WO2017088755A1

WO2017088755A1 - Aminopyrimidine heterocyclic compound with adenosine receptor antagonistic activity

Info

Publication number: WO2017088755A1
Application number: PCT/CN2016/106906
Authority: WO
Inventors: Zhang XIAOHU
Original assignee: Suzhou Yunxuan Yiyao Keji Youxian Gongsi
Priority date: 2015-11-23
Filing date: 2016-11-23
Publication date: 2017-06-01
Also published as: CN106749190A; CN106749190B

Abstract

Disclosed hereinis an aminopyrimidine heterocyclic compound with adenosine receptor antagonistic activity, comprising a compound of the general formula (I), or a pharmaceutically acceptable salt thereof. The aminopyrimidine heterocyclic compound with adenosine receptor antagonistic activitydisclosed herein can be used as an effective adenosine receptor antagonist, and can be used for the treatment or prevention of disorders caused by abnormal level of adenosine.

Description

AMINOPYRIMIDINE HETEROCYCLIC COMPOUND WITH ADENOSINE RECEPTOR ANTAGONISTIC ACTIVITY

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of Chinese Patent Application 201510816690.2, filed on November 23, 2015, which is hereby incorporated by reference.

FIELD OF THE INVENTION

Disclosed herein relates to the field of pharmaceutical technology, and particularly relates toan aminopyrimidine heterocyclic compound with adenosine receptor antagonistic activity.

BACKGROUND OF THE INVENTION

Adenosine is a ubiquitous modulator of a number of physiological activities, particularly within the cardiovascular and nervous systems, and regulates a wide range of physiological functionsby interacting with specific cell surface receptors. Known adenosine receptors (ARs) are classified as receptors A1, A2A, A2Band A3, and belong to the G protein-coupled receptor family. Among these, the A1and A3receptors down-regulate cellular cAMP levels through their coupling to G proteins to inhibit adenylate cyclase. In contrast, the A2Aand A2Breceptors up-regulate cellular cAMP levels through their coupling to G proteins to activate adenylate cyclase.

Under normal physiological conditions, A1 ARs are quiescent. However, A1 ARs are upregulated in conditions of stress, such as ischaemia, and in conditions of inflammation. A1 ARs are upregulated and activated in airway epithelium and bronchial smooth muscle in human asthmatics. Activation of A1 ARs induces the release of mediators and cytokines that lead to airway hyperreactivity, inflammation and airway remodeling, produces bronchoconstriction in human asthmatic bronchial tissue. Thus, the A1 AR antagonists can play potential therapeutic role in inflammatory diseases and asthma. In addition, A1 antagonists have also been shown to have therapeutic potential in diseases such as hypertension, congestive heart failure.

The A2B adenosine receptor subtype (see Feoktistov, I., Biaggioni, I., Pharmacol. Rev. 1997, 49, 381-402) has been identified in a variety of human and murine tissues and is involved in the regulation of various physiological activities. For example, the binding of adenosine to A2B receptorsstimulates angiogenesis by promoting the growth ofendothelialcells. However, the hyperproliferation ofendothelialcellspromotesdiabeticretinopathy, and an undesirable increase in blood vessels occurs in neoplasia. Accordingly, antagonistsofadenosine A2Breceptors will alleviate or prevent hypervasculation, thus preventingretinopathyand inhibiting tumor formation. In the gastrointestinal tract and metabolic systems, A2B adenosine receptor subtypes appear to be associated with regulation of hepatic glucose production, modulation of intestinal motility and intestinal secretion. Therefore, A2B antagonists may be helpful for the treatment of type II diabetes and obesity. In addition, type I hypersensitivity disorders, suchasasthma, hayfever, andatopiceczema, are stimulated by binding to A2B-receptorsof mast cells. Therefore, blocking these adenosinereceptorswould provide a therapeutic benefit againstsuchdisorders.

It is found that activation of A3 receptors can trigger mast cell degranulation, and promote the release of vasoconstrictor substances, resulting in desensitization and hypotension responses, and is associated with the deterioration of motion and the desensitization of receptor. Thus, A3 receptor antagonists have been recommended for development as anti-asthma drugs (Fishman and Bar-Yehuda, 2003； Nadeem and Mustafa, 2006) . A3 receptor antagonists have also been shown to play therapeutic role in various diseases including cardio-protection (Vasc. Pharmacol., 2005, 42, 271； J. Pharm. Exp. Ther., 2006, 319, 1200) and cancer (WO200010391) .

Adenosine A2A receptors mainly distribute in striatum, modulate the release of GABA in the striatum, which possibly regulates the function of medium spiny neurons. Studies on genetically modified mice and pharmacological analysis suggest that A2Areceptor is a promising therapeutic target for the treatment of central nervous system (CNS) disorders and diseases such as Parkinson's disease, Huntington's disease, attention deficit hyperactivity disorders (ADHD) , stroke, and Alzheimer's disease (Fredholm et al., Annu. Rev. Pharmacol. Toxicol. 2005 45: 385-412； Higgins et al., Behav. Brain Res. 2007 185: 32-42； Dall'Igna et al., Exp Neurol. 2007, 203 (1) : 241-5； Arendash et al., Neuroscience 2006, 142 (4) : 941-52) but also for various psychoses of organic origin (Weiss et al., Neurology. 200, 61: S88-93) . Thus, A2Areceptor antagonists may be a useful treatment for neurodegenerative movement disorders such as Parkinson and Huntington's disease (Tuite P, et al., J. Expert Opin Investig Drugs. 2003, 12: 1335-52； Popoli P. et al., J. Neurosci. 2002, 22: 1967-75) , restless leg syndrome (Happe S, et al., Neuropsychobiology. 2003, 48: 82-6) , and dyskinesias such as those caused by prolonged use of neuroleptic and dopaminergic drugs (Jenner P. J. Neurol. 2000, 247 Suppl2: II43-50) . In addition, A2A antagonists may have therapeutic potential that be used as neuroprotective agents (Stone TW, Drug Dev. Res. 2001, 52: 323-330) , and for the treatment of sleep disorders (Dunwiddie TV., Ann. Rev. Neurosci. 2001, 24: 31-55) .

The immune system is not only responsible for defending its host against microbial invasion, but also can remove the changed host component from organism, where anti-tumor immune mechanism exists. When the immune surveillance function is weakened due to immune system per se or tumor cells, favorable conditions are provided for the development and progression of tumors. It is demonstrated that the hypoxia in tumor tissue would induce the release of higher concentrations of adenosine, and activate T-cell surface A2A adenosine receptor-mediated A2A receptor binding dependent signaling pathway, thereby reduce the release of IFN-γ and overexpress PD-1, CTLA-4, COX2, IL-10 and TGF-β, ultimately lettingtumor cells escape from the attack of the immune system and achieving immune escape (Sitkovsky M. V. et. al., Cancer Immunol. Res., 2014, 2 (7) : 598-605) . Therefore, if the A2A adenosine receptor antagonists are used for inhibiting A2A adenosine receptors on lymphocyte surface, the anti-tumor immunity in tumor microenvironment would be enhanced, thereby controlling and killing tumor cells. The A2A adenosine receptor antagonists in combination with other cancer immunotherapy, such as immune checkpoint monoclonal antibody, can enhance the killing effect on tumor cells (Comput. Struct. Biotechnol. J., 2015, 13: 265-2672) .

As can be seen from above, there is a need to provide effective adenosine receptor antagonists in order to improve disorders caused by the abnormal level of adenosine mentioned above.

SUMMARY OF THE INVENTION

In view of the drawbacks of the prior art discussed above, disclosed herein is to provide an aminopyrimidine heterocyclic compound, which can be used as an effective adenosine receptor antagonist, and can be used for the treatment or prevention of disorders caused by abnormal level of adenosine.

The object of the present invention will be achieved by the following technical solutions: An aminopyrimidine heterocyclic compound with adenosine receptor antagonistic activity, comprising the compound of the general formula (I) ,

wherein:

A₁, A₂, A₃, and A₄ are each independently selected from the group consisting of N and CR₆；

R₁ is aryl or 5-6 membered heteroaryl comprising 1-3 heteroatoms independently selected from N, O and S, wherein said aryl or 5-6 membered heteroaryl is unsubstituted or substituted by 1-3 R₇ groups；

R₂ is independently selected from the group consisting of cyano and halo；

R₃ is independently selected from the group consisting of halo, cyano, C_1-6 alkyl, C_3-6 cycloalkyl, C_2-6 alkenyl, C_2-6 alkynyl, C_2-6 ureido, C_2-6 oxoureido, C_1-6 alkoxy, C_1-6 acyl, NR₄R₅ and CONR₄R₅；

R₄ andR₅ are each independently selected from the group consisting of hydrogen, C_1-6 alkyl, C_3-6 cycloalkyl, and C_1-6acyl, wherein said alkyl, cycloalkyl, and acyl may be substituted by 1-3 R₈ groups； or R₄ and R₅, together with the nitrogen atom to which they are attached, may form a saturated heterocycle comprising 1-3 heteroatoms independently selected from the group consisting of N, O and S, and said saturated heterocycle may be substituted by 1-3 R₉ groups, and two R₉ substituents may, together with the atom to which they are attached, also form a ring；

R₆ and R₇ are each independently selected from the group consisting of hydrogen, deuterium, halo, hydroxy, cyano, C_3-6 cycloalkyl, C_1-6 alkoxy, and C_1-6 alkyl unsubstituted or substituted by halo or deuterium；

R₈ and R₉ are each independently selected from the group consisting of hydrogen, deuterium, halo, hydroxy, cyano, morpholino, piperazin-1-yl, C_3-6 cycloalkyl, C_1-6 alkoxy, and C_1-6 alkyl unsubstituted or substituted by halo or deuterium；

or a pharmaceutically acceptable salt thereof.

In the aminopyrimidine heterocyclic compound with adenosine receptor antagonistic activity described above, preferably, it is the compound of the general formula (II) ,

wherein:

or a pharmaceutically acceptable salt thereof.

In the aminopyrimidine heterocyclic compound with adenosine receptor antagonistic activity described above, preferably, R₁ includes:

In the aminopyrimidine heterocyclic compound with adenosine receptor antagonistic activity described above, preferably, it is the compound of the general formula (IIA) ,

wherein:

R₁ is independently selected from

A₁, A₂, A₃, and A₄ are each independently selected from the group consisting of N and CR₆； wherein A₁, A₂, A₃, and A₄ are each CR₆； alternatively, one of A₁, A₂, A₃, and A₄ is N, the others are each CR₆；

R₄ and R₅ are each independently selected from the group consisting of hydrogen, C_1-6 alkyl, C_3-6 cycloalkyl, and C_1-6acyl, wherein said alkyl, cycloalkyl, and acyl may be substituted by 1-3 R₈ groups； or R₄ and R₅, together with the nitrogen atom to which they are attached, may form a saturated heterocycle comprising 1-3 heteroatoms independently selected from the group consisting of N, O and S, and said saturated heterocycle may be substituted by 1-3 R₉ groups, and two R₉ substituents may, together with the atom to which they are attached, also form a ring；

R₁₀, R₁₁, and R₁₂ are each independently selected from the group consisting of hydrogen, deuterium, halo, cyano, C_3-6 cycloalkyl, and C_1-6 alkyl unsubstituted or substituted by halo or deuterium； or a pharmaceutically acceptable salt thereof.

In the aminopyrimidine heterocyclic compound with adenosine receptor antagonistic activity described above, preferably, it is the compound of the general formula (IIB) ,

wherein:

R₁ is independently selected from

R₃ is NR₄R₅；

R₄ and R₅ are each independently selected from the group consisting of hydrogen, C_1-6 alkyl, C_3-6 cycloalkyl, and C_1-6 acyl, wherein said alkyl, cycloalkyl, and acyl may be substituted by 1-3 R₈ groups； or R₄ and R₅, together with the nitrogen atom to which they are attached, may form a saturated heterocycle comprising 1-3 heteroatoms independently selected from the group consisting of N, O and S, and said saturated heterocycle may be substituted by 1-3 R₉ groups, and two R₉ substituents may, together with the atom to which they are attached, also form a ring；

R₁₀, R₁₁, and R₁₂ are each independently selected from the group consisting of hydrogen, deuterium, halo, cyano, C_3-6 cycloalkyl, and C_1-6 alkyl unsubstituted or substituted by halo or deuterium；

or a pharmaceutically acceptable salt thereof.

In the aminopyrimidine heterocyclic compound with adenosine receptor antagonistic activity described above, preferably, R₃ includes:

In the aminopyrimidine heterocyclic compound with adenosine receptor antagonistic activity described above, preferably, it includes the following Example shown in Table 1 and possible enantiomers,

or a the pharmaceutically acceptable salt thereof.

Table 1: Compounds of Formula I

The present invention further provides a pharmaceutical composition comprising, an aminopyrimidine heterocyclic compound with adenosine receptor antagonistic activity disclosed herein, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier or diluent.

The present invention further provides a combination comprising an aminopyrimidine heterocyclic compound with adenosine receptor antagonistic activity, the combination is a combination of an aminopyrimidine heterocyclic compound with adenosine receptor antagonistic activitydisclosed herein with one or more of L-DOPA, dopamine agonists, dopamine decarboxylase inhibitors, catechol-O-methyltransferase inhibitors and monoamine oxidase inhibitors, cancer immunotherapy such as cancer vaccines, immune checkpoint inhibitors such as cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) and programmed cell death protein 1 (PD-1) .

The present invention further provides use of an aminopyrimidine heterocyclic compound with adenosine receptor antagonistic activity or a combination comprising aminopyrimidine heterocyclic compound with adenosine receptor antagonistic activity in the manufacture of a medicament for antagonising an adenosine receptor.

The aminopyrimidine heterocyclic compound with adenosine receptor antagonistic activity disclosed herein is useful in the treatment of pathological conditions or diseases susceptible of being improved by antagonism of an adenosine receptor, in particular by antagonism of the A2Aadenosine receptor. The diseases or conditions susceptible of being improved by antagonism of an adenosine receptor include, but not limited to, tumor, Alzheimer's disease, Parkinson's disease, neuroprotection, schizophrenia, anxiety, pain, respiratory deficiency, depression, asthma, allergy, and psychoactive substance abuse.

The present invention has the following prominent advantages:

The aminopyrimidine heterocyclic compound with adenosine receptor antagonistic activitydisclosed hereincan be used as an effective adenosine receptor antagonist, and can be used for the treatment or prevention of disorders caused by abnormal level of adenosine.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts the inhibition curve of compound B10 of the present invention in the binding assay；

FIG. 2 depicts the inhibition curve of compound B16 of the present invention in the binding assay；

FIG. 3 depicts the inhibition histogram of compound Bl of the present invention in the function assay；

FIG. 4 depicts the inhibition histogram of compound Bl5 of the present invention in the function assay.

DETAILED DESCRIPTION OF THE INVENTION

The examples below are intended to be purely exemplary and should not be considered to be limiting in any way. Unless otherwise specified, the experimental methods in the Examples describe below are all conventional methods. Unless otherwise specified, the reagents and materials all can be obtained from commercial sources. All solvents and chemicals employed are of analytical grade or chemical purity. Solvents are all redistilled before use. Anhydrous solvents are all prepared according to standard methods or reference methods. Silicagel (100-200 meshes) for column chromatography and silica gel (GF254) for thin-layer chromatography (TLC) were commercially available fromTsingdao Haiyang ChemicalCo., Ltd. or Yantai Chemical Co., Ltd. ofChina； unless otherwise specified, all were eluted with petroleum ether (60-90℃) /ethyl aceate (v/v) , and visualized by iodine or the solution of molybdphosphoric acid in ethanol. All extraction solvents, unless otherwise specified, are dried over anhydrous Na₂SO₄. ¹H NMR spectra are recorded on Bruck-400nuclear magnetic resonancespectrometer with TMS (tetramethylsilane) as theinternalstandard. LC/MSdataarerecorded by using Agilent1100 High Performance Liquid Chromatography-Ion Trap Mass Spectrometor (LC-MSD Trap) equipped with a diode array detector (DAD) detected at 214 nm and 254 nm, and an ion trap (ESI source) . HPLC column: Agela Durashell C18 (4.6×50 mm, 3.5 μm) ； mobile phase: 0.1％aqueous NH₄HCO₃ solution: acetonitrile (from 5: 95 to 95: 5 within 5 minute) ； flow rate: 1.8 mL/min.

Compound B1: 4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (6- (4-methoxypiperidin-1-yl) pyridin-2-yl) pyrimidine-5-carbonitrile

Step A : 4-amino-6- (6-bromopyridin-2-yl) -2- (methylthio) pyrimidine-5-carbonitrile

A mixture of 6-bromopicolinaldehyde (1.86 g, 10.0 mmol) , S-methyl carbamimidothioate H₂SO₄ salt (1.39 g, 10.0 mmol) ， malononitrile (990 mg, 15.0 mmol) and NaOH (200 mg, 5 mmol) was dissolved with EtOH (50 mL) . The resulted mixture was stirred at 50 ℃ for 16 hrs. The reaction mixture was cooled to rt and filtered. The filter cake was washed with 50 mL of H₂O, dried under lamp to give the title product (2.0 g, yield: 62％) as a white solid. MS: M/e 322 (M+1) ⁺.

Step B : 4-amino-6- (6-bromopyridin-2-yl) -2- (methylsulfonyl) pyrimidine -5-carbonitrile

To a suspension of the product from step A (2.00 g, 6.21 mmol) in a mixed solvent (THF/H₂O, 10: 1, 20 mL) was added oxone (4.80 g, 15.6 mmol) . The mixture was stirred at rt for 4 hs. 200 mL of H₂O was added. A gray solid was precipitated and was filtered. The filter cake was dried under lamp to give the title product (1.83 g, crude) as a white solid.

Step C : 4-amino-6- (6-bromopyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol -1-yl) pyrimidine-5-carbonitrile

To a suspension of the product from step B (1.83 g, 5.17 mmol) in EtOH (50 mL) was added hydrazine hydrate (1.22 g, 85％, 20.7 mmol) at rt and the resulted mixture was refluxed for 2 hrs. The reaction mixture was cooled and was added pentane-2, 4-dione (620 mg, 6.20 mmol) at rt. The mixture was refluxed for 4 hrs. A white solid was precipitated and the suspension was cooled and filtered. The filter cake was washed with EtOH (50 mL) and dried under lamp to give the product (1.55 g, 81％) as a white solid. ¹H NMR (400 MHz, DMSO-d6) δ 8.21 (d, J ＝ 8.0Hz, 1H) , 8.02 (m, 1H) , 7.90 (d, J ＝ 8.0Hz, 1H) , 6.18 (s, 1H) , 5.73 (s, 2H) , 2.65 (s, 3H) , 2.20 (s, 3H) . MS: M/e 370 (M+1) ⁺ .

Step D: 4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (6- (4-methoxypiperidin -1-yl) pyridin-2-yl) pyrimidine-5-carbonitrile

A mixture of the product from step C (322 mg, 1.00 mmol) , 4-methoxypiperidine (300 mg, 2.61 mmol) and DIPEA (320 mg, 2.48 mmol) in NMP (10 mL) was heated at 110 ℃ for 12 hrs. Then the mixture was extracted with EtOAc (10 mL x 2) . The combined organic layers were washed with brine (10 mL x 2) , dried over Na₂SO₄ and concentrated. The residue was purified by column chromatography eluting with MeOH﹕ DCM to give the title product (130 mg, 32％) as a light yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 7.65 –7.63 (m, 2H) , 6.85 (d, J ＝ 7.0 Hz, 1H) , 6.10 (s, 1H) , 6.08 (s, 2H) , 4.20 –4.17 (m, 2H) , 3.53 –3.44 (m, 1H) , 3.40 (s, 3H) , 3.41 –3.65 (m, 2H) , 2.78 (s, 3H) , 2.36 (s, 3H) , 2.03 –2.00 (m, 2H) , 1.67 –1.62 (m, 2H) . MS: M/e 405 (M+1) ⁺.

Compound B2: 4-amino-6- (3-chloro-6- (4-methoxypiperidin-1-yl) pyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile

Step A : 4-amino-6- (3, 6-dichloropyridin-2-yl) -2- (methylthio) pyrimidine -5-carbonitrile

A mixture of 3, 6-dichloropicolinaldehyde (528 mg, 3.0 mmol) , S-methyl carbamimidothioate H₂SO₄ salt (417 mg, 3.0 mmol) ， malononitrile (297 mg, 4.5 mmol) and NaOH (60 mg, 1.5 mmol) was dissolved with EtOH (15 mL) . The resulted mixture was stirred at 50 ℃ for 16 hrs. The reaction mixture was cooled to rt and filtered. The filter cake was washed with 20 mL of H₂O, dried under lamp to give the title product (600 mg， 66％) as a white solid. MS: M/e 312 (M+1) ⁺.

Step B : 4-amino-6- (3, 6-dichl^oropyridin-2-yl) -2- (methylsulfonyl) pyrimidine-5-carbonitrile

To a suspension of the product from step A (600 mg, 1.92 mmol) in a mixed solvent (THF/H₂O, 10: 1, 7 mL) was added oxone ⁽1.45 g, 4.84 mmol) . The mixture ^was stirred at rt for 4 hs. 70 mL of H₂O was added. A gray solid was precipi_tated and was filtered. The filte_r cake was dried under lamp to give the title product (650 mg, crude) as a white solid.

Step C : 4-amino-6- (3, 6-dichloropyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile

To a suspension of the product from step B (650 mg, 1.90 mmol) in EtOH (18 mL) was added hydrazine hydrate (440 mg, 85％, 7.45 mmol) at rt and the resulted mixture was refluxed for 2 hrs. The reaction mixture was cooled and was added pentane-2, 4-dione (223 mg, 2.23 mmol) at rt. The mixture was refluxed for 4 hrs. A white solid was precipitated and the suspension was cooled and filtered. The filter cake was washed with EtOH (50 mL) and dried under lamp to give the product (275 mg, 40％) as a white solid. MS: M/e 360 (M+1) ⁺ .

Step D: 4-amino-6- (3-chloro-6- (4-methoxypiperidin-1-yl) pyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile

A mixture of the product from step C (100 mg, 0.28 mmol) , 4-methoxypiperidine (84 mg, 0.73 mmol) and DIPEA (89 mg, 0.69 mmol) in NMP (3 mL) was heated at 110 ℃ for 12 hrs. Then the mixture was extracted with EtOAc (10 mL x 2) . The combined organic layers were washed with brine (10 mL x 2) , dried over Na₂SO₄ and concentrated. The residue was purified by column chromatography eluting with MeOH﹕ DCM to give the title product (20 mg, 16％) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 7.55 (d, J ＝ 8.8 Hz, 1H) , 6.74 (d, J ＝ 9.2 Hz, 1H) , 6.03 (s, 1H) , 5.95 (br s, 2H) , 4.05–3.93 (m, 2H) , 3.52–3.42 (m, 1H) , 3.38 (s, 3H) , 3.39 –3.30 (m, 2H) , 2.68 (s, 3H) , 2.34 (s, 3H) , 2.02 –1.93 (m, 2H) , 1.71–1.60 (m, 2H) . MS: M/e 439 (M+1) ⁺.

