WO2021143811A1

WO2021143811A1 - Medical salt of tetrazole compound and crystalline form thereof, preparation method therefor and use thereof

Info

Publication number: WO2021143811A1
Application number: PCT/CN2021/072051
Authority: WO
Inventors: 黄建; 祝令建; 李贝; 唐应刚; 尚婷婷; 杜振兴
Original assignee: 上海森辉医药有限公司; 上海盛迪医药有限公司; 江苏恒瑞医药股份有限公司
Priority date: 2020-01-17
Filing date: 2021-01-15
Publication date: 2021-07-22
Also published as: TW202140445A

Abstract

Provided are a medical salt of a tetrazole compound and a crystalline form thereof, a preparation method therefor and use thereof. Specifically, provided are a medical salt (formula I) of 2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazole-1-yl)-1-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyridine-2-yl)propan-2-yl-dihydrogen phosphate and a crystalline form thereof, a preparation method therefor and antifungal use thereof.

Description

Pharmaceutically acceptable salt of tetrazolium compound and its crystal form, preparation method and application

Technical field

The present disclosure belongs to the field of medical technology, and specifically relates to 2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4- (2,2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl dihydrogen phosphate and its crystalline form, preparation method and application.

Background technique

The function of metalloenzymes is highly dependent on the presence of metal ions in the active site of the enzyme. It is recognized that reagents that bind to and inactivate metal ions at the active site greatly reduce the activity of the enzyme. Nature uses this same strategy to reduce the activity of certain metalloenzymes during periods when enzyme activity is not needed. For example, the protein TIMP (tissue inhibitor of metalloproteinases) binds to zinc ions in the active sites of various matrix metalloproteinases, thereby inhibiting enzyme activity. The pharmaceutical industry has used the same strategy in the design of therapeutic agents. For example, the azole antifungal agents fluconazole and voriconazole contain 1-(1,2,4-triazole) group, which exists in the active site of the target enzyme lanosterol demethylase The heme iron binds, thereby inactivating the enzyme. Another example includes zinc-bound hydroxamic acid groups, which have been introduced into most of the published inhibitors of matrix metalloproteinases and histone deacetylases. Another example is the zinc-binding carboxylic acid group, which has been introduced into most of the published angiotensin converting enzyme inhibitors.

VT-1161, the compound 2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2, 2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-ol is an antifungal drug developed by VIAMET, currently in the clinical research stage. Its structure is as follows Shown:

This compound mainly acts on the CYP51 target of fungal cells. Compared with the previous triazole antifungal drugs, it has the advantages of wider antibacterial spectrum, low toxicity, high safety and good selectivity. However, this compound is not suitable for Liquid preparations (including or excluding the parenteral delivery carrier) are used to treat patients in need of such treatment.

2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2,2-trifluoro Ethoxy)phenyl)pyridin-2-yl)propan-2-yl dihydrogen phosphate is a prodrug of VT-1161.

On the other hand, nearly half of the drug molecules are in the form of salts, and salt formation can improve certain undesirable physical, chemical or biological properties of the drug. Relative to 2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2,2- Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl dihydrogen phosphate, it is of great significance to develop a salt with more excellent properties in terms of physical and chemical properties or pharmaceutical properties.

To this end, the present disclosure provides a new pharmaceutically acceptable salt form of a metalloenzyme inhibitor.

Summary of the invention

The present disclosure (The disclosure) provides the compound 2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4 -(2,2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl dihydrogen phosphate pharmaceutically acceptable salt selected from sodium salt and potassium Salt, magnesium salt, lithium salt, ammonium salt, ethanolamine salt and D/L-phenylglycinamide salt, preferably sodium salt.

The pharmaceutically acceptable salts of the present disclosure obtain base addition salts by contacting a neutral form of a compound with a base. In some embodiments, 2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2, 2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate (radical) and cations (including but not limited to sodium ion, magnesium ion, potassium ion, lithium ion, ammonium ion such as ethanolamine The chemical ratio of ions, D/L-phenylglycinamide ions) is 1:1 or 1:2, preferably 1:2.

Some embodiments provide the compound 2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2 The pharmaceutically acceptable salt of ,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl dihydrogen phosphate is:

On the other hand, the compound of the present disclosure 2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2 ,2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl dihydrogen phosphate is a chiral molecule with 1 chiral center and 2 configuration isomers, such as:

(R)-2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2, 2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt (Compound 1).

(R)-2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2, 2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate monosodium salt.

The present disclosure also relates to the preparation of 2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2, The method for 2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl dihydrogen phosphate sodium salt, comprising: compound 2-(2,4-difluorophenyl)-1 ,1-Difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propane The step of reacting -2-yl dihydrogen phosphate with a base selected from sodium hydroxide, sodium bicarbonate, sodium ethoxide and sodium carbonate.

In some embodiments, 2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2, The molar ratio of 2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl dihydrogen phosphate and base is 1:1 or 1:2, preferably 1:2.

In some embodiments, the aforementioned salt formation reaction is carried out in a solvent selected from at least one of ethanol, methanol, tert-butanol, isopropanol, tetrahydrofuran, and acetone.

Further, in an optional embodiment, the method for preparing the aforementioned pharmaceutically acceptable salt further includes the steps of concentration, filtration, drying, solvent volatilization, or stirring and crystallization.

The present disclosure also relates to (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-( 2,2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt (the compound of formula Ia), crystalline form A, expressed by the diffraction angle 2θ angle X- X-ray powder diffraction pattern, there are characteristic peaks at 7.354, 9.280, 9.800, 18.398 and 19.628. In an alternative embodiment, the crystalline form A of the compound of formula I-a has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ, with characteristic peaks at 7.354, 9.280, 9.800, 11.075, 12.942, 18.398 and 19.628.

In an alternative embodiment, the crystalline form A of the compound of formula Ia has an X-ray powder diffraction pattern expressed in diffraction angle 2θ angles at 7.354, 9.280, 9.800, 11.075, 12.942, 14.701, 18.398, 19.628, 21.461 and 22.327 There are characteristic peaks.

In another alternative embodiment, the crystalline form A of the compound of formula I-a has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ as shown in FIG. 1.

On the other hand, the present disclosure also relates to the preparation of (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5 -(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt (compound of formula Ia), the method of crystal form A, including the following step:

(a) Add (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-( 2,2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt was added to solvent (I), stirred and dissolved, and said solvent (I) was selected from methanol Solvent,

(b) Volatilization and crystallization.

On the other hand, the present disclosure also relates to (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5- (4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt (formula Ia compound) crystal form B, with diffraction angle 2θ The indicated X-ray powder diffraction pattern has characteristic peaks at 9.461, 9.948, 12.168, 15.838, 17.146, 18.687 and 20.818.

In an alternative embodiment, the crystalline form B of the compound of formula Ia has an X-ray powder diffraction pattern expressed in diffraction angle 2θ angles, which are characterized at 9.461, 9.948, 12.168, 12.894, 13.674, 15.838, 17.146, 18.687 and 20.818 peak.

In an alternative embodiment, the crystalline form B of the compound of formula Ia has an X-ray powder diffraction pattern expressed in diffraction angle 2θ angles at 9.461, 9.948, 10.707, 12.168, 12.894, 13.674, 15.838, 16.579, 17.146, 18.687, There are characteristic peaks at 20.021 and 20.818.

In another alternative embodiment, the crystalline form B of the compound of formula I-a has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ as shown in FIG. 2.

On the other hand, the present disclosure also relates to the preparation of (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5 -(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt (compound of formula Ia), the method of crystal form B, including the following step:

(a) Add (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-( 2,2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt was added to solvent (II), stirred to dissolve, the solvent (II) was methanol and Mixed solvents of tetrahydrofuran,

(b) Volatilization and crystallization;

or

(a) Add (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-( 2,2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate is dissolved in solvent (III), and the solvent (III) is ethanol,

(b) Add a hydroxide solution, stir and crystallize.

On the other hand, the present disclosure also relates to (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5- (4-(2,2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt (compound of formula Ia), crystalline form C, with a diffraction angle of 2θ The indicated X-ray powder diffraction pattern has characteristic peaks at 8.547, 9.008, 16.043, 22.966, 24.065 and 24.634.

In an alternative embodiment, the crystalline form C of the compound of formula Ia has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ angle, which is characterized at 7.997, 8.547, 9.008, 16.043, 22.465, 22.966, 24.065, 24.634, and 25.944 peak.

In an alternative embodiment, the crystalline form C of the compound of formula Ia has an X-ray powder diffraction pattern expressed as a diffraction angle 2θ angle at 7.997, 8.547, 9.008, 16.043, 22.465, 22.966, 24.065, 24.634, 25.944, 29.826, There are characteristic peaks at 30.761 and 34.937.

In another alternative embodiment, the crystalline form C of the compound of formula I-a has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ as shown in FIG. 3.

On the other hand, the present disclosure also provides the preparation of (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5 -(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt (the compound of formula Ia), the method of crystal form C, including the following step:

(a) Add (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-( 2,2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt is added to the solvent (IV), stirred or heated to dissolve, the solvent (IV) A mixed solution of at least one selected from n-heptane, methyl tert-butyl ether, acetonitrile and isopropanol and water/acetone,

(b) Static devitrification or stirring devitrification;

or,

(R)-2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2 , 2-Trifluoroethoxy) phenyl) pyridin-2-yl) propan-2-yl phosphate disodium salt is added to solvent (V), the solvent (V) is selected from tetrahydrofuran, acetone, methanol, iso At least one of propanol and water,

(b) Volatilization and crystallization;

or,

(a) Add (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-( 2,2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt crystal form A is added to water/isopropanol,

(b) Stir and beat.

On the other hand, the present disclosure also provides (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5- (4-(2,2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt (the compound of formula Ia), crystalline form D, with a diffraction angle of 2θ The indicated X-ray powder diffraction pattern has characteristic peaks at 4.160, 8.388, 9.024, 10.618, 11.568, 20.429 and 22.042.

In an alternative embodiment, the crystalline form D of the compound of formula Ia has an X-ray powder diffraction pattern expressed in diffraction angle 2θ angles, which are characterized at 4.160, 8.388, 9.024, 10.618, 11.568, 14.918, 16.591, 20.429 and 22.042 peak.

In an alternative embodiment, the crystalline form D of the compound of formula Ia has an X-ray powder diffraction pattern expressed in diffraction angle 2θ angles at 4.160, 8.388, 9.024, 10.618, 11.568, 14.918, 16.591, 18.679, 20.429, 20.626, There are characteristic peaks at 22.042 and 22.346.

In another alternative embodiment, the crystalline form D of the compound of formula I-a has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ as shown in FIG. 4.

On the other hand, the present disclosure also relates to the preparation of (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5 -(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt (compound of formula Ia), the method of crystal form D, including the following step:

(a) Add (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-( 2,2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt is added to solvent (VI), stirred and dissolved, and said solvent (VI) is selected from ethanol , Ethanol/water, ethanol/ethyl acetate mixed solution,

(b) Volatilization and crystallization.

On the other hand, the present disclosure also relates to (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5- (4-(2,2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt (the compound of formula Ia), crystalline form E, with a diffraction angle of 2θ The indicated X-ray powder diffraction pattern has characteristic peaks at 5.183, 5.801, 6.904, 7.684, 13.106 and 14.972.

In an alternative embodiment, the crystalline form E of the compound of formula Ia has an X-ray powder diffraction pattern expressed in diffraction angle 2θ angles, which are characterized at 5.183, 5.801, 6.904, 7.684, 13.106, 14.972, 17.177, 20.286, and 22.404 peak.

In another alternative embodiment, the crystalline form E of the compound of formula I-a has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ as shown in FIG. 5.

On the other hand, the present disclosure also relates to the preparation of (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5 -(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt (compound of formula Ia), the method of crystal form E, including the following step:

(a) Add (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-( 2,2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt crystal form A was added to o-xylene,

(b) Stir and beat.

On the other hand, the present disclosure also relates to (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5- (4-(2,2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt (the compound of formula Ia), crystalline form F, with a diffraction angle of 2θ The X-ray powder diffraction pattern shown has characteristic peaks at 6.649 and 9.90.

In another alternative embodiment, the crystalline form F of the compound of formula I-a has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ as shown in FIG. 6.

On the other hand, the present disclosure also relates to the preparation of (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5 -(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt (compound of formula Ia), the method of crystal form F, including the following step:

(a) Add (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-( 2,2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt was added to acetonitrile, stirred to dissolve,

(b) Volatilization and crystallization.

On the other hand, the present disclosure also relates to (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5- (4-(2,2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt (formula Ia compound) crystal form G, with diffraction angle 2θ The indicated X-ray powder diffraction pattern has characteristic peaks at 8.886, 11.313, 12.060, 14.477, 15.433, 15.846 and 19.097.

In an alternative embodiment, the crystalline form G of the compound of formula Ia has an X-ray powder diffraction pattern expressed in diffraction angle 2θ angles, which are characterized at 8.886, 11.313, 12.060, 13.193, 14.477, 15.433, 15.846, 18.010 and 19.097 peak.

In an alternative embodiment, the crystalline form G of the compound of formula Ia has an X-ray powder diffraction pattern expressed in diffraction angle 2θ angles at 8.886, 11.313, 12.060, 13.193, 14.477, 15.433, 15.846, 18.010, 19.097, 20.210, There are characteristic peaks at 20.716 and 22.929.

In another alternative embodiment, the crystalline form G of the compound of formula I-a has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ as shown in FIG. 7.

On the other hand, the present disclosure also relates to the preparation of (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5 -(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt (compound of formula Ia), the method of crystalline form G, including the following step:

(a) Add (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-( 2,2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt is added to the solvent (VII), stirred or heated to dissolve, the solvent (VII) It is a mixed solution of propylene glycol methyl ether and methyl tert-butyl ether or isopropyl ether,

(b) Static devitrification or stirring devitrification;

or,

(R)-2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2 ,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt is added to propylene glycol methyl ether/tetrahydrofuran,

(b) Volatilization and crystallization.

On the other hand, the present disclosure also provides (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5- (4-(2,2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt (the compound of formula Ia), form H, with a diffraction angle of 2θ The indicated X-ray powder diffraction pattern has characteristic peaks at 9.466, 10.638, 12.947, 15.795, 17.053 and 18.653.

In an alternative embodiment, the crystalline form H of the compound of formula I-a has characteristic peaks at 9.466, 9.996, 10.638, 12.947, 15.795, 17.053, 18.653 and 20.113 in the X-ray powder diffraction pattern expressed by diffraction angle 2θ.

In an alternative embodiment, the crystalline form H of the compound of formula Ia has an X-ray powder diffraction pattern expressed in diffraction angle 2θ angles at 9.466, 9.996, 10.638, 12.125, 12.947, 15.795, 17.053, 18.653, 20.113 and 20.759 There are characteristic peaks.

In another alternative embodiment, the crystalline form H of the compound of formula I-a has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ as shown in FIG. 8.

On the other hand, the present disclosure also relates to the preparation of (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5 -(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt (the compound of formula Ia), the method of crystal form H, including the following step:

(a) Add (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-( 2,2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt was added to acetonitrile/methanol (1:1), stirred and dissolved,

(b) Volatilization and crystallization;

or,

(a) Add (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-( 2,2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt crystal form A was added to tetrahydrofuran/ethanol (2:1),

(b) Stir and beat.

On the other hand, the present disclosure also provides (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5- (4-(2,2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt (the compound of formula Ia), crystalline form I, with a diffraction angle of 2θ The indicated X-ray powder diffraction pattern has characteristic peaks at 7.768, 9.525, 10.021, 12.224, 12.982, 17.286 and 21.775.

In an alternative embodiment, the crystalline form I of the compound of formula Ia has an X-ray powder diffraction pattern expressed in diffraction angle 2θ angles, which are characterized at 7.768, 9.525, 10.021, 12.224, 12.982, 15.413, 15.906, 17.286 and 21.775 peak.

In an alternative embodiment, the crystalline form I of the compound of formula Ia has an X-ray powder diffraction pattern expressed in diffraction angle 2θ angles at 7.768, 9.525, 10.021, 12.224, 12.982, 15.413, 15.906, 17.286, 18.849, 20.920, There are characteristic peaks at 21.775 and 22.318.

In another alternative embodiment, the crystalline form I of the compound of formula I-a has an X-ray powder diffraction pattern represented by a diffraction angle of 2θ as shown in FIG. 9.

On the other hand, the present disclosure also relates to the preparation of (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5 -(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt (the compound of formula Ia), the method of crystal form I, including the following step:

(a) Add (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-( 2,2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt was added to 10% water/methanol, stirred to dissolve,

(b) Volatilization and crystallization.

The solvent volume (ml) used in the present disclosure is 1-50 times the compound weight (g), which 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, 50 times.

Further, the preparation method of the crystal form in the present disclosure also includes steps such as filtration, washing or drying. The drying temperature mentioned in the present disclosure is generally 25°C to 100°C, preferably 40°C to 70°C, and it can be dried under normal pressure or under reduced pressure. Preferably, drying is dried under reduced pressure.

The present disclosure also relates to a pharmaceutical composition, which contains the aforementioned pharmaceutically acceptable salt and a pharmaceutical excipient optionally selected from at least one of a pharmaceutically acceptable carrier, diluent or excipient, or is composed of the aforementioned pharmaceutically acceptable salt. It is prepared with a salt and a pharmaceutical excipient optionally selected from at least one of pharmaceutