CN111094272B

CN111094272B - Pharmaceutically acceptable salt and crystal form of OTR inhibitor and preparation method

Info

Publication number: CN111094272B
Application number: CN201980004485.2A
Authority: CN
Inventors: 杨俊然; 杜振兴; 王捷
Original assignee: Jiangsu Hengrui Medicine Co Ltd; Shanghai Hengrui Pharmaceutical Co Ltd
Current assignee: Jiangsu Hengrui Medicine Co Ltd; Shanghai Hengrui Pharmaceutical Co Ltd
Priority date: 2018-06-20
Filing date: 2019-06-19
Publication date: 2022-05-27
Anticipated expiration: 2039-06-19
Also published as: WO2019242642A1; CN111094272A; TW202016091A

Abstract

Provides a medicinal salt and a crystal form of an OTR inhibitor and a preparation method thereof. In particular to a medicinal salt and a crystal form of 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine serving as an OTR inhibitor and a preparation method thereof. The medicinal salt improves the free alkali dissolution degree and the physical or chemical stability of the OTR inhibitor, and has important significance for developing medicaments which are suitable for industrial production and have good biological activity.

Description

Pharmaceutically acceptable salt and crystal form of OTR inhibitor and preparation method

Technical Field

The disclosure provides a compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine pharmaceutically acceptable salt, a crystal form and a preparation method thereof.

Background

PCT/CN2017/117421 (application date 2017.12.20) describes a compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine, which shows a high-selectivity OTR inhibition effect as a free base, has good brain permeability and can effectively block the downstream function of an oxytocin receptor mediated by oxytocin.

Nearly half of the drug molecules exist in the form of salts, and the salt formation can improve some undesirable physicochemical or biological properties of the drug. Compared with 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine, the development of the salt with more excellent properties in the aspects of physical and chemical properties or pharmaceutical properties is of great significance.

Meanwhile, in view of the importance of the crystal form and the stability of the solid medicine to the clinical treatment, the polymorphism of the medicinal salt of the compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine is deeply researched, and the method has important significance for developing medicines which are suitable for industrial production and have good biological activity.

Disclosure of Invention

The present disclosure provides pharmaceutically acceptable salts of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine, wherein the pharmaceutically acceptable salts are selected from the hydrochloride, sulfate, mesylate, phosphate, citrate, benzoate, or fumarate salts.

Preferably, the chemical ratio of the compound to the acid molecule is about 1:2 to about 2:1, and may be about 1:2, 1:1, or 2: 1.

In an alternative embodiment, the chemical ratio of the compound to hydrogen chloride is about 1: 1.

In alternative embodiments, the chemical ratio of the compound to sulfuric acid is about 1:1 or 2: 1.

In alternative embodiments, the chemical ratio of the compound to phosphoric acid is about 1:1, 2: 1.

In an alternative embodiment, the compound is present in a ratio of about 1:1 stoichiometrically to the methanesulfonic acid.

Also provided in the present disclosure is a process for preparing the aforementioned pharmaceutically acceptable salts, comprising: salifying the compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine with acid.

The solvent used for salt formation in the present disclosure is selected from at least one of water, alcohols, halogenated hydrocarbons, ethers, nitriles, alcohols, esters or ethers, preferably from at least one of isopropanol, acetone, methyl tert-butyl ether, acetonitrile, ethanol, ethyl acetate, water.

In an alternative embodiment, the process for preparing the aforementioned pharmaceutically acceptable salts further comprises the steps of evaporating the solvent or stirring for crystallization, and filtering.

Also provided in the present disclosure is a pharmaceutical composition comprising a pharmaceutically acceptable salt of the aforementioned compound and a pharmaceutically acceptable adjuvant, optionally selected from at least one of a pharmaceutically acceptable carrier, diluent or excipient.

The present disclosure also provides the use of the aforementioned pharmaceutically acceptable salts in the manufacture of a medicament for the treatment or prevention of a disease or condition known or shown to produce a beneficial effect from inhibition of oxytocin, selected from sexual dysfunction, hypoactive sexual desire disorder, sexual arousal disorder, orgasmic disorder, dyspareunia disorder, premature ejaculation, pre-partum labor, complications of labor, appetite and eating disorders, benign prostatic hyperplasia, premature labor, dysmenorrhea, congestive heart failure, arterial hypertension, liver cirrhosis, renal hypertension, ocular hypertension, obsessive-compulsive disorders and neuropsychiatric disorders, preferably selected from sexual dysfunction, sexual arousal disorder, orgasmic disorder, dyspareunia disorder and premature ejaculation.

The present disclosure also provides the use of the aforementioned pharmaceutically acceptable salts in the manufacture of a medicament for antagonizing oxytocin.

The disclosure provides a crystal form A of a compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine hydrochloride, and an X-ray powder diffraction pattern expressed by a diffraction angle 2 theta angle has characteristic peaks at 6.92, 12.54, 15.23, 16.32, 22.70, 27.44 and 28.10.

In an alternative embodiment, said form a, having an X-ray powder diffraction pattern expressed in diffraction angle 2 Θ angles, has characteristic peaks at 6.92, 12.54, 15.23, 16.32, 18.89, 19.52, 20.90, 22.70, 27.44, 28.10.

In some embodiments, the form a, an X-ray powder diffraction pattern expressed in diffraction angle 2 Θ angles, has characteristic peaks at 6.92, 12.54, 15.23, 16.32, 16.63, 18.15, 18.89, 19.52, 20.90, 22.70, 24.93, 25.80, 27.44, 28.10.

In other embodiments, the form a has an X-ray powder diffraction pattern, expressed in terms of diffraction angle 2 Θ angles, as shown in figure 1.

A method for preparing a crystal form a compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine hydrochloride, comprising:

(a) adding the compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine into a solvent (I), stirring for dissolving or heating for dissolving,

(b) dripping hydrochloric acid, stirring and crystallizing;

in the method, the volume (ml) of the solvent (I) is 1 to 50 times of the weight (g) of the compound, and may be 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49 and 50 times; the solvent (I) is preferably selected from isopropanol/water, ethyl acetate.

The disclosure provides a B crystal form of a compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine hydrochloride, and an X-ray powder diffraction pattern expressed by a diffraction angle 2 theta angle has characteristic peaks at 14.62, 15.65, 19.21, 23.66, 24.15, 25.92 and 27.19.

In an alternative embodiment, said form B, having an X-ray powder diffraction pattern expressed in terms of diffraction angle 2 Θ, has characteristic peaks at 11.76, 14.62, 15.65, 19.21, 23.66, 24.15, 25.92, 27.19, 28.01, 29.54.

In some embodiments, the form B has an X-ray powder diffraction pattern, expressed in terms of diffraction angle 2 Θ, having characteristic peaks at 11.76, 14.62, 15.65, 16.43, 18.70, 19.21, 23.66, 24.15, 24.56, 25.60, 25.92, 27.19, 28.01, 29.54.

In other embodiments, form B has an X-ray powder diffraction pattern, expressed in terms of diffraction angle 2 θ, as shown in figure 2.

A method for preparing a compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine hydrochloride crystal form B, comprising:

(a) adding the compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidine-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine into a solvent (II), stirring for dissolving or heating for dissolving,

(b) dropwise adding hydrochloric acid, stirring for crystallization, and pulping;

in the method, the volume (ml) of the solvent (II) is 1 to 50 times of the weight (g) of the compound, and may be 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 times; the solvent (II) is preferably selected from ethanol, acetone, acetonitrile.

The disclosure also provides a C crystal form of the compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine hydrochloride, and an X-ray powder diffraction pattern expressed by a diffraction angle 2 theta angle has characteristic peaks at 6.94, 21.51, 22.71, 24.99, 25.80, 27.45 and 28.14.

In an alternative embodiment, said form C, having an X-ray powder diffraction pattern expressed in terms of diffraction angle 2 Θ, has characteristic peaks at 6.94, 12.54, 16.30, 20.89, 21.51, 22.71, 24.99, 25.80, 27.45, 28.14.

In some embodiments, the crystalline form C has an X-ray powder diffraction pattern, expressed in terms of diffraction angle 2 Θ, having characteristic peaks at 6.94, 12.54, 16.30, 18.15, 18.88, 19.50, 20.89, 21.51, 22.71, 24.78, 24.99, 25.80, 27.45, 28.14.

In other embodiments, the form C has an X-ray powder diffraction pattern, expressed in terms of diffraction angle 2 θ, as shown in figure 4.

A method of preparing the form C of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine hydrochloride comprising:

(a) adding the compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine into a solvent (III), stirring for dissolving or heating for dissolving,

(b) dripping hydrochloric acid, stirring and crystallizing;

in the method, the volume (ml) of the solvent (III) is 1-50 times of the weight (g) of the compound, and can be 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49 and 50 times, and the solvent (III) is preferably methyl tert-butyl ether.

The disclosure also provides a crystal form A of the compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine sulfate, and an X-ray powder diffraction pattern expressed by a diffraction angle 2 theta angle, wherein the crystal form A has characteristic peaks at 12.66, 18.50, 19.90, 21.64, 23.61, 24.22 and 26.34.

In an alternative embodiment, the form a of the sulfate salt has an X-ray powder diffraction pattern, expressed in terms of diffraction angle 2 Θ, having characteristic peaks at 12.66, 14.65, 18.50, 19.90, 21.64, 22.05, 23.61, 24.22, 24.75, 26.34.

In some embodiments, form a of the sulfate salt has an X-ray powder diffraction pattern expressed in diffraction angle 2 Θ angles with characteristic peaks at 7.28, 12.66, 14.04, 14.65, 17.60, 18.50, 19.90, 21.64, 22.05, 23.61, 24.22, 24.75, 26.34, 26.70.

In other embodiments, the form a of the sulfate salt has an X-ray powder diffraction pattern, expressed in terms of diffraction angle 2 Θ angles, as shown in fig. 7.

A method of preparing the crystal form a of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine sulfate comprising:

(a) adding the compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine into a solvent (IV), stirring for dissolving or heating for dissolving

(b) Dropwise adding sulfuric acid, stirring and crystallizing;

in this method, the volume (ml) of the solvent (IV) is 1 to 40 times of the weight (g) of the compound, and may be 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 times; the solvent (IV) is preferably selected from acetonitrile, ethanol, acetone, ethyl acetate.

The disclosure also provides a B crystal form of the compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine sulfate, and an X-ray powder diffraction pattern expressed by a diffraction angle 2 theta angle has characteristic peaks at 11.87, 13.22, 14.62, 15.29, 18.49, 22.66 and 23.61.

In an alternative embodiment, the form B of the sulfate salt of the compound has an X-ray powder diffraction pattern, expressed in terms of diffraction angle 2 Θ, having characteristic peaks at 11.87, 13.22, 14.62, 15.29, 18.49, 19.89, 21.65, 22.66, 23.61, 24.18.

In some embodiments, the form B of the sulfate salt of the compound has an X-ray powder diffraction pattern, expressed in terms of diffraction angle 2 Θ angles, having characteristic peaks at 7.78, 11.87, 12.66, 13.22, 14.62, 15.29, 18.49, 19.89, 21.65, 22.66, 23.61, 24.18, 24.62, 26.36.

In other embodiments, the form B of the sulfate salt of the compound has an X-ray powder diffraction pattern, expressed in degrees of diffraction angle 2 Θ, as shown in fig. 9.

A method of preparing form B of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine sulfate comprising:

(a) adding the compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine into a solvent (V), stirring for dissolving or heating for dissolving

(b) Dropwise adding sulfuric acid, stirring and crystallizing;

in the method, the volume (ml) of the solvent (V) is 1 to 40 times of the weight (g) of the compound, and may be 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39 and 40 times; the solvent (V) is preferably methyl tert-butyl ether.

The disclosure also provides a crystal form A of the compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine mesylate, and an X-ray powder diffraction pattern expressed by a diffraction angle 2 theta angle, wherein the crystal form A has characteristic peaks at 6.66, 16.81, 16.90, 18.92, 19.19, 22.92 and 24.88.

In an alternative embodiment, form a of the mesylate salt of the compound has an X-ray powder diffraction pattern, expressed in degrees of diffraction angle 2 Θ, having characteristic peaks at 6.66, 11.00, 16.81, 16.90, 18.92, 19.19, 21.06, 22.92, 23.96, 24.88.

In some embodiments, form a of the mesylate salt of the compound has an X-ray powder diffraction pattern, expressed in degrees of diffraction angle 2 Θ, with characteristic peaks at 6.66, 11.00, 14.87, 16.81, 16.90, 18.92, 19.19, 20.27, 21.06, 22.92, 23.96, 24.88, 27.85, 29.50.

In other embodiments, the form a of the mesylate salt of the compound has an X-ray powder diffraction pattern, expressed in degrees of diffraction angle 2 Θ, as shown in figure 14.

A method of preparing the crystalline form a of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine mesylate, comprising:

(a) adding the compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine into a solvent (VI), stirring for dissolving or heating for dissolving

(b) Dripping methanesulfonic acid, stirring and crystallizing;

in the method, the volume (ml) of the solvent (VI) is 1 to 40 times of the weight (g) of the compound, and may be 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49 and 50 times; the solvent (VI) is preferably selected from ethyl acetate, methyl tert-butyl ether.

Also provided in the present disclosure is a pharmaceutical composition comprising the aforementioned crystalline forms of a pharmaceutically acceptable salt and a pharmaceutically acceptable adjuvant, optionally from a pharmaceutically acceptable carrier, diluent or excipient.

The disclosure also provides a pharmaceutical composition prepared from the crystal form.

Also provided in the present disclosure is the use of a crystalline form of the aforementioned pharmaceutically acceptable salt for the manufacture of a medicament for the treatment or prevention of a disease or condition known or shown to produce a beneficial effect from the inhibition of oxytocin, selected from the group consisting of sexual dysfunction, hypoactive sexual desire disorder, sexual arousal disorder, orgasm disorder, dyspareunia disorder, premature ejaculation, pre-partum delivery, complications of labor, appetite and eating disorders, benign prostatic hyperplasia, premature labor, dysmenorrhea, congestive heart failure, arterial hypertension, liver cirrhosis, renal hypertension, ocular hypertension, obsessive-compulsive disorders and neuropsychiatric disorders, preferably selected from the group consisting of sexual dysfunction, sexual arousal disorder, orgasm disorder, dyspareunia disorder and premature ejaculation.

The present disclosure also provides the use of a crystalline form of the aforementioned pharmaceutically acceptable salt for the preparation of a medicament for antagonizing oxytocin.

According to the guiding principle of hygroscopicity of 9103 medicaments in the fourth part of 2015 edition of Chinese pharmacopoeia, namely the description of characteristics of hygroscopicity and the definition of hygroscopicity and weight increment,

deliquescence: absorbing sufficient water to form a liquid;

has the characteristics of moisture absorption: the moisture-inducing weight is not less than 15%;

moisture absorption: the moisture-inducing weight is less than 15% but not less than 2%;

slightly hygroscopic: the moisture-inducing weight is less than 2% but not less than 0.2%;

no or almost no hygroscopicity: the moisture-drawing weight gain is less than 0.2 percent.

The crystal form a of 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine hydrochloride in the present disclosure induces a moisture gain of 0.769% and a slight moisture attraction under the conditions of 20.0% RH to 80.0% RH.

The crystalline form B of 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine hydrochloride described in this disclosure is 20.0% RH-80.0% RH, has a moisture pick-up of 0.0641%, and has no or little moisture pick-up.

The crystalline form a of 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine mesylate described in this disclosure is 20.0% RH-80.0% RH, is increased in moisture wicking by 0.1061%, and is free or nearly free of moisture wicking.

The "X-ray powder diffraction pattern" described in this disclosure is measured using Cu-ka radiation.

The "X-ray powder diffraction pattern or XRPD" referred to in this disclosure means the pattern obtained by dividing the X-ray powder according to bragg formula 2d sin θ ═ n λ (where λ is the wavelength of the X-ray,

the order n of diffraction is any positive integer, and generally takes the first order diffraction peak, n is 1, when the X-ray is at grazing angle theta (incidence)Complementary angles of angles, also called bragg angles) are incident on an atomic plane with d-lattice plane spacing of a crystal or a part of a crystal sample, the bragg equation is satisfied, and the set of X-ray powder diffraction patterns is obtained.

The "2 θ or 2 θ angle" referred to in this disclosure refers to the diffraction angle, θ being the bragg angle in degrees or degrees; the error range of each characteristic peak 2 θ is ± 0.30, and may be-0.30, -0.29, -0.28, -0.27, -0.26, -0.25, -0.24, -0.23, -0.22, -0.21, -0.20, -0.19, -0.18, -0.17, -0.16, -0.15, -0.14, -0.13, -0.12, -0.11, -0.10, -0.09, -0.08, -0.07, -0.06, -0.05, -0.04, -0.03, -0.02, -0.01, 0.00, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.21, 0.20, 0.22, 0.23, 0.26, 0.20, preferably ± 0.20.

The term "interplanar spacing or interplanar spacing (d value)" as used in this disclosure means that the spatial lattice selects 3 non-parallel unit vectors a, b, c connecting two adjacent lattice points, which divide the lattice into juxtaposed parallelepiped units, called interplanar spacing. The space lattice is divided according to the determined connecting lines of the parallelepiped units to obtain a set of linear grids called space grids or lattices. The lattice and the crystal lattice respectively reflect the periodicity of the crystal structure by using geometrical points and lines, and the surface spacing (namely the distance between two adjacent parallel crystal surfaces) of different crystal surfaces is different; has a unit of

Or angstroms.

In the present disclosure, "differential scanning calorimetry or DSC" refers to measuring the temperature difference and heat flow difference between a sample and a reference substance during the temperature rise or constant temperature process of the sample to characterize all the physical changes and chemical changes related to the thermal effect, and obtain the phase change information of the sample.

The drying temperature in the present disclosure is generally 25 ℃ to 100 ℃, preferably 40 ℃ to 70 ℃, and the drying may be performed under normal pressure or under reduced pressure. Preferably, the drying is carried out under reduced pressure.

The chemical or biological agents used in the present disclosure may be purchased commercially.

Test conditions for the instruments used in the experiments of this disclosure:

1. differential Scanning Calorimeter (DSC)

The instrument model is as follows: mettler Toledo DSC 3⁺STAR^e System

And (3) purging gas: nitrogen gas

Rate of temperature rise: 10.0 ℃/min

Temperature range: 25-250 deg.C

2. X-ray Powder Diffraction Spectroscopy (XRPD)

(1) The instrument model is as follows: bruker D8 Discover A25X-ray powder diffractometer

Ray: monochromatic Cu-Ka ray (lambda as 1.5406)

The scanning mode is as follows: θ/2 θ, scan range: 10-48 degree

Voltage: 40KV, current: 40mA

3. Thermogravimetric analyzer (TGA)

The instrument model is as follows: mettler Toledo TGA2

And (3) purging gas: nitrogen gas

The heating rate is as follows: 10.0 ℃/min

Temperature range: 25-250 deg.C

4. DVS for dynamic moisture adsorption

The detection adopts SMS DVA Advantage, the humidity is from 0 to 90 percent at 25 ℃, the step is 10 percent, the humidity is from 90 to 95 percent, the step is 5 percent, the judgment standard is that the mass change dM/dT of each gradient is less than 0.002, and the T is_MAXLess than 360min, and circulating for two circles.

5. Ion Chromatography (HPIC): instrument for measuring the position of a moving object

U.S. DionexICS-5000 ion chromatograph; separating the column: IonPac AS14A, detection mode: conducting electricity; leacheate: NaHCO 2₃0.0010M+Na₂CO₃0.0035M; flow rate 1.0mL/min

The structure of the compound is determined by Nuclear Magnetic Resonance (NMR) or/and Mass Spectrometry (MS)And (4) determining. NMR shift (. delta.) at 10^-6The units in (ppm) are given. NMR was measured using a Bruker AVANCE-400 NMR spectrometer using deuterated dimethyl sulfoxide (DMSO-d)₆) Deuterated chloroform (CDCl)₃) Deuterated methanol (CD)₃OD), internal standard Tetramethylsilane (TMS).

Drawings

FIG. 1: an XRPD pattern of form a of the hydrochloride salt.

FIG. 2: an XRPD pattern of form B of the hydrochloride salt.

FIG. 3: TGA profile of form B of the hydrochloride salt.

FIG. 4: XRPD pattern of form C of the hydrochloride salt.

FIG. 5: DSC profile of form C of the hydrochloride salt.

FIG. 6: TGA profile of crystalline form C of the hydrochloride salt.

FIG. 7: XRPD pattern of form a of sulfate.

FIG. 8: TGA profile of form a of sulfate.

FIG. 9: XRPD pattern of form B of sulfate.

FIG. 10: TGA profile of form B of sulfate.

FIG. 11: XRPD pattern of form a of formate.

FIG. 12: DSC profile of form a of formate salt.

FIG. 13: TGA profile of form a of the formate salt.

FIG. 14: DVS profile of form B hydrochloride.

Detailed Description

The present disclosure will be explained in more detail with reference to examples or experimental examples, which are only used to illustrate the technical solutions in the present disclosure, and do not limit the spirit and scope of the present disclosure.

The monitoring of the progress of the reaction in the examples employed thin layer chromatography, the developing agent used for the reaction, the system of eluents for column chromatography used for purifying compounds and the developing agent system for thin layer chromatography including: a: dichloromethane/methanol system, B: in the petroleum ether/ethyl acetate system, the volume ratio of the solvent is adjusted according to different polarities of the compounds, and a small amount of basic or acidic reagents such as triethylamine, acetic acid and the like can be added for adjustment.

Example 1: preparation of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine

The first step is as follows: 3- (6-fluoro-3, 4-dihydronaphthalen-1-yl) azetidine-1-carboxylic acid tert-butyl ester 1c

3-Iodoazetidine-1-carboxylic acid tert-butyl ester 1b (1134.58mg, 4.01mmol, prepared by a known method "Organic Letters, 2014, 16(23), 6160-. Cooled to room temperature, the reaction was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system B to give the title product 1c (700mg), yield: 74.8 percent.

MS m/z(ESI)：304.1[M+1]。

The second step is that: 3- (6-Fluoronaphthalen-1-yl) azetidine-1-carboxylic acid tert-butyl ester 1d

2, 3-dichloro-5, 6-dicyan-p-benzoquinone (336.72mg, 1.48mmol) and 1c (300mg, 0.99mmol) were dissolved in 30mL of a toluene solution, and the mixture was reacted at 80 ℃ for 12 hours. Cooled to room temperature, distilled under reduced pressure, solvent removed by evaporation and the resulting residue purified by thin layer chromatography using developer system B to give the title product 1d (180mg), yield: 60.4 percent.

MS m/z(ESI)：302.2[M+1]。

The third step: 3- (6-fluoronaphthalen-1-yl) azetidine hydrochloride 1e

1d (180mg, 0.60mmol) and 0.5mL of a 4M solution of hydrogen chloride in 1, 4-dioxane were dissolved in 30mL of dichloromethane and the reaction was carried out for 2 hours after the addition was completed. The reaction was concentrated under reduced pressure to give the crude title product 1e (120mg, brown solid) which was directly used for the next reaction without purification.

MS m/z(ESI)：202.1[M+1]。

The fourth step: 3- (6-fluoronaphthalen-1-yl) -N- (6-methoxypyridin-3-yl) azetidine-1-thioamide 1g

Crude 1e (120mg, 0.6) and 1f (99.11mg, 0.60mmol, prepared by the well-known method "Bioorganic & Medicinal Chemistry Letters, 2010, 20(2), 516-. The reaction solution containing 1g of the title product was obtained and used in the next reaction without purification.

MS m/z(ESI)：368.1[M+1]。

The fifth step: (E) -3- (6-fluoronaphthalen-1-yl) -N- (6-methoxypyridin-3-yl) azetidine-1-thioimidate methyl ester 1h

Crude 1g (200mg, 0.54mmol) was dissolved in 50mL tetrahydrofuran solution, cooled to 0 deg.C, and potassium tert-butoxide (183.23mg, 1.63mmol) was dissolved in the solution and reacted for 1 hour after addition. Methyl 4-methylbenzenesulfonate (101.37mg, 0.54mmol) was added to the reaction solution, and the mixture was stirred at room temperature for 12 hours. To the reaction solution was added 50mL of ethyl acetate, washed with water (20mL × 3), the organic phases were combined, the organic phase was distilled under reduced pressure, the solvent was removed by rotation, and the resulting residue was purified by thin layer chromatography with developer system B to give the title product 1h (100mg), yield: 48.2 percent.

MS m/z(ESI)：382.1[M+1]。

And a sixth step: 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine 1

After 1h (100mg, 0.26mmol), trifluoroacetic acid (0.1mL, 0.13mmol) and 2-methoxyacetohydrazide (27.29mg, 0.26mmol) were dissolved in 50mL of tetrahydrofuran, the reaction was carried out for 3 hours under reflux. Cooled to room temperature, the solvent was removed under reduced pressure and the resulting residue was purified by thin layer chromatography using developer system a to give the title product 1(30mg), yield: 26.7 percent.

MS m/z(ESI)：420.1[M+1]。

¹H NMR(400MHz，CD₃OD)δ8.32(s，1H)，7.75-7.84(m，3H)，7.51-7.55(m，2H)，7.43(d，1H)，7.25-7.32(m，1H)，7.00(d，1H)，4.51-4.66(m，1H)，4.35(t，4H)，4.10(t，2H)，4.00(s，3H)，3.26(s，3H)。

Test example 1: determination of inhibitory Activity on human OTR

First, experimental material and instrument

Fluo-4NW calcium assay kit (F36206, invitrogen)

2.MEM(Hyclone，SH30024.01B)

G418 sulfate (Enzo, ALX-380-

4. Fetal bovine serum (GIBCO, 10099)

5. Sodium pyruvate solution (sigma, S8636-100ML)

MEM non-essential amino acid solution (100X) (sigma, M7145-100ML)

Flexstation 3 multifunctional microplate reader (Molecular Devices)

8. Poly-D-lysine 96 well plates, Black/clean (356692, BD)

9. Oxytocin (synthesized by Jier Biochemical Co., Ltd.)

10.pcDNA3.1(invitrogen，V79020)

pcDNA3.1-hOTR (NM-000706) (synthesized and constructed by Jinzhi Biotechnology Ltd. pcDNA3.1 plasmid)

HEK293 cell (cat # GNHu18, cell Bank of Chinese academy of sciences)

Second, the experimental procedure

pcDNA3.1-hOTR plasmid, the use of

3000 transfection reagent is transferred into HEK293 cells; g418 is added for screening every other day, and a monoclonal cell line is selected.

HEK 293/human OTR stable transformants were plated one day in 96-well plates at a density of 25000 cells/well. The next day, the Fluo-4 dye-containing buffer was prepared using the reagents in the Fluo-4NW calcium assay kit, the medium was removed, 100. mu.l of Fluo-4 dye-containing buffer was added to each well, and incubation was carried out at 37 ℃ for 30 minutes. After the time, the plate is moved to room temperature environment for balancing 10And (3) minutes. Compound 10 is prepared⁶、10⁵、10⁴、10³、10²、10¹nM, 1. mu.l per well, and incubation at room temperature for 10 min. The assay was performed using a flexstation 3 microplate reader, and 50. mu.l of 3nM oxytocin polypeptide was automatically added by the machine, reading immediately at 494/516 nM. IC of the Compound₅₀The value can adopt fluorescence values corresponding to different concentrations, and IC is obtained by Graphpad Prism calculation₅₀2nM, indicating that the compound has a significant inhibitory effect on the activity of OTR of human origin.

Test example 2: determination of inhibitory Activity against human V1aR

First, experimental material and instrument

Fluo-4NW calcium assay kit (F36206, invitrogen)

2.MEM(Hyclone，SH30024.01B)

G418 sulfate (Enzo, ALX-380-

4. Fetal bovine serum (GIBCO, 10099)

5. Sodium pyruvate solution (sigma, S8636-100ML)

MEM non-essential amino acid solution (100X) (sigma, M7145-100ML)

Flexstation 3 multifunctional microplate reader (Molecular Devices)

8. Poly-D-lysine 96 well plate, Black/clean (356692, BD)

9. Vasopressin (Tocris, 2935)

10.pcDNA3.1(invitrogen，V79020)

pcDNA3.1-V1aR (NM-000706) (synthesized and constructed by Jinzhi Biotechnology Ltd. pcDNA3.1 plasmid)

HEK293 cell (cat # GNHu18, cell Bank of Chinese academy of sciences)

Second, the experimental procedure

pcDNA3.1-V1aR plasmid was used

One day ahead HEK 293/human V1aR stably transfected cells at 25000Density of individual/well was seeded in 96-well plates. The next day, the Fluo-4 dye-containing buffer was prepared using the reagents in the Fluo-4NW calcium assay kit, the medium was removed, 100. mu.l of Fluo-4 dye-containing buffer was added to each well, and incubation was carried out at 37 ℃ for 30 minutes. After this time, the plate was allowed to equilibrate to room temperature for 10 minutes. Compound 10 is prepared⁶、10⁵、10⁴、10³、10²、10¹nM, 1. mu.l per well, and incubation at room temperature for 10 min. The assay was performed using a flexstation 3 microplate reader, and 50. mu.l of 3nM vasopressin polypeptide was automatically added by the machine, reading immediately at 494/516 nM. IC of the Compound₅₀The value can adopt fluorescence values corresponding to different concentrations, and IC is obtained by Graphpad Prism calculation₅₀4.5nM indicates that the compound inhibits weakly human V1aR activity, indicating a selective inhibition of OTR activity.

Test example 3: determination of inhibitory Activity of Compounds on human V1bR

The inhibitory effect of the compounds of the present disclosure on the activity of the human V1bR protein expressed in HEK 293/human V1bR cells was determined using the following experimental method:

first, experimental material and instrument

Fluo-4NW calcium assay kit (F36206, invitrogen)

2.MEM(Hyclone，SH30024.01B)

G418 sulfate (Enzo, ALX-380-

4. Fetal bovine serum (GIBCO, 10099)

5. Sodium pyruvate solution (sigma, S8636-100ML)

MEM non-essential amino acid solution (100X) (sigma, M7145-100ML)

Flexstation 3 multifunctional microplate reader (Molecular Devices)

8. Poly-D-lysine 96 well plate, Black/clean (356692, BD)

9. Vasopressin (Tocris, 2935)

10.pcDNA3.1(invitrogen，V79020)

pcDNA3.1-V1bR (NM-000706) (synthesized and constructed by Jinzhi Biotechnology Ltd. pcDNA3.1 plasmid)

HEK293 cell (cat # GNHu18, cell Bank of Chinese academy of sciences)

Second, the experimental procedure

pcDNA3.1-V1bR plasmid was used

3000 transfection reagent is transferred into HEK293 cells; g418 was added at the next day to obtain HEK 293/human V1bR pool cell line.

HEK 293/human V1bR pool cells were seeded one day in advance in 96-well plates at a density of 25000 cells/well. The next day, the Fluo-4 dye-containing buffer was prepared using the reagents in the Fluo-4NW calcium assay kit, the medium was removed, 100. mu.l of Fluo-4 dye-containing buffer was added to each well, and incubation was carried out at 37 ℃ for 30 minutes. After this time, the plate was allowed to equilibrate to room temperature for 10 minutes. Compound 10 is prepared⁶、10⁵、10⁴、10³、10²、10¹nM, 1. mu.l per well, and incubation at room temperature for 10 min. The assay was performed using a flexstation 3 microplate reader, and 50. mu.l of 3nM vasopressin polypeptide was automatically added by the machine, reading immediately at 494/516 nM. IC of the Compound₅₀The value can adopt fluorescence values corresponding to different concentrations, and IC is obtained by calculating Graphpad Prism software₅₀At 26 μ M, no significant inhibitory effect of the compound on human V1bR activity was shown, indicating a selective inhibitory effect on OTR activity.

Test example 4: determination of inhibitory Activity of Compounds on human V2R

The inhibitory effect of the compounds of the present disclosure on the activity of human V2R protein expressed in HEK 293/human V2R cells was determined using the following experimental method:

first, experimental material and instrument

cAMP kinetics 2 kit 1, 000 experiments (62AM4PEB, Cisbio)

2.MEM(Hyclone，SH30024.01B)

G418 sulfate (Enzo, ALX-380-

4. Fetal bovine serum (GIBCO, 10099)

5. Sodium pyruvate solution (sigma, S8636-100ML)

MEM non-essential amino acid solution (100X) (sigma, M7145-100ML)

Phearstar multifunctional microplate reader (BMG)

Corning/Costar 384 well non-adsorbing microplate-black NBS plate (4514, Corning)

9. Cell dissociation solution, enzyme-free PBS (13151014-100ml, Thermo Fisher Scientific)

HBSS, calcium, magnesium, phenol Red free (14025-

HEPES, 1M buffer (15630-080, GIBCO)

12.BSA(0219989725，MP Biomedicals)

13.IBMX(I7018-250MG，sigma)

14. Vasopressin (Tocris, 2935)

15.pcDNA3.1(invitrogen，V79020)

pcDNA3.1-V2R (NM-000054) (synthesized and constructed by Jinwei Biotechnology Ltd. pcDNA3.1 plasmid)

HEK293 cell (cat # GNHu18, cell Bank of Chinese academy of sciences)

Second, the experimental procedure

pcDNA3.1-V2R plasmid was used

3000 transfection reagent is transferred into HEK293 cells; g418 was added at the next day to obtain HEK 293/human V2R pool cell line.

1) Cell dissociation:

dissociation of HEK 293/humanized V2R pool cells from the cell culture dish using cell dissociation medium without enzyme, dissociation of the cells into individual cells, termination, pipetting, centrifugation, removal of supernatant, resuspending the cells in assay buffer 1(1 XHBSS +20mM HEPES + 0.1% BSA) and counting, adjusting the cell density to 1250 cells/5. mu.l, i.e., 2.5 x 10⁵/ml。

2) Dispensing of drugs

Compounds were formulated in pure DMSO at a range of concentrations of 20mM, 6.67mM, 2.22mM, 0.74mM, 0.25mM, 0.08mM, 27.4. mu.M, 9.14. mu.M, 3.05. mu.M, 1.02. mu.M, 0.34. mu.M, and 0. mu.M (DMSO). The compound was then made up to 4-fold use concentration using assay buffer 2 (assay buffer 1+1mM IBMX).

Agonist(s): the stock solution of 460. mu.M vasopressin was first made up in DMSO to 2. mu.M and then diluted in assay buffer 2 to 0.5 nM.

And (3) standard substance: the first spot was 20. mu.l stock (2848nM) and the dilutions were made 4-fold sequentially starting from the second spot with assay buffer 1 for a total of 11 concentrations.

3) Adding medicine and incubating:

1. the mixed cells were added to 384-well plates at 5. mu.l/well without changing the tip.

2. Adding 2.5 mul/hole of the prepared compound to be tested and the positive compound, and replacing the gun head.

3.1000rpm for 1min, shaking for 30sec, mixing, and standing and incubating at room temperature for 30 min.

4. Standard curve wells require 5. mu.l/well of assay buffer 2.

5. Adding 2.5 μ l of prepared agonist into each well, replacing the gun head, centrifuging at 1000rpm for 1min, shaking for 30sec, mixing, and standing at room temperature for 30 min.

6. And preparing cAMP-d2 (a component in a cAMP dynamic 2 kit) and Anti-cAMP-Eu-Cryptate (a component in a cAMP dynamic 2 kit) in a dark state according to the ratio of 1: 4, and the cAMP lysate (the component in the cAMP dynamic 2 kit) are mixed evenly. Adding prepared cAMP-d2 liquid 5 μ l/well into each well, adding Anti-cAMP-Eu-Cryptate 5 μ l/well, shaking for 30sec, mixing, and incubating at room temperature in dark for 1 h.

4) Reading a plate: the pherarstar multifunctional microplate reader reads the HTRF signal.

5) Data processing

The data of the experiment were processed using Graphpad Prism to obtain IC₅₀The compound has no obvious inhibition effect on the activity of human V2R, which indicates that the compound has selective inhibition effect on the OTR activity.

Test example 5: determination of the brain-penetrating Activity of Compounds on rats

The brain penetration activity of the compounds of the present disclosure in rats was determined using the following experimental method:

1. experimental materials and instruments

RED Device insertion (Device Inserts) (Thermo Scientific, QL21291110)

API 4000Q-trap linear ion trap mass spectrometer (Applied Biosystems)

LC-30A ultra high pressure liquid chromatography system (Shimadzu)

pH7.4 PBS (100mM, 4 ℃ refrigerator storage)

SD rat, offered by Jersey laboratory animals Co., Ltd, with animal production license number SCXK (Shanghai) 2013-.

2. Operation of laboratory animals

SD rats 4 in each half of male and female, 12/12 light/dark adjustment, constant temperature of 24 + -3 deg.C, humidity of 50-60%, and free access to water. After fasting overnight, the administration was by gavage. The administration dose is 10mg/kg, the administration group is sacrificed (the blood collection amount is 0.5ml) after blood collection for 0.5 h-2 h after administration, the blood sample is placed in a heparinized test tube, blood plasma is separated by centrifugation at 3500rpm for 10min, the blood plasma is marked as blood plasma 1, and the blood plasma is stored at 20 ℃; taking the dead animal, cutting off the head, collecting brain tissue, sucking residual blood with filter paper, recording as brain tissue for 1min, and storing at 0 deg.C after 10 min. Blank plasma and brain tissue 2 were obtained from 3 other animals and treated in the same way as the administration group.

3. Plasma protein binding equilibrium dialysis procedure

3.1 sample preparation

Diluting the drug compound to 50mM with DMSO to obtain stock solution I; transferring a proper amount of stock solution I, and diluting with methanol to obtain 200 mu M diluted stock solution II; 10 μ l of stock solution II is transferred into a 1.5ml Eppendorf tube, 990 μ l of blank plasma is added, and the mixture is uniformly mixed to obtain a2 μ M plasma sample 2 (the DMSO content is less than or equal to 0.2 percent) for measuring the binding rate of the plasma protein at the concentration. The prepared 50. mu.l plasma sample was removed and scored as T₀And storing in a refrigerator at-80 deg.C.

3.2 Experimental procedures

The RED device was inserted into a balanced dialysis tubing set and placed in a 96-well plate. 300. mu.l of the prepared plasma sample 2 containing the analyte and the corresponding blank plasma sample are taken and placed in red-marked wells (plasma chamber). 500 μ l of pH7.4 phosphate buffered saline was placed in another well (buffer chamber) lined with a red label. According to the stepsFor the method, each concentration of each compound is 2-3 samples. After completion, the 96-well bottom plate was covered with a sealing tape (sealing tape), and the whole bottom plate was placed in a thermal mixer and equilibrated at 37 ℃ for 4h at 400 rpm. After incubation, the 96-well bottom plate device was removed from the thermal mixer to complete equilibrium dialysis. Taking 50 μ l of the equilibrated plasma sample or dialysate sample, adding 50 μ l of the corresponding unbalanced drug-free blank phosphate buffer solution or drug-free blank plasma, adding 300 μ l of internal standard (prepared with acetonitrile), vortex mixing for 5min, centrifuging for 10min (4000rpm), and taking the supernatant for LC/MS/MS analysis. T is₀Directly measuring the area ratio of total drug (plasma chamber) and free drug (buffer chamber) to the chromatographic peak of the internal standard substance by the LC/MS/MS method established above without hatching the sample, and calculating the free percentage (f)_{u plasma}％)。

4. Brain tissue protein binding equilibrium dialysis process

Brain tissue protein binding equilibrium dialysis process: blank brain tissue 2 the brain tissue was prepared into blank brain homogenate with ph7.4 PBS according to the dilution factor ═ 11, the compound was added to prepare 2 μ M brain homogenate, the other procedures were the same as those for binding to plasma protein, the ratio of peak areas of total drug (woven homo chamber) and free drug (buffer chamber) to internal standard was determined by established LC/MS method, respectively, and the free percentage (f) was calculated_{u brain hom}％)。

5. Brain permeability test method

1) The drug concentrations in plasma 1 and brain tissue 1, which are total concentrations (C), were determined 0.5h after administration to rats, respectively, using the established LC/MS/MS method_total，pAnd C_total，b)；

2) The free percentage (f) was calculated by measuring the protein binding rate of the compound in rat plasma and brain tissue using an RED Device Inserts apparatus using equilibrium dialysis, respectively_{u plasma}％，f_{u brain}％)；

Percentage free plasma (f)_{u plasma}％)＝C_buffer/C_plasma×100％；

Percent free brain homogenate (f)_{u brain hom}％)＝C_buffer/C_{brain hom}×100％；

Percent brain tissue dissociation (f)_{u brain}％)＝f_{u brain hom}/(Df-(Df-1)*f_{u brain hom}) X is 100%; where Df is 11

3) The blood brain penetration index Kp-unbounded is calculated using the following formula.

6. Test results and discussion

Brain penetration index of a compound

Number of	Kp-unbound	Brain tissue(ng/g)
			1	0.199	435±257

And (4) conclusion: the compounds of the present disclosure have better brain permeability.

Test example 6: pharmacokinetic testing of Compounds

1. Abstract (abstract)

The drug concentrations in the plasma at various times after gavage administration of the compound of example 2, the compound of example 17, the compound of example 34, the compound of example 37, the compound of example 38, the compound of example 39, the compound of example 42 and the compound of example 43 to rats were measured by the LC/MS method using SD male rats as test animals. The pharmacokinetic behavior of the compounds of the present disclosure in rats was studied and their pharmacokinetic profile was evaluated.

2. Test protocol

2.1 test drugs

The compounds of example 2, example 17, example 34, example 37, example 38, example 39, example 42 and example 43.

2.2 test animals

Healthy adult SD rats 24, males divided into 8 groups on average, 3 per group, purchased from shanghai jequirity laboratory animals ltd, animal production license number: SCXK (Shanghai) 2013 and 0006.

2.3 pharmaceutical formulation

A defined amount of drug was weighed out and added 2.5% by volume DMSO and 97.5% by volume 10% solutol HS-15 to make a colorless clear transparent liquid of 0.2 mg/mL.

2.4 administration

SD rats are subjected to gastric lavage after being fasted overnight, the administration dose is 30.0mg/kg, and the administration volume is 10.0 mL/kg.

3. Operation of

The compound of example 2, the compound of example 17, the compound of example 34, the compound of example 37, the compound of example 38, the compound of example 39, the compound of example 42, and the compound of example 43 were administered to rats by gavage, 0.2mL of blood was collected from the orbit before and after administration at 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, 11.0, 24.0 hours, placed in heparinized tubes, centrifuged at 4 ℃ and 3500rpm for 10 minutes to separate plasma, stored at-20 ℃ and fed at 2 hours after administration.

Determining the content of the compound to be tested in rat plasma after the drug with different concentrations is administered by gastric lavage: 50 μ L of rat plasma at each time after administration was taken, 50 μ L of camptothecin as an internal standard solution (100ng/mL), 150 μ L of acetonitrile was added, vortex mixed for 5 minutes, centrifuged for 10 minutes (4000rpm), and 3 μ L of supernatant was taken from plasma samples for LC/MS/MS analysis.

4. Pharmacokinetic parameter results

The pharmacokinetic parameters of the compounds of the disclosure are as follows:

and (4) conclusion: the compounds of the present disclosure are better absorbed and have pharmacokinetic advantages.

Example 2: crystal form A of 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine hydrochloride

Adding 250mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine into 3ml of 95% isopropanol/water, stirring to dissolve the mixture, dropwise adding hydrochloric acid (2mol/L) according to the ratio of 1: 1.1eq (equivalent), continuously stirring, filtering, and drying in vacuum to obtain the product.

Ion Chromatography (HPIC) detection of the resulting product: the chloride ion content was 7.86%, indicating that the molar ratio of compound to hydrochloric acid in the salt was about 1: 1.

The XRPD pattern of this crystalline sample is shown in fig. 1, with a melting peak point around 183.41 ℃ and characteristic peak positions as shown in table 1 below:

TABLE 1

Example 3: crystal form A of hydrochloride

Adding 20mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine into 400ul of ethyl acetate for dissolving, dropwise adding hydrochloric acid (2mol/L) according to a ratio of 1: 1.1eq, continuing stirring, filtering, and drying in vacuum to obtain the product.

Example 4: crystal form B of 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine hydrochloride

Adding 250mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine into 3ml of acetone for dissolving, dropwise adding hydrochloric acid (2mol/L) according to a ratio of 1: 1.1eq, continuing stirring, filtering, and drying in vacuum to obtain the product.

Ion Chromatography (HPIC) detection of the resulting product: the chloride ion content was 7.83%, indicating that the molar ratio of compound to hydrochloric acid in the salt was about 1: 1.

The XRPD pattern of the crystalline sample is shown in fig. 2, the TGA pattern is shown in fig. 3, the melting peak point is around 182.73 ℃, and the characteristic peak positions are shown in table 2 below:

TABLE 2

Example 5: crystal form B of hydrochloride

Adding 20mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine into 400ul of ethanol for dissolving, dropwise adding hydrochloric acid (2mol/L) according to a ratio of 1: 1.1eq, continuing stirring, filtering, and drying in vacuum to obtain the product.

Example 6: crystal form B of hydrochloride

Adding 250mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine into 3ml of acetonitrile for dissolving, dropwise adding hydrochloric acid (2mol/L) according to a ratio of 1: 1.1eq, continuously stirring, filtering,

vacuum drying to obtain the product.

Example 7: c crystal form of 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine hydrochloride

Adding 250mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine into 5ml of methyl tert-butyl ether, dropwise adding hydrochloric acid (2mol/L) according to a ratio of 1: 1.1eq, carrying out temperature rise and drop program stirring within a temperature range of 10-50 ℃, filtering, and carrying out vacuum drying to obtain the product.

Ion Chromatography (HPIC) detection of the resulting product: the chloride ion content was 6.76%, indicating that the molar ratio of compound to hydrochloric acid in the salt was about 1: 1.

The XRPD pattern of this crystalline sample is shown in fig. 4, the DSC pattern is shown in fig. 5, and the TGA pattern is shown in fig. 6, with melting peak points around 170.63 ℃ and characteristic peak positions as shown in table 3 below:

TABLE 3

Example 8: crystal form A of 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine-sulfate

Adding 250mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine into acetonitrile, stirring to dissolve, adding sulfuric acid (2mol/L) according to a ratio of 1: 1.1eq, continuing stirring, filtering, and drying in vacuum to obtain the product.

Ion Chromatography (HPIC) detection of the resulting product: the sulfate content was 32.56%, indicating a molar ratio of compound to sulfuric acid of about 1:2 in the salt.

The XRPD pattern and TGA pattern of the crystalline sample are shown in fig. 8, with characteristic peak positions as shown in table 4 below:

TABLE 4

Example 9: crystal form A of sulfate

Adding 20mg of a compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine into 400ul of ethanol for dissolving, dropwise adding sulfuric acid (2mol/L) according to a ratio of 1: 1.1eq, naturally volatilizing, and filtering to obtain a product.

Example 10: crystal form A of sulfate

Adding 20mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine into 400ul of acetone for dissolving, dropwise adding sulfuric acid (2mol/L) according to a ratio of 1: 1.1eq, continuously stirring, filtering, and drying in vacuum to obtain the product.

Example 11: sulfate crystal form A

Adding 20mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine into 400ul of ethyl acetate for dissolving, dropwise adding sulfuric acid (2mol/L) according to a ratio of 1: 1.1eq, continuously stirring, filtering, and drying in vacuum to obtain the product.

Example 12: crystal form B of 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine-sulfate

Adding 250mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine into 5ml of methyl tert-butyl ether for dissolving, adding sulfuric acid (2mol/L) according to a ratio of 1: 1.1eq, carrying out temperature rise and fall procedures within a temperature range of 10-50 ℃, stirring, filtering, and carrying out vacuum drying to obtain the product.

Ion Chromatography (HPIC) detection of the resulting product: the sulfate content was 18.54%, indicating a molar ratio of compound to sulfuric acid of about 1:1 in the salt.

The XRPD pattern and TGA pattern of the crystalline sample are shown in fig. 10, with characteristic peak positions as shown in table 5 below: -

TABLE 5

Example 13: crystal form A of 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine mesylate

Adding 250mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine into 5ml of methyl tert-butyl ether for dissolving, adding methanesulfonic acid (2mol/L) according to a ratio of 1: 1.1eq, carrying out temperature rise and fall program stirring within a temperature range of 10-50 ℃, filtering, and carrying out vacuum drying at 40 ℃ to obtain the product.

Ion Chromatography (HPIC) detection of the resulting product: the mesylate content was 17.78%, indicating a molar ratio of compound to hydrochloric acid of about 1:1 in the salt.

The XRPD pattern, DSC pattern, TGA pattern of the crystalline sample are shown in the figure, the melting peak point is around 207.46 ℃, and the characteristic peak positions are shown in table 6 below:

TABLE 6

Example 14: hygroscopicity research of hydrochloride A crystal form, hydrochloride B crystal form and mesylate A crystal form

Adopting Surface Measurement Systems adaptation 2, observing the humidity range of 0-95% and the step of 10% at 25 ℃ and humidity from 50%, judging that the mass change dM/dT of each gradient is less than 0.002 and TMAX is less than 360min, and circulating for two circles.

TABLE 7

The sample of the hydrochloride A crystal form is slightly hygroscopic, and the water absorption capacity is increased along with the increase of humidity under the condition of 25 ℃ and between 20.0 percent RH and 80.0 percent RH, the weight change is 0.769 percent, and the moisture absorption weight is increased by less than 2 percent but not less than 0.2 percent. Under normal storage conditions (i.e., 60% humidity at 25 ℃), water absorption is about 1.675%; under accelerated test conditions (i.e., 70% humidity), the water absorption was about 1.738%; under extreme conditions (i.e., 90% humidity), the water absorption is about 1.860%.

During the 0% -95% humidity change, the desorption process and the adsorption process of the sample are basically coincided; x-ray powder diffraction contrast graphs before and after DVS show that crystal forms before and after DVS are not transformed.

Under the condition of 25 ℃, the water absorption capacity of the sample of the hydrochloride B crystal form is increased along with the increase of humidity between 20.0 percent RH and 80.0 percent RH, the weight change is 0.0641 percent, the moisture absorption weight gain is less than 0.2 percent, and the sample has no or almost no moisture absorption; water absorption of about 0.0361% under normal storage conditions (i.e., 60% humidity at 25 ℃); under accelerated test conditions (i.e., 70% humidity), the water absorption was about 0.0446%; under extreme conditions (i.e., 90% humidity), the water absorption is about 0.0947%

During the 0% -95% humidity change, the desorption process and the adsorption process of the sample are basically coincided; the chart of DVS spectrogram, and the contrast chart of X-ray powder diffraction before and after DVS show that the crystal form before and after DVS is not transformed.

Under the condition of 25 ℃, the water absorption amount of a methanesulfonic acid A crystal form sample is increased along with the increase of humidity between 20.0% RH and 80.0% RH, the weight change is 0.1061%, the moisture absorption weight gain is less than 0.2%, and the sample has no or almost no moisture absorption; under normal storage conditions (i.e., 60% humidity at 25 ℃), the water absorption is about 0.0993%; under accelerated test conditions (i.e., 70% humidity), the water absorption was about 0.1154%; under extreme conditions (i.e., 90% humidity), the water uptake is about 0.1755%.

Example 15: study of Crystal form stability

The crystal form of the salt is opened and laid flat, and the stability of the sample under the conditions of illumination (4500Lux), high temperature (40 ℃, 60 ℃) and high humidity (RH 75 percent and RH 92.5 percent) is respectively inspected, wherein the sampling inspection period is 30 days.

TABLE 8

And (4) conclusion: the influence factor experiment shows that: under the conditions of illumination, high temperature of 40 ℃, high humidity of 75 percent and high humidity of 92.5 percent, the A and B of hydrochloride and the A crystal form of mesylate have better physical and chemical stability.

Experimental example 2: long term/accelerated stability

Form B (example 4) was incubated at 25 deg.C, 60% RH and 40 deg.C, 75% RH respectively for stability

TABLE 9

Long term/accelerated stability experiments show that: the hydrochloride crystal form B is placed under the condition of long-term accelerated stability, the crystal form of the hydrochloride crystal form B is not changed, the hydrochloride crystal form B has good physical stability, meanwhile, related substances are not increased, and the hydrochloride crystal form B has excellent chemical stability.

Claims

1. The compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4HPharmaceutically acceptable salts of (E) -1,2, 4-triazol-4-yl) -2-methoxypyridineThe pharmaceutically acceptable salt is selected from hydrochloride, sulfate, methanesulfonate, phosphate, citrate, benzoate or fumarate.

2. The pharmaceutically acceptable salt according to claim 1, wherein the chemical ratio of the compound to the acid molecule is 1:2 to 2: 1.

3. The pharmaceutically acceptable salt of claim 1, wherein the chemical ratio of the compound to the acid molecule is 1:2, 1:1, 2: 1.

4. A process for preparing a pharmaceutically acceptable salt according to claim 1 or 2, comprising: the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4HA step of salifying the (E) -1,2, 4-triazole-4-yl) -2-methoxypyridine with acid.

5. The method according to claim 4, wherein the solvent used for the salt-forming reaction is at least one selected from isopropanol, acetone, methyl tert-butyl ether, acetonitrile, ethanol, ethyl acetate and water.

6. A pharmaceutical composition comprising a pharmaceutically acceptable salt of claim 1 and a pharmaceutically acceptable adjuvant optionally selected from at least one of a pharmaceutically acceptable carrier, diluent or excipient.

7. Use of a pharmaceutically acceptable salt according to claim 1 in the manufacture of a medicament for the treatment or prophylaxis of a disease or condition in which inhibition of oxytocin produces a beneficial effect is known or may be indicated, wherein the disease or condition is selected from the group consisting of sexual dysfunction, pre-partum labor, complications of labor, appetite and eating disorders, benign prostatic hyperplasia, premature labor, dysmenorrhea, congestive heart failure, arterial hypertension, liver cirrhosis, renal hypertension, ocular hypertension, obsessive compulsive disorder and neuropsychiatric disorders.

8. Use according to claim 7, wherein the disease or condition is selected from sexual dysfunction.

9. Use according to claim 7, wherein the disease or condition is selected from hypoactive sexual desire disorder, sexual arousal disorder, orgasm disorder, pain during sexual intercourse disorder and premature ejaculation.

10. Use of a pharmaceutically acceptable salt according to claim 1 for the manufacture of a medicament for antagonising oxytocin.

11. The compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-crystal form A of 1,2, 4-triazole-4-yl) -2-methoxypyridine hydrochloride, characterized in that it has a diffraction angle of 2θThe X-ray powder diffraction pattern expressed by the angle has characteristic peaks at 6.92, 12.54, 15.23, 16.32, 22.70, 27.44 and 28.10.

12. A process for preparing the crystalline form a of claim 11, comprising:

(a) the compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4HAdding the (E) -1,2, 4-triazole-4-yl) -2-methoxypyridine into a solvent (I), stirring for dissolving or heating for dissolving, wherein the solvent (I) is selected from isopropanol/water and ethyl acetate,

(b) dripping hydrochloric acid, stirring and crystallizing.

13. The compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-crystal form B of 1,2, 4-triazole-4-yl) -2-methoxypyridine hydrochloride, characterized in that it has a diffraction angle of 2θThe X-ray powder diffraction pattern has characteristic peaks at 14.62, 15.65, 19.21, 23.66, 24.15, 25.92 and 27.19.

14. The crystalline form B according to claim 13, characterized in that it is at diffraction angle 2θAn X-ray powder diffraction pattern expressed in terms of angles, having the positions 11.76, 14.62, 15.65, 19.21, 23.66, 24.15, 25.92, 27.19, 28.01 and 29.54Characteristic peak.

15. The crystalline form B according to claim 13, characterized in that it is at diffraction angle 2θThe X-ray powder diffraction pattern expressed by the angle has characteristic peaks at 11.76, 14.62, 15.65, 16.43, 18.70, 19.21, 23.66, 24.15, 24.56, 25.60, 25.92, 27.19, 28.01 and 29.54.

16. The crystalline form B according to claim 13, characterized in that it is at diffraction angle 2θThe X-ray powder diffraction pattern at the angle is shown in figure 2.

17. A process for preparing form B of claim 13, comprising:

(a) the compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4HAdding the (E) -1,2, 4-triazole-4-yl) -2-methoxypyridine into a solvent (II), stirring for dissolving or heating for dissolving, wherein the solvent (II) is selected from ethanol, acetone and acetonitrile,

(b) dripping hydrochloric acid, stirring for crystallization, and pulping.

18. The compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine hydrochloride in crystal form C, characterised in that it has a diffraction angle of 2θThe X-ray powder diffraction pattern expressed by the angle has characteristic peaks at 6.94, 21.51, 22.71, 24.99, 25.80, 27.45 and 28.14.

19. A process for preparing the crystalline form C of claim 18, comprising:

(a) the compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4HAdding the (E) -1,2, 4-triazole-4-yl) -2-methoxypyridine into a solvent (III), stirring for dissolving or heating for dissolving, wherein the solvent (III) is selected from methyl tert-butyl ether,

(b) dripping hydrochloric acid, stirring and crystallizing.

20. Chemical combinationThe substance 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-crystal form A of 1,2, 4-triazol-4-yl) -2-methoxypyridine sulfate, characterised in that it has a diffraction angle of 2θThe X-ray powder diffraction pattern expressed by the angle has characteristic peaks at 12.66, 18.50, 19.90, 21.64, 23.61, 24.22 and 26.34.

21. A process for preparing the crystalline form a of claim 20, comprising:

(a) the compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4HAdding the (E) -1,2, 4-triazole-4-yl) -2-methoxypyridine into a solvent (IV), stirring for dissolving or heating for dissolving, wherein the solvent (V) is selected from acetonitrile, ethanol, acetone and ethyl acetate,

(b) dropwise adding sulfuric acid, and stirring for crystallization.

22. The compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-crystal form B of 1,2, 4-triazol-4-yl) -2-methoxypyridine sulfate, characterised in that it has a diffraction angle of 2θThe X-ray powder diffraction pattern expressed by the angle has characteristic peaks at 11.87, 13.22, 14.62, 15.29, 18.49, 22.66 and 23.61.

23. A process for preparing form B of claim 22, comprising:

(a) the compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4HAdding the (E) -1,2, 4-triazole-4-yl) -2-methoxypyridine into a solvent (V), stirring for dissolving or heating for dissolving, wherein the solvent (VI) is selected from methyl tert-butyl ether,

(b) dropwise adding sulfuric acid, and stirring for crystallization.

24. The compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-crystal form A of 1,2, 4-triazol-4-yl) -2-methoxypyridine methanesulfonate characterized by an angle of diffraction of 2θThe X-ray powder diffraction pattern of the angles is 6.66, 16.81, 16.90, 18.92, 19.19, 22.And characteristic peaks at 92 and 24.88.

25. A process for preparing the crystalline form a of claim 24, comprising:

(a) the compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4HAdding the (E) -1,2, 4-triazole-4-yl) -2-methoxypyridine into a solvent (VI), stirring for dissolving or heating for dissolving, wherein the solvent (VII) is selected from ethyl acetate and methyl tert-butyl ether,

(b) methanesulfonic acid is added dropwise, and crystallization is carried out by stirring.

26. A crystalline form according to any of claims 11, 13, 18, 20, 22 or 24, wherein 2 isθThe angular error range is ± 0.30.

27. A crystalline form according to claim 26 characterised in that said 2θThe angular error range is ± 0.20.

28. A pharmaceutical composition comprising the crystalline form of any one of claims 11, 13, 18, 20, 22 or 24 and optionally a pharmaceutically acceptable carrier, diluent or excipient.

29. A pharmaceutical composition prepared from the crystalline form of any one of claims 11, 13, 18, 20, 22, or 24.

30. Use of the crystalline form of any one of claims 11, 13, 18, 20, 22 or 24 for the manufacture of a medicament for the treatment or prevention of a disease or condition known to or which may show a beneficial effect of oxytocin inhibition, characterized in that the disease or condition is selected from the group consisting of sexual dysfunction, pre-partum labor, complications of labor, appetite and eating disorders, benign prostatic hyperplasia, pre-term labor, dysmenorrhea, congestive heart failure, arterial hypertension, liver cirrhosis, renal hypertension, ocular hypertension, obsessive-compulsive disorders and neuropsychiatric disorders.

31. Use according to claim 30, wherein the disease or condition is selected from sexual dysfunction.

32. Use according to claim 30, wherein the disease or condition is selected from hypoactive sexual desire disorder, sexual arousal disorder, orgasm disorder, pain during sexual intercourse disorder and premature ejaculation.

33. Use of the crystalline form of any one of claims 11, 13, 18, 20, 22 or 24 for the manufacture of a medicament for antagonizing oxytocin.