Compound B3: 4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (6- (4-hydroxypiperidin-1-yl) pyridin-2-yl) pyrimidine-5-carbonitrile

A mixture of 4-amino-6- (6-bromopyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile (56 mg, 0.15 mmol) , piperidin-4-ol (45 mg, 0.45 mmol) and DIPEA (40 mg, 0.31mmol) in NMP (1 mL) was heated at 110 ℃ for 12 hrs. Then the mixture was extracted with EtOAc (10 mL x 2) . The combined organic layers were washed with brine (10 mL x 2) , dried over Na₂SO₄ and concentrated. The residue was purified by column chromatography eluting with MeOH﹕ DCM to give the title product (25 mg, 43％) as a yellow solid. ¹H NMR (400 MHz, DMSO-d6) δ 8.41 (br s, 1H) , 7.74 –7.70 (m, 1H) , 7.50 (d, J ＝ 7.2 Hz, 1H) , 7.06 (d, J ＝ 8.6 Hz, 1H) , 6.17 (s, 1H) , 4.76 (d, J ＝ 4.0 Hz, 1H) , 4.20 (d, J ＝ 13.6 Hz, 2H) , 3.74 –3.71 (m, 1H) , 3.21 (t, J ＝11.8 Hz, 2H) , 2.64 (s, 3H) , 2.19 (s, 3H) , 1.86 –1.72 (m, 2H) , 1.41 –1.34 (m, 2H) . MS: M/e 391 (M+1) ⁺.

Compound B4: 4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (6- (piperidin -1-yl) pyridin-2-yl) pyrimidine-5-carbonitrile

A mixture of 4-amino-6- (6-bromopyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile (56 mg, 0.15 mmol) , piperidine (38 mg, 0.45 mmol) and DIPEA (40 mg, 0.31mmol) in NMP (1 mL) was heated at 110 ℃ for 12 hrs. Then the mixture was extracted with EtOAc (10 mL x 2) . The combined organic layers were washed with brine (10 mL x 2) , dried over Na₂SO₄ and concentrated. The residue was purified by column chromatography eluting with MeOH﹕ DCM to give the title product (17 mg, 30％) as a yellow solid. ¹H NMR (400 MHz, DMSO-d6) δ 8.35 (br s, 1H) , 7.70 (d, J ＝ 11.2 Hz, 1H) , 7.48 (d, J ＝ 4.0 Hz, 1H) , 7.02 (t, J ＝ 8.8 Hz, 1H) , 6.17 (s, 1H) , 3.73 –3.68 (m, 4H) , 2.64 (s, 3H) , 2.19 (s, 3H) , 1.67 –1.59 (m, 2H) , 1.59 –1.48 (m, 4H) . MS: M/e 375 (M+1) ⁺.

Compound B5: (S) -4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (6- (3-methoxy pyrrolidin-1-yl) pyridin-2-yl) pyrimidine-5-carbonitrile

Step A: tert-butyl (S) -3-hydroxypyrrolidine-1-carboxylate

To a solution of (S) -pyrrolidin-3-ol HCl salt (1.23 g, 10.0 mmol) and Et₃N (2.20 g, 20.0 mmol) in MeOH (20 mL) was added (Boc) ₂O (2.20 g, 10.1 mmol) at ice bath. The mixture was stirred at rt overnight. The solvent was removed and the residue was extracted with DCM. The organic layers were washed with brine (20 mL x 2) , dried over Na₂SO₄ and concentrated to give the title product (1.90 g) as a colorless oil which was used directly in the next step.

Step B: tert-butyl (S) -3-methoxypyrrolidine-1-carboxylate

To a solution of the product from step A (1.90 g, 10.0 mmol) and MeI (1.70 g, 12.0 mmol) in DMF (20 mL) was added NaH (360 mg, 80％, 12.0 mmol) in portion at rt. The mixture was stirred at rt for 4h. Water was added and was extracted with EA. The organic layers were washed with brine (20 mL x 2) , dried over Na₂SO₄ and concentrated. The residue was purified by column chromatography eluting with PE﹕ EA (5 : 1) to give the title product (1.69 g, 84％) as a brown solid.

Step C: (S) -3-methoxypyrrolidine

To a solution of the product from step B (1.69 g, 8.41 mmol) in EA (5 mL) was added HCl/EA (10 mL) . The mixture was stirred at rt for 6 h. The solvent was removed to give the title product (1.24 g) as a brown oil which was used directly in the next step.

Step D: (S) -4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (6- (3-methoxy pyrrolidin-1-yl) pyridin-2-yl) pyrimidine-5-carbonitrile

A mixture of 4-amino-6- (6-bromopyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile (56 mg, 0.15 mmol) , the product from step 3 (45 mg, 0.33 mmol) and DIPEA (65 mg, 0.50 mmol) in NMP (1 mL) was heated at 110 ℃ for 12 hrs. Then the mixture was extracted with EtOAc (10 mL x 2) . The combined organic layers were washed with brine (10 mL x 2) , dried over Na₂SO₄ and concentrated. The residue was purified by column chromatography eluting with MeOH﹕ DCM to give the title product (24 mg, 41％) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 7.66 –7.54 (m, 2H) , 6.55 (d, J ＝ 7.8 Hz, 1H) , 6.08 (s, 3H) , 4.15–4.14 (m, 1H) , 3.91–3.87 (m, 1H) , 3.75–3.73 (m, 1H) , 3.68–3.65 (m, 2H) , 3.40 (s, 3H) , 2.79 (s, 3H) , 2.36 (s, 3H) , 2.24–2.14 (m, 2H) . MS: M/e 391 (M+1) ⁺.

Compound B6: (R) -4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (6- (3-methoxy pyrrolidin-1-yl) pyridin-2-yl) pyrimidine-5-carbonitrile

Step A: tert-butyl (R) -3-hydroxypyrrolidine-1-carboxylate

To a solution of (R) -pyrrolidin-3-ol HCl salt (1.23 g, 10.0 mmol) and Et₃N (2.20 g, 20.0 mmol) in MeOH (20 mL) was added (Boc) ₂O (2.20 g, 10.1 mmol) at ice bath. The mixture was stirred at rt overnight. The solvent was removed and the residue was extracted with DCM. The organic layers were washed with brine (20 mL x 2) , dried over Na₂SO₄ and concentrated to give the title product (1.89 g) as a colorless oil which was used directly in the next step.

Step B: tert-butyl (R) -3-methoxypyrrolidine-1-carboxylate

To a solution of the product from step A (1.89 g, 10.0 mmol) and MeI (1.70 g, 12.0 mmol) in DMF (20 mL) was added NaH (360 mg, 80％, 12.0 mmol) in portion at rt. The mixture was stirred at rt for 4h. Water was added and was extracted with EA. The organic layers were washed with brine (20 mL x 2) , dried over Na₂SO₄ and concentrated. The residue was purified by column chromatography eluting with PE﹕ EA (5 : 1) to give the title product (1.65 g, 82％) as a brown solid.

Step C: (R) -3-methoxypyrrolidine

To a solution of the product from step B (1.65 g, 8.21 mmol) in EA (5 mL) was added HCl/EA (10 mL) . The mixture was stirred at rt for 6 h. The solvent was removed to give the title product (1.14 g) as a brown oil which was used directly in the next step.

Step D: (R) -4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (6- (3-methoxy pyrrolidin-1-yl) pyridin-2-yl) pyrimidine-5-carbonitrile

A mixture of 4-amino-6- (6-bromopyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile (56 mg, 0.15 mmol) , the product from step C (45 mg, 0.33 mmol) and DIPEA (65 mg, 0.50 mmol) in NMP (1 mL) was heated at 110 ℃ for 12 hrs. Then the mixture was extracted with EtOAc (10 mL x 2) . The combined organic layers were washed with brine (10 mL x 2) , dried over Na₂SO₄ and concentrated. The residue was purified by column chromatography eluting with 1％MeOH﹕ DCM to give the title product (17 mg, 29％) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 7.63 –7.56 (m, 2H) , 6.55 (d, J ＝ 8.0 Hz, 1H) , 6.08 (s, 3H) , 4.14 (s, 1H) , 3.90 –3.87 (m, 1H) , 3.76 –3.70 (m, 1H) , 3.70 –3.61 (m, 2H) , 3.40 (s, 3H) , 2.79 (s, 3H) , 2.36 (s, 3H) , 2.24 –2.16 (m, 1H) , 2.18 –2.09 (m, 1H) . MS: M/e 391 (M+1) ⁺.

Compound B7: 4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (6- (3-hydroxy pyrrolidin-1-yl) pyridin-2-yl) pyrimidine-5-carbonitrile

A mixture of 4-amino-6- (6-bromopyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile (56 mg, 0.15 mmol) , pyrrolidin-3-ol (39 mg, 0.45 mmol) and DIPEA (40 mg, 0.31mmol) in NMP (1 mL) was heated at 110 ℃ for 12 hrs. Then the mixture was extracted with EtOAc (10 mL x 2) . The combined organic layers were washed with brine (10 mL x 2) , dried over Na₂SO₄ and concentrated. The residue was purified by column chromatography eluting with MeOH﹕ DCM to give the title product (20 mg, 35％) as a yellow solid. ¹H NMR (400 MHz, DMSO-d6) δ 8.42 (br s, 1H) , 7.72 –7.68 (m, 1H) , 7.45 (d, J ＝ 7.4 Hz, 1H) , 6.65 (d, J ＝ 8.4 Hz, 1H) , 6.16 (s, 1H) , 4.99 (d, J ＝ 3.0 Hz, 1H) , 4.42 (s, 1H) , 3.62 –3.59 (m, 2H) , 3.48 –3.45 (m, 1H) , 2.65 (s, 3H) , 2.19 (s, 3H) , 2.05 -2.03 (m, 1H) , 1.91 –1.89 (m, 1H) . MS: M/e 377 (M+1) ⁺.

Compound B8:

(S) -4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (6- (3-hydroxypyrrolidin-1-yl) pyridin-2-yl) pyrimid ine-5-carbonitrile

A mixture of 4-amino-6- (6-bromopyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile (56 mg, 0.15 mmol) , (S) -pyrrolidin-3-ol HCl salt (56 mg, 0.45 mmol) and DIPEA (65 mg, 0.50 mmol) in NMP (1 mL) was heated at 110 ℃ for 12 hrs. Then the mixture was extracted with EtOAc (10 mL x 2) . The combined organic layers were washed with brine (10 mL x 2) , dried over Na₂SO₄ and concentrated. The residue was purified by column chromatography eluting with MeOH﹕ DCM to give the title product (12 mg, 21％) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 7.64 –7.55 (m, 2H) , 6.55 (d, J ＝ 8.0 Hz, 1H) , 6.09 (s, 3H) , 4.67 (s, 1H) , 3.89 –3.75 (m, 2H) , 3.72 –3.69 (m, 2H) , 2.78 (s, 3H) , 2.37 (s, 3H) , 2.26 –2.09 (m, 2H) . MS: M/e 377 (M+1) ⁺.

Compound B9:

(R) -4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (6- (3-hydroxypyrrolidin-1-yl) pyridin-2-yl) pyrimidine-5-carbonitrile

A mixture of 4-amino-6- (6-bromopyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile (56 mg, 0.15 mmol) , (R) -pyrrolidin-3-ol HCl salt (56 mg, 0.45 mmol) and DIPEA (65 mg, 0.50 mmol) in NMP (1 mL) was heated at 110 ℃ for 12 hrs. Then the mixture was extracted with EtOAc (10 mL x 2) . The combined organic layers were washed with brine (10 mL x 2) , dried over Na₂SO₄ and concentrated. The residue was purified by column chromatography eluting with MeOH﹕ DCM to give the title product (17 mg, 30％) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 7.67 –7.55 (m, 2H) , 6.55 (d, J ＝ 8.0 Hz, 1H) , 6.09 (s, 3H) , 4.67 (s, 1H) , 3.88 –3.76 (m, 2H) , 3.72 –3.69 (m, 2H) , 2.79 (s, 3H) , 2.37 (s, 3H) , 2.26 –2.10 (m, 2H) . MS: M/e 377 (M+1) ⁺.

Compound

B10: 4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (6- (pyrrolidin-1-yl) pyridin-2-yl) pyrimidine-5-carbonitrile

A mixture of 4-amino-6- (6-bromopyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile (56 mg, 0.15 mmol) , pyrrolidine (56 mg, 0.45 mmol) and DIPEA (40 mg, 0.31mmol) in NMP (1 mL) was heated at 110 ℃ for 12 hrs. Then the mixture was extracted with EtOAc (10 mL x 2) . The combined organic layers were washed with brine (10 mL x 2) , dried over Na₂SO₄ and concentrated. The residue was purified by column chromatography eluting with MeOH﹕ DCM to give the title product (10 mg, 18％) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 7.61 –7.59 (m, 1H) , 7.54 (d, J ＝ 7.6 Hz, 1H) , 6.53 (d, J ＝ 8.0 Hz, 1H) , 6.09 (s, 1H) , 6.07 (s, 2H) , 3.62 (s, 4H) , 2.78 (s, 3H) , 2.36 (s, 3H) , 2.05 (s, 4H) . MS: M/e 361 (M+1) ⁺.

Compound

B11: 4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (6- (3-hydroxyazetidin-1-yl) pyridin-2-yl) pyrimidine-5-carbonitrile

A mixture of 4-amino-6- (6-bromopyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile (56 mg, 0.15 mmol) , azetidin-3-olHCl salt (49 mg, 0.45 mmol) and DIPEA (65 mg, 0.50 mmol) in NMP (1 mL) was heated at 110 ℃ for 12 hrs. Then the mixture was extracted with EtOAc (10 mL x 2) . The combined organic layers were washed with brine (10 mL x 2) , dried over Na₂SO₄ and concentrated. The residue was purified by column chromatography eluting with MeOH﹕ DCM to give the title product (10 mg, 18％) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 7.67 –7.55 (m, 2H) , 6.55 (d, J ＝ 8.0 Hz, 1H) , 6.09 (s, 3H) , 4.67 (s, 1H) , 3.88 –3.76 (m, 2H) , 3.72 –3.69 (m, 2H) , 2.79 (s, 3H) , 2.37 (s, 3H) , 2.26 –2.10 (m, 2H) . MS: M/e 363 (M+1) ⁺.

Compound

B12: 4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (6-morpholinopyridin-2-yl) pyrimidine-5-carbonitrile

A mixture of 4-amino-6- (6-bromopyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile (56 mg, 0.15 mmol) , morpholine (40 mg, 0.45 mmol) and DIPEA (40 mg, 0.31mmol) in NMP (1 mL) was heated at 110 ℃ for 12 hrs. Then the mixture was extracted with EtOAc (10 mL x 2) . The combined organic layers were washed with brine (10 mL x 2) , dried over Na₂SO₄ and concentrated. The residue was purified by column chromatography eluting with 1％MeOH﹕ DCM to give the title product (20 mg, 35％) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 7.69 (d, J ＝ 4.2 Hz, 2H) , 6.88 –6.79 (m, 1H) , 6.09 (s, 3H) , 3.92 –3.84 (m, 4H) , 3.76 –3.69 (m, 4H) , 2.79 (s, 3H) , 2.37 (s, 3H) . MS: M/e 377 (M+1) ⁺.

Compound

B13: 4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (6- ( (2-methoxyethyl) amino) pyridin-2-yl) pyrimidine-5-carbonitrile

Step

A: 4-amino-6- (6- ( (2-methoxyethyl) amino) pyridin-2-yl) -2- (methylthio) pyrimidine-5-carbonitrile

A mixture of 4-amino-6- (6-bromopyridin-2-yl) -2- (methylthio) pyrimidine-5-carbonitrile (322 mg, 1.00 mmol) , 2-methoxyethan-1-amine (225 mg, 3.00 mmol) and DIPEA (155 mg, 1.20 mmol) in NMP (3 mL) was heated at 120 ℃ for 12 hrs in the sealed tube. Then the mixture was extracted with EtOAc (10 mL x 2) . The combined organic layers were washed with brine (10 mL x 2) , dried over Na₂SO₄ and concentrated. The residue was purified by column chromatography eluting with PE﹕ EA (4 : 1) to give the title product (90 mg, 28％) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 7.67 (d, J ＝ 7.2 Hz, 1H) , 7.54 –7.50 (m, 1H) , 6.58 (d, J ＝ 8.0 Hz, 1H) , 5.71 (s, 2H) , 4.95 (s, 1H) , 3.87 –3.74 (m, 2H) , 3.70 –3.60 (m, 2H) , 3.40 (s, 3H) , 2.60 (s, 3H) .

Step B :

4-amino-6- (6- ( (2-methoxyethyl) amino) pyridin-2-yl) -2- (methylsulfonyl) pyrimidine-5-carbonitrile

To a suspension of the product from step A (90 mg, 0.28 mmol) in a mixed solvent (THF/H₂O, 10: 1, 5 mL) was added oxone (318 mg, 0.71 mmol) . The mixture was stirred at rt for 4 hs. 5 mL of H₂O was added. The mixture was extracted with EtOAc (10 mL x 2) . The combined organic layers were washed with brine (10 mL x 2) , dried over Na₂SO₄ and concentrated to give the title product (60 mg, crude) as a white solidwhich was used directly in the next step.

Step C :

4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (6- ( (2-methoxyethyl) amino) pyridin-2-yl) pyrimidine-5-carbonitrile

To a suspension of the product from step B (60 mg, 0.17 mmol) in EtOH (5 mL) was added hydrazine hydrate (50 mg, 85％, 0.85 mmol) at rt and the resulted mixture was refluxed for 2 hrs. The reaction mixture was cooled and was added pentane-2, 4-dione (85 mg, 0.85 mmol) at rt. The mixture was refluxed for 4 hrs. The mixture was concentrated and the residue was purified by column chromatography eluting with 1％MeOH﹕ DCMto give the title product (16 mg, 26％) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 7.61 (d, J ＝ 7.2 Hz, 1H) , 7.55 –7.52 (m, 1H) , 6.60 (d, J ＝ 8.2 Hz, 1H) , 6.08 (s, 3H) , 5.00 –4.97 (m, 1H) , 3.85 –3.81 (m, 2H) , 3.65 (t, J ＝ 5.0 Hz, 2H) , 3.40 (s, 3H) , 2.78 (s, 3H) , 2.35 (s, 3H) . MS: M/e 365 (M+1) ⁺ .

Compound B14:

4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (6- ( (2-hydroxyethyl) (methyl) amino) pyridin-2-yl) pyrimidine-5-carbonitrile

A mixture of 4-amino-6- (6-bromopyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile (56 mg, 0.15 mmol) , 2- (methylamino) ethan-1-ol (34 mg, 0.45 mmol) and DIPEA (40 mg, 0.31mmol) in NMP (1 mL) was heated at 110 ℃ for 12 hrs. Then the mixture was extracted with EtOAc (10 mL x 2) . The combined organic layers were washed with brine (10 mL x 2) , dried over Na₂SO₄ and concentrated. The residue was purified by column chromatography eluting with 1％MeOH﹕ DCM to give the title product (25 mg, 46％) as a yellow solid. ¹H NMR (400 MHz, DMSO-d6) δ 8.43 (s, 1H) , 7.70 (d, J ＝ 4.2 Hz, 1H) , 7.54 (s, 1H) , 7.46 (d, J ＝ 6.8 Hz, 1H) , 6.85 (s, 1H) , 6.16 (s, 1H) , 4.62 (s, 1H) , 3.70 (d, J ＝ 4.0 Hz, 2H) , 3.62 (d, J ＝ 4.0 Hz, 2H) , 3.18 (s, 3H) , 2.65 (s, 3H) , 2.19 (s, 3H) . MS: M/e 365 (M+1) ⁺.

Compound B15:

4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (6- ( (2-methoxyethyl) (methyl) amino) pyridin-2-yl) pyrimidine-5-carbonitrile

A mixture of 4-amino-6- (6-bromopyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile (56 mg, 0.15 mmol) , 2-methoxy-N-methylethan-1-amine (34 mg, 0.45 mmol) and DIPEA (40 mg, 0.31mmol) in NMP (1 mL) was heated at 110 ℃ for 12 hrs. Then the mixture was extracted with EtOAc (10 mL x 2) . The combined organic layers were washed with brine (10 mL x 2) , dried over Na₂SO₄ and concentrated. The residue was purified by column chromatography eluting with 1％MeOH﹕ DCM to give the title product (16 mg, 28％) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 7.65 –7.57 (m, 2H) , 6.71 (d, J ＝ 8.8 Hz, 1H) , 6.14 (br s, 2H) , 6.07 (s, 1H) , 3.95 (t, J ＝ 5.2 Hz, 2H) , 3.67 (t, J ＝ 5.2 Hz, 2H) , 3.36 (s, 3H) , 3.21 (s, 3H) , 2.78 (s, 3H) , 2.36 (s, 3H) . MS: M/e 379 (M+1) ⁺.

Compound B16:

4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (6- (piperazin-1-yl) pyridin-2-yl) pyrimidine-5-carbonitrile

A mixture of

4-amino-6- (6-bromopyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile (56 mg, 0.15 mmol) , piperazine (39 mg, 0.45 mmol) and DIPEA (40 mg, 0.31 mmol) in NMP (1 mL) was heated at 110 ℃ for 12 hrs. The mixture was diluted with EtOAc (10 mL) , washed with brine (10 mL x 3) , dried over anhydrous Na₂SO₄, and concentrated. The resulted residue was purified by column chromatography eluted with DCM/MeOH (100: 1) to give the title product (6 mg, 11％) as a yellow solid. ¹H NMR (400 MHz, DMSO-d6) δ 8.48 (br s, 1H) , 7.84 –7.80 (m, 1H) , 7.60 (d, J ＝7.4 Hz, 1H) , 7.14 (d, J ＝ 8.8 Hz, 1H) , 6.18 (s, 1H) , 3.82 (s, 4H) , 3.07 (s, 4H) , 2.65 (s, 3H) , 2.19 (s, 3H) . MS: M/e 376 (M+1) ⁺.

Compound B17:

4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (6- (4-methylpiperazin-1-yl) pyridin-2-yl) pyrimidine-5-carbonitrile

A mixture of

4-amino-6- (6-bromopyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile (56 mg, 0.15 mmol) , 1-methylpiperazine (45 mg, 0.45 mmol) and DIPEA (40 mg, 0.31 mmol) in NMP (1 mL) was heated at 110 ℃ for 12 hrs. The mixture was diluted with EtOAc (10 mL) , washed with brine (10 mL x 3) , dried over anhydrous Na₂SO₄, and concentrated. The resulted residue was purified by column chromatography eluted with DCM/MeOH (100: 1) to give the title product (16 mg, 27％) as a yellow solid. ¹H NMR (400 MHz, DMSO-d6) δ 8.48 (br s, 1H) , 7.84 –7.80 (m, 1H) , 7.60 (d, J ＝ 7.4 Hz, 1H) , 7.14 (d, J ＝ 8.8 Hz, 1H) , 6.18 (s, 1H) , 3.82 (s, 4H) , 3.07 (s, 4H) , 2.65 (s, 3H) , 2.19 (s, 3H) . MS: M/e 390 (M+1) ⁺.

Compound B18:

4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (6- (4-ethylpiperazin-1-yl) pyridin-2-yl) pyrimidine-5-carbonitrile

A mixture

of4-amino-6- (6-bromopyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile (56 mg, 0.15 mmol) , 1-ethylpiperazine (45 mg, 0.45 mmol) and DIPEA (40 mg, 0.31 mmol) in NMP (1 mL) was heated at 110 ℃ for 12 hrs. The mixture was diluted with EtOAc (10 mL) , washed with brine (10 mL x 3) , dried over anhydrous Na₂SO₄, and concentrated. The resulted residue was purified by column chromatography eluted with DCM/MeOH (100: 1) to give the title product (19 mg, 31％) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 7.71 –7.60 (m, 2H) , 6.82 (s, 1H) , 6.08 (s, 1H) , 6.07 (s, 2H) , 3.79 (t, J ＝ 4.6 Hz, 4H) , 2.78 (s, 3H) , 2.61 (t, J ＝ 4.6 Hz, 4H) , 2.49 (q, J ＝ 7.2 Hz, 2H) , 2.36 (s, 3H) , 1.15 (t, J ＝ 7.2 Hz, 3H) . MS: M/e 404 (M+1) ⁺.

Compound B19:

4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (6- (4-isopropylpiperazin-1-yl) pyridin-2-yl) pyrimidine-5-carbonitrile

A mixture of

4-amino-6- (6-bromopyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile (56 mg, 0.15 mmol) , 1-isopropylpiperazine (58 mg, 0.45 mmol) and DIPEA (40 mg, 0.31 mmol) in NMP (1 mL) was heated at 110 ℃ for 12 hrs. The mixture was diluted with EtOAc (10 mL) , washed with brine (10 mL x 3) , dried over anhydrous Na₂SO₄, and concentrated. The resulted residue was purified by column chromatography eluted with DCM/MeOH (100: 1) to give the title product (22 mg, 35％) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 7.70 –7.58 (m, 2H) , 6.82 (dd, J ＝ 5.6, 2.8 Hz, 1H) , 6.08 (s, 3H) , 3.77 (t, J ＝ 4.4 Hz, 4H) , 2.78 (s, 3H) , 2.76 –2.72 (m, 1H) , 2.68 (t, J ＝ 4.4 Hz, 4H) , 2.36 (s, 3H) , 1.10 (d, J ＝ 6.4 Hz, 6H) . MS: M/e 418 (M+1) ⁺.

Compound B20:

4-amino-6- (6- (4-cyclopropylpiperazin-1-yl) pyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile

Step A: tert-butyl 4-cyclopropylpiperazine-1-carboxylate

To a mixture of (1-ethoxycyclopropoxy) trimethylsilane (1.74 g, 10.0 mmol) , tert-butyl piperazine-1-carboxylate (930 mg, 5.00 mmol) , and AcOH (450 mg, 7.50 mmol) in a mixed solvent (THF/MeOH, 1: 1, 12 mL) was added NaBH₃CN (473 mg, 7.50 mmol) . After heating at 60℃ for 5 hrs, the mixture was added of H₂O (5 mL) , stirred for 5 minutes. Aqueous solution of NaOH (1N, 5 mL) was added and the resulted mixture was extracted with DCM (10 mL x 3) . The combined extracts was dried and concentrated to give the title product (1.11 g, 50％) as a white solid which was used for the next step directly. ¹H NMR (400 MHz, CDCl₃) δ3.39 (t, J ＝ 4.4 Hz, 4H) , 2.55 (t, J ＝ 4.4 Hz, 4H) , 1.67 –1.54 (m, 1H) , 1.46 (s, 9H) , 0.48 –0.47 (m, 2H) , 0.44 –0.42 (m, 2H) . MS: M/e 227 (M+1) ⁺.

Step B: 1-cyclopropylpiperazine hydrochloride

To a solution of the product from Step A (1.11 g, 5.00 mL) in EA (5 mL) was added HCl/EA (10 mL) . After stirring at room temperature for 6 hrs, the mixture was concentrated to dryness to give the title product (1.14 g) as a white solid which was used for the next step directly.

Step C:

A mixture

of4-amino-6- (6-bromopyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile (37 mg, 0.10 mmol) , the product from Step B (49 mg, 0.30 mmol) and DIPEA (60 mg, 0.50 mmol) in NMP (1 mL) was heated at 110 ℃ for 12 hrs. The mixture was diluted with EtOAc (10 mL) , washed with brine (10 mL x 3) , dried over anhydrous Na₂SO₄and concentrated. The resulted residue was purified by column chromatography eluted with DCM/MeOH (100: 1) to give the title product (18 mg, 43％) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 7.70 –7.59 (m, 2H) , 6.82 (dd, J ＝6.8, 2.4 Hz, 1H) , 6.08 (s, 3H) , 3.73 (t, J ＝ 4.8 Hz, 4H) , 2.78 (s, 3H) , 2.76 (t, J ＝ 4.8 Hz, 4H) , 2.36 (s, 3H) , 1.70 –1.63 (m, 1H) , 0.49 –0.48 (m, 4H) . MS: M/e 416 (M+1) ⁺.

Compound B21:

4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (6- (4- (2-hydroxyethyl) piperazin-1-yl) pyridin-2-yl) pyrimidine-5-carbonitrile

A mixture

of4-amino-6- (6-bromopyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile (56 mg, 0.15 mmol) , 2- (piperazin-1-yl) ethan-1-ol (59 mg, 0.45 mmol) and DIPEA (40 mg, 0.31 mmol) in NMP (1 mL) was heated at 110 ℃ for 12 hrs. The mixture was diluted with EtOAc (10 mL) , washed with brine (10 mL x 3) , dried over anhydrous Na₂SO₄, and concentrated. The resulted residue was purified by column chromatography eluted with DCM/MeOH (100: 1) to give the title product (20 mg, 32％) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 7.71 –7.60 (m, 2H) , 6.82 (dd, J ＝ 5.6, 2.8 Hz, 1H) , 6.09 (s, 3H) , 3.78 (s, 4H) , 3.68 (s, 2H) , 2.78 (s, 3H) , 2.68 (s, 4H) , 2.63 –2.62 (m, 2H) , 2.36 (s, 3H) . MS: M/e 420 (M+1) ⁺.

Compound

B22: 4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (6- (4- (2-methoxyethyl) piperazin-1-yl) pyridin-2-yl) pyrimidine-5-carbonitrile

A mixture

of4-amino-6- (6-bromopyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile (56 mg, 0.15 mmol) , 1- (2-methoxyethyl) piperazine (65 mg, 0.45 mmol) and DIPEA (40 mg, 0.31 mmol) in NMP (1 mL) was heated at 110 ℃ for 12 hrs. The mixture was diluted with EtOAc (10 mL) , washed with brine (10 mL x 3) , dried over anhydrous Na₂SO₄, and concentrated. The resulted residue was purified by column chromatography eluted with DCM/MeOH (100: 1) to give the title product (26 mg, 40％) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 7.70 –7.58 (m, 2H) , 6.83 (dd, J ＝ 5.6, 2.8 Hz, 1H) , 6.11 (br s, 2H) , 6.09 (s, 1H) , 3.80 (d, J ＝ 4.4 Hz, 4H) , 3.58 (t, J ＝ 5.6 Hz, 2H) , 3.39 (s, 3H) , 2.78 (s, 3H) , 2.71 –2.57 (m, 6H) , 2.37 (s, 3H) . MS: M/e 434 (M+1) ⁺.

Compound

B23: 4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (3-methoxyphenyl) pyrimidine-5-carbonitrile

Step A: 4-amino-6- (3-methoxyphenyl) -2- (methylthio) pyrimidine-5-carbonitrile

A mixture of3-methoxybenzaldehyde (360 mg, 2.65 mmol) , methyl carbamimidothioate (720 mg, 5.17 mmol) , malononitrile (300 mg, 4.55 mmol) , and NaOH (120 mg, 3.00 mmol) in anhydrous EtOH (8 mL) was refluxed for 16 hrs. The mixture was cooled. The suspension was filtered and the filter cake was washed with H₂O, dried under lamp to give the title product (280 mg, 39％) as a white solid.

Step B: 4-amino-6- (3-methoxyphenyl) -2- (methylsulfonyl) pyrimidine-5-carbonitrile

To a stirred solution of the product from Step A (100 mg, 0.37 mmol) in a mixed solvent (THF/H₂O, 10: 1, 7 mL) was added oxone (250 mg, 0.81 mmol) at room temperature. After stirring for 4 hrs, the mixture was dried with anhydrous Na₂SO₄ (5.0 g) , filtered. The filtrate was concentrated to give the title product (93 mg, 83％) as a yellow solid which was used for the next step directly.

Step

C: 4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (3-methoxyphenyl) pyrimidine-5-carbonitrile

To a stirred solution of the product from Step B (93 mg, 0.31 mmol) in EtOH (3 mL) was added hydrazine hydrate (66 mg, 85％, 1.22 mmol) . The mixture was stirred at 80 ℃ for 2 hrs and concentrated. The resulted residue was added EtOH (3 mL) and followed by pentane-2, 4-dione (36 mg, 0.36 mmol) . After being refluxed for 4 hrs, the mixture was cooled, filtered. The filter cake was washed with EtOH to give the title product (35 mg, 35％) as a white solid. ¹H NMR (400 MHz, CDCl₃) δ7.67 (d, J ＝ 7.6 Hz, 1H) , 7.59 (s, 1H) , 7.46 (dd, J ＝ 8.0, 7.6 Hz, 1H) , 7.13 (d, J ＝ 8.4 Hz, 1H) , 6.09 (s, 1H) , 6.00 (br s, 2H) , 3.90 (s, 3H) , 2.74 (s, 3H) , 2.36 (s, 3H) . MS: M/e 321 (M+1) ⁺.

Compound

B24: 4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (2-methylpyridin-4-yl) pyrimidine-5-carbonitrile

Step A: 2-methylisonicotinaldehyde

To a stirred solution of 4-bromo-2-methylpyridine (1.00 g, 5.81 mmol) in anhydrous diethyl ether (70 mL) was added n-BuLi (2.5 M, 2.56 mL, 6.39 mmol) at -78 ℃ under N₂. After stirring at -78 ℃ for 15 min, DMF (0.54 mL, 6.92 mmol) was added slowly. The mixture was stirred for another 30 min and warmed to 0 ℃. The mixture was quenched with 20 mL of aq. NaHCO₃, extracted with EA (50 mL x 2) . The combined organics was washed with brine (50 mL x 2) , dried over anhydrous Na₂SO₄, filtered and concentrated. The resulted residue was purified by column chromatography eluted with PE/EA (5: 1) to give the title product (210 mg, 30％) as a light yellow solid. ¹H NMR (400 MHz, CDCl₃) δ10.05 (s, 1H) , 8.76 (d, J ＝4.8 Hz, 1H) , 7.56 (s, 1H) , 7.51 (d, J ＝4.8 Hz, 1H) , 2.68 (s, 3H) .

A mixture ofthe product from step A (120 mg, 1 mmol) , methyl carbamimidothioate (139 mg, 1.00 mmol) , malononitrile (99 mg, 1.50 mmol) , and NaOH (40 mg, 1.00 mmol) in anhydrous EtOH (10 mL) was refluxed for 46 hrs. The mixture was cooled. The suspension was filtered and the filter cake was washed with H₂O, dried under lamp to give the title product (90 mg, 35％) as a white solid. ¹H NMR (400 MHz, DMSO-d6) δ 8.63 (d, J ＝ 5.2 Hz, 1H) , 7.61 (s, 1H) , 7.57 (d, J ＝ 5.0 Hz, 1H) , 2.56 (s, 3H) . MS: M/e 258 (M+1) ⁺.

Step C:

4-amino-6- (2-methylpyridin-4-yl) -2- (methylsulfonyl) pyrimidine-5-carbonitrile

To a stirred solution of the product from Step B (90 mg, 0.35 mmol) in a mixed solvent (THF/H₂O, 10/1, 4 mL) was added oxone (270 mg, 0.88 mmol) at room temperature. After stirring for 4 hrs, the mixture was dried with anhydrous Na₂SO₄ (5.0 g) , filtered. The filtrate was concentrated to give the title product (60 mg) as a yellow solid which was used for the next step directly.

Step

D: 4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (2-methylpyridin-4-yl) pyrimidine-5-carbonitrile

To a stirred solution of the product from Step C (60 mg, 0.35 mmol) in EtOH (5 mL) was added hydrazine hydrate (50 mg, 85％, 0.85 mmol) . The mixture was stirred at 80 ℃ for 2 hrs and concentrated. The resulted residue was added EtOH (5 mL) and followed by pentane-2, 4-dione (85 mg, 0.85 mmol) . After being refluxed for 4 hrs, the mixture was cooled and concentrated. The resulted residue was purified by column chromatography eluted with (DCM/MeOH ＝ 50: 1) to give the title product (40 mg, 37％) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 8.72 (d, J ＝ 5.2 Hz, 1H) , 7.74 (s, 1H) , 7.67 (d, J ＝ 5.0 Hz, 1H) , 6.15 (s, 1H) , 6.09 (s, 1H) , 2.70 (s, 3H) , 2.69 (s, 3H) , 2.36 (s, 3H) . MS: M/e 306 (M+1) ⁺.

Compound

B25: 4-amino-6- (5-methoxypyridin-3-yl) -2-phenylpyrimidine-5-carbonitrile

Step A: 5-methoxynicotinaldehyde

To a stirred solution of 3-bromo-5-methoxypyridine (1.00 g, 5.32 mmol) in a mixed solvent (THF/hexane＝1: 1, 20 mL) was added n-BuLi (2.5 M, 2.4 mL) at -78 ℃ under N₂. After stirring at -78 ℃ for 15 min, DMF (0.54 mL, 6.92 mmol) was added slowly. The mixture was stirred for another 30 min and warmed to 0 ℃. The mixture was quenched with 20 mL of aq. NaHCO₃, extracted with EA (50 mL x 2) . The combined organics was washed with brine (50 mL x 2) , dried over anhydrous Na₂SO₄, filtered and concentrated. The resulted residue was purified by column chromatography eluted with PE/EA (5: 1) to give the title product (360 mg, 49％) as a light yellow solid.

Step B: 4-amino-6- (5-methoxypyridin-3-yl) -2-phenylpyrimidine-5-carbonitrile

A mixture ofthe product from step A (15 mg, 0.11 mmol) , benzimidamide hydrochloride (17 mg, 0.11 mmol) , malononitrile (11 mg, 0.17 mmol) , and NaOH (4 mg, 0.10 mmol) in anhydrous EtOH (5 mL) was refluxed for 16 hrs. The mixture was cooled. The suspension was filtered and the filter cake was washed with H₂O and EtOH, dried under lamp to give the title product (2 mg, 6％) as a white solid. ¹H NMR (400 MHz, CDCl₃) δ 8.96 (s, 1H) , 8.50 (m, 3H) , 7.91 (s, 1H) , 7.56 –7.49 (m, 3H) , 5.79 (s, 2H) , 3.98 (s, 3H) . MS: M/e 304 (M+1) ⁺.

Compound

B26: 4-amino-6- (5-methoxypyridin-3-yl) -2- (pyridin-3-yl) pyrimidine-5-carbonitrile

A mixture of5-methoxynicotinaldehyde (41 mg, 0. 30 mmol) , nicotinimidamide hydrochloride (47 mg, 0.30 mmol) , malononitrile (30 mg, 0.45 mmol) , and NaOH (12 mg, 0.30 mmol) in anhydrous EtOH (5 mL) was refluxed for 16 hrs. The mixture was cooled. The suspension was filtered and the filter cake was washed with H₂O and EtOH, dried under lamp to give the title product (25 mg, 27％) as a white solid. ¹H NMR (400 MHz, DMSO-d6) δ 9.50 (d, J ＝ 1.4 Hz, 1H) , 8.78 –8.71 (m, 2H) , 8.66 (d, J ＝ 8.0 Hz, 1H) , 8.52 (d, J ＝ 2.8 Hz, 1H) , 7.92 (s, 1H) , 7.58 (dd, J ＝8.0, 5.0 Hz, 1H) , 3.94 (s, 3H) . MS: M/e 305 (M+1) ⁺.

Compound

B27: 4-amino-6- (5-methoxypyridin-3-yl) -2- (pyridin-4-yl) pyrimidine-5-carbonitrile

A mixture of5-methoxynicotinaldehyde (41 mg, 0.30 mmol) , isonicotinimidamide hydrochloride (47 mg, 0.30 mmol) , malononitrile (30 mg, 0.45 mmol) , and NaOH (12 mg, 0.30 mmol) in anhydrous EtOH (5 mL) was refluxed for 16 hrs. The mixture was cooled. The suspension was filtered and the filter cake was washed with H₂O and EtOH, dried under lamp to give the title product (28 mg, 31％) as a white solid. ¹H NMR (400 MHz, DMSO-d6) δ 8.79 (d, J ＝ 5.8 Hz, 2H) , 8.73 (s, 1H) , 8.52 (d, J ＝ 2.8 Hz, 1H) , 8.23 (d, J ＝ 5.8 Hz, 2H) , 7.92 (s, 1H) , 3.94 (s, 3H) . MS: M/e 305 (M+1) ⁺.

Compound

B28: 4-amino-6- (5-methoxypyridin-3-yl) -2- (1H-pyrazol-1-yl) pyrimidine-5-carbonitrile

Step A: 4-amino-6- (5-methoxypyridin-3-yl) -2- (methylthio) pyrimidine-5-carbonitrile

A mixture of5-methoxynicotinaldehyde (137 mg, 1.0 mmol) , methyl carbamimidothioate (139 mg, 1.00 mmol) , malononitrile (99 mg, 1.50 mmol) , and NaOH (40 mg, 1.00 mmol) in anhydrous EtOH (10 mL) was refluxed for 16 hrs. The mixture was cooled. The suspension was filtered and the filter cake was washed with H₂O, dried under lamp to give the title product (37 mg, 14％) as a white solid.

Step B: 4-amino-6- (5-methoxypyridin-3-yl) -2- (methylsulfonyl) pyrimidine

-5-carbonitrile

To a stirred solution of the product from Step A (37 mg, 0.14 mmol) in a mixed solvent (THF/H₂O, 10: 1, 4 mL) was added oxone (104 mg, 0.34 mmol) at room temperature. After stirring at for 4 hrs, the mixture was dried with anhydrous Na₂SO₄ (5.0 g) , filtered. The filtrate was concentrated to give the title product (16 mg) as a yellow solid which was used for the next step directly.

Step

C: 4-amino-6- (5-methoxypyridin-3-yl) -2- (1H-pyrazol-1-yl) pyrimidine-5-carbonitrile

To a stirred solution of the product from Step B (16 mg, 0.052 mmol) and 1H-pyrazole (4 mg, 0.059 mmol) in THF (2 mL, anhydrous) was added NaH (2 mg, 80％, 0.067 mmol) at 0 ℃. After stirring at room temperature for 6 hrs, the mixture was quenched with H₂O, extracted with EA. The combined extracts was dried and concentrated. The resulted residue was purified by column chromatography eluted with DCM/MeOH (100: 1) to give the title product (3 mg, 19％) as a white solid. ¹H NMR (400 MHz, CDCl₃) δ 8.89 (s, 1H) , 8.65 (s, 1H) , 8.52 (s, 1H) , 7.87 (s, 1H) , 7.86 (s, 1H) , 6.54 (s, 1H) , 6.14 (br s, 2H) , 3.97 (s, 3H) . MS: M/e 294 (M+1) ⁺.

Compound

B29: 4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (5-methoxypyridin-3-yl) pyrimidine-5-carbonitrile

To a mixture

of4-amino-6- (5-methoxypyridin-3-yl) -2- (methylsulfonyl) pyrimidine-5-carbonitrile (45 mg, 0.15 mmol) and 3, 5-dimethyl-1H-pyrazole (17 mg, 0.18 mmol) in DMF (1 mL) was added NaH (18 mg, 80％, 0.6 mmol) at 0 ℃. After stirring at room temperature for 6 hrs, the mixture was poured into 5 mL of H₂O. The resulted suspension was filtered and the filter cake was washed with H₂O, dried to give the title product (13 mg, 28％) as a gray solid. ¹H NMR (400 MHz, CDCl₃) δ 8.87 (s, 1H) , 8.50 (d, J ＝ 2.8 Hz, 1H) , 7.85 (s, 1H) , 6.14 (s, 2H) , 6.09 (s, 1H) , 3.95 (s, 3H) , 2.72 (s, 3H) , 2.36 (s, 3H) . MS: M/e 322 (M+1) ⁺.

Compound

B30: 2- (3, 5-dimethyl-1H-pyrazol-1-yl) -5-fluoro-6- (5-methoxypyridin-3-yl) pyrimidin-4-amine

Step A: 6-chloro-5-fluoro-2- (methylthio) pyrimidin-4-amine

To a solution of 4, 6-dichloro-5-fluoro-2- (methylthio) pyrimidine (1.5 g, 7.04 mmol) was added ammonia hydrate (4.79 g, 25％, 70.4 mmol) . After stirring at 60 ℃ for 3 hrs the mixture was cooled. 50 mL of H₂O was added. The mixture was extracted with EA (50 mL x 3) . The combined extracts was dried with anhydrous Na₂SO₄, filtered and concentrated to give the title product (1.38 g, 100％) as a yellow solid.

Step B: 6-chloro-5-fluoro-2- (methylsulfonyl) pyrimidin-4-amine

To a stirred solution of the product from Step A (1.46 g, 7.53 mmol) in DCM (50 mL) was added mCPBA (3.25 g, 18.81 mmol) in portions at 0 ℃ and the mixture was stirred for 16 hrs. (Overnight) . A mixed solution of sodium thiosulphate (1.2 g, 7.59 mmol) and sodium bicarbonate (2.0 g, 23.81 mmol) in H₂O (50 mL) was added and the resulted mixture was stirred for another 2 hrs, filtered. A white solid was collected and dried. The filtrate was extracted with EA (50 mL x 3) . The combined extracts was dried, concentrated and the resulted residue was purified by column chromatography eluted with (From PE/EA＝3: 1 to EA＝100％) to give the title product, which was combined with the white solid to give the title product (1.6 g, 94％) as a white solid.

Step C: 6-chloro-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -5-fluoropyrimidin-4-amine

To a mixture ofthe product from Step B (760 mg, 3.36 mmol) and 3, 5-dimethyl-1H-pyrazole (387 mg, 4.04 mmol) in anhydrous THF was added NaH (404 mg, 80％, 13.45 mmol) at 0 ℃. After stirring at room temperature for 3 hrs, the mixture was poured into H₂O and extracted with EA (50 mL x 3) . The combined extracts was dried with anhydrous Na₂SO₄, filtered and concentrated. The resulted residue was purified by column chromatography eluted with (DCM/MeOH＝50: 1) to give the title product (525 mg, 64％) as a white solid. ¹H NMR (400 MHz, CDCl₃) δ6.00 (s, 1H) , 5.54 (s, 2H) , 2.61 (s, 3H) , 2.31 (s, 3H) . MS: M/e 242 (M+1) ⁺.

Step D: potassium trifluoro (5-methoxypyridin-3-yl) borate

Amixed solution of 3-bromo-5-methoxypyridine (2.2 g, 97％, 11.4 mmol) and triisopropyl borate (2.6 g, 98％, 13.6 mmol) in a mixed solvent (THF/Toluene, 24mL/6mL) was stirred at -40 ℃ under N₂ for 10 min. n-BuLi (5.4 mL, 13.6 mmol) was added in drops. After being stirred at this temperature for 2 hrs, the mixture was warmed to room temperature and a solution of KHF₂ (2.7 g, 34.0 mmol) in H₂O was added and the mixture was stirred for another 4 hrs. The mixture was concentrated and the residue was dissolved in MeOH (100 mL) . The solution was filtered and the filtrate was concentrated. The resulted residue was washed with diethylether (50 mL x 3) , and the diethylether layers were removed. The resulted oil was dissolved in acetone (100 mL) , filtered, and the filtrate was concentrated to give the title product (1.8 g, 74％) as a brown oil.

Step

E: 2- (3, 5-dimethyl-1H-pyrazol-1-yl) -5-fluoro-6- (5-methoxypyridin-3-yl) pyrimidin-4-amine

Amixture of the product from Step C (550 mg, 2.27 mmol) , the product from Step D (733 mg, 3.41 mmol) , Pd (PPh₃) ₄ (263 mg, 0.23 mmol) and K₂CO₃ (1.25 g, 9.09 mmol) in a mixed solvent (Dioxane/H₂O, 20mL/4mL) was refluxed for 12 hrs under N₂. The mixture was cooled to room temperature and filtered through a celite pad. 100 mL of H₂O was added into the filtrate and a yellow solid precipitated. The resulted suspension was filtered and the filter cake was dried and purified by column chromatography eluted with (DCM/MeOH＝50: 1) to give the title product (380 mg, 53％) as a yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 8.93 (s, 1H) , 8.42 (d, J ＝ 2.4 Hz, 1H) , 7.92 (s, 1H) , 6.04 (s, 1H) , 5.57 (s, 2H) , 3.94 (s, 3H) , 2.71 (s, 3H) , 2.34 (s, 3H) . MS: M/e 315 (M+1) ⁺.

Compound B31:

(R) -4-amino-6- (6- (4- (2-cyanoethyl) -3-methylpiperazin-1-yl) pyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile

Step A: (R) -3- (2-methylpiperazin-1-yl) propanenitrile

Amixture of (R) -tert-butyl 3-methylpiperazine-1-carboxylate (200 mg, 1.00 mmol) , acrylonitrile (106 mg, 2.00 mol) and TEA (101 mg, 1.00 mmol) in 5 mL MeOH was stirred at room temperature overnight. After that, the mixture was portioned between ethyl acetate and water. The organic phase was concentrated. The residue was redissolved in 5 mL CHCl₂/TFA (1 : 1) . The mixture was stirred at room temperature for 4 h. Then the solvent was removed by vacuum to give a light-brown oil without purification for next step (150 mg, crude) .

Step B:

(R) -4-amino-6- (6- (4- (2-cyanoethyl) -3-methylpiperazin-1-yl) pyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile (2)

A mixture of

4-amino-6- (6-bromopyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile (144 mg, 0.45 mmol) , (R) -3- (2-methylpiperazin-1-yl) propanenitrile (150 mg, 1.00 mmol) , DIPEA (650 mg, 5.00 mmol) in 6 mL NMP was stirred at 110℃ for 36 h. After cooling to room temperature, the mixture was diluted with ethyl acetate. The organic layer was washed with brine and concentrated. The residue was purified by column chromatography (1％MeOH/CHCl₂) to give the desired compound (12 mg, 6％) . ¹H NMR (400 MHz, CDCl₃) δ 7.66-7.64 (m, 2H) , 6.86–6.76 (m, 1H) , 6.08 (s, 1H) , 6.03 (br s, 2H) , 4.26 (d, J ＝ 12.4 Hz, 1H) , 4.18 (d, J ＝ 12.4 Hz, 1H) , 3.36-3.33 (m, 1H) , 3.13-3.06 (m, 1H) , 2.99-2.93 (m, 2H) , 2.77 (s, 3H) , 2.76-2.74 (m, 1H) , 2.64-2.62 (m, 1H) , 2.58–2.48 (m, 3H) , 2.36 (s, 3H) , 1.19 (d, J ＝ 6.0 Hz, 3H) . MS: M/e 443 (M+1) ⁺.

Compound B32:

(S) -4-amino-6- (6- (4- (2-cyanoethyl) -3-methylpiperazin-1-yl) pyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile

Step A: (S) -3- (2-methylpiperazin-1-yl) propanenitrile (3-1)

Amixture of (S) -tert-butyl 3-methylpiperazine-1-carboxylate (200 mg, 1.00 mmol) , acrylonitrile (106 mg, 2.00 mol) and TEA (101 mg, 1.00 mmol) in 5 mL MeOH was stirred at room temperature overnight. After that, the mixture was portioned between ethyl acetate and water. The organic phase was concentrated. The residue was redissolved in 5 mL CHCl₂/TFA (1 : 1) . The mixture was stirred at room temperature for 4 h. Then the solvent was removed by vacuum to give a light-brown oil without purification for next step (130 mg, crude) .

Step B:

A mixture of

4-amino-6- (6-bromopyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile (130 mg, 0.35 mmol) , (S) -3- (2-methylpiperazin-1-yl) propanenitrile (130 mg, 0.87 mmol) , DIPEA (650 mg, 5.00 mmol) in 6 mL NMP was stirred at 110℃ for 36 h. After cooled to room temperature, the mixture was diluted with ethyl acetate. The organic layer was washed with brine and concentrated. The residue was purified by column chromatography (1％MeOH/CHCl₂) to give the desired compound (10 mg, 6％) . ¹H NMR (400 MHz, CDCl₃) δ 7.65-7.63 (m, 2H) , 6.86 –6.74 (m, 1H) , 6.08 (s, 1H) , 6.03 (br. s, 2H) , 4.26 (d, J ＝ 12.8 Hz, 1H) , 4.17 (d, J ＝ 12.8 Hz, 1H) , 3.36-3.33 (m, 1H) , 3.13-3.06 (m, 1H) , 3.04–2.90 (m, 2H) , 2.77 (s, 3H) , 2.76-2.74 (m, 1H) , 2.68-2.63 (m, 1H) , 2.59–2.46 (m, 3H) , 2.36 (s, 3H) , 1.19 (d, J ＝ 6.0 Hz, 3H) . MS: M/e 443 (M+1) ⁺.

Compound B33:

(S) -4-amino-6- (6- (4- (2-cyanoethyl) -2-methylpiperazin-1-yl) pyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile

Step A:

(S) -4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (6- (2-methylpiperazin-1-yl) pyridin-2-yl) pyrimidine-5-carbonitrile

A mixture of

4-amino-6- (6-bromopyridin-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile (370 mg, 1.00 mmol) , (S) -tert-butyl 3-methylpiperazine-1-carboxylate (1.0 g, 5.00 mmol) , DIPEA (650 mg, 5.00 mmol) in 10 mL NMP was stirred at 110℃ for 36 h. After cooling to room temperature, the mixture was diluted with ethyl acetate. The organic layer was washed with brine and concentrated. The residue was purified by column chromatography (4％MeOH/CHCl₂) to give the desired compound (20 mg, 5％) .

Step B:

(S) -4-amino-6- (6- (4- (2-cyanoethyl) -2-methylpiperazin-1-yl) pyridine-2-yl) -2- (3, 5-dimethyl-1H-pyrazol-1-yl) pyrimidine-5-carbonitrile

A mixture of

(S) -4-amino-2- (3, 5-dimethyl-1H-pyrazol-1-yl) -6- (6- (2-methylpiperazin-1-yl) pyridin-2-yl) pyrimidin e-5-carbonitrile (18 mg, 0.051 mmol) , acrylonitrile (6 mg, 0.11 mol) and TEA (5 mg, 0.049 mmol) in 1 mL MeOH was stirred at room temperature overnight. Then the solvent was removed and the residue was purified by column chromatography (1％MeOH/CHCl₂) to give final compound (2 mg, 9％). ¹H NMR (400 MHz, CDCl₃) δ 7.65-7.63 (m, 2H) , 6.85–6.67 (m, 1H) , 6.08 (s, 1H) , 6.02 (br s, 2H) , 4.74-4.63 (m, 1H) , 4.30 (d, J ＝ 12.8 Hz, 1H) , 3.29 (t, J ＝ 12.8 Hz, 1H) , 2.99 (d, J ＝ 11.0 Hz, 1H) , 2.83 (d, J ＝ 11.0 Hz, 1H) , 2.77 (s, 3H) , 2.75–2.63 (m, 2H) , 2.56 (t, J ＝ 6.8 Hz, 2H) , 2.48-2.44 (m, 1H) , 2.36 (s, 3H) , 2.33–2.24 (m, 1H) , 1.32 (d, J ＝ 6.8 Hz, 3H) . MS: M/e 443 (M+1) ⁺.

BIOLOGICAL ACTIVITIES:

1. The binding ability of compounds to A2Aadenosine receptor assay

The binding ability of the compounds in Table 1 to A2A adenosine receptor was determined, comprising the following steps:

1) . Membrane Preparation

HEK293 (G418 resistant) cells stably expressing adenosine A2A were collected and dissolvedinto lysis buffer (5 mM Tris base, pH 7.4, EDTA·Na 25 mM, EGTA 5 mM, PMSF 1: 1000) , and then lysed on ice for 30 min. The lysate was passed through needle (1 mL needle) on ice bath for 15 times, and then high speed centrifugation (40000 r/min, 4℃, 20 min) to give crude HEK293/A2A cell membrane. The resulting crude membrane was dissolved in reaction buffer (50mM Tris, pH 7.4, 2mM MgCl2) , passed through needle (1 mL needle) on ice bath for 15 times, and then high speed centrifugation (40000 r/min, 4℃, 20 min) to give HEK293/A2A membrane protein. The membrane protein was dissolved in 500 μL reaction buffer and passed through needle (1 mL needle) on ice bath for 10 times. Protein concentration was measured by BCA assay, and the protein was stored in a refrigerator at -80℃.

2) . Binding Assay

To the solution of the membrane protein was added 1 U/mL adenosine deaminase. Non-specific binding was measures in the presence of NECA (10 μM) . In the presence of different concentrations of competitive ligands, membrane protein (50 μg) and 0.1 nM [3H] ZM241385 (50.00 Ci/mmol) were incubated in water bath at 37℃ for 30 min. Thereactionwasterminatedin anice-waterbath. The bound ligands and free ligands were separated by GF/B glass fiber filter papers through vacuum filtration on a 12-well Millipore cell sample collector, and then were washed with ice cold 50 mM Tris-HCl for 3 times. The membrane was oven-dried and placed in EP tube, to which 540 μL scintillation solutions were added. The binding of the radioactive ligands was measured by Beckman LS-6500 Liquid Scintillation Counter. The percentage competitive inhibition rate of each Examplegainst the binding of isotope with the protein receptor was calculated according to the following formula, where cpm is the reading value of the radioactive ligand measured by the assay:

Inhibition rate (I％) ＝ (Total binding tube cpm-compound cpm) / (Total binding tube cpm-non-specific binding tube cpm) ×100％

The Ki value of a test compound was calculatedfrom a plot of the concentration of the Examples X-axis versus the competitive inhibition rate of each Examplegainst the binding of isotope with the protein receptor as Y-axis. The smaller the Ki value, the better the binding ability of the compound to A2A adenosine receptor.

The A2A binding ability of the compounds synthesized in above Examples were shown in Table 2:

Table 2

Each of the above compounds has shown good inhibition rate. The aminopyrimidine heterocyclic compounds of the Examples areeffective adenosine receptor antagonists, have a strong binging to the receptor, and effectively block adenosine receptor. The aminopyrimidine heterocyclic compounds disclosed herein can be used for the treatment or prevention of disorders caused by abnormal level of adenosine.

2. The function assay

The effect of some compounds in Table 1 on the function of the T-cell surface A2A adenosine receptor-mediated A2A receptor binding dependent signaling pathway was determined, comprising the following steps:

Lymphocytes isolated from the spleen of C57/BL6 mice were placed in 96-well plate with 5×10⁵ cells each well. Mouse spleen cells would produce IFN-γ under the induction of 0.1 μg/mL CD3 monoclonal antibody (mAb) . After the addition of CD3 mAb to mouse spleen cells, the addition of 100 nM A2A receptor agonist CGS21680 could inhibit the production of IFN-γ. In order to test the inhibitory activity of the synthesized compounds on A2A receptor in lymphocytes, different concentrations of test compounds taken together with 100 nM A2A receptor agonist CGS21680 were added to mouse spleen cells to which CD3 mAb had been added. After 24 h, the supernatant was collected to perform enzyme-linked immunosorbent assay (ELISA assay) by using a kit from eBioscience (Cat: #887314) , and determine the concentration of IFN-γ in the supernatant. The EC50 value of a test compound was calculatedfrom a histogram of the concentration of the Examples X-axis versus the concentration of IFN-γ in the supernatant as Y-axis. The smaller the EC50 value, the better the inhibitory ability of T-cell surface A2A adenosine receptor-mediated A2A receptor binding dependent signaling pathway.

The effect of some compounds in Table 1 on the function of the T-cell surface A2A adenosine receptor-mediated A2A receptor binding dependent signaling pathway was shown in Table 3:

Table 3

Each of the above compounds has shown good inhibitory effects on the T-cell surface A2A adenosine receptor-mediated A2A receptor binding dependent signaling pathway. The aminopyrimidine heterocyclic compounds of the Examples are effective adenosine receptor antagonists, are able to effectively block the adenosine receptor on lymphocyte surface such that the cancer cells fail to escape from immune surveillance. The aminopyrimidine heterocyclic compounds disclosed hereincan thusbe used for the treatment or prevention of cancer.

The present invention can be performed in various manners, where all the embodiments, such as adaptations, variations, modifications, and equivalent arrangements, fall within the protection scope of the present invention.

Claims

An aminopyrimidine heterocyclic compound with adenosine receptor antagonistic activity, comprising a compound of the general formula (I) ,

wherein:

A₁, A₂, A₃, and A₄ are each independently selected from the group consisting of N and CR₆；

R₁ is aryl or 5-6 membered heteroaryl comprising 1-3 heteroatoms independently selected from N, O and S, wherein said aryl or 5-6 membered heteroaryl is unsubstituted or substituted by 1-3 R₇ groups；

R₂ is independently selected from the group consisting of cyano and halo；

R₃ is independently selected from the group consisting of halo, cyano, C_1-6 alkyl, C_3-6 cycloalkyl, C_2-6 alkenyl, C_2-6 alkynyl, C_2-6 ureido, C_2-6 oxoureido, C_1-6 alkoxy, C_1-6 acyl, NR₄R₅ and CONR₄R₅；

R₄ andR₅ are each independently selected from the group consisting of hydrogen, C_1-6 alkyl, C_3-6 cycloalkyl, and C_1-6acyl, wherein said alkyl, cycloalkyl, and acyl may be substituted by 1-3 R₈ groups； or R₄ and R₅, together with the nitrogen atom to which they are attached, may form a saturated heterocycle comprising 1-3 heteroatoms independently selected from the group consisting of N, O and S, and said saturated heterocycle may be substituted by 1-3 R₉ groups, and two R₉ substituents may, together with the atom to which they are attached, also form a ring；

R₆ and R₇ are each independently selected from the group consisting of hydrogen, deuterium, halo, hydroxy, cyano, C_3-6 cycloalkyl, C_1-6 alkoxy, and C_1-6 alkyl unsubstituted or substituted by halo or deuterium；

R₈ and R₉ are each independently selected from the group consisting of hydrogen, deuterium, halo, hydroxy, cyano, morpholino, piperazin-1-yl, C_3-6 cycloalkyl, C_1-6 alkoxy, and C_1-6 alkyl unsubstituted or substituted by halo or deuterium；

or a pharmaceutically acceptable salt thereof.
The Example according to Claim 1, which is a compound of the general formula (II) ,

wherein:

A₁, A₂, A₃, and A₄ are each independently selected from the group consisting of N and CR₆；

R₁ is aryl or 5-6 membered heteroaryl comprising 1-3 heteroatoms independently selected from N, O and S, wherein said aryl or 5-6 membered heteroaryl is unsubstituted or substituted by 1-3 R₇ groups；

R₃ is independently selected from the group consisting of halo, cyano, C_1-6 alkyl, C_3-6 cycloalkyl, C_2-6 alkenyl, C_2-6 alkynyl, C_2-6 ureido, C_2-6 oxoureido, C_1-6 alkoxy, C_1-6 acyl, NR₄R₅ and CONR₄R₅；

R₄ andR₅ are each independently selected from the group consisting of hydrogen, C_1-6 alkyl, C_3-6 cycloalkyl, and C_1-6acyl, wherein said alkyl, cycloalkyl, and acyl may be substituted by 1-3 R₈ groups； or R₄ and R₅, together with the nitrogen atom to which they are attached, may form a saturated heterocycle comprising 1-3 heteroatoms independently selected from the group consisting of N, O and S, and said saturated heterocycle may be substituted by 1-3 R₉ groups, and two R₉ substituents may, together with the atom to which they are attached, also form a ring；

R₆ and R₇ are each independently selected from the group consisting of hydrogen, deuterium, halo, hydroxy, cyano, C_3-6 cycloalkyl, C_1-6 alkoxy, and C_1-6 alkyl unsubstituted or substituted by halo or deuterium；

R₈ and R₉ are each independently selected from the group consisting of hydrogen, deuterium, halo, hydroxy, cyano, morpholino, piperazin-1-yl, C_3-6 cycloalkyl, C_1-6 alkoxy, and C_1-6 alkyl unsubstituted or substituted by halo or deuterium；

or a pharmaceutically acceptable salt thereof.
The Example according to any one of Claims 1-2, wherein R₁ is the following groups,

wherein each of the above groups is unsubstituted or substituted by 1-3 R₇ groups, and R₇ group is as defined in Claim 1.
The Example according to any one of Claims 1-3, which is a compound of the general formula (IIA) ,

wherein:

R₁ is independently selected from
or

A₁, A₂, A₃, and A₄ are each independently selected from the group consisting of N and CR₆；

wherein A₁, A₂, A₃, and A₄ are each CR₆； alternatively, one of A₁, A₂, A₃, and A₄ is N, the others are each CR₆；

R₃ is independently selected from the group consisting of halo, cyano, C_1-6 alkyl, C_3-6 cycloalkyl, C_2-6 alkenyl, C_2-6 alkynyl, C_2-6 ureido, C_2-6 oxoureido, C_1-6 alkoxy, C_1-6 acyl, NR₄R₅ and CONR₄R₅；

R₄ and R₅ are each independently selected from the group consisting of hydrogen, C_1-6 alkyl, C_3-6 cycloalkyl, and C_1-6acyl, wherein said alkyl, cycloalkyl, and acyl may be substituted by 1-3 R₈ groups； or R₄ and R₅, together with the nitrogen atom to which they are attached, may form a saturated heterocycle comprising 1-3 heteroatoms independently selected from the group consisting of N, O and S, and said saturated heterocycle may be substituted by 1-3 R₉ groups, and two R₉ substituents may, together with the atom to which they are attached, also form a ring；

R₆ and R₇ are each independently selected from the group consisting of hydrogen, deuterium, halo, hydroxy, cyano, C_3-6 cycloalkyl, C_1-6 alkoxy, and C_1-6 alkyl unsubstituted or substituted by halo or deuterium；

R₈ and R₉ are each independently selected from the group consisting of hydrogen, deuterium, halo, hydroxy, cyano, morpholino, piperazin-1-yl, C_3-6 cycloalkyl, C_1-6 alkoxy, and C_1-6 alkyl unsubstituted or substituted by halo or deuterium；

R₁₀, R₁₁, and R₁₂ are each independently selected from the group consisting of hydrogen, deuterium, halo, cyano, C_3-6 cycloalkyl, and C_1-6 alkyl unsubstituted or substituted by halo or deuterium；

or a pharmaceutically acceptable salt thereof.
The Example according to any one of Claims 1-4, which is a compound of the general formula (IIB) ,

wherein:

R₁ is independently selected from
or

A₁, A₂, A₃, and A₄ are each independently selected from the group consisting of N and CR₆；

wherein A₁, A₂, A₃, and A₄ are each CR₆； alternatively, one of A₁, A₂, A₃, and A₄ is N, the others are each CR₆；

R₃ is NR₄R₅；

R₄ and R₅ are each independently selected from the group consisting of hydrogen, C_1-6 alkyl, C_3-6 cycloalkyl, and C_1-6 acyl, wherein said alkyl, cycloalkyl, and acyl may be substituted by 1-3 R₈ groups； or R₄ and R₅, together with the nitrogen atom to which they are attached, may form a saturated heterocycle comprising 1-3 heteroatoms independently selected from the group consisting of N, O and S, and said saturated heterocycle may be substituted by 1-3 R₉ groups, and two R₉ substituents may, together with the atom to which they are attached, also form a ring；

R₆ and R₇ are each independently selected from the group consisting of hydrogen, deuterium, halo, hydroxy, cyano, C_3-6 cycloalkyl, C_1-6 alkoxy, and C_1-6 alkyl unsubstituted or substituted by halo or deuterium；

R₈ and R₉ are each independently selected from the group consisting of hydrogen, deuterium, halo, hydroxy, cyano, morpholino, piperazin-1-yl, C_3-6 cycloalkyl, C_1-6 alkoxy, and C_1-6 alkyl unsubstituted or substituted by halo or deuterium；

R₁₀, R₁₁, and R₁₂ are each independently selected from the group consisting of hydrogen, deuterium, halo, cyano, C_3-6 cycloalkyl, and C_1-6 alkyl unsubstituted or substituted by halo or deuterium；

or a pharmaceutically acceptable salt thereof.
The Example according to any one of Claims 1-5, wherein R₃ is selected from the following atom or group:
The Example according to any one of Claims 1-6, which is selected from the following compound, and the pharmaceutically acceptable salts thereof.
A pharmaceutical composition comprising, an aminopyrimidine heterocyclic compound with adenosine receptor antagonistic activity according to any one of Claims 1-7, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier or diluent.
A combination comprisingan aminopyrimidine heterocyclic compound with adenosine receptor antagonistic activity, wherein the combination comprises an aminopyrimidine heterocyclic compound with adenosine receptor antagonistic activity according to any one of Claims 1-7 in combination with one or more of L-DOPA, dopamine agonists, dopamine decarboxylase inhibitors, catechol-O-methyltransferase inhibitors and monoamine oxidase inhibitors, cancer immunotherapy such as cancer vaccines, immune checkpoint inhibitors such as cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) and programmed cell death protein 1 (PD-1) .
Use of an aminopyrimidine heterocyclic compound with adenosine receptor antagonistic activity according to any one of Claims 1-7, the pharmaceutical composition comprising an aminopyrimidine heterocyclic compound with adenosine receptor antagonistic activity according to Claims 8, or the combination comprising an aminopyrimidine heterocyclic compound with adenosine receptor antagonistic activity according to Claim 9 in the manufacture of a medicament for antagonising an adenosine receptor.