EA042196B1 - METHOD FOR OBTAINING SOMATOSTATIN MODULATORS - Google Patents

Description

Related Applications

The present application claims priority under U.S. Provisional Application No. 62/618,538, filed January 17, 2018, which is incorporated herein by reference in its entirety.

The field of technology to which the present invention relates

This document describes compounds that are modulators of somatostatin, methods for preparing such compounds, pharmaceutical compositions and medicaments containing such compounds, as well as methods for using such compounds in the treatment of conditions, diseases or disorders for which modulation of somatostatin activity would be effective.

Background of the Invention

Somatostatin is a peptide hormone that regulates the endocrine system and influences neurotransmission and cell proliferation by interacting with G-protein coupled somatostatin receptors and inhibiting the release of numerous secondary hormones. Six types of somatostatin receptor proteins have been identified (SSTR1, SSTR2a, SSTR2b, SSTR3, SSTR4, SSTR5) and are encoded by five different somatostatin receptor genes. Modulation of a particular somatostatin receptor subtype, or a combination thereof, holds promise for the treatment of conditions, diseases, or disorders that would benefit from modulation of somatostatin activity.

Brief summary of the present invention

In one aspect, the present invention describes a 3-[4-(4-aminopiperidin-1yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride compound, or a solvate thereof, of the following structure:

According to another aspect, the present invention describes crystalline 3-[4-(4aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or a solvate thereof. In some embodiments, crystalline 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or a solvate thereof is characterized by: X-ray powder diffraction (XRPD) with peaks at 4.5° 2-Theta, 9.1° 2-Theta, 10.2° 2-Theta, 16.3° 2-Theta, 18.4° 2-Theta and 19.1° 2-Theta ; X-ray powder diffraction (XRPD), essentially the same as shown in FIG. 1; a differential scanning calorimetry (DSC) thermogram with an endotherm starting at about 207° C. and peaking at about 220° C.; differential scanning calorimetry (DSC) thermogram essentially the same as shown in FIG. 2(a); thermogravimetric analysis (TGA) thermogram substantially the same as shown in FIG. 2(b); infrared (IR) with peaks at 2223 cm ^-1 , 1620 cm ^-1 , 1595 cm ^-1 , 1457 cm ^-1 , 1238 cm ^-1 , 1220 cm ^-1 and 1117 cm ^-1 ; infrared spectrum (IR), essentially the same as shown in Fig. 3; unchanged XRPD when heated to about 200°C, when exposed to more than 90% relative humidity for about 24 hours, or when exposed to about 75% RH and 40°C for one week, or a combination thereof; or their combinations.

In some embodiments, crystalline 3-[4-(4-aminopiperidin-1-yl)-3(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or solvate thereof is characterized by X-ray powder diffraction (XRPD) with peaks at 4.5° 2-Theta, 9.1° 2-Theta, 10.2° 2-Theta, 16.3° 2-Theta, 18.4° 2-Theta and 19.1° 2-Theta . In some embodiments, crystalline 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or a solvate thereof is characterized by an XRPD pattern that further includes peaks at 20.7° 2-Theta, 23.3° 2-Theta, 23.4° 2-Theta, 23.6° 2-Theta, 27.1° 2-Theta and 28.0° 2 -Theta. In some embodiments, crystalline 3-[4-(4-aminopiperidin-1-yl)-3-(3,5difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or solvate thereof is characterized by powder X-ray diffraction (XRPD), essentially the same as shown in Fig. 1. In some embodiments, crystalline 3-[4-(4-aminopiperidin-1-yl)-3-(3,5difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or a solvate thereof is characterized by a differential scanning calorimetry thermogram ( DSC) with an endotherm starting at about 207°C and peaking at about 220°C. In some embodiments, crystalline 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or solvate thereof is characterized by a differential scanning calorimetry thermogram ( DSC) is essentially the same as shown in FIG. 2(a). In some embodiments, crystalline 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2hydroxybenzonitrile monohydrochloride or a solvate thereof has an unchanged XRPD at heated to about 200°C. In some embodiments, crystalline 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or solvate thereof has unchanged XRPD when exposed to more than 90 % relative humidity for 24 hours and when exposed to about 75% RH and 40°C for one week. In some embodiments, crystalline 3-[4-(4-aminopiperidin-1-yl)-3-(3,5difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or a solvate thereof is characterized by an infrared spectrum (IR) with characteristic peaks at 2223 cm ^-1 , 1620 cm ^-1 , 1595 cm ^-1 , 1457 cm ^-1 , 1238 cm ^-1 , 1220 cm -1 and 1117 cm ^-1 . In some embodiments, crystalline 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or a solvate thereof is characterized by an infrared spectrum (IR) , essentially the same as IR. the spectrum shown in Fig. 3.

According to another aspect described in the present invention, there is provided a pharmaceutical composition comprising crystalline 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin6-yl]-2-hydroxybenzonitrile monohydrochloride or its solvate; and at least one pharmaceutically acceptable adjuvant. In some embodiments, the pharmaceutical composition is formulated for oral administration to a mammal. In some embodiments, the pharmaceutical composition is in the form of a solid pharmaceutical composition. In some embodiments, the pharmaceutical composition is in the form of a tablet, pill, or capsule.

According to another aspect described in the present invention, a method is provided for the preparation of crystalline 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or its solvate, which includes the steps in which:

(a) slurry 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile dihydrochloride in 5 volumes of isopropanol:water ( 1:1);

(i) heated the suspension (a) to about 45°C;

(ii) adding about 0.5 to about 1.2 equivalents of ammonium hydroxide solution, sodium bicarbonate solution, or sodium hydroxide solution to the heated slurry of step (a)(i) to give a pH of about 4.0-6.0;

(iii) water was added over about 2 hours to the mixture of step (a)(ii); and (iv) filtering the slurry of step (a)(iii) to give 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or its solvate; or (b) adding a suitable solvent to 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)quinolin-6-yl]-2-hydroxybenzonitrile;

(i) add about 1 equivalent of hydrochloric acid to a mixture of solvent and 3-[4-(4aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile (b); and (ii) filtering the solids from step (b)(ii) to give 3-[4-(4aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2 -hydroxybenzonitrile monohydrochloride or solvate thereof; or (c) stirring 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile dihydrochloride in from about 20 volumes to about 50 volumes water; and (i) filtering the solids of step (c) to give 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or a solvate thereof .

In some embodiments, an ammonium hydroxide solution is used in (a)(ii). In some embodiments, the amount of ammonium hydroxide solution used in (a)(ii) was about 0.8 equivalents and the pH reached was about 4.5-4.7.

In some embodiments, a suitable solvent in (b) is methanol, ethanol, isopropyl alcohol, acetone, methyl acetate, ethyl acetate, tetrahydrofuran, tetrahydropyran, water, or combinations thereof.

According to another aspect described in the present invention, a method for the synthesis of 3-[4(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile dihydrochloride is provided.

_NH22HCI

N which includes the processing step of compound A-VI

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NHBoc

Compound A-VI with hydrochloric acid in a suitable solvent.

According to another aspect described in the present invention, a method is provided for the synthesis of 3-[4(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride

which includes the steps in which (1) treated compound A-VI

NHBoc

Compound A-VI with hydrochloric acid in a suitable solvent to give 3-[4-(4-aminopiperidin1-yl)-3-(3,5-difluorophenyl)quinolin-6-yl]-2-hydroxybenzonitrile dihydrochloride; and (2) treating 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile dihydrochloride with aqueous ammonia to give 3-[4- (4-aminopiperidin-1-yl)-3-(3,5difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride.

In some embodiments, a suitable solvent is isopropyl alcohol, ethyl acetate, or isopropyl acetate. In some embodiments, a suitable solvent is isopropyl alcohol.

According to another aspect described in the present invention provides a method for obtaining compounds A-VI

NHBoc

The compound A-VI which includes the steps in which (1) the reaction of the compound A-IV NHBoc was carried out

Compound A-IV with Compound 1

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HE

Compound 1 where

B is boronic acid, boronic acid ester or trifluoroborate;

in the presence of an addition catalyst, a suitable base, and in a suitable solvent to give compound A-V

NHBoc

A-V connection and

(2) reacting compound A-V with 3,5-difluorophenylboronic acid

B(OH) ₂

in the presence of an addition catalyst, a suitable base and in a suitable solvent to give compound A-VI.

In some embodiments, B is boronic acid or trifluoroborate.

In some embodiments, B is boronic acid. In some embodiments, B is a trifluoroborate.

In some embodiments, the addition catalyst of step (1) is a palladium catalyst; a suitable base of step (1) is triethylamine, diisopropylethylamine, 1,2,2,6,6-pentamethylpiperidine, tributylamine, sodium bicarbonate, Na ₂ CO ₃ , K ₂ CO ₃ , Cs ₂ CO ₃ , NaOAc, KOAc, Ba( OH) ₂ , Na ₃ PO4 or K3PO4; and a suitable solvent of step (1) is acetonitrile, dimethylformamide, ethanol, tetrahydrofuran, isopropyl alcohol, 1,4-dioxane, water, or combinations thereof. In some embodiments, step (1) was performed at a temperature of about 80°C. In some embodiments, step (1) is performed at a temperature of about 80-85°C.

In some embodiments, the addition catalyst of step (1) is a palladium catalyst; a suitable base of step (1) is K ₂ CO ₃ ; and a suitable solvent of step (1) is a mixture of 1,4-dioxane and water.

In some embodiments, the addition catalyst of step (2) is a palladium catalyst; a suitable base of step (2) is triethylamine, diisopropylethylamine, 1,2,2,6,6-pentamethylpiperidine, tributylamine, sodium bicarbonate, Na ₂ CO ₃ , K ₂ CO ₃ , Cs ₂ CO ₃ , NaOAc, KOAc, Ba( OH) ₂ , Na ₃ PO ₄ or K ₃ PO ₄ ; and a suitable solvent of step (2) is acetonitrile, dimethylformamide, ethanol, tetrahydrofuran, isopropyl alcohol, 1,4-dioxane, water, or combinations thereof. In some embodiments, step (2) is performed at a temperature of about 90°C to about 100°C. In some embodiments, the addition catalyst of step (2) is a palladium catalyst; a suitable base of step (2) is K ₂ CO ₃ ; and a suitable solvent of step (2) is a mixture of 1,4dioxane and water.

In some embodiments, Compound A-V is isolated prior to step (2).

In some embodiments, Compound A-V was not isolated prior to step (2).

In some embodiments, the method further comprises recrystallizing Compound A-VI from a suitable solvent. In some embodiments, a suitable solvent is methyl acetate, ethyl acetate, isopropyl acetate, methanol, ethanol, isopropyl alcohol, dichloromethane/petroleum ether, acetonitrile, tetrahydrofuran/water, tetrahydrofuran/petroleum ether, dimethylformamide/water, dichloromethane/methyl tert-butyl ether, methanol/ methyl tert-butyl ether, methyl tert-butyl ether or toluene. In some embodiments, a suitable solvent is ethyl acetate or isopropyl acetate.

In some embodiments, the method further comprises recrystallizing Compound A-VI with a metal scavenger. In some embodiments, the metal scavenger comprises SiO ₂ , carbon, L-cysteine aqueous solution, Silicycle metal scavenger,

Si-thiol, SiliaBond DMT or SiliaBond cysteine.

According to another aspect described in the present invention provides a method for obtaining compounds A-IV

NHBoc which includes steps in which (1) Compound A-I was chlorinated

ABOUT

H

Compound A-I with a suitable chlorinating agent in a suitable solvent to obtain compound A-II

HE

Connection A-P;

(2) brominate compound A-II with a suitable brominating agent in a suitable solvent to give compound A-III

Vg

Α-ΠΙ connection;

and (3) combining 4-(N-Boc amino)piperidine with Compound A-III in the presence of a suitable base and in a suitable solvent to give Compound A-IV; or (i) combine 4-(N-Boc amino)piperidine with 6-bromo-4-chloroquinoline in the presence of a suitable base and in a suitable solvent to give compound 4

NHBoc

Compound 4;

and (ii) chlorinating compound 4 with a suitable chlorinating agent in a suitable solvent to give compound A-IV.

In some embodiments, the chlorinating agent of step (1) is N-chlorosuccinimide, trichloroisocyanuric acid, sulfuryl chloride, chlorine, sodium hypochlorite, calcium hypochlorite, hypochlorous acid, or 2,3,4,5,6,6-hexachloro-2,4- cyclohexadien-1-one; and a suitable solvent of step (1) is acetic acid, water, ethanol, methanol, toluene, dichloromethane, tetrahydrofuran, dioxane, or N,N-dimethylformamide.

In some embodiments, the chlorinating agent of step (1) is N-chlorosuccinimide; and a suitable solvent of step (1) is acetic acid.

In some embodiments, the brominating agent of step (2) is phosphorus tribromide, phosphorus oxybromide, hydrobromic acid, bromine, or dibromotriphenyl phosphorane; and a suitable solvent of step (2) is acetonitrile, water, ethanol, isopropanol, dichloromethane, toluene, N,N-dimethylformamide, acetic acid or acetone.

In some embodiments, the brominating agent of step (2) is phosphorus tribromide; and a suitable solvent of step (2) is N,N-dimethylformamide.

In some embodiments, a suitable base of step (3) is triethylamine, diisopropylethylamine, 1,8-diazabicycloundec-7-ene, 1,2,2,6,6-pentamethylpiperidine, tributylamine, sodium bicarbonate, Na ₂ CO ₃ , K ₂ CO ₃ or Cs ₂ CO ₃ ; and a suitable solvent of step (3) is N,N-dimethylformamide, N,N-dimethylacetamide, dimethylsulfoxide, dichloromethane, chloroform, carbon tetrachloride, dioxane, tetrahydrofuran, toluene, acetonitrile, ethanol or isopropanol.

In some embodiments, the base of step (3) is diisopropylethylamine; and a suitable solvent of step (3) is dimethyl sulfoxide.

In some embodiments, a suitable base of step (i) is triethylamine, diisopropylethylamine, 1,8-diazabicycloundec-7-ene, 1,2,2,6,6-pentamethylpiperidine, tributylamine, sodium bicarbonate, Na ₂ CO ₃ , K2CO3 or _{Cs2CO3 ;} and a suitable solvent of step (i) is N,N-dimethylformamide, N,N-dimethylacetamide, dimethylsulfoxide, dichloromethane, chloroform, carbon tetrachloride, dioxane, tetrahydrofuran, toluene, acetonitrile, ethanol or isopropanol.

In some embodiments, the base of step (i) is K2CO3; and a suitable solvent of step (i) is N,N-dimethylformamide.

In some embodiments, the chlorinating agent of step (ii) is N-chlorosuccinimide, trichloroisocyanuric acid, sulfuryl chloride, chlorine, sodium hypochlorite, calcium hypochlorite, hypochlorous acid, or 2,3,4,5,6,6-hexachloro-2,4- cyclohexadien-1-one; and a suitable solvent of step (ii) is acetic acid, water, ethanol, methanol, toluene, dichloromethane, tetrahydrofuran, dioxane or N,N-dimethylformamide.

In some embodiments, the chlorinating agent of step (ii) is N-chlorosuccinimide; and a suitable solvent of step (ii) is toluene.

According to another aspect described in the present invention, a method for obtaining compound 1 is provided.

HE

B. /CN

Compound 1 where B is a boronic acid or an ester of boronic acid, which includes steps in which (1) the hydroxyl group of compound 2 is protected

HE

Br\ /CN

Compound 2;

with a suitable protecting group (PG) to give compound 2a

OPG'

Br\ /CN

Compound 2a;

(2) reacting compound 2a with a borylating agent under suitable reaction conditions; and (3) removing the protecting group (PG') to give Compound 1.

In some embodiments, B is boronic acid.

In some embodiments, the method further comprises converting B to a trifluoroborate.

In some embodiments, the boryling agent is triisopropylborate, trimethylborate, tetrahydroxydiborone, pinacolborane, catecholborane, bis(neopentylglycolate)diborone, bis(pinacolato)diborone, bis(hexyleneglycolate)diborone, bis(catecholate)diborone, 4,4,5,5- tetramethyl-1,3,2-dioxaborolane, 4,6,6-trimethyl-1,3,2-dioxaborinan, diisopropylamine borane, bis(neopentylglycolate)diborone, bis(catecholato)diborone or bis(pinacolato)diborone.

In some embodiments, suitable reaction conditions (2) include the use of halogen-metal exchange reagents. In some embodiments, suitable reaction conditions (2) include the use of halogen-metal exchange reagents selected from Grignard reagents and alkyl-lithium reagents. In some embodiments, suitable reaction conditions (2) include the use of isopropyl magnesium chloride in tetrahydrofuran.

In some embodiments, the boryling agent is triisopropyl borate and suitable reaction conditions (2) include the use of isopropyl magnesium chloride in tetrahydrofuran.

In some embodiments, suitable reaction conditions (2) include the use of transition metal mediated reaction conditions.

In some embodiments, suitable reaction conditions (2) include the use of palladium metal mediated reaction conditions.

In some embodiments, a suitable protecting group (PG') is methoxymethyl, ethoxyethyl, methoxypropyl, benzyloxymethyl, 2-methoxyethoxymethyl, benzyl, p-methoxybenzyl, 2-naphthylmethyl, methyl, allyl, tetrahydropyranyl, acetyl, benzoyl, 2,2,2 -trichloroethyl carbonyl, trimethylsilyl, triethylsilyl, triisopropylsilyl, t-butyldimethylsilyl or t-butyldiphenylsilyl.

In some embodiments, a suitable protecting group (PG') is methoxymethyl, 2-methoxyethoxymethyl, benzyl, p-methoxybenzyl, methyl, allyl, tetrahydropyran-2-yl, [2-(trimethylsilyl)ethoxy]methyl, trimethylsilyl, triethylsilyl, triisopropylsilyl , tert-butyldimethylsilyl or tert-butyldiphenylsilyl.

In some embodiments, a suitable protecting group (PG') is methoxymethyl, ethoxyethyl, methoxypropyl, benzyloxymethyl, 2-methoxyethoxymethyl, benzyl, p-methoxybenzyl, 2-naphthylmethyl, methyl, allyl, or tetrahydropyranyl. In some embodiments, a suitable protecting group (PG') is methoxymethyl, ethoxyethyl, or 2methoxyethoxymethyl. In some embodiments, a suitable protecting group (PG') is methoxymethyl.

In some embodiments, a suitable protecting group (PG') is acetyl, benzoyl, or 2,2,2-trichloroethylcarbonyl. In some embodiments, a suitable protecting group (PG) is acetyl.

In some embodiments, deprotection in step (3) was performed by treatment with hydrochloric acid, hydrobromic acid, acetic acid, trifluoroacetic acid, p-toluenesulfonic acid, ZnBr2, hydrogen over Pd/C, 2,3-dichloro-5,6dicyano-1 ,4-benzoquinone (DDQ), boron tribromide, boron trichloride, trimethylsilyl iodide, Pd(PPh ₃ ) ₄ , tetra-n-butylammonium fluoride (TBAF) or HF-pyridine.

In some embodiments, a suitable protecting group (PG') is methoxymethyl; and the removal of the protecting group in step (3) was performed by treatment with hydrochloric acid.

Articles are provided that include a packaging material, a somatostatin modulator or a pharmaceutically acceptable salt or solvate thereof as described in the present invention, in the packaging material and a label indicating that the somatostatin modulator or a pharmaceutically acceptable salt or solvate thereof is used to modulate one or more somatostatin receptor protein subtypes, or to treat, prevent or alleviate one or more symptoms of a disease or condition for which modulation of one or more somatostatin receptor protein subtypes would be effective.

Other objects, features, and advantages of the compounds, methods, and compositions described herein will become apparent from the following detailed description. It should be appreciated, however, that the detailed description and specific examples, while describing specific embodiments, are for illustrative purposes only, as various changes and modifications to the spirit and scope of the present disclosure will become apparent to those skilled in the art from this detailed description.

Brief description of the graphic material

Fig. 1. Powder X-ray diffraction (XRPD) compound A, mono-HCl salt.

Fig. 2(a). Differential Scanning Calorimetry (DSC) thermogram.

Fig. 2(b). Thermogravimetric analysis/differential scanning calorimetry (TGA/DSC) thermogram of compound A, mono-HCl salt.

Fig. 3. Infrared spectrum (IR) of compound A, mono-HCl salt.

Fig. 4. Powder X-ray diffraction (XRPD) compound A, di-HCl salt.

Fig. 5(a). Differential scanning calorimetry (DSC) thermogram of compound A, diHCl salt.

Fig. 5(b). Thermogram of thermogravimetric analysis (TGA) of compound A, di-HCl salt.

Fig. 6. Infrared spectrum (IR) of compound A, di-HCl salt.

Fig. 7(a). X-ray powder diffraction pattern (XRPD) A of compound A, free base.

Fig. 7(b). X-ray powder diffraction pattern (XRPD) B compound A, free

- 7 042196 base.

Fig. 7(c). X-ray powder diffraction (XRPD) Compound A, free base.

Fig. 8(a). Differential scanning calorimetry (DSC) thermogram of sample C of compound A, free base.

Fig. 8(b). Thermogram of thermogravimetric analysis (TGA) of sample C of compound A, free base.

Fig. 9. Powder X-ray diffraction (XRPD) of compound A, mono-HCl salt before (lower spectra) and after (upper spectra) a dynamic vapor sorption (DVS) study from 2 to 95% relative humidity (RH).

Fig. 10. Powder X-ray diffraction (XRPD) of compound A, di-HCl salt before (lower spectra) and after (upper spectra) a dynamic vapor sorption (DVS) study from 2 to 95% relative humidity (RH).

Detailed disclosure of the present invention

Somatostatin (SST), also known as somatotropin release inhibitory factor (SRIF), was originally isolated as a 14 amino acid peptide from the sheep hypothalamus (Brazeau et al., Science 179, 77-79, 1973). Subsequently, a 28 amino acid N-terminally extended peptide with biological activity similar to 14 amino acid somatostatin was isolated (Pradayrol et al., FEBS Letters, 109, 55-58, 1980; Esch et al., Proc. Natl. Acad. Sci USA, 77, 6827-6831, 1980). SST is a regulatory peptide produced by several cell types in response to other neuropeptides, neurotransmitters, hormones, cytokines, and growth factors. SST acts through both endocrine and paracrine pathways with effects on its target cells. Many of these effects are associated with inhibition of the secretion of other hormones, predominantly growth hormone (GH). They are produced by a wide variety of cell types in the central nervous system (CNS) and intestines and have numerous functions, including modulating the secretion of growth hormone (GH), insulin, glucagon, and many other hormones that are antiproliferative.

These pleiotropic actions of somatostatins are mediated by six somatostatin receptor proteins (SSTR1, SSTR2a, SSTR2b, SSTR3, SSTR4, SSTR5). The six somatostatin receptor proteins are encoded by five different somatostatin receptor genes (Reisine and Bell, Endocr Rev. 16, 427-442, 1995; Patel and Srikant, Trends Endocrinol Metab 8, 398-405, 1997). All receptors are members of the class A subgroup GPCR superfamily. The SST2A receptor is the most widely expressed subtype in human tumors and is the dominant receptor by which GH secretion is suppressed. Unless otherwise indicated, the term SSTR2 means SSTR2a.

It is possible to selectively modulate any of the somatostatin receptor subtypes or a combination thereof. In some embodiments, selective modulation of any of the somatostatin receptor subtypes with respect to other somatostatin receptor subtypes, or a combination thereof, is useful in a number of clinical applications. In some embodiments, selective modulation of any of the somatostatin receptor subtypes relative to other somatostatin receptor subtypes reduces unwanted side effects in a number of clinical applications.

For example, modulation of SSTR2 activity mediates inhibition of growth hormone (GH) release from the anterior pituitary and glucagon release from the pancreas. SSTR2 is also involved in many other biological functions such as, but not limited to, cell proliferation, nociception, inflammation, and angiogenesis. In some embodiments, the SSTR2 selective modulator is used in the treatment of acromegaly, intestinal neuroendocrine tumors, pain, neuropathies, nephropathies and inflammation, and retinopathy resulting from abnormal blood vessel growth. In some embodiments, the SSTR2 selective modulator is used in the treatment of arthritis, pain, cancer, inflammatory bowel disease, irritable bowel syndrome, Crohn's disease, Cushing's disease, acute lung injury, acute respiratory distress syndrome, and eye disorders such as age-related macular degeneration (AMD). ), diabetic retinopathy, diabetic macular edema, and Graves' ophthalmopathy, among others.

In some embodiments, SSTR4 agonists have anti-inflammatory and anti-nociceptive effects.

In some embodiments, SSTR3 agonists inhibit insulin secretion.

In some embodiments, SSTR5 agonists inhibit insulin secretion. In addition, SSTR5 was also involved in the modulation of growth hormone release.

The peptide somatostatin and its receptor subtypes are also widely expressed in the brain, and disruption or reduction of its activity is potentially associated with several psychiatric and neurodegenerative diseases. For example, somatostatin concentrations in the cerebral cortex and hippocampus are reduced in schizophrenics, and one of the most consistent neuropathological findings in this group of patients is a deficiency of somatostatin-expressing cortical inhibitory interneurons. Somatostatin is also highly expressed in areas of the brain associated with seizures and plays an important role in epilepsy. Somatostatin levels are reduced in the hippocampus of Alzheimer's and Parkinson's patients, suggesting restoration of somatostatin signaling as a potential drug target in neurodegeneration.

In one aspect, the compounds described herein are SSTR2 modulators. In some embodiments, the compounds described herein selectively modulate SSTR2 activity for other somatostatin receptors.

In some embodiments, the compounds described herein are for oral administration to a mammal in need of somatostatin modulator treatment.

In some embodiments, the somatostatin receptor modulators described herein are useful in a wide range of therapeutic applications. In some embodiments, the somatostatin receptor modulators described herein are used in the treatment of a number of diseases or conditions such as, but not limited to, acromegaly, neuroendocrine tumors, retinopathy and other ocular disorders, neuropathy, nephropathy, respiratory diseases, cancers, pain, neurodegenerative diseases, inflammatory diseases, and psychiatric and neurodegenerative disorders. In some embodiments, the somatostatin receptor modulators described herein are used in the treatment of acromegaly in a mammal.

In some embodiments, the somatostatin receptor modulators described herein inhibit the secretion of various hormones and trophic factors in mammals. In some embodiments, the compounds are used to suppress certain endocrine secretions such as, but not limited to, GH, insulin, glucagon, and prolactin. Suppression of certain endocrine secretions is useful in the treatment of disorders such as acromegaly; endocrine tumors and carcinoids, VIPomas, insulinomas and glucagonomas; or diabetes mellitus and diabetes-related pathologies, including retinopathy, neuropathy and nephropathy. In some embodiments, the somatostatin receptor modulators described herein are used to suppress secretions in the pancreas, stomach, and intestines in the treatment of disorders such as pancreatitis, fistulas, bleeding ulcers, and diarrhea associated with diseases such as AIDS or cholera. Disorders involving autocrine or paracrine secretions of trophic factors such as IGF-1 (as well as certain endocrine factors) that can be treated by administering the compounds described in the present invention, the compounds include breast, prostate and lung cancers (both small cell, and non-small cell epidermoids), as well as hepatomas, neuroblastomas, adenocarcinomas of the large intestine and pancreas (ductal type), chondrosarcomas and melanomas, diabetic retinopathy and atherosclerosis associated with vascular implants and restenosis after angioplasty.

In some embodiments, the somatostatin receptor modulators described herein are used to suppress neurogenic inflammatory mediators (eg, substance P or tachykinins) and may be used in the treatment of rheumatoid arthritis; psoriasis; local inflammation, for example, associated with sunburn, eczema or other sources of itching; inflammatory bowel disease; irritable bowel syndrome; allergies, including asthma and other respiratory diseases. In some embodiments, the somatostatin receptor modulators described herein function as neuromodulators in the central nervous system and are useful in the treatment of Alzheimer's disease and other forms of dementia, pain and headaches. In some embodiments, the somatostatin receptor modulators described herein provide cytoprotection for disorders involving visceral blood flow, including cirrhosis and esophageal varices.

Compound A is a somatostatin modulator that is useful in the methods of treatment described in the present invention.

Connection A.

Compound A as used herein refers to 3-(4-(4-aminopiperidin-1-yl)-3(3,5-difluorophenyl)-quinolin-6-yl)-2-hydroxybenzonitrile which has the chemical structure shown below .

- 9 042196

Compound A

In some embodiments, Compound A is amorphous.

In some embodiments, Compound A is crystalline.

In some embodiments, compound A is crystalline and has X-ray powder diffraction of the sample with peaks at 9.2° 2-Theta, 12.3° 2-Theta, 14.4° 2-Theta, and 24.0° 2-Theta. In some embodiments, Compound A is crystalline and has an X-ray powder diffraction pattern substantially similar to the XRPD shown in FIG. 7(a).

In some embodiments, compound A is crystalline and has X-ray powder diffraction of the sample with peaks at 5.9° 2-Theta, 13.9° 2-Theta, 14.2° 2Theta, 17.5° 2-Theta, and 24.6 ° 2-Theta. In some embodiments, Compound A is crystalline and has an X-ray powder diffraction pattern substantially similar to the XRPD shown in FIG. 7(b).

In some embodiments, Compound A is crystalline and is characterized as containing: X-ray powder diffraction (XRPD) with peaks at 7.2° 2Theta, 8.3° 2-Theta, 10.9° 2-Theta, and 12.0° 2- Theta; powder x-ray diffraction (XRPD) substantially the same as shown in FIG. 7(c); a differential scanning calorimetry (DSC) thermogram with an endotherm starting at about 128° C. and peaking at about 145° C.; differential scanning calorimetry (DSC) thermogram substantially the same as shown in FIG. 8(a); differential scanning calorimetry (DSC) thermogram substantially the same as shown in FIG. 8(a); a thermogravimetric analysis (TGA) thermogram substantially the same as shown in FIG. 8(b); or their combinations.

In some embodiments, Compound A is crystalline and is characterized as having at least one of the following properties:

a) X-ray powder diffraction pattern (XRPD) with peaks at 7.2° 2-Theta, 8.3° 2-Theta, 10.9° 2-Theta and 12.0° 2-Theta;

b) X-ray powder diffraction (XRPD), essentially as shown in FIG. 7(c);

c) differential scanning calorimetry (DSC) thermogram with endotherm starting at about 128°C and peaking at about 145°C;

d) a differential scanning calorimetry (DSC) thermogram essentially as shown in FIG. 8(a).

In some embodiments, Compound A is crystalline and is characterized as having at least two properties selected from a)-d). In some embodiments, Compound A is crystalline and is characterized as having at least three properties selected from a)-d). In some embodiments, Compound A is crystalline and is characterized as having properties a), b), c) and d).

In some embodiments, compound A is crystalline and is characterized by X-ray powder diffraction (XRPD) with peaks at 7.2° 2-Theta, 8.3° 2-Theta, 10.9° 2-Theta, and 12.0° 2-Theta . In some embodiments, compound A is crystalline and has an X-ray powder diffraction (XRPD) pattern substantially the same as shown in FIG. 7(c). In some embodiments, compound A is crystalline and is characterized by a differential scanning calorimetry (DSC) thermogram with an endotherm starting at about 128°C and peaking at about 145°C. In some embodiments, compound A is crystalline and has a differential scanning calorimetry (DSC) thermogram substantially the same as shown in FIG. 8(a).

In some embodiments, the present invention provides a pharmaceutically acceptable salt of Compound A. In some embodiments, the pharmaceutically acceptable salt of Compound A is the monohydrochloride salt (compound A-HCl):

- 10 042196

Compound A-HC1

In some embodiments, the A-HCl compound is amorphous.

In some embodiments, the A-HCl compound is crystalline.

In some embodiments, compound A-HCl is crystalline and is characterized as having: X-ray powder diffraction (XRPD) peaks at 4.5° 2-Theta, 9.1° 2-Theta, 10.2° 2-Theta, 16, 3° 2-Theta, 18.4° 2-Theta and 19.1° 2-Theta; X-ray powder diffraction (XRPD) is essentially the same as shown in FIG. 1; a differential scanning calorimetry (DSC) thermogram with an endotherm starting at about 207° C. and peaking at about 220° C.; differential scanning calorimetry (DSC) thermogram is essentially the same as shown in FIG. 2(a); thermogravimetric analysis (TGA) thermogram is essentially the same as shown in FIG. 2(b); infrared (IR) with peaks at 2223 cm ^-1 , 1620 cm ^-1 , 1595 cm ^-1 , 1457 cm ^-1 , 1238 cm ^-1 , 1220 cm ^-1 and 1117 cm ^-1 ; infrared spectrum (IR) is essentially the same as shown in FIG. 3; unchanged XRPD when heated to about 200°C, when exposed to more than 90% relative humidity for about 24 hours, or when exposed to about 75% RH and 40°C for one week, or a combination thereof; or their combinations.

In some embodiments, the A-HCl compound is crystalline and is characterized as having at least one of the following properties:

a) X-ray powder diffraction pattern (XRPD) peaking at 4.5° 2-Theta, 9.1° 2-Theta, 10.2° 2-Theta, 16.3° 2-Theta, 18.4° 2-Theta and 19.1° 2-Theta;

b) X-ray powder diffraction (XRPD) is essentially the same as shown in FIG. 1;

c) differential scanning calorimetry (DSC) thermogram with an endotherm starting at about 207° C. and peaking at about 220° C.;

d) differential scanning calorimetry (DSC) thermogram essentially the same as shown in FIG. 2(a);

e) infrared spectrum (IR) with characteristic peaks at 2223 cm ^-1 , 1620 cm ^-1 , 1595 cm ^-1 , 1457 cm ^-1 , 1238 cm ^-1 , 1220 cm ^-1 and 1117 cm ^-1 ;

f) infrared spectrum (IR) is essentially the same as shown in FIG. 3;

g) reversible water absorption (-4.5% w/w) from 2 to 95% relative humidity (RH);

h) reversible water absorption (~2.3%) from 15 to 75% relative humidity (RH);

i) unchanged XRPD after DVS analysis at 90% RH and room temperature for 24 hours; or j) unchanged XRPD after DVS analysis at 75% RH and 40°C for one week.

In some embodiments, compound A-HCl is crystalline and is characterized as having at least two properties selected from a)-j). In some embodiments, compound A-HCl is crystalline and is characterized as having at least three properties selected from a)-j). In some embodiments, compound A-HCl is crystalline and is characterized as having at least four properties selected from a)-j). In some embodiments, compound A-HCl is crystalline and is characterized as having at least five properties selected from a)-j). In some embodiments, the AHCl compound is crystalline and is characterized as having at least six properties selected from a)-j). In some embodiments, compound A-HCl is crystalline and is characterized as having at least seven properties selected from a)-j). In some embodiments, compound A-HCl is crystalline and is characterized as having at least eight properties selected from a)-j). In some embodiments, compound A-HCl is crystalline and is characterized as having at least nine properties selected from a)-j). In some embodiments, the A-HCl compound is crystalline and is characterized as having properties a), b), c), d), e), f), g), h), i) and j).

In some embodiments, compound A-HCl is crystalline and is characterized by X-ray powder diffraction (XRPD) with peaks at 4.5° 2-Theta, 9.1° 2Theta, 10.2° 2-Theta, 16.3° 2-Theta , 18.4° 2-Theta and 19.1° 2-Theta. In some embodiments, compound A-HCl is crystalline and has an X-ray powder diffraction (XRPD) pattern substantially the same as shown in FIG. 1. In some embodiments, the A-HCl compound is crystalline and has a diff thermogram

- 11 042196 Rational Scanning Calorimetry (DSC) with an endotherm starting at about 207°C and peaking at about 220°C. In some embodiments, compound A-HCl is crystalline and has a differential scanning calorimetry (DSC) thermogram substantially the same as shown in FIG. 2(a). In some embodiments, the A-HCl compound is crystalline and is characterized by an infrared spectrum (IR) with characteristic peaks at 2223 cm ^-1 , 1620 cm ^-1 , 1595 cm ^-1 , 1457 cm ^-1 , 1238 cm ^-1 , 1220 cm ^-1 and 1117 cm ^-1 . In some embodiments, compound A-HCl is crystalline and has an infrared (IR) spectrum substantially the same as shown in FIG. 3. In some embodiments, compound A-HCl is crystalline and has reversible water uptake (~4.5% w/w) from 2 to 95% relative humidity (RH). In some embodiments, the A-HCl compound is crystalline and has a reversible water uptake (~2.3%) from 15 to 75% relative humidity (RH). In some embodiments, compound A-HCl is crystalline and has unchanged XRPD after DVS analysis at 90% RH and room temperature for 24 hours. In some embodiments, compound A-HCl is crystalline and has unchanged XRPD after DVS analysis at 75% RH and 40°C for one week.

In some embodiments, the present invention provides a pharmaceutically acceptable salt of Compound A. In some embodiments, the pharmaceutically acceptable salt of Compound A is the dihydrochloride salt (Compound A-2HC1)

Compound A-2HC1

In some embodiments, the A-2HC1 compound is amorphous.

In some embodiments, the A-2HC1 compound is crystalline.

In some embodiments, compound A-2HC1 is crystalline and is characterized as having: X-ray powder diffraction (XRPD) peaks at 5.4° 2-Theta and 7.3° 2-Theta; X-ray powder diffraction (XRPD) is essentially the same as shown in FIG. 4; a differential scanning calorimetry (DSC) thermogram with an endotherm starting at about 233° C. and peaking at about 252° C.; differential scanning calorimetry (DSC) thermogram is essentially the same as shown in FIG. 5(a); thermogravimetric analysis (TGA) thermogram is essentially the same as shown in FIG. 5(b); infrared spectrum (IR) with characteristic peaks at 2227 cm ^-1 , 1620 cm ^-1 , 1594 cm ^-1 , 1456 cm ^-1 , 1439 cm ^-1 , 1321 cm ^-1 and 1122 cm ^-1 ; infrared spectrum (IR) is essentially the same as shown in FIG. 6; reversible water absorption (~18% w/w) from 2 to 95% relative humidity (RH); reversible water absorption (~9% w/w) from 2 to 95% relative humidity (RH); unchanged XRPD after DVS analysis at 90% RH and room temperature for 24 hours; unchanged XRPD after DVS analysis at 75% RH and 40°C for one week; or their combinations.

In some embodiments, the A-2HC1 compound is crystalline and has at least one of the following properties:

a) X-ray powder diffraction pattern (XRPD) with peaks at 5.4° 2-Theta and 7.3° 2-Theta;

b) X-ray powder diffraction (XRPD) is essentially the same as shown in FIG. 4;

c) differential scanning calorimetry (DSC) thermogram with an endotherm starting at about 233°C and peaking at about 252°C;

d) differential scanning calorimetry (DSC) thermogram essentially the same as shown in FIG. 5(a);

e) infrared spectrum (IR) with characteristic peaks at 2227 cm ^-1 , 1620 cm ^-1 , 1594 cm ^-1 , 1456 cm ^-1 , 1439 cm ^-1 , 1321 cm ^-1 and 1122 cm ^-1 ;

f) infrared spectrum (IR) is essentially the same as shown in FIG. 6;

g) reversible water absorption (~18% w/w) from 2 to 95% relative humidity (RH);

h) reversible water absorption (~9% w/w) from 2 to 95% relative humidity (RH);

i) unchanged XRPD after DVS analysis at 90% RH and room temperature for 24 hours; or j) unchanged XRPD after DVS analysis at 75% RH and 40°C for one week.

In some embodiments, the A-2HC1 compound is crystalline and is characterized as having at least two properties selected from a)-j). In some embodiments, the A-2HC1 compound is crystalline and is characterized as having at least three properties selected from a)-j). In some embodiments, the A-2HC1 compound is crystalline and is characterized as having

- 12 042196 at least four properties selected from a)-j). In some embodiments, the A-2HC1 compound is crystalline and is characterized as having at least five properties selected from a)-j). In some embodiments, the A2HCl compound is crystalline and is characterized as having at least six properties selected from a)-j). In some embodiments, compound A-2HCl is crystalline and is characterized as having at least seven properties selected from a)-j). In some embodiments, compound A-2HCl is crystalline and is characterized as having at least eight properties selected from a)-j). In some embodiments, compound A-2HCl is crystalline and is characterized as having at least nine properties selected from a)-j). In some embodiments, compound A-2HCl is crystalline and is characterized as having properties a), b), c), d), e), f), g), h), i) and j).

In some embodiments, compound A-2HCl is crystalline and is characterized by powder X-ray diffraction (XRPD) with characteristic peaks at 5.4° 2-Theta and 7.3° 2-Theta. In some embodiments, compound A-2HCl is crystalline and has an X-ray powder diffraction (XRPD) pattern substantially the same as shown in FIG. 4. In some embodiments, the A2HCl compound is crystalline and is characterized by a differential scanning calorimetry (DSC) thermogram with an endotherm starting at about 233°C and peaking at about 252°C. In some embodiments, compound A-2HCl is crystalline and has a differential scanning calorimetry (DSC) thermogram substantially the same as shown in FIG. 5(a). In some embodiments, compound A-2HCl is crystalline and has an infrared (IR) spectrum with characteristic peaks at 2227 cm ^-1 , 1620 cm ^-1 , 1594 cm ^-1 , 1456 cm ^-1 , 1439 cm ^-1 , 1321 cm ^-1 and 1122 cm ^-1 . In some embodiments, compound A-2HCl is crystalline and has an infrared (IR) spectrum substantially the same as shown in FIG. 6. In some embodiments, compound A-2HCl is crystalline and has reversible water uptake (~18% w/w) from 2 to 95% relative humidity (RH). In some embodiments, compound A-2HCl is crystalline and has reversible water uptake (~9% w/w) from 2 to 95% relative humidity (RH). In some embodiments, compound A-2HCl is crystalline and has unchanged XRPD after DVS analysis at 90% RH and room temperature for 24 hours. In some embodiments, compound A-2HCl is crystalline and has XRPD after DVS analysis at 75% RH and 40°C for one week.

Synthesis of compound A, compound A-HCl and compound A-2HCl.

The compounds described in the present invention were synthesized using standard synthetic procedures or using methods known in the art in combination with the methods described in the present invention. Unless otherwise noted, traditional methods of mass spectroscopy, NMR, HPLC were used.

Compounds were prepared using standard organic chemistry techniques such as those described, for example, in March's Advanced Organic Chemistry, ^6th Edition, John Wiley and Sons, Inc. Alternative reaction conditions for synthesis transformations described herein can be used, such as solvent variations, reaction temperature, reaction time, as well as various chemicals and other reaction conditions.

In the reactions described, it may be necessary to protect reactive functional groups, for example hydroxy or amino groups, where they are needed in the final product, to avoid their undesirable participation in the reactions. Detailed descriptions of techniques applicable to the formation of protecting groups and their removal are described in Greene and Wuts, Protective Groups in Organic Synthesis, ^3rd Ed., John Wiley & Sons, New York, NY, 1999, and Kocienski, Protective Groups, Thieme Verlag , New York, NY, 1994, which are incorporated herein by reference.

The present invention describes methods for the synthesis of Compound A, Compound A-HCl, and Compound A-2HCl as outlined in Scheme A.

- 13 042196

Scheme A

In some embodiments, chlorination of compound AI results in compound A-II. Compound A-II was reacted with a brominating or chlorinating agent to give compound A-IIIa (where X is Cl or Br). In some embodiments, Compound A-II is reacted with a brominating agent to produce Compound A-IIIa, where X is Br (ie, Compound A-III). Coupling compound A-IIIa with 4-(N-PG-amino)piperidine gave compound A-IVa (where PG is the appropriate amino protecting group). Compound A-IVa was subjected to a Suzuki reaction with compound 1 to give compound A-Va, which was subjected to a second Suzuki reaction with (3,5-difluorophenyl)B (where each B is independently a boronic acid, boronate ester, or trifluoroborate) to give the compound A-VIa. In some embodiments, each B is boronic acid. In some embodiments, (3,5-difluorophenyl)B is (3,5-difluorophenyl)boronic acid. In some embodiments, the same Pd catalyst was used in both Suzuki reactions. In other embodiments, different Pd catalysts were used in each Suzuki reaction. In some embodiments, each Suzuki reaction was carried out in the same solvent. In other embodiments, each Suzuki reaction was run in a different solvent. In some embodiments, Compound A-Va is isolated prior to the second Suzuki reaction. In other embodiments, two Suzuki reactions were performed in the same reaction vessel without isolating or purifying compound A-Va. In some embodiments, residual palladium was removed from Compound A-VIa using a palladium scavenger such as SiO ₂ , carbon, L-cysteine, SilicaBond cysteine, Si-thiol, SilicaBond DMT, or the like.

Deprotection of the PG group of compound A-VIa and treatment with HCl gave compound A-2HC1, which was converted to compound A-HCl. In some embodiments, Compound A-HCl or Compound A-2HC1 is treated with an appropriate base, such as sodium hydroxide, sodium carbonate, sodium bicarbonate, or the like, to provide Compound A (free base form).

In some embodiments, if PG is t-butyloxycarbonyl (Boc), then compound A-VIa is treated with HCl to give compound A-2HC1, which is converted to compound A-HCl. In some embodiments, the AHCl compound or the A-2HC1 compound is treated with an appropriate base, such as sodium hydroxide, sodium carbonate, sodium bicarbonate, or the like, to provide Compound A (free base form).

In some embodiments, the crystalline form of Compound A, Compound A-HCl, or Compound A-2HC1 is isolated directly from the reaction mixture. In some embodiments, the crystalline form of Compound A, Compound A-HCl, or Compound A-2HC1 was formed by recrystallization from a suitable solvent. Suitable solvents include, without limitation, water, methanol, ethanol, ethyl acetate, isopropyl acetate, pentane, hexanes, heptane, or combinations thereof.

- 14 042196

Synthesis of 6-bromo-3-chloroquinolin-4-ol (A-II).

A-1 A-II

In some embodiments, Compound A-II is prepared by chlorination of Compound A-I. In some embodiments, chlorination is performed by treating compound A-I with a chlorinating agent such as N-chlorosuccinimide (NCS), trichloroisocyanuric acid (TCA), sulfuryl chloride, chlorine, sodium hypochlorite, calcium hypochlorite, hypochlorous acid, or 2,3,4,5,6, 6-hexachloro-2,4-cyclohexadien-1-one or the like. In some embodiments, chlorination was performed by treating Compound A-I with N-chlorosuccinimide (NCS).

In some embodiments, the chlorination is carried out in a suitable solvent such as acetic acid, water, ethanol, methanol, toluene, dichloromethane, tetrahydrofuran, dioxane, N,N-dimethylformamide, or the like.

In some embodiments, the chlorination is carried out in acetic acid. In some embodiments, 1 volume of acetic acid refers to an amount that is the same volume as the reactants in the reaction, i. A-I and/or NCS. In some embodiments, the reaction is carried out in about 5, 7.5, 10, 12.5, 15, or 20 volumes of acetic acid. In some embodiments, the reaction is carried out in about 10 volumes of acetic acid. In some embodiments, the reaction is carried out in about 12.5 volumes of acetic acid. In some embodiments, the reaction is carried out in about 20 volumes of acetic acid.

In some embodiments, the chlorination was carried out at an elevated temperature. In some embodiments, the reaction temperature is from about 40°C to about 150°C. In some embodiments, the reaction temperature was from about 40°C to about 120°C. In some embodiments, the reaction temperature was from about 40°C to about 100°C. In some embodiments, the reaction temperature was from about 40°C to about 80°C. In some embodiments, the reaction temperature is from about 40°C to about 60°C. In some embodiments, the reaction temperature was from about 45°C to about 55°C. In some embodiments, the reaction temperature was about 40°C, 45°C, 50°C, 55°C, or 60°C. In some embodiments, the reaction temperature was about 50°C.

Synthesis of 4,6-dibromo-3-chloroquinoline (A-III).

A-ll A-H1a

In some embodiments, the hydroxyl group of compound A-II is converted to a halogen atom (X is Br or Cl).

In some embodiments, the halogenation is a bromination. In some embodiments, the halogenation is chlorination.

In some embodiments, Compound A-II has been brominated with phosphorus tribromide (PBr ₃ ), phosphorus oxybromide (POBr ₃ ), hydrobromic acid, bromine, dibromotriphenylphosphorane, or the like. In some embodiment, Compound A-II was brominated with PBr ₃ . In some embodiments, if Compound A-II is brominated, the product is Compound A-III.

In some embodiments, Compound A-II is chlorinated with POCl ₃ , thionyl chloride, or the like. In some embodiments, A-II was chlorinated with POCl ₃ .

In some embodiments, the halogenation reaction is carried out in a suitable solvent. In some embodiments, a suitable solvent is acetonitrile, water, ethanol, isopropanol, dichloromethane, toluene, N,N-dimethylformamide, acetic acid, acetone, or the like. In some embodiments, the halogenation solvent is DMF. In some embodiments, the reaction temperature ranged from 0°C to 30°C. In some embodiments, the reaction temperature ranged from 0° C. to room temperature. In some embodiments, the reaction is started at 0°C and warmed to room temperature.

- 15 042196

Synthesis of N-protected 1-(6-bromo-3-chloroquinolin-4-yl)piperidine (A-IVa).

In some embodiments, Compound A-Iva was prepared from Compound A-III. In some embodiments, Compound A-IVa is prepared from Compound A-III by reacting Compound A-III with N-protected 4-aminopiperidine in the presence of a base in a suitable solvent.

In some embodiments, the amino protecting group is a carbamate (such as 9-fluorenylmethyl carbamate (Fmoc), t-butyl carbamate (Boc), benzyl carbamate (Cbz), or the like), an amide (such as acetyl, trifluoroacetyl, or the like) , phthalimide, benzyl, trityl, benzylideneamine, tosyl, or the like. In some embodiments, the amino protecting group is a carbamate. In some embodiments, the amino protecting group is Boc. In some embodiments, if the protecting group is Boc, the product is Compound A-IV.

In some embodiments, the base is a non-nucleophilic base. In some embodiments, the base is an amine base. In some embodiments, the base is an amine base such as triethylamine (TEA), diisopropylethylamine (DIPEA), 1,8-diazabicycloundec-7-ene (DBU), 1,2,2,6,6-pentamethylpiperidine, tributylamine, or t .P. In some embodiments, the base is DIPEA.

In some embodiments, the base is an inorganic base. In some embodiments, the base is a carbonate (MCO3) or bicarbonate (MHCO3) base, where M is sodium, potassium, or cesium. In some embodiments, the base is an inorganic base such as sodium bicarbonate, Na ₂ CO ₃ , K ₂ CO ₃ , Cs ₂ CO ₃ , or the like.

In some embodiments, a suitable reaction solvent is N,N-dimethylformamide, N,N-dimethylacetamide, dimethylsulfoxide, dichloromethane, chloroform, dioxane, tetrahydrofuran, toluene, acetonitrile, ethanol, isopropanol, or the like. In some embodiments, the reaction solvent is dimethyl sulfoxide.

In some embodiments, the reaction temperature is from about 0°C to about 250°C, from about 50°C to about 200°C, or from about 100°C to about 180°C. In some embodiments, the reaction temperature is from about 120°C to about 150°C. In some embodiments, the reaction temperature is about 100, 105, 11, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, or 180°C.

Alternative synthesis of N-protected 1-(6-bromo-3-chloroquinolin-4-yl)piperidine (compound A-IVa).

In some embodiments, Compound A-Iva was prepared from Compound 3. In some embodiments, Compound A-IVa was prepared from Compound 3 by reacting Compound 3 with N-protected 4-aminopiperidine in the presence of a base in a suitable solvent to give Compound 4a followed by chlorination compounds 4a. In some embodiments, if the protecting group is Boc, compound 4a is compound 4.

In some embodiments, the amino protecting group is a carb

- 16 042196 mate (such as 9-fluorenylmethyl carbamate (Fmoc), tert-butyl carbamate (Boc), benzyl carbamate (Cbz) or the like), amide (such as acetyl, trifluoroacetyl or the like), phthalimide, benzyl, trityl, benzylideneamine, tosyl or the like. In some embodiments, the amino protecting group is a carbamate. In some embodiments, the amino protecting group is Boc. In some embodiments, if the protecting group is Boc, the product is compound A-IV.

In some embodiments, the base is a non-nucleophilic base. In some embodiments, the base is an amine base. In some embodiments, the base is an amine base such as triethylamine (TEA), diisopropylethylamine (DIPEA), 1,8-diazabicycloundec-7-ene (DBU), 1,2,2,6,6-pentamethylpiperidine, tributylamine, or t .P. In some embodiments, the base is DIPEA. In some embodiments, the base is an inorganic base. In some embodiments, the base is a carbonate (MCO ₃ ) or bicarbonate (MHCO ₃ ) base, where M is sodium, potassium, or cesium. In some embodiments, the base is an inorganic base such as sodium bicarbonate, Na ₂ CO ₃ , K ₂ CO ₃ , Cs ₂ CO ₃ , or the like. In some embodiments, a suitable solvent for the reaction is N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, dichloromethane, chloroform, dioxane, tetrahydrofuran, toluene, acetonitrile, ethanol, isopropanol, or the like. In some embodiments, the solvent for the reaction is N,N-dimethylformamide. In some embodiments, the reaction temperature is from about 0°C to about 250°C, from about 50°C to about 200°C, or from about 100°C to about 180°C. In some embodiments, the reaction temperature is from about 120°C to about 150°C. In some embodiments, the reaction temperature is about 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, or 180°C.

In some embodiments, Compound A-Iva was prepared by chlorination of Compound 4a. In some embodiments, chlorination was performed by treating compound 4a with a chlorinating agent such as N-chlorosuccinimide (NCS), trichloroisocyanuric acid (TCA), sulfuryl chloride, chlorine, sodium hypochlorite, calcium hypochlorite, hypochlorous acid, or 2,3,4,5,6, 6-hexachloro-2,4-cyclohexadien-1-one or the like. In some embodiments, chlorination was performed by treating compound 4a with N-chlorosuccinimide (NCS). In some embodiments, the chlorination is carried out in a suitable solvent such as acetic acid, water, ethanol, methanol, toluene, dichloromethane, tetrahydrofuran, dioxane, N,N-dimethylformamide, or the like. In some embodiments, the chlorination is carried out in toluene. In some embodiments, the chlorination was carried out at an elevated temperature. In some embodiments, the reaction temperature is from about 40°C to about 150°C. In some embodiments, the reaction temperature is from about 40°C to about 120°C. In some embodiments, the reaction temperature is from about 40°C to about 100°C. In some embodiments, the reaction temperature is from about 50°C to about 80°C. In some embodiments, the reaction temperature was about 70°C.

Synthesis of (3-cyano-2-hydroxyphenyl)boronic acid (compound 1).

CN _ CN

2) Borylation

3) Withdrawal

_2a protecting groups

Also disclosed in the present invention is a two-step process for synthesizing compound 1 wherein B is a boronic acid or a boronic acid ester, the method comprising the steps of (1) protecting the hydroxyl group of compound 2 with an appropriate protecting group (PG) to form compound 2a ; (2) reacting compound 2a with a borylating agent under suitable reaction conditions; and (3) removing the protecting group (PG') to give Compound 1.

In some embodiments, a suitable protecting group (PG') is methoxymethyl (MOM), ethoxyethyl (EE), methoxypropyl (MOP), benzyloxymethyl (BOM), 2methoxyethoxymethyl (MEM), benzyl (Bn), p-methoxybenzyl (PMB), 2-naphthylmethyl (Nap), methyl (Me), allyl, tetrahydropyranyl (THP), acetyl (Ac), benzoyl (Bz), 2,2,2-trichloroethylcarbonyl (Troc), [2(trimethylsilyl)ethoxy]methyl, trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS), tert-butyldimethylsilyl (TBDMS), or tert-butyldiphenylsilyl (TBDPS).

In some embodiments, a suitable protecting group (PG') is methoxymethyl, 2-methoxyethoxymethyl, benzyl, p-methoxybenzyl, methyl, allyl, tetrahydropyr

- 17 042196 nyl, [2-(trimethylsilyl)ethoxy]methyl, trimethylsilyl, triethylsilyl, triisopropylsilyl, t-butyldimethylsilyl or t-butyldiphenylsilyl. In some embodiments, a suitable protecting group (PG') is methoxymethyl or 2-methoxyethoxymethyl. In some embodiments, a suitable protecting group (PG) is methoxymethyl. In some embodiments, a suitable protecting group (PG) is acetyl.

In some embodiments, deprotection in step (3) was performed with different reagents depending on what the protecting group was. In some embodiments, deprotection in step (3) was performed by treatment with hydrochloric acid, hydrobromic acid, acetic acid, trifluoroacetic acid, tosylic acid, ZnBr ₂ , hydrogen over Pd/C, 2,3-dichloro-5,6-dicyano- 1,4-benzoquinone (DDQ), boron tribromide, boron trichloride, trimethylsilyl iodide, Pd(PPh ₃ ) ₄ , tetra-n-butylammonium fluoride (TBAF) or HF-pyridine. In some embodiments, deprotection in step (3) was performed by treatment with hydrochloric acid, hydrobromic acid, acetic acid, trifluoroacetic acid, or tosylic acid. In some embodiments, the deprotection of step (3) was performed by treatment with hydrochloric acid.

In some embodiments, the boryling agent is pinacolborane, catecholborane, bis(neopentylglycolate)diborone, bis(pinacolato)diborone, bis(hexylene glycolate)diborone, bis(catecholate)diborone, tetrahydroxydiborone, trimethoxyboron, triisopropoxyboron, 4,4,5,5- tetramethyl 1,3,2-dioxaborolane, 4,6,6-trimethyl-1,3,2-dioxaborinane, diisopropylaminoborane, bis(neopentylglycolate)diborone, bis(catecholato)diborone or bis(pinacolato)diborone. In some embodiments, suitable reaction conditions (2) include the use of halometal exchange reagents. In some embodiments, suitable reaction conditions (2) include the use of halogen-metal exchange reagents selected from Grignard reagents and alkyllithium reagents. In some embodiments, suitable reaction conditions (2) include the use of isopropyl magnesium chloride in tetrahydrofuran. In some embodiments, the borylating agent is triisopropoxyboron and suitable reaction conditions (2) include the use of isopropyl magnesium chloride in tetrahydrofuran. In some embodiments, suitable reaction conditions (2) include the use of transition metal mediated reaction conditions. In some embodiments, suitable reaction conditions (2) include the use of palladium metal mediated reaction conditions.

In some embodiments, B is boronic acid or an ester of boronic acid. In some embodiments, B is

In some embodiments, B is boronic acid. According to , ⁱⁿ H but in some embodiments, B is ·

In some embodiments, B is a boronic acid ester. In some embodiments, B is

In some embodiments, the method further includes an additional step (4) wherein the boronic acid or boronic acid ester is converted to

trifluoroborate. In such embodiments, B is

In some embodiments, such a transformation is achieved by treating the boron-containing compound with KHF2.

Synthesis of tert-butyl ( 1 - (6- (3 - cyano-2-hydroxyphenyl) -3 - (3,5-difluorophenyl) quinolin-4-yl) piperidin4-yl) carbamate (compound A-VI).

- 18 042196

In some embodiments, compound A-VIa was synthesized from compound AIVa. In some embodiments, Compound A-VIa is prepared by conducting two consecutive Suzuki reactions of Compound A-IVa. In some embodiments, compound A-IVa is reacted in the first Suzuki reaction with compound 1, a suitable palladium catalyst, a suitable ligand, and a suitable base in a suitable solvent to form compound A-Va. In some embodiments, compound A-Va is reacted in a second Suzuki reaction with 3,5-difluorophenylboronic acid, a suitable palladium catalyst, a suitable ligand, and a suitable base in a suitable solvent to give compound A-VIa. In some embodiments, Compound A-Va is isolated prior to the second Suzuki reaction. In some embodiments, compound A-Va was not isolated prior to the second Suzuki reaction. If PG is Boc, then an A-Va compound is an A-V compound.

In some embodiments, suitable bases in Suzuki reactions include amine bases and inorganic bases. Suitable amine bases for Suzuki reactions include, but are not limited to, triethylamine, diisopropylethylamine, 1,2,2,6,6pentamethylpiperidine, tributylamine, 1,8-diazabicycloundec-7-ene (DBU), or the like. Suitable inorganic bases for Suzuki reactions include, but are not limited to, NaOAc, KOAc, Ba(OH) ₂ , Li ₂ CO ₃ , Na ₂ CO ₃ , K ₂ CO ₃ , Cs ₂ CO ₃ , Na ₃ PO ₄ , K ₃ PO ₄ or the like. In some embodiments, the base in each Suzuki reaction is the same. In some embodiments, the base in each Suzuki reaction is not the same. In some embodiments, about 1, 2, 3, 4, 5, or 6 equivalents of base are used in the Suzuki reaction(s). In some embodiments, if compound A-Va was not isolated prior to the second Suzuki reaction, no additional base was added to the reaction.

In some embodiments, the amount of palladium for each Suzuki reaction is less than about 0.05 equiv. In some embodiments, the amount of palladium for each Suzuki reaction is from about 0.05 equiv. up to about 0.2 equiv. In some embodiments, the amount of palladium for each Suzuki reaction is about 0.05, 0.1, 0.15, or 0.2 equiv. In some embodiments, the amount of palladium for each Suzuki reaction is the same. In some embodiments, the amount of palladium for each Suzuki reaction is not the same.

In some embodiments, a suitable ligand is used for the palladium catalyst. In some embodiments, the ligand is a phosphine ligand. In some embodiments, the ligand is an aliphatic phosphine ligand such as trimethylphosphine, tricyclohexylphosphine, tri-t-butylphosphine, or the like. In some embodiments, the ligand is an aromatic phosphine such as XPhos, SPhos, JohnPhos, Amphos, triphenylphosphine, methyldiphenylphosphine, or the like. In some embodiments, the ligand is a phosphite ligand such as trimethyl phosphite, triphenyl phosphite, or the like. In some embodiments, the ligand is a bisphosphine ligand such as diphenylphosphinomethane (dppm), diphenylphosphinoethane (dppe), 1,1'bis(diphenylphosphino)ferrocene (dppf), or the like. In some embodiments, the ligand for each Suzuki reaction is the same. In some embodiments, is

- 19 042196 the gand for each Suzuki reaction is not the same.

In some embodiments, the palladium source in one or both of the Suzuki reactions is a Pd(O) source such as Pd ₂ (dba) ₃ , Pd(PPh ₃ ) ₄ , or the like. In some embodiments, the palladium source in one or both of the Suzuki reactions is a Pd(II) source such as PdCl ₂ , Pd(OAc) 2 , PdCl ₂ (PPh ₃ ) ₂ , PdCl 2 (dppf)-DCM, PdCl 2 (Amphos) or etc. In some embodiments, the source of palladium for each Suzuki reaction is the same. In some embodiments, the source of palladium for each Suzuki reaction is different.

In some embodiments, the solvent system used in one or both of the Suzuki reactions is a single solvent. In some embodiments, the solvent system used in one or both of the Suzuki reactions is a mixture of co-solvents. In some embodiments, the solvent system used in one or both of the Suzuki reactions is toluene, DMF, acetonitrile, EtOH, IPA, THF, dioxane, water, or mixtures thereof. In some embodiments, the solvent system used in each Suzuki reaction is the same. In some embodiments, the solvent system used in each Suzuki reaction is not the same.

In some embodiments, the temperature used in one or both of the Suzuki reactions is from about 50°C to 150°C, preferably from about 60°C to 120°C. In some embodiments, the temperature used in one or both of the Suzuki reactions is from about 80°C to 85°C. In some embodiments, the temperature used in one or both of the Suzuki reactions is 90°C to 100°C. In some embodiments, each Suzuki reaction was run at the same temperature. In some embodiments, each Suzuki reaction is run at a different temperature.

In some embodiments, Compound A-Va is isolated between reactions. In some embodiments, Compound A-Va was not isolated between reactions. In some embodiments, each Suzuki reaction was performed in the same reaction vessel. In some embodiments, each Suzuki reaction was performed in the same reaction vessel without isolating compound A-Va. In some embodiments, each Suzuki reaction was carried out in the same solvent. In some embodiments, each Suzuki reaction was run with the same base.

In one aspect, two Suzuki reactions were carried out sequentially in the same reaction vessel without isolating intermediate A-Va and without removing solvent, wherein after the first reaction was completed, the reaction vessel was allowed to cool before adding the second boronic acid, and then the reaction vessel was heated for the second Suzuki reaction. In some embodiments, the second palladium source and/or ligand was added with the second boronic acid. In some embodiments, no additional source of palladium and/or ligand was added with the second boronic acid. In some embodiments, an additional base was added to the reaction with the second boronic acid. In some embodiments, no additional base was added to the reaction with the second boronic acid.

In some embodiments, the progress of one or both of the Suzuki reactions was monitored by HPLC or by TLC.

In some embodiments, the solvent system used is 10:1 dioxane:water. In some embodiments, the base used is K ₂ CO ₃ and 4 equiv. grounds. In some embodiments, the first palladium source is PdCl ₂ (dppf)-CH ₂ Cl ₂ . In some embodiments, the second palladium source is PdCl2(Amphos). In some embodiments, the first Suzuki reaction takes place at about 80-85°C. In some embodiments, the second Suzuki reaction takes place at about 90-100°C.

In some embodiments, Compound A-VIa contains a detectable amount of residual palladium as an impurity. If PG is Boc, then the A-VIa compound is the A-VI compound.

In some embodiments, the isolated compound A-VI product contains a detectable amount of unreacted compound A-V. In some embodiments, a sample of Compound A-VI contains a detectable amount of an impurity selected from

- 20 042196

In some embodiments, Compound A-VI is purified by recrystallization.

- 21 042196

In some embodiments, Compound A-VI is heated in a suitable solvent or mixture of solvents for an appropriate period of time. In some embodiments, the purity of Compound A-VI was improved in this manner (see Table 1 below). In some embodiments, such a recrystallization/suspension method removes or reduces the amount of residual palladium in Compound A-VI samples.

Table 1

HPLC solids purity Crystallization (RC) / suspension Purity (%) Unreacted compound A-V (%) Other impurities (%) RC from MTBE 96.22 1.05 2.72 RC from RA 97.10 0.68 2.22 RC from EA 95.47 2.18 2.35 RC from Meon 92.96 1.60 1.86 Precipitation from DCM/pet. ether 96.28 1.09 1.86 Suspension in ACN 95.61 1.32 2.51 RC from THF/water 93.72 2.27 3.40 Precipitation from DMF/water 94.35 2.17 2.85 K. t. suspension in toluene 96.36 0.41 3.24 Hot suspension in toluene 97.57 0.16 2.28 HPLC solids purity Crystallization (RC) / suspension Purity (%) Unreacted compound A-V (%) Other impurities (%) From DCM/MTBE 96.78 0.80 2.42 Hot slurry from IP Ac 98.13 0.00 1.87 RC from THF/ Petr. ether 98.24 0.16 1.59 Hot suspension in Meon 96.30 0.35 3.35

^a The purity of starting compound A-VI for all RC/slurry studies was 96.18%

Due to the fact that a transition metal catalyst was used in the synthesis methods described above, purification steps were performed to reduce the amount of palladium in the product. Purification steps to reduce the amount of palladium in the product were carried out so that the active pharmaceutical ingredients matched the technical specifications of palladium. (Guideline on the Specification Limits for Residues of Metal Catalysts European Medicines Agency Pre-authorisation Evaluation of Medicines for Human Use, London, January 2007, Doc. Ref. CPMP/SWP/QWP/4446/00 corr.). In some embodiments, purification steps to reduce the amount of palladium in the product include, but are not limited to, treatment with solid trimercaptotriazine (TMT), polystyrene-bonded TMT, mercaptoporous polystyrene-bonded TMT, polystyrene-bonded ethylenediamine, activated carbon, glass bead sponges, Smopex™ , silicon binding scavengers, thiol-derivatized silica gel, N-acetylcysteine, h-Bu _3P , crystallization, extraction, 1-cysteine, n-Bu ₃ P/lactic acid (Garrett et al., Adv. Synth. Catal. 2004, 346 , 889-900). In some embodiments, activated carbon includes, without limitation, DARCO® KB-G, DARCO® KB-WJ. In one aspect, silicon-binding scavengers include, but are not limited to

means silica gel.

In some embodiments, the purification steps to reduce the amount of palladium include the use of activated carbon, derivatized silica gel (eg, thiolderivated silica gel), or combinations thereof.

In some embodiments, Compound A-VI is further treated with a metal scavenger to remove residual palladium. In some embodiments, the metal scavenger comprises SiO ₂ , carbon, L-cysteine aqueous solution, Silicycle metal scavenger,

Si-thiol, SiliaBond DMT or SiliaBond cysteine. In some embodiments, the absorbent loading (w/w) is 1:3, 1:2, or 1:1. In some embodiments, the metal scavenger is Si-thiol.

In some embodiments, the crude Compound A-VI, as isolated from the reaction, is treated with a metal scavenger. In some embodiments, the recrystallized Compound A-VI is treated with a metal scavenger. In some of these embodiments, palladium levels have been reduced sufficiently to be undetectable.

In some embodiments, the presence of residual heavy metal impurities (eg, palladium) was determined using methods known in the art. In some embodiments, the presence of residual heavy metal impurities (eg, palladium) was determined using inductively coupled plasma mass spectrometry (ICPMS). In some embodiments, the presence of residual heavy metal impurities (e.g., palladium) was determined using the techniques described in U.S. Pharmacopeia General Chapter <231> Heavy Metals.

Synthesis of 3-(4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl)-2-hydroxybeshonitrile, di-HCl salt (compound A-2HC1).

Connection A-2HCI

In some embodiments, compound A-VI is treated with hydrochloric acid in a suitable solvent to give compound A-2HCl. In some embodiments, a suitable solvent is isopropyl alcohol (IPA), methyl tert-butyl ether (MTBE), toluene, ethyl acetate, isopropyl acetate, water, or mixtures thereof. In some embodiments, a suitable solvent is isopropyl alcohol, ethyl acetate, or isopropyl acetate. In some embodiments, a suitable solvent is IPA.

Preparation of compound A-HCl from compound A-2HCl.

A-2HCI connection A-HCI connection

In some embodiments, compound A-2HCl is dissolved in a suitable solvent and treated with aqueous ammonia to give compound A-HCl. In some embodiments, a suitable solvent is IPA, water, methyl acetate, or combinations thereof. In some embodiments, the solvent is water. In some embodiments, the solvent is IPA:water in a ratio of 9:1, 8:2, 7:3, 6:4, or 5:5. In some embodiments, the solvent is methyl acetate:water in a ratio of 9:1, 8:2, 7:3, 6:4, or 5:5. In some embodiments, additional sodium bicarbonate is added to the reaction mixture. In some embodiments, the solvent volume is 5, 10, 15, 20, or greater than 20 volumes.

In some embodiments, the aqueous ammonia source is saturated ammonium chloride (28-30%). In other embodiments, the aqueous ammonia source is 25%. In some embodiments, about 1 equivalent of ammonia was added. In some embodiments, less than 1 equivalent of ammonia is added, such as 0.8 equivalents. In some embodiments, more than one equivalent of ammonia is added, such as 1.25 equivalents. In some embodiments, the amount of ammonia added was determined by monitoring the pH of the solution. In some embodiments, the pH is adjusted to about 4-6 by adding ammonia, bicarbonate, or hydroxide.

- 23 042196

In some embodiments, the pH is adjusted by adding ammonia. In some embodiments, the pH is adjusted to about 4.5-4.7 by adding ammonia.

In some embodiments, the reaction mixture is heated to about 30, 35, 40, 45, 50, 55, or 60°C. In some embodiments, the reaction mixture is heated to about 45°C. In some embodiments, the reaction mixture was heated prior to the addition of ammonia.

In some embodiments, water is added to the reaction mixture while heating. In some embodiments, water is added to the reaction mixture at the end of the reaction. In some embodiments, water is added over about 2 hours to the reaction mixture.

In some embodiments, the slurry is filtered to isolate the AHCl compound or solvate thereof.

In some embodiments, ammonia is not necessary to convert compound A-2HC1 to compound A-HCl. In some embodiments, the A2HCl compound is stirred in about 10, about 20, about 30, about 40, or about 50 volumes of water. In some embodiments, Compound A-2HCl is stirred in about 20 to about 50 volumes of water. In some embodiments, sodium bicarbonate is added to the aqueous mixture to adjust the pH. In some embodiments, the A-HCl compounds are isolated from solids filtration from this aqueous mixture.

Preparation of compound A from compound A-2HCl.

Compound A-2HCI Compound A

In some embodiments, the free base of Compound A is obtained by treating Compound A-2HCl with a suitable base in a suitable solvent. In some embodiments, a suitable base is sodium hydroxide, sodium bicarbonate, or the like. In some embodiments, a suitable solvent is water. In some embodiments, the solid is filtered from the mixture to isolate the free base of Compound A.

Obtaining compound A-HCl from compound A.

Compound A Compound A-HCI

In some embodiments, compound A-HCl is formed by treating the free base with about 1 equivalent of HCl in a suitable solvent. In some embodiments, a suitable solvent is methanol, ethanol, isopropyl alcohol, acetone, methyl acetate, ethyl acetate, isopropyl acetate, tetrahydrofuran, tetrahydropyran, water, acetone, or combinations thereof. In some embodiments, the HCl source is HCl in IPA, HCl in toluene, HCl in MTBE, or HCl in water. In some embodiments, the AHCl compound is isolated by solids filtration from the reaction mixture.

Additional connections

According to another aspect described in the present invention, there is provided a compound of formula (I), or a pharmaceutically acceptable salt, a pharmaceutically acceptable solvate, a diastereomeric mixture, or an enantiomer thereof:

- 24 042196

Formula (I) where

R ^a is F, Cl or -CH3;

R ^b is hydrogen, F, Cl, -CH3, -CN, -OH or -OCH3;

R ^B is unsubstituted or substituted phenyl or unsubstituted or substituted pyridinyl, where if R ^B is substituted then R ^B is substituted by R ^c and R ^d ;

R ^c is hydrogen, F, Cl, Br, -CH3, -CN, -OH, -OCH3, -C(=NOCH3)H or -C(=NOH)H;

R ^d is -OH or -NH2; and each R ^f is hydrogen or F.

In some embodiments, a compound of formula (I) is provided that has the structure of formula (Ia), or a pharmaceutically acceptable salt, solvate, diastereomeric mixture, single enantiomers, or prodrug thereof:

Formula (1a)

According to some variants of implementation of R ^B is described in table. 2.

In some embodiments, the compounds described herein have the following structure, or a pharmaceutically acceptable salt, solvate, diastereomeric mixture, or individual enantiomers thereof:

Where

R ^a , R ^b and R ^B are described in table. 2.

In some embodiments, R ^a , R ^b , and R ^B are described in Table 1. 2.

Any combination of the groups described above for various variables is contemplated herein. Throughout the description, the groups and their substituents are selected by those skilled in the art to ensure stable moieties and compounds.

Given as an example of the compounds of formula (I) include the compounds described in table. 2.

- 25 042196

table 2

conn. no. ^Rf R ^{B Ra Rb} 1-1 Η he 1 z ^z k/ ^0H QT F F 1-2 Η ξ /Υ^ ^νη 2 ί τ Ή''/I F -SNz 1-3 Η ξ /L,nh ₂ Γ τ Cl F 1-4 Η 1 7 J 4^μ ^νη 2 F -SNz 1-5 Η G ( 7 1 4^/ ^ΝΗ 2 Cl F 1-6 Η OH 1 _Ά^/ΝΗ ₂ ί Υ F F 1-7 Η 1 ( τ 1 4^ ^ΝΗ 2 F H 1-8 Η CH3 ^L^NH ₂ f Υ F H

- 26 042196

Connections in the table. 2 named as

1-1: 2-{4-[(4a8,8a8)-octahydro-1H-pyrido[3,4-b][1,4]oxazin-6-yl]-3-(3,5difluorophenyl)quinoline-6 -yl } -6-[(1 E)-(hydroxyimino)methyl]phenol;

1-2: 2-{4-[(4a8,8a8)-octahydro-1H-pyrido[3,4-b][1,4]oxazin-6-yl]-3-(3-fluoro-5methylphenyl)quinoline -6-yl}-4-[( 1 E)-(methoxyimino)methyl]pyridine-3-amine;

1-3: 2-{4-[(4a8,8a8)-octahydro-1H-pyrido[3,4-b][1,4]oxazin-6-yl]-3-(3-chloro-5fluorophenyl)quinoline -6-yl}-4-[( 1 E)-(methoxyimino)methyl]pyridine-3-amine;

1-4: 2-{4-[(4a8,8a8)-octahydro-1H-pyrido[3,4-b][1,4]oxazin-6-yl]-3-(3-fluoro-5methylphenyl)quinoline -6-yl }-3-amino pyride in-4-carbonitryl;

1-5: 2-{4-[(4a8,8a8)-octahydro-1H-pyrido[3,4-b][1,4]oxazin-6-yl]-3-(3-chloro-5fluorophenyl)quinoline -6-yl}-3-aminopyridine-4-carbonitryl;

1-6: 2-{4-[(4a8,8a8)-octahydro-1H-pyrido[3,4-b][1,4]oxazin-6-yl]-3-(3,5d ifluorophenyl)quinolin- 6-yl}-4-[(1 E)-(hydroxyimino)methyl]pyridine-3-amine;

1-7: 2-{4-[(4a8,8a8)-octahydro-1H-pyrido[3,4-b][1,4]oxazin-6-yl]-3-(3fluorophenyl)quinolin-6-yl }-3-aminopyridine-4-carbonitrile and

1-8: 2-{4-[(4a8,8a8)-octahydro-1H-pyrido[3,4-b][1,4]oxazin-6-yl]-3-(3fluorophenyl)quinolin-6-yl }-4-methylpyride in-3-amine.

In some embodiments, the present invention provides a pharmaceutically acceptable salt of a compound as described in Table 1. 2.

Also provided in the present invention is compound 1-9: 3-[4-(4-aminopiperidin-1-yl)-5fluoro-3-(3-fluoro-5-methylphenyl)quinolin-6-yl]-2-hydroxybenzonitrile or its pharmaceutically acceptable salt.

Also provided in the present invention is compound 1-10: 3-[4-(4-aminopiperidin-1-yl)-7fluoro-3-(3-fluoro-5-methylphenyl)quinolin-6-yl]-2-hydroxybenzonitrile or its pharmaceutically acceptable salt.

In one aspect, the compounds described herein are in the form of pharmaceutically acceptable salts. In addition, active metabolites of such compounds that exhibit the same type of activity are included within the scope of this disclosure. In addition, the compounds described in the present invention may exist in unsolvated as well as solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like. Solvated forms of the compounds provided in the present invention are also considered as disclosed in the present invention.

In some embodiments, the compounds described herein are prepared as described in Scheme B.

- 27 042196

Scheme B

a) cyclic amine, ΕΐβΝ; b) R ^b B(OH)2, Pd; c) R ^A B(OH)2, Pd; d) deprotection; f) bis(pinacolato)diborone, Pd, X-Phos, KOAc; f) R ^B X, Pd

In some embodiments, Compound I is converted to Compound II by nucleophilic substitution with an appropriate chlorine cyclic amine in the presence of Et ₃ N or DIEA. Selective Suzuki-Miyaura reactions with R ^B B(OH)2 or its boronic acid ester gave compound IV, which can also be prepared in a two-step sequence by formation of a boronic acid ester (compound III) followed by selective Suzuki-Miyaura reactions with R ^B X (X=Cl, Br or I). Suzuki-Miyaura reactions with R ^A (OH) ₂ or its boronic acid ester followed by deprotection of all protecting groups using appropriate deprotection methods, e.g. acidic, resulted in compound V.

In some embodiments, the compounds obtained by the above methods were obtained as racemic or diastereomeric mixtures. In some embodiments, racemic mixtures of compounds have been resolved to obtain optically pure (or optically enriched) isomers using conventional chiral separation techniques such as chiral HPLC, chiral supercritical fluid chromatography (SFC), pseudo-moving bed chromatography (SMB), and the like. .

In some embodiments, diastereomeric mixtures of compounds have been resolved to obtain optically pure (or optically enriched) isomers using crystallization techniques or conventional non-chiral chromatographic techniques such as silica gel chromatography or chiral chromatographic techniques such as chiral HPLC, chiral supercritical liquid chromatography (SFC), pseudo-moving bed chromatography (SMB), and the like.

In some embodiments, the compounds described herein were synthesized as described in the examples.

As used herein, pharmaceutically acceptable refers to a substance, such as a carrier or diluent, that does not interfere with the biological activity or properties of the compound and is relatively non-toxic, i. the substance was administered to the subject without causing undesirable biological effects or without interacting in an inappropriate manner with any of the components of the composition in which it is contained.

The term pharmaceutically acceptable salt refers to the form of the therapeutically effective agent that consists of the cationic form of the therapeutically active agent in combination with a suitable anion, or in alternative embodiments, the anionic form of the therapeutically active agent in combination with a suitable cation. Handbook of Pharmaceutical Salts: Properties, Selection and Use. International Union of Pure and Applied Chemistry, Wiley-VCH 2002. S.M. Berge, L.D. Bighley, D.C. Monkhouse, J. Pharm. sci. 1977, 66, 1-19. P. H. Stahl and C G. Wermuth, editors, Handbook of Pharmaceutical Salts: Properties, Selection and Use, Weinheim/Zurich: Wiley-VCH/VHCA, 2002. Pharmaceutical salts are typically more soluble and more rapidly soluble in the stomach and gastric juice than non-ionic species, and thus usable as solid dosage forms. Moreover, since their solubility is often a function of pH, selective dissolution in one or another part of the gastrointestinal tract is possible and this ability can be controlled as

- 28 042196 one aspect of the modes of prolonged and sustained release. Also, since the salt-forming molecule can be in equilibrium with the neutral form, passage through biological membranes can be adjusted.

In some embodiments, pharmaceutically acceptable salts are prepared by reacting a compound of the present invention with an acid. In some embodiments, a compound (ie, the free base form) disclosed herein is basic and reacts with an organic acid or inorganic acid. Inorganic acids include, without limitation, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, and metaphosphoric acid. Organic acids include, without limitation, 1-hydroxy-2-naphthoic acid; 2,2 dichloroacetic acid; 2-hydroxyethanesulfonic acid; 2-oxoglutaric acid; 4acetamidobenzoic acid; 4-aminosalicylic acid; acetic acid; adipic acid; ascorbic acid (L); aspartic acid (L); benzenesulfonic acid; benzoic acid; camphoric acid (+); camphor-10-sulfonic acid (+); capric acid (decanoic acid); caproic acid (hexanoic acid); caprylic acid (octanoic acid); carbonic acid; cinnamic acid; citric acid; cyclamic acid; dodecylsulfuric acid; ethane 1,2-disulfonic acid; ethanesulfonic acid; formic acid; fumaric acid; galactic acid; gentisic acid; glucoheptonic acid (D); gluconic acid (D); glucuronic acid (D); glutamic acid; glutaric acid; glycerophosphoric acid; glycolic acid; hippuric acid; isobutyric acid; lactic acid (DL); lactobionic acid; lauric acid; maleic acid; malic acid (-L); malonic acid; mandelic acid (DL); methanesulfonic acid; naphthalene-1,5-disulfonic acid; naphthalene-2-sulfonic acid; nicotinic acid; butyric acid; oxalic acid; palmitic acid; pamoic acid; phosphoric acid; propionic acid; pyroglutamic acid (-L); salicylic acid; sebacic acid; stearic acid; succinic acid; sulfuric acid; tartaric acid (+L); thiocyanic acid; toluenesulfonic acid (para) and undecenoic acid.

In some embodiments, a compound disclosed herein is obtained as the chloride salt, sulfate salt, bromide salt, mesylate salt, maleate salt, citrate salt, or phosphate salt.

In some embodiments, pharmaceutically acceptable salts are prepared by reacting a compound disclosed in the present invention with a base. In some embodiments, a compound disclosed in the present invention is acidic and reacts with a base. In such cases, the acidic proton of the compound disclosed in the present invention is replaced by a metal ion, such as a lithium, sodium, potassium, magnesium, calcium or aluminum ion. In some cases, the compounds described in the present invention interact with an organic base, such as, without limitation, ethanolamine, diethanolamine, triethanolamine, tromethamine, meglumine, N-methylglucamine, dicyclohexylamine, tris(hydroxymethyl)methylamine. In other instances, the compounds described herein form salts with amino acids such as, but not limited to, arginine, lysine, and the like. Suitable inorganic bases used to form salts with compounds that include an acidic proton include, but are not limited to, aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, sodium hydroxide, lithium hydroxide, and the like. In some embodiments, the compounds of the present invention are provided as the sodium salt, calcium salt, potassium salt, magnesium salt, meglumine salt, N-methylglucamine salt, or ammonium salt.

It should be understood that reference to a pharmaceutically acceptable salt includes solvent addition products. In some embodiments, the solvates contain either stoichiometric or non-stoichiometric amounts of solvent and are formed during the crystallization process with pharmaceutically acceptable solvents such as water, ethanol, and the like. Hydrates are formed if the solvent is water, or alcoholates are formed if the solvent is alcohol. Solvates of the compounds described in the present invention for the sake of convenience were obtained or formed during the methods described in the present invention. In addition, the compounds of the present invention optionally exist in unsolvated as well as solvated forms.

Therapeutic agents that have been administered to mammals, such as humans, should be prepared following regulatory guidelines. These government-regulated guidelines are called the Goods Manufacturing Practices (GMP). GMP guidelines set out acceptable levels of contamination of active therapeutic agents, such as, for example, the amount of residual solvent in the final product. Preferred solvents are those that are suitable for use in GMP facilities and comply with safety issues. Solvent categories are defined in, for example, International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH), Impuri- 29 042196 ties: Guidelines for Residual Solvents, Q3C(R3), (November 2005).

Solvents are categorized into three classes.

Class 1 solvents are toxic and should be avoided. Class 2 solvents are solvents that are limited in use during the manufacture of a therapeutic agent. Class 3 solvents are solvents with low toxic potential and low risk to human health. Data for class 3 solvents means they are less toxic in acute or short term studies and negative in genotoxicity studies.

Class 1 solvents to avoid include: benzene; carbon tetrachloride; 1,2-dichloroethane; 1,1-dichloroethene and 1,1,1-trichloroethane.

Examples of class 2 solvents are: acetonitrile, chlorobenzene, chloroform, cyclohexane, 1,2-dichloroethene, dichloromethane, 1,2-dimethoxyethane, N,N-dimethylacetamide, N,Ndimethylformamide, 1,4-dioxane, 2-ethoxyethanol, ethylene glycol , formamide, hexane, methanol, 2methoxyethanol, methylbutylketone, methylcyclohexane, N-methylpyrrolidine, nitromethane, pyridine, sulfolane, tetralin, toluene, 1,1,2-trichloroethene, and xylene.

Class 3 solvents that have low toxicity include: acetic acid, acetone, anisole, 1-butanol, 2-butanol, butyl acetate, tert-butyl methyl ether (MTBE), cumene, dimethyl sulfoxide, ethanol, ethyl acetate, ethyl ether, ethyl formate , formic acid, heptane, isobutyl acetate, isopropyl acetate, methyl acetate, 3-methyl-1-butanol, methyl ethyl ketone, methyl isobutyl ketone, 2-methyl-1-propanol, pentane, 1-pentanol, 1-propanol, 2-propanol, propyl acetate, and tetrahydrofuran.

Residual solvents in active pharmaceutical ingredients (APIs) come from the manufacture of APIs. In some cases, solvents are not completely removed by practical manufacturing techniques. Appropriate choice of solvent for API synthesis can enhance yield or determine characteristics such as crystal form, purity, and solubility. Thus, the solvent is a critical parameter in the synthesis process.

In some embodiments, compositions containing Compound A, or a pharmaceutically acceptable salt thereof, include an organic solvent(s). In some embodiments, compositions containing Compound A, or a pharmaceutically acceptable salt thereof, include a residual amount of organic solvent(s). In some embodiments, compositions containing Compound A, or a pharmaceutically acceptable salt thereof, include a residual Class 3 solvent. In some embodiments, the Class 3 solvent is selected from the group consisting of acetic acid, acetone, anisole, 1-butanol, 2-butanol, butyl acetate, t-butyl methyl ether, cumene, dimethyl sulfoxide, ethanol, ethyl acetate, ethyl ether, ethyl formate, formic acid , heptane, isobutyl acetate, isopropyl acetate, methyl acetate, 3-methyl-1-butanol, methyl ethyl ketone, methyl isobutyl ketone, 2-methyl-1-propanol, pentane, 1-pentanol, 1-propanol, 2-propanol, propyl acetate, and tetrahydrofuran. In some embodiments, the Class 3 solvent is selected from ethyl acetate, isopropyl acetate, t-butyl methyl ether, heptane, isopropanol, and ethanol.

In some embodiments, compositions containing Compound A, or a pharmaceutically acceptable salt thereof, include a detectable amount of an organic solvent. In some embodiments, the pharmaceutically acceptable salt of Compound A is the HCl salt (ie Compound A-HCl). In some embodiments, the organic solvent is a class 3 solvent.

In other embodiments, compositions are provided comprising Compound A, or a pharmaceutically acceptable salt thereof, wherein the composition contains a detectable amount of solvent that is less than about 1%, wherein the solvent is selected from acetone, 1,2dimethoxyethane, acetonitrile, ethyl acetate, tetrahydrofuran, methanol, ethanol, heptane and 2 propanol. In a further embodiment, compositions are provided comprising Compound A, or a pharmaceutically acceptable salt thereof, wherein the composition contains a detectable amount of solvent that is less than about 5000 ppm. In another further embodiment, compositions are provided comprising Compound A, wherein the detectable amount of solvent is less than about 5000 ppm, less than about 4000 ppm, less than about 3000 ppm, less than about 2000 ppm, less than about 1000 ppm, less than about 500 ppm or less than about 100 ppm.

The methods and compositions described in the present invention include the use of N-oxides (if necessary) or pharmaceutically acceptable salts of compounds having the structure disclosed in the present invention, as well as active metabolites of such compounds having one type of activity.

In some embodiments, sites on organic radicals (eg, alkyl groups, aromatic rings) of the compounds disclosed in the present invention are susceptible to various metabolic reactions. Inclusion of appropriate substituents on organic radicals will shorten, minimize or eliminate such a metabolic pathway.

- 30 042196

In particular embodiments, an appropriate substituent for reducing or desensitizing an aromatic ring to metabolic reactions is, by way of example only, a halogen, deuterium, alkyl group, haloalkyl group, or deuteroalkyl group.

In another embodiment, the compounds described herein are labeled isotopically (eg, with a radioisotope) or by other means, including, without limitation, the use of chromophores or fluorescent moieties, bioluminescent labels, or chemiluminescent labels.

The compounds described herein include isotopically labeled compounds which are identical to the compounds set forth in the various formulas and structures provided herein, but with one or more atoms replaced by an atom with an atomic mass or mass number different from the atomic mass or mass number commonly found in nature. Examples of isotopes that can be included in the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, sulfur, fluorine, chlorine, iodine, phosphorus, such as, for example, ² H, ³ H, ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁸ O, ¹⁷ O, ³⁵ S, ¹⁸ F, ³⁶ Cl, ¹²³ I, ¹²⁴ I, ¹²⁵ I, ¹³¹ I, ³² P, and ³³ P. In one aspect, the isotopically labeled compounds described herein, for example , compounds in which radioactive isotopes such as ³ H and ¹⁴ C are incorporated are useful in drug and/or substrate tissue distribution assays. In one aspect, substitution with isotopes such as deuterium provides certain therapeutic benefits resulting from greater resistance to metabolic inactivation, such as, for example, increased in vivo half-life or altered metabolic pathways to reduce unwanted metabolites or reduced dosage.

In some embodiments, one or more hydrogen atoms on Compound A are replaced by deuterium. In some embodiments, deuterium substitution allows certain therapeutic benefits resulting from greater metabolic stability, such as, for example, increased in vivo half-life or reduced dosage requirement.

In one aspect, a compound of the following structure is described:

where each R is independently selected from hydrogen or deuterium, or a pharmaceutically acceptable salt thereof.

In some embodiments, the pharmaceutically acceptable salt of the compound is the HCl salt. In some embodiments, the pharmaceutically acceptable salt of the compound is the DCl salt.

In some embodiments, the compounds disclosed herein have one or more stereocenters, and each stereocenter exists independently in any of the R or S configurations. For example, in some embodiments, a compound disclosed herein exists in the R configuration if one stereocenter is present. In other embodiments, a compound disclosed herein exists in an S configuration if one stereocenter is present. In some embodiments, a compound disclosed herein exists in an RR configuration if two stereocenters are present. In some embodiments, a compound disclosed herein exists in an RS configuration if two stereocenters are present. In some embodiments, a compound disclosed herein exists in an SS configuration if two stereocenters are present. In some embodiments, a compound disclosed herein exists in an SR configuration if two stereocenters are present.

The compounds provided herein include all diastereomeric, individual enantiomeric, atropisomeric and epimeric forms, as well as their respective mixtures. The compounds and methods provided herein include all cis, trans, syn, anti, opposite (E), and one side (Z) isomers, as well as their respective mixtures.

Individual stereoisomers were obtained, if necessary, by methods such as stereoselective synthesis and/or separation of stereoisomers using chiral chromatographic columns, or separation of diastereomers using either non-chiral or chiral chromatographic columns, or crystallization and recrystallization in an appropriate solvent or in a mixture of solvents. custodians. In certain embodiments, the compounds disclosed herein are prepared as their individual stereoisomers by reacting a racemic mixture of the compound with an optically active resolving agent to form a diastereomeric compound/salt pair, separating the diastereomers and recovering the optically pure individual enantiomers. In some embodiments, cleavage of the individual enantiomers of the compounds disclosed herein is carried out using covalent diastereomeric derivatives of the compounds described herein. In another embodiment, the diastereomers of the compounds disclosed herein are separated by separation/cleavage methods based on differences in solubility. In other embodiments, separation of stereoisomers of the compounds disclosed in the present invention was carried out by chromatography or by formation of diastereomeric salts and separation by recrystallization or chromatography, or any combination thereof. Jean Jacques, Andre Collet, Samuel H. Wilen, Enantiomers, Racemates and Resolutions, John Wiley And Sons, Inc., 1981. In some embodiments, stereoisomers are prepared by stereoselective synthesis.

Separation of individual enantiomers from a racemic mixture is possible using chiral supercritical liquid chromatography (SFC) or chiral high performance liquid chromatography (HPLC). In some embodiments, the enantiomers described herein are separated from each other using chiral SFC or chiral HPLC. In some embodiments, compounds disclosed herein that include one or more chiral centers (e.g., compounds disclosed herein that include a trans-octahydro-III-pyrido[3,4-b]morpholine- 6yl), separated into individual enantiomers using chiral SFC or chiral HPLC. A wide range of conditions and suitable columns are available.

Daicel's polysaccharide chiral stationary phases (CSPs) are present among the columns used for chiral SFC separations. According to some Daicel embodiments, CHIRALPAK and CHIRALCEL HPLC analytical fixed and coated columns can be used for SFC analysis.

In some embodiments, SFC column suitability screening was performed on four main stationary phases (CHIRALPAK IA, IB, IC and ID) and four main coated columns (CHIRALPAK AD and AS and CHIRALCEL OD and OJ) with varying concentrations of organic modifier. A range of column phases are available, including but not limited to OD and OJ, OX and OZ chlorinated phases, and a range of additional CHIRALCEL cellulose based phases including OA, OB, OC, OF, OG and OK.

Non-limiting examples of chiral selectors for use in separating enantiomers include tris(3,5-dimethylphenylcarbamate) amylose, tris(3,5dimethylphenylcarbamate) cellulose, tris(3,5-dichlorophenylcarbamate) cellulose, tris(3-chlorophenylcarbamate) amylose , amylose tris(3,5-dichlorophenylcarbamate), amylose tris(3-chloro,4-methylphenylcarbamate), amylose tris((S)-alpha-methylbenzylcarbamate), amylose tris(5-chloro-2-methylphenylcarbamate), cellulose tris( 4methylbenzoate), tris(4-chloro-3-methylphenylcarbamate) cellulose, and tris(3-chloro-4methylphenylcarbamate) cellulose.

Non-limiting examples of chiral columns contemplated for use in separating enantiomers include CHIRALPAK IA SFC, CHIRALPAK AD-H SFC, CHIRALPAK IB SFC, CHIRALCEL OD-H SFC, CHIRALPAK IC SFC, CHIRALPAK ID SFC, CHIRALPAK IE SFC, CHIRALPAK IF SFC , CHIRALPAK AZ-H SFC, CHIRALPAK AS-H SFC, CHIRALPAK AY-H SFC, CHIRALCEL OJ-H SFC, CHIRALCEL OX-H SFC, and CHIRALCEL OZ-H SFC.

In accordance with some embodiments, the similarity and position of the substituents in the compounds described herein help to minimize undesirable activity. For example, in accordance with some embodiments, the unwanted activity includes unwanted hERG inhibition. In some embodiments, the presence of a hydroxyl group and an adjacent cyano group on the aromatic ring significantly attenuates undesired inhibition of hERG compared to the absence of both groups, the presence of a hydroxyl group without an adjacent cyano group, or the presence of a cyano group without an adjacent hydroxyl group. For example, in some embodiments, a significant reduction in unwanted hERG inhibition is observed when R ^B is substituted or unsubstituted 2-hydroxy-3-cyanophenyl.

According to subsidiary or additional embodiments, the compounds described herein are metabolized upon administration to the body in need of production of a metabolite, which is then used to obtain the desired effect, including the desired therapeutic effect.

The term metabolite of a compound disclosed herein means a derivative of the compound that is formed during the metabolism of the compound. The term active metabolite refers to the biologically active derivative of a compound that is formed during metabolism

- 32 042196 connections. As used herein, the term metabolizable refers to the sum of processes (including, but not limited to, hydrolysis reactions and reactions catalyzed by enzymes) in which a particular substance is altered by the body. Thus, enzymes can provide certain structural changes to the compound. For example, cytochrome P450 catalyses a range of oxidative and reductive reactions, while uridine diphosphate glucuronyltransferases catalyze the transfer of an activated glucuronic acid molecule to aromatic alcohols, aliphatic alcohols, carboxylic acids, amines, and free sulfhydryl groups. Metabolites of the compounds disclosed herein are optionally identified either by administering the compounds to the recipient and analyzing tissue samples from the recipient, or by incubating the compounds with liver cells in vitro and analyzing the resulting compounds.

Unless otherwise noted, the following terms used in the present invention are characterized by the definitions below. The use of the term including, as well as other forms such as include, includes, and included, is not limited. The section headings used herein are for organizational purposes only and are not intended to limit the subject matter discussed.

The term halo or alternatively halogen or halide means fluorine, chlorine, bromine or iodine. In some embodiments, the halogen is fluorine, chlorine, or bromine.

The term bond or simple bond refers to a chemical bond between two atoms or two fragments if the atoms connected by the bond are considered to be part of a larger substructure. In one aspect, if the group described herein is a bond, there is no control group, thus allowing bond formation between the remaining established groups.

The term fragment refers to a particular segment or functional group of a molecule. Chemical moieties are often shown as chemical entities embedded in or attached to a molecule.

Used in the present invention, the term acceptable in relation to the composition, composition or ingredient means not having a permanent adverse effect on the general health of the subject who was treated.

As used herein, the term modulate means interacting with a target either directly or indirectly to change the activity of the target, including, by way of example only, increasing the activity of the target, inhibiting the activity of the target, limiting the activity of the target, or prolonging the activity of the target.

As used herein, the term modulator refers to a molecule that interacts with a target, either directly or indirectly. Interactions include, without limitation, agonist, partial agonist, inverse agonist, antagonist, destructor, or combinations thereof. In some embodiments, the modulator is an agonist.

As used herein, the terms administer, administer process, administer, and the like. refer to methods that can be used to effect delivery of compounds or compositions to a desired site of biological action. Such methods include, without limitation, oral routes, intraduodenal routes, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intravascular, or infusion), topical, and rectal administration. Those skilled in the art are aware of administration techniques that can be used with the compounds and methods described herein. In some embodiments, the compounds and compositions described herein are administered orally.

As used herein, the terms co-administration or the like encompass the administration of selected therapeutic agents to a single patient and include treatment regimens in which the agents are administered by the same or different route of administration, or at the same or different times.

As used herein, the terms effective amount or therapeutically effective amount refer to a sufficient amount of the agent or compound to be administered that will alleviate to some extent one or more symptoms of the disease or condition being treated. The result includes a reduction and/or amelioration of the signs, symptoms, or causes of a disease, or some other desired change in the biological system. For example, an effective amount for therapeutic applications is the amount of a composition containing a compound disclosed herein required to provide a clinically significant reduction in the symptoms of a disease. The appropriate effective amount in any particular case is optionally determined using techniques such as a dose escalation study.

As used herein, the terms enhance or amplify means to increase or prolong either the efficacy or the duration of the desired effect. Thus, in relation to the enhancing effect of therapeutic agents, the term enhancement refers to the ability

- 33 042196 increase or prolong either the effectiveness or the duration of the effect of other therapeutic agents in the system. As used herein, the term enhancing effective amount refers to an amount suitable to enhance the effect of another therapeutic agent in the desired system.

As used herein, the term pharmaceutical combination means a product that is the result of mixing or combining more than one active ingredient and includes both fixed and non-fixed combinations of active ingredients. The term fixed combination means that the active ingredients, for example, both the compound disclosed in the present invention, or a pharmaceutically acceptable salt thereof, and the co-agent, are administered to the patient simultaneously in the form of a single substance or dosage. The term non-fixed combination means that the active ingredients, for example, both a compound disclosed in the present invention or a pharmaceutically acceptable salt thereof, and a joint agent, are administered to the patient in the form of separate substances either simultaneously, in parallel, or sequentially without any specific time intervals, at This introduction provides an effective content of the two compounds in the patient's body. The latter also applies to cocktail therapy, for example, the administration of three or more active ingredients.

The terms product and kit are used interchangeably.

The term subject or patient includes mammals.

Examples of mammals include, without limitation, any member of the class

Mammals: humans, non-human primates such as chimpanzees and other narrow-nosed monkeys, as well as other types of monkeys; farm animals such as cattle, horses, sheep, goats, pigs; pets such as rabbits, dogs and cats; laboratory animals, including rodents such as rats, mice, guinea pigs, and the like. In one aspect, the mammal is a human.

As used herein, the terms treat, process of treatment, or treatment include reducing, alleviating, or alleviating at least one symptom of a disease or condition, preventing additional symptoms, suppressing a disease or condition, such as delaying the development of a disease or condition, ameliorating a disease or condition, providing regression of the disease or condition, amelioration of the condition caused by the disease or condition, or suspension of the symptoms of the disease or condition, either prophylactically and/or therapeutically.

Pharmaceutical compositions

In some embodiments, the compounds described herein are formulated into pharmaceutical compositions. Pharmaceutical compositions are conventionally formulated using one or more pharmaceutically acceptable inactive ingredients which facilitate the processing of the active compounds into pharmaceutically useful formulations. The appropriate composition depends on the chosen route of administration. Brief descriptions of the pharmaceutical compositions described herein are, for example, Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, H.A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, N.Y., 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins 1999), incorporated herein by reference for such disclosure.

In some embodiments, the compounds described herein are administered either alone or in combination with pharmaceutically acceptable carriers, adjuvants, or diluents in a pharmaceutical composition. Administration of the compounds and compositions described herein may be by any method that delivers the compounds to the site of action. These methods include, without limitation, delivery by enteral routes (including oral, gastric or duodenal feeding tube, rectal suppository and rectal enema), parenteral routes (by injection or infusion, including intra-arterial, intracardiac, intradermal, intraduodenal, intramedullary, intramuscular, intraosseous, intraperitoneal, intrathecal, intravascular, intravenous, intravitreal, epidural and subcutaneous), inhalation, transdermal, transmucosal, sublingual, buccal and local (including cutaneous, skin, enema, eye drops, ear drops, intranasal, vaginal) administration, although the most appropriate route may depend, for example, on the condition and impairment of the recipient. By way of example only, the compounds described herein may be administered locally to the site in need of treatment, such as by local infusion during surgery, topical application, such as creams or ointments, injection, catheter, or implant. The introduction can also be carried out by direct injection at the site of the diseased tissue or organ.

In some embodiments, pharmaceutical compositions suitable for oral administration are presented as discrete units such as capsules, sachets, or tablets, each containing a predetermined amount of the active ingredient, as

- 34 042196 powder or granules, in the form of a solution or suspension in an aqueous medium or non-aqueous medium, or in the form of an oil-in-water liquid emulsion or a water-in-oil liquid emulsion. In some embodiments, the active ingredient is presented as a bolus, electuary, or paste.

Pharmaceutical compositions that can be used orally include tablets, hard capsules made from gelatin, and soft, stoppered capsules made from gelatin and a plasticizer such as glycerol or sorbitol. Tablets can be obtained by compression or molding, optionally with one or more auxiliary ingredients. Compressed tablets can be prepared by compressing in a suitable machine the active ingredient in free flowing form such as powder or granules, optionally mixed with binders, inert diluents or lubricants, surfactants or dispersants. Molded tablets can be prepared by shaping in a suitable machine a mixture of a powdered compound moistened with an inert liquid diluent. In some embodiments, tablets are coated or scored and formulated to provide slow or controlled release of the active ingredient from them. All formulations for oral administration should be in dosages suitable for such administration. Hard capsules may contain the active ingredients in admixture with an excipient such as lactose, binders such as starches and/or lubricants such as talc or magnesium stearate, and optionally stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids such as fixed oils, liquid paraffin or liquid polyethylene glycols. In some embodiments, stabilizers are added. Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol and/or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures. Dyes or pigments may be added to tablets or dragee coatings to identify or characterize different dose combinations of the active compound.

In some embodiments, the pharmaceutical compositions are formulated for parenteral administration by injection, eg, bolus injection or continuous infusion. Compositions for injection may be presented in unit dosage form, for example, in ampoules or in multi-dose containers, with an added preservative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles and may contain adjuvants such as suspending, stabilizing and/or dispersing agents. The compositions may be presented in single-dose or multi-dose containers, such as sealed ampoules and vials, and may be stored in powder form or in a freeze-dried (lyophilized) state requiring only the addition of a sterile liquid carrier, such as saline or sterile pyrogen-free water immediately before use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets of the type previously described.

Pharmaceutical compositions for parenteral administration include aqueous and non-aqueous (oily) sterile injectable solutions of the active compounds, which may contain antioxidants, buffers, bacteriostatics, and dissolved components that render the formulation isotonic with the blood of the intended recipient; as well as aqueous and non-aqueous sterile suspensions, which may include suspending agents and thickeners. Suitable lipophilic solvents or carrier vehicles include fatty oils such as sesame oil or synthetic fatty acid esters such as ethyl oleate or triglycerides or liposomes. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol or dextran. Optionally, the suspension may also contain suitable stabilizers or agents which increase the solubility of the compounds to ensure that highly concentrated solutions are obtained.

Pharmaceutical compositions may also be formulated as a depot formulation. Such long acting formulations may be administered by implantation (eg subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, compounds can be formulated with suitable polymeric or hydrophobic materials (eg, as an emulsion in a suitable oil), or ion exchange resins, or as sparingly soluble derivatives, eg, as a sparingly soluble salt.

For buccal or sublingual administration, the compositions may take the form of tablets, lozenges, lozenges, or gels formulated by conventional means. Such compositions may contain the active ingredient in a flavor base such as sucrose and gum arabic or gum tragacanth.

Pharmaceutical compositions can be administered topically, ie by non-systemic administration. This includes applying a compound of the present invention topically to the epidermis or buccal cavity and instilling such compound into the ear, eye and nose such that the compound does not enter the bloodstream to a significant extent. In contrast, systemic administration refers to oral, intravenous, intraperitoneal, and intramuscular administration.

Pharmaceutical compositions suitable for topical administration include liquid or semi-liquid preparations suitable for penetrating the skin to the site of inflammation, such as gels, liniments, lotions, creams, ointments or patches and drops suitable for administration to the eye, ear or nose. . For topical administration, the active ingredient may comprise from 0.001% to 10% w/w, for example 1 to 2% by weight of the formulation.

Pharmaceutical compositions for administration by inhalation are conveniently delivered by insufflator, pressurized aerosol cans, or other suitable aerosol spray delivery means. The pressurized cylinders may contain a suitable propellant such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or other suitable gas. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to deliver a metered amount. Alternatively, for administration by inhalation or insufflation, the pharmaceutical preparations may take the form of a dry powder composition, for example a powder mixture of a compound and a suitable powder base such as lactose or starch. The powder composition may be presented in unit dosage form, for example, in capsules, cartridges, gelatin or blister packs, from which the powder may be administered using an inhaler or insufflator.

It should be appreciated that, in addition to the ingredients specifically mentioned above, the compounds and compositions described herein may include other agents conventional in the art relevant to the type of formulation in question, for example, agents suitable for oral administration, may include flavoring agents.

Dosing routes and treatment regimens

In one embodiment, the compounds disclosed herein, or a pharmaceutically acceptable salt thereof, are used in the preparation of medicaments for the treatment of diseases or conditions in a mammal for which modulation of somatostatin activity would be effective. Methods for treating any of the diseases or conditions described herein in a mammal in need of such treatment include administering to said mammal pharmaceutical compositions that include at least one compound disclosed herein, or a pharmaceutically acceptable salt thereof, an active metabolite, prodrug or pharmaceutically acceptable solvate, in therapeutically effective amounts.

In certain embodiments, compositions containing the compound(s) described herein are administered for prophylactic and/or therapeutic treatments. In some therapeutic applications, the compositions are administered to a patient already suffering from a disease or condition in an amount sufficient to cure or at least partially relieve at least one of the symptoms of the disease or condition. Amounts effective for such use will depend upon the severity and course of the disease or condition, prior therapy, the health of the patient, weight and response to drugs, and the judgment of the attending physician.

Therapeutically effective amounts are optionally determined by methods including, but not limited to, dose escalation and/or dose variation clinical trials.

In prophylactic applications, compositions containing the compounds described herein are administered to a patient susceptible to or otherwise at risk for a particular disease, disorder or condition. Such an amount is defined as a prophylactically effective amount or dose. In this application, the exact amounts also depend on the patient's health, weight, and the like. When used in patients, effective amounts for such use will depend upon the severity and course of the disease or condition, prior therapy, the health of the patient, weight and response to drugs, and the judgment of the attending physician. In one aspect, prophylactic treatments comprise administering to a mammal that has previously experienced at least one symptom of the disease being treated and is currently in remission, a pharmaceutical composition comprising a compound disclosed in the present invention, or a pharmaceutically acceptable salt thereof, to prevent relapse. symptoms of a disease or condition.

In certain embodiments, if the patient's condition does not improve, then at the physician's discretion, the administration of the compounds is chronic, that is, for an extended period of time, including throughout the life of the patient, to alleviate or otherwise control or limit the symptoms of the disease or condition of the patient. .

After the patient's condition improves, if necessary, a maintenance dose is administered. Then, in specific embodiments, the dosage or frequency of administration, or both, is reduced as a function of symptoms to a level at which improvement in the disease, disorder, or condition is maintained. In certain embodiments, however, intermittent treatment is required on a long-term basis for any recurrence of symptoms.

The amount of a given agent that corresponds to such an amount varies depending on

- 36 042196 on factors such as the specific compound, the condition of the disease and its severity, the characteristics (eg, weight, sex) of the subject or recipient in need of treatment, but is nevertheless determined according to the specific circumstances associated with this case, in including, for example, the specific agent to be administered, the route of administration, the condition to be treated, and the subject or recipient to be treated.

However, in general, doses used to treat an adult typically range from 0.01 mg to 2000 mg per day. In one embodiment, the desired dose is conventionally presented as a single dose or divided doses administered simultaneously or at appropriate intervals, eg two, three, four or more sub-doses per day.

In one embodiment, suitable daily dosages for a compound disclosed herein, or a pharmaceutically acceptable salt thereof, are from about 0.01 to about 50 mg/kg body weight. In some embodiments, daily dosages or amounts of the active ingredient in dosage form are below or above the ranges set forth herein, depending on a number of variables in an individual treatment regimen. In various embodiments, daily and unit dosages will vary depending on a number of variables, including, without limitation, the activity of the compound used, the disease or condition being treated, the route of administration, the requirements of the individual subject, the severity of the disease or condition being treated, and the judgment of the practitioner.

The toxicity and therapeutic efficacy of such therapeutic regimens is determined by standard pharmaceutical procedures in cell cultures or experimental animals, including, without limitation, determination of LD ₅₀ and ED ₅₀ . The ratio of dose to toxic and therapeutic effects is the therapeutic index and is expressed as the ratio of LD ₅₀ to ED ₅₀ . In certain embodiments, data obtained from cell culture assays and animal studies is used in formulating a range of therapeutically effective daily dosages and/or a therapeutically effective number of unit dosages for use in mammals, including humans. In some embodiments, the daily dosage of the compounds described herein is in a range of circulating concentrations that include the ED ₅₀ with minimal toxicity. In certain embodiments, the daily dosage range and/or the number of unit dosages varies within this range depending on the dosage form used and the route of administration used.

According to any of the above aspects, further embodiments are provided wherein an effective amount of a compound disclosed in the present invention, or a pharmaceutically acceptable salt thereof:

(a) administered to the mammal systemically; and/or (b) administered orally to the mammal; and/or (c) administered intravenously to a mammal; and/or (d) administered to the mammal by injection; and/or (e) administered topically to the mammal; and/or (f) administered to the mammal non-systemically or locally.

According to any of the above aspects, further embodiments are provided comprising single administrations of an effective amount of the compound, including further embodiments wherein (i) the compound is administered once a day; or (ii) the compound is administered to the mammal several times over the course of one day.

According to any of the above aspects, further embodiments are provided comprising multiple administrations of an effective amount of a compound, including further embodiments wherein (i) the compound is administered continuously or intermittently as a single dose;

(ii) time intervals between multiple injections are 6 hours;

(ii i) the compound is administered to the mammal every 8 hours;

(iv) the compound is administered to the mammal every 12 hours;

(v) the compound is administered to the mammal every 24 hours.

In additional or alternative embodiments, the method comprises a drug holiday in which the administration of the compound is temporarily suspended or the dose of the compound to be administered is temporarily reduced; at the end of drug holidays, dosing of the compound is resumed. According to one embodiment, the length of the drug holiday varies from 2 days to 1 year.

Complex treatment

In some instances, it may be appropriate to administer at least one compound disclosed herein, or a pharmaceutically acceptable salt thereof, in combination with one or more other therapeutic agents.

In one embodiment, the therapeutic efficacy of one of the compounds

- 37 042196 described herein, is increased by the introduction of an excipient (ie, the excipient itself has minimal therapeutic benefit, but in combination with another therapeutic agent, the overall therapeutic benefit for the patient is enhanced). Or, in some embodiments, the benefit experienced by the patient is enhanced by administering one of the compounds described herein with another agent (which also includes a therapeutic regimen) that also has a therapeutic benefit.

In one particular embodiment, a compound disclosed herein, or a pharmaceutically acceptable salt thereof, is co-administered with a second therapeutic agent, wherein the compound disclosed herein, or a pharmaceutically acceptable salt thereof, and the second therapeutic agent modulate various aspects of the disease, disorder, or condition being treated, thereby providing a greater overall benefit than administering either therapeutic agent alone.

In any case, with respect to the disease, disorder, or condition being treated, the overall benefit experienced by the patient is simply the sum of the two therapeutic agents, or the patient experiences a synergistic effect.

For the combination therapeutics described herein, the dosages of the co-administered compounds will vary depending on the type of drug being co-administered, the particular drug being used, the disease or condition being treated, and the like. In additional embodiments, when administered in conjunction with one or more other therapeutic agents, a compound provided herein is administered either simultaneously with one or more other therapeutic agents or sequentially.

In the case of combination therapeutics, multiple therapeutics (one of which is one of the compounds described herein) are administered in any order or even simultaneously. If the administration is simultaneous, then several therapeutic agents, by way of example only, are provided in a single unitary form or in several forms (for example, as a single pill or as two separate pills).

Compounds disclosed in the present invention, or a pharmaceutically acceptable salt thereof, as well as combination therapeutics, are administered before, during, or after the onset of a disease or condition, and the timing of administration of the composition containing the compound varies. Thus, in one embodiment, the compounds described herein are used prophylactically and administered chronically to subjects prone to developing conditions or diseases to prevent the onset of the disease or condition. In another embodiment, the compounds and compositions are administered to the subject during or as soon as possible after the onset of symptoms. In specific embodiments, a compound described herein is administered as soon as possible after the onset of a disease or condition, identified or suspected, and for the length of time necessary to treat the disease. In some embodiments, the length of time required for treatment varies and the duration of treatment is set to meet the specific needs of each subject.

Examples

Abbreviations:

BINAP: (2,2'-bis(diphenylphosphino)-1,1'-binaphthyl);

DCE: 1,2-dichloroethane;

DCM: dichloromethane;

D.I.: deionized; DIEA or DIPEA: diisopropylethylamine;

EtOAc: ethyl acetate;

EtOH: ethanol;

Equiv.: equivalents, typically molar equivalents;

HATU: 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate;

IPA: isopropyl alcohol;

IPAc: isopropyl acetate;

MeOAc: methyl acetate;

NBS: N-bromosuccinimide;

NCS: N-chlorosuccinimide;

OPA: phosphoric acid;

PTS: para-toluenesulfonic acid;

Pd(Amphos)Cl ₂ : bis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II);

Pd ₂ dba ₃ : tris(dibenzylideneacetone)dipalladium (0);

Pd(dppf)Cl ₂ : [1.1'-bis(diphenylphosphino)ferrocene]dichloropalladium (II);

(pinB) ₂ : bis(pinacolato)diborone;

k.t. or RT: room temperature;

- 38 042196

Rt: retention time;

SFC: supercritical liquid chromatography;

SST: somatostatin;

SSTR: somatostatin receptor;

TEA: trimethylamine;

TFA: trifluoroacetic acid;

THF: tetrahydrofuran;

Volume: volume, typically used for the reaction, the volume or ratio of solvents;

Xphos: 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl;

clock: clock;

h or hour: hour.

The following examples are presented for illustrative purposes only and do not limit the scope of the claims presented herein.

Example 1 Synthesis of 3-bromo-2-(methoxymethoxy)benzonitrile (compound 2b).

CN ^he MOMCI, DIPEA, DCM

25°С, 4 h Br

CN

OMOM

Br

2b

Diisopropylethylamine (114 ml, 1.3 equiv.) was slowly added to a solution of nitrile compound 2 (100 g, 1 equiv.) in CH ₂ Cl ₂ (1 L, 10 vol.) at 0°C and stirred for 30 min. Chloromethyl methyl ether (MOMCl) (46 ml, 1.2 equiv.) was then slowly added while maintaining the internal temperature at 0 to 5°C. The reaction mixture was then allowed to warm to rt. and stirred for 4 hours until completion of the reaction was observed by TLC. The reaction mixture was cooled to 0° C. and quenched with DI water (300 ml, 3 vol) and the layers were separated. The aqueous layer was extracted with CH ₂ Cl ₂ (300 ml, 3 vol.) and the combined organic layer was washed with water and brine, concentrated on a rotary vacuum evaporator to obtain 115 g of the crude product as a brown oil. The crude product was purified on a SiO ₂ plug and eluted with 10% ethyl acetate and petroleum ether (20 vol.). Only one fraction was collected, evaporated under vacuum and dried under high vacuum to give 86 g (70%) of compound 2b as a colorless oil, the purity of which was 99.96% (HPLC-AUC).

Example 2 Synthesis of (3-cyano-2-hydroxyphenyl)boronic acid (compound 1a).

OMOM

i) 'PrMgCI, THF _he -5 to 0°C, 10 min (H0) ₂ V CN ) ('PrO) ₃ V 0°C to rt, 16 h iii) 3 n HCI, from 0 °C to k.t., 4 h

1a

A solution of 'PrMgCl in THF (2M in THF, 340 ml, 2.2 equiv.) was added slowly to a solution of compound 2b (75 g, 1 equiv.) in THF (1.12 L, 15 vol.) while maintaining the internal temperature at - 5 to 5°C and stirred for 10 min. Triisopropyl borate (180 ml, 2.5 equiv.) was then added while maintaining the internal temperature at -5 to 3°C. The reaction mixture was then allowed to warm to room temperature and stirred for 18 hours until completion of the reaction was observed by TLC. The reaction mixture was then cooled to -10°C and extinguished by the slow addition of 3 n HCl (620 ml, 6 equiv.) at -10°C. The mixture was stirred for 3 h at rt. and was extracted with ethyl acetate (525 ml, 5 vol.) and the aqueous layer was extracted with ethyl acetate (225 ml, 3 vol.). The combined organic layer was washed successively with DI water (3x3 vol.), brine (525 ml, 3 vol.) and concentrated in vacuo to give the crude material as a sticky solid. The crude material was stirred in petroleum ether (525 ml, 5 vol) for 30 min and the resulting solids were filtered, washed with petroleum ether (150 ml, 2 vol) and dried under vacuum to give 35 g (70%) of the boronic acid compound 1a as an off-white solid which was 97.91% pure (HPLC-AUC).

- 39 042196

Example 3 Synthesis of 2-bromo-6-cyanophenylacetate (compound 2c).

3-Bromo-2-fluoro-benzonitrile (25 g, 1.0 equiv.), potassium acetate (5 equiv.) were mixed in DMSO (7 vol.) and heated to 90-95°C for 48 hours. IPC observed 0.38% 3-bromo-2-fluorobenzonitrile and 96.5% compound 2. The reaction mixture was cooled to 25-30°C and quenched with purified water (25 vol. water). The pH was then adjusted to 3-4 using 6N HCl. The resulting reaction mixture was diluted with MTBE (10 vol.). The organic layer was separated and the aqueous layer was extracted with 10 vol. MTBE. The combined organic layers were washed with water (10 vol x 3) and concentrated to vol. 2 was passed through DCM (3 vol.) and concentrated again to 2 vol. before diluting with DCM (8 vol.) to obtain a crude solution of compound 2. This solution was used in the following method without further purification.

The crude solution of compound 2 was mixed with acetic anhydride (1.3 equiv.), DMAP (0.1 equiv.) at 25-30°C for 2 h with stirring. By IPC, 0.8% of compound 2 and 94.8% of compound 2c were observed. The reaction mass was diluted with purified water (10 vol.), stirred for 30 min. The organic layer was separated. The aqueous layer was extracted with 2 vol. DCM. The combined organic layer was washed with water (8 vol.x2). Charcoal (10%) was added to the organic layer and stirred for 1 hour before being filtered through a Celite layer. The filtrate was then concentrated to vol. 2, passed through 3 about. and 2 about. n-heptane subsequently before cooling to rt. and stirred for 1 h at 5-10°C. The product was isolated by filtration as a solid (25.5 g, 85% yield over two steps, HPLC 96.8% purity).

Example 4 Alternative synthesis of 2-hydroxy-3-cyanopotassium phenyl trifluoroborate (compound 1c).

Compound 2c (20 g, 1.0 equiv.) was mixed with KOAc (3.0 equiv.), bis(pinacolato)diborone (1.2 equiv.) in 2-methyl-THP. The mixture was stirred and degassed by bubbling N ₂ before adding Pd(dppf)Cl ₂ -DCM (0.025 equiv.). The resulting mixture was heated to 80-85° C. for 16 hours. 0.3% starting material and 92% compound 1b were observed by IPC. The reaction mixture was cooled to 2530°C and filtered through a pad of celite. The Celite layer was washed with MTBE (5 vol.). The combined filtrate was concentrated to 2 vol. and passed through MTBE to 2 vol. The resulting solution was diluted with MTBE (10 vol.) and stirred for 1 h at ambient temperature. The slurry was again filtered through a pad of celite and the pad of celite was washed with MTBE. The combined filtrate was washed with water (500 ml, 5 vol). The aqueous layer was extracted with MTBE. The combined organic layers were washed with 5% N-acetyl-L-cysteine solution twice (300 ml, 3 vol each time) and water (300 ml, 3 vol). Then the organic layer was separated, treated with 10% active carbon, filtered through a pad of celite. The Celite layer was washed with MTBE. The combined filtrate was concentrated to 2 vol., passed through methanol twice (2 x 4 vol.) to 3 vol. and cooled to ambient temperature before use in the next step.

A solution of compound 1b was added with KHF2 (5.0 equiv.), purified water (2.6 vol.) and MeOH (1 vol.) before heating to 65°C for 1 h. The reaction mixture was diluted with MTBE (15 vol.) before cooling to 10±5°C. The resulting suspension was stirred for 1 hour before filtering. The solid was transferred to a reaction vessel, 20 vol. acetone, stirred at 25±5°C for 1 h, treated with 10% charcoal and stirred for another 1 h. The resulting reaction mixture was then filtered through a pad of celite. The filtrate was concentrated to 2 vol., passed through MTBE (3 vol.x2), concentrated to vol. 2 and diluted with MTBE (4 vol.). The suspension was stirred for 1 hour at 25±5° C. before filtration to give the desired compound 1c product as an off-white solid (11.5 g, 49.5%, HPLC 97.8% purity). ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 8.2 (s, 1H), 7.41 (m, 1H), 7.34 (m, 1H), 6.80 (m, 1H); ¹³ C NMR (100 MHz, DMSO-d6): 161.8, 138.0, 138.0, 130.8,

- 40 042196

119.4, 118.0.96.6.

Example 5 Synthesis of 6-bromo-3-chloroquinolin-4-ol (A-II).

N-Chlorosuccinimide (377 g, 1.05 equiv.) was added to a suspension of 6-bromoquinolin-4(1H)-one (Compound A-I, 600 g, 1 equiv.) in acetic acid (12 L, 20 vol.) at .T. The reaction mixture was then heated to 50° C. and stirred for 8 hours. The reaction was cooled to 20° C., filtered, washed successively with AcOH (1.8 L, 3 vol.), water (2.4 L, 4 vol.) and MTBE (1.2 L, 2 vol) and dried under vacuum on a filter to give the crude compound A-II. The crude material was stirred in MTBE (7.2 L, 12 vol) for 2 h, filtered, washed with MTBE (0.6 L, 1 vol) and dried under vacuum to give 541 g (78%) of compound A-II as an off-white solid which was 97.35% pure (HPLCAUC).

Example 6 Synthesis of 4,6-dibromo-3-chloroquinoline (A-III).

A-II A-III

Phosphorus tribromide (317 ml, 1.6 equiv.) was slowly added to a solution of compound A-II (540 g, 1 equiv.) in DMF (7 L, 13 vol.) at 0-5°C. The reaction mixture was allowed to warm to rt. and stirred for 4 hours. The reaction was cooled to 0° C. and quenched. aqueous solution of NaHCO ₃ to pH~8 (10.8 l, 20 vol.) and diluted with water (5.4 l, 10 vol.). The mixture was stirred for 2 h at RT. and the solids were filtered, washed with water (2.7 L, 5 vol.) and dried on the filter under vacuum. The wet cake was suspended in water (5.4 L, 10 vol) for 2 h and filtered, washed with water (980 ml, 2 vol) and dried on the filter under vacuum to give the crude compound A-III as a solid substances. The crude material was stirred in MTBE (2.7 L, 5 vol) for 2 h, filtered, washed with MTBE (980 ml, 2 vol) and dried under vacuum to give 434 g (65%) of compound A-III as an off-white solid that was 97.95% pure (HPLC-AUC).

Example 7 Synthesis of tert-butyl (1- (6-bromo-3-chloroquinolin-4-yl) piperidin-4-yl) carbamate (A-IV).

Diisopropylethylamine (932 ml, 4 equiv.) and 4-(N-Boc-amino)piperidine (430 g, 1.6 equiv.) were added sequentially to a solution of compound A-III (430 g, 1 equiv.) in DMSO (4 .3 l, 10 vol.) at r.t. The suspension was then heated to 140° C. and stirred for 3 hours. The reaction mixture was allowed to cool to rt, diluted with water (12.9 L, 30 vol.) and stirred for 2 hours. The resulting solids were filtered and dried on filter. The wet cake was dissolved in DCM (3 L, 7 vol) and the aqueous layer was extracted with DCM (860 ml, 2 vol). The combined organic layer was washed with water (2x2.1 L, 5 vol. each), brine (2.1 L, 5 vol.) and dried under vacuum to give the crude compound A-IV as a solid.

The crude material was stirred in MTBE (2.21 L, 5 vol) for 1 h, filtered, washed with MTBE (860 ml, 2 vol) and dried under vacuum to give 412 g (70%) of compound A-IV as an off-white solid that was 98.26% pure (HPLC-AUC).

Example 8 Alternative synthesis of tert-butyl (1-(6-bromo-3-chloroquinolin-4-yl)piperidin-4yl)carbamate (A-IV).

- 41 042196

A mixture of 6-bromo-4-chloroquinoline (25 g, 1.0 equiv.), DMF (6.0 vol.), 4-(t-butoxycarbonylamino)piperidine (2.0 equiv.), K ₂ CO3 (2, 5 equiv.) was stirred and heated to 105°C for 16 hours. The reaction was monitored by IPC-HPLC and observed 93.5% of compound 4 and 0.12% of compound 3. The reaction mixture was then cooled to 25-30°C , purified water (30 vol.) was added and stirred for 2 hours. The solid was filtered and washed with purified water. The crude solid was slurried with n-heptane (5 vol), filtered and washed with n-heptane (2 vol). The solid was dried at 55° C. to give the desired compound 4 product (35.3 g, 84% yield). ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 8.69 (d, 1H), 8.02 (d, 1H), 7.88 (m, 1H), 7.80 (m, 1H), 7.02 (d, 1H), 6.97 (d, 1H), 3.44 (m, 3H), 2.87 (m, 2H), 1.94 (d, 2H), 1.68 (m , 2H), 1.38(s, 9H).

Compound 4 (25 g, 1 equiv.), DIPA (0.078 equiv.), NCS (1.5 equiv.), toluene (10 vol.) were mixed and heated at 70°C for 4 hours. The reaction mixture was concentrated to the level 3 vol. at 45±5°C, cooled to RT, diluted with MTBE (10 vol.), Washed with purified water (10 vol.). After separation of the layer, the aqueous layer was extracted with MTBE (5.0 vol.). The combined organic layers were washed with purified water twice (2x5 vol.) and then with saline. The organic layer was dried over sodium sulfate, concentrated to 2 vol., passed through MTBE twice (2x2 vol.), then it was cooled and MTBE (1 vol.) was added before heating to 50±5°C and stirred for 1 h. The resulting suspension was cooled to 5±5° C. and stirred for 1 h. The solid was collected by filtration and washed with pre-chilled MTBE (1 vol), the above solid was taken up in MTBE (2 vol) and heated again to 55±5° C, stirred for 1 h, cooled to 5±5°C and stirred for another 1 h. The solid was collected by filtration and washed with pre-chilled MTBE (1 vol.). The collected solid was dried under reduced pressure at 45±5° C. for 8 hours to give the compound A-IV product of 98.9% HPLC purity and 66% (18 g) isolated yield. 1H NMR (400 MHz, DMSO-d ₆ ): δ 8.65 (s, 1H), 8.25 (d, 1H), 7.92 (m, 1H), 7.73 (m, 1H), 3 .76 (s, 1H), 3.51 (m, 2H), 3.37 (d, 2H), 2.14 (d, 2H), 1.69 (m, 2H), 1.46 (s, 9H).

Example 9 Synthesis of tert-butyl (1-(6-(3-cyano-2-hydroxyphenyl)-3-(3,5-difluorophenyl)quinolin-4-yl)piperidin-4-yl)carbamate (A-VI) .

Quinoline A-IV (350 g, 1 equiv.), (3-cyano-2-hydroxyphenyl)boronic acid (compound 1a) (155 g, 1.2 equiv.) and K ₂ CO ₃ (438 g, 4 equiv.) ) was loaded into a round bottom flask. 1,4-Dioxane (3.5 L, 10 vol.) and DI water (350 ml, 1 vol.) were added to the flask and the resulting reaction mixture was degassed with argon for 30 min PdCl ₂ (dppf)-CH ₂ Cl ₂ ( 32.5 g, 0.05 equiv) was added to the reaction under argon and a mixture of degas

- 42 042196 were zated for an additional 10 min. The reaction mixture was stirred at 80-85° C. and monitored by TLC and HPLC. After completion of the reaction (6 h), the mixture was allowed to cool to 25-30° C. and 3,5-difluorophenylboronic acid (346 g, 3 equiv.) was added to the reaction mixture, which was then degassed with argon for 10 min with PdCl ₂ (amphos) ( 25.9 g, 0.05 equiv) was added to the flask under argon and the reaction was degassed for an additional 10 minutes. The reaction mixture was then heated to 90-100° C. and stirred for 19 h (monitored by TLC and HPLC). 82.04% compound A-VI was observed by HPLC along with 1.95% unreacted compound AV and 0.94% other impurity at 8.2 minutes. The reaction mixture was allowed to cool to 25-30° C. and filtered through a pad of Celite and washed with ethyl acetate (1350 mL, 3 vol). The filtrate was concentrated in vacuo to ~10% solvent residue and the resulting residue was diluted with ethyl acetate (6.3 L, 18 vol.), washed with water (2x3.5 L, 10 vol. each), brine (3.5 L, 10 vol. .) and dried over anhydrous Na ₂ SO ₄ . The organic layer was concentrated to dryness in vacuo and then slurried in ethyl acetate (2.1 L, 6 vol) over 4 h at rt. (After 2.5 hours of stirring at rt, the formation of a free solid was observed). The resulting free solids were filtered, washed with ethyl acetate (700 ml, 2 vol.) and dried under vacuum until a constant weight reached 200 g (45%) of compound A-VI as an off-white solid, which was 98.4 % pure (HPLC-AUC) with approx. 3500 ppm trace palladium.

Example 10 Alternative Synthesis of tert-Butyl (1-(6-(3-cyano-2-hydroxyphenyl)-3-(3,5-difluorophenyl)quinolin-4-yl)piperidin-4-yl)carbamate (A-VI ).

CN

HE

BF ₃ K

^С1 Ρά(ΡΡή ₃ ) ₄ /Κ ₂ ΟΟ ₃

Dioxane, 85°С, 6-12 h

A-IV A-V

B(OH) ₂ NHBoc

A-VI

Compound A-IV (25.0 g, 1 equiv.), compound 1c (1.2 equiv.), K2CO3 (3.0 equiv.), 1,4-dioxane (9 vol.) and purified water (0.2 equiv.). 75 vol) was added to the reaction vessel. The mixture was degassed by bubbling N2 before adding Pd(PPh ₃ ) ₄ (0.017 equiv.). The reaction mixture was then heated to 80-85° C. for 12 hours. <1% of compound A-IV was observed at 12 hours by IPC. Then 3,5-difluorophenylboronic acid (2.0 equiv.), Pd(amphos)Cl ₂ (0.03 equiv.) was added and the reaction mixture was again degassed before heating at 90-95° C. for 6 h. 2% remaining AV compound. A sample of pure compound AV was isolated and characterized by ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 8.63 (s, 1H), 8.27 (bs, 1H), 7.94 (m, 2H), 7.53 (d, 1H, J = 7.2 Hz), 7.47 (d, 1H, J = 6.0 Hz), 6.99 (d, 1H, J = 7.6 Hz), 6, 77 (bs, 1H), 3.50 (m, 1H), 3.41 (m, 2H), 3.34 (m, 2H), 1.87 (m, 2H), 1.65 (m, 2H ), 1.39 (s, 9H).

Processing and removal of Pd.

The reaction mixture was cooled to 25-30°C, filtered through a pad of celite. The Celite layer was washed with IPc (2.0 vol.). The filtrate was combined, concentrated to 3 vol., passed through IPc (5 vol.) twice to 4 vol. The resulting solution was diluted with IPc (8 vol.) and washed with water (2x10 vol.). The organic layer was separated and washed with 1% N-acetyl-L-cysteine (2x10 vol.) before concentrating to 6 vol. The resulting suspension was stirred at reflux for 2 hours and cooled to ambient temperature. The suspension was further cooled to 10±5°C, stirred for 2 h and filtered. The filter cake was washed with 1 vol. IPAc and dried to give the desired crude product as a pale yellow solid.

The isolated crude solid compound A-VI was dissolved in 2-methyl-THF (15 vol.), SILIAMET S THIOL (0.25% w/w) was added and stirred at ambient temperature for 3 hours. The suspension was filtered through Celite layer, washed with 2-methyl-THF (2 vol.). The above process

- 43 042196 repeated again. The final filtrate was concentrated to 2 vol., passed through n-heptane twice (2x3 vol.). The resulting suspension was filtered. The solid was dried under vacuum at 45±5°C to give the crude compound A-VI as a solid (HPLC purity 96% with Pd ppm).

Example 11 Compound A-VI Further Purification.

Compound A-VI (200 g, 98.40% pure) was taken up in IPAc (1 L, 5 vol) and heated at reflux for 1 h. The mixture was then allowed to cool to rt and then cooled to 15 °C, filtered, washed with IPAc (600 ml, 3 vol.) dried to give 170 g of compound A-VI as an off-white solid which was 98.71% pure (HPLC-AUC) with approx. 50 ppm trace palladium.

Example 12 Removal of Residual Palladium from Compound A-VI.

Compound A-VI (150 g, 98.71% pure) was dissolved in THF (3.4 L, 20 vol). Si-thiol (240 g) was added and the solution was stirred overnight at rt. The mixture was filtered through a pad of Celite, washed with THF (510 mL, 3 vol) and concentrated in vacuo to give a solid. The crude solid was then diluted with IPc (1 L, 5 vol) and the slurry heated at reflux for 2 h. The mixture was then allowed to cool to rt, then cooled to 15°C, filtered, washed with IPc (510 ml 3 vol) and dried to give 150 g of compound A-VI as an off-white solid which was 100% pure (HPLC-AUC) with no detectable residual palladium.

Example 13 3-(4-(4-Aminopiperidin-1-yl)-3-(3,5-difluorophenyl)quinolin-6-yl)-2-hydroxybenzonitrile, di-HCl salt (compound A-2HC1).

Connection A-2HCI

A suspension of compound A-VI (150 g, 269.5 mmol) in 6 M HCl-IPA (1.45 L, 10 vol) was stirred at rt. within 4 h. Using TLC, the reaction was observed to be complete (a thick suspension was observed). The reaction mixture was diluted with MTBE (2.17 L, 15 vol.), stirred for 2 h and the resulting solids were filtered, washed with MTBE (290 ml, 2 vol.) and dried under vacuum to obtain 130 g (91%) of compound A -2HC1 salts as a pale yellow solid which was 100% pure (HPLC-AUC) with no detectable residual palladium. When titrated with chloride, 15.8 wt% chloride was observed.

Example 14 3-(4-(4-Aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl)-2-hydroxybenzonitrile, HCl salt (compound A-HCl).

_ A-HCI connection

A-VI

A suspension of compound A-VI (120.0 g) in IPc (5 vol) was added with 6 M HCl in isopropyl acetate (10 vol) at rt. The resulting mixture was stirred for 2 h at rt. After completion of the reaction (compound A-VI found to be <1.0% by HPLC), the thick slurry was diluted with IPc (0.6 L, 5 vol), stirred for 1 h and filtered. The filter cake was washed with IPc (240.0 ml, 2 vol) and sucked under vacuum for 2 hours to give compound A-2HC1 as a yellow solid. Yield of crude compound: 110.0 g (96%)

A solution of compound A-2HC1 (110.0 g) in purified water (18 vol.) was heated to 45±5°C before slowly adding aq. NH3 (25%, 2.4 equiv.) and stirred for 1 hour at 45±5°C. The pH value of the final reaction mixture was 5-5.5. Then purified water (550.0 ml, 5 vol.) was slowly added at the same temperature over 2 hours. The resulting suspension was cooled to rt, then to 10±5°C and stirred for 1 hour. washed with cold purified water (2 vol.) (10±5°C). The wet cake was aspirated for one hour and dried at 55±5° C. for 24 h under vacuum to give compound A-HCl as a bright yellow to golden solid (88.0 g, 83% isolated yield from 99. 4% HPLC purity).

- 44 042196

Example 15 Preparation of compound A-HCl from compound A-2HC1.

A-2HCI connection A-HCI connection

A suspension of compound A-2HC1 (118 g) in DI water (2.4 L, 20 vol) was stirred at 40-45°C. Aqueous ammonia (25%, 35.4 ml, 0.3 ml/g) was added slowly to the hot suspension to adjust pH~5-6 and stirred for 1 h. D.I.-water (590 ml, 5 vol .) was added at the same temperature over 2 hours and the suspension was allowed to cool to 25°C, then further cooled to 5-10°C using ice and water. The free solids were then filtered and washed with DI water (3x500 ml). The wet cake was dried in a vacuum oven at 55° C. for 18 hours to give a constant weight of 101 g (92%) of compound A-HCl as a pale yellow powder that was 100% pure (HPLC-AUC) with no detectable residual palladium. . When chloride was titrated, 8.1% by weight of chloride was observed.

Example 16 Alternative Preparation of Compound A-HCl from Compound A-2HC1.

A-2HCI connection A-HCI connection

A suspension of compound A-2HC1 (2.1 g) in 1:1 IPA:water (10 ml, 5 vol) was stirred at 45°C. Concentrated ammonium hydroxide (28-30%, 0.8 equiv) was added slowly and the hot slurry was stirred for 1 h. D.I. water (20 ml, 10 vol) was added at the same temperature over 2 h and the suspension was cooled to 10° C. over 2 hours. The resulting liquid slurry was then filtered and washed 15:85 with 1PA:water (2x3 vol.). The wet cake was dried under vacuum at 45° C. for 18 hours to give a constant weight of 1.42 g (78%) of compound A-HCl as a pale yellow powder. When titrated with chloride, 8.0% by weight of chloride was observed.

Example 17 Alternative preparation of compound A-HCl from compound A-2HC1 without the use of a base.

A-2HCI connection A-HCI connection

About 80 mg of compound A-2HC1 salt was dissolved in various solvents (see table below). The reaction mixtures were heated to 50°C for 30 min, cooled for 2 h to rt, kept at rt. for 1 h, then the solids were filtered and dried under vacuum at 50°C. The solvent was evaporated to give compound A-HCl and the resulting XRPD sample was examined. Each solvent system except 9:1 methyl acetate:water resulted in an XRPD pattern as in FIG. 1.

- 45 042196

di-HC1 salt, mg Solvent(s) About. XRPD sample 79.3 1RA: water (8:2) 10 Figure 1 76.7 1RA: water (7:3) 10 Figure 1 78.3 methyl acetate:water (9:1) 10 Di-HC1 79.4 methyl acetate:water (8:2) 10 Figure 1 78.7 water 6 Figure 1 79.5 water 10 Figure 1 92.7 water + 0.5 equiv. _NaHCO3 10 Figure 1

Example 18 Preparation of Compound A from Compound A-2HC1.

About 1.2 g of compound A-2HC1 was slurried in 15 sat volumes. sodium bicarbonate for 30 min. The solid was filtered and dried at 40-45° C. under vacuum to give 730 mg (77%) of Compound A as the free base.

Example 19 Preparation of Compound A-HCl from Compound A

Compound A

A-HCI compound

About 17 mg of compound A, free base was dissolved in various solvents (500 μl each, see table below) followed by the addition of 1.0 equiv. 3 M HCl/IPA solution. The solids were filtered to give compound A-HCl and the resulting XRPD sample was examined. Each solvent, with the exception of MTBE, resulted in an XRPD pattern in both FIG. 1.

Free base, mg HC1/IPA equiv. Solvent XRPD sample 18.5 1 MeOAc Figure 1 17.9 1 Acetonitrile Figure 1 17.9 1 THF Figure 1 16.1 1 EYN: water (9:1) Figure 1 17.1 1 Acetone:water (9:1) Figure 1 17.2 1 MTVE Amorphous

Example 20 X-Ray Powder Diffraction (XRPD) X-ray powder diffraction was performed using a Rigaku MiniFlex 600. Samples were obtained on zero-return Si wafers. A typical scan is 29° to 30° in 0.05° increments for five minutes at 40 kV and 15 mA. Typical XRPD parameters are outlined below.

- 46 042196

Options for reflection mode X-ray wavelength X-ray tube installation Si Καί, 1.540598 A, 40 kV, 15 mA Conditions for the gap Scan mode Fixed + variable size slot system continuously Scan angle range (°2TH) 4-30 Step size (°2TH) 0.05 Scan speed (°/min) 5

Characterization of the crystalline compound A-HCl.

The X-ray powder diffraction pattern for compound A-HCl is depicted in FIG. 1. This XRPD sample showed the crystalline form of compound A-HCl. The peaks include the peaks set forth in the following table.___________________________________________

Angle 2-Theta (°) Rel. intensity (%) 4.5 50 9.1 50 10.2 100 16.3 52 18.4 57 19.1 55 20.7 42 23.3 43 23.4 42 23.6 47 27.1 40 28.0 34

Characteristics of the crystalline compound A-2HC1.

An X-ray powder diffraction pattern for compound A-2HCl is shown in FIG. 4. This XRPD sample showed the crystalline form of compound A-2HC1. The peaks include the peaks set forth in the following table.

Angle 2-Theta (°) Rel. intensity (%) 5.4 33 7.3 100

Characterization of sample A of crystalline compound A.

The X-ray powder diffraction pattern for Sample A of Compound A is shown in FIG. 7(a). This XRPD sample showed the crystalline form of Compound A. The peaks include the peaks set forth in the following table.

Angle 2-Theta (°) Relative peak intensity, % 9.2 100 12.3 35 14.4 31 24.0 29

Characterization of sample B of crystalline compound A.

The X-ray powder diffraction pattern for sample B of compound A is shown in FIG. 7(b). This XRPD sample showed the crystalline form of compound A. The peaks include the peaks set forth in the following table.

Angle 2-Theta (°) Relative peak intensity, % 5.9 34 13.9 22 14.2 20 17.5 100 24.6 thirty

Characterization of sample C of crystalline compound A.

The X-ray powder diffraction pattern for Compound A Sample C is shown in FIG. 7(c). This XRPD sample showed the crystalline form of Compound A. The peaks include the peaks set forth in the following table.____________________________________________

Angle 2-Theta (°) Relative peak intensity, % 7.2 41 8.3 100 10.9 26 12.0 77

Example 21 Water Solubility, FaSSGF and FaSSIF.

Salt forms were suspended in water, artificial gastric juice on an empty stomach (FaSSGF) and artificial intestinal juice on an empty stomach (FaSSIF) at 37°C for 2 days. A portion of the slurry was filtered using a 0.4 μM syringe filter to recover the supernatant for solubility measurements, and the remainder was filtered to recover the remaining solid for XRPD analysis.

A calibration curve was built to measure the concentration using FLPLC. Compound A was added to a 100 ml volumetric flask and then filled to volume with FLPLC grade ethanol. In table. 3 shows the calibration concentration and the corresponding AUC. A 30 minute gradient method was used with solvent A as water with 0.1% TFA and solvent B as methanol:acetonitrile (1:1).

The calibration correlation was used to measure the solubility of the salt forms. From a clear saturated solution, 0.1 ml of liquid was transferred into a 10 ml vessel, and then the volume was filled with methanol. The solution was then immediately injected into the HPLC.

The results of solubility studies are presented in table. 4.

Table 4

Compound Water, mg/ml FaSSGF, mg/ml FaSSIF, mg/ml Compound A (free base) 0.005 2.43 0.013 Compound A-HC1 0.83 2.12 0.01 Compound A-2HC1 5.63 3.69 0.011

Solubility is a clear function of the pH value. The compounds were also found to remain as the HCl salt in FaSSGF, which stimulates gastric juices.

Example 22 Dynamic Vapor Sorption (DVS)

Dynamic Vapor Sorption (DVS) was performed using an internal DVS 1. The sample was loaded into the sample dish and removed from the microbalance. A typical sample weight for a DVS study is 25 mg. Nitrogen gas was bubbled through distilled water to maintain the required relative humidity.

- 48 042196

A typical measurement includes steps where:

1) equilibrated at 50% RH;

2) 50% to 2%. (50%, 40%, 30%, 20%, 10% and 2%)

a) a minimum retention of 5 minutes and a maximum of 60 minutes at each humidity; a criterion for passing less than 0.002% change;

3) 2% to 95% (2%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%);

a) a minimum retention of 5 minutes and a maximum of 60 minutes at each humidity; a criterion for passing less than 0.002% change;

4) 95% to 2% (95%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10%, 2%);

a) a minimum retention of 5 minutes and a maximum of 60 minutes at each humidity; a criterion for passing less than 0.002% change;

5) 2% to 50% (2%, 10%, 20%, 30%, 40%, 50%);

a) a minimum retention of 5 minutes and a maximum of 60 minutes at each humidity; the criterion for passing less than 0.002% change.

As noted below, the A-HCl compound absorbs much less moisture than the A-2HC1 compound.

Compound A-HCl.

When examining the mono-HCl salt of Compound A (compound A-HCl), a reversible uptake of water (~4.5% w/w) from 2 to 95% RH was observed; and reversible water uptake (~2.3% w/w) from 15 to 75% RH. XRPD was unchanged after DVS analysis (Fig. 9).

Compound A-2HC1.

When examining the di-HCl salt of compound A (compound A-2HC1), a reversible uptake of water (~18% w/w) from 2 to 95% RH was observed; and reversible water uptake (~9% w/w) from 15 to 75% RH. XRPD was unchanged after DVS analysis (Fig. 10).

Example 23 Thermal Studies of Compound A-HCl.

Differential scanning calorimetry (DSC) was performed using a Mettler Toledo DSC3+. An aluminum crucible with a clamped fixed cap was used. Heating rates were typically 10°C/min from 30 to 250°C.

Thermogravimetric analysis (TGA) was performed using a Mettler Toledo TGA/DSC3+. An aluminum crucible with a clamped fixed cap was used. Heating rates were typically 10°C/min from 30 to 250°C.

The protection and purge gas is nitrogen (20 - 30 ml/min and 50 - 100 ml/min). Typical parameters for DSC/TGA are listed below.

Options.

Method Linear measurement

Sample size 5-10 mg

Heating rate 10.0°C/min

Temperature range from 30 to 250°С

Compound A-HCl.

As shown in FIG. 2(a) and FIG. 2(b) compound A-HCl shows many peaks in its thermogram. These peaks were evaluated using thermal analysis and XRPD techniques. The AHCl compound was heated to various temperatures using TGA/DSC followed by cooling to room temperature and analysis using XRPD. The solid was analyzed by Karl-Fischer titration and showed 5.9% water in the sample.

The following thermal treatments were carried out:

sample 1: heated salt to 100°C and cooled to rt. followed by XRPD;

sample 2: heated salt to 150°C and cooled to rt. followed by XRPD;

sample 3: heated salt to 200°C and cooled to rt. followed by XRPD;

sample 4: heated salt to 250°C and cooled to rt. followed by XRPD;

sample 5: heated salt to 295°C and cooled to rt. followed by XRPD.

The solid was observed to be in solid form up to 200°C and melt at 220°C, at which point the compound loses its chemical and crystalline integrity. Thus peaks above 220°C are not relevant.

Since the solid contains about 5.9% water, the initial endothermic peak is due to water loss.

The DSC thermogram for compound A-2HC1 is shown in FIG. 5(a).

The TGA thermogram for compound A-2HC1 is shown in FIG. 5(b).

DSC thermogram for sample C of compound A, free base, is shown in FIG. 8(a).

TGA thermogram for sample C of compound A, free base, is shown in FIG. 8(b).

Example 24 Infrared (IR) Spectroscopy

200 mg of freshly dried potassium bromide were weighed and transferred to a mortar and ground to a fine powder.

- 49 042196 cue powder. To this was added 2.0 mg of the test compound and the solids were thoroughly mixed. A small amount of powder was turned into a thin translucent pellet. Shimadzu IR Prestige 21 was used to generate IR spectra of test compounds with 60 scans from 4000 cm ^-1 to

400 cm ^-1 . Air was used as a reference.

Characterization of the crystalline form of compound A-HCl The IR spectrum for compound A-HCl is shown in FIG. 3.

Characteristic peaks include peaks at: 2223 cm ^-1 , 1620 cm ^-1 , 1595 cm ^-1 , 1457 cm ^-1 , 1238 cm ^-1 , 1220 cm ^-1 and 1117 cm ^-1 .

Crystal Form Characterization of Compound A-2HC1 The IR spectrum for Compound A-2HC1 is shown in FIG. 6.

Characteristic peaks include peaks at: 2227 cm ^-1 , 1620 cm ^-1 , 1594 cm ^-1 , 1456 cm ^-1 , 1439 cm ^-1 , 1321 cm ^-1 and 1122 cm ^-1 .

Example 25 High Performance Liquid Chromatography (HPLC) Methods

Method A.

A Hitachi HPLC equipped with a DAD detector was used for solubility and purity studies. The HPLC column used was C18 5 µ 100A, 4.6 mm x 250 mm. Chromatographic conditions are presented in the following tables.

Mobile phase (A) 0.1% TFA in 1L Milli Q water Mobile phase (B) Acetonitrile Sample temperature 25°С Flow rate 0.8 ml/min Detector wavelength 232 nm Injection volume 5 µl Chromatography time 25 min injection delay 5 minutes

Stage Time (min) Mobile phase A (%) Mobile phase B (%) 1 OD 98 2 2 16 2 98 3 19 2 98 4 22 98 2 5 25 98 2

Method B.

A Waters Alliance HPLC (or equivalent) equipped with a DAD detector was used for purity studies. The HPLC column used was C18 5 µ 110A, 4.6 mm x 250 mm. Chromatographic conditions are presented in the following tables. ____________________________

Mobile phase (A) 0.1% OPA in 1 liter of Milli Q water Mobile phase (B) Acetonitrile Sample temperature 25°C Flow rate 1.2 ml/min Detector wavelength 232 nm/212 nm Injection volume 5 µl Chromatography time 25 min injection delay 5 minutes

- 50 042196

Stage Time (min) Mobile phase A (%) Mobile phase B (%) 1 OD 98 2 2 0.5 98 2 3 7 2 98 4 17 2 98 5 21 98 2 6 25 98 2

Example 26 2-{4-[(4aS,8aS)-Octahydro-1H-pyrido[3,4-b][1,4]oxazin-6-yl]-3-(3-fluorophenyl)quinolin-6yl} -4-methylpyridine-3-amine (1-8).

Stage 1-1. Preparation of benzyl(4aS,8aS)-6-[3-chloro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2yl)quinolin-4-yl]-octahydro-1H-pyrido [3,4-b][1,4]oxazine-1-carboxylate: To dioxane solution (3.0 ml) benzyl(4aS,8aS)-6-(6-bromo-3-chloroquinolin-4-yl)- octahydro-1 H-pyrido[3,4-b][1,4]oxazine-1-carboxylate (1.0 equiv., 0.29 mmol, 150 mg) was added 4,4,5,5-tetramethyl-2 -(4,4,5,5-tetramethyl-1,3,2-dioxaborolan2-yl)-1,3,2-dioxaborolane (2.0 equiv., 0.58 mmol, 147 mg), KOAc (3, 0 equiv., 0.87 mmol, 85 mg) and Pd(dppf)Cl ₂ (0.06 equiv., 0.02 mmol, 12 mg) under N ₂ atmosphere. The resulting mixture was heated at 90° C. for 1 hour and then the temperature was lowered to ambient temperature. The reaction solution was concentrated under vacuum to obtain 170 mg of a crude product. This material was used in the next step without further purification.

Stage 1-2. Preparation of benzyl (4aS,8aS)-6-[6-(3-amino-4-methylpyridin-2-yl)-3-chloroquinolin-4-yl]octahydro-1H-pyrido[3,4-b][1, 4]oxazine-1-carboxylate: To dioxane solution (4.0 ml) of crude benzyl(4aS,8aS)-6-[3-chloro-6-(4,4,5,5-tetramethyl-1,3,2 -dioxaborolan-2-yl)quinolin-4-yl]-octahydro-1Hpyrido[3,4-b][1,4]oxazine-1-carboxylate (1.0 equiv., 0.29 mmol, 170 mg) was added 2-bromo-4methylpyridine-3-amine (1.0 equiv., 0.29 mmol, 56 mg), Pd(Amphos)Cl ₂ (0.10 equiv., 0.029 mmol, 21 mg), K ₂ CO ₃ ( 3.0 equiv., 0.90 mmol, 125 mg) and water (0.4 ml) under N ₂ atmosphere. The resulting mixture was heated at 90° C. for 2 hours. The reaction mixture was cooled to ambient temperature and concentrated under vacuum. The resulting residue was purified by silica gel chromatography eluting with ethyl acetate/petroleum ether (3:1) to give 70 mg of the desired product as a yellow oil. MS(M+H)+=544.2.

Stage 1-3. Preparation of benzyl(4aS,8aS)-6-[6-(3-amino-4-methylnuridin-2-yl)-3-(3-fluorophenyl)quinolin-4-yl]-octahydro-1H-pyrido[3,4 -b][1,4]oxazine-1-carboxylate: To a solution of toluene (3.0 ml) benzyl(4aS,8aS)-6-[6-(3-amino-4-methylpyridin-2-yl)-3 -chloroquinolin-4-yl]-octahydro-1H-pyrido[3,4b][1,4]oxazine-1-carboxylate (1.0 equiv., 0.13 mmol, 70 mg) acid (2.0 equiv., 0.26 mmol, 36 mg), Pd(Amphos)Cl ₂ (0.10 equiv., 0.013 mmol, 9 mg), K ₂ CO ₃ (3.0 equiv., 0, 38 mmol, 53 mg) and water (0.3 ml) under N ₂ atmosphere. The resulting mixture was heated at 110° C. for 1 hour. The reaction mixture was cooled to ambient temperature and concentrated under vacuum. The resulting residue was purified by silica gel chromatography eluting with ethyl acetate/petroleum ether (1:1) to give 50 mg of the desired product as a yellow oil. MS(M+H)+ = 604.3.

Stage 1-4. Preparation of 2-{4-[(4aS,8aS)-octahydro-1H-pyrido[3,4-b][1,4]oxazin-6-yl]-3-(3fluorophenyl)quinolin-6-yl]-4 -methylpyridin-3-amine: Benzyl(4aS,8aS)-6-[6-(3-amino-4-methylpyridin-2yl)-3-(3-fluorophenyl)quinolin-4-yl]-octahydro-1H- pyrido[3,4-b][1,4]oxazine-1-carboxylate (1.0 equiv., 0.08 mmol, 50 mg) was combined with trifluoroacetic acid (2.0 ml) and the resulting mixture was heated at 80° C for 2 hours. The reaction solution was cooled to ambient temperature and concentrated under vacuum. The resulting residue was purified by prep-HPLC under the following conditions (prep HPLC-007): column, SunFire Prep C18 OBD column, 19x150 mm, 5 µm, 10 nm; mobile phase, water (0.1% TFA) and ACN (2.0% ACN to 20.0% in 6 min, hold at 95% for 1 min, drop to 2.0% in 1 min, hold at 2 .0% within 1 min); detector, UV 220 nm. This resulted in 11.4 mg (27%) of the formic acid salt 2-{4-[(4aS,8aS)-octahydro-1H-pyrido[3,4-b][1,4]oxazin-6-yl]- 3-(3fluorophenyl)quinolin-6-yl}-4-methylpyridin-3-amine as a yellow solid. MS(M+H)+=470.2.

The following compounds were prepared similarly to Example 25 with appropriate substitution of reagents and

- 51 042196 substrates at various stages and additional functional group modifications by well known chemistry with appropriate reagents as needed. Various salts such as HCl or formic acid can be prepared by conventional methods.

connection no. MS (M+H) ⁺ 1-1 517.2 1-2 527.3 1-3 547.1 1-4 495.2 1-5 515.2 1-6 517.2 1-7 481.2 1-9 471.2 1-10 471.2

Example A-1. Parenteral pharmaceutical composition.

To obtain a parenteral pharmaceutical composition suitable for administration by injection (subcutaneous, intravenous), 1-100 mg of a water-soluble salt of the compound disclosed in the present invention or a pharmaceutically acceptable salt or solvate thereof was dissolved in sterile water, and then mixed with 10 ml of 0 .9% sterile saline. A suitable buffer is optionally added as well as an optional acid or base to adjust the pH. The mixture was formulated into a unit dosage form suitable for administration by injection.

Example A-2. oral solution.

To prepare a pharmaceutical composition for oral delivery, a sufficient amount of a compound disclosed in the present invention, or a pharmaceutically acceptable salt thereof, was added to water (with optional solubilizer(s), optional buffer(s), and taste masking excipients) to give 20 mg/ml solution.

Example A-3. Oral tablet.

A tablet is prepared by mixing 20-50% by weight of a compound disclosed in the present invention or a pharmaceutically acceptable salt thereof, 20-50% by weight of microcrystalline cellulose, 110% by weight of low-substituted hydroxypropyl cellulose and 1-10% by weight of magnesium stearate or other suitable excipients. Tablets were obtained by direct compression. The total weight of the compressed tablets was maintained at 100-500 mg.

Example A-4. oral capsule.

To prepare a pharmaceutical composition for oral delivery, 10-500 mg of a compound disclosed in the present invention, or a pharmaceutically acceptable salt thereof, was mixed with starch or other suitable powder mixtures. The mixture was included in a dosage unit for oral delivery, such as a hard gelatin capsule, which is suitable for oral administration.

In another embodiment, 10-500 mg of a compound disclosed in the present invention, or a pharmaceutically acceptable salt thereof, is placed in a size 4 capsule or a size 1 capsule (hypromellose or hard gelatin) and the capsule is capped.

Example A-5. Gel composition for topical application.

To obtain a pharmaceutical gel composition for topical application, the compound disclosed in the present invention, or a pharmaceutically acceptable salt thereof, was mixed with hydroxypropyl cellulose, propylene glycol, isopropyl myristate and purified with USP alcohol. The resulting gel mixture is then included in containers, such as tubes, which are suitable for topical application.

Example B-1. SSTR analyses.

Functional analysis for SSTR2 agonists.

Overview: All five SSTR subtypes are G-protein coupled receptors (GPCRs) that result in a decrease in intracellular cyclic AMP (cAMP) when activated by an agonist. Thus, the measurement of intracellular cAMP levels can be used to determine whether the compounds of the present invention are SSTR subtype agonists (John Kelly, Troy Stevens, W. Joseph Thompson, and Roland Seifert, Current Protocols in Pharmacology, 2005, 2,2,1 -2.2). Analysis of intracellular cAMP in association with SSTR2 is described below. Human SSTR assays 1, 3, 4, and 5 follow the same protocols as SSTR2.

cAMP analysis protocol.

Four days prior to analysis of 5000 Chinese hamster ovary cells (CHO-K1, ATCC #CCL-61),

- 52 042196 stably expressing human SSTR2 were placed in each well of a 96-well tissue culture plate treated with Ham's F12 Growth Medium (ThermoFisher No. 10-080-CM) supplemented with 10% donor bovine serum (Gemini Bio-Products No. 100 -506), 100 U/ml penicillin; 100 µg/ml streptomycin; 2 mM L-glutamine (Gemini Bio-Products no. 400-110) and 0.2 mg/ml hygromycin B (GoldBio no. 31282-04-9). Cells were cultured at 37°C, 5% CO2 and 95% humidity. On the day of analysis, medium was aspirated and cells were treated with 50 µl of 1.6 µM NKH477 (Sigma No. N3290) together with various dilutions of compounds of the present invention in assay buffer [1x Hank's balanced salt solution (ThermoFisher No. SH3058802), 0.5 mM HEPES pH 7.4, 0.1% bovine serum albumin, 0.2 mM 3-isobutyl-1-methixanthine (IBMX, VWR No. 200002-790)]. Cells were incubated for 20 min at 37°C (final concentration of the compounds of the present invention is typically 0-10,000 nM). Cells were treated with 50 μl lysis buffer (FIRTF cAMP kit, Cisbio). The lysate was transferred to 384-well plates, the cAMP detection kit and imaging antibodies were added and incubated for 1-24 hours at room temperature. The time resolved fluorescent signal was read using a Tecan M1000Pro multiplate reader. Intracellular cAMP concentrations were calculated from regression with a standard curve and plotted against the concentration of compounds of the present invention and EC ₅₀ compounds were calculated using standard methods. All data manipulations were performed in GraphPad Prism v6.

Exemplary biological activity of the compounds was demonstrated in the following table by assessing the inhibition of cAMP activities by human SST2R, where A means <10 nM; B means >10 nM and <100 nM; C means >100 nM and <1000 nM; D means >1000 nM.

The examples and embodiments described herein are for illustrative purposes only, and various modifications or changes suggested by those skilled in the art are within the spirit and scope of the present invention and the scope of the appended claims.

Claims (27)

1. 3-[4-(4-Aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride, or a hydrate thereof, of the following structure: (2) bromination of compound A-II with a suitable brominating agent in a suitable solvent to obtain compound A-III - 56 042196 and (3) subjecting 4-(N-Boc amino)piperidine to a coupling reaction with Compound A-III in the presence of a suitable base and in a suitable solvent to give Compound A-IV; or (i) subjecting 4-(N-Boc amino)piperidine to a coupling reaction with 6-bromo-4-chloro-quinoline in the presence of a suitable base and in a suitable solvent to give compound 4; NHBoc Compound 4; and (ii) chlorinating compound 4 with a suitable chlorinating agent in a suitable solvent to give compound A-IV. 2. Crystalline 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or its hydrate. 3. Crystalline 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or its hydrate according to claim 2, characterized by powder diffraction radiograph (XRPD) with peaks at 4.5° 2-Theta, 9.1° 2-Theta, 10.2° 2-Theta, 16.3° 2-Theta, 18.4° 2-Theta and 19.1 ° 2-Theta; X-ray powder diffraction (XRPD), which is shown in FIG. 1; - 53 042196 differential scanning calorimetry (DSC) thermogram with an endotherm starting at about 207° C. and peaking at about 220° C.; differential scanning calorimetry (DSC) thermogram, which is shown in FIG. 2(a); infrared spectrum (IR) with peaks at 2223 cm ^-1 , 1620 cm ^-1 , 1595 cm ^-1 , 1457 cm ^-1 , 1238 cm ^-1 , 1220 cm ^-1 and 1117 cm ^-1 ; infrared (IR), as shown in FIG. 3; unchanged XRPD when heated to about 200°C, when exposed to more than 90% relative humidity for about 24 hours, or when exposed to about 75% relative humidity (RH) and 40°C for one week, or combinations thereof; or their combinations. 4. Crystalline 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or its hydrate according to claim 2, characterized by powder diffraction radiograph (XRPD) with peaks at 4.5° 2-Theta, 9.1° 2-Theta, 10.2° 2-Theta, 16.3° 2-Theta, 18.4° 2-Theta and 19.1° 2 -Theta. 5. Crystalline 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or its hydrate according to claim 2, characterized by powder diffraction radiograph (XRPD), which is shown in Fig. 1. 6. Crystalline 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or a hydrate thereof according to any one of claims 2-5 , characterized by a differential scanning calorimetry (DSC) thermogram with an endotherm starting at about 207°C and a peak at about 220°C. 7. Crystalline 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or a hydrate thereof according to any one of claims 2-5 , characterized by a differential scanning calorimetry (DSC) thermogram as shown in FIG. 2(a). 8. Crystalline 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or a hydrate thereof according to any one of claims 2-7 , characterized by an unchanged XRPD when heated to about 200°C. 9. Crystalline 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or a hydrate thereof according to any one of claims 2-8 , characterized by unchanged XRPD when exposed to more than 90% relative humidity for 24 hours and when exposed to about 75% relative humidity (RH) and 40°C for one week. 10. Crystalline 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or a hydrate thereof according to any one of claims 2-9 , characterized by an infrared spectrum (IR) with characteristic peaks at 2223 cm ^-1 , 1620 cm ^-1 , 1595 cm ^-1 , 1457 cm ^-1 , 1238 cm ^-1 , 1220 cm ^-1 and 1117 cm ^-1 . 11. Crystalline 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or a hydrate thereof according to any one of claims 2-9 , characterized by an infrared spectrum (IR), as shown in FIG. 3. 12. Pharmaceutical composition containing crystalline 3-[4-(4-aminopiperidin-1-yl)-3(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or its hydrate according to any one of paragraphs. 2-11 and at least one pharmaceutically acceptable excipient. 13. The pharmaceutical composition according to claim 12, wherein the crystalline 3-[4-(4-aminopiperidin-1-yl)-3(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or its hydrate is is crystalline 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2hydroxybenzonitrile monohydrochloride or a hydrate thereof according to claim 3. 14. The pharmaceutical composition according to claim 12 or 13, wherein the pharmaceutical composition is in the form of a solid pharmaceutical composition. 15. The pharmaceutical composition according to any one of claims 12-14, wherein the pharmaceutical composition is in the form of a tablet, pill or capsule. 16. A process for preparing crystalline 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or a hydrate thereof, the process comprising the steps of which (i) are converted into a suspension of 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile dihydrochloride in 5 volumes of isopropanol:water (1:1); (ii) heating the slurry (i) to about 45°C; (iii) adding from about 0.5 to about 1.2 equivalents of ammonium hydroxide solution, sodium bicarbonate solution, or sodium hydroxide solution to the heated slurry of step (ii) to obtain a pH value of about 4.0-6.0; (iv) adding water over about 2 hours to the mixture of step (iii); and (v) filtering the suspension of step (iv) to obtain 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or a hydrate thereof. 17. The method according to claim 16, characterized in that the ammonium hydroxide solution is used in (iii). - 54 042196 18. A process for preparing crystalline 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or a hydrate thereof, the process comprising the steps of which (i) add a suitable solvent to 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)quinolin-6-yl]-2-hydroxybenzonitrile; (ii) add about 1 equivalent of hydrochloric acid to the mixture of solvent and 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile (i); and (iii) filtering the solids from step (ii) to give 3-[4-(4aminopiperidin-1-yl)-3-(3,5-difluorophenyl)quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or its hydrate. 19. The method of claim 18, wherein the suitable solvent in (i) is methanol, ethanol, isopropyl alcohol, acetone, methyl acetate, ethyl acetate, tetrahydrofuran, tetrahydropyran, water, or combinations thereof. 20. A process for preparing crystalline 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or a hydrate thereof, the process comprising the steps of which (i) stir 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile dihydrochloride in from about 20 volumes to about 50 volumes water; and (ii) filtering the solids of step (i) to give 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride or its hydrate . 21. Method for producing 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride NC which includes the steps in which (1) process the compound A-VI Compound A-VI with hydrochloric acid in a suitable solvent to give 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile dihydrochloride; and (2) treat 3-[4-(4-aminopiperidin-1-yl)-3-(3,5-difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile dihydrochloride with aqueous ammonia to give 3-[4- (4-aminopiperidin-1-yl)-3-(3,5difluorophenyl)-quinolin-6-yl]-2-hydroxybenzonitrile monohydrochloride. 22. The method of claim 21, wherein the suitable solvent is isopropyl alcohol, ethyl acetate or isopropyl acetate. 23. The method according to claim 21 or 22, characterized in that the compound A-VI is obtained by (1) carrying out the reaction of the compound A-IV with connection 1 - 55 042196 OH to CN Compound 1 where B in compound 1 is boronic acid, boronic acid ester or trifluoroborate; in the presence of an addition catalyst, a suitable base, and in a suitable solvent to give compound A-V and (2) reacting Compound A-V with 3,5-difluorophenylboronic acid B(OH) ₂ F'^^F in the presence of an addition catalyst, a suitable base and in a suitable solvent to give compound A-VI. 24. The method according to claim 23, characterized in that B in compound 1 is boronic acid or trifluoroborate; the addition catalyst of step (1) is a palladium catalyst; a suitable base of step (1) is triethylamine, diisopropylethylamine, 1,2,2,6,6-pentamethylpiperidine, tributylamine, sodium bicarbonate, Na ₂ CO ₃ , K ₂ CO ₃ , Cs ₂ CO ₃ , NaOAc, KOAc, Ba( OH) ₂ , Na ₃ PO ₄ or K ₃ PO ₄ ; a suitable solvent of step (1) is acetonitrile, dimethylformamide, ethanol, tetrahydrofuran, isopropyl alcohol, 1,4-dioxane, water, or combinations thereof; the addition catalyst of step (2) is a palladium catalyst; a suitable base of step (2) is triethylamine, diisopropylethylamine, 1,2,2,6,6-pentamethylpiperidine, tributylamine, sodium bicarbonate, Na ₂ CO ₃ , K2CO3, Cs ₂ CO ₃ , NaOAc, KOAc, Ba(OH) ₂ , Na ₃ PO ₄ or K ₃ PO ₄ ; and a suitable solvent of step (2) is acetonitrile, dimethylformamide, ethanol, tetrahydrofuran, isopropyl alcohol, 1,4-dioxane, water, or combinations thereof. 25. The method according to claim 23 or 24, characterized in that the compound A-IV is obtained by (1) chlorination of the compound A-I ABOUT H Compound A-I with a suitable chlorinating agent in a suitable solvent to obtain compound A-II HE Connection A-P; 26. The method according to claim 25, characterized in that the chlorinating agent of step (1) is N-chlorosuccinimide, trichloroisocyanuric acid, sulfuryl chloride, chlorine, sodium hypochlorite, calcium hypochlorite, hypochlorous acid, or 2,3,4,5,6, 6-hexachloro-2,4-cyclohexadien-1-one; a suitable solvent of step (1) is acetic acid, water, ethanol, methanol, toluene, dichloromethane, tetrahydrofuran, dioxane, or N-N-dimethylformamide; the brominating agent of step (2) is phosphorus tribromide, phosphorus oxybromide, hydrobromic acid, bromine, or dibromotriphenylphosphorane; a suitable solvent of step (2) is acetonitrile, water, ethanol, isopropanol, dichloromethane, toluene, N,N-dimethylformamide, acetic acid or acetone; a suitable base of step (3) is triethylamine, diisopropylethylamine, 1,8diazabicycloundec-7-ene, 1,2,2,6,6-pentamethylpiperidine, tributylamine, sodium bicarbonate, Na ₂ CO ₃ , K2CO3 or CS2CO3; and a suitable solvent of step (3) is N,N-dimethylformamide, N,Ndimethylacetamide, dimethylsulfoxide, dichloromethane, chloroform, carbon tetrachloride, dioxane, tetrahydrofuran, toluene, acetonitrile, ethanol or isopropanol. 27. The method according to claim 25, characterized in that the chlorinating agent of step (ii) is N-chlorosuccinimide, trichloroisocyanuric acid, sulfuryl chloride, chlorine, sodium hypochlorite, calcium hypochlorite, hypochlorous acid, or 2,3,4,5,6, 6-hexachloro-2,4-cyclohexadien-1-one; and a suitable solvent of step (ii) is acetic acid, water, ethanol, methanol, toluene, dichloromethane, tetrahydrofuran, dioxane or N,N-dimethylformamide.