloro-5-(2-methoxyethyl)-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (15 mg, 0.049 mmol) in DMF (0.5 ml) Hunig's base (0.025 ml, 0.146 mmol) and 1-(bis(4

- 122 044500 fluorophenyl)methyl)piperazine (21.02 mg, 0.073 mmol). This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 46-86% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 561.17; retention time: 1.79 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 561.17; retention time: 2.46 min. The title compound (13.5 mg) was isolated in 49.1% yield.

Examples 148-150. 8-(4-(1-(4-Fluorophenyl)ethyl)piperazin-1-yl)-5-(2-methoxyethyl)-7-nitro-6-oxo5,6-dihydro-1,5-naphthyridin-2- carbonitrile

To a solution of 8-chloro-5-(2-methoxyethyl)-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (15 mg, 0.049 mmol) in DMF (0.5 ml) Hunig's base (0.051 ml, 0.292 mmol) and 1-(1-(4fluorophenyl)ethyl)piperazine dihydrochloride (aq) (20.50 mg, 0.073 mmol) were added. This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 38-78% B for 22 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 481.19; retention time: 1.36 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 481.17; retention time: 2.16 min. The title racemic compound (11.5 mg) was isolated in 48.8% yield.

The racemic material was further purified using SFC-chiral chromatography to obtain the compound of Example 149 (first eluting isomer) and the compound of Example 150 (second eluting isomer). Analytical LCMS was used to determine the final purity of the products.

Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0% V to 100% V

- 123 044500 for 3 minutes, then maintaining 100% B for 0.75 minutes; flow rate: 1 ml/min; registration:

MS and UV (220 nm).

Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Example 149. Analysis results for the 1st sample injection: purity: 97.1%; measured mass: 481.18; retention time: 2.17 min. Analysis results for the 2nd sample injection: purity: 97.4%; measured mass: 481.16; retention time: 1.36 min. Example 149 (3.7 mg) was isolated in 15.7% yield.

Example 150. Analysis results for the 1st sample injection: purity: 97.1%; measured mass: 481.2; retention time: 2.17 min. Analysis results for the 2nd sample injection: purity: 97.3%; measured mass: 481.18; retention time: 1.36 min. Example 150 (3.6 mg) was isolated in 15.3% yield.

Example 151 5-(Cyanomethyl)-8-(4-(1-(4-fluorophenyl)ethyl)piperazin-1-yl)-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine -2-carbonitrile

To a solution of 8-chloro-5-(cyanomethyl)-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (15 mg, 0.052 mmol) in DMF (0.5 ml) Hunig's base (0.027 mL, 0.155 mmol) and 1-(1-(4fluorophenyl)ethyl)piperazine (16.18 mg, 0.078 mmol) were added. This reaction mixture was stirred by shaking for 1 hour at room temperature. According to HPLC analysis, the reaction was approximately 80% complete. The reaction mixture was stirred by shaking at room temperature for more than 3 hours. The resulting crude substance was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 28-68% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 462.14; retention time: 1.28 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 462.15; retention time: 2.07 min. The title compound (6.6 mg) was isolated in 27.5% yield.

Example 152 2-carbonitrile

- 124 044500

To a solution of 8-chloro-5-(cyanomethyl)-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (15 mg, 0.052 mmol) in DMF (0.5 ml) Hunig's base (0.027 mL, 0.155 mmol) and 1-(bis(4fluorophenyl)methyl)piperazine (22.40 mg, 0.078 mmol) were added. This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 41-81% B for 23 min, then maintain 100% B for 6 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 542.09; retention time: 1.71 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 542.11; retention time: 2.32 min. The title compound (5.3 mg) was isolated in 18.8% yield.

Example 153 3-carbonitrile

To a solution of 6-bromo-4-chloro-1-(cyanomethyl)-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (15 mg, 0.046 mmol) in DMF (0.5 ml) Hunig's base (0.024 mL, 0.139 mmol) and 1-(bis(4fluorophenyl)methyl)piperazine (20.05 mg, 0.070 mmol) were added. This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 43-83% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 97.1%; measured mass: 577.13; retention time: 2.39 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 97.7%; measured mass: 575.07; retention time: 1.74 min. The title compound (6.3 mg) was isolated in 23.8% yield.

Example 154 -3-carbonitrile

- 125 044500

To a solution of 6-bromo-4-chloro-1-(cyanomethyl)-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (15 mg, 0.046 mmol) in DMF (0.5 ml) Hunig's base (0.024 mL, 0.139 mmol) and 1-(1-(4fluorophenyl)ethyl)piperazine (14.48 mg, 0.070 mmol) were added. This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 38-78% B for 20 min, then maintaining 100% B for 6 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 494.97; retention time: 2.05 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 494.95; retention time: 1.26 min. The title compound (6.6 mg) was isolated in 29% yield.

Example 155 -3-carbonitrile

To a solution of 6-bromo-4-chloro-1-(cyanomethyl)-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (15 mg, 0.046 mmol) in DMF (0.5 ml) Hunig's base (0.024 mL, 0.139 mmol) and 1-(1-(4fluorophenyl)propyl)piperazine (15.46 mg, 0.070 mmol) were added. This reaction mixture was stirred by shaking overnight at room temperature. According to HPLC analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 38-78% B for 25 min, then maintaining 100% B for 6 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 509.07; retention time: 1.43 min. Analysis conditions for the 2nd sample injection: column 126 044500: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 509.1; retention time: 2.26 min. The title compound (6.8 mg) was isolated in 29% yield.

Example 156 6-Bromo-1-(cyanomethyl)-4-(4-(6-methoxy-2,3-dihydro-1H-inden-1-yl)piperazin-1-yl)2-oxo-1,2 -dihydro-1,5-naphthyridine-3-carbonitrile

To a solution of 6-bromo-4-chloro-1-(cyanomethyl)-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (15 mg, 0.046 mmol) in DMF (0.5 ml) Hunig's base (0.024 mL, 0.139 mmol) and 1-(6-methoxy2,3-dihydro-1H-inden-1-yl)piperazine (16.16 mg, 0.070 mmol) were added. This reaction mixture was stirred by shaking overnight at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 32-72% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 97.0%; measured mass: 519.08; retention time: 2.06 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 97.6%; measured mass: 519.04; retention time: 1.36 min. The title compound (2.8 mg) was isolated in 11.7% yield.

Example 157 6-Bromo-1-(cyanomethyl)-4-(4-((4-fluorophenyl)(2-hydroxyphenyl)methyl)piperazin-1-yl)2-oxo-1,2-dihydro-1,5 -naphthyridine-3-carbonitrile

To a solution of 6-bromo-4-chloro-1-(cyanomethyl)-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (25 mg, 0.077 mmol) in DMF (0.5 ml) Hunig's base (0.040 mL, 0.232 mmol) and 2-((4fluorophenyl)(piperazin-1-yl)methyl)phenol, TFA (37.1 mg, 0.093 mmol) were added. This reaction mixture was stirred by shaking for 1 hour at room temperature. LC-MS analysis showed registration of the desired product. The resulting crude substance was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 36-76% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and

- 127 044500 was dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 573.08; retention time: 2.26 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 573.1; retention time: 1.58 min. The title compound (4.5 mg) was isolated in 10.2% yield.

Examples 158-160. 6-Bromo-1-(cyclopropylmethyl)-4-(4-((4-fluorophenyl)(2-methoxyphenyl)methyl)piperazin-1 -yl)-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile

To a solution of 6-bromo-4-chloro-1-(cyclopropylmethyl)-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (15 mg, 0.044 mmol) in DMF (1 ml) -((4-fluorophenyl)(2-methoxyphenyl)methyl)piperazine, TFA (18.36 mg, 0.044 mmol) and then Huenig's base (0.023 ml, 0.133 mmol) was added. This reaction mixture was stirred by shaking for 2 hours at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude substance was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-100% B for 20 min, then maintaining 100% B for 6 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 602.08; retention time: 1.78 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 602.13; retention time: 2.68 min. The title racemic compound (16.8 mg) was isolated in 63.4% yield.

The racemic material was further purified using SFC-chiral chromatography to obtain the compound of Example 159 (first eluting isomer) and the compound of Example 160 (second eluting isomer). Analytical LCMS was used to determine the final purity of the products.

Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration:

- 128 044500

MS and UV (220 nm).

Example 159. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 602.08;

retention time: 1.78 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 602.09; retention time: 2.68 min. Example 159 (6.6 mg) was isolated with the yield

24.9%.

Example 160. Analysis results for the 1st sample injection: purity: 100.0%; measured mass: 602.08; retention time: 1.78 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 602.11; retention time: 2.68 min. Example 160 (5.2 mg) was isolated in 19.6% yield.

Example 161 carbonitrile

To a solution of 6-bromo-4-chloro-1-(cyclopropylmethyl)-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (15 mg, 0.044 mmol) in DMF (15 ml) -(bis(4-fluorophenyl)methyl)piperazine, TFA (17.83 mg, 0.044 mmol) and then Huenig's base (0.023 ml, 0.133 mmol). This reaction mixture was stirred by shaking for 2 hours at room temperature. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 55-100% B for 20 min, then maintaining 100% B for 6 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 590.11; retention time: 2.66 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 590.1; retention time: 1.9 min. The title compound (14.5 mg) was isolated in 55.8% yield.

Examples 162-164. 6-Bromo-1 -(cyclopropylmethyl)-4-(4-( 1 -(4-fluorophenyl)ethyl)piperazin-1 -yl)-2oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile

To a solution of 6-bromo-4-chloro-1-(cyclopropylmethyl)-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (15 mg, 0.044 mmol) in DMF (15 ml), -(1-(4-fluorophenyl)ethyl)piperazine, TFA (14.28 mg, 0.044 mmol) and then Huenig's base (0.023 ml, 0.133 mmol). This reaction mixture was stirred by shaking for 2 hours at room temperature. According to LC-MS analysis, the reaction

- 129 044500 was completed. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-90% B for 20 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 510.07; retention time: 1.49 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 510.09; retention time: 2.38 min. The title racemic compound (14.8 mg) was isolated in 65.9% yield.

The racemic material was further purified using SFC-chiral chromatography to obtain the compound of Example 163 (first eluting isomer) and the compound of Example 164 (second eluting isomer). Analytical LCMS was used to determine the final purity of the products.

Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

Example 163. Analysis results for the 1st sample injection: purity: 97.5%; measured mass: 510.13; retention time: 2.37 min. Analysis results for the 2nd sample injection: purity: 98.3%; measured mass: 510.11; retention time: 1.51 min. Example 163 (3.5 mg) was isolated in 15.6% yield.

Example 164. Analysis results for the 1st sample injection: purity: 99.0%; measured mass: 510.13; retention time: 2.37 min. Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 510.11; retention time: 1.51 min. Example 164 (4.5 mg) was isolated in 20% yield.

General scheme for the preparation of the compound of example 204 and 4-(4-benzhydrylpiperazin-1-yl)-1-alkyl-3nitro-1,5-naphthyridin-2(1H)-one analogues

R.

A solution of 4-(4-benzhydrylpiperazin-1-yl)-3-nitro-1,5-naphthyridin-2(1H)-one in DMF (15.5 ml) was obtained. To each of the alkyl halides suspended in threaded tubes (16x100 mm), 0.500 ml of a solution of 4-(4-benzhydrylpiperazin-1-yl)-3-nitro-1,5-naphthyridin-2(1H)-one, potassium carbonate was added (25 mg, 0.18 mmol) and a magnetic stirrer armature. The tubes were capped and shaken for 4 hours at room temperature, then heated with stirring to 50°C overnight. The mixtures were transferred into empty 6 ml SPE cartridges for filtration, and the filtrate was collected in threaded tubes (16x48 mm). Each tube containing the reaction mixture was washed using 0.5 ml of DMF. The washes were transferred to filter cartridges. The resulting crude mixtures were purified by preparative HPLC using the conditions described in each example.

Example 165 4-(4-Benzhydrylpiperazin-1 -yl)-3-nitro-1 -(prop-2-yn-1 -yl)-1,5-naphthyridin-2(1H)-one

- 130 044500

The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 55-100% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters VEN C18, 2.0x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0% B, 0-100% B for 3 min, then maintain 100% B for 0.75 min; flow rate: 1 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters VEN C18, 2.0x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0% B, 0-100% B for 3 min, then maintain 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. The title compound was isolated in 54.7% yield (11.8 mg).

Example 166 1-(2-(1,3-Dioxan-2-yl)ethyl)-4-(4-benzhydrylpiperazin-1-yl)-3-nitro-1,5-naphthyridin-2(1H)-one

The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 70-100% B for 15 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters VEN C18, 2.0x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0% B, 0-100% B for 3 min, then maintain 100% B for 0.75 min; flow rate: 1 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters VEN C18, 2.0x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0% B, 0-100% B for 3 min, then maintain 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. The title compound was isolated in 36% yield (9 mg).

Example 176. 1 -Allyl-4-(4-benzhydrylpiperazin-1 -yl)-3-nitro-1,5-naphthyridin-2(1H)-one

- 131 044500

The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 70-100% B for 15 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters VEN C18, 2.0x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0% B, 0-100% B for 3 min, then maintain 100% B for 0.75 min; flow rate: 1 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters VEN C18, 2.0x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0% B, 0-100% B for 3 min, then maintain 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. The title compound was isolated in 37.4% yield (8.1 mg).

Example 177 4-(4-Benzhydrylpiperazin-1 -yl)-1-butyl-3-nitro-1,5-naphthyridin-2(1H)-one

The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 70-100% B for 15 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters VEN C18, 2.0x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0% B, 0-100% B for 3 min, then maintain 100% B for 0.75 min; flow rate: 1 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters VEN C18, 2.0x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0% B, 0-100% B for 3 min, then maintain 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. The title compound was isolated in 26.3% yield (5.9 mg).

Example 183 4-(4-(4-Benzhydrylpiperazin-1-yl)-3-nitro-2-oxo-1,5-naphthyridin-1(2H)-yl)butannitrile

- 132 044500

The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-90% B for 15 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. The title compound was isolated in 36.3% yield (8.3 mg).

Example 185

The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 70-100% B for 15 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters VEN C18, 2.0x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0% B, 0-100% B for 3 min, then maintain 100% B for 0.75 min; flow rate: 1 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters VEN C18, 2.0x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0% B, 0-100% B for 3 min, then maintain 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. The title compound was isolated in 22.7% yield (5.5 mg).

Example 186

- 133 044500

The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 70-100% B for 15 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters VEN C18, 2.0x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0% B, 0-100% B for 3 min, then maintain 100% B for 0.75 min; flow rate: 1 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters VEN C18, 2.0x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0% B, 0-100% B for 3 min, then maintain 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. The title compound was isolated in 32.6% yield (7.8 mg).

Example 187 4-(4-Benzhydrylpiperazin-1-yl)-3-nitro-1-(4-oxopentyl)-1,5-naphthyridin-2(1H)-one

The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 70-100% B for 15 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters VEN C18, 2.0x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0% B, 0-100% B for 3 min, then maintain 100% B for 0.75 min; flow rate: 1 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters VEN C18, 2.0x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0% B, 0-100% B for 3 min, then maintain 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. The title compound was isolated in 22% yield (5.2 mg).

Example 189 4-(4-Benzhydrylpiperazin-1-yl)-1-(3-(2-methoxyethoxy)propyl)-3-nitro-1,5-naphthyridin-2(1H)-one

- 134 044500

The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 65-100% B for 15 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters VEN C18, 2.0x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0% B, 0-100% B for 3 min, then maintain 100% B for 0.75 min; flow rate: 1 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters VEN C18, 2.0x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0% B, 0-100% B for 3 min, then maintain 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. The title compound was isolated in 32.3% yield (8.1 mg).

Example 190

The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 65-100% B for 15 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters VEN C18, 2.0x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0% B, 0-100% B for 3 min, then maintain 100% B for 0.75 min; flow rate: 1 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters VEN C18, 2.0x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0% B, 0-100% B for 3 min, then maintain 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. The title compound was isolated in 32.9% yield (7.6 mg).

Example 191 4-(4-Benzhydrylpiperazin-1-yl)-6-bromo-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine3-carbonitrile

- 135 044500

To a solution of 6-bromo-4-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (100 mg, 0.335 mmol) and Huenig's base (0.117 ml, 0.670 mmol) in DMF (5 ml) 1-benzhydrylpiperazine (85 mg, 0.335 mmol) was added in one go under nitrogen atmosphere. The progress of the reaction was monitored using HPLC. After 2 hours the reaction was completed. The reaction mixture was diluted with chloroform and washed (2x) with saturated aqueous NaHCO ₃ solution. The organic layer was then dried over MgSO ₄ , filtered and evaporated under reduced pressure overnight to give the desired product as a yellowish solid (132 mg, 77%). LC-MS conditions: injection sample volume: 3 µl; initial %B: 2; final % B: 98; gradient duration: 1.5 min; flow rate: 0.8 ml/min; wavelength: 220 nm; solvent pair: water/acetonitrile/TFA; solvent A: 100% water/0.05% TFA; solvent B: 100% acetonitrile/0.05% TFA; column: Waters Aquity VEN C18 2.1x50 mm, 1.7 µm MW1; oven temperature: 40. LC-MS results: 0.990 min. (M+H)+; 514. 1H NMR (400 MHz, chloroform-d) δ 7.62 (d, J=8.8 Hz, 1H), 7.53-7.46 (m, 5H), 7.32 (t, J=7.6 Hz, 4H), 7.25-7.20 ( m, 2H), 4.35 (s, 1H), 4.08-3.95 (m, 4H), 3.61 (s, 3H).

Example 192

To a solution of 6-bromo-4-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (100 mg, 0.335 mmol) and triethylamine (0.140 ml, 1.005 mmol) in DMF (2 ml) was added 5-fluoro-2-(piperazin-1ylmethyl)phenol 2,2,2-trifluoroacetate (109 mg, 0.335 mmol) and the resulting mixture was stirred at room temperature over the weekend. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; gradient: 10-60% B for 23 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. The product yield was 9.1 mg and its purity, determined by LC-MS analysis, was 96%. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. 1H NMR (500 MHz, DMSO) δ 7.95-7.92 (m, 1H), 7.88-7.85 (m, 1H), 7.20 (t, J=8.1 Hz, 1H), 6.63-6.57 (m, 2H), 3.91- 3.85 (m, 4H), 3.67 (s, 2H), 3.53 (s, 3H), 2.74-2.69 (m, 4H).

Example 193 6-Chloro-4-(4-(4-fluoro-2-hydroxybenzyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydro1,5-naphthyridine-3-carbonitrile

- 136 044500

To a solution of 4,6-dichloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (10 mg, 0.039 mmol) and triethylamine (0.016 ml, 0.118 mmol) in DMF ( 2 ml) 5-fluoro-2-(piperazin-1ylmethyl)phenol (8.28 mg, 0.039 mmol) was added and the resulting mixture was stirred overnight at room temperature. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 35-75% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. The resulting substance was further purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; gradient: 0-40% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis results for the 2nd sample injection: 1.32 min, [M+H]: 428.05. Analysis results for the 1st sample injection: 2.04 min, [M+H]; 428.04. ^1H NMR (500 MHz, DMSO^) δ 8.10 (d, J=8 Hz, 1H), 7.82 (d, J=8 Hz, 1H), 7.42 (br. s., 1H), 6.74 (d, J =9.5 Hz, 2H), 4.25 (br. s., 2H), 4.14-3.91 (m, 2H), 3.57 (s, 3H), 2.55 (s, 2H).

Example 194

To a solution of 5-fluoro-2-(piperazin-1-ylmethyl)phenol 2,2,2-trifluoroacetate (19.46 mg, 0.060 mmol) and 4-chloro-1,6-dimethyl-2-oxo-1,2 -dihydro-1,5-naphthyridine-3-carbonitrile (14.02 mg, 0.06 mmol) in DMF (2 ml), Huenig's base (10.48 μl, 0.060 mmol) was added and the resulting mixture was stirred overnight at room temperature in a nitrogen atmosphere. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 30-70% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. The product yield was 6.6 mg (27%) and its purity, determined by LC-MS analysis, was 99%. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) aceto

- 137 044500 nitrile: water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 98.8%; measured mass: 408.01; retention time: 1.21 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 408.02; retention time: 1.8 min. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ 7.91 (d, J=8.8 Hz, 1H), 7.59 (d, J=8.8 Hz, 1H), 7.20 (t, J=7.5 Hz, 1H), 6.64 -6.58 (m, 2H), 3.91 (br. s., J=3.3 Hz, 4H), 3.66 (s, 2H), 2.70 (br. s., 4H), 2.54 (s, 3H).

Example 195 -naphthyridine-3-carbonitrile

Potassium carbonate (82 mg, 0.590 mmol) was suspended in anhydrous DMF (4 ml). 4,6-Dichloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (50 mg, 0.197 mmol) and 1-(1-(2(allyloxy)- 4-fluorophenyl)ethyl)piperazine (52.0 mg, 0.197 mmol) and the resulting mixture was stirred at room temperature under nitrogen atmosphere overnight. The mixture was filtered and concentrated under high vacuum. The residue was dissolved in DMF and this crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 42-82% B for 19 min, then maintain 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. The product yield was 6.5 mg and its purity, determined by LC-MS analysis, was 100%. Analytical LC-MS was used to determine product purity. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 482.08; retention time: 1.54 min. 1H NMR (500 MHz, DMSO-d ₆ ) δ 8.04 (d, J=9.2 Hz, 1H), 7.78 (d, J=8.8 Hz, 1H), 7.43 (s, 1H), 6.89 (dd, J=11.6 , 2.4 Hz, 1H), 6.79 (s, 1H), 6.15-5.95 (m, 1H), 5.44 (s, 1H), 5.28 (d, J=10.6 Hz, 1H), 4.61 (br. s., 2H ), 4.02-3.92 (m, 1H), 3.83 (br. s., 4H), 2.69-2.55 (m, 4H), 1.29 (d, J=6.6 Hz, 3H). The N-CH ₃ resonance was not clearly expressed due to the suppression of water signals used to obtain the spectrum.

Example 196 4-(4-Benzhydrylpiperazin-1-yl)-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3-carbonitrile

4-(4-Benzhydrylpiperazin-1-yl)-6-bromo-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (20 mg, 0.039 mmol) was suspended in THF ( 2 ml). Diethylzinc (0.093 mL, 0.093 mmol) and tetrakis(triphenylphosphine) palladium (0) (4.49 mg, 3.89 μmol) were then added and the reaction mixture was heated at 70° C. in a sealed tube overnight. At the beginning of heating, the suspension turned into a yellow solution, which darkened upon subsequent heating overnight. Then

- 138 044500 the mixture was cooled, filtered and evaporated to dryness and the residue was dissolved in DMF (2 ml). This crude solution was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 45-90% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. The product yield was 5.2 mg and its purity, determined by LC-MS analysis, was 99%. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 99.3%; measured mass: 436.17; retention time: 1.34 min.

Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 436.15; retention time: 2.17 min. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ 8.51 (d, J=2.9 Hz, 1H), 7.97 (d, J=8.8 Hz, 1H), 7.68 (dd, J=8.8, 4.4 Hz, 1H) , 7.50 (d, J=7.3 Hz, 4H), 7.33 (t, J=7.7 Hz, 4H), 7.25-7.12 (m, 2H), 4.40 (s, 1H).

Example 198 6-Chloro-4-(4-(2-hydroxybenzyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile

To a solution of triethylamine (0.050 ml, 0.362 mmol) and 4,6-dichloro-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile (23 mg, 0.091 mmol) in DMF (4 ml ) 2-(piperazin-1ylmethyl)phenol-2,2,2,-trifluoroacetate (27.7 mg, 0.091 mmol) was added and the reaction mixture was stirred overnight at room temperature. The resulting mixture was evaporated to dryness and then dissolved in DMF (2 ml) and this substance was fractionated by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; gradient: 10-60% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. The product yield was 10.6 mg and its purity, determined by LC-MS analysis, was 95%. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 410.04; retention time: 1.36 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 95.1%; measured weight: 410.09; retention time: 1.9 min. 1H NMR (500 MHz, DMSO-d ₆ ) δ 8.10 (d, J=9.2 Hz, 1H), 7.82 (d, J=8.8 Hz, 1H), 7.38 (d, J=7.3 Hz, 1H), 7.29 ( t, J=7.5 Hz, 1H), 6.96 (d, J=8.1 Hz, 1H), 6.90 (t, J=7.3 Hz, 1H), 4.29 (br. s., 2H), 4.05 (br. s. , 2H), 3.57 (s, 3H), 2.55 (s, 2H). 4 H'S of piperazine is lost due to suppression of the water signal.

Example 199 -3-carbonitrile

- 139 044500

4,6-Dichloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (20 mg, 0.079 mmol) and Huenig's base (0.030 ml, 0.173 mmol) were dissolved in DMF ( 2 ml). N-ethyl-N-methyl-3(piperazin-1-ylmethyl)aniline 2,2,2-trifluoroacetate (27.3 mg, 0.079 mmol) was then added and the reaction mixture was stirred at room temperature under a nitrogen atmosphere. After 4 hours, the reaction came to completion. The crude reaction product was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 38-78% B for 19 min, then maintain 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. The product yield was 2.2 mg and its purity, determined by LC-MS analysis, was 100%. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 451.13; retention time: 1.02 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 451.11; retention time: 2.19 min. The water suppression technique used to obtain the product spectrum prevented complete signal assignment and only aromatic signals could be interpreted. 1H NMR (DMSO-d ₆ ) δ 8.09 (br. d, J=8.8 Hz, 1H), 7.83 (br. d, J=8.8 Hz, 1H), 7.05-7.28 (m, 1H), 6.57-6.87 ( m, 3H). The title compound 2.2 was isolated in 6.2% yield.

Example 200 4-(4-(7-(Allyloxy)-5-fluoro-2,3-dihydro-1H-inden-1-yl)piperazin-1-yl)-6-chloro-1-methyl-2- oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile

Solution of 4,6-dichloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (54 mg, 0.213 mmol), 1-(7-(allyloxy)-5-fluoro -2,3-dihydro-1H-inden-1-yl)piperazine (58.7 mg, 0.213 mmol) and triethylamine (0.059 ml, 0.425 mmol) in DMF (5 ml) were stirred under nitrogen over the weekend. The solvent was then removed under vacuum and the residue was dissolved in dichloromethane. The organic layer was washed with 1.5M potassium phosphate buffer (2x) and then dried over MgSO ₄ , filtered and concentrated in vacuo to give 159 mg of crude product. A portion of this material (127 mg) was fractionated by flash chromatography using 1-5% methanol in dichloromethane as eluent and a silica gel column (24 g) (Biotage). Homogeneous fractions were combined and evaporated under reduced pressure to give the reaction product as a brown oil (73 mg, 87%). 32 mg of the above crude reaction product was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-90% B for 20 min, then maintain 100% B

- 140 044500 for 5 minutes; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. The product yield was 9.8 mg and its purity, determined by LC-MS analysis, was 100%. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 494.05; retention time: 1.41 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 494.05; retention time: 2.22 min. ^1H NMR (400 MHz, chloroform-d) δ 7.64 (d, J=8.8 Hz, 1H), 7.51 (d, J=9.0 Hz, 1H), 6.56 (d, J=7.8 Hz, 1H), 6.44 ( d, J= 11.0 Hz, 1H), 6.13-6.00 (m, 1H), 5.52-5.41 (m, 1H), 5.29 (d, J=10.5 Hz, 1H), 4.62-4.50 (m, 2H), 4.37 (d, J=7.1 Hz, 1H), 4.03-3.86 (m, 4H), 3.81-3.68 (m, 1H), 3.61 (s, 3H), 3.03 (dt, J=16.4, 8.3 Hz, 1H), 2.85-2.69 (m, 5H), 2.31-2.06 (m, 2H).

Example 201 6-Chloro-4-(4-(5-fluoro-7-hydroxy-2,3-dihydro-1H-inden-1-yl)piperazin-1-yl)-1-methyl2-oxo-1, 2-dihydro-1,5-naphthyridine-3-carbonitrile

To a solution of 4-(4-(7-(allyloxy)-5-fluoro-2,3-dihydro-1H-inden-1-yl)piperazin-1-yl)-6-chloro-1-methyl-2-oxo -1,2-dihydro-1,5-naphthyridine-3-carbonitrile (75 mg, 0.152 mmol) in THF (3 ml) under a nitrogen atmosphere was added a catalytic amount of tetrakis(triphenylphosphine) palladium (0) (3.51 mg, 3 .04 µmol). This slightly brown solution was stirred for 5 minutes, then sodium borohydride (8.62 mg, 0.228 mmol) was added and the reaction mixture was left to stir overnight at room temperature. The reaction mixture was then transferred to a preparative TLC silica gel plate and eluted using 30% acetone in hexanes. Two fractions were eluted from the plate, the fraction with the lower Rf value was further purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 40-80% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. The product yield was 3.4 mg and its purity, determined by LC-MS analysis, was 94%. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 95.6%; measured mass: 453.97; retention time: 2.09 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 94.2%; measured weight: 453.95; retention time: 1.27 min. 1H NMR (500 MHz, DMSO-d ₆ ) δ 8.04 (d, J=8.8 Hz, 1H), 7.78 (d, J=8.8 Hz, 1H), 6.53 (d, J=8.8 Hz, 1H), 6.41 ( d, J=10.6 Hz, 1H), 4.46 (s, 1H), 3.84 (br. s., 2H), 3.58 (br. s., 1H), 3.09-3.01 (m, 1H), 2.88 (br. s., 1H). s., 1H), 2.832.68 (m, 2H), 2.65-2.55 (m, 2H), 2.15-1.97 (m, 2H).

Example 202 4-(4-Benzhydrylpiperazin-1 -yl)-6-ethyl-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine3-carbonitrile

- 141 044500

4-(4-Benzhydrylpiperazin-1-yl)-6-bromo-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (20 mg, 0.039 mmol) was suspended in THF ( 2 ml). Diethylzinc (0.093 mL, 0.093 mmol) and tetrakis(triphenylphosphine)palladium (0) (4.49 mg, 3.89 μmol) were then added and the reaction mixture was heated at 70° C. in a sealed tube overnight. At the beginning of heating, the suspension turned into a yellow solution. Then the reaction mixture was cooled, filtered and evaporated to dryness. The residue was dissolved in DMF (2 ml) and this solution was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 45-90% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. The product yield was 7.3 mg and its purity, determined by LC-MS analysis, was 100%. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 464.16; retention time: 1.52 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: 95:5 acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 464.14; retention time: 2.4 min. Ή NMR (500 MHz, DMSO-d ₆ ) δ 7.90 (d, J=8.8 Hz, 1H), 7.57 (d, J=8.4 Hz, 1H), 7.50 (d, J=7.3 Hz, 4H), 7.33 ( t, J=7.7 Hz, 4H), 7.24-7.18 (m, 2H), 4.42 (s, 1H), 3.95 (br. s., 2H), 3.50 (s, ZN*), 3.43 (br. s. , 2H), 2.80 (q, J=7.5 Hz, 2H), 2.60-2.53 (m, 4H), 1.20 (t, J=7.5 Hz, 3H).

Example 204 4-(4-Benzhydrylpiperazin-1-yl)-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3carbonitrile

CH ₃ ί Υ τ

HaC^N^y^CN

OyO (204)

To a solution of 1,6-dimethyl-2-oxo-4-(piperazin-1-yl)-1,2-dihydro-1,5-naphthyridine-3-carbonitrile 2,2,2-trifluoroacetate (26 mg, 0.065 mmol ) and triethylamine (0.027 ml, 0.196 mmol) in DMF (2 ml) was added (bromomethylene)dibenzene (16.17 mg, 0.065 mmol). The resulting mixture was stirred at room temperature under nitrogen atmosphere overnight. One additional equivalent of (bromomethylene)dibenzene (16.17 mg, 0.065 mmol) was added along with 1 additional equivalent of triethylamine and the reaction mixture was left stirring at room temperature for an additional 48 hours. The crude reaction mixture was then fractionated by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 µm, mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-90% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. The product yield was 3.3 mg and its purity, determined by LCMS analysis, was 100%. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, times

- 142 044500 measures of particles 1.7 microns; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 450.14; retention time: 2.35 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 450.12; retention time: 1.53 min. 1H NMR (500 MHz, DMSO-d6) δ 7.87 (d, J=8.8 Hz, 1H), 7.55 (d, J=8.8 Hz, 1H), 7.51 (d, J=7.7 Hz, 4H), 7.33 (t , J=7.5 Hz, 4H), 7.257.19 (m, 2H), 4.45 (s, 1H), 3.96 (br. s., 4H), 3.51 (s, 3H), 2.60 (br. s., 4H ), 2.51 (s, 3H).

Example 208 4-(4-Benzhydrylpiperazin-1-yl)-1-methyl-2-oxo-6-vinyl-1,2-dihydro-1,5-naphthyridine3-carbonitrile

Mixture of 4-(4-benzhydrylpiperazin-1-yl)-6-bromo-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (30 mg, 0.058 mmol), potassium vinyl trifluoroborate (9.37 mg, 0.070 mmol) and PdCl ₂ (dppf) (4.27 mg, 5.83 μmol) were placed in a dry microwavable tube, which was then sealed. BuOH (2 ml) and TEA (0.012 ml, 0.087 mmol) were added and the tube was degassed and then filled with nitrogen (3x). The reaction mixture was heated in a microwave oven at 110°C for 2 hours, then left to cool and evaporated to dryness under vacuum. The residue was then dissolved in DMF (2 ml) and this solution was fractionated by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 5090% B for 20 min, then maintain 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. The product yield was 15.7 mg and its purity, determined by LC-MS analysis, was 100%. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 462.17, 462.17, 462.17; retention time: 2.34, 2.42, 2.49 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 462.19; retention time: 1.5 min. 1H NMR (DMSO-d ₆ ) δ 7.93 (d, J=8.8 Hz, 1H), 7.81 (d, J=8.8 Hz, 1H), 7.49 (d, J=7.7 Hz, 4H), 7.32 (t, J =7.7 Hz, 4H), 7.14-7.25 (m, 2H), 6.75-6.85 (m, 1H), 6.13 (d, J=18.0 Hz, 1H), 5.50 (d, J=11.7 Hz, 1H), 4.40 (s, 1H). The water suppression technique used to obtain the product spectrum made it difficult to detect piperazine and protons in N-Me.

Example 209

- 143 044500

To a solution of 1-(cyclohexyl(phenyl)methyl)piperazine 2,2,2-trifluoroacetate (16.12 mg, 0.043 mmol) and 4,6-dichloro-1-methyl-2-oxo-1,2-dihydro-1 ,5-naphthyridine-3-carbonitrile (10 mg, 0.039 mmol) in DMF (1 ml) was added triethylamine (0.016 ml, 0.118 mmol). This reaction mixture was stirred overnight at room temperature. The crude reaction mixture was then fractionated by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 60-100% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. The product yield was 3.7 mg and its purity, determined by LC-MS analysis, was 98%. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 98.5%; measured mass: 476.1; retention time: 1.72 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 99.2%; measured mass: 476.11; retention time: 2.81 min. The NMR spectrum could not be fully interpreted due to the concentration of the sample being studied being too low. ¹ H NMR (DMSO) δ 8.00 (d, J=9.2 Hz, 1H), 7.72 (d, J=8.8 Hz, 1H), 7.30-7.38 (m, 2H), 7.21 (br. d, J=7.3 Hz , 3H), 3.83 (b. t, J=5.0 Hz, 4H), 3.50 (s, 3H).

Example 210 -naphthyridine-3-carbonitrile

4,6-Dichloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (20 mg, 0.079 mmol) was dissolved in dry DMF (2 ml) and potassium carbonate (32) was added .6 mg, 0.236 mmol). (R)-1benzyl-2-((benzyloxy)methyl)piperazine (25.7 mg, 0.087 mmol) was added and the reaction mixture was stirred at 60° C. in a sealed tube over the weekend. The reaction mixture was left to cool and then filtered and the resulting crude product was fractionated by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 60-100% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. The product yield was 1.2 mg and its purity, determined by LC-MS analysis, was 100%. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for 1st input

- 144 044500 sample: purity: 100.0%; measured mass: 514; retention time: 1.8 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 514.04; retention time: 2.8 min.

Example 211 4-(4-(2-Aminobenzyl)piperazin-1-yl)-6-bromo-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile

To a solution of 6-bromo-1-methyl-2-oxo-4-(piperazin-1-yl)-1,2-dihydro-1,5-naphthyridine-3-carbonitrile bis(2,2,2-trifluoroacetate) ( 200 mg, 0.347 mmol) in DMF (5 ml) was added tert-butyl (2-formylphenyl)carbamate (92 mg, 0.416 mmol). This reaction mixture was stirred for 1 hour at room temperature, then sodium cyanoborohydride (65.4 mg, 1.041 mmol) was added and the reaction mixture was stirred overnight at room temperature. Methanol was then added, the resulting mixture was filtered, and the filtrate was fractionated by reverse phase preparative HPLC using CH ₃ OH-H ₂ O-TFA as the eluent. Homogeneous fractions were combined and concentrated in vacuo to give tert-butyl (2-((4-(6-bromo-3-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-4- yl)piperazin-1-yl)methyl)phenyl)carbamate as a yellow solid. This material was dissolved in dichloromethane (5 ml) and TFA (3 ml, 38.9 mmol) was added. This reaction mixture was stirred for 2 hours at room temperature and then concentrated in vacuo to give an orange oil. This substance was further purified by reverse phase preparative HPLC using CH ₃ OH-H ₂ O-TFA as the eluent. The homogeneous fractions were combined and concentrated in vacuo to obtain the TFA salt of the title compound as a light yellow solid (70 mg, yield: 29.6%). Analytical LC-MS conditions: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). LC-MS results: 2.3 min, 453.0 (M+H)+. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ 7.96-7.90 (m, 1H), 7.88-7.83 (m, 1H), 7.10-6.86 (m, 2H), 6.67 (d, J=7.7 Hz, 1H ), 6.54 (t, J=7.5 Hz, 1H), 3.88-3.81 (m, 4H), 3.52 (s, 3H), 3.0 (s, 2H), 2.56-2.64 (m, 4H).

Example 212 )phenyl)methanesulfonamide

To a solution of 4-(4-(2-aminobenzyl)piperazin-1 -yl)-6-bromo-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile bis(2, 2,2-trifluoroacetate) (12 mg, 0.018 mmol) and triethylamine (0.012 ml, 0.088 mmol) in dichloromethane (2 ml), methanesulfonyl chloride (2.74 μl, 0.035 mmol) was added. This reaction mixture was stirred overnight at room temperature, then concentrated and the residue was dissolved in DMF-methanol. This solution was fractionated by preparative reverse phase LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 36-76% B for 19 min, then maintain 100% B for 4 min; flow rate: 20 ml/min. Homogeneous fractions containing the desired product were combined and dried using a centrifugal evaporator to obtain the following:

- 145 044500 title compound (1.8 mg, yield: 18.3%). Analytical LC-MS conditions: column: Waters XBridge C18, 2.1 mm x 50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). LC-MS results: 1.8 min, 531.1 (M+H)+. Ή NMR (500 MHz, DMSO-d ₆ ) δ 7.99-7.92 (m, 1H), 7.91-7.82 (m, 1H), 7.43 (br. d, J=7.7 Hz, 1H), 7.39-7.28 (m, 2H), 7.15 (br.d, J=6.2 Hz, 1H), 3.95-3.70 (m, 6H), 3.54 (s, 3H), 3.12 (s, 3H), 2.81-2.65 (m, 4H).

Example 213 )phenyl)benzamide

To a solution of 4-(4-(2-aminobenzyl)piperazin-1 -yl)-6-bromo-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile bis(2, 2,2-trifluoroacetamide) (17 mg, 0.025 mmol) and triethylamine (0.017 ml, 0.125 mmol) in dichloromethane (2 ml) was added benzoyl chloride (4.34 μl, 0.037 mmol). This reaction mixture was stirred for 1 hour at room temperature, then concentrated in vacuo, the residue was dissolved in a mixture of DMF-methanol and this solution was fractionated by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm ; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 40-80% B for 23 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Homogeneous fractions containing the desired product were combined and dried using a centrifugal evaporator to obtain the title compound (7.5 mg, yield: 51.2%). Analytical LC-MS conditions: column: Waters XBridge C18, 2.1 mm x 50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). LC-MS results: 2.5 min, 557.1 (M+H) ⁺ . Ή NMR (500 MHz, DMSO-d ₆ ) δ 11.20 (br. s, 1H), 8.31 (br. d, J=8.1 Hz, 1H), 8.01 (br. d, J=7.0 Hz, 2H), 7.97 -7.91 (m, 1H), 7.90-7.83 (m, 1H), 7.69-7.50 (m, 3H), 7.44-7.26 (m, 2H), 7.12 (b. t, J=7.3 Hz, 1H), 3.85 (br. m, 6H), 3.53 (s, 3H), 2.74 (br. s, 4H).

Example 214 )phenyl)-3-phenylurea

To a solution of 4-(4-(2-aminobenzyl)piperazin-1 -yl)-6-bromo-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile bis(2, 2,2-trifluoroacetamide) (17 mg, 0.025 mmol) and triethylamine (0.017 ml, 0.125 mmol) in THF (2 ml) was added phenyl isocyanate (4.09 μl, 0.037 mmol). This reaction mixture was stirred for 1 hour at room temperature, then concentrated in vacuo. The residue was dissolved in a mixture of DMF and methanol. This solution was fractionated by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 37-77% B for 22 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Homogeneous fractions containing the desired product were combined and dried using a centrifugal evaporator to obtain the title compound (6.8 mg, yield: 45.2%). Analytical LC-MS conditions: column: Waters XBridge C18, 2.1 mmx50 mm,

- 146 044500 particle size 1.7 microns; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). LC-MS results: 2.4 min, 572.2 (M+H)+. 1H NMR (500 MHz, DMSO-d ₆ ) δ 9.07 (s, 1H), 8.81 (br. s, 1H), 7.98-7.90 (m, 2H), 7.89-7.80 (m, 1H), 7.50 (d, J=7.7 Hz, 2H), 7.37-7.18 (m, 4H), 7.07-6.84 (m, 2H), 3.91 (br. s, 4H), 3.65 (br. s, 2H), 3.52 (s, 3H) , 2.65 (br. s, 4H).

Example 215. No. (2-((4-(6-Bromo-3-cyano-1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)piperazin1 -yl)methyl) phenyl)acetamide

To a solution of 4-(4-(2-aminobenzyl)piperazin-1-yl)-6-bromo-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile bis(2, 2,2-trifluoroacetamide) and triethylamine (0.012 ml, 0.088 mmol) (12 mg, 0.018 mmol) in dichloromethane (2 ml) was added acetyl chloride (1.878 μl, 0.026 mmol). This reaction mixture was stirred overnight at room temperature, then concentrated in vacuo and the residue was dissolved in a mixture of DMF and methanol. This solution was fractionated by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 26-76% B for 19 min, then maintain 100% B for 4 min; flow rate: 20 ml/min. Homogeneous fractions containing the desired product were combined and dried using a centrifugal evaporator to obtain the title compound (4.2 mg, yield: 45.7%). Analytical LC-MS conditions: column: Waters XBridge C18, 2.1 mm x 50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). LC-MS results: 2.1 min, 495.1 (M+H) ⁺ . 1H NMR (500 MHz, DMSO-d ₆ ) δ 10.15 (br. s, 1H), 7.97-7.89 (m, 2H), 7.88-7.81 (m, 1H), 7.38-7.19 (m, 2H), 7.07 ( t, J=7.5 Hz, 1H), 3.90 (br. s, 4H), 3.67 (s, 2H), 3.52 (s, 3H), 2.61-2.71 (br. m, 4H), 2.13 (s, 3H) .

Example 216 6-Chloro-4-(4-(indolin-7-ylmethyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridin-3-carbonitrile

To a solution of 6-chloro-1-methyl-2-oxo-4-(piperazin-1-yl)-1,2-dihydro-1,5-naphthyridine-3-carbonitrile bis(2,2,2-trifluoroacetate) ( 20 mg, 0.038 mmol) in DMF (1.5 ml) was added tert-butyl 7-formylindoline-1-carboxylate (13.95 mg, 0.056 mmol). This reaction mixture was stirred for 1 hour at room temperature. Sodium cyanoborohydride (7.09 mg, 0.113 mmol) was added and the reaction mixture was stirred overnight at room temperature. Methanol was then added, the resulting mixture was filtered, and the filtrate was fractionated by preparative reverse phase HPLC using CH3OH-H ₂ O-TFA as the eluent. Homogeneous fractions were collected and concentrated under reduced pressure overnight to give tert-butyl 7-((4-(6-chloro-3-cyano-1-methyl-2-oxo1,2-dihydro-1,5-naphthyridine-4 -yl)piperazin-1-yl)methyl)indoline-1-carboxylate as a yellow solid. This material was dissolved in dichloromethane (2 ml) and TFA (1 ml, 12.98 mmol) was added. This reaction mixture was stirred overnight at room temperature and then concentrated under reduced pressure. The residue was then fractionated by preparative LCMS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonite

- 147 044500 ril: water with 10 mM ammonium acetate; gradient: 38-78% B for 19 min, then maintain 100% B for 4 min; flow rate: 20 ml/min. Homogeneous fractions were combined and evaporated in vacuo to give the title compound (5.0 mg, yield: 29%). Analytical LC-MS conditions: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). LC-MS results: 1.9 min, 435.0 (M+H) ⁺ . 1H NMR (500 MHz, DMSO) δ 8.04 (d, J=9.2 Hz, 1H), 7.77 (d, J=8.8 Hz, 1H), 6.98 (d, J=6.6 Hz, 1H), 6.87 (d, J =7.3 Hz, 1H), 6.53 (t, J=7.3 Hz, 1H), 4.00-3.75 (m, 4H), 3.56-3.47 (m, 4H), 3.45 (s, 3H), 2.99-2.90 (m, 2H), 2.58-2.66 (br. m, 4H).

Example 217 8-(4-(bis(4-Fluorophenyl)methyl)piperazin-1-yl)-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2,7-dicarbonitrile

4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-6chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3 was added to a microwave test tube -carbonitrile (30 mg, 0.059 mmol), zinc (0.775 mg, 0.012 mmol), zinc cyanide (4.18 mg, 0.036 mmol) and complex of 1,1'-bis(diphenylphosphino)ferrocenepalladium(II) dichloride and dichloromethane (4 .84 mg, 5.93 µmol). The vessel was hermetically sealed, degassed three times successively and filled with nitrogen. NMP (2 ml) was added and the reaction mixture was heated for 1 hour at 75°C. The reaction mixture was then cooled, acetonitrile was added and the resulting mixture was filtered. The filtrate was fractionated by reverse phase preparative HPLC using acetonitrile-water-ammonium acetate as the eluent. Homogeneous fractions were combined and concentrated in vacuo overnight to obtain the title compound as a light yellow solid (22.3 mg, yield: 72.0%). Analytical LC-MS conditions: column: Phenomenex LUNA C18, 2.0x50 mm, particle size 3 µm; mobile phase A: (10:90) methanol:water with 0.1% TFA; mobile phase B: (90:10) methanol:water with 0.1% TFA; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: 3.3 min, 497.1(M+H) ⁺ . 1H NMR (400 MHz, DMSO-d ₆ ) δ 8.28-8.22 (m, 1H), 8.17-8.12 (m, 1H), 7.53 (dd, J=8.6, 5.6 Hz, 4H), 7.16 (t, J= 8.8 Hz, 4H), 4.54 (s, 1H), 3.91 (br. s., 4H), 2.56 (br. s., 4H).

The compounds of the following examples were prepared from suitable starting materials using procedures similar to those already described.

Example 218

Analytical LC-MS conditions: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. LC-MS results: 2.4 min, 506.0 (M+H)+. 1H NMR (500 MHz, DMSO^) δ 8.04 (d, J=9.2 Hz, 1H), 7.78 (d, J=8.8 Hz, 1H), 7.52 (dd, J=8.6, 5.7 Hz, 4H), 7.16 ( t, J=8.8 Hz, 4H), 4.51 (s, 1H), 3.88 (br. s., 4H), 3.51 (s, 3H), 2.56-2.53 (m, 4H).

Example 219 3-carbonitrile

- 148 044500

LC-MS (LC-2 Method): 2.1 min, 486.1 (M+H)+. 1H NMR (500 MHz, DMSO-d ₆ ) δ 8.06 (d, J=8.8 Hz, 1H), 7.79 (d, J=9.2 Hz, 1H), 7.54-7.46 (m, 2H), 7.42-7.35 (m , 1H), 7.33 (t, J=7.7 Hz, 2H), 7.26-7.17 (m, 1H), 7.10-6.97 (m, 1H), 6.84-6.73 (m, 2H), 4.78 (s, 1H), 3.90 (br. s, 4H), 3.52 (s, 3H), 2.69-2.56 (m, 4H).

Example 220 1,5-naphthyridine-3-carbonitrile

Analytical LC-MS conditions: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. LC-MS results: 2.1 min, 461.1 (M+H)+. 1H NMR (500 MHz, DMSO-de) δ 8.05 (d, J=9.2 Hz, 1H), 7.79 (d, J=9.2 Hz, 1H), 7.45-7.33 (m, 2H), 7.12 (t, J= 7.5 Hz, 1H), 7.03 (d, J=7.3 Hz, 1H), 6.65 (d, J=2.6 Hz, 1H), 4.21 (q, J=7.1 Hz, 2H), 3.90-3.74 (m, 6H) , 3.52 (s, 3H), 2.68 (br. s., 4H), 1.37 (t, J=7.2 Hz, 3H).

Example 221

Analytical LC-MS conditions: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. LC-MS results: 2.3 min, 444.1 (M+H)+. 1H NMR (500 MHz, DMSO-d ₆ ) δ 8.36 (d, J=8.8 Hz, 1H), 8.06 (d, J=8.8 Hz, 1H), 7.94 (d, J=8.4 Hz, 1H), 7.88 ( d, J=8.8 Hz, 1H), 7.80 (d, J=8.8 Hz, 1H), 7.62-7.44 (m, 4H), 4.00 (s, 2H), 3.83 (br. s., 4H), 3.53 ( s, 3H), 2.72 (br. s., 4H).

Example 222 5-naphthyridine-3-carbonitrile

- 149 044500

Analytical LC-MS conditions: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. LC-MS results: 2.4 min, 522.1 (M+H)+. ^1H NMR (500 MHz, DMSOL6) δ 8.06 (d, J=9.2 Hz, 1H), 7.79 (d, J=8.8 Hz, 1H), 7.49 (dd, J=8.6, 5.7 Hz, 2H), 7.43- 7.34 (m, 1H), 7.16 (t, J=8.8 Hz, 2H), 6.72-6.53 (m, 2H), 4.79 (s, 1H), 3.89 (br. s., 4H), 3.52 (s, 3H ), 2.62 (br. s., 2H), 2.57-2.51 (m, 2H).

Example 223 8-(4-Benzhydrylpiperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridin2-carbonitrile

8-Chloro-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile 0.22 ethyl acetate (94 mg, 0.331 mmol) was dissolved in DMF (3.3 ml ) until a solution is obtained. A magnetic stir bar, 1-(diphenylmethyl)piperazine (89 mg, 0.353 mmol) and then potassium carbonate (91 mg, 0.662 mmol) were added to the solution. This reaction mixture was placed under a nitrogen atmosphere and stirred at room temperature over the weekend. Inorganic salts were sucked out from the reaction solution with a pipette. A few drops of water and TFA (0.076 mL, 0.993 mmol) were added and the mixture was diluted to 6 mL using DMF and filtered through a 0.45 μm syringe filter, collecting the filtrate into three 2 mL sample tubes. The sample was purified by reverse phase HPLC using the following conditions: preparative HPLC system (Shimadzu) with discovery software: column: Waters Sunfire C18, 19 mmx150 mm; flow rate: 25 ml/min; solvent A (% composition): 10% acetonitrile - 90% water - 0.1% TFA; solvent B (% composition): 90% acetonitrile - 10% water - 0.1% TFA; registration: UV, 220 nm; gradient: from 20% B to 100% B for 20 min, maintaining 100% B for 5 min. Product retention time = 6.48 min to 7.31 min. The desired product was collected and the solvent was removed in vacuo to give the title compound (156 mg) as a yellow solid. LC-MS: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: 100% water with 0.05% trifluoroacetic acid; mobile phase B: 100% acetonitrile with 0.05% trifluoroacetic acid; temperature: 40°C; gradient: 2-98% B for 1.5 min, then maintain 98% B for 0.5 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. Retention time = 0.96 min; observed adducts: [M+H]; measured masses: 481.2. 1H NMR (acetonitrile-dz) δ 8.00-8.04 (m, 1H), 7.94-8.00 (m, 1H), 7.69 (d, J=7.6 Hz, 4H), 7.407.47 (m, 4H), 7.32-7.39 (m, 2H), 5.08 (br. s., 1H), 3.71 (t, J=4.5 Hz, 4H), 3.60 (s, 3H), 3.15 (br. s., 4H).

Example 224 3-Bromo-4-(4-(1-(4-fluorophenyl)ethyl)piperazin-1-yl)-1,6-dimethyl-1,5-naphthyridine2(1H)-oh

- 150 044500

DMF (0.7 ml) and 3-bromo-1,6-dimethyl-4-(piperazine- 1-yl)-1,5-naphthyridin-2(1H)-one (30.6 mg, 0.091 mmol) and then potassium carbonate (25.6 mg, 0.185 mmol) was added. This reaction mixture was stirred in a capped test tube for 4.5 hours at room temperature. According to HPLC analysis, the reaction was complete. Volatile components were removed from the reaction mixture under vacuum using a rotary evaporator. This crude reaction product was partitioned between ethyl acetate and saturated sodium bicarbonate solution. The organic extract was washed with saline and dried over magnesium sulfate. The drying agent was removed by filtration. Removal of the solvent from the filtrate gave the crude product as an amber oil (31 mg). The Rf value of the resulting product on a silica gel plate was ~0.26 (20% ethyl acetate in chloroform). The sample was dissolved in DMF/acetonitrile and the crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 30-78% B for 20 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. In the NMR spectrum shown, the effects caused by the suppression of water signals are not corrected. 1H NMR (DMSO-d6) δ 7.83 (d, J=8.9 Hz, 1H), 7.37 (br. dd, J=8.2, 5.8 Hz, 2H), 7.13 (br. t, J=8.9 Hz, 2H), 3.58 (s, 2H), 3.51 (br. t, J=4.4 Hz, 3H), 2.59 (br. s, 2H), 2.51 (br. s, 4H), 1.32 (br. d, J=6.7 Hz, 3H). Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 99.4%; measured mass: 458.92; retention time: 1.91 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 99.3%; measured mass: 459.04; retention time: 1.19 min.

Example 225 1,5-naphthyridine-3-carbonitrile

6-Bromo-4-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (15 mg, 0.050 mmol) was dissolved in DMF (0.5 ml) to obtain a solution . Then 1-ethyl-4-(piperazin-1-ylmethyl)1H-indole, 2 HCl (16.5 mg, 0.052 mmol) was added and then potassium carbonate (31 mg, 0.224 mmol) was added. This reaction mixture was placed under a nitrogen atmosphere for 1.75 hours at room temperature. Acetic acid (20 μL, 0.349 mmol) was added to the reaction mixture. The reaction mixture was diluted to a volume of 1.2 ml using acetonitrile and filtered through a syringe filter. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 4585% B for 18 min, then maintain 100% B for 4 min; flow rate: 20 ml/min. Tailcoat

- 151 044500 tions containing the desired product were combined and dried using a centrifugal evaporator. 1H NMR (DMSO-d6) δ 7.89-7.95 (m, 1H), 7.81-7.88 (m, 1H), 7.30-7.41 (m, 2H), 7.07-7.15 (m, 1H), 7.03 (d, J= 6.6 Hz, 1H), 6.66 (d, J=2.6 Hz, 1H), 4.20 (q, J=7.2 Hz, 2H), 3.74-3.95 (m, 6H), 3.51 (s, 3H), 2.69 (br. s., 2H), 1.39 (t, J=7.0 Hz, 3H). Analytical LCMS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 505.01; retention time: 2.16 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 505.02; retention time: 1.50 min. Proton NMR: Signal intensities are reduced (integrated intensity) near the frequency used to suppress the water signal. 1H NMR (DMSO-d ₆ ) δ 7.89-7.95 (m, 1H), 7.81-7.88 (m, 1H), 7.30-7.41 (m, 2H), 7.077.15 (m, 1H), 7.03 (d, J =6.6 Hz, 1H), 6.66 (d, J=2.6 Hz, 1H), 4.20 (q, J=7.2 Hz, 2H), 3.74-3.95 (m, 6H), 3.51 (s, 3H), 2.69 (ush . s., 2H), 1.39 (t, J=7.0 Hz, 3H).

Example 226

4-Chloro-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (6.5 mg, 0.028 mmol) and 1(4,4'-difluorobenzhydryl)piperazine ( 8.8 mg, 0.031 mmol) was dissolved in DMF (0.3 ml) to obtain a solution. Potassium carbonate (8 mg, 0.058 mmol) was then added. This reaction mixture was stirred for 2 hours at room temperature. LC-MS analysis indicated approximately 80% conversion of the starting materials. Additional 1-(4,4'difluorobenzhydryl)piperazine (2.3 mg, 7.98 μmol) was added to the reaction mixture. The reaction vessel was closed with a lid. The reaction mixture was stirred overnight at room temperature. The reaction mixture was then diluted with acetonitrile and filtered through a syringe filter. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 48-88% B for 20 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 486.13; retention time: 2.36 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 486.16; retention time: 1.59 min. Proton NMR signal intensities near the frequency used to suppress the water signal are distorted and uncorrected. 1H NMR (DMSO-d6) δ 7.86 (d, J=8.8 Hz, 1H), 7.44-7.58 (m, 5H), 7.09-7.18 (m, 4H), 4.52 (s, 1H), 3.90-3.97 (m , 4H), 3.50 (s, 2H), 2.53-2.60 (m, 4H).

Example 227 oh

- 152 044500

4-(4-(bis(4-Fluorophenyl)methyl)piperazin-1-yl)-1-methyl-1,5-naphthyridin-2(1H)-one (25 mg, 0.056 mmol) was dissolved in dioxane (0. 5 ml) until a solution is obtained. Pyridine (0.023 mL, 0.280 mmol) was then added and then trifluoroacetic anhydride (0.016 mL, 0.112 mmol) was added. This reaction mixture was placed under a nitrogen atmosphere and stirred for 4 hours at room temperature. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 60-100% B for 18 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. (Purity: 95.0%; RT: 2.54; observed adducts: [M+H]; measured masses: 543.07) Analytical conditions for 2nd sample injection: Column: Waters Acquity UPLC VEN

C18, 2.1x50 mm, particle size 1.7 microns; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. (Purity: 100.0%; RT: 1.7; observed adducts: [M+H]; measured masses: 543.24). Proton NMR signal intensities near the frequency used to suppress the water signal are distorted and uncorrected. 1H NMR (DMSO-06) δ 8.54 (d, J=4.0 Hz, 1H), 8.01 (d, J=8.8 Hz, 1H), 7.70 (dd, J=8.8, 4.0 Hz, 1H), 7.49 (dd, J=8.3, 5.7 Hz, 4H), 7.14 (t, J=8.8 Hz, 4H), 4.48 (s, 1H), 3.52 (s, 3H).

Example 228 N)-he

4-(4-(bis(4-Fluorophenyl)methyl)πpiperazin-1-yl)-1,6-dimethyl-1,5-naphthyridin-2(1H)-one (25 mg, 0.054 mmol) was dissolved in dioxane ( 0.5 ml) until a solution is obtained. Pyridine (0.022 ml, 0.271 mmol) was then added and then trifluoroacetic anhydride (0.015 ml, 0.109 mmol) was added. This reaction mixture was placed under a nitrogen atmosphere and stirred for 5.5 hours at room temperature. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 60-100% B for 18 min, then maintain 100% B for 7 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LCMS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 93.9%; measured mass: 557.14; retention time: 2.63 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid

- 153 044500 acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 91.9%; measured mass: 557.14; retention time: 1.79 min. Proton NMR signal intensities near the frequency used to suppress the water signal are distorted and uncorrected. ¹ H NMR (DMSO) δ 7.87 (d, J=8.8 Hz, 1H), 7.54 (d, J=8.8 Hz, 1H), 7.47 (dd, J=8.6, 5.7 Hz, 4H), 7.12 (t, J =8.8 Hz, 4H), 4.48 (s, 1H), 3.49 (s, 2H), 3.29 (s, 3H), 2.51-2.54 (m, 4H).

Example 229 1,5-naphthyridine-3-carbonitrile

6-Bromo-4-chloro-2-oxo-1-(prop-2-yn-1-yl)-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (16 mg, 0.050 mmol) was dissolved in DMF (0.5 ml) to obtain a solution. Then 1-(4,4'-difluorobenzhydryl)piperazine (14.6 mg, 0.051 mmol) was added and then potassium carbonate (13.9 mg, 0.101 mmol) was added. This reaction mixture was placed under a nitrogen atmosphere and stirred for 3.5 hours at room temperature. Acetic acid (10 μL, 0.175 mmol) was added to the reaction mixture. The reaction mixture was then diluted to 1 ml using acetonitrile and filtered. The resulting crude material was purified by preparative LC-MS using the following conditions:

column: XBridge C18, 19x200 mm, particle size 5 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 55-95% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 573.96; retention time: 1.9 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 573.96; retention time: 2.55 min. Proton NMR signal intensities near the frequency used to suppress the water signal are distorted and uncorrected. 1H NMR (DMSO) δ 7.86-8.00 (m, 2H), 7.50 (dd, J=8.4, 5.9 Hz, 4H), 7.14 (t, J=8.8 Hz, 4H), 5.00 (d, J=1.8 Hz, 2H), 4.53 (s, 1H), 3.91 (br. s., 4H), 2.56 (br. s., 4H).

Example 230 4-(4-(bis(4-Fluorophenyl)methyl)piperazin-1-yl)-3-bromo-1-methyl-1,5-naphthyridin-2(1H)one

A solution was prepared by dissolving 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-1-methyl-1,5naphthyridin-2(1H)-oneα (26.5 mg, 0.059 mmol) in DMF (0.2 ml) followed by the addition of NBS (12.6 mg, 0.071 mmol). This reaction mixture was placed under a nitrogen atmosphere and stirred for 2.5 hours at room temperature. The reaction mixture was diluted to 1 ml using acetonitrile and then filtered through a syringe filter. The resulting crude material was purified

- 154 044500 by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-100% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. (Purity: 98.8%; RT: 2.61; observed adducts: [M+H]; measured masses: 524.99). Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. (Purity: 100.0%; RT: 1.44; observed adducts: [M+H]; measured masses: 525.02). Proton NMR signal intensities near the frequency used to suppress the water signal are distorted and uncorrected. 1H NMR (DMSO-d ₆ ) δ 8.54 (d, J=4.4 Hz, 1H), 7.98 (d, J=8.8 Hz, 1H), 7.63 (dd, J=8.6, 4.2 Hz, 1H), 7.51 (dd , J=8.4, 5.5 Hz, 4H), 7.14 (t, J=8.8 Hz, 4H), 4.46 (s, 1H), 3.63 (s, 3H), 3.56 (t, J=5.0 Hz, 4H).

Example 231 naphthyridine-2-carboxamide

8-(4-(bis(4-Fluorophenyl)methyl)piperazin-1-yl)-7-cyano-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carboxylic acid ( 15 mg, 0.029 mmol) and aza-HOBt (5.0 mg, 0.037 mmol) were dissolved in DMF (0.3 ml) to obtain a solution. DIEA (5.4 μL, 0.031 mmol) was added to the reaction mixture, and then EDC (9.1 mg, 0.047 mmol) was added. This reaction mixture was stirred for 5 min at room temperature and then dimethylamine (29.1 μl, 0.058 mmol) was added. The reaction vessel was capped and the reaction mixture was stirred for 2 hours at room temperature. Acetic acid (7 μL, 0.122 mmol) was added and the reaction mixture was then diluted with acetonitrile. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 38-78% B for 20 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 543.18; retention time: 1.41 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 543.24; retention time: 2.01 min. Proton NMR signal intensities near the frequency used to suppress the water signal are distorted and uncorrected. 1H NMR (DMSO-d ₆ ) δ 8.04 (d, J=9.2 Hz, 1H), 7.80 (d, J=8.8 Hz, 1H), 7.49 (dd, J=8.6, 5.7 Hz, 4H), 7.14 (t , J=8.8 Hz, 4H), 4.55 (s, 1H), 3.84-3.95 (m, 4H), 3.54 (s, 3H), 3.00 (br. s, 2H), 2.91 (br. s, 2H), 2.52-2.57 (m, 4H).

Example 232 4-(4-(bis(4-Fluorophenyl)methyl)piperazin-1-yl)-1,6-dimethyl-3-(trifluoromethyl)-1,5

- 155 044500 naphthyridin-2(1H)-one

References: Togni et al. J. Fluorine Chem. 131 (2010) 951-957. Onset observed >135°C; Haller, J. Org. Process Res. Dev., 2013, 17 (3), pp. 318-319.

4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-1,6-dimethyl1,5-naphthyridine was added to a 1 dram high pressure test tube containing a new Teflon magnetic stirrer armature. -2(III)-one (15 mg, 0.033 mmol) and chlorotris(trimethylsilyl)silane (16.2 mg, 0.057 mmol) followed by dissolution in acetonitrile (0.35 ml). 1-Trifluoromethyl-3,3-dimethyl1,2-benziodoxole (21.5 mg, 0.065 mmol) was then added. The tube was sealed with a seal cap and heated in an oil bath preheated to 80°C behind a blast shield for 24 hours. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 60-100% B for 22 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LCMS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 529.16; retention time: 2.67 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 529.22; retention time: 1.71 min. Proton NMR signal intensities near the frequency used to suppress the water signal are distorted and uncorrected. ¹ H NMR (DMSO-d ₆ ) δ 7.85 (d, J=8.8 Hz, 1H), 7.50 (dt, J=8.5, 5.6 Hz, 5H), 7.13 (t, J=8.8 Hz, 4H), 4.49 ( s, 1H), 3.67 (br. s, 4H), 3.50 (s, 3H), 2.52-2.58 (m, 4H).

Example 233 4-(4-(bis(4-Fluorophenyl)methyl)piperazin-1-yl)-3-chloro-1,6-dimethyl-1,5-naphthyridin2(1H)-one

4-(4-(bis(4-Fluorophenyl)methyl)piperazin-1-yl)-1,6-dimethyl-1,5-naphthyridin-2(III)-one (15 mg, 0.033 mmol) was dissolved in DMF ( 0.33 ml) until a solution is obtained. This solution was cooled to 0°C and then n-chlorosuccinimide (5.0 mg, 0.037 mmol) was added. This reaction mixture was placed under a nitrogen atmosphere, stirred at 0°C for one hour, then removed from the ice bath, warmed to room temperature and stirred for an additional hour. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-100% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for 1

- 156 044500 th sample input: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 495.1; retention time: 1.53 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 495.09; retention time: 2.49 min. Proton NMR signal intensities near the frequency used to suppress the water signal are distorted and uncorrected. 1H NMR (DMSO-d ₆ ) δ 7.84 (d, J=8.4 Hz, 1H), 7.40-7.54 (m, 5H), 7.13 (t, J=8.8 Hz, 4H), 4.48 (s, 1H), 3.54 -3.64 (m, 6H), 2.54 (s, 3H).

Example 234

4-(4-(bis(4-Fluorophenyl)methyl)piperazin-1-yl)-1,6-dimethyl-1,5-naphthyridin-2(1H)-one (20 mg, 0.043 mmol) was dissolved in acetonitrile ( 0.4 ml) until a solution is obtained. This solution was cooled to 0°C and 1-chloromethyl-4-fluoro-1,4-diazoniabicyclo[2.2.2]octane bis(tetrafluoroborate) (15.39 mg, 0.043 mmol) dissolved in water (0.15 ml) was added ) and THF (0.15 ml). This reaction mixture was placed under a nitrogen atmosphere, stirred at 0°C, warmed to room temperature for 1.5 hours and then stirred for 3 hours. LC-MS analysis of the reaction mixture showed approximately 25% conversion of the starting materials to the reaction product . The reaction mixture was cooled to 0°C and additional 1-chloromethyl-4-fluoro-1,4-diazoniabicyclo[2.2.2]octane bis(tetrafluoroborate) (16.3 mg, 0.046 mmol) in water (0.2 ml) was added ). The reaction vessel was capped and the reaction mixture was stirred for 25 minutes at 0°C. The reaction vessel was removed from the ice bath. The reaction mixture was stirred for 35 minutes, warming to room temperature. LC-MS analysis showed approximately 50% conversion of the starting materials into the reaction product. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 45-85% B for 23 min, then maintain 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 96.4%; measured mass: 479.2; retention time: 2.42 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 96.3%; measured mass: 479.2; retention time: 1.48 min. Proton NMR signal intensities near the frequency used to suppress the water signal are distorted and uncorrected: 1H NMR (DMSO-d6) δ 7.81 (d, J=8.4 Hz, 1H), 7.48 (dd, J=8.6, 5.7 Hz, 4H ), 7.41 (d, J=8.8 Hz, 1H), 7.12 (t, J=8.8 Hz, 4H), 4.48 (s, 1H), 3.58 (br. s, 3H), 3.56 (s, 3H).

Example 235 TFU

- 157 044500

In a 1 dram test tube containing 6-bromo-1-methyl-2,4-dioxo-1,2,3,4-tetrahydro-1,5naphthyridine-3-carbonitrile (39.7 mg, 0.142 mmol) in acetonitrile ( 1.4 ml), DIEA (0.149 ml, 0.850 mmol) was added. This reaction mixture was stirred for several minutes and then tosyl chloride (81 mg, 0.425 mmol) was added. The reaction mixture was capped under a nitrogen atmosphere and stirred for 20 hours at room temperature. Half of the above volume of the reaction mixture (the reaction mixture was heterogeneous and amber in color) was added to a 1 dram test tube containing 1-(4,4'-difluorobenzhydryl)piperazine (26.6 mg, 0.092 mmol), acetonitrile (0.2 ml) and 1-methylpiperidine (0.011 ml, 0.092 mmol). This reaction mixture was capped under a nitrogen atmosphere and stirred for 3 hours at room temperature. After HPLC analysis, 1-(4,4'-difluorobenzhydryl)piperazine (20.47 mg, 0.071 mmol) was added to the reaction mixture. The reaction vessel was closed with a lid. The reaction mixture was stirred at room temperature for an additional 1.5 hours. Acetic acid (0.020 mL, 0.355 mmol) was added to the reaction mixture along with DMF and the mixture was filtered through a syringe filter. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; gradient: 15-55% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 97.4%; measured mass: 549.94; retention time: 2.5 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured weight: 549.95; retention time: 1.8 min. 1H NMR (DMSO-dg) δ 7.89-7.96 (m, 1H), 7.81-7.88 (m, 1H), 7.54 (dd, J=8.3, 5.7 Hz, 4H), 7.17 (t, J=8.8 Hz, 4H ), 4.73 (br. s., 1H), 3.91 (br. s., 4H), 3.51 (s, 3H), 2.68 (br. s., 4H).

Example 236

To a 1 dram test tube containing 3-bromo-4-hydroxy-1-methyl-1,5-naphthyridin-2(1H)-one (16.4 mg, 0.064 mmol), add acetonitrile (0.5 ml) and then DIEA (44.9 μL, 0.257 mmol) was added. Trifluoromethanesulfonic acid anhydride (13.04 μL, 0.077 mmol) was added to this reaction mixture. The reaction mixture was placed under a nitrogen atmosphere and stirred for 55 minutes at room temperature. HPLC analysis of the reaction mixture showed approximately 60% conversion of the starting materials into the reaction product. The reaction mixture was stirred under nitrogen atmosphere at room temperature overnight. Further monitoring using HPLC analysis indicated no significant change in the reaction composition. Trifluoromethanesulfonic acid anhydride (5.5 µl, 0.033 mmol) was added to the reaction mixture. The reaction mixture was stirred under nitrogen atmosphere for 45 minutes. The next HPLC analysis showed 91% conversion of the starting materials into the reaction product. Volatile components were removed from the reaction mixture under vacuum using a rotary evaporator. The resulting crude product was partitioned between ethyl acetate and 1.5 M potassium hydrogen orthophosphate solution. The organic extract was washed successively with 1.5 M potassium hydrogen orthophosphate solution and saline solution and dried over magnesium sulfate. The drying agent was removed by filtration. Solvents were removed in vacuo using a rotary evaporator to yield the intermediate, 3-bromo-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridin-4-yl trifluoromethanesulfonate, as a brown film (26 mg). LC-MS: column: Waters Acquity VEH C18 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: 100% water with 0.05% TFA; mobile phase B: 100% acetonitrile with 0.05% TFA; temperature: 40°C; gradient: from 2% B to 98% B for 1.5 min, then maintaining 98% B for 0.5 min; flow rate: 0.8 ml/min; registration: MS and UV (220 nm). Injected sample volume: 3 µl. Retention time = 1.01 min; observed adducts: [M+H]; measured masses: 386.85.

A solution of 3- bromo-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridin-4-yl trifluoromethanesulfonate (26 mg, 0.067 mmol) in DCM (0.45 ml). The tube was capped and the reaction mixture was stirred for 2 hours at room temperature. Volatile components were removed from the reaction mixture under vacuum using a rotary evaporator. The crude reaction product was dissolved in a mixture of DMF/acetonitrile (1:1, 1 ml). The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 40-80% B for 15 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator.

To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. (Purity: 100.0%; RT: 1.88; observed adducts: [M+H]; measured masses: 480.04). Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. (Purity: 100.0%; RT: 1.29; observed adducts: [M+H]; measured masses: 480.04). Proton NMR signal intensities near the frequency used to suppress the water signal are distorted and uncorrected. ¹ H NMR (DMSO-d ₆ ) δ 8.53 (d, J=2.9 Hz, 1H), 7.99 (d, J=7.7 Hz, 1H), 7.63 (dd, J=8.6, 4.2 Hz, 1H), 7.32- 7.44 (m, 2H), 7.11 (t, J=7.5 Hz, 1H), 7.03 (d, J=5.9 Hz, 1H), 6.65 (br. s., 1H), 4.20 (q, J=7.3 Hz, 2H), 3.79 (br. s., 1H), 3.64 (s, 3H), 3.53 (br. s., 2H), 2.63 (br. s., 2H), 1.37 (t, J=7.2 Hz, 3H ).

Example 237 6-Bromo-1-methyl-4-(4-(naphthalene-1-ylmethyl)piperazin-1-yl)-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile

6-Bromo-4-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (15 mg, 0.050 mmol) was dissolved in DMF (0.5 ml) to obtain a solution . Then 1-(naphthalene-1-ylmethyl)piperazine, 3 TFA (28.6 mg, 0.050 mmol) was added and then potassium carbonate (34.7 mg, 0.251 mmol) was added. This reaction mixture was placed under a nitrogen atmosphere and stirred for 2 hours at room temperature. The reaction mixture was then diluted with acetonitrile and 15 μL of acetic acid was added. The reaction mixture was then stirred and filtered through a syringe filter. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 50-90% B in those

- 159 044500 reading for 15 minutes, then maintaining 100% B for 5 minutes; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 487.9; retention time: 1.35 min. ¹ H NMR (DMSO-d ₆ ) δ 8.36 (d, J=9.2 Hz, 1H), 7.92 (d, J=9.2 Hz, 2H), 7.82-7.88 (m, 2H), 7.437.62 (m, 4H ), 4.00 (s, 2H), 3.83 (d, J=4.8 Hz, 4H), 3.52 (s, 3H), 2.67-2.77 (m, 4H).

Example 238 -3-carbonitrile, TFU

6-Bromo-4-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (15 mg, 0.050 mmol) was dissolved in DMF (0.5 ml) to obtain a solution . Then 1-([1,1'-biphenyl]-2-ylmethyl)piperazine, 2 TFA (24.14 mg, 0.050 mmol) was added and then potassium carbonate (29 mg, 0.210 mmol) was added. The reaction mixture was capped under a nitrogen atmosphere and stirred for 2 hours at room temperature. The reaction mixture was diluted using DMF and acetonitrile and two drops of water and 30 μl of acetic acid were added. The reaction mixture was heated until the solids dissolved and filtered through a syringe filter. After cooling, a very small amount of the substance precipitated, 15 μl of TFA was added to facilitate solubilization. This solution was filtered again through a syringe filter. The resulting crude substance was purified by preparative LCMS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; gradient: 15-55% B for 18 min, then maintain 100% B for 4 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 513.93; retention time: 2.42 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 514.18; retention time: 1.46 min. Proton NMR signal intensities near the frequency used to suppress the water signal are distorted and uncorrected. 1H NMR (DMSO-d ₆ ) δ 7.90-7.97 (m, 1H), 7.83-7.89 (m, 1H), 7.71 (d, J=7.3 Hz, 1H), 7.48 (t, J=7.3 Hz, 4H) , 7.35-7.43 (m, 3H), 7.28-7.34 (m, 1H), 4.08 (br. s., 1H), 3.89 (br. s., 3H), 3.51 (s, 2H), 2.93 (br. s., 1H).

Example 239 4-(4-(bis(4-Fluorophenyl)methyl)piperazin-1-yl)-3-bromo-1,6-dimethyl-1,5-naphthyridin2(1H)-one

- 160 044500

4-(4-(bis(4-Fluorophenyl)methyl)piperazin-1-yl)-1,6-dimethyl-1,5-naphthyridin-2(1H)-one (25 mg, 0.054 mmol) was dissolved in DMF ( 0.25 ml) until a solution is obtained. NBS (12.3 mg, 0.069 mmol) was then added. This reaction mixture was capped and stirred for 1.5 hours at room temperature. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 55-95% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 539.07; retention time: 2.58 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 539.06; retention time: 1.65 min. ¹ H NMR (chloroform-d) δ 7.55 (d, J=8.7 Hz, 1H), 7.39-7.47 (m, 4H), 7.30 (d, J=8.7 Hz, 1H), 6.96-7.05 (m, 4H) , 4.32 (s, 1H), 3.71 (s, 3H), 3.65-3.70 (m, 4H), 2.56-2.64 (m, 7H).

Example 240 Methyl 8-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-7-cyano-5-methyl-6-oxo-5,6dihydro-1,5-naphthyridine-2-carboxylate

In a 2 dram test tube containing methyl 7-cyano-5-methyl-6-oxo-8-(piperazin-1-yl)-5,6dihydro-1,5-naphthyridine-2-carboxylate, HCl (52 mg, 0.143 mmol), 1.5 ml of DCM and 1.5 ml of 1.0 M sodium carbonate solution were added. 4,4'-(bromomethylene)bis(fluorobenzene) (59.5 mg, 0.210 mmol) was then added. The reaction vessel was closed with a lid. The reaction mixture was stirred at room temperature for 2 days. The reaction mixture was then further diluted with DCM and saline. The resulting product was extracted with ethyl acetate. The organic extract was dried over magnesium sulfate, filtered and the solvent was removed under vacuum to obtain the crude reaction product. This crude mixture was purified by chromatography on silica gel, eluting first with DCM to remove non-polar impurities and then with 15% ethyl acetate in DCM. Fractions containing pure reaction product (TLC) were combined and the solvent was removed in vacuo to obtain the desired product as a beige solid. LC-MS: column: Waters Acquity VEN 2.1 mm x 50 mm, particle size 1.7 µm; mobile phase A: 10:90 acetonitrile:water with 0.1% TFA; mobile phase B: 90:10 acetonitrile:water with 0.1% TFA; temperature: 40°C; gradient: from 0% B to 100% B for 2 min, then maintaining 100% B for 1 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Injected sample volume: 1 µl.

Retention time = 1.90 min; observed adducts: [M+H]; measured masses: 530.20. LC-MS: column: Phenomenex Luna C18, 2 mmx50 mm, particle size 3 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate - 161 044500; temperature: 40°C; gradient: from 0% B to 100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: MS and UV (220 nm). Injected sample volume: 3 µl. Retention time = 3.43 min; observed adducts: [M+H]; measured masses: 530.21.

Example 241 8-(4-(bis(4-Fluorophenyl)methyl)piperazin-1-yl)-7-cyano-5-methyl-6-oxo-5,6-dihydro1,5-naphthyridine-2-carboxylic acid

CH ₃ °o (241)

Into a 1 dram test tube containing methyl 8-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)7-cyano-5-methyl-6-oxo-5,6-dihydro-1,5- naphthyridine-2-carboxylate (50 mg, 0.094 mmol), 0.5 ml THF was added to form a suspension. 0.5 ml DCM was then added to facilitate solubilization. Additional THF (0.45 mL) was added, causing even more starting material to precipitate out of solution. To solubilize the starting material, 0.1 ml of DCM was added in portions. An additional 0.4 ml of DCM was required to achieve solubilization. The reaction solution composition contained 0.9 ml DCM and 0.95 ml THF. Potassium trimethylsilanolate (15.7 mg, 0.122 mmol) was added to the reaction mixture, and the reaction mixture was capped and stirred for 4 hours at room temperature. Hydrochloric acid (1 M, 0.123 mL, 0.123 mmol) was then added to the reaction mixture and the reaction mixture was stirred for several minutes. Chloroform was added. The reaction mixture was transferred to a separatory funnel and additional chloroform was added along with water. After stirring, the organic phase was separated and the aqueous phase was extracted with chloroform. The organic extracts were combined, washed with saline and dried over magnesium sulfate. The drying agent was filtered off and the solvent was removed from the filtrate under vacuum using a rotary evaporator. The resulting crude material was purified by preparative reverse phase HPLC using a solvent system of acetonitrile/water/0.1% TFA and UV detection at 220 nm. The fractions were analyzed by LC-MS and the reaction product fraction was removed in vacuo by using a rotary evaporator to obtain the desired product as a yellowish solid. LC-MS: column: Waters Acquity VEN 2.1 mm x 50 mm, particle size 1.7 µm; mobile phase A: 10:90 acetonitrile:water with 0.1% TFA; mobile phase B: 90:10 acetonitrile:water with 0.1% TFA; temperature: 40°C; gradient: from 0% B to 100% B for 2 min, then maintaining 100% B for 1 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Injected sample volume: 1 µl. Results: retention time = 1.70 min; observed adducts: [M+H]; measured masses: 516.20. 1H NMR (acetonitrile-d ₃ ) δ 8.25 (d, J=8.8 Hz, 1H), 7.92 (d, J=9.0 Hz, 1H), 7.64-7.78 (m, 4H), 7.17 (t, J=8.7 Hz , 4H), 5.05 (br. s., 1H), 4.16 (br. s., 4H), 3.57 (s, 3H), 3.15 (br. s., 4H).

Example 242 4-[4-(Diphenylmethyl)piperazin-1-yl]-1-methyl-3-nitro-1,2-dihydro-1,5-naphthyridin-2one

The title compound was prepared according to the general procedure used for the synthesis of Example 165. The compound (1.8 mg) was isolated in 0.9% yield. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0100% B for 3 min, then maintain 100% B for 0.75 min; flow rate: 1.0 ml/min;

- 162 044500 registration: UV, 220 nm.

Example 243 4-[4-(Diphenylmethyl)piperazin-1-yl]-1-ethyl-3-nitro-1,2-dihydro-1,5-naphthyridin-2one

The title compound was prepared according to the general procedure used for the synthesis of Example 165. The compound (5 mg) was isolated in 23.7% yield. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0100% B for 3 min, then maintain 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Example 244 4-[4-(Diphenylmethyl)piperazin-1-yl]-1-(2-methoxyethyl)-3-nitro-1,2-dihydro-1,5-naphthyridin-2-one

The title compound was prepared according to the general procedure used for the synthesis of Example 165. The compound (10.1 mg) was isolated in 35.5% yield. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0100% B for 3 min, then maintain 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Example 245

The title compound was prepared according to the general procedure used for the synthesis of Example 165. The compound (5.9 mg) was isolated in 21.5% yield. For determining

- 163 044500 final product purity, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0100% B for 3 min, then maintain 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Example 246 Ethyl 2-{4-[4-(diphenylmethyl)piperazin-1-yl]-3-nitro-2-oxo-1,2-dihydro-1,5-naphthyridin-1-yl} acetate

The title compound was prepared according to the general procedure used for the synthesis of Example 165. The compound (7.5 mg) was isolated in 50.8% yield. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0100% B for 3 min, then maintain 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Example 248 4-[4-(Diphenylmethyl)piperazin-1-yl]-3-nitro-1-propyl-1,2-dihydro-1,5-naphthyridin2-one

The title compound was prepared according to the general procedure used for the synthesis of Example 165. The compound (9.5 mg) was isolated in 27.9% yield. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0100% B for 3 min, then maintain 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Example 250

- 164 044500

1,6-Dimethyl-2-oxo-4-(piperazin-1-yl)-1,2-dihydro-1,5-naphthyridine-3-carbonitrile, TFA (20 mg, 0.050 mmol), cyclopropyl(4-fluorophenyl )methanol (11.71 mg, 0.070 mmol), (cyanomethyl)trimethylphosphonium iodide (24.46 mg, 0.101 mmol) were mixed in propionitrile (252 μl) in a sealed 2 dram reaction vessel. Huenig's base (75 μl, 0.429 mmol) was added and the reaction mixture was heated for 4 hours at 110°C. According to HPLC analysis, the starting material was consumed. The reaction mixture was diluted with ethyl acetate (5 ml) and extracted with water (3 x). The organic fraction was concentrated under a nitrogen stream. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 200 mmx19 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: maintaining 30% B for 0 min, 30-70% B for 23 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min; Column temperature: 25°C. Fraction collection was triggered by MS and UV signals. Fractions containing the desired product were combined and dried using a centrifugal evaporator. The compound (1.8 mg) was isolated in 8.3% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 98.0%; measured mass: 432.16; retention time: 1.42 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 98.4%; measured mass: 432.2; retention time: 2.03 min.

Example 251 3-carbonitrile

H ₃ C. ^CH ₃

O N O -όΎυ..,,

The title compound was prepared according to the general procedure used for the synthesis of Example 250. The compound (23.2 mg) was isolated in 42.5% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 98.1%; measured mass: 546.16; retention time: 2.31 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 im). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 546.19; retention time: 1.51 min.

Example 252 3-carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 250. The compound (1.7 mg) was isolated in 9.2% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 92.7%; measured mass: 370.2; retention time: 1.02 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 95.3%; measured mass: 370.21; retention time: 1.37 min.

Example 253 4-[4-(3,4-Dihydro-2H-1-benzopyran-4-yl)piperazin-1-yl]-1,6-dimethyl-2-oxo-1,2-dihydro-1, 5-naphthyridine-3-carbonitrile

CH ₃

XXX

H ₃ C Ν ψΌΝ

00 _ra

The title compound was prepared according to the general procedure used for the synthesis of Example 250. The compound (2.1 mg) was isolated in 10.1% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 416.16; retention time: 1.19 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 416.14; retention time: 2.04 min.

Example 254 naphthyridine-3-carbonitrile

The title compound was prepared according to the general procedure used

- 166 044500 for the synthesis of the compound of example 250. The compound (0.8 mg) was isolated in 3.2% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 97.5%; measured mass: 499.22; retention time: 1.44 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 97.3%; measured weight: 499.19; retention time: 2.2 min.

Example 255 naphthyridine-3-carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 250. The compound (9.5 mg) was isolated in 19.1% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 97.3%; measured mass: 499.2; retention time: 1.93 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 98.6%; measured mass: 499.2; retention time: 1.37 min.

Example 256 naphthyridine-3-carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 250. The compound (17.2 mg) was isolated in 36.7% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 98.6%; measured mass: 469.18; retention time: 1.89 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 167 044500 3 minutes, then maintaining 100% B for 0.75 minutes; flow rate: 1 ml/min; registration:

MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass:

469.19; retention time: 1.33 min.

Example 258 -carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 257. The compound (13 mg) was isolated in 70% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 487.93; retention time: 1.19 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 487.92; retention time: 1.81 min.

Example 259 -carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 2. The compound (4.9 mg) was isolated in 20.1% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured weight: 467.99; retention time: 1.18 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured weight: 467.99; retention time: 1.71 min.

Example 260 8-{4-[2-(4-Fluorophenyl)propan-2-yl]piperazin-1-yl}-5-methyl-7-nitro-6-oxo-5,6dihydro-1,5-naphthyridine -2-carbonitrile

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The title compound was prepared following the general procedure used for the synthesis of Example 18 using Intermediate 81. The resulting compound (18.3 mg) was isolated in 71.3% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 99.4%; measured mass: 451.22; retention time: 2.23 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 99.3%; measured mass: 451.21; retention time: 1.14 min.

Example 261 4-{4-[(4-tert-Butyl-2-hydroxyphenyl)methyl]piperazin-1-yl}-6-chloro-1-methyl-2-oxo1,2-dihydro-1,5-naphthyridine -3-carbonitrile

CH ₃

X XX

CK N y CN O OH N ^H3c \jOT ^H 3 ^C bn ^sn z (261)

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (8.4 mg) was isolated in 47.4% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 466.15; retention time: 1.51 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 466.17; retention time: 2.39 min.

Example 262 3-carbonitrile

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The title compound was prepared from 1,6-dimethyl-2-oxo-4-(piperazin-1-yl)-1,2-dihydro1,5-naphthyridine-3-carbonitrile according to the general procedure used for the synthesis of the example compound 216. The compound (9.4 mg) was isolated in 37.5% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 474.14; retention time: 2.5 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 474.14; retention time: 1.59 min.

Example 263 8-{4-[(4-Bromo-2-hydroxyphenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro1,5-naphthyridine-2,7-dicarbonitrile

The title compound was prepared from 5-methyl-6-oxo-8-(piperazin-1-yl)-5,6-dihydro-1,5naphthyridine-2,7-dicarbonitrile, 2 TFA according to the general procedure used for synthesis of the compound of Example 216. The compound (6.6 mg) was isolated in 36.2% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 95.0%; measured mass: 478.91; retention time: 1.18 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 97.4%; measured mass: 478.94; retention time: 1.89 min.

Example 264 -carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (5.9 mg) was isolated in 35.3% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 440.12; retention time: 1.19 min. Analysis conditions for 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size

- 170 044500

1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B:

(95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration:

MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass:

440.12; retention time: 1.88 min.

Example 265 7-dicarbonitrile

The title compound was prepared from 5-methyl-6-oxo-8-(piperazin-1-yl)-5,6-dihydro-1,5naphthyridine-2,7-dicarbonitrile, 2 TFA according to the general procedure used for synthesis of the compound of Example 216. The compound (5 mg) was isolated in 18.5% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 97.4%; measured mass: 485.01; retention time: 1.99 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 485.01; retention time: 1.3 min.

Example 266 -carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 2. The compound (7.9 mg) was isolated in 37.6% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 487.93; retention time: 1.26 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured weight: 487.97; retention time: 2.15 min.

Example 267 -carbonitrile

- 171 044500

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (11.7 mg) was isolated in 69.3% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile-water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 443.96; retention time: 1.77 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile-water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 443.96; retention time: 1.18 min.

Example 268 -carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (13.8 mg) was isolated in 74.3% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile-water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 99.2%; measured mass: 488.04; retention time: 2.49 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 98.1%; measured weight: 487.98; retention time: 1.5 min.

Example 269 bonitrile

- 172 044500

The title compound was prepared from 5-methyl-6-oxo-8-(piperazin-1-yl)-5,6-dihydro-1,5naphthyridine-2,7-dicarbonitrile, 2 TFA according to the general procedure used for synthesis of the compound of Example 216. The compound (6.5 mg) was isolated in 39.3% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 99.4%; measured mass: 434.97; retention time: 1.86 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 95.9%; measured mass: 434.96; retention time: 1.15 min.

Example 270 naphthyridine-3-carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 2. The compound (4.8 mg) was isolated in 20.7% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 96.6%; measured mass: 538.04; retention time: 2.63 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 538.03; retention time: 1.67 min.

Example 271 6-Bromo-4-{4-[(2-bromo-6-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridin-3 -carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 2. The compound (2.7 mg) was isolated in 14.5% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 531.89; retention time: 1.21 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B:

- 173 044500 (95:5) acetonitrile: water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration:

MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass:

531.93; retention time: 1.84 min.

Example 272 6-Chloro-4-{4-[(2-chlorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (5.7 mg) was isolated in 47.5% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 428.07; retention time: 2.19 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured weight: 428.05; retention time: 1.25 min.

Example 273 -carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (8.1 mg) was isolated in 68.2% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 424; retention time: 1.71 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured weight: 423.99; retention time: 1.16 min.

Example 274 -carbonitrile

- 174 044500

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (9.7 mg) was isolated in 41.4% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 442.12; retention time: 1.29 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 442.12; retention time: 2.04 min.

Example 275 -carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (14.7 mg) was isolated in 79.1% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 488.02; retention time: 1.52 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 99.3%; measured weight: 487.99; retention time: 2.51 min.

Example 276 -carbonitrile

- 175 044500

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (6.3 mg) was isolated in 46% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 488; retention time: 1.28 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 488.02; retention time: 2.17 min.

Example 277 4-{4-[(5-Bromo-2-hydroxyphenyl)methyl]piperazin-1-yl}-1,6-dimethyl-2-oxo-1,2dihydro-1,5-naphthyridine-3-carbonitrile

CH ₃

H ₃ C N (277)

The title compound was prepared from 1,6-dimethyl-2-oxo-4-(piperazin-1-yl)-1,2-dihydro1,5-naphthyridine-3-carbonitrile according to the general procedure used for the synthesis of the example compound 216. The compound (14.4 mg) was isolated in 78.8% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 468; retention time: 1.2 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 468.02; retention time: 2.01 min.

Example 278 -carbonitrile

CH ₃

OH N

The title compound was prepared according to the general procedure used for the synthesis of Example 2. The compound (5.6 mg) was isolated in 26.6% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured weight: 488.05; retention time: 2.48 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5)ace

- 176 044500 tonitrile: water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 99.3%; measured mass: 488.01;

retention time: 1.48 min.

Example 279 3 - carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (6 mg) was isolated in 35.7% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 442.09; retention time: 1.91 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration:

MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass:

442.13; retention time: 1.22 min.

Example 280 -carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (6.2 mg) was isolated in 38.1% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 428.01; retention time: 1.11 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 428.01; retention time: 1.86 min.

Example 281 6-Chloro-4-{4-[(2-fluoro-3-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile

- 177 044500

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (10.8 mg) was isolated in 47.8% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured weight: 425.97; retention time: 1.25 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured weight: 425.97; retention time: 2.02 min.

Example 282 -carbonitrile

CH ₃

X χ t

Br^N^Y^CN χτ (282)

The title compound was prepared according to the general procedure used for the synthesis of Example 2. The compound (7.4 mg) was isolated in 32.3% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 531.96; retention time: 2.6 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 531.98; retention time: 1.55 min.

Example 283 -carbonitrile

- 178 044500

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (7.5 mg) was isolated in 46.6% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 424.11; retention time: 2.03 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 424.11; retention time: 1.21 min.

Example 284. tert-Butyl N-(2-{ [4-(6-bromo-3-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4yl)piperazine-1 - yl]methyl}phenyl)carbamate

The title compound was prepared according to the general procedure used for the synthesis of Example 2. The compound (8.1 mg) was isolated in 34% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 553.06; retention time: 2.48 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 553.07; retention time: 1.43 min.

Example 285 carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (9.8 mg) was isolated in 38.7% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 478.13; retention time: 2.28 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) ace- 179 044500 tonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration:

MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass:

478.12; retention time: 1.4 min.

Example 286 -carbonitrile

The title compound was prepared from 1,6-dimethyl-2-oxo-4-(piperazin-1-yl)-1,2-dihydro1,5-naphthyridine-3-carbonitrile according to the general procedure used for the synthesis of the example compound 216. The compound (4.3 mg) was isolated in 23.5% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 468.01; retention time: 1.97 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured weight: 467.99; retention time: 1.16 min.

Example 287 6-Chloro-4-{4-[(2-chloro-4-fluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (9.6 mg) was isolated in 40.6% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 445.92; retention time: 1.26 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 445.91; retention time: 2.17 min.

Example 288 - carbonitrile

- 180 044500

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (9.6 mg) was isolated in 77.2% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 443.96; retention time: 1.78 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured weight: 443.95; retention time: 1.18 min.

Example 289 -3-carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (5.3 mg) was isolated in 23.5% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 96.9%; measured mass: 426.11; retention time: 2.15 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 97.2%; measured mass: 426.08; retention time: 1.32 min.

Example 290 6-Chloro-4-{4-[(3-fluoro-5-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile

- 181 044500

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (4.5 mg) was isolated in 37.7% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 426; retention time: 1.28 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 426; retention time: 2.09 min.

Example 291

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (7.3 mg) was isolated in 44.7% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 430.11; retention time: 1.26 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 430.07; retention time: 2.11 min.

Example 292 -carbonitrile

- 182 044500

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (11.2 mg) was isolated in 45.2% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 424.02; retention time: 1.16 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 95.7%; measured mass: 424.01; retention time: 1.69 min.

Example 293 -carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (12.9 mg) was isolated in 76.4% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 444.09; retention time: 1.42 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 444.08; retention time: 2.46 min.

Example 294 -carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 2. The compound (10.5 mg) was isolated in 63.5% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 471.93; retention time: 1.13 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B:

- 183 044500 (95:5) acetonitrile: water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration:

MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass:

471.92; retention time: 1.9 min.

Example 295 -carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 2. The compound (12.5 mg) was isolated in 60% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 98.3%; measured mass: 483.97; retention time: 1.81 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 98.1%; measured mass: 483.96; retention time: 1.14 min.

Example 296 -carbonitrile

The title compound was prepared from 1,6-dimethyl-2-oxo-4-(piperazin-1-yl)-1,2-dihydro1,5-naphthyridine-3-carbonitrile according to the general procedure used for the synthesis of the example compound 216. The compound (10.5 mg) was isolated in 46.7% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 424.09; retention time: 2.4 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 424.11; retention time: 1.43 min.

Example 297 6-Bromo-4-{4-[(2-methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro1,5-naphthyridine-3-carbonitrile

- 184 044500

The title compound was prepared according to the general procedure used for the synthesis of Example 2. The compound (5.2 mg) was isolated in 31.7% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 95.8%; measured mass: 468; retention time: 1.22 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 95.2%; measured mass: 468; retention time: 1.8 min.

Example 298 -3-carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (11.6 mg) was isolated in 68.5% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 96.6%; measured mass: 446.07; retention time: 2.32 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 98.9%; measured mass: 446.1; retention time: 1.4 min.

Example 299 1,5-naphthyridine-3-carbonitrile

- 185 044500

The title compound was prepared according to the general procedure used for the synthesis of Example 2. The compound (5.4 mg) was isolated in 26.2% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 478.98; retention time: 1.19 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured weight: 478.99; retention time: 1.94 min.

Example 3 00. 6-Bromo-4-{4-[(2-chlorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 2. The compound (7.3 mg) was isolated in 44.1% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 472; retention time: 2.23 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured weight: 471.99; retention time: 1.3 min.

Example 301 -carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (6.9 mg) was isolated in 56% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 440.01; retention time: 1.58 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 186 044500 3 minutes, then maintaining 100% B for 0.75 minutes; flow rate: 1 ml/min; registration:

MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass:

439.98; retention time: 1.13 min.

Example 302 6-Chloro-4-{4-[(3-chlorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (14.3 mg) was isolated in 69.6% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured weight: 428.05; retention time: 1.3 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 428.07; retention time: 2.17 min.

Example 303 3-carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (6.3 mg) was isolated in 38.3% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 433.02; retention time: 2.03 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured weight: 432.99; retention time: 1.25 min.

Example 304 6-Bromo-4-{4-[(3-fluoro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile

- 187 044500

The title compound was prepared according to the general procedure used for the synthesis of Example 2. The compound (6.6 mg) was isolated in 32.5% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 97.2%; measured mass: 472.04; retention time: 2.33 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 472.06; retention time: 1.37 min.

Example 305 -carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (8.7 mg) was isolated in 52% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 440.13; retention time: 1.13 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 440.12; retention time: 1.8 min.

Example 306 -carbonitrile

The title compound was prepared according to the general procedure used

- 188 044500 for the synthesis of the compound of example 216. The compound (11.6 mg) was isolated in 71.3% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured weight: 428.09; retention time: 1.32 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 428.1; retention time: 2.28 min.

Example 307 6-Chloro-4-{4-[(3,5-dichloro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo1,2-dihydro-1,5-naphthyridine -3-carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (7.9 mg) was isolated in 43.4% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 478.02; retention time: 1.65 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 478.03; retention time: 2.66 min.

Example 308

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (12.2 mg) was isolated in 61.7% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 412.1; retention time: 1.97 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) ace- 189 044500 tonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration:

MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 91.9%; measured weight: 412.07;

retention time: 1.21 min.

Example 309 6-Bromo-4-{4-[(3-tert-butyl-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo1,2-dihydro-1,5-naphthyridine -3-carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 2. The compound (3.6 mg) was isolated in 16.4% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 510.09; retention time: 3.04 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 510.12; retention time: 1.96 min.

Example 310

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (3.1 mg) was isolated in 12.6% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 462.03; retention time: 2.46 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 462.07; retention time: 1.43 min.

Example 311 naphthyridine-3-carbonitrile

- 190 044500

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (4.8 mg) was isolated in 26.4% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 478.07; retention time: 2.18 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 478.11; retention time: 1.34 min.

Example 312 6-Chloro-4-{4-[(5-fluoro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridin-3 -carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (8.6 mg) was isolated in 52.9% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 428.12; retention time: 1.92 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 428.1; retention time: 1.14 min.

Example 313 6-Chloro-4-(4-{[3-fluoro-5-(trifluoromethyl)phenyl]methyl}piperazin-1-yl)-1-methyl-2-oxo1,2-dihydro-1,5- naphthyridine-3-carbonitrile

- 191 044500

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (7.4 mg) was isolated in 40.6% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 480.09; retention time: 1.44 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 480.09; retention time: 2.3 min.

Example 314 -carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 2. The compound (7.4 mg) was isolated in 43.7% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured weight: 483.98; retention time: 1.17 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured weight: 483.98; retention time: 1.79 min.

Examples 315 and 316 6-Chloro-4-{4-[1-(4-fluorophenyl)-2-methylpropyl]piperazin-1-yl}-1-methyl-2oxo-1,2-dihydro-1,5- naphthyridine-3-carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 193. This racemic compound (17.5 mg) was isolated in 55.9% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 454.17; retention time: 2.46 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid;

- 192 044500 mobile phase B: (95:5) acetonitrile: water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for 2nd sample injection: purity:

100.0%; measured mass: 454.14; retention time: 1.39 min. The racemic mixture was separated to give Example 315 (isomer 1) and Example 316 (isomer 2).

Example 315 (4.3 mg) was isolated in 27.1% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 454.42; retention time: 2.61 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 454.15; retention time: 1.34 min.

Example 316 (4.2 mg) was isolated in 26.4% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 454.1; retention time: 2.62 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 99.4%; measured mass: 454.26; retention time: 1.35 min.

Example 317 8-{4-[(2-Hydroxyphenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2,7-dicarbonitrile

The title compound was prepared from 5-methyl-6-oxo-8-(piperazin-1-yl)-5,6-dihydro-1,5naphthyridine-2,7-dicarbonitrile, 2 TFA according to the general procedure used for synthesis of the compound of Example 216. The compound (6 mg) was isolated in 39.4% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 99.1%; measured mass: 400.98; retention time: 1.02 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 98.8%; measured weight: 400.99; retention time: 1.62 min.

Example 318

- 193 044500

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (13.2 mg) was isolated in 64% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured weight: 430.05; retention time: 2 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 430.08; retention time: 1.2 min.

Example 319 6-Chloro-4-{4-[(3,4-difluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro1,5-naphthyridine-3-carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (3.9 mg) was isolated in 32.4% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 430.07; retention time: 1.25 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 430.09; retention time: 2.07 min.

Example 320 6-Bromo-4-{4-[(3,5-difluoro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo1,2-dihydro-1,5-naphthyridine -3-carbonitrile

- 194 044500

The title compound was prepared according to the general procedure used for the synthesis of Example 2. The compound (9.8 mg) was isolated in 46.5% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 98.9%; measured mass: 490.02; retention time: 2.39 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 98.9%; measured weight: 489.98; retention time: 1.42 min.

Example 321

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (4.5 mg) was isolated in 39% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured weight: 412.07; retention time: 1.2 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 412.13; retention time: 2.02 min.

Example 322 3-carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (3 mg) was isolated in 24.7% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 434.11; retention time: 1.8 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) ace- 195 044500 tonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration:

MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass:

434.07; retention time: 1.11 min.

Example 323 -carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (5 mg) was isolated in 40.2% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 444.08; retention time: 1.77 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 444.06; retention time: 1.19 min.

Example 324 -3-carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (4.3 mg) was isolated in 25.4% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured weight: 446.05; retention time: 2.27 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 446.04; retention time: 1.34 min.

Example 325 8-[4-(Diphenylmethyl)piperazin-1-yl]-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2,7dicarbonitrile

- 196 044500

Zinc (0.696 mg, 10.64 µmol), bromo(tri-tertbutylphosphine)palladium(I) dimer (8.27 mg, 10.64 µmol), dicyanozinc (1.999 mg, 0.017 mmol) and 4 -(4benzhydrylpiperazin-1 -yl)-6-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3-carbonitrile (10 mg, 0.021 mmol). The tube was hermetically sealed, placed under vacuum and filled with nitrogen. DMF (0.5 ml) was added and the reaction mixture was heated for 2 hours at 50°C. According to LC-MS analysis, the reaction was complete. The reaction mixture was diluted by adding CH3CN, filtered and the filtrate was purified by preparative LC-MS using the following column conditions:

XBridge C18, 19x200 mm, particle size 5 microns; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 5090% B for 15 min, then maintain 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. The compound (4 mg) was isolated in 41.4% yield. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Example 326 6-Chloro-4-{4-[(4-chloro-3-fluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (5.6 mg) was isolated in 33% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 446.09; retention time: 1.36 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 446.08; retention time: 2.22 min.

Example 327 6-Chloro-4-{4-[(3-methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro1,5-naphthyridine-3-carbonitrile

- 197 044500

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (5.3 mg) was isolated in 31.3% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 424.1; retention time: 1.91 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 424.09; retention time: 1.24 min.

Example 328 -carbonitrile

The title compound was prepared according to the general procedure used for the synthesis of Example 216. The compound (6.3 mg) was isolated in 53.1% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 424.1; retention time: 1.19 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 424.08; retention time: 1.72 min.

Example 329 -carbonitrile

The title compound was prepared from 1,6-dimethyl-2-oxo-4-(piperazin-1-yl)-1,2-dihydro

- 198 044500

1,5-naphthyridine-3-carbonitrile according to the general procedure used for the synthesis of Example 216. The compound (9.6 mg) was isolated in 44.5% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 408.16; retention time: 1.27 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 408.16; retention time: 2.11 min.

Example 330 6-Chloro-4-{4-[(2,3-difluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro1,5-naphthyridine-3-carbonitrile

Example 330 was prepared following the general procedure used for the synthesis of Example 216. The compound (6.1 mg) was isolated in 26.8% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 99.1%; measured weight: 429.94; retention time: 1.94 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 97.3%; measured weight: 429.94; retention time: 1.19 min.

Example 331 6-Chloro-4-{4-[(2-methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro1,5-naphthyridine-3-carbonitrile

Example 331 was prepared following the general procedure used for the synthesis of Example 216. The compound (2.9 mg) was isolated in 18% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 424.03; retention time: 1.2 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonite-199 044500 ril:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 424.02; retention time: 1.76 min.

Example 332 8-{4-[(4-Chloro-3-hydroxyphenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro1,5-naphthyridine-2,7-dicarbonitrile

Example 332 was prepared from 5-methyl-6-oxo-8-(piperazin-1-yl)-5,6-dihydro-1,5naphthyridine-2,7-dicarbonitrile, 2 TFA according to the general procedure used for the synthesis compound of example 216. The compound (14 mg) was isolated in 84.7% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 98.9%; measured mass: 434.94; retention time: 1.54 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 98.8%; measured mass: 434.96; retention time: 1.1 min.

Example 333 3-carbonitrile

The compound of Example 333 was prepared according to the general procedure used for the synthesis of the compound of Example 2. The compound (3.5 mg) was isolated in 20.9% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 98.5%; measured mass: 478.04; retention time: 1.14 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 98.6%; measured mass: 478.08; retention time: 1.84 min.

Example 334 -carbonitrile

- 200 044500

The compound of Example 334 was prepared according to the general procedure used for the synthesis of the compound of Example 66. The compound (21.3 mg) was isolated in 73.4% yield. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Example 335 -carbonitrile

Example 335 was prepared following the general procedure used for the synthesis of Example 216. The compound (8.7 mg) was isolated in 37.1% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 442.13; retention time: 1.89 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 442.1; retention time: 1.25 min.

Example 336 -carbonitrile

The compound of Example 336 was prepared according to the general procedure used for the synthesis of the compound of Example 216. The compound (10.4 mg) was isolated in 19.2% yield. To determine the final

- 201 044500 product purity, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Example 337

Example 337 was prepared from 1,6-dimethyl-2-oxo-4-(piperazin-1-yl)-1,2-dihydro-1,5naphthyridine-3-carbonitrile according to the general procedure used for the synthesis of Example 216 The compound (5.9 mg) was isolated in 28.6% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 390.16; retention time: 2.13 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 99.3%; measured mass: 390.15; retention time: 1.27 min.

Example 338 -carbonitrile

Example 338 was prepared following the general procedure used for the synthesis of Example 216. The compound (4.9 mg) was isolated in 40.9% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 97.9%; measured mass: 428.07; retention time: 1.13 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV

- 202 044500 (220 nm). Analysis results for the 2nd sample injection: purity: 98.2%; measured weight: 428.07; retention time: 1.68 min.

Example 339 6-Chloro-4-{4-[(2-fluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile

Example 339 was prepared following the general procedure used for the synthesis of Example 216. The compound (5.9 mg) was isolated in 51.2% yield. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Example 340 6-Chloro-4-{4-[(5-cyano-2-fluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile

Example 340 was prepared following the general procedure used for the synthesis of Example 216. The compound (7.4 mg) was isolated in 32% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 99.2%; measured mass: 437.09; retention time: 1.13 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 99.2%; measured mass: 437.07; retention time: 1.83 min.

Example 341 6-Chloro-4-{4-[(4-methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro1,5-naphthyridine-3-carbonitrile

- 203 044500

The compound of Example 341 was prepared according to the general procedure used for the synthesis of the compound of Example 216. The compound (5.2 mg) was isolated in 30.7% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 424.1; retention time: 1.86 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 424.11; retention time: 1.22 min.

Example 342

Example 342 was prepared following the general procedure used for the synthesis of Example 221. The compound (14.8 mg) was isolated in 57.9% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 449.23; retention time: 1.27 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 99.4%; measured mass: 449.1; retention time: 2.14 min.

Example 343 4-(4-Benzylpiperazin-1-yl)-6-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3carbonitrile

Example 343 was prepared according to the general procedure used for the synthesis

- 204 044500 compound of example 216. The compound (6.9 mg) was isolated in 46.1% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 394.1; retention time: 1.19 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 394.1; retention time: 1.95 min.

Example 344

CH ₃ (344)

The compound of Example 344 was prepared according to the general procedure used for the synthesis of the compound of Example 2. The compound (8.6 mg) was isolated in 53.8% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 455.96; retention time: 1.91 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 455.96; retention time: 1.17 min.

Example 345 - carbonitrile

CH ₃

Example 345 was prepared following the general procedure used for the synthesis of Example 216. The compound (4.4 mg) was isolated in 35.4% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 99.3%; measured mass: 444.09; retention time: 1.82 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) ace- 205 044500 tonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration:

MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 99.0%; measured mass: 444.1;

retention time: 1.22 min.

Example 346

Example 346 was prepared following the general procedure used for the synthesis of Example 216. The compound (7.4 mg) was isolated in 64.8% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 408.11; retention time: 1.26 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 408.12; retention time: 2.22 min.

Example 347 4-{4-[bis(4-Fluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile

Example 347 was prepared following the general procedure used for the synthesis of Example 66. The compound (23.1 mg) was isolated in 72% yield. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Example 348 -carbonitrile

- 206 044500

Example 348 was prepared following the general procedure used for the synthesis of Example 216. The compound (6.7 mg) was isolated in 41.2% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 428.08; retention time: 1.09 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 428.1; retention time: 1.62 min.

Example 349

The compound of Example 349 was prepared according to the general procedure used for the synthesis of the compound of Example 66. The compound (22.2 mg) was isolated in 72.3% yield. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1 x 50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1 x 50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Example 350

Example 350 was prepared following the general procedure used for the synthesis of Example 216. The compound (5.9 mg) was isolated in 34.3% yield. To determine the final

- 207 044500 analytical LC-MS was used for product purity. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 430.07; retention time: 2 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 430.06; retention time: 1.23 min.

Example 351 6-Chloro-4-{4-[(3-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro1,5-naphthyridine-3-carbonitrile

Example 351 was prepared following the general procedure used for the synthesis of Example 216. The compound (4.5 mg) was isolated in 39.2% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 410.11; retention time: 1.56 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured weight: 410.06; retention time: 1.06 min.

Example 352

Example 352 was prepared from 1,6-dimethyl-2-oxo-4-(piperazin-1-yl)-1,2-dihydro-1,5naphthyridine-3-carbonitrile according to the general procedure used for the synthesis of Example 216 The compound (6.6 mg) was isolated in 29.7% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for 1st sample injection: column: Waters

XBridge C18, 2.1 mmx50 mm, particle size 1.7 microns; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 420.14; retention time: 1.41 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Results - 208 044500 analysis for the 2nd sample injection: purity: 100.0%; measured weight: 420.15; retention time: 2.04 min.

Example 353 -carbonitrile

Example 353 was prepared following the general procedure used for the synthesis of Example 216. The compound (6.8 mg) was isolated in 40.7% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 95.8%; measured mass: 440.08; retention time: 1.55 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 95.9%; measured mass: 440.12; retention time: 1.08 min.

Example 354 -carbonitrile

Example 354 was prepared following the general procedure used for the synthesis of Example 216. The compound (6.5 mg) was isolated in 36.6% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 444.09; retention time: 1.66 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 444.09; retention time: 1.14 min.

Example 355 naphthyridine-3-carbonitrile

- 209 044500

Example 355 was prepared following the general procedure used for the synthesis of Example 216. The compound (5.4 mg) was isolated in 21.3% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 99.3%; measured mass: 478.11; retention time: 1.27 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 99.4%; measured mass: 478.13; retention time: 1.79 min.

Example 356 -carbonitrile

Example 356 was prepared following the general procedure used for the synthesis of Example 216. The compound (8.7 mg) was isolated in 38.7% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured weight: 423.99; retention time: 1.49 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 424; retention time: 1.09 min.

Example 357 -carbonitrile

- 210 044500

Example 357 was prepared following the general procedure used for the synthesis of Example 216. The compound (5.9 mg) was isolated in 49.2% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 428.07; retention time: 1.06 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 428.07; retention time: 1.56 min.

Example 358

Example 358 was prepared following the general procedure used for the synthesis of Example 191. The compound (4.2 mg) was isolated in 15.1% yield. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1 x 50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Example 359 ,5-naphthyridine-3-carbonitrile

CH ₃

Example 359 was prepared following the general procedure used for the synthesis of Example 216. The compound (4.5 mg) was isolated in 27.3% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 96.2%; measured mass: 434.09; retention time: 0.94 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV

- 211 044500 (220 nm). Analysis results for the 2nd sample injection: purity: 97.4%; measured mass: 434.1; retention time: 1.47 min.

Example 360 -naphthyridine-3-carbonitrile

The compound of Example 360 was prepared according to the general procedure used for the synthesis of the compound of Example 2. The compound (3.6 mg) was isolated in 21.5% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for 1st sample input: column:

Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 microns; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 478.01; retention time: 1.5 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 478.04; retention time: 0.95 min.

Example 362 6-Chloro-4-{4-[(4-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro1,5-naphthyridine-3-carbonitrile

Example 362 was prepared following the general procedure used for the synthesis of Example 216. The compound (5.7 mg) was isolated in 36.6% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 410.12; retention time: 1.46 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 410.12; retention time: 1.03 min.

Example 363

- 212 044500

Example 363 was prepared from 5-methyl-6-oxo-8-(piperazin-1-yl)-5,6-dihydro-1,5-naphthyridine-2,7-dicarbonitrile, 2 TFA according to the general procedure used to synthesize the compound of Example 216. The compound (7.4 mg) was isolated in 30.1% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 419.01; retention time: 0.95 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 419.01; retention time: 1.32 min.

Example 364 6-Chloro-4-{4-[(2-fluoro-5-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile

Example 364 was prepared following the general procedure used for the synthesis of Example 216. The compound (9.1 mg) was isolated in 40.3% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 99.4%; measured mass: 426.14; retention time: 1.29 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 426.11; retention time: 2.1 min.

Example 365 -carbonitrile

- 213 044500

Example 365 was prepared following the general procedure used for the synthesis of Example 216. The compound (10.1 mg) was isolated in 41.6% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 99.2%; measured weight: 458.09; retention time: 2.13 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 99.2%; measured mass: 458.06; retention time: 1.34 min.

Example 366 6-Bromo-4-{4-[(3,5-dichloro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo1,2-dihydro-1,5-naphthyridine -3-carbonitrile

Example 366 was prepared following the general procedure used for the synthesis of Example 2. The compound (8.3 mg) was isolated in 36.9% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 98.3%; measured mass: 521.97; retention time: 2.76 min.

Example 367 -carbonitrile

The compound of Example 367 was prepared according to the general procedure used for the synthesis of the compound of Example 2. The compound (11.1 mg) was isolated in 52.8% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 488; retention time: 2.56 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 488; retention time: 1.5 min.

Example 368 -naphthyridine-3-carbonitrile

The compound of Example 368 was prepared according to the general procedure used for the synthesis of the compound of Example 2. The compound (14 mg) was isolated in 67.2% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for 1st sample input: column:

Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 microns; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 97.6%; measured mass: 484.05; retention time: 1.41 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 97.2%; measured mass: 484.09; retention time: 2.18 min.

Example 369 naphthyridine-3-carbonitrile

Example 369 was prepared following the general procedure used for the synthesis of Example 216. The compound (12.5 mg) was isolated in 66.6% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 494.05; retention time: 1.64 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 97.0%; measured mass: 494.08; retention time: 2.6 min.

Example 370 -carbonitrile

- 215 044500

Example 370 was prepared following the general procedure used for the synthesis of Example 216. The compound (11.7 mg) was isolated in 69.3% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 444.1; retention time: 2.5 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 im). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 444.07; retention time: 1.48 min.

Example 371 -carbonitrile

Example 371 was prepared following the general procedure used for the synthesis of Example 216. The compound (9.4 mg) was isolated in 56.2% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 440.11; retention time: 1.37 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 440.11; retention time: 2.13 min.

Example 372 6-Bromo-4-{4-[(2-hydroxy-4-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridin-3 -carbonitrile

The compound of Example 372 was prepared according to the general procedure used for the synthesis of the compound of Example 2. The compound (10.1 mg) was isolated in 50.2% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 468.07; retention time: 1.22 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonite-216 044500 ril:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured weight: 468.05; retention time: 2.07 min.

Example 373 3-carbonitrile

The compound of Example 373 was prepared according to the general procedure used for the synthesis of the compound of Example 2. The compound (11.3 mg) was isolated in 55.1% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for 1st sample input: column:

Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 microns; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 98.8%; measured mass: 477.08; retention time: 2.13 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 477.08; retention time: 1.31 min.

Example 374 ,2-dihydro-1,5-naphthyridine-3-carbonitrile

The compound of Example 374 was prepared according to the general procedure used for the synthesis of the compound of Example 2. The compound (5.8 mg) was isolated in 27.3% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 493.37; retention time: 0.91 min.

Example 375 6-chloro-4-(4-{[3-fluoro-4-(trifluoromethyl)phenyl]methyl}piperazin-1-yl)-1-methyl-2-oxo1,2-dihydro-1,5- naphthyridine-3-carbonitrile

- 217 044500

Example 375 was prepared following the general procedure used for the synthesis of Example 216. The compound (8.1 mg) was isolated in 44.4% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 480.1; retention time: 1.47 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 480.1; retention time: 2.27 min.

Example 376 6-Chloro-1-methyl-2-oxo-4-{4-[(2-oxo-2,3-dihydro-1H-indol-7-yl)methyl]piperazin-1yl}-1,2 -dihydro-1,5-naphthyridine-3-carbonitrile

The compound of Example 376 was prepared according to the general procedure used for the synthesis of the compound of Example 216. The compound (4 mg) was isolated in 31.8% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured weight: 448.98; retention time: 1.44 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 449; retention time: 1.07 min.

Examples 377 and 378 5-naphthyridine-3-carbonitrile

Compounds of Examples 377-378 were prepared according to the general procedure used for the synthesis of Example 68. This racemic compound (25 mg) was isolated in 60.3% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 99.1%; measured mass: 482.32; retention time: 2.52 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from

0% B to 100% B for 3 min, then maintain 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 99.4%;

measured mass: 482.06; retention time: 1.69 min.

The racemic material was further purified using SFC-chiral chromatography to obtain the compound of Example 377 (first eluting isomer) and the compound of Example 378 (second eluting isomer).

Example 377 The compound (9.4 mg) was isolated in 22.7% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 99.0%; measured mass: 482.31; retention time: 2.51 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 482.36; retention time: 1.56 min.

Example 378 The compound (9.5 mg) was isolated in 22.9% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 98.1%; measured mass: 482.32; retention time: 2.51 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 99.3%; measured mass: 482.31; retention time: 1.56 min.

Example 379 -carbonitrile

Example 379 was prepared following the general procedure used for the synthesis of Example 216. The compound (1.8 mg) was isolated in 7.8% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 99.1%; measured mass: 436.92; retention time: 1.13 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 98.7%; measured mass: 436.93; retention time: 1.8 min.

Example 380 -carbonitrile

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The compound of Example 380 was prepared according to the general procedure used for the synthesis of the compound of Example 2. The compound (2.3 mg) was isolated in 10% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 98.2%; measured mass: 531.93; retention time: 1.52 min.

Example 381 naphthyridine-3-carbonitrile

Example 381 was prepared following the general procedure used for the synthesis of Example 216. The compound (4.3 mg) was isolated in 22.9% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 494.1; retention time: 2.23 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 494.09; retention time: 1.4 min.

Examples 382A, 382 and 383 3-carbonitrile

CH ₃

C.I.

(382A, 382-383)

Example 382A was prepared following the general procedure used for the synthesis of Example 68. The compound (24.6 mg) was isolated in 63.6% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for 1

- 220 044500 th sample input: purity: 100.0%; measured mass: 466.12; retention time: 2.39 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 466.08; retention time: 1.38 min. The racemic material was separated using SFC-chiral chromatography to obtain Example 382 (first eluting isomer) and Example 383 (second eluting isomer).

Example 382 The compound (7.7 mg) was isolated in 19.9% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 95.5%; measured mass: 466.06; retention time: 2.63 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 95.4%; measured mass: 466.08; retention time: 1.55 min.

Example 383 The compound (7.7 mg) was isolated in 19.9% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 96.7%; measured mass: 466.04; retention time: 2.63 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 96.2%; measured mass: 466.08; retention time: 1.55 min.

Example 384 -3-carbonitrile

Example 384 was prepared following the general procedure used for the synthesis of Example 216. The compound (1.3 mg) was isolated in 7.3% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 466.17; retention time: 1.86 min.

Example 385

- 221 044500

Example 385 was prepared following the general procedure used for the synthesis of Example 216. The compound (9.5 mg) was isolated in 48.5% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 408.12; retention time: 1.3 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 408.13; retention time: 2.08 min.

Example 3 86. 6-Chloro-1-methyl-4-{4-[(4-methylphenyl)methyl]piperazin-1 -yl}-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile

Example 386 was prepared following the general procedure used for the synthesis of Example 216. The compound (18.8 mg) was isolated in 29% yield. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1 x 50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm.

Examples 387A, 387 and 388 -3-carbonitrile

Example 387A was prepared following the general procedure used for the synthesis of Example 68. The compound (16.5 mg) was isolated in 56.1% yield. To determine the final

- 222 044500 analytical LC-MS was used for product purity. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured weight: 425.97; retention time: 1.37 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 im). Analysis results for the 2nd sample injection: purity: 100.0%; measured weight: 425.98; retention time: 2 min. The racemic material was further purified using SFC chiral chromatography to provide Example 387 (first eluting isomer) and Example 388 (second eluting isomer).

Example 387 The compound (4.4 mg) was isolated in 15.9% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 98.4%; measured mass: 426.18; retention time: 2.05 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 98.7%; measured mass: 426.21; retention time: 1.15 min.

Example 388 Compound (1.7 mg) was isolated in 6.1% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 99.1%; measured mass: 426.16; retention time: 2.05 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 98.8%; measured mass: 426.2; retention time: 1.15 min.

Example 389 -carbonitrile

Example 389 was prepared from 1,6-dimethyl-2-oxo-4-(piperazin-1-yl)-1,2-dihydro-1,5naphthyridine-3-carbonitrile according to the general procedure used for the synthesis of Example 216 The compound (4.4 mg) was isolated in 10.8% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 424.14; retention time: 1.51 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase

- 223 044500

A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 424.11; retention time: 2.39 min.

Example 390 -carbonitrile

The compound of Example 390 was prepared according to the general procedure used for the synthesis of the compound of Example 2. The compound (0.8 mg) was isolated in 4.7% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 488; retention time: 1.25 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 488; retention time: 2.14 min.

Example 391 -carbonitrile

Example 391 was prepared following the general procedure used for the synthesis of Example 216. The compound (5.6 mg) was isolated in 34.8% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 424.08; retention time: 2.12 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 424.08; retention time: 1.23 min.

Example 392 2-carbonitrile

- 224 044500

8-Chloro-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (19.8 mg, 0.090 mmol) was suspended in DMF (0.9 ml) in a 1 dram tube . To this mixture was added 1-((4-fluoro-2methoxyphenyl)(4-fluorophenyl)methyl)piperazine, 2 HCl (42.3 mg, 0.108 mmol) and potassium carbonate (62.3 mg, 0.451 mmol). The reaction mixture was hermetically sealed under a nitrogen atmosphere and immersed in an oil bath at 120°C for 6 hours. LC-MS analysis showed the formation of a new peak corresponding to the molecular weight of the desired product. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 200 mmx19 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: maintain 27% B for 0 min, 2777% B for 25 min, then maintain 100% B for 4 min; flow rate: 20 ml/min; column temperature: 25 C. Fraction collection was triggered by MS and UV signals. Fractions containing the desired product were combined and dried using a centrifugal evaporator. The compound (12.7 mg) was isolated in 28.1% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 502.19; retention time: 1.47 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 502.19; retention time: 2.27 min. 1H NMR (500 MHz, DMSO-d ₆ ) δ 8.21-8.12 (m, 1.0H), 8.10-8.03 (m, 1.0H), 7.60 (b.t, J=7.6 Hz, 1.0H), 7.45-7.38 (m, 2.1Н), 7.13 (br.t, J=8.7 Hz, 2.1Н), 6.87 (br.d, J=11.3 Hz, 1.0Н), 6.81 (br.t, J=8.2 Hz, 1.1Н ), 6.09 (s, 1.0H), 4.76 (s, 1.0H), 3.80 (s, 2.9H), 3.53 (s, 1.9H), 3.47 (br. s, 0.9H), 2.60-2.53 (m, 2.0H), 2.47-2.39 (m, 2.0H).

Example 393

The title compound was prepared according to the general procedure used for the synthesis of Example 191. The compound (35.8 mg) was isolated in 61.4% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 582.01; retention time: 2.01 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass:

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581.98; retention time: 2.72 min.

Example 394

The compound of Example 394 was prepared according to the general procedure used for the synthesis of the compound of Example 325. The compound (8.4 mg) was isolated in 58.8% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 529.13; retention time: 2.52 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 97.9%; measured mass: 529.11; retention time: 1.89 min.

Example 395 carbonitrile

4-(4-(bis(4-chlorophenyl)methyl)piperazin-1-yl)-6bromo-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3 was added to a microwave test tube -carbonitrile (15 mg, 0.026 mmol), palladium(II) acetate (0.577 mg, 2.57 µmol), cesium carbonate (8.38 mg, 0.026 mmol) and 5-[di(1-adamantyl)phosphino]-1 ',3',5'triphenyl-1'h-[1,4']bipyrazole (3.41 mg, 5.14 µmol). The tube was placed under vacuum, washed several times with nitrogen and hermetically sealed. Methanol (0.1 ml) and acetonitrile (2 ml) were added and the reaction mixture was heated at 80°C overnight. According to LC-MS analysis, the reaction was complete. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 200 mmx19 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; gradient: maintaining 28% B for 0 min, 28-68% B for 20 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min; column temperature: 25 C. Fraction collection was triggered by MS and UV signals. Fractions containing the desired product were combined and dried using a centrifugal evaporator. The title compound (13.5 mg) was isolated in 97.2% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 534.17; retention time: 2.59 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0% B to 100% B for 3 min, then maintain 100% B for 0.75

- 226 044500 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 534.17; retention time: 1.87 min.

Example 396 5-naphthyridine-3-carbonitrile

6-Methoxy-1-methyl-2-oxo-4-(piperazin-1-yl)-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (30 mg, 0.100 mmol), (4-fluorophenyl) (2-methoxypyridin-3-yl)methanol (28.1 mg, 0.120 mmol), (cyanomethyl)trimethylphosphonium iodide (48.7 mg, 0.200 mmol) were mixed with propionitrile (0.8 ml) in a hermetically sealed tube of 2 ml. Huenig's base (0.053 mL, 0.301 mmol) was added and the reaction mixture was sealed and heated at 110° C. in a microwave reactor for 4 hours. The reaction was complete by LC-MS analysis. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 200 mmx19 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: maintaining 32% B for 0 min, 32-72% B for 25 min, then maintaining 100% B for 4 min; flow rate: 20 ml/min; Column temperature: 25°C. Fraction collection was triggered by MS and UV signals. Fractions containing the desired product were combined and dried using a centrifugal evaporator. The compound (11.4 mg) was isolated in 22.2% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 98.2%; measured mass: 515.2; retention time: 2.14 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 97.2%; measured mass: 515.22; retention time: 1.41 min.

Example 397 5-naphthyridine-3-carbonitrile

Example 397 was prepared following the general procedure used for the synthesis of Example 396. The compound (4.8 mg) was isolated in 9.3% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 99.1%; measured mass: 515.13; retention time: 1.86 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm).

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Analysis results for the 2nd sample injection: purity: 98.9%; measured mass: 515.13; retention time: 1.36 min.

Examples 398, 398A and 398B. 4-{4-[1-(4-Fluorophenyl)propyl]piperazin-1-yl}-1,6-dimethyl-2-oxo-1,2dihydro-1,5-naphthyridine-3-carbonitrile

Example 398 was prepared following the general procedure used for the synthesis of Example 226. The compound (28.8 mg) was isolated in 64.2% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 420.18; retention time: 2.08 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 97.9%; measured mass: 420.18; retention time: 1.37 min. The racemic material was further purified using SFC chiral chromatography to provide Example 398A (first eluting isomer) and Example 398B (second eluting isomer).

Example 398A (9.5 mg) was isolated in 33.8% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 420.11; retention time: 2.21 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 420.31; retention time: 1.23 min.

Example 398B (10.2 mg) was isolated in 36.3% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 420.1; retention time: 2.21 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 420.29; retention time: 1.23 min.

Example 399 naphthyridine-2-carbonitrile

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Example 399 was prepared following the general procedure used for the synthesis of Example 18. The compound (8.5 mg) was isolated in 43.4% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 515.14; retention time: 2.5 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 515.17; retention time: 1.72 min.

Example 400 2,7-dicarbonitrile

The compound of Example 400 was prepared from 5-methyl-6-oxo-8-(piperazin-1-yl)-5,6-dihydro-1,5 naphthyridine-2,7-dicarbonitrile according to the general procedure used for the synthesis of the compound of Example 396. The compound (5.5 mg) was isolated in 23.4% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 96.9%; measured mass: 480.16; retention time: 1.81 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 im). Analysis results for the 2nd sample injection: purity: 99.1%; measured mass: 480.16; retention time: 1.29 min.

Example 401 1,5-naphthyridine-3-carbonitrile

- 229 044500

Example 401 was prepared following the general procedure used for the synthesis of Example 396. The compound (12.6 mg) was isolated in 24.4% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 96.1%; measured mass: 516.21; retention time: 1.64 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 96.8%; measured mass: 516.2; retention time: 1.22 min.

Example 402 3-carbonitrile

Example 402 was prepared following the general procedure used for the synthesis of Example 396. The compound (33.9 mg) was isolated in 60.4% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 562.2; retention time: 1.56 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 562.19; retention time: 2.31 min.

Example 403 naphthyridine-3-carbonitrile

Example 403 was prepared following the general procedure used for the synthesis of Example 396. The compound (10.8 mg) was isolated in 22.3% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 98.6%; measured mass: 485.17; retention time: 1.85 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) ace- 230 044500 tonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration:

MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 98.9%; measured mass: 485.18;

retention time: 1.32 min.

Example 404 5-Methyl-8-{4-[(naphthalene-1 -yl)methyl]piperazin-1 -yl}-6-oxo-5,6-dihydro-1,5-naphthyridine-2,7-dicarbonitrile

Example 404 was prepared from 6-bromo-1-methyl-4-(4-(naphthalene-1-ylmethyl)piperazin-1-yl)2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile following the general procedure used for the synthesis of Example 52. The compound (3.7 mg) was isolated in 8.3% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 90.2%; measured mass: 435.14; retention time: 1.27 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 90.1%; measured mass: 435.16; retention time: 2.07 min.

Examples 405-407. 8-{4-[(4-Fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl]piperazin-1-yl}-5-methyl-6oxo-5,6-dihydro-1,5-naphthyridine-2,7 -dicarbonitrile

The compound of Examples 405-407 was prepared from 6-bromo-4-(4-((4-fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl)piperazin-1-yl)-1-methyl-2-oxo-1, 2-dihydro-1,5-naphthyridine-3-carbonitrile according to the general procedure used for the synthesis of Example 52. This compound (12.1 mg) was isolated in 33.3% yield. Analytical LCMS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 97.8%; measured mass: 527.16; retention time: 1.65 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 96.1%; measured mass: 527.17; retention time: 2.24 min. The racemic material was further purified using SFC-chiral chromatography to obtain the compound of Example 406 (first eluting isomer) and the compound of Example 407 (second eluting isomer).

Example 406 The title compound (3.5 mg) was isolated in 31.7% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for 1st input

- 231 044500 sample: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 95.2%; measured mass: 527.08; retention time: 2.15 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 97.9%; measured mass: 526.97; retention time: 2.06 min.

Example 407 The title compound (27.8 mg) was isolated in 61.4% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 527.14; retention time: 1.5 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 527.14; retention time: 2.25 min.

Example 408 5-Methyl-8-{4-[(4-methylphenyl)(phenyl)methyl]piperazin-1-yl}-6-oxo-5,6-dihydro-1,5naphthyridine-2,7-dicarbonitrile

The compound of Example 408 was prepared according to the general procedure used for the synthesis of the compound of Example 52. The compound (2.6 mg) was isolated in 45.7% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 475.25; retention time: 1.52 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 98.1%; measured mass: 475.24; retention time: 2.32 min.

Example 409 7-dicarbonitrile

- 232 044500

Example 409 was prepared following the general procedure used for the synthesis of Example 396. The compound (0.8 mg) was isolated in 2.9% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 98.0%; measured mass: 557.21; retention time: 1.56 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 557.17; retention time: 2.24 min.

Example 410 2,7-dicarbonitrile

Example 410 was prepared following the general procedure used for the synthesis of Example 396. The compound (1.9 mg) was isolated in 7.6% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 98.7%; measured mass: 510.22; retention time: 1.83 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 98.4%; measured mass: 510.19; retention time: 1.36 min.

Example 411 8-{4-[(4-Fluorophenyl)(phenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro-1,5naphthyridine-2,7-dicarbonitrile

The compound of Example 411 was prepared according to the general procedure used for the synthesis of the compound of Example 52. The compound (11.6 mg) was isolated in 48.5% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 97.9%; measured mass: 479.2; retention time: 1.48 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5)

- 233 044500 acetonitrile: water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 479.21; retention time: 2.22 min.

Example 412 ,5-naphthyridine-3-carbonitrile

Example 412 was prepared from racemic 1-((4-fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl)piperazine following the general procedure used for the synthesis of Example 395. The compound (8.4 mg) was isolated in 19 ,8%. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for 1st sample injection: Column: Waters XBridge

C18, 2.1 mmx50 mm, particle size 1.7 microns; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 532.2; retention time: 1.54 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 532.18; retention time: 2.3 min.

Example 413 ,5-naphthyridine-3-carbonitrile

With; lF ( ₄₁₃ )

Example 413 was prepared from homochiral 1-((4-fluoro-2-methoxyphenyl)(4fluorophenyl)methyl)piperazine (intermediate 110) following the general procedure used for the synthesis of Example 395. Compound (13.8 mg) was isolated with a yield of 60.4%. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 532.18; retention time: 2.32 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 532.19; retention time: 1.53 min.

Example 414 7-dicarbonitrile

- 234 044500

The compound of Example 414 was prepared according to the general procedure used for the synthesis of the compound of Example 52. The compound (10 mg) was isolated in 63.1% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 96.9%; measured mass: 495.12; retention time: 2.33 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 97.4%; measured mass: 495.14; retention time: 1.62 min.

Example 415 5-Methyl-8-{4-[(naphthalene-1-yl)methyl]piperazin-1-yl}-7-nitro-6-oxo-5,6-dihydro1,5-naphthyridine-2-carbonitrile

The compound of Example 415 was prepared according to the general procedure used for the synthesis of the compound of Example 18. The compound (10.1 mg) was isolated in 39% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 99.0%; measured mass: 455.13; retention time: 1.34 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 99.0%; measured mass: 455.13; retention time: 2.31 min.

Example 416 5-naphthyridine-2-carbonitrile

CH ₃

'3 ^sn s (416)

Example 416 was prepared according to the general procedure used for the synthesis

- 235 044500 compound of example 18. The compound (21.7 mg) was isolated in 52.6% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 543.2; retention time: 2.48 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 543.22; retention time: 1.64 min.

Example 417 6-Chloro-4-{4-[(4-chlorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile

To a solution of 6-chloro-1-methyl-2-oxo-4-(piperazin-1 -yl)-1,2-dihydro-1,5-naphthyridine-3-carbonitrile 2 HCl (15 mg, 0.040 mmol) in DMF (1.5 ml), 4-chlorobenzaldehyde (8.40 mg, 0.060 mmol) was added. This reaction mixture was stirred for 1 hour at room temperature. Sodium cyanoborohydride (7.51 mg, 0.119 mmol) was added and the reaction mixture was stirred overnight at room temperature. The reaction mixture was diluted with methanol, filtered and a light yellow solid was obtained as the final reaction product (2.6 mg, 5.77 µmol, yield: 14.5%). Analytical LC-MS conditions: column: Phenomenex LUNA C18, 2.0x50 mm, particle size 3 µm; mobile phase A: (10:90) methanol:water with 0.1% TFA; mobile phase B: (90:10) methanol:water with 0.1% TFA; gradient: 0100% B for 4 min, then maintain 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: retention time = 2.6 min; observed adducts: [M+H];

measured masses: 427.9. Analytical HPLC conditions: column: XTERRA 3.0x50 mm s7; mobile phase A: (10:90) acetonitrile:water with 0.1% TFA; mobile phase B: (90:10) acetonitrile:water with 0.1% TFA; gradient: 0-100% B for 4 min, then maintaining 100% B for 2 min; flow rate: 5 ml/min; registration: UV, 220 nm. HPLC results: purity: 100%.

Example 418

CH ₃ dXX NC NV CN 0„ 0¾ (418)

The compound of Example 418 was prepared according to the general procedure used for the synthesis of the compound of Example 52. The compound (1.2 mg) was isolated in 12% yield. Analytical LC-MS conditions: column: Phenomenex LUNA C18, 2x50 mm, particle size 3 µm; mobile phase A: 5% acetonitrile: 95% water: 10 mM ammonium acetate; mobile phase B: 95% acetonitrile: 5% water: 10 mM ammonium acetate; temperature: 40°C; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: retention time = 3.2 min; observed adducts: [M+H]; measured masses: 477.1.

Example 419 6-Chloro-4-{4-[(2-chloro-6-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile

- 236 044500

Example 419 was prepared following the general procedure used for the synthesis of Example 417. The compound (6.8 mg) was isolated in 28% yield. Analytical LCMS conditions: column: Phenomenex LUNA C18, 2x50 mm, particle size 3 µm; mobile phase A: 5% acetonitrile: 95% water: 10 mM ammonium acetate; mobile phase B: 95% acetonitrile: 5% water: 10 mM ammonium acetate; temperature: 40°C; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: retention time = 3.7 min; observed adducts: [M+H]; measured masses: 442.0.

Example 420 6-Chloro-4-{4-[(2-chloro-6-fluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile

Example 420 was prepared following the general procedure used for the synthesis of Example 417. The compound (10.1 mg) was isolated in 42.7% yield. Analytical LCMS conditions: column: Phenomenex LUNA C18, 2.0x50 mm, particle size 3 µm; mobile phase A: (10:90) methanol:water with 0.1% TFA; mobile phase B: (90:10) methanol:water with 0.1% TFA; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: retention time = 2.4 min; observed adducts: [M+H]; measured masses: 445.9.

Example 421 6-Chloro-4-{4-[1-(4-fluorophenyl)cyclopropyl]piperazin-1-yl}-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridine-3-carbonitrile

Example 421 was prepared from 4,6-dichloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3carbonitrile according to the general procedure used for the synthesis of Example 193. Compound (14.1 mg) was isolated in 54.5% yield. Analytical LC-MS conditions: column: Phenomenex LUNA C18, 2 mmx50, particle size 3 μm; mobile phase A: 5% acetonitrile: 95% water: 10 mM ammonium acetate; mobile phase B: 95% acetonitrile: 5% water: 10 mM ammonium acetate; temperature: 40°C; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: retention time = 3.5 min; observed adducts: [M+H]; measured masses: 438.0.

Example 422 6-Chloro-4-{4-[(2,6-difluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro1,5-naphthyridine-3-carbonitrile

- 237 044500

Example 422 was prepared following the general procedure used for the synthesis of Example 417. The compound (7.3 mg) was isolated in 30% yield. Analytical LCMS conditions: column: Phenomenex LUNA C18, 2.0x50 mm, particle size 3 µm; mobile phase A: (10:90) methanol:water with 0.1% TFA; mobile phase B: (90:10) methanol:water with 0.1% TFA; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: retention time = 2.2 min; observed adducts: [M+H]; measured masses: 429.9.

Example 423 6-Chloro-4-{4-[(2-fluoro-4-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile

Example 423 was prepared following the general procedure used for the synthesis of Example 417. The compound (9.4 mg) was isolated in 40.3% yield. Analytical LCMS conditions: column: Phenomenex LUNA C18, 2 mmx50, 3 µm particles; mobile phase A: 5% acetonitrile: 95% water: 10 mM ammonium acetate; mobile phase B: 95% acetonitrile: 5% water: 10 mM ammonium acetate; temperature: 40°C; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: retention time = 3.2 min; observed adducts: [M+H]; measured masses: 426.1.

Example 424 -carbonitrile

The compound of Example 424 was prepared according to the general procedure used for the synthesis of the compound of Example 417. The compound (10.2 mg) was isolated in 41.5% yield. Analytical LCMS conditions: column: Phenomenex LUNA C18, 2x50 mm, particle size 3 µm; mobile phase A: 5% acetonitrile: 95% water: 10 mM ammonium acetate; mobile phase B: 95% acetonitrile: 5% water: 10 mM ammonium acetate; temperature: 40°C; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: retention time = 2.9 min; observed adducts: [M+H]; measured masses: 449.1.

Example 425 -naphthyridine-3-carbonitrile

- 238 044500

Example 425 was prepared from 4,6-dichloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3carbonitrile according to the general procedure used for the synthesis of Example 193. Compound (11.8 mg) was isolated in 52.0% yield. Analytical LC-MS conditions: Column: Phenomenex

LUNA C18, 2.0x50 mm, particle size 3 µm; mobile phase A: (10:90) methanol:water with 0.1% TFA; mobile phase B: (90:10) methanol:water with 0.1% TFA; gradient: 0-100% B for 4 min, then maintaining 100% B for 1 min; flow rate: 0.8 ml/min; registration: UV, 220 nm. LC-MS results: retention time = 3.0 min; observed adducts: [M+H]; measured masses: 439.9.

Example 426

4-Chloro-1-methyl-1,5-naphthyridin-2(1H)-one (12.3 mg, 0.063 mmol) was dissolved in DMF (632 μl) to obtain a solution. 1-(4,4'-difluorobenzhydryl)piperazine (22.9 mg, 0.079 mmol) and potassium carbonate (18.7 mg, 0.135 mmol) were then added. This reaction mixture was placed under a nitrogen atmosphere and heated for 18 hours at 80°C. HPLC analysis showed approximately 50% conversion of the starting materials into the reaction product. Potassium carbonate (8.2 mg, 0.059 mmol) was added to the reaction mixture. The reaction mixture was sealed under a nitrogen atmosphere and heated for 19 hours at 85°C. The resulting crude material was purified by preparative LC-MS using the following conditions: column: XBridge C18, 19x200 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: 5090% B for 15 min, then maintain 100% B for 5 min; flow rate: 20 ml/min. Fractions containing the desired product were combined and dried using a centrifugal evaporator. The product yield was 21.1 mg and its purity, determined by LC-MS analysis, was 100%. To determine the final purity of the product, the sample was analyzed twice using analytical LC-MS. Analysis conditions for the 1st sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Analysis conditions for the 2nd sample injection: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. LC-MS: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Retention time = 1.42 min; observed adducts: [M+H]; measured masses: 447.1. LC-MS: column: Waters Acquity UPLC VEN C18, 2.1x50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: 0-100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1.0 ml/min; registration: UV, 220 nm. Retention time = 2.25 min; observed adducts: [M+H]; measured masses: 447.1. Proton NMR signal intensities near the frequency used to suppress the water signal are distorted and uncorrected. ¹ H NMR (DMSO-d ₆ ) δ 8.42 (d, J=2.9 Hz, 1H), 7.91 (d, J=8.8 Hz, 1H), 7.57 (dd, J=8.8, 4.4 Hz, 1H), 7.48 ( dd, J=8.6, 5.7 Hz, 4H), 7.14 (t, J=8.8 Hz, 4H), 5.98 (s, 1H), 4.47 (s,

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1H), 3.51 (s, 2H).

Example 427 4-{4-[bis(4-Fluorophenyl)methyl]piperazin-1-yl}-1,6-dimethyl-1,2-dihydro-1,5-naphthyridin-2-one

2nd Generation RuPhos Precatalyst (CAS No. 1375325-68-0; 27.9 mg, 0.036 mmol), 4-chloro-1,6dimethyl-1,5-naphthyridin-2(1H)-one (150 mg, 0.719 mmol ), 1-(4,4'-difluorobenzhydryl)piperazine (249 mg, 0.863 mmol) and cesium carbonate (703 mg, 2.157 mmol) were mixed in 7.1 ml of solvent (DMA/tert-BuOH; 1:4) in a test tube volume 20 ml. This mixture was sealed under a nitrogen atmosphere and heated for 72 hours at 90°C. According to LC-MS analysis, the reaction was complete. Volatile components were removed under vacuum using a system consisting of a rotary evaporator and a vacuum pump. The residue was suspended in DCM and CHCl ₃ and filtered through a pad of diatomaceous earth (Celite). The mixture was then adsorbed onto 1.3 g of silica gel and chromatographed using a suspension of silica gel (13.6 g) in 15% ethyl acetate in dichloromethane and eluting with 15% ethyl acetate in dichloromethane. After drying under vacuum, the title compound (217 mg) was obtained in 62.3% yield. 1H NMR (chloroform-d) δ 7.56 (d, J=8.7 Hz, 1H), 7.38-7.43 (m, 4H), 7.29 (d, J=8.7 Hz, 1H), 6.97-7.03 (m, 4H), 6.16 (s, 1H), 4.31 (s, 1H), 3.62 (s, 3H), 3.57 (br. s., 4H), 2.63 (t, J=4.8 Hz, 4H), 2.57 (s, 3H).

Example 428

6-bromo-4-{4-[(S)-(4-chlorophenyl)(phenyl)methyl]niperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridin-3 -carbonitrile

The compound of Example 428 was prepared from (S)-1-((4chlorophenyl)(phenyl)methyl)piperazine according to the general procedure used for the synthesis of the compound of Example 393. The compound (23 mg) was isolated in 41.9% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 548.07; retention time: 2.59 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 548.06; retention time: 1.72 min.

Example 429 3-carbonitrile

- 240 044500

Example 429 was prepared from racemic 1-((4-chlorophenyl)(phenyl)methyl)piperazine following the general procedure used for the synthesis of Example 393. The compound (29 mg) was isolated in 52.8% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 548.09; retention time: 1.73 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 548.06; retention time: 2.56 min.

Example 430 3-carbonitrile

Example 430 was prepared from racemic 1-((4-fluorophenyl)(phenyl)methyl)piperazine following the general procedure used for the synthesis of Example 393. The compound (30.8 mg) was isolated in 57.8% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample input: purity: 100.0%; measured mass: 532.08; retention time: 2.41 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 100.0%; measured mass: 532.09; retention time: 1.58 min.

Example 431 3-carbonitrile

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Example 431 was prepared following the general procedure used for the synthesis of Example 393. The compound (12.1 mg) was isolated in 22.9% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 94.8%; measured mass: 528.15; retention time: 1.6 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 94.9%; measured mass: 528.09; retention time: 2.54 min.

Example 432 8-{4-[bis(4-Fluorophenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile

Example 432 was isolated as the main product in a trial synthesis (4-(4-(bis(4 fluorophenyl)methyl)piperazin-1-yl)-6-cyano-1-methyl-2-oxo-1,2-dihydro- 1,5-naphthyridin-3-yl)boronic acid. To a solution of 8-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-7-bromo-5-methyl-6-oxo-5,6-dihydro1,5-naphthyridine-2-carbonitrile (0.065 g, 0.118 mmol) in THF (1.181 ml) having a temperature of -78°C, a 2.5 M solution of n-butyllithium (0.052 ml, 0.130 mmol) was added dropwise. This reaction mixture was stirred for 1 hour at -78°C and then trimethyl borate (0.020 ml, 0.177 mmol) was added. The reaction mixture was stirred for 30 minutes at -78°C and then for 16 hours at room temperature. LC-MS analysis indicated the formation of a hydro-dehalogenated product. This crude substance was purified by preparative LC-MS using the following conditions: column: XBridge C18, 200 mmx19 mm, particle size 5 μm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; gradient: maintaining 37% B for 0 min, 37-77% B for 22 min, then maintaining 100% B for 5 min; flow rate: 20 ml/min; Column temperature: 25°C. Fraction collection was triggered by MS and UV signals. Fractions containing the desired product were combined and dried using a centrifugal evaporator. The compound (22.6 mg) was isolated in 40.6% yield. Analytical LC-MS was used to determine the final purity of the product. Analysis conditions for the 1st sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: (95:5) acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 1st sample injection: purity: 98.6%; measured mass: 472.18; retention time: 1.46 min. Analysis conditions for the 2nd sample injection: column: Waters XBridge C18, 2.1 mmx50 mm, particle size 1.7 µm; mobile phase A: (5:95) acetonitrile:water with 10 mM ammonium acetate; mobile phase B: (95:5) acetonitrile:water with 10 mM ammonium acetate; temperature: 50°C; gradient: from 0% B to 100% B for 3 min, then maintaining 100% B for 0.75 min; flow rate: 1 ml/min; registration: MS and UV (220 nm). Analysis results for the 2nd sample injection: purity: 97.3%; measured mass: 472.19; retention time: 2.23 min.

Analysis of biological activity.

The pharmacological properties of the compounds of the present invention can be confirmed by studies of their biological activity. The following are examples of biological activity assays in which the compounds of the invention are used.

1. In vitro DGK inhibition assay.

Reactions involving DGKa and DGKZ were performed using either extruded liposomes (LIPGLO assay for DGKa and DGKZ enzymes) or a detergent/lipid micellar substrate (assay for DGKa and DGKZ enzymes). These reactions were performed in assay buffer: 50 mM MOPS pH 7.5, 100 mM

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NaCl, 10 mM MgCl2, 1 μM CaCl2 and 1 mM DTT. Reaction mixtures using the detergent/lipid micellar substrate also contained 50 mM octyl B-D-glucopyranoside. In reactions with detergent/lipid micelles, the concentrations of the lipid components of the substrate were: 11 mM PS (phosphatidylserine) and 1 mM DAG (diacylglycerol). In reactions with extruded liposomes, the concentrations of substrate lipid components were: 2 mM PS, 0.25 mM DAG, and 2.75 mM PC (phosphatidylcholine). Reactions were performed in the presence of 150 μM ATP. The concentrations of DGKa and DGKZ enzymes were 5 nM.

The study of the inhibitory activity of the compounds was carried out as follows. Compounds were dissolved in DMSO and 50 nL drops of each test compound (10 mM initial concentration and 11 serial 3-fold dilutions for each compound) were transferred to the wells of a white 1536-well plate (Corning 3725). Prepare 5 mL of 2x working concentration enzyme/substrate solution by mixing 2.5 mL of 4x enzyme solution (20 nM DGKa or DGKZ solution prepared as described below in assay buffer ) and 2.5 ml of 4x liposome solution or 4x detergent/lipid micelles solution (formulations described below) and this solution was incubated for 10 min at room temperature. Then 1 μl of 2x enzyme/substrate solution was added to the wells containing the test compound and the reaction was started by adding 1 μl of 300 μM ATP solution. The reaction was continued for 1 h, then 2 μl of Glo reagent (Promega V9101) was added and incubated for 40 min. Then, 4 μl of kinase detection reagent was added and incubated for 30 min. Luminescence was recorded using an EnVision plate reader. Percentage inhibition was calculated from ATP conversion in control reactions in the absence of enzyme (100% inhibition) and in reactions using vehicle alone (0% inhibition). To determine the IC ₅₀ (the concentration required for 50% inhibition), 11 concentrations of the compound were analyzed.

4x-detergent/lipid micelles preparation.

Detergent/lipid micelles were prepared by mixing 15 g phosphatidylserine (Avanti 840035P) and 1 g diacylglycerol (800811O) and dissolving this mixture in 150 ml chloroform in a 2 L round bottom flask. Chloroform was removed under high vacuum by rotary evaporation. The resulting colorless sticky oil was resuspended in 400 ml of 50 mM MOPS pH 7.5, 100 mM NaCl, 20 mM NaF, 10 mM MgCl ₂ , 1 μM CaCl ₂ , 1 mM DTT and 200 mM octyl glucoside by vigorous stirring. The resulting lipid/detergent solution was divided into 5 ml aliquots and stored at -80°C.

4x-Liposome preparation.

Lipid composition: 5 mol % DAG (Avanti 800811O), 40 mol % PS (Avanti 840035P) and 55 mol % PC (Avanti 850457); the total lipid concentration in the 4x liposome solution was 15.2 mg/ml. PC, DAG and PS were dissolved in chloroform, mixed and dried under vacuum to obtain a thin film. Lipids were hydrated to 20 mM in 50 mM MOPS pH 7.5, 100 mM NaCl, 5 mM MgCl ₂ and frozen and thawed five times sequentially. The resulting lipid suspension was extruded eleven times through a polycarbonate filter with a pore size of 100 nm. The size of the liposomes (radius 50–60 nm) was confirmed using dynamic light scattering. The liposome preparation was stored at 4°C for no more than four weeks.

Baculovirus expression of human DGKa and DGKZ.

Baculovirus samples (DGK-human alpha)-TVMV-His-pFBgate and (DGK-human zeta transcript variant 2)-TVMV-His-pFBgate were prepared using the Bac-to-Bac baculovirus expression system (Invitrogen) according to the manufacturer's protocol . The DNA used to express DGK-alpha and DGK-zeta had the sequences SEQ ID NO: 1 and 3, respectively. Baculovirus amplification was carried out by infecting Sf9 cells at a virus/cell ratio of 1:1500 and growing after transfection for 65 h at 27°C.

Large-scale expression of each protein was performed in a Cellbage 50L WAVE-Bioreactor System 20/50 (GE Healthcare Bioscience). Sf9 cells (12 L, 2x106 cells/ml) (Expression System, Davis, CA), grown in ESF921 insect cell culture medium (expression system), were infected with the viral strain at a virus/cell ratio of 1:200 and grown after infection for 66-68 hours at 27°C. The infected cell culture was collected by centrifugation at 2000 rpm for 20 min at 4°C in a SORVALLZ RC12BP centrifuge. Cell pellets were stored at -70°C until purification.

Purification of human DGKa and DGKZ.

Each of the expressed full-length human DGKa and DGKZ enzymes containing the TVMV protease-cleavable C-terminal marker sequence Hexa-His (SEQ ID NO: 2 and 4, respectively) and obtained as described above, was purified from the cell mass of baculovirus-infected Sf9- insect cells. Cells were lysed using a nitrogen cavitation technique using a pressurized nitrogen balloon (Parr Instruments), and the resulting lysates were clarified by centrifugation. Clarified lysates were purified to ~90% homogeneity using three sequences.

- 243 044500 operational stages of column chromatography on the AKTA Purifier Plus system. These three column chromatography steps included affinity retention on a nickel resin (namely HisTrap FF crude, GE Healthcare) and then size exclusion chromatography (namely HiLoad 26/600 Superdex 200 prep grade, GE Healthcare for DGK-alpha and HiPrep 26/600 Sephacryl S 300_HR, GE Healthcare for DGK-zeta). The third step was ion exchange chromatography and differed between the two isoforms. DGKa was further purified using anion exchange chromatography on Q-Sepharose (GE Healthcare). DGKZ was further purified using SP-Sepharose cation exchange chromatography (GE Healthcare). The resulting proteins had a concentration of >2 mg/ml. The compositions of the buffer solutions for both proteins were identical: 50 mM Hepes, pH 7.2, 500 mM NaCl, 10% v/v. glycerol, 1 mM tCeP and 0.5 mM EDTA.

2. Analysis of IL-2 (interleukin-2) production by Raji cells and CD4 T cells.

The 1536-well IL-2 assay was performed in a 4-μl volume using preactivated CD4 T cells and Raji cells. Prior to the assay, CD4 T cells were preactivated by treatment with a-CD3, a-CD28, and PHA at concentrations of 1.5, 1, and 10 μg/ml, respectively. Raji cells were treated with staphylococcal enterotoxin B (SEB) at a concentration of 10,000 ng/ml. First, serially diluted compounds were transferred into a 1536-well assay plate (Corning, #3727) and then 2 μl of preactivated CD4 T cells (final density 6000 cells/well) and 2 μl of SEB-treated Raji cells (2000 cells/well) were added. hole). After incubation for 24 hours in an incubator at 37°C/5% CO2, 4 μl of IL-2 detection reagent (Cisbio, #64IL2PEC) was added to the assay plate. The assay plate was read using an Envision plate reader. To assess the cytotoxicity of the compounds, either Raji cells or CD4 T cells were incubated with serially diluted compounds. After incubation for 24 h, 4 μl of CellTiter Glo (Promega, #G7572) was added and the plates were read using an Envision plate reader. The 50% effective concentration (IC ₅₀ ) was calculated using the four-parameter logistic formula y = A+((BA)/(1+((C/x) ^A D))), where A and B mean the minimum and maximum % activation or inhibition, respectively, C is the IC ₅₀ , D is the Hill slope, and x is the concentration of the compound.

3. CellTiter-Glo CD8 T cell proliferation assay.

Frozen naive human CD8 T cells were thawed in RPMI medium with 10% FBS (fetal bovine serum), incubated for 2 hours at 37°C, and cell concentrations were determined. A 384 well tissue culture plate was coated overnight at 4°C with anti-human CD3 antibody (20 μl; 0.1 μg/ml in unsupplemented RPMI), then the antibody solution was removed from the plate and CD8 T cells were added to each well ( 20,000/40 µl) together with soluble anti-human CD28 antibody (0.5 µg/ml). Immediately after the cells were added to the cell plate, compounds were added. After incubation for 72 hours in a 37°C incubator, 10 μl of CellTiterglo reagent (Promega, Catalog No. G7570) was added to each well. The plate was shaken vigorously for 5 min, incubated at room temperature for an additional 15 min, and read using Envision to assess CD8 T cell proliferation. In this assay, the signal from CD8 T cells stimulated with anti-CD3 (0.1 μg/ml) and anti-CD28 (0.5 μg/ml) was considered background. Example 40 (at a concentration of 3 μM) was used to normalize the obtained data, taking its effect as 100% and considering its EC ₅₀ to correspond to an absolute 50% effect.

4. Analysis using the DGK AP1 reporter.

The AP1-luciferase reporter in the Jurkat cell line was prepared using the Cignal Lenti API Reporter (luc) Kit from SABiosciences (CLS-011L).

Compounds were transferred from the Echo LDV plate to individual wells of a 384-well plate (white, hard bottom, opaque PE CulturPlate 6007768) using an Echo550 apparatus. The sample volume was 30 nL per well, and one receiver plate was used for each source plate. Cell suspension was prepared by transferring 40 ml of cells (2x20 ml) into empty 50 ml conical tubes. Cells were concentrated by centrifugation (1200 rpm) for 5 min at ambient temperature. The supernatant was removed and the cells were suspended in RPMI medium (Gibco 11875) with 10% FBS to a density of 1.35 x 10 ⁶ cells/ml. Cells were added manually using a multichannel pipette into a 384-well TS plate containing compounds, adding 30 μl of cell suspension (4.0 x 10 ⁴ cells) to each well. Cell plates were incubated for 20 min at 37°C in an atmosphere of 5% CO ₂ .

While incubation was in progress, anti-CD3 antibody (aCD3) solutions were prepared by mixing 3 μl of aCD3 (1.3 mg/ml) with 10 ml of medium (final concentration = 0.4 μg/ml). Then, 1.5 μl of aCD3 (1.3 mg/ml) was mixed with 0.5 ml of medium (final concentration = 4 μg/ml). After 20 minutes, 10 μl of medium was added to all wells A - M of column 1 and 10 μl of aCD3 (4 μg/ml) was added to the wells of column 1, rows N - P, for control. Then, 10 μL of aCD3 (0.4 μg/mL) per well was added using a multichannel pipette. αCD3-stimulated cells, treated or untreated with compounds, were incubated at 37°C, in an atmosphere of 5% CO2 for 6 hours.

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During this incubation period, Steady-Glo reagent (Promega E2520) was slowly thawed at ambient temperature. 20 µl of Steady-Glo reagent per well was then added using a Multidrop Combi dispenser. Gas bubbles were removed by centrifugation (2000 rpm) for 10 s at ambient temperature. Cells were incubated for 5 min at room temperature. Measurements characterizing the samples were obtained using the Envision Plate Reader equipment and luminescence protocol and are expressed in relative light units (RLU). The data obtained were analyzed using the compound of example 40 for normalization and taking its inhibition to be 100%.

5. Analysis of mouse cytotoxic T lymphocytes.

The antigen-specific cytolytic T cell (CTL) assay was developed to functionally evaluate the ability of inhibitors of the enzymes DGKa and DGKZ to enhance effector T cell-mediated tumor cell killing activity. CD8+ T cells isolated from OT-1 transgenic mice recognize antigen-presenting cells MC38, which present the ovalbumin-derived peptide SIINFEKL. Recognition of cognate antigen initiates the cytolytic activity of antigen-specific CD8+ T cells from OT-1 mice.

Functional CTL cells were prepared as follows. Splenocytes were isolated from 8-12 week old OT-1 mice and expanded in the presence of SIINFEKL peptide (1 μg/ml) and mIL2 (10 U/ml). After three days, fresh medium containing mIL (2 units/ml) was added. On the fifth day, the resulting CD8+ T cells were isolated and prepared for use. Activated CTL cells can be stored frozen for 6 months. In addition, one million MC38 tumor cells were vibrated in the presence of SIINFEKL-OVA peptide (1 μg/ml) for 3 h at 37°C. Cells were washed (3X) with fresh medium to remove excess peptide. Finally, CTL cells, pretreated with DGK inhibitors for 1 h in a 96-well U-bottom plate, were mixed at a 1:10 ratio with antigen-loaded MC38 tumor cells. The cells were then centrifuged (700 rpm) for 5 min and placed overnight in an incubator at 37°C. After 24 h, the supernatant was collected, and IFN-γ cytokine levels were determined using an AlphaLisa immunoassay kit obtained from Perkin Elmer.

6. RNA analysis of proliferation.

Phytohemagglutinin (PHA)-stimulated blast cells from frozen samples were incubated in RPMI medium (Gibco, ThermoFisher Scientific, Waltham, MA) supplemented with 10% fetal calf serum (Sigma Aldrich, St. Louis, MO) for one hour and then added to individual wells of a 384-well plate (10,000 cells per well). Compounds were then transferred to individual wells of a 384-well plate and the treated cells were incubated for 72 hours at 37°C in an atmosphere of 5% CO ₂ in culture medium containing human IL2 (20 ng/ml) and then cell growth was measured using MTS reagent (3-(4,5-dimethyl-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)2H-tetrazolium) following the manufacturer's instructions (Promega, Madison, WI). Percentage inhibition was calculated by comparison with IL2-stimulated (0% inhibition) and unstimulated control (100% inhibition). The values of inhibitory concentrations (IC ₅₀ ) were calculated on the basis of 50% inhibition, taking into account the multiplicity of induction when treating stimulated and unstimulated cells with IL2.

7. Analysis of IFN-γ production by human CD8 T cells.

Frozen naïve human CD8 T cells were thawed in AIM-V medium, incubated for 2 hours at 37°C, and cell concentrations were determined. A 384-well tissue culture plate was coated overnight at 4°C with anti-human CD3 antibody (20 μl; 0.05 μg/ml in PBS), then the antibody solution was removed from the plate and 40,000 CD8 was added to each well. T cells in a 40 µl volume containing soluble anti-human CD28 antibody (0.1 µg/ml). Immediately after adding cells, compounds were added to the cell plate using an Echo liquid handler. After incubation for 20 h in a 37°C incubator, 3 μl of supernatant from each well was transferred to a new 384-well white assay plate and cytokine levels were measured.

To quantify interferon γ (IFN-γ), the AlphLISA Kit (Catalog No. AL217) was used as described in the manufacturer's manual (Perkin Elmer). The signal intensity from each well was converted into IFN-γ concentration (pg/ml). The EC ₅₀ values of the compounds were determined using co-stimulation with anti-CD3 (0.05 μg/ml) and anti-CD28 (0.1 μg/ml) as baseline and co-stimulation with the example compound as 100% activation. 40 (3 µM) together with anti-CD3 antibody and anti-CD28 antibody.

8. Analysis of pERK production by CD8 human T cells.

Frozen naïve human CD8 T cells were thawed in AIM-V medium, incubated for 2 hours at 37°C, and cell concentrations were determined. Positive CD8 T cells were added to a 384-well tissue culture plate at 20,000 cells per well in AIM-V medium. One compound was added to each well, then bead-bound anti-human CD3 and anti-CD28 monoclonal antibodies (mAbs) were added to a final concentration of 0.3 μg/ml.

- 245 044500 cells were incubated for 10 min at 37°C. The reaction was stopped by adding lysis buffer from the AlphaLISA Surefire Kit. (Perkin Elmer, Catalog No. ALSU-PERK-A). Lysates (5 μl per well) were transferred to a new 384-well white plate to measure pERK activation.

The EC ₅₀ values of the compounds were determined using co-stimulation with anti-CD3 and anti-CD28 as baseline and co-stimulation with Example 40 (3 μM) plus anti-CD3 and anti-CD28 as 100% activation.

9. Analysis of IFN-γ levels in human whole blood.

Human venous whole blood obtained from healthy donors (22.5 μl per well) was pretreated with compounds for one hour at 37°C in a humidified incubator under a 95% air/5% CO2 atmosphere. Blood was stimulated by adding 2.5 μl of human anti-CD3 and anti-CD28 monoclonal antibodies (mAbs) to a final concentration of 1 μg/ml of each antibody and incubating for 24 h at 37°C. The level of IFN-γ in the supernatants was measured using the AlphLISA Kit (Catalog No. AL217).

The EC ₅₀ values of the compounds were determined using co-stimulation with anti-CD3 and anti-CD28 as baseline and co-stimulation with Example 40 (3 μM) plus anti-CD3 and anti-CD28 as 100% activation.

Table 1. IR ₅ o activity values in in vitro DGK inhibition assay

EXAMPLE No, DGKa LIPGLO IC50 (µM) ϋΟΚζ LIPGLO IR50 (µM) DGKa IC50 (µM) ΌΟΚζ IR ₅₀ (µM) 2 - - 0.049 >240 3 3.8 2.3 2.1 >240 6 - - 6.5 >240 7 - - θ,5 120 8 0.41 5.4 0.16 20 9 - - 3.8 >240 10 0.31 3.9 0.12 23 AND 1.8 1.8 0.45 4 12 - - 1 >81 13 5.4 - 9.5 >240 14 4.7 3.2 1.2 >240

- 246 044500

15 - - 0.55 58 16 4.3 27 6.4 >240 17 3.5 9.4 0.93 180 18 - - 0.96 2.5 19 - - 13 19 20 - - 3.3 4.3 21 - - 22 94 22 - - 0.07 >81 23 1.9 13 0.61 >240 24 0.15 90 0.034 >240 25 - - 0.041 >81 26 6.7 18 4.3 85 27 ο,ιι >240 ο,ιι >240 28 - - 1.3 >81 29 - - 0.57 >240 thirty - - 1.8 140 31 - - 1.2 >240 32 - - 0.19 9.4 33 1.7 1.7 0.75 7.6 34 0.29 3.4 0.18 23 35 0.99 81 0.14 >240 36 6.8 18 1.4 68 37 - - 1.8 38 39 - - 50 >240 40 3.4 0.21 1.9 1.3 43 - - 0.037 6.3 44 0.37 1.3 0.23 8.9 45 0.39 6.8 ο,ι 36 46 - - 8.5 2.7 47 - - 1.8 >81 48 3.9 4.5 >240 >240 49 - - 0.18 >240 50 2.1 5.7 0.99 39 51 0.33 4.7 0.18 56 52 - - 7.8 150 53 - - 1.5 16 54 3.1 1.3 0.82 7.3 55 0.19 1.8 - - 57 - - 1.6 110 58 - - 0.25 19 60 - - 32 >240 61 - - 4.4 2.2 64 - - 2.7 14

- 247 044500

65 0.51 3.7 0.29 20 66 0.23 1.7 0.084 13 67 4.8 12 0.13 13 68 - - 4 23 69 - - 1.3 13 70 - - 0.13 3.3 73 - - 0.9 2.1 75 - - 2.6 130 76 - - 5.5 2.5 78 - - 12 >240 79 - - 81 >81 85 - - 7.6 0.77 89 - - 41 >240 90 >240 15 82 190 95 - - >240 >240 96 - - 87 53 98 - - 6.7 4.4 99 0.72 0.04 0.41 0.67 101 - - 15 27 103 6.1 0.99 - - 104 0.74 0.78 - - 105 - - 0.58 2.5 106 - - 0.96 2 114 - - 120 100 115 - - 220 >240 116 - - >240 >240 117 - - 46 21 118 1.3 0.061 4.2 0.23 119 - - 0.97 0.19 120 - - 18 0,>81 121 6.4 0.055 1.9 0.18 122 - - 1 0.72 123 - - 2.3 3 124 0.75 0.16 0.75 0.9 125 AND 2.3 4.3 7.9 126 - - 6.1 0.33 127 2.9 0.03 4.2 0.25 128 18 0.048 21 1.5 129 - - 4.2 0.18 130 - 0.31 thirty 1.9 131 - - AND 0.37 132 - - 3.3 0.15 133 - - 3.3 0.14

- 248 044500

134 - - 31 4.2 135 - - 0.18 1.3 136 0.79 0.21 0.21 0.87 137 1.5 0.85 0.28 4.8 138 - - 32 0.73 139 - - 0.41 0.91 140 2.7 0.13 0.64 0.6 141 0.58 1.4 0.27 8.9 143 3.4 1.1 1.3 4.2 144 - - 0.53 7.9 145 - - thirty 2.3 146 - - >240 36 147 - - 76 3.6 148 - - 15 28 149 - - 47 86 150 - - 73 88 151 7.3 3.1 38 27 152 - - 150 3.9 153 - - 180 76 154 81 72 45 >240 155 71 63 24 230 156 - - 170 140 157 - - 53 26 158 - - >240 >240 159 - - 120 >240 160 - - 180 160 161 - - >240 >81 162 - - 160 >240 163 110 110 >240 >240 164 81 59 >240 >240 165 2.8 4.1 3.4 92 166 210 230 - - 176 >240 27 - - 177 >240 9.7 - - 183 >240 21 52 >240 186 - - >240 >240 189 - - 130 >240 190 - - 58 >240 192 0.048 8.1 0.084 >240 193 0.089 6.5 0.22 >240 194 0.18 8.3 0.076 62 195 1.7 98 0.22 >240 196 2.7 43 4 >240

- 249 044500

198 - - 0.074 150 199 - - 0.48 >240 200 - - 24 >240 202 - - 31 12 211 - - 0.25 >240 212 - - 2.2 >240 213 - - 3.1 >240 214 - - 3.1 >81 215 - - 4.1 >81 216 1.2 4.8 1.4 54 217 1.3 1.4 2.3 4.5 218 1.5 5.1 0.69 >81 220 - - 1.7 89 221 3.4 18 0.74 >240 222 - - 0.083 - 223 9.9 0.62 1.9 3.2 224 0.14 20 0.54 >240 225 0.82 1.3 0.39 14 226 3.9 10 1.6 70 227 230 AND >240 >240 228 >240 9.7 - >240 229 >240 AND 14 14 230 - - 8.8 >240 231 - - 170 >240 232 - - 150 >240 233 3 6.6 8.4 >240 234 - - 16 - 239 9.9 51 AND >240 244 - - 32 >81 250 5.1 >240 - - 251 >240 9 - - 252 >240 200 - - 253 40 7.4 - - 254 1.7 16 - - 255 1.4 21 - - 256 2.7 10 - - 258 1.6 5.3 0.43 26 259 0.49 4 0.42 55 260 1.6 5.3 0.35 - 261 0.71 10 0.43 >240 262 - - 0.52 >240 263 0.22 5.2 0.084 14 264 - - 0.62 >240

- 250 044500

265 - - 0.034 36 266 0.022 And 0.027 >240 267 - - 0.31 >240 268 - - 0.68 120 269 0.21 2.8 0.052 26 270 - - 0.19 >240 271 - - 1.1 110 272 2.3 10 1.5 >240 273 0.082 7.8 0.018 69 274 1.6 5.4 0.67 thirty 275 0.77 10 o, s 58 276 - - 0.012 >240 277 1.7 13 0.75 45 278 - - 0.35 >81 279 0.81 5.8 0.38 56 280 - - 0.076 >240 281 2 9.9 0.39 49 282 - - 0.22 >240 283 0.21 3 0.12 27 284 - - 1 >240 285 - - 0.91 >240 286 - - 1.2 140 287 18 29 2.1 >240 288 0.033 2.3 0.037 >240 289 - - 0.25 >81 290 - - 2.2 >240 291 - - 0.33 >81 292 0.48 2.4 0.27 25 293 0.6 6 0.27 58 294 - - ο,ιι >240 295 0.16 6.9 0.13 55 296 - - 0.98 150 297 - - 0.89 >240 298 - - 0.83 27 299 - - 0.57 >240 300 24 98 2 >240 301 - - 0.068 >240 302 - - 0.61 >81 303 - - 0.9 >240 304 0.61 5.8 0.33 68 305 0.49 7.5 0.13 62 306 - - 0.2 >240 307 - - 1.8 >81

- 251 044500

308 0.95 13 0.19 - 310 - - 0.97 >240 312 0.22 16 0.12 >240 313 17 62 1.5 >81 314 - - 0.17 82 315 - - 1.1 >240 315 >240 1.6 >240 >240 316 0.25 >240 0.4 >240 317 - - 0.16 34 318 - - 0.85 140 319 - - 0.55 220 320 - - 0.89 >240 321 0.34 12 0.16 >240 322 - - 2.8 >240 323 1.2 27 0.14 AND 324 - - 0.19 >240 326 - - 1.9 >240 327 - - 0.66 140 328 - - 1 120 329 5.6 49 0.56 thirty 330 - - 3.7 >240 331 - - 0.49 >240 332 2.5 6.5 1.1 67 333 - - 5 230 334 1.7 27 - - 335 1.9 18 2.7 >240 337 - - 0.065 >81 338 5.1 66 1.1 >240 339 - - 0.57 >240 340 3.4 17 4.2 >240 341 - - 0.32 >240 342 - - 28 >240 343 - - 0.068 >81 344 - - 0.4 >240 345 - - 4.6 >240 346 - - 5.3 >240 347 2.6 - - - 348 - - 0.22 140 349 1.3 32 - - 350 - - 1.2 >81 351 - - 0.36 190 352 - - 0.7 >81 353 - - 0.065 >240

- 252 044500

354 - - ο,ιι >240 355 - - 7.1 >240 356 - - o, s >240 357 0.085 47 0.028 >240 358 >240 >240 - - 359 - - 19 >240 360 - - 48 >240 362 - - 0.069 >240 363 - - 0.15 >240 364 - - 1.1 >240 365 - - 3.5 190 366 - - 1.3 160 367 - - 0.22 >240 368 - - 0.4 >240 369 - - 0.16 >240 370 - - 0.17 >240 371 - - 0.23 170 372 0.078 8.8 0.033 60 373 - - 2.5 100 375 - - 1.6 >240 377 >240 130 - - 378 2.2 >240 - - 379 - - 0.38 >240 380 - - 0.83 >240 381 0.073 27 0.018 >240 382 0.76 >240 - - 383 0.22 >240 - - 385 - - 0.35 150 386 0.26 6.7 0.15 190 387 - - 0.0041 >240 387 0.27 1.6 0.46 >240 388 0.051 130 0.053 >240 389 1.4 9.4 0.85 130 390 - - 0.87 >240 391 1.8 2.1 0.26 36 392 >240 2.3 - - 393 9 >240 - - 394 3.9 1.2 - - 395 14 3 - - 396 21 5.8 - - 397 16 13 - - 398 0.097 - - - 398 0.21 >240 - -

- 253 044500

398 1.7 49 - - 399 6.4 0.16 - - 400 5.4 2.1 - - 401 81 7.6 - - 402 160 3.5 - - 403 26 5.7 - - 404 4.9 3.9 - 63 405 0.56 0.86 - - 406 13 3.1 - - 407 0.96 0.45 - - 408 9 1.1 - - 409 >240 0.95 - - 410 5.4 2.2 - - 411 4.9 1.4 - - 412 1.8 0.6 - - 413 z, z 0.97 - - 414 9 1.5 - - 415 0.65 0.66 - - 416 4.8 0.91 - - 417 - - 0.25 >240 419 49 15 81 >81 421 - - 69 >240 422 - - 1.4 >240 423 2.2 33 1.4 >81 424 1.4 27 0.77 >240 425 20 >240 1.8 >240 426 170 >240 - - 427 - - 150 >240 428 10 5.2 - - 429 1.8 9.4 - - 430 1.9 3 - - 431 9 9 - - 432 >240 5.4 - -

Table 2. IC5o activity values in the analysis of IL-2 production by Raji cells and CD4 T cells

EXAMPLE No. RAJI IC50 (µM) EXAMPLE No. RAJI IC50 (µM) 2 0.20 233 0.48 3 0.31 234 0.13

- 254 044500

6 0.82 239 3.6 7 0.075 244 0.69 8 0.091 258 0.0053 9 >100 259 0.016 10 0.0030 260 0.018 13 >100 261 0.021 14 0.026 262 0.028 16 0.021 263 0.035 18 0.023 264 0.036 19 >100 265 0.039 20 >100 266 0.041 21 0.41 267 0.048 23 0.041 268 0.051 24 0.012 269 0.057 26 0.14 270 0.059 27 0.057 271 0.061 29 2.3 272 0.064 thirty 1.1 273 0.064 31 1.8 275 0.073 33 0.018 276 0.079 34 0.035 277 0.079 35 0.21 278 0.087 39 0.64 279 0.092 40 0.037 280 0.092 45 0.062 281 0.093 46 0.13 282 0.094 47 >100 283 0.094 50 0.039 284 0.098 51 0.0052 285 0.10 52 0.056 286 0.10 53 0.043 287 0.10 54 0.063 288 0.10 55 0.038 289 0D1

- 255 044500

57 0.14 290 ο,ιι 58 0.028 291 ο,ιι 60 1.6 292 ο,ιι 61 0.056 293 ο,ιι 64 55 294 0.14 65 0.0055 295 0.14 66 0.048 296 0.15 67 0.0066 297 0.16 68 0.052 298 0.16 69 >100 299 0.16 70 0.0089 300 0.16 73 0.026 301 0.17 75 0.14 302 0.18 76 0.067 303 0.18 78 >100 304 0.19 79 0.067 305 0.21 85 θ,ι 306 0.21 89 2.8 307 0.21 90 0.63 308 0.21 95 1.1 310 0.22 96 0.36 312 0.24 98 0.13 313 0.25 99 0.0094 314 0.25 101 0.19 315 0.25 103 0.40 316 0.41 104 0.44 317 0.29 106 0.0024 318 0.29 115 3.8 319 0.30 116 0.17 320 0.30 117 0.77 321 0.31 118 0.018 322 0.32 122 0.011 323 0.34 125 0.023 324 0.35

- 256 044500

127 0.0064 326 0.38 128 0.10 327 0.43 130 0.019 328 0.44 138 >100 329 0.47 140 0.006 330 0.50 144 0.02 331 0.52 146 >100 332 0.53 147 0.30 333 0.54 149 0.064 334 0.54 150 0.51 335 0.54 151 0.88 337 0.55 152 0.15 338 0.58 153 0.71 339 0.65 154 0.88 340 0.69 155 1.6 341 0.73 156 >100 342 0.79 157 0.22 343 0.81 159 2.0 344 0.82 160 2.9 345 0.99 161 5.7 346 1.1 163 2.1 347 1.2 164 2.6 348 1.3 163 0.22 349 1.4 166 3.8 350 1.5 176 h,o 351 1.5 183 3.2 352 1.6 186 3.0 353 1.9 189 7.3 354 2.5 190 4.2 355 2.8 192 0.20 356 h,o 193 0.13 357 5.0 194 0.44 358 6.5 195 0.12 359 8.0

- 257 044500

196 2.1 360 9.6 198 0.50 362 >100 199 0.38 363 >100 200 1.6 364 >100 201 22 365 >100 202 0.40 366 >100 213 0.71 367 >100 214 0.55 368 >100 215 0.48 369 >100 216 0.064 370 >100 217 0.067 371 >100 218 0.093 372 >100 220 0.063 373 >100 221 0.33 375 >100 222 0.068 417 0.15 223 0.042 421 2.8 225 0.07 422 >100 226 0.084 423 >100 227 1.5 424 >100 228 oh and 425 >100 229 0.12 274 0.072 230 3.6 419 0.29 231 h,o 426 20 232 1.1 427 2.4

Table 3. IC ₅₀ values in the API-luciferase reporter assay

EXAMPLE No. API-luciferase reporter IR ₅ o (mKM) EXAMPLE No. AP 1-luciferase reporter IC50 (µM) AND 0.08 136 0.23 12 0.58 137 0.19 13 1.3 138 0.66

- 258 044500

15 1.82 139 0.13 17 0.35 141 0.24 19 0.83 145 0.57 20 0.4 146 1.87 22 1.53 148 0.87 25 1.15 156 10 28 4.45 158 4.59 32 0.36 162 4.81 36 1.6 188 10 37 2.83 211 0.52 43 0.4 212 1.14 44 0.35 362 8.37 47 2.55 363 10 48 2.16 364 1.57 69 0.51 365 2.22 78 0.14 372 0.14 FROM 0.58 373 0.26 114 4.12 374 7.93 120 1.3 376 10 121 0.34 379 7.84 123 0 381 0.19 124 ο,ι 384 0.18 126 0 385 0.06 129 0 386 0.05 131 0.5 389 0.65 132 0.08 390 0.8 133 0.28 391 0.07 134 0.6 422 3.49 135 0.34 423 1.88 136 0.23 424 10

- 259 044500

Table 4. IC ₅₀ values in the DGKAPl-luciferase reporter assay

EXAMPLE No. DGKAP1luciferase reporter IC50 (µM) EXAMPLE No. DGKAP1luciferase reporter IC50 (µM) 17 1.33 364 10 49 1.7 365 10 124 0.57 377 10 129 0.87 378 10 143 1.28 382 10 156 10 383 10 158 10 387A 10 162 10 387 3.33 177 10 388 3.33 224 10 415 0.42 315 10 - -

Table 5. Normalized EC50 activity values in the CD8 GLO assay

EXAMPLE No. CD8 GLO Normalized EC50 (µM) EXAMPLE No. CD8 GLO Normalized EC50 (µM) 32 0.18 377 10 49 0.17 378 1.74 124 0.02 382 1.79 129 0.06 383 0.06 141 0.01 387A 10 143 0.01 387 0.18 156 0.14 388 0.48 158 0.92 405 0.02 162 10 412 o, s 177 10 415 0.02 224 4.28 425 0.19 315 1.26 - -

- 260 044500

Table 6. Normalized EC50 activity values in the HuCD8 INFG assay

EXAMPLE No. HuCD8 INFG Normalized ECso (µM) EXAMPLE No. HuCD8 INFG Normalized EC50 (µM) 13 10 398V 5.97 17 0.52 398V 0.6 44 0.18 399 0.02 124 0.01 400 0.05 137 0.04 401 10 141 0.03 402 1.78 143 0.09 403 1.49 251 0.58 406 0.41 252 10 407 0.12 253 2.51 408 0 256 0.24 410 0.09 381 0.07 411 0.02 386 0.96 414 0.16 392 0.23 416 0.03 393 0.17 423 0.46 394 0.01 413 0.04 395 0.09 428 0.03 396 2.66 429 0.08 397 4.47 430 o, s 398 1.21 432 0.06

Table 7. IC ₅₀ activity values in the huCD8 pERK assay

EXAMPLE No. HuCD8 pERK IC50 (µM) 124 0.36 143 0.06

Table 8. EC50 Agonist Activity Values in Whole Blood INF-γ Assay

EXAMPLE No. INF-γ whole blood EC so (µM) agonists 17 0.67 124 0.14 145 1.33

- 261 044500

Table 9. Normalized values for EC ₅₀ agonist activity in whole blood INF-γ assay

EXAMPLE No. INF-γ whole blood Normalized EC so (µM) agonists EXAMPLE No. Whole Blood INF-γ Normalized ECso (µM) Agonists 13 5 372 5 17 1D5 386 1.24 44 0.9 391 3.5 121 0.55 406 2.19 124 0.1 407 0.78 136 1.16 412 3.01 137 5 423 10 145 1.87 425 10

Table 10. Normalized values for EC ₅₀ agonist activity in whole blood INF-γ assay

EXAMPLE No. msCTL INF-γ IC50 (µM) EXAMPLE No. msCTL INF-γ IC50 (µM) 13 0.64 378 10 17 0.19 381 0.7 36 10 383 10 44 0.13 385 4.09 48 10 386 0.59 121 0.05 389 5.05 124 0.04 391 0.73 133 0.06 392 1.85 134 1.14 399 0.41 136 0.09 404 9.84 137 0.23 406 0.16 141 0.29 407 0.04 143 1.41 412 0.36 145 0.46 415 10 177 10 423 0.7 224 10 424 10 362 10 425 2.98 372 2.55 428 0.59

In table Table 1 shows the IR ₅₀ activity values obtained in an in vitro DGK inhibition assay using DGKa- and DGKζ-containing liposomes (LIPGLO assay) and detergent/lipid micelles. Compounds of the present invention shown in examples 2-3, 6-37, 39-40, 43-55, 57-58, 60-61, 64-70, 73, 75-76, 78-79, 85, 89- 90, 95-96, 98-99, 101, 103-106, 114-115, 117-141, 143-157, 159-160, 162-166, 176-177, 183, 189-190, 192-200, 202, 211-218, 220-234, 239, 244, 250-256, 258-308, 310, 312-324, 326-335, 337-357, 359-360, 362-373, 375, 377-383, 385-417, 419 and 421-432 had an IC ₅₀ value of <240 μM in one or more of the DGKa and DGKZ LIPGLO assays and the DGKa and DGKZ assays, indicating inhibition of one or both of the DGKa and DGKZ enzymes.

The compounds of the present invention have inhibitory activity against one or both of the enzymes DGKa and DGKZ, and therefore can be used for the treatment of diseases associated with inhibition of the activity of DGKa and DGKZ.

Nucleotide sequence encoding hDGKa-(M1-S735)-Ct-TVMV-His:

- 262 044500

ATGGCCAAGG AGAGGGGCCT AATAAGCCCC AGTGATTTTG CCCAGCTGCA 51 AAAATACATG GAATACTCCA CCAAAAAGGT CAGTGATGTC CTAAAGCTCT 101 TCGAGGATGG CGAGATGGCT AAATATGTCC AAGGAGATGC CATTGGGTAC 151 GAGGGATTCC AGCAATTCCT GAAAATCTAT CTCGAAGTGG ATAATG TTCC 201 CAGACACCTA AGCCTGGCAC TGTTTCAATC CTTTGAGACT GGTCACTGCT 251 TAAATGAGAC AAATGTGACA AAAGATGTGG TGTGTCTCAA TGATGTTTCC 301 TGCTACTTTT CCCTTCTGGA GGGTGGTCGG CCAGAAGACA AGTTAGAATT 351 CACCTTCAAG CTGTACGACA CGGACAGAAA TGGGATC CTG GACAGCTCAG 401 AAGTGGACAA AATTATCCTA CAGATGATGC GAGTGGCTGA ATACCTGGAT 451 TGGGATGTGT CTGAGCTGAG GCCGATTCTT CAGGAGATGA TGAAAGAGAT 501 TGACTATGAT GGCAGTGGCT CTGTCTCTCA AGCTGATGG GTCCGGGCTG 551 GGGCCACCAC CGTGCCACTG CTAGTGCTGC TGGGTCTGGA GATGACTCTG 601 AAG GACGACG GACAGCACAT GTGGAGGCCC AAGAGGTTCC CCAGACCAGT 651 CTACTGCAAT CTGTGCGAGT CAAGCATTGG TCTTGGCAAA CAGGGACTGA 701 GCTGTAACCT CTGTAAGTAC ACTGTTCACG ACCAGTGTGC CATGAAAGCC 751 CTGCCTTGTG AAGTCAGCAC CTATGCCAAG TCTCGGAAGG ACATTGGTGT 801 CCAATCACAT GTGTGGGTGC GAGGAGGCTG TGAGTCCGGG CGCTGCGACC 851 GCTGTCAGAA AAAGATCCGG ATCTACCACA GTCTGACCGG GCTGCATTGT 901 GTATGGTGCC ACCTAGAGAT CCACGATGAC TGCCTGCAAG CGGTGGGCCA 951 TGAGTGTGAC TGTGGGCTGC TCCGGGATCA CATCCTGCCT CCATCTTCCA 1001 TCTATCCCAG TGTCCTGGCC TCTGGA CCGG ATCGTAAAAA TAGCAAAACA 1051 AGCCAGAAGA CCATGGATGA TTTAAATTTG AGCACCTCTG AGGCTCTGCG 1101 GATTGACCCT GTTCCTAACA CCCACCCACT TCTCGTCTTT GTCAATCCTA 1151 AGAGTGGCGG GAAGCAGGGG CAGAGGGTGC TCTGGAAGTT CCAGTATATA 1201 TTAAACCCTC GACAGGTGTT CAACCTCCTA AAGGATGGTC CTGAGATAGG 1251 GCTCCGATTA TTCAAGGATG TTCCTGATAG CCGGATTTTG GTGTGTGGTG 1301 GAGACGGCAC AGTAGGCTGG ATTCTAGAGA CCATTGACAA AGCTAACTTG 1351 CCAGTTTTGC CTCCTGTTGC TGTGTTGCCC CTGGGTACTG GAAATGATCT 1401 GGCTCGATGC CTAAGATGGG GAGGAGGTTA TGAAGGACA G AATCTGGCAA 1451 AGATCCTCAA GGATTTAGAG ATGAGTAAAG TGGTACATAT GGATCGATGG 1501 TCTGTGGAGG TGATACCTCA ACAAACTGAA GAAAAAGTG ACCCAGTCCC 1551 CTTTCAAATC ATCAATAACT ACTTCTCTAT TGGCGTGGAT GCCTCTATTG 1601 CTCATCGATT CCACATCATG CGAGA GAAAT ATCCGGAGAA GTTCAACAGC 1651 AGAATGAAGA ACAAGCTATG GTACTTCGAA TTTGCCACAT CTGAATCCAT 1701 CTTCTCAACA TGCAAAAAGC TGGAGGAGTC TTTGACAGTT GAGATCTGTG 1751 GGAAACCGCT GGATCTGAGC AACCTGTCCC TAGAAGGCAT CGCAGTGCTA 1801 AACATCCCTA GCATGCATGG TGGCTCCAAC CTCTGGGGTG ATACCAGGAG 1851 ACC CCATGGG GATATCTATG GGATCAACCA GGCCTTAGGT GCTACAGCTA 1901 AAGTCATCAC CGACCCTGAT ATCCTGAAAA CCTGTGTACC AGACCTAAGT 1951 GACAAGAGAC TGGAAGTGGT TGGGCTGGAG GGTGCAATTG AGATGGGCCA 2001 AATCTATACC AAGCTCAAGA ATGCTGGACG TCGGCTGGCC AAGTGCTCTG 2051 AGATCACCTT CCACACCACA AAAACCCTTC CCATGCAAAT TGACGGAGAA 2101 CCCTGGATGC AGACGCCCTG TACAATCAAG ATCACCCACA AGAACCAGAT

- 263 044500

2151

2201

2251

GCCCATGCTC ATGGGCCCAC CCCCCCGCTC CACCAATTTC TTTGGCTTCT TGAGCGGATC CTCGGAGACA GTGCGGTTTC AGGGACACCA CCACCATCAC CACTGA (SEQ Ш NO: 1)

Amino acid sequence of hDGKa-(Ml-S735)-Ct-TVMV-His:

0001

0061

0121

0181

0241

0301

0361

0421

0481

0541

0601

0661

0721

MAKERGLISP LEVDNVPRHL LYDTDRNGIL VRAGATTVPL TVHDQCAMKA VWCHLEIHDD STSEALRIDP FKDVPDSRIL NLAKILKDLE REKYPEKFNS NIPSMHGGSN GAIEMGQIYT MGPPPRSTNF

SDFAQLQKYM SLALFQSFET DSSEVDKIIL LVLLGLEMTL LPCEVSTYAK CLQAVGHECD VPNTHPLLVF VCGGDGTVGW MSKWHMDRW RMKNKLWYFE LWGDTRRPHG KLKNAGRRLA FGFLSGSSET

EYSTKKVSDV GHCLNETNVT QMMRVAEYLD KDDGQHMWRP SRKDIGVQSH CGLLRDHILP VNPKSGGKQG ILETIDKANL SVEVIPQQTE FATSESIFST DIYGINQALG KCSEITFHTT VRFQGHHHHH

LKLFEDGEMA KDWCLNDVS WDVSELRPIL KRFPRPVYCN VWVRGGCESG PSSIYPSVLA QRVLWKFQYI PVLPPVAVLP EKSDPVPFQI CKKLEESLTV ATAKVITDPD KTLPMQIDGE H 0751

KYVQGDAIGY CYFSLLEGGR QEMMKEIDYD LCESSIGLGK RCDRCQKKIR SGPDRKNSKT LNPRQVFNLL LGTGNDLARC INNYFSIGVD EICGKPLDLS ILKTCVPDLS PWMQTPCTIK

EGFQQFLKIY PEDKLEFTFK GSGSVSQAEW QGLSCNLCKY IYHSLTGLHC SQKTMDDLNL KDGPEIGLRL LRWGGGYEGQ ASIAHRFHIM NLSLEGIAVL DKRLEWGLE ITHKNQMPML

0060

0120

0180

0240

0300

0360

0420

0480

0540

0600

0660

0720 (SEQ Ш NO: 2)

Nucleotide sequence encoding hDGKY(Ml-A928)-transcript variant-2 Ct-TVMV-His:

101

151

201

251

301

351

401

451

501

551

601

651

701

751

801

851

901

951

1001

1051

1101

1151

1201

1251

1301

1351

1401

ATGGAGCCGC GGCTTCCGCC ACAAGGCGCC TTCGGGCACA CCCTCCGCCC GGAGTACAGT TTCGAGACCA TGTAGCCAGG GCAAGATTGT TTCCGCTGTA GCCAACCTTT AGTGTCGGCA AAGGAGATTG CAAGGTGTCC GGGTCCACGC AGGCCCCAGA CAAGAGGAAG TCATCATGAG GTGAACCCCA CCTCTGGTAT CCAAGGAGGC GCG TGCGGGG GCTACGCCTG GCAACGACTT CCTGTGTCCA GGACCGCTGG ACCGAGATGA TACTTCAGCC TCGAGAGGCC

GGGACGGTAG TCGTCCAGCG GCGGCGACTC GGAAAGCCAT ACCCTGGGG GGACTGGAGC ACGTGTCCGG ATGCTGCAGA GGTGCACACG AGCCGTCCTT GTACGGCACC CTGTGGGAAG TGGCCATCAG TGCTTCATGC AGCCGTGGTC ATACTCTGAA TCCAGCAAGA GCCCACCCCC AGAGTGGGGG CTCAATCCCC GCTGGAGATG GC GACGGCAC AAGCCGCCAC GGCCCGAACC AGATCCTCTC GACCTCCACG AGGCGCCACC TGGGCTTTGA AACCCAGAGA

CCCCGAGGCC GCTCCGAGCG AACAAGCGGC CACGAAGTCG CCCCGTGCAG GAGTCAGCGA GGACTTCTGC AGTCAGTGTC CCCTGCATCG CCGTGAATCA ACTGGGTACA GGATTCCAGC CTGCTCGTGG TGCAGCAGAT ATCCCGCCCA AGCAAGCAAG AAGGGCCTGA TCCCCGCTCA CAACCAGGT GACAAGTCTT TACCGCAAAG GGTGGGCTGG CCCCTGTTGC CTCAACTGGG CCACGTGGAG CTGAGCCCAA GACCGGTTGC CGCCCACGTC AATTCAACAG

CGGAGCAGCG CGACGCCGGT GCTTCCCGGG GGCCTCCAGC CGAGTCAGAG CATATGGGGA TACGTTGGGG TCGAAGAAAG AGCAGCTGGA GGCTCCAGGA CAGACGACGC AGAAGTTCAC TGCAAGCAGG CGAGGAGCCG CCTGGATCCT AAGAAGAAGA GGAGGGCCGC TGAAGCCCCT GCAAAGATCA CGACCTGAGC TGCACAAC CT ATCCTCTCCA CATCCTGCCC GTGGGGGCTA GAGGGGAACG CCCCGAGGCA CCCTGGATGT ACCCTGGAGT CCGCTTTCGG

ACTCCGAGTC CCCGAGCCGG GCTGCGGCTC ACCTGGCCCC CGGCAGATCC GCACATCTGG AGCAGTACTG TGCGCAGCCT GAAGATAAAT ATGTCCGCGA CAGGACGGCA CTTCCACAGC CATACCACAG TGCTCGCTGG CCGCGCCCGG GGGCATCCTT TGGAGACCCT GCTGGTGTTT TCCAGTCTTT CAGGGAGGGC GCGGATCCTG CCCTGGACCA CTGGGTACTG CACAGATGAG TGGTACAGCT GGGCCTGAGG CTTCAACAAC TCCACGAGTC AATAAGATGT

- 264 044500

1451 TCTACGCCGG GACAGCTTTC TCTGACTTCC TGATGGGCAG CTCCAAGGAC 1501 CTGGCCAAGC ACATCCGAGT GGTGTGTGAT GGAATGGACT TGACTCCCAA 1551 GATCCAGGAC CTGAAACCCC AGTGTGTTGT TTTCCTGAAC ATCCCCAGGT 1601 ACTGTGCGGG CACCATGCCC TGGG GCCACC CTGGGGAGCA CCACGACTTT 1651 GAGCCCCAGC GGCATGACGA CGGCTACCTC GAGGTCATTG GCTTCACCAT 1701 GACGTCGTTG GCGCGCCTGC AGGTGGGCGG ACACGGCGAG CGGCTGACGC 1751 AGTGTCGCGA GGTGGTGCTC ACCACATCCA AGGCCATCCC GGTGCAGGTG 1801 GATGGCGAGC CCTGCAAGCT TGCAGCCTCA CGCATCCGCA TCGCCCTGCG 1851 CAACCAGGCC ACCATGGTGC AGAAGGCCAA GCGGCGGAGC GCCGCCCCCC 1901 TGCACAGCGA CCAGCAGCCG GTGCCAGAGC AGTTGCGCAT CCAGGTGAGT 1951 CGCGTCAGCA TGCACGACTA TGAGGCCCTG CACTACGACA AGGAGCAGCT 2001 CAAGGAGGCC TCTGTGCCGC TGGGCACTGT GGTGGTCCCA GGAGA CAGTG 2051 ACCTAGAGCT CTGCCGTGCC CACATTGAGA GACTCCAGCA GGAGCCCGAT 2101 GGTGCTGGAG CCAAGTCCCCC GACATGCCAG AAACTGTCCC CCAAGTGGTG 2151 CTTCCTGGAC GCCACCACTG CCAGCCGCTT CTACAGGATC GACCGAGCCC 22 01 AGGAGCAGCT CAACTATGTG ACT GAGATCG CACAGGATGA GATTTATATC 2251 CTGGACCCTG AGCTGCTGGG GGCATCGGCC CGGCCTGACC TCCCAACCCC 2301 CACTTCCCCT CTCCCCACCT CACCCTGCTC ACCCACGCCC CGGTCACTGC 2351 AAGGGGATGC TGCACCCCCT CAAGGTGAAG AGCTGATTGA GGCTGCCAAG 2401 AGGAACGACT TCTGTAAGCT CCAGGAGCTG CACCGAGCTG GGGGCGACCT 2 4 5 1 CATGCACCGA GACGAGCAGA GTCGCACGCT CCTGCACCAC GCAGTCAGCA 2501 CTGGCAGCAA GGATGTGGTC CGCTACCTGC TGGACCACGC CCCCCCAGAG 2551 ATCCTTGATG CGGTGGAGGA AAACGGGGAG ACCTGTTTGC ACCAAGCAGC 2601 GGCCCTGGGC CAGCGCACCA TCTGCCACTA CATCGTGGAG GCCGGGGCCT 2 651 CGCTCATGAA GACAGACCAG CAGGGCGACA CTCCCCGGCA GCGGGCTGAG 2701 AAGGCTCAGG ACACCGAGCT GGCCGCCTAC CTGGAGAACC GGCAGCACTA 2751 CCAGATGATC CAGCGGGAGG ACCAGGAGAC GGCTGTGGGA TCCTCGGAGA 2801 CAGTGCGGTT TCAGGGACAC CACCACCATC ACCACTGA (SEQIDNO: 3 )

Amino acid sequence hDGK^-(Ml-A928)-transcript variant-2 CtTVMV-His:

0001 MEPRDGSPEA RSSDSESASA SSSGSERDAG PEPDKAPRRL NKRRFPGLRL FGHRKAITKS 0060 0061 GLQHLAPPPP TPGAPCSESE RQIRSTVDWS ESATYGEHIW FETNVSGDFC YVGEQYCVAR 0120 0121 MLQKSVSRRK CAACKIWHT PCIEQLEKIN FRCK PSFRES GSRNVREPTF VRHHWVHRRR 0180 0181 QDGKCRHCGK GFQQKFTFHS KEIVAISCSW CKQAYHSKVS CFMLQQIEEP CSLGVHAAW 0240 0241 IPPTWILRAR RPQNTLKASK KKKRASFKRK SSKKGPEEGR WRPFIIRPTP SPLMKPLLVF 03 00 0301 VNPKSGGNQG AKIIQSFLWY LNPRQVFDLS QGGPKEALEM YRKVHNLRIL ACGGDGTVGW 0360 0361 ILSTLDQLRL KPPPPVAILP LGTGNDLART LNWGGGYTDE PVSKILSHVE EGNWQLDRW 0420 0421 DLHAEPNPEA GPEDRDEGAT DRLPLDVFNN YFSLGFDAHV TLEFHESREA NPEKFNSRFR 0480 0481 NKMFYAGTAF SDFLMGSSKD LAKHIRWCD GMDLTPKIQD LKP QCWFLN IPRYCAGTMP 0540 0541 WGHPGEHHDF EPQRHDDGYL EVIGFTMTSL AALQVGGHGE RLTQCREWL TTSKAIPVQV 0600 0601 DGEPCKLAAS RIRIALRNQA TMVQKAKRRS AAPLHSDQQP VPEQLRIQVS RVSMHDYEAL 0660 066 1 HYDKEQLKEA SVPLGTVWP GDSDLELCRA HIERLQQEPD GAGAKSPTCQ KLSPKWCFLD 0720 0721 ATTASRFYRI DRAQEHLNYV TEIAQDEIYI LDPELLGASA RPDLPTPTSP LPTSPCSPTP 0780 0781 RSLQGDAAPP QGEELIEAAK RNDFCKLQEL HRAGGDLMHR DEQSRTLLHH AVSTGSKDW 0840 0841 RYLLDHAPPE ILDAVEENGE TCLHQAAALG QRTICHYIVE AGASLMKTDQ QGDTPRQRAE 0900 0 901 KAQDTELAAY LENRQHYQMI QREDQETAVG SSETVRFQGH HHHHH 0945 (SEQ ID NO: 4)

Claims (10)

CLAIM 1. Compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein R1 represents H, Cl, Br, -CN, C1. ₂ alkyl, -CH= _CH2 , _-OSH3 , -C(O)OH, -C(O)OSH3, -C( _O )N( _CH3 )2, -NH2 or -NHC(O)OC( CH3)3; R2 represents -CH3, -CH2CH3, -CH2CH2CH3, -CH2CH2CH2CH3, -CH2CN, -CH2CH2CH2CN, -CH2CH2CF3, -CH ₂ CH=CH ₂ , -CH ₂ CH ₂ CH=CF ₂ , -sns sn, -CH ₂ CH ₂ OCH ₃ , -CH ₂ CH ₂ CH ₂ OCH ₃ , -CH2CH2CH2OCH2CH2OCH3, -CH2CH2CH2C(O)CH3, -CH2C(O)OCH2CH3, -CH2(cyclopropyl) or -CH2CH2(dioxanyl); - 265 044500 R ₃ represents H, F, Cl, Br, -CN, -CH ₃ , -CF ₃ , -NO2, -C(O)OCH2CH ₃ or -C(O)CF ₃ ; R4 represents: (a) 2,3-dihydro-1H-indenyl having from 1 to 2 substituents independently selected from F, -OH, -OSH ₃ and -OSH ₂ CH=CH ₂ ; or (b) -CH ₂ R _y , -C(CH ₃ ) ₂ R _y , -CHR _x R _y , -CH ₂ CH(OH)R _x , -CH(CH ₃ )(CH ₂ CH ₂ OCH ₃ ) or cyclopropyl having a fluorophenyl substituent; R _x represents C _1-2 alkyl, -CH(CH ₃ ) ₂ , -C(CH ₃ ) ₃ , -CH ₂ C(CH ₃ ) ₃ , -CH2OH, -CH2NH2, cyclopropyl, cyclobutyl, cyclohexyl or phenyl, having from 0 to 2 substituents independently selected from F, Cl, -OH and -OCH3; And R _y is 1,3-benzodiazolyl, indazolyl, indolyl, ethyl indolyl, indolinyl, naphthalenyl, hydroxynaphthalenyl, oxoindolinyl, pyridinyl, methoxypyridinyl, pyrimidinyl or phenyl, having from 0 to 3 substituents independently selected from F, Cl, Br, - OH, -CN, -CH3, -C(CH3)3, -CHF2, -CF ₃ , -OCH ₃ , -OCF ₃ , -OCH ₂ CH=CH ₂ , -OCH2C CH, -OCH ₂ (cyanopyridinyl), - NH ₂ , -NHS(O) ₂ CH ₃ , -N(CH ₃ )(CH ₂ CH ₃ ), -NHC(O)CH3, -NHC(O)O(C(CH3)3), -NHC(O )(phenyl), -NHC(O)NH(phenyl) and phenyl. 2. A compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein R4 is -CH2Ry, -CHRxRy or -CH2CH(OH)Rx. 3. The compound according to claim 2, or a pharmaceutically acceptable salt thereof, in which R _x represents phenyl having from 0 to 2 substituents independently selected from F, Cl, -OH, C1-2 alkyl, -CHF2, -OCH3, -OCH ₂ CH=CH ₂ and -OCH2C CH; And R _y represents phenyl having from 0 to 3 substituents independently selected from F, Cl, Br, -OH, -CN, C _1-4 alkyl, C _1-2 fluoroalkyl, C _1-2 alkoxy, C _1-2 fluoroalkoxy, -OSH ₂ CH=CH ₂ , -OSH ₂ C=CH, -OSH ₂ (cyanopyridinyl), -NR _c R _c , -NHS(O) ₂ CH ₃ , -NHC(O)(C _1-2 alkyl ), -NHC(O)O(C _1-4 alkyl), -NHC(O)(phenyl), -NHC(O)NH(phenyl) and phenyl. 4. A compound according to claim 1, or a pharmaceutically acceptable salt thereof, in which R4 is -CHRxRy; Rx is phenyl having from 0 to 2 substituents independently selected from F, Cl, -OH and -OCH3; And Ry is phenyl having from 0 to 3 substituents independently selected from F, Cl, Br, -OH, -CN, _-CH3 , -C( _CH3 ) ₃ , -CHF2, _-CF3 , _-OCH3 , -OCF ₃ , -OCH ₂ CH=CH ₂ , -OCH2C CH, -OCH ₂ (cyanopyridinyl), -NH2, -NHS(O)2CH3, -N(CH ₃ )(CH ₂ CH ₃ ), -NHC(O )CH ₃ , -NHC(O)O(C(CH ₃ ) ₃ ), -NHC(O)(phenyl), -NHC(O)NH(phenyl) and phenyl. 5. A compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein R4 is 2,3-dihydro-1H-indenyl having from 0 to 2 substituents independently selected from F, Cl, -OH, C _1-2 alkyl, C _1-2 fluoroalkyl, C _1-2 alkoxy and -OSH ₂ CH=CH ₂ . 6. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, which is: ethyl 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-6-bromo-1-methyl-2- oxo-1,2-dihydro-1,5-naphthyridine-3-carboxylate (1); 6-bromo-4-(4-(2-hydroxybenzyl)piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydro1,5-naphthyridine-3-carbonitrile (2); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-1-methyl-3-nitro1,5-naphthyridin-2(1H)-one (3); 6-bromo-4-{4-[(4-fluorophenyl)[2-(prop-2-yn-1-yloxy)phenyl]methyl]piperazin-1 -yl}-1 -methyl-2-oxo-1, 2-dihydro-1,5-naphthyridine-3-carbonitrile (5); 6-bromo-4-{4-[(4-fluorophenyl)[2-(prop-2-yn-1 -yloxy)phenyl]methyl]piperazin-1 -yl}-1 -methyl-2-oxo- 1,2-dihydro-1,5-naphthyridine-3 carbonitrile (5-7); 6-bromo-4-{4-[(4-fluorophenyl)(2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo1,2-dihydro-1,5-naphthyridine-3-carbonitrile (8-10); 8-{4-[(4-fluorophenyl)(2-hydroxyphenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2,7-dicarbonitrile (eleven); 6-bromo-4-{4-[(4fluorophenyl)(2-methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3 carbonitrile ( 12-14); 6-bromo-4-{4-[(4-fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl]piperazin-1-yl}-1-methyl2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile (15-17); 8-{4-[(4-fluorophenyl)(2-methoxyphenyl)methyl]piperazin-1-yl}-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridin-2 -carbonitrile (18-20); 6-bromo4-[4-(6-methoxy-2,3-dihydro-1H-inden-1 -yl)piperazin-1 -yl]-1 -methyl-2-oxo-1,2-dihydro-1,5 -naphthyridine3-carbonitrile (21); 6-bromo-4-{4-[1-(4-fluorophenyl)ethyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile (22- 24); 6-bromo-4-{4-[1-(4-fluorophenyl)propyl]piperazin-1-yl}-1-methyl-2-oxo1,2-dihydro-1,5-naphthyridine-3-carbonitrile (25- 27); 6-bromo-4-{4-[2-(4-fluorophenyl)-2-hydroxyethyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3 -carbonitrile (28-30); 6-bromo-4-{4-[1(4-fluorophenyl)-2-hydroxyethyl]piperazin-1 -yl} -1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3 - carbonitrile (31); 8-{4-[1-(4-fluorophenyl)propyl]piperazin-1-yl}-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile ( 32-34); 6-bromo-4-{4-[cyclopropyl(4-fluorophenyl)methyl]piperazin-1-yl}-1-methyl2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (35); 8-{4-[2-(4-fluorophenyl)-2-hydroxyethyl]piperazin-1-yl}-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridin-2 -carbonitrile (36); 8-{4-[1-(4-fluorophenyl)-2-hydroxyethyl]piperazin-1-yl}-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridin-2 -carbonitrile (37); 1 -methyl-4-{4-[(naphthalene-1 -yl)methyl]piperazin-1 -yl}-3-nitro-1,2-dihydro-1,5-naphthyridin-2-one (38); 6-chloro-4-{4-[(4-fluorophenyl)[2-(prop-2-yn-1-yloxy)phenyl]methyl]piperazin-1-yl}-1-methyl-2-oxo- 266 044500 1,2-dihydro-1,5-naphthyridine-3-carbonitrile (39); 8-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-5-methyl7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (40); 8-(4-benzhydrylpiperazin-1-yl)-5methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridin-2-carbonitrile (41); 8-(4-((2-hydroxyphenyl)(phenyl)methyl)piperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (42); 8(4-(1-(4-fluorophenyl)ethyl)piperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridin-2-carbonitrile (43 -45); 8-(4-(6-methoxy-2,3-dihydro-III-inden-1-yl)piperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6dihydro-1,5 -naphthyridine-2-carbonitrile (46); 8-(4-(2-hydroxy-1-phenylethyl)piperazin-1-yl)-5-methyl-7nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (47); 8-(4-(2-hydroxy-2-phenylethyl)piperazin1-yl)-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (48); 8-(4-(cyclopropyl(4fluorophenyl)methyl)piperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (49-51 ); 4-(4-benzhydrylpiperazin-1-yl)-6-bromo-1-methyl-3-nitro-1,5-naphthyridin-2(1H)-one (52); 4-(4(bis(4-fluorophenyl)methyl)piperazin-1-yl)-6-bromo-1-methyl-3-nitro-1,5-naphthyridin-2(1H)-one (53); 6bromo-4-(4-(( 1 -ethyl-1 N-indol-4-yl)methyl)piperazin-1 -yl)-1 -methyl-3-nitro-1,5-naphthyridin-2(1 H) -on (54); 6-bromo-1-methyl-4-(4-(naphthalene-1 -ylmethyl)piperazin-1 -yl)-3-nitro-1,5-naphthyridin-2(1 H)-one (55); 6-bromo-4-(4-((4-fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl)piperazin-1-yl)-1-methyl-3-nitro-1,5-naphthyridine-2(1 N)-on (56-58); 6-bromo-4-(4-((4-fluorophenyl)(2-methoxy-6-methylphenyl)methyl)piperazin-1 -yl)-1 methyl-3-nitro-1,5-naphthyridine-2(1H) -on (59-61); tert-butyl (8-(4-(bis(4-fluorophenyl)methyl)piperazin-1yl)-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridin-2-yl )carbamate (62); 6-amino-4-(4-(bis(4-fluorophenyl)methyl)piperazin-1 -yl)-1 -methyl-3-nitro-1,5-naphthyridin-2(1 H)-one (63); 6-bromo-4-(4-(2-(difluoromethyl)benzyl)piperazin-1-yl)-2-oxo-1 -(prop-2-in-1 -yl)-1,2-dihydro-1, 5-naphthyridine-3-carbonitrile (64); 6-bromo-4-(4-(2-hydroxybenzyl)piperazin-1-yl)-2-oxo-1-(prop-2-in-1-yl)-1,2-dihydro-1,5-naphthyridine3 -carbonitrile (65); 6-bromo-4-(4-(2-hydroxy-4-methylbenzyl)piperazin-1 -yl)-2-oxo-1 -(prop-2-in-1 -yl)1,2-dihydro-1, 5-naphthyridine-3-carbonitrile (66); 6-bromo-4-(4-(4-fluoro-2-hydroxybenzyl)piperazin-1yl)-2-oxo-1 -(prop-2-in-1 -yl)-1,2-dihydro-1,5 -naphthyridine-3-carbonitrile (67); 6-bromo-4-(4-(4-fluoro-2methoxybenzyl)piperazin-1 -yl)-2-oxo-1 -(prop-2-in-1 -yl)-1,2-dihydro-1,5 -naphthyridine-3-carbonitrile (68); 6-bromo-4-(4-(2-hydroxy-4,6-dimethylbenzyl)piperazin-1 -yl)-2-oxo-1-(prop-2-in-1 -yl)-1,2-dihydro -1,5-naphthyridine-3-carbonitrile (69); 6-bromo-4-(4-((4-fluorophenyl)(2-hydroxyphenyl)methyl)piperazin1-yl)-1-methyl-3-nitro-1,5-naphthyridin-2(1H)-one (70) ; 6-bromo-4-(4-((2-fluoro-4-methylphenyl)(2-hydroxyphenyl)methyl)piperazin-1 -yl)-1 -methyl-3-nitro-1,5-naphthyridin-2(1 N)-on (71); 6-bromo-4-(4-((2,4-dimethylphenyl)(2-hydroxyphenyl)methyl)piperazin-1-yl)-1-methyl-3-nitro-1,5-naphthyridine-2(1H)- he (72); 6bromo-4-(4-((2-hydroxyphenyl)(ortho-tolyl)methyl)piperazin-1-yl)-1-methyl-3-nitro-1,5-naphthyridin2(1H)-one (73); 6-bromo-4-(4-((3-fluoro-2-hydroxyphenyl)(4-fluorophenyl)methyl)piperazin-1-yl)-1-methyl-3nitro-1,5-naphthyridine-2(1H)- he (74-76); 6-bromo-4-(4-((2-hydroxyphenyl)(phenyl)methyl)piperazin-1-yl)1-methyl-3-nitro-1,5-naphthyridin-2(1H)-one (77-79 ); 6-bromo-4-(4-((4-fluoro-2-hydroxyphenyl)(4fluorophenyl)methyl)piperazin-1-yl)-1-methyl-3-nitro-1,5-naphthyridine-2(1H)- he (80-82); 6-bromo-4-(4-((4fluorophenyl)(2-hydroxy-3-methylphenyl)methyl)piperazin-1-yl)-1-methyl-3-nitro-1,5-naphthyridin-2(1H)one (83-85); 6-bromo-4-(4-((4-fluorophenyl)(2-hydroxy-5-methylphenyl)methyl)piperazin-1-yl)-1-methyl-3nitro-1,5-naphthyridine-2(1H)- he (86); 6-bromo-4-(4-((4-fluorophenyl)(2-hydroxy-6-methylphenyl)methyl)piperazin-1-yl)-1-methyl-3-nitro-1,5-naphthyridine-2(1H )-on (87); 5-((2-((4-(6-bromo-1-methyl-3-nitro2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)piperazin-1 -yl)(4- fluorophenyl)methyl)-3-methylphenoxy)methyl)nicotinonitrile (88-90); 5-((2-((4-(6-bromo-1-methyl-3-nitro-2-oxo-1,2-dihydro-1,5-naphthyridin-4yl)piperazin-1-yl)(4- fluorophenyl)methyl)-5-fluorophenoxy)methyl)nicotinonitrile (91-93); 4-(4-((2(allyloxy)-6-methylphenyl)(4-fluorophenyl)methyl)piperazin-1-yl)-6-bromo-1-methyl-3-nitro-1,5-naphthyridine-2( 1H)-on (94-96); 8-(4-((4-fluorophenyl)(2-hydroxyphenyl)methyl)piperazin-1-yl)-5-methyl-7-nitro-6oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (97-99); 8-(4-((4-fluorophenyl) (2-(prop-2-yn-1yloxy)phenyl)methyl)piperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6- dihydro-1,5-naphthyridine-2-carbonitrile (100-102); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-6-methoxy-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile (103); 4-(4-((4-fluorophenyl)(2-hydroxyphenyl)methyl)piperazin-1-yl)-6methoxy-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (104); 6-bromo-4-(4-((4-fluorophenyl)(2-hydroxyphenyl)methyl)piperazin-1 -yl)-2-oxo-1-(prop-2-yn-1 -yl)-1,2 -dihydro-1,5-naphthyridine-3carbonitrile (105-107); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-6-bromo-1-(2-methoxyethyl)-2-oxo1,2-dihydro-1,5-naphthyridine-3-carbonitrile (113); 6-bromo-4-(4-((4-fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl)piperazin-1-yl)-1-(2-methoxyethyl)-2-oxo-1,2-dihydro -1,5-naphthyridine-3-carbonitrile (114116); 6-bromo-4-(4-((4-fluorophenyl)(2-hydroxyphenyl)methyl)piperazin-1-yl)-1-(2-methoxyethyl)-2-oxo-1,2dihydro-1,5-naphthyridine -3-carbonitrile (117); 8-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-7-nitro-6oxo-5-(prop-2-in-1-yl)-5,6-dihydro-1,5 -naphthyridine-2-carbonitrile (118); 8-(4-((4-fluoro-2hydroxyphenyl)(4-fluorophenyl)methyl)piperazin-1 -yl)-7-nitro-6-oxo-5 -(prop-2-in-1 -yl)-5 ,6-dihydro1,5-naphthyridine-2-carbonitrile (119-121); 8-(4-((4-fluoro-2-hydroxyphenyl)(4-fluorophenyl)methyl)piperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5 -naphthyridine-2-carbonitrile (122); 8-(4-((4-fluoro-2methoxyphenyl)(4-fluorophenyl)methyl)piperazin-1-yl)-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine -2-carbonitrile (123-125); 8-(4-((4-fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl)piperazin-1-yl)-7nitro-6-oxo-5-(prop-2-in-1-yl)-5 ,6-dihydro-1,5-naphthyridine-2-carbonitrile (126-128); 8-(4-((4-fluorine - 267 044500 phenyl)(2-methoxyphenyl)methyl)piperazin-1-yl)-7-nitro-6-oxo-5-(prop-2-in-1-yl)-5,6-dihydro-1,5 -naphthyridine-2-carbonitrile (129-131); 8-(4-((4-fluorophenyl)(2-hydroxyphenyl)methyl)piperazin-1-yl)-7nitro-6-oxo-5-(prop-2-yn-1-yl)-5,6-dihydro -1,5-naphthyridine-2-carbonitrile (132-134); 8-(4-(1-(4-fluorophenyl)ethyl)piperazin-1-yl)-7-nitro-6-oxo-5-(prop-2-in-1-yl)-5,6-dihydro- 1,5-naphthyridine-2-carbonitrile (135-137); 8-(4-(6-methoxy-2,3-dihydro-III-inden-1-yl)piperazin-1-yl)-7-nitro-6-oxo-5-(prop-2-in1-yl) -5,6-dihydro-1,5-naphthyridine-2-carbonitrile (138); 8-{4-[1-(4-fluorophenyl)propyl]piperazin-1-yl}7-nitro-6-oxo-5-(prop-2-yn-1-yl)-5,6-dihydro-1 ,5-naphthyridine-2-carbonitrile (139-141); 8-(4-(cyclopropyl(4-fluorophenyl)methyl)piperazin-1-yl)-7-nitro-6-oxo-5-(prop-2-in-1-yl)-5,6-dihydro-1 ,5-naphthyridine-2-carbonitrile (142-144); 8-(4-((4-fluorophenyl)(2-hydroxyphenyl)methyl)piperazin-1-yl)-5-(2methoxyethyl)-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine -2-carbonitrile (145); 8-(4-(6-methoxy-2,3dihydro-1H-inden-1 -yl)piperazin-1 -yl)-5-(2-methoxyethyl)-7-nitro-6-oxo-5,6-dihydro -1,5-naphthyridine2-carbonitrile (146); 8-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-5-(2-methoxyethyl)-7-nitro-6-oxo5,6-dihydro-1,5-naphthyridine-2-carbonitrile (147); 8-(4-(1-(4-fluorophenyl)ethyl)piperazin-1-yl)-5-(2methoxyethyl)-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2- carbonitrile (148-150); 5-(cyanomethyl)-8(4-(1-(4-fluorophenyl)ethyl)piperazin-1-yl)-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carbonitrile (151); 8-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-5-(cyanomethyl)-7-nitro-6-oxo-5,6-dihydro-1,5naphthyridin-2-carbonitrile (152 ); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-6-bromo-1-(cyanomethyl)2-oxo-1,2-dihydro-1,5-naphthyridin-3-carbonitrile ( 153); 6-bromo-1-(cyanomethyl)-4-(4-(1-(4-fluorophenyl)ethyl)piperazin-1-yl)-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (154); 6-bromo-1-(cyanomethyl)-4-(4-(1-(4-fluorophenyl)propyl)piperazin-1-yl)-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (155); 6-bromo-1 -(cyanomethyl)-4-(4-(6-methoxy-2,3-dihydro-1H-inden-1 -yl)piperazin-1 -yl)-2-oxo-1,2dihydro-1 ,5-naphthyridine-3-carbonitrile (156); 6-bromo-1-(cyanomethyl)-4-(4-((4-fluorophenyl)(2-hydroxyphenyl)methyl)piperazin-1 -yl)-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile (157); 6-bromo-1 (cyclopropylmethyl)-4-(4-((4-fluorophenyl)(2-methoxyphenyl)methyl)piperazin-1-yl)-2-oxo-1,2-dihydro1,5-naphthyridin-3- carbonitrile (158-160); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-6-bromo-1 (cyclopropylmethyl)-2-oxo-1,2-dihydro-1,5-naphthyridin-3-carbonitrile ( 161); 6-bromo-1 -(cyclopropylmethyl)-4-(4-(1 -(4-fluorophenyl)ethyl)piperazin-1 -yl)-2-oxo-1,2-dihydro-1,5-naphthyridin-3 - carbonitrile (162-164); 4-(4-benzhydrylpiperazin-1-yl)-3-nitro-1-(prop-2-yn-1-yl)-1,5-naphthyridin-2(1H)-one (165); 1-(2-(1,3-dioxan-2-yl)ethyl)-4-(4-benzhydrylpiperazin-1 -yl)-3-nitro-1,5-naphthyridin-2(1H)-one (166 ); 1allyl-4-(4-benzhydrylpiperazin-1-yl)-3-nitro-1,5-naphthyridin-2(III)-one (176); 4-(4-benzhydrylpiperazin-1 -yl)-1-butyl-3-nitro-1,5-naphthyridin-2(1 H)-one (177); 4-(4-(4-benzhydrylpiperazin-1 -yl)-3-nitro-2-oxo-1,5-naphthyridin-1(2H)-yl)butanenitrile (183); 4-(4-benzhydrylpiperazin-1-yl)-3-nitro-1(3,3,3-trifluoropropyl)-1,5-naphthyridin-2(1H)-one (185); 4-(4-benzhydrylpiperazin-1-yl)-1-(4,4-difluorobut-3-en-1-yl)-3-nitro-1,5-naphthyridin-2(1H)-one (186) ; 4-(4-benzhydrylpiperazin-1 -yl)-3-nitro-1-(4-oxopentyl)-1,5-naphthyridin-2(1H)-one (187); 4-(4-benzhydrylpiperazin-1-yl)-1-(3-(2-methoxyethoxy)propyl)-3-nitro-1,5-naphthyridin-2(1H)-one (189); 4-(4-benzhydrylpiperazin-1 -yl)-1 -(3-methoxypropyl)-3 nitro-1,5-naphthyridin-2(1 H)-one (190); 4-(4-benzhydrylpiperazin-1 -yl)-6-bromo-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3-carbonitrile (191); 6-bromo-4-(4-(4-fluoro-2-hydroxybenzyl)piperazin-1 -yl)-1 methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (192) ; 6-chloro-4-(4-(4-fluoro-2-hydroxybenzyl)piperazin-1 -yl)-1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (193 ); 4-(4-(4-fluoro-2hydroxybenzyl)piperazin-1 -yl)-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (194); 4-(4-(1-(2-(allyloxy)-4-fluorophenyl)ethyl)piperazin-1-yl)-6-chloro-1-methyl-2-oxo-1,2-dihydro-1,5- naphthyridine-3-carbonitrile (195); 4-(4-benzhydrylpiperazin-1 -yl)-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridin-3-carbonitrile (196); 6-chloro-4-(4-(2-hydroxybenzyl)piperazin-1 -yl)-1 -methyl-2-oxo-1,2dihydro-1,5-naphthyridine-3-carbonitrile (198); 6-chloro-4-(4-(3-(ethyl(methyl)amino)benzyl)piperazin-1yl)-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 199); 4-(4-(7-(allyloxy)-5-fluoro-2,3dihydro-1H-inden-1 -yl)piperazin-1 -yl)-6-chloro-1 -methyl-2-oxo-1,2 -dihydro-1,5-naphthyridine-3-carbonitrile (200); 6-chloro-4-(4-(5-fluoro-7-hydroxy-2,3-dihydro-1H-inden-1-yl)piperazin-1-yl)-1-methyl-2oxo-1,2-dihydro -1,5-naphthyridine-3-carbonitrile (201); 4-(4-benzhydrylpiperazin-1 -yl)-6-ethyl-1-methyl2-oxo-1,2-dihydro-1,5-naphthyridin-3-carbonitrile (202); 4-(4-benzhydrylpiperazin-1 -yl)-1,6-dimethyl2-oxo-1,2-dihydro-1,5-naphthyridin-3-carbonitrile (204); 4-(4-benzhydrylpiperazin-1 -yl)-1-methyl-2oxo-6-vinyl-1,2-dihydro-1,5-naphthyridin-3-carbonitrile (208); 6-chloro-4-(4-(cyclohexyl(phenyl)methyl)piperazin-1 -yl)-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (209); 4-(4-(2-aminobenzyl)piperazin-1-yl)-6-bromo-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3-carbonitrile (211); N-(2((4-(6-bromo-3-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)piperazin-1-yl)methyl)phenyl )methanesulfonamide (212); N-(2-((4-(6-bromo-3-cyano-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)piperazin-1 -yl)methyl) phenyl)benzamide (213); 1 -(2-((4-(6-bromo-3-cyano-1 -methyl-2-oxo-1,2-dihydro-1,5naphthyridin-4-yl)piperazin-1 -yl)methyl)phenyl) -3-phenylurea (214); N-(2-((4-(6-6pom-3-cyano-1 methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)piperazin-1 -yl)methyl)phenyl )acetamide (215); 6-chloro-4(4-(indolin-7-ylmethyl)piperazin-1 -yl)-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3-carbonitrile (216); 8-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2,7-dicarbonitrile (217); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-6-chloro-1-methyl-2-oxo-1,2-dihydro- 268 044500 1,5-naphthyridine-3-carbonitrile (218); 6-chloro-4-(4-((2-hydroxyphenyl)(phenyl)methyl)piperazin-1-yl)-1methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (219 ); 6-chloro-4-(4-(( 1 -ethyl-1 H-indol-4yl)methyl)piperazin-1 -yl)-1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile (220); 6-chloro-1 methyl-4-(4-(naphthalene-1 -ylmethyl)piperazin-1 -yl)-2-oxo-1,2-dihydro-1,5-naphthyridin-3-carbonitrile (221); 6-chloro-4-(4-((4-fluoro-2-hydroxyphenyl)(4-fluorophenyl)methyl)piperazin-1-yl)-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridine -3-carbonitrile (222); 8-(4-benzhydrylpiperazin-1-yl)-5-methyl-7-nitro-6-oxo5,6-dihydro-1,5-naphthyridin-2-carbonitrile (223); 3-bromo-4-(4-(1-(4-fluorophenyl)ethyl)piperazin-1-yl)1,6-dimethyl-1,5-naphthyridin-2(1H)-one (224); 6-bromo-4-(4-(( 1 -ethyl-1H-indol-4-yl)methyl)piperazin-1 -yl)1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile (225); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1 -yl)-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (226); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-1-methyl-3-(2,2,2-trifluoroacetyl)-1,5-naphthyridin-2(1H)-one ( 227); 4-(4(bis(4-fluorophenyl)methyl)piperazin-1 -yl)-1,6-dimethyl-3-(2,2,2-trifluoroacetyl)-1,5-naphthyridin-2(1H)- he (228); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-6-bromo-2-oxo-1-(prop-2-in-1-yl)-1,2-dihydro-1 ,5naphthyridine-3-carbonitrile (229); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-3-bromo-1-methyl-1,5naphthyridin-2(1H)-one (230); 8-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-7-cyano-K,K,5-trimethyl-6oxo-5,6-dihydro-1,5-naphthyridine-2-carboxamide (231); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)1,6-dimethyl-3-(trifluoromethyl)-1,5-naphthyridin-2(1H)-one (232); 4-(4-(bis(4-fluorophenyl)methyl)piperazin1 -yl)-3-chloro-1,6-dimethyl-1,5-naphthyridin-2(1H)-one (233); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1 -yl)3-fluoro-1,6-dimethyl-1,5-naphthyridin-2(1H)-one (234); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-6bromo-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile, TFU (235 ); 3 -bromo-4-(4-(( 1 -ethyl-1Hindol-4-yl)methyl)piperazin-1 -yl)-1 -methyl-1,5-naphthyridin-2(1 H)-one (236) ; 6-bromo-1-methyl-4-(4(naphthalene-1-ylmethyl)piperazin-1-yl)-2-oxo-1,2-dihydro-1,5-naphthyridin-3-carbonitrile (237); 4-(4([1,1'-biphenyl]-2-ylmethyl)piperazin-1-yl)-6-bromo-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3 -carbonitrile, TFU (238); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1 -yl)-3-bromo-1,6-dimethyl-1,5-naphthyridin2(1H)-one (239); methyl 8-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-7-cyano-5-methyl-6-oxo-5,6dihydro-1,5-naphthyridine-2-carboxylate (240) ; 8-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-7-cyano-5methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2-carboxylic acid (241) ; 4-[4-(diphenylmethyl)piperazin1 -yl]-1 -methyl-3-nitro-1,2-dihydro-1,5-naphthyridin-2-one (242); 4-[4-(diphenylmethyl)piperazin-1-yl]-1ethyl-3-nitro-1,2-dihydro-1,5-naphthyridin-2-one (243); 4-[4-(diphenylmethyl)piperazin-1-yl]-1-(2methoxyethyl)-3-nitro-1,2-dihydro-1,5-naphthyridin-2-one (244); 2-{4-[4-(diphenylmethyl)piperazin-1-yl]3-nitro-2-oxo-1,2-dihydro-1,5-naphthyridin-1-yl}acetonitrile (245); ethyl 2-{4-[4-(diphenylmethyl)piperazin-1 -yl]-3-nitro-2-oxo-1,2-dihydro-1,5-naphthyridin-1 -yl} acetate (246); 4-[4-(diphenylmethyl)piperazin-1-yl]-3-nitro-1-propyl-1,2-dihydro-1,5-naphthyridin-2-one (248); 4-{4-[cyclopropyl(4-fluorophenyl)methyl]piperazin-1-yl}-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (250); 4-{4[bis(4-fluoro-2-methoxyphenyl)methyl]piperazin-1-yl} - 1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3 carbonitrile (251 ); 4-[4-(4-methoxybutan-2-yl)piperazin-1 -yl]-1,6-dimethyl-2-oxo-1,2-dihydro-1,5naphthyridin-3-carbonitrile (252); 4-[4-(3,4-dihydro-2H-1 -benzopyran-4-yl)piperazin-1 -yl]-1,6-dimethyl2-oxo-1,2-dihydro-1,5-naphthyridin-3 -carbonitrile (253); 4-{4-[(4-fluorophenyl)(2-methoxypyridin-3yl)methyl]piperazin-1-yl}-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3 -carbonitrile (254); 4-{4[(4-fluorophenyl)(3-methoxypyridin-2-yl)methyl]piperazin-1-yl}-1,6-dimethyl-2-oxo-1,2-dihydro-1,5naphthyridin-3- carbonitrile (255); 4-{4-[(4-fluorophenyl)(pyridin-2-yl)methyl]piperazin-1-yl}-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3 -carbonitrile (256); 4-{4-[(2-bromo-6-hydroxyphenyl)methyl]piperazin-1-yl}-6-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (258); 6-bromo-4-{4-[(2hydroxy-6-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 259); 8-{4-[2-(4-fluorophenyl)propan-2-yl]piperazin-1-yl}-5-methyl-7-nitro-6-oxo-5,6-dihydro1,5-naphthyridin-2- carbonitrile (260); 4-{4-[(4-tert-butyl-2-hydroxyphenyl)methyl]piperazin-1-yl}-6chloro-1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3 - carbonitrile (261); 4-(4-{ [2-hydroxy-5-(trifluoromethoxy)phenyl]methyl}piperazin-1 -yl)-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3 -carbonitrile (262); 8-{4-[(4-bromo-2-hydroxyphenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro-1,5naphthyridine-2,7-dicarbonitrile (263) ; 6-chloro-4-{4-[(2-hydroxy-6-methoxyphenyl)methyl]piperazin-1-yl}1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (264); 8-(4-{[2-hydroxy-4-(trifluoromethoxy)phenyl]methyl}piperazin-1-yl)-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2,7 -dicarbonitrile (265); 6-bromo-4-{4-[(4-chloro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile ( 266); 6-chloro-4-{4-[(2-chloro-6-hydroxyphenyl)methyl]piperazin-1-yl}-1methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 267); 4-{4-[(3-bromo-2-hydroxyphenyl)methyl]piperazin-1-yl}-6-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (268); 8-{4-[(4chloro-2-hydroxyphenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2,7-dicarbonitrile (269) ; 6-bromo-4-(4-{[2-hydroxy-5-(trifluoromethoxy)phenyl]methyl}piperazin-1-yl)-1-methyl-2oxo-1,2-dihydro-1,5-naphthyridin-3 -carbonitrile (270); 6-bromo-4-{4-[(2-bromo-6-hydroxyphenyl)methyl]piperazin-1 -yl} -1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (271); 6-chloro-4-{4-[(2chlorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (272); 6chloro-4-{4-[(2-hydroxy-4-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthy- 269 044500 ridin- 3-carbonitrile (273); 6-chloro-4-{4-[(3-fluoro-2-methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 274); 4-{4-[(5-bromo-2-hydroxyphenyl)methyl]piperazin-1-yl}-6-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (275); 4-{4-[(4-bromo-2hydroxyphenyl)methyl]piperazin-1-yl}-6-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 276); 4-{4-[(5-bromo-2-hydroxyphenyl)methyl]piperazin-1-yl}-1,6-dimethyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile (277) ; 6-bromo-4-{4-[(3-chloro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 278); 6-chloro-4-{4-[(3-fluoro-4-methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (279); 6-chloro-4{4-[(2-fluoro-6-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3carbonitrile (280 ); 6-chloro-4-{4-[(2-fluoro-3-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridine-3-carbonitrile ( 281); 6-bromo-4-{4-[(5-bromo-2-hydroxyphenyl)methyl]piperazin1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 282); 6-chloro-4-{4-[(2-hydroxy-5methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 283); tert-butyl N-(2-{ [4-(6-bromo-3-cyano-1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-4-yl)piperazin-1 yl] methyl}phenyl)carbamate (284); 6-chloro-1-methyl-2-oxo-4-(4-{[2-(trifluoromethoxy)phenyl]methyl}piperazin-1-yl)-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (285); 4-{4-[(3-bromo-2-hydroxyphenyl)methyl]piperazin-1-yl}-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 286); 6-chloro-4-{4[(2-chloro-4-fluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3carbonitrile (287 ); 6-chloro-4-{4-[(4-chloro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridine-3-carbonitrile ( 288); 6-chloro-4-{4-[(3-fluoro-4-methylphenyl)methyl]piperazin-1yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 289); 6-chloro-4-{4-[(3-fluoro-5-methylphenyl)methyl]piperazin-1 -yl} -1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3 - carbonitrile (290); 6-chloro4-{4-[(3,5-difluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (291) ; 6-chloro-4-{4-[(2-hydroxy-6-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridine-3-carbonitrile ( 292); 6-chloro-4-{4-[(3-chloro-2-hydroxyphenyl)methyl]piperazin1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 293); 6-bromo-4-{4-[(2-fluoro-6-hydroxyphenyl)methyl]piperazin-1 -yl} -1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (294); 6bromo-4-{4-[(2-hydroxy-4-methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 295); 4-{4-[(5-chloro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1,6-dimethyl-2-oxo1,2-dihydro-1,5-naphthyridine-3-carbonitrile (296) ; 6-bromo-4-{4-[(2-methoxyphenyl)methyl]piperazin-1yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (297); 6-chloro-4-{4-[(3,5-difluoro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine- 3-carbonitrile (298); 6bromo-4-{4-[(2,3-dihydro-1 H-indol-7-yl)methyl] piperazin-1 -yl} -1 -methyl-2-oxo-1,2-dihydro-1,5 -naphthyridine-3-carbonitrile (299); 6-bromo-4-{4-[(2-chlorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridine-3-carbonitrile (300); 6-chloro-4-{4-[(2-hydroxy-4-methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (301); 6-chloro-4-{4-[(3-chlorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (302) ; 6-chloro4-{4-[(1 H-indol-7-yl)methyl]piperazin-1 -yl}-1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (303); 6-bromo-4-{4-[(3-fluoro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro1,5-naphthyridine-3-carbonitrile ( 304); 6-chloro-4-{4-[(2-hydroxy-5-methoxyphenyl)methyl]piperazin-1-yl}1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (305); 6-chloro-4-{4-[(3-fluoro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (306); 6-chloro-4{4-[(3,5-dichloro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3 carbonitrile (307); 6-chloro-4-{4-[(4-fluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile (308); 6-bromo-4-{4-[(3-tert-butyl-2-hydroxyphenyl)methyl]piperazin-1-yl}1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3 -carbonitrile (309); 6-chloro-4-{4-[(2,4-dichlorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 310); 6-chloro-4-(4-{[2hydroxy-4-(trifluoromethyl)phenyl]methyl}piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine3-carbonitrile (311); 6-chloro-4-{4-[(5-fluoro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridine-3-carbonitrile ( 312); 6-chloro-4-(4-{[3-fluoro-5-(trifluoromethyl)phenyl]methyl}piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile (313); 6-bromo-4-{4-[(2hydroxy-6-methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 314); 6-chloro-4-{4-[1-(4-fluorophenyl)-2-methylpropyl]piperazin-1-yl}-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridine-3-carbonitrile (315-316); 8-{4-[(2-hydroxyphenyl)methyl]piperazin-1-yl}-5methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2,7-dicarbonitrile (317); 6-chloro-4-{4-[(2,5-difluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 318); 6-chloro-4-{4[(3,4-difluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (319 ); 6-bromo-4-{4-[(3,5-difluoro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro1,5-naphthyridin-3- carbonitrile (320); 6-chloro-4-{4-[(3-fluorophenyl)methyl]piperazin-1-yl}-1-methyl-2oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (321); 6-chloro-4-{4-[(1H-indazol-7-yl)methyl]piperazin1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3-carbonitrile ( 322); 6-chloro-4-{4-[(4-chloro-3hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile - 270 044500 (323); 6-chloro-4-{4-[(3-chloro-5-fluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile ( 324); 8-[4-(diphenylmethyl)piperazin-1-yl]-5-methyl-6-oxo-5,6-dihydro-1,5naphthyridine-2,7-dicarbonitrile (325); 6-chloro-4-{4-[(4-chloro-3-fluorophenyl)methyl]piperazin-1-yl}-1methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 326); 6-chloro-4-{4-[(3-methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (327) ; 6-chloro-4-{4-[(3-hydroxy-4-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (328); 4-{4-[(3-fluoro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 329); 6-chloro-4-{4-[(2,3-difluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo1,2-dihydro-1,5-naphthyridine-3-carbonitrile (330) ; 6-chloro-4-{4-[(2-methoxyphenyl)methyl]piperazin-1yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (331); 8-{4-[(4-chloro-3-hydroxyphenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2,7-dicarbonitrile ( 332); 6bromo-4-{4-[(1 H-indazol-7-yl)methyl]piperazin-1 -yl}-1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (333); 4-{4-[(1-ethyl-1H-indol-4-yl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridin-3-carbonitrile ( 334); 6-chloro-4-{4-[(2-fluoro-6-methoxyphenyl)methyl]piperazin-1-yl}-1methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 335); 6-chloro-4-{4-[(2-hydroxynaphthalene-1yl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (336); 4-{4-[(2hydroxyphenyl)methyl]piperazin-1-yl}-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3-carbonitrile (337); 6-chloro-4-{4-[(3-fluoro-5-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile ( 338); 6-chloro-4-{4-[(2-fluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo1,2-dihydro-1,5-naphthyridine-3-carbonitrile (339); 6-chloro-4-{4-[(5-cyano-2-fluorophenyl)methyl]piperazin1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 340); 6-chloro-4-{4-[(4-methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (341) ; 8-{4[1-(4-fluorophenyl)cyclopropyl]piperazin-1-yl}-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine-2carbonitrile (342); 4-(4-benzylpiperazin-1 -yl)-6-chloro-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3carbonitrile (343); 6-bromo-4-{4-[(2-fluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile (344); 6-chloro-4-{4-[(3-chloro-5-hydroxyphenyl)methyl]piperazin-1-yl}-1methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 345); 6-chloro-1-methyl-4-{4-[(2-methylphenyl)methyl]piperazin-1-yl}-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (346) ; 4-{4-[bis(4-fluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (347); 6-chloro-4{4-[(4-fluoro-3-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (348); 4-{4-[(2-hydroxyphenyl)(phenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile (349); 6-chloro-4-{4-[(2,4-difluorophenyl)methyl]piperazin-1-yl}-1-methyl-2oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (350) ; 6-chloro-4-{4-[(3-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (351) ; 4-{4-[(2-hydroxy-3 methoxyphenyl)methyl]piperazin-1-yl}-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (352 ); 6-chloro-4-{4-[(3-hydroxy-4-methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile ( 353); 6-chloro-4-{4-[(3-chloro-4-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3 - carbonitrile (354); 6-chloro-4-(4-{[4-hydroxy-3 -(trifluoromethyl)phenyl]methyl}piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile (355); 6-chloro-4-{4-[(4-hydroxy-3-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (356); 6-chloro-4-{4-[(3-fluoro-4-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 357); 6-chloro-4-[4-(diphenylmethyl)piperazin-1 -yl]-1 methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (358); 4-{4-[(1H-1,3-benzodiazol-7-yl)methyl]piperazin-1 -yl}-6-chloro-1-methyl-2-oxo-1,2-dihydro-1,5- naphthyridine-3-carbonitrile (359); 4-{4[(1H-1,3-benzodiazol-7-yl)methyl]piperazin-1 -yl}-6-bromo-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine3 -carbonitrile (360); 6-chloro-4-{4-[(4-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (362) ; 8-{4-[(3-fluoro-4-hydroxyphenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2,7-dicarbonitrile ( 363); 6-chloro-4-{4-[(2-fluoro-5-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (364); 6-chloro-4{4-[(2-chloro-6-methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3carbonitrile (365 ); 6-bromo-4-{4-[(3,5-dichloro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo1,2-dihydro-1,5-naphthyridin-3- carbonitrile (366); 6-bromo-4-{4-[(5-chloro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (367); 6-bromo-4-{4-[(2-hydroxy3-methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 368); 6-chloro-4-(4-{[2-hydroxy-5-(trifluoromethoxy)phenyl]methyl}piperazin-1-yl)-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridin-3 -carbonitrile (369); 6-chloro-4-{4-[(5-chloro-2-hydroxyphenyl)methyl]piperazin1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 370); 6-chloro-4-{4-[(2-hydroxy-3methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 371); 6-bromo-4-{4-[(2-hydroxy-4-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (372); 6-bromo-4-{4-[(1H-indol-7-yl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridin-3-carbonitrile ( 373); 6-bromo-1-methyl-2-oxo-4-{4-[(2-oxo-2,3-dihydro-1Hindol-7-yl)methyl]piperazin-1 -yl}-1,2-dihydro- 1,5-naphthyridine-3-carbonitrile (374); 6-chloro-4-(4-{[3 - 271 044500 fluoro-4-(trifluoromethyl)phenyl]methyl}piperazin-1-yl)-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (375); 6-chloro-1-methyl-2-oxo-4-{4-[(2-oxo-2,3-dihydro-1H-indol-7-yl)methyl]piperazin-1-yl}1,2-dihydro -1,5-naphthyridine-3-carbonitrile (376); 6-chloro-4-{4-[1-(4-fluorophenyl)-3,3-dimethylbutyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile (377-378); 6-chloro-4-{4-[(4cyano-2-fluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 379); 6-bromo-4-{4-[(3-bromo-2-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile ( 380); 6-chloro-4-(4-{[2-hydroxy-4-(trifluoromethoxy)phenyl]methyl}piperazin1-yl)-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3 -carbonitrile (381); 6-chloro-4-{4-[cyclobutyl(4fluorophenyl)methyl]piperazin-1 -yl}-1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (382383); 4-{4-[(3-tert-butyl-2-hydroxyphenyl)methyl]piperazin-1-yl}-6-chloro-1-methyl-2-oxo-1,2-dihydro1,5-naphthyridin-3- carbonitrile (384); 6-chloro-1-methyl-4-{4-[(3-methylphenyl)methyl]piperazin-1-yl}-2oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (385); 6-chloro-1-methyl-4-{4-[(4-methylphenyl)methyl]piperazin-1-yl}-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (386) ; 6-chloro-4-{4-[1-(4-fluorophenyl)ethyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 387-388); 4-{4[(3-chloro-2-hydroxyphenyl)methyl]piperazin-1-yl}-1,6-dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3carbonitrile (389); 6-bromo-4-{4-[(2-chloro-6-hydroxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2dihydro-1,5-naphthyridine-3-carbonitrile ( 390); 6-chloro-4-{4-[(2-hydroxy-3-methylphenyl)methyl]piperazin1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 391); 8-{4-[(4-fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridin-2 -carbonitrile (392); 4-{4-[bis(4-chlorophenyl)methyl]piperazin-1-yl}-6-bromo-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (393 ); 8-{4-[bis(4-chlorophenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro1,5-naphthyridine-2,7-dicarbonitrile (394); 4-{4-[bis(4-chlorophenyl)methyl]piperazin-1-yl}-6-methoxy-1methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (395); 4-{4-[(4-fluorophenyl)(2-methoxypyridin3-yl)methyl]piperazin-1-yl}-6-methoxy-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile (396); 4-{4-[(4-fluorophenyl)(3-methoxypyridin-2-yl)methyl]piperazin-1-yl}-6-methoxy-1-methyl-2-oxo1,2-dihydro-1,5-naphthyridine -3-carbonitrile (397); 4-{4-[1-(4-fluorophenyl)propyl]piperazin-1-yl}-1,6dimethyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (398); 8-{4-[(S)-(4-chlorophenyl)(phenyl)methyl]piperazin-1-yl}-5-methyl-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridine -2-carbonitrile (399); 8-{4-[(4fluorophenyl)(pyridin-2-yl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2,7-dicarbonitrile (400); 4-{4-[(4-fluoro-2-methoxyphenyl)(pyrimidin-2-yl)methyl]piperazin-1-yl}-6-methoxy-1methyl-2-oxo-1,2-dihydro-1,5 -naphthyridine-3-carbonitrile (401); 4-{4-[bis(4-fluoro-2-methoxyphenyl)methyl]piperazin-1-yl}-6-methoxy-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3 -carbonitrile (402); 4-{4[(4-fluorophenyl)(pyridin-2-yl)methyl]piperazin-1-yl}-6-methoxy-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine- 3-carbonitrile (403); 5-methyl-8-{4-[(naphthalene-1-yl)methyl]piperazin-1-yl}-6-oxo-5,6-dihydro1,5-naphthyridin-2,7-dicarbonitrile (404); 8-{4-[(4-fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridin-2 ,7-dicarbonitrile (405-407); 5-methyl-8-{4[(4-methylphenyl)(phenyl)methyl]piperazin-1-yl}-6-oxo-5,6-dihydro-1,5-naphthyridine-2,7-dicarbonitrile (408) ; 8-{4-[bis(4-fluoro-2-methoxyphenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2,7-dicarbonitrile (409); 8-{4-[(4-fluorophenyl)(3-methoxypyridin-2-yl)methyl]piperazin-1-yl}-5methyl-6-oxo-5,6-dihydro-1,5-naphthyridin-2,7 -dicarbonitrile (410); 8-{4-[(4-fluorophenyl)(phenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2,7-dicarbonitrile (411 ); 4-{4-[(4fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl]piperazin-1-yl}-6-methoxy-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridin-3 -carbonitrile (412); 4-{4-[(4-fluoro-2-methoxyphenyl)(4-fluorophenyl)methyl]piperazin-1-yl}6-methoxy-1-methyl-2-oxo-1,2-dihydro-1,5- naphthyridine-3-carbonitrile (413); 8-{4-[(S)-(4-chlorophenyl)(phenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2,7 -dicarbonitrile (414); 5methyl-8-{4-[(naphthalene-1-yl)methyl]piperazin-1-yl}-7-nitro-6-oxo-5,6-dihydro-1,5-naphthyridin-2-carbonitrile (415) ; 8-{4-[(4-fluorophenyl)(2-methoxy-4-methylphenyl)methyl]piperazin-1-yl}-5-methyl-7-nitro6-oxo-5,6-dihydro-1,5-naphthyridine -2-carbonitrile (416); 6-chloro-4-{4-[(4-chlorophenyl)methyl]piperazin-1yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (417); 8-{4-[(2-hydroxyphenyl)(phenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6-dihydro-1,5-naphthyridine-2,7-dicarbonitrile (418 ); 6chloro-4-{4-[(2-chloro-6-methylphenyl)methyl]piperazin-1 -yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3 carbonitrile (419 ); 6-chloro-4-{4-[(2-chloro-6-fluorophenyl)methyl] piperazin-1 -yl} -1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3 - carbonitrile (420); 6-chloro-4-{4-[1 -(4-fluorophenyl)cyclopropyl]piperazin-1-yl}-1 methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (421 ); 6-chloro-4-{4-[(2,6-difluorophenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 422); 6-chloro-4-{4-[(2fluoro-4-methylphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile ( 423); 6-chloro-4-{4-[(4-cyano-2-methoxyphenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5naphthyridine-3-carbonitrile ( 424); 6-chloro-4-{4-[2-(4-fluorophenyl)propan-2-yl]piperazin-1-yl}-1-methyl-2oxo-1,2-dihydro-1,5-naphthyridin-3- carbonitrile (425); 4-(4-(bis(4-fluorophenyl)methyl)piperazin-1-yl)-1methyl-1,5-naphthyridin-2(1H)-one (426); 4-{4-[bis(4-fluorophenyl)methyl]piperazin-1 -yl}-1,6-dimethyl-1,2dihydro-1,5-naphthyridin-2-one (427); 6-bromo-4-{4-[(S)-(4-chlorophenyl)(phenyl)methyl]piperazin-1-yl}-1methyl-2-oxo-1,2-dihydro-1,5-naphthyridin-3 -carbonitrile (428); 6-bromo-4-{4-[(4-chlorophenyl)(phenyl)me- 272 044500 til]piperazin-1 -yl} -1 -methyl-2-oxo-1,2-dihydro-1,5-naphthyridine -3-carbonitrile (429); 6-bromo-4-{4-[(4fluorophenyl)(phenyl)methyl]piperazin-1-yl}-1-methyl-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (430 ); 6-bromo-1-methyl-4-{4-[(4-methylphenyl)(phenyl)methyl]piperazin-1 -yl}-2-oxo-1,2-dihydro-1,5-naphthyridine-3-carbonitrile (431) or 8-{4-[bis(4-fluorophenyl)methyl]piperazin-1-yl}-5-methyl-6-oxo-5,6dihydro-1,5-naphthyridine-2-carbonitrile (432). 7. A pharmaceutical composition containing a compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. 8. Use of a compound according to any one of claims 1 to 6 for the treatment of cancer or viral infections. 9. Use according to claim 8, characterized in that said cancer is selected from colon cancer, pancreatic cancer, breast cancer, prostate cancer, lung cancer, ovarian cancer, cervical cancer, kidney cancer, head and neck cancer, lymphoma, leukemia and melanoma. 10. Use of a compound according to any one of claims 1 to 6 for inhibiting the activity of at least one of diacylglycerol kinases selected from diacylglycerol kinase alpha (DGKa) and diacylglycerol kinase zeta (ϋΟΚζ).