CN111094273B

CN111094273B - Crystal forms of oxytocin receptor inhibitors and methods of making the same

Info

Publication number: CN111094273B
Application number: CN201980004494.1A
Authority: CN
Inventors: 邵启云; 杨俊然; 徐超; 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; Chengdu Suncadia Pharmaceuticals Co Ltd
Priority date: 2018-06-20
Filing date: 2019-06-19
Publication date: 2022-04-12
Anticipated expiration: 2039-06-19
Also published as: WO2019242646A1; TW202016092A; CN111094273A

Abstract

Provides a polymorphic form of an oxytocin receptor (OTR) inhibitor compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) 5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine and a preparation method thereof. Compared with the amorphous form, the polymorphic OTR inhibitor has higher purity and more excellent chemical stability, and has important significance for developing the medicament which is suitable for industrial production and has good biological activity.

Description

Crystal forms of oxytocin receptor inhibitors and methods of making the same

Technical Field

The disclosure provides crystal forms of a compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine 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, and pharmacodynamic experiments show that the compound has a high selective inhibition effect on 0TR, has good brain permeability and can effectively block downstream functions of oxytocin receptors mediated by oxytocin.

Polymorphism refers to the existence of two or more different spatial arrangements of solid substances, thus having different physical and chemical properties. The bioavailability of different crystal forms of the same drug may also differ due to different arrangement modes. Meanwhile, in view of the importance of the crystal form and the stability of the solid medicine to the clinical treatment, the polymorphism research 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 carried out to obtain the crystal form with high purity and stable chemical property, and the method has important significance for developing the medicine which is suitable for industrial production and has good biological activity.

Disclosure of Invention

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, and an X-ray powder diffraction pattern expressed by a diffraction angle 2 theta angle has characteristic peaks at 11.06,14.38,21.81,22.98,24.38,24.80 and 26.88.

In an alternative embodiment, the form a has an X-ray powder diffraction pattern expressed by diffraction angle 2 θ, and the X-ray powder diffraction pattern has characteristic peaks at 11.06,12.10,14.38,18.94,21.09,21.81,22.98,24.38,24.80 and 26.88.

In some embodiments, the form a, having an X-ray powder diffraction pattern expressed in diffraction angle 2 Θ angles, has characteristic peaks at 7.16,10.44,11.06,12.10,14.38,17.00,17.67,18.94,21.09,21.81,22.98,24.38,24.80, 26.88.

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

The disclosure also provides methods 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.

A first process for preparing form a comprising:

(a) adding a 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), and stirring for dissolving or heating for dissolving

(b) Volatilizing;

in the method, the volume (ml) of the solvent (I) is 1-100 times of the weight (g) of the compound, and can be 1,2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 and 100 times; the solvent (I) is selected from at least one of acetone, cyclohexane, ethyl acetate, tetrahydrofuran, methyl tert-butyl ether, 2-butanone, dimethyl sulfoxide, isopropyl acetate, butyl acetate, N-propanol, methyl isobutyl ketone, 1, 4-dioxane, propylene glycol methyl ether, isoamyl alcohol and N, N-dimethylformamide, preferably ethyl acetate, acetone, methyl tert-butyl ether/tetrahydrofuran, acetone/cyclohexane, N-propanol, isopropyl acetate, 2-butanone, dimethyl sulfoxide, methyl isobutyl ketone, 1, 4-dioxane, propylene glycol methyl ether, isoamyl alcohol, water/isopropanol, N-dimethylformamide and butyl acetate.

A second method for preparing form a comprising:

(a) adding a 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 (II),

(b) pulping and filtering;

in the method, the volume (ml) of the solvent (II) 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 (II) is at least one selected from isopropyl ether, n-heptane, n-hexane, methyl tert-butyl ether, methanol, acetonitrile, 1, 4-dioxane, isopropanol, cyclohexane, ethyl acetate and water, preferably isopropanol/water, n-heptane, acetonitrile/cyclohexane, 1, 4-dioxane/cyclohexane, dichloromethane/n-hexane, ethyl acetate/n-hexane, methyl tert-butyl ether, methanol/water and ethyl acetate/n-heptane.

A third method for preparing form a, 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) stirring for crystallization and filtering;

in the method, the volume (ml) of the solvent (II) is 1-20 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 and 20 times; the solvent (III) is at least one selected from acetone, cyclohexane, isopropanol, water, tetrahydrofuran, n-heptane, dichloromethane, ethanol, 1, 4-dioxane and ethyl acetate, preferably isopropyl ether/tetrahydrofuran, isopropanol, acetone/cyclohexane, isopropanol/water, n-heptane/tetrahydrofuran, dichloromethane/n-hexane, ethanol/water, 1, 4-dioxane/cyclohexane and ethyl acetate/n-hexane.

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, and an X-ray powder diffraction pattern expressed by a diffraction angle 2 theta angle has characteristic peaks at 5.54,11.33,16.54,17.10,20.12,21.26 and 22.44.

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 5.54,11.33, 15.67, 16.54,17.10,20.12,21.26,22.44, 25.53.

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.

The disclosure also provides methods of preparing the B crystal form of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine.

A first process for preparing form B 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) volatilizing;

in the method, the volume (ml) of the solvent (IV) is 1-20 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 and 20 times; the solvent (IV) is preferably acetonitrile.

A second method for preparing form B 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) stirring for crystallization and filtering;

in the method, the volume (ml) of the solvent (V) is 1-20 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 and 20 times; the solvent (V) is preferably isopropanol/water in a volume ratio of 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.

A third method for preparing form B, 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) stirring for crystallization and filtering;

in the method, the volume (ml) of the solvent (VI) is 1-20 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 and 20 times; the solvent (VI) is preferably water or tetrahydrofuran/n-hexane, and the volume ratio of the tetrahydrofuran/n-hexane is 0.2-5, and can be 0.2, 0.4, 0.6, 0.8, 1.0, 1.2, 1.4, 1.6, 1.8, 2.0, 2.2, 2.4, 2.6, 2.8, 3.0, 3.2, 3.4, 3.6, 3.8, 4.0, 4.2, 4.4, 4.6, 4.8 and 5.0.

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, and an X-ray powder diffraction pattern expressed by a diffraction angle 2 theta angle has characteristic peaks at 5.63,16.01,17.41,19.02,21.00,23.02,23.84 and 24.28.

In an alternative embodiment, said form C has characteristic peaks at 5.63, 11.55, 16.01,17.41,19.02,21.00,23.02,23.84,24.28, 25.22 in an X-ray powder diffraction pattern expressed in terms of diffraction angle 2 θ.

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 3.

A method of preparing form C comprising:

(b) volatilizing, filtering and vacuum drying.

The disclosure also provides a D 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, and an X-ray powder diffraction pattern expressed by a diffraction angle 2 theta angle has characteristic peaks at 7.78,12.14,12.62,17.25,18.34,19.69 and 25.01.

In an alternative embodiment, said form D, having an X-ray powder diffraction pattern expressed in diffraction angle 2 Θ angles, has characteristic peaks at 7.78, 8.91, 12.14,12.62,17.25,18.34,19.69, 21.26, 25.01, 26.75.

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

Also provided in this disclosure is a method of preparing a crystalline form D of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine.

A method of preparing form D 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 (VII), stirring for dissolving or heating for dissolving,

(b) volatilizing and filtering.

In the method, the volume (ml) of the solvent (VII) is 1-20 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 and 20 times; the solvent (VII) is at least one selected from water, acetone, methanol, ethanol, ethyl acetate, ethanol and acetonitrile, preferably 10% water/acetone, methanol, ethanol, 10% water/methanol, ethyl acetate/ethanol and acetonitrile/methanol.

The present disclosure also provides a pharmaceutical composition prepared from any one of the aforementioned A, B, C or D crystal forms.

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.

Also provided in this disclosure is the use of any one of the aforementioned forms A, B, C or D in the manufacture of a medicament for the treatment or prevention of a disease or condition known or otherwise exhibiting a beneficial effect of oxytocin inhibition, selected from the group consisting of sexual dysfunction, hypoactive sexual desire disorder, sexual arousal disorder, orgasmic disorder, dyspareunia disorder, premature ejaculation, pre-labour, complications of labour, 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, orgasmic disorder, dyspareunia disorder and premature ejaculation.

The present disclosure also provides a use of any one of the aforementioned A, B, C or D crystal forms in the preparation of a medicament for antagonizing oxytocin.

According to the guiding principle of moisture-attracting property of 9103 medicament in 2015 th edition of four parts of Chinese pharmacopoeia and the definition of moisture-attracting 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 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 described in this disclosure has an increased moisture pick-up of 0.1493%, with no or little moisture pick-up, under conditions of 20.0% RH-80% RH.

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

The term "X-ray powder diffraction pattern or XRPD" as used in this disclosure refers to the pattern of X-rays according to bragg formula 2d sin θ ═ n λ (where λ is the wavelength of the X-rays,

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

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.

In the present disclosure, the drying temperature is generally 20 to 100 ℃, preferably 25 to 70 ℃, and the drying may be performed at normal pressure or under reduced pressure (vacuum drying). Preferably, the drying is carried out under reduced pressure.

The chemical and biological agents used in the present disclosure are commercially available.

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

The heating rate is as follows: 10.0 ℃/min

Temperature range: 25-250

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 radiation (λ ═ 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

4. DVS for dynamic moisture adsorption

The detection adopts SMS DVA Advantage, the humidity is from 0-90% at 25 ℃, the step is 10%, the judgment standard is that each gradient mass change dM/dT is less than 0.002, and T is_MAXLess than 360min, and two cyclesAnd (6) looping.

The structure of the compounds is determined by Nuclear Magnetic Resonance (NMR) or/and Mass Spectrometry (MS). NMR shift (. delta.) of 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 compound.

FIG. 2: an XRPD pattern of form B of the compound.

FIG. 3: XRPD pattern of form C of compound.

FIG. 4: an XRPD pattern of form D of the compound.

FIG. 5: DVS profile of form a of compound.

FIG. 6: a comparison of before and after DVS X-ray powder diffraction patterns for form a of the compound.

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

Tert-butyl 3-iodoazetidine-1-carboxylate 1b (1134.58mg, 4.01mmol, prepared by a known method "Organic Letters, 2014, 16(23), 6160-. After cooling, 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 reaction was carried out at 80 ℃ for 12 hours after the addition. 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 mixture, 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 phases were 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 (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 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 oxytocin polypeptide was automatically added by the machine, reading immediately at 494/516 nM. The IC50 value of the compound can be obtained by calculating the IC by Graphpad Prism by adopting fluorescence values corresponding to different concentrations₅₀2nM, 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 plates, 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

HEK 293/human V1aR stably transfected cells were plated at 25000/well density in 96-well plates one day in advance. 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. Detection is carried out by a flexstation 3 microplate readerThe machine automatically added 50. mu.l of 3nM vasopressin polypeptide, 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 plates, 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.

One day ahead HEK 293/human V1bR pool cells at 25000Density per well was 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 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 point is 20: the first stock solution (2848nM) was diluted 4-fold sequentially from the second point 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.

3.1000 rpm 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 the cAMP dynamic 2 kit) in a dark place, and uniformly mixing the cAMP-d2 and the Anti-cAMP-Eu-Cryptate (a component in the cAMP dynamic 2 kit) with cAMP lysate (a component in the cAMP dynamic 2 kit) according to the ratio of 1: 4. 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 IC₅₀4.9 mu M, indicating that the compound has no obvious inhibitory effect on the activity of human V2R, and indicating that the compound has selective inhibitory effect on 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)

pH7.4PBS (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. The treatment method was carried out in the above procedure, with each compound concentration ranging from 2 to 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 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), collectingThe clear solution was analyzed by LC/MS/MS. 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 blank brain homogenate was prepared from brain tissue by using pbs ph7.4 according to the dilution factor of 11, compound was added to prepare 2 μ M brain homogenate, the other procedures were the same as those for binding plasma protein, the ratio of peak area of total drug (woven homo chamber) and free drug (buffer chamber) to internal standard chromatographic peak was measured 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 free (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

Numbering	Kp-unbound	Brain ti ssue(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

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 disclosure in rats was studied and evaluated for their pharmacokinetic profile.

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

100mg of compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine is added into 2ml of methanol and water (V/V, 1: 1), and the mixture is pulped at room temperature, filtered and dried in vacuum to obtain the product. The XRPD pattern of this crystalline sample is shown in fig. 1, with a melting point around 117.03 ℃, an onset melting temperature of 115.18 ℃, and characteristic peak positions as shown in table 1 below:

TABLE 1

Example 3: crystal form A

1.2g of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine was added to 8ml of ethyl acetate and 24ml of n-hexane, heated, stirred and dissolved, stirred, cooled, crystallized, filtered, and vacuum-dried to obtain a product (985mg, yield: 82.1%).

Example 4: crystal form A

100mg of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine was added to 0.5ml of acetone, stirred and dissolved, and after 2ml of cyclohexane was added, the solid was precipitated by natural volatilization, filtered, and dried in vacuum to obtain the product (50mg, yield: 50%).

Example 5: crystal form A

20mg of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine was added to 0.5ml of ethyl acetate, stirred and dissolved, and naturally volatilized to precipitate a solid, which was filtered and dried in vacuum to obtain a product (14mg, yield: 70%).

Example 6: crystal form A

20mg of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine was added to 2ml of methyl t-butyl ether/tetrahydrofuran (V/V, 1: 1) acetone, stirred to dissolve, and naturally volatilized to precipitate a solid, which was filtered and vacuum-dried to obtain a product (15mg, yield: 75%).

Example 7: crystal form A

100mg of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine was added to 1ml of isopropyl ether and 0.5ml of tetrahydrofuran, and the mixture was dissolved by heating with stirring, cooled by stirring to crystallize, filtered, and dried in vacuum to obtain a product (62mg, yield: 62%).

Example 8: crystal form A

100mg of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine was added to 1ml of isopropyl alcohol, heated and stirred to be dissolved, stirred and cooled to be crystallized, filtered, and dried in vacuum to obtain a product (73mg, yield: 73%).

Example 9: crystal form A

50mg of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine was added to 1ml of acetone/cyclohexane (V/V, 1: 5), heated and stirred to be dissolved, stirred and cooled to be crystallized, filtered, and vacuum-dried to obtain a product (28mg, yield: 56%).

Example 10: crystal form A

50mg of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine was added to 1ml of isopropanol/water (V/V, 5: 1), heated and stirred to dissolve, stirred and cooled to crystallize, and then filtered and vacuum-dried to obtain a product (32mg, yield: 64%).

Example 11: crystal form A

50mg of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine was added to 1ml of n-heptane/tetrahydrofuran (V/V, 5: 1), heated and stirred to dissolve, stirred and cooled to crystallize, and then filtered and vacuum-dried to obtain a product (31mg, yield: 62%).

Example 12: crystal form A

50mg of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine was added to 1ml of methylene chloride/n-hexane (V/V, 1: 5), heated and stirred to be dissolved, stirred and cooled to be crystallized, and filtered and vacuum-dried to obtain a product (35mg, yield: 70%).

Example 13: crystal form A

50mg of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine was added to 1ml of ethanol/water (V/V, 5: 1), heated and stirred to be dissolved, stirred and cooled to be crystallized, filtered, and vacuum-dried to obtain a product (27mg, yield: 54%).

Example 14: crystal form A

1g of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine was added to 5ml of 1, 4-dioxane/cyclohexane (V/V, 1: 5), heated and stirred to dissolve, stirred and cooled to crystallize, filtered, and vacuum-dried to obtain a product (876mg, yield: 87.6%).

Example 15: crystal form A

100mg of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine was added to 1ml of ethyl acetate/n-hexane (V/V, 1: 1), slurried, filtered, and dried in vacuo to give a product (63mg, yield: 63%).

Example 16: crystal form A

100mg of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine was added to 1ml isopropyl ether, slurried, filtered, and dried in vacuum to give a product (73mg, yield: 73%).

Example 17: crystal form A

50mg of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine was added to 0.3ml of isopropanol/water (V/V, 3: 1), slurried, filtered, and vacuum-dried to obtain a product (27mg, yield: 54%).

Example 18: crystal form A

50mg of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine was added to 0.3ml of n-heptane, slurried, filtered, and dried in vacuum to give a product (33mg, yield: 66%).

Example 19: crystal form A

50mg of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine was added to 0.3ml of acetonitrile/cyclohexane (V/V, 1: 5), slurried, filtered, and dried in vacuo to give a product (33mg, yield: 66%).

Example 20: crystal form A

50mg of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine was added to 0.3ml of 1, 4-dioxane/cyclohexane (V/V, 1: 5), slurried, filtered, and vacuum-dried to obtain a product (31mg, yield: 62%).

Example 21: crystal form A

50mg of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine was added to 0.3ml of methylene chloride/n-hexane (V/V, 1: 5), followed by beating to give the product (36mg, yield: 72%).

Example 22: crystal form A

Adding 5mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine into 60 mu L of n-propanol, stirring at room temperature for clearing, volatilizing for crystallization, and drying in vacuum to obtain the product.

Example 23: crystal form A

5mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine is added into 60 muL of acetone, stirred and dissolved at room temperature, evaporated and crystallized, and dried in vacuum to obtain the product.

Example 24: crystal form A

Adding 5mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine into 180 mu L of isopropyl acetate, stirring at room temperature for clearing, volatilizing for crystallization, and drying in vacuum to obtain the product.

Example 25: crystal form A

5mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine is added into 500 mu L of methyl tert-butyl ether, undissolved clear at room temperature, heated to 50 ℃ for dissolution, stirred for 30min, cooled to separate out white solid, and dried in vacuum to obtain the product.

Example 26: crystal form A

Adding 5mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine into 40 mu L of 2-butanone, stirring at room temperature for clearing, volatilizing for crystallization, and drying in vacuum to obtain the product.

Example 27: crystal form A

5mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine is added into 60 mu L of dimethyl sulfoxide, stirred and dissolved at room temperature, evaporated, crystallized and dried in vacuum to obtain the product.

Example 28: crystal form A

5mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine is added into 60 muL of methyl isobutyl ketone, stirred and dissolved at room temperature, evaporated, crystallized and dried in vacuum to obtain the product.

Example 29: crystal form A

5mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine is added into 40 mu L of 1, 4-dioxane, stirred and dissolved at room temperature, evaporated and crystallized, and dried in vacuum to obtain the product.

Example 30: crystal form A

Adding 5mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine into 40 mu L of propylene glycol monomethyl ether, stirring at room temperature for clearing, volatilizing for crystallization, and drying in vacuum to obtain the product.

Example 31: crystal form A

Adding 5mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine into 60 mu L of isoamyl alcohol, stirring at room temperature for clearing, volatilizing for crystallization, and drying in vacuum to obtain the product.

Example 32: crystal form A

5mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine is added into 120 mu L of water and isopropanol (V/V, 1: 9), stirred at room temperature for clearing, evaporated and crystallized, and dried in vacuum to obtain the product.

Example 33: crystal form A

5mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine is added into 500. mu.L of ethyl acetate and n-heptane (V/V, 1: 1), stirred at room temperature for clearing, evaporated and crystallized, and dried in vacuum to obtain the product.

Example 34: crystal form A

5mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine is added into 40 mu L N of N-dimethylformamide, stirred at room temperature for clearing, evaporated for crystallization, and dried in vacuum to obtain the product.

Example 35: crystal form A

5mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine is added into 60 mu L of butyl acetate, stirred and dissolved at room temperature, evaporated, crystallized and dried in vacuum to obtain the product.

Example 36: crystal form A

5mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine is added into 500 mu L of cyclohexane, undissolved at room temperature, heated to 50 ℃, stirred for 60min, cooled, pulped and dried in vacuum to obtain the product.

Example 37: b crystal form

100mg of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine was added to 1ml of isopropanol/water (V/V, 10: 1), dissolved by heating, stirred, cooled, crystallized, filtered, and dried in vacuum to give a product (63mg, yield: 63%). The XRPD pattern of this crystalline sample is shown in fig. 2, with characteristic peak positions as shown in table 2 below:

TABLE 2

Example 38: b crystal form

200mg of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine was added to 3ml of acetonitrile, stirred and dissolved, and naturally volatilized to precipitate a solid, which was filtered and dried in vacuum to obtain a product (136mg, yield: 68%).

Example 39: b crystal form

50mg of the compound 5- (3- (3- (6-fluoronaphthalen-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazol-4-yl) -2-methoxypyridine was added to 0.3ml of tetrahydrofuran/cyclohexane (V/V, 1: 2), slurried, filtered, and dried in vacuo to give a product (63mg, yield: 63%).

Example 40: c crystal form

The solid obtained in example 22 was dried in a drying oven for 8 hours to obtain a solid. The XRPD pattern of this crystalline sample is shown in fig. 3, with characteristic peak positions as shown in table 3 below:

TABLE 3

Example 41: d crystal form

100mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine is added into 1ml of acetone and water (V/V, 9: 1) to be dissolved, and the mixture is volatilized at room temperature for crystallization and dried to obtain a solid. The XRPD pattern of this crystalline sample is shown in fig. 4, with a melting point around 86.60 ℃, an onset melting temperature of 79.69 ℃, and characteristic peak positions as shown in table 4 below: -

TABLE 4

Example 42: d crystal form

5mg 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 added into 60 mul of 10% water/methanol for dissolution, and the mixture is volatilized at room temperature for crystallization and dried to obtain a solid.

Example 43: d crystal form

5mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine is added into 50 μ l of methanol to be dissolved, and the solution is volatilized at room temperature for crystallization and dried to obtain a solid.

Example 44: d crystal form

5mg 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 added into 70 mu l of ethanol to be dissolved, and the mixture is volatilized at room temperature for crystallization and dried to obtain a solid.

Example 45: d crystal form

5mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine is added into 60 mul of ethyl acetate/ethanol (V/V, 1: 1) to be dissolved, and the mixture is volatilized at room temperature for crystallization and dried to obtain a solid.

Example 46: d crystal form

5mg of compound 5- (3- (3- (6-fluoronaphthalene-1-yl) azetidin-1-yl) -5- (methoxymethyl) -4H-1,2, 4-triazole-4-yl) -2-methoxypyridine is added into 40 mu l of acetonitrile/methanol (V/V, 1: 1) to be dissolved, and the mixture is volatilized at room temperature for crystallization and dried to obtain a solid.

Example 47: hygroscopicity study of form A

Adopting Surface Measurement Systems for adaptability, observing the humidity range of 0-95% and the step of 10% at 25 ℃ from 50% of humidity, 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 5

And (4) experimental conclusion:

as can be seen from table 5, samples of form a of the compounds of the present disclosure have a water absorption capacity that increases with increasing humidity between 20.0% RH and 80.0% RH at 25 ℃, a weight change of 0.1493% and a weight gain of less than 0.2%, and are free or nearly free of hygroscopicity. Under normal storage conditions (i.e., 60% humidity at 25 ℃), the water absorption is about 0.0809%; under accelerated test conditions (i.e., 70% humidity), the water absorption was about 0.1119%; under extreme conditions (i.e., 90% humidity), the water absorption is about 0.2379%.

During the 0% -95% humidity change, the desorption process and the adsorption process of the sample are basically coincided; the pattern of DVS is shown in figure 5, and the contrast of X-ray powder diffraction before and after DVS shows that the crystal form before and after DVS is not transformed in figure 6(a is XRPD pattern after DVS detection, b is XRPD pattern before DVS detection).

Example 48: study of stability of Crystal form A

Experimental example 1: investigation of influence factors

The crystal form A (example 3) was left open and laid flat, and the stability of the sample was examined under the conditions of light (4500Lux), high temperature (40 ℃ C., 60 ℃ C.), and high humidity (RH 75%, RH 90%), and the sampling period was 20 days.

TABLE 6

The influence factor experiment shows that: the crystal form A has good physical and chemical stability under the conditions of illumination, high temperature of 40 ℃, high temperature of 60 ℃, high humidity of 75 percent and high humidity of 90 percent.

Experimental example 2: long term/accelerated stability

The stability of the form A (example 3) was examined by placing the form A at 25 deg.C, 60% RH and 40 deg.C, 75% RH

TABLE 7

Long term/accelerated stability experiments show that: the crystal form A is placed for 6 months under the condition of long-term accelerated stability, the crystal form is not changed, the physical stability is good, meanwhile, no growth is caused in the aspect of related substances, and the chemical stability is excellent.

Example 49: transformation of crystal form B into crystal form A

After a sample (60mg) of the crystal form B obtained in example 21 is placed at room temperature for two weeks, the sample is subjected to X-ray powder diffraction, and an X-ray powder diffraction spectrum shows that the sample is the crystal form A.

Example 50: transformation of crystal form B into crystal form C

A sample (136mg) of the crystal form B obtained in example 21 is dried for 8 hours in vacuum at room temperature (20-30 ℃) and 0.06Pa, and the sample is subjected to X-ray powder diffraction, and an X-ray powder diffraction spectrum shows that the crystal form C is formed.

Claims

1. The 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 is characterized in that an X-ray powder diffraction pattern expressed by a diffraction angle 2 theta has characteristic peaks at 11.06,14.38,21.81,22.98,24.38,24.80 and 26.88.

2. The crystalline form a according to claim 1, characterized by an X-ray powder diffraction pattern, expressed in terms of diffraction angle 2 Θ angles, having characteristic peaks at 11.06,12.10,14.38,18.94,21.09,21.81,22.98,24.38,24.80, 26.88.

3. The crystalline form a according to claim 1 or 2, characterized by an X-ray powder diffraction pattern, expressed in terms of diffraction angle 2 Θ angles, having characteristic peaks at 7.16,10.44,11.06,12.10,14.38,17.00,17.67,18.94,21.09,21.81,22.98,24.38,24.80, 26.88.

4. Form a according to claim 1, characterized by an X-ray powder diffraction pattern expressed in diffraction angle 2 Θ angles as shown in figure 1.

5. A process for preparing the crystalline form A according to any one of claims 1 to 4, selected from:

the method comprises the following steps:

(a) adding a 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, wherein the solvent (I) is at least one selected from acetone, cyclohexane, ethyl acetate, tetrahydrofuran, methyl tert-butyl ether, 2-butanone, dimethyl sulfoxide, isopropyl acetate, butyl acetate, N-propanol, methyl isobutyl ketone, 1, 4-dioxane, propylene glycol methyl ether, isoamyl alcohol and N, N-dimethylformamide,

(b) volatilizing;

alternatively, method 2:

(a) adding a 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 (II), wherein the solvent (II) is at least one selected from isopropyl ether, n-heptane, n-hexane, acetonitrile, 1, 4-dioxane, isopropanol, cyclohexane, ethyl acetate, methyl tert-butyl ether, methanol or water,

(b) pulping and filtering;

alternatively, method 3:

(a) adding a 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, wherein the solvent (III) is at least one selected from acetone, cyclohexane, isopropanol, water, tetrahydrofuran, n-heptane, dichloromethane, ethanol, 1, 4-dioxane and ethyl acetate,

(b) stirring for crystallization and filtering.

6. The 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 is characterized in that: the X-ray powder diffraction pattern expressed by the angle of diffraction 2 theta has characteristic peaks at 5.54,11.33,16.54,17.10,20.12,21.26 and 22.44.

7. A process for preparing the form B of claim 6 selected from:

the method comprises the following steps:

(a) adding a 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, wherein the solvent (IV) is selected from acetonitrile,

(b) volatilizing;

alternatively, method 2:

(a) adding a 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), and stirring for dissolving or heating for dissolving, wherein the solvent (V) is selected from isopropanol/water;

(b) stirring for crystallization and filtering;

alternatively, method 3:

(a) adding a 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), wherein the solvent (VI) is selected from tetrahydrofuran/n-hexane and water,

(b) pulping and filtering.

8. The 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 is characterized in that: the X-ray powder diffraction pattern expressed by the angle of diffraction 2 theta has characteristic peaks at 5.63,16.01,17.41,19.02,21.00,23.02,23.84 and 24.28.

9. The D 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 is characterized in that: the X-ray powder diffraction pattern expressed by the angle of diffraction 2 theta has characteristic peaks at 7.78,12.14,12.62,17.25,18.34,19.69 and 25.01.

10. A process for preparing the crystalline form D of claim 9, comprising:

(a) adding a 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 (VII), wherein the solvent (VII) is at least one selected from water, acetone, methanol, ethanol, ethyl acetate, ethanol and acetonitrile,

(b) volatilizing and filtering.

11. Form a according to claim 1, characterized in that it has no or almost no hygroscopicity at 20.0% RH-80% RH.

12. A crystalline form according to any of claims 1, 6, 8 or 9, characterized in that the 2 Θ angular error range is ± 0.30.

13. A crystalline form according to any of claims 1, 6, 8 or 9, characterized in that the 2 Θ angular error range is ± 0.20.

14. A pharmaceutical composition comprising the crystalline form of any one of claims 1, 6, 8 or 9 and optionally a pharmaceutically acceptable carrier, diluent or excipient.

15. A pharmaceutical composition prepared from a compound of any one of claims 1, 6, 8, or 9 in crystalline form.

16. Use of a compound of any one of claims 1, 6, 8 or 9 in a crystalline form for the manufacture of a medicament for the treatment or prevention of a disease or condition known to or likely to show a beneficial effect of oxytocin inhibition, selected from the group consisting of sexual dysfunction, hypoactive sexual desire disorder, sexual arousal disorder, orgasmic 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.

17. Use according to claim 16, wherein the disease or condition is selected from sexual dysfunction, sexual arousal disorder, orgasmic disorder, pain disorder during sexual intercourse, and premature ejaculation.

18. Use of a compound of any one of claims 1, 6, 8 or 9 in a crystalline form for the manufacture of a medicament for antagonizing oxytocin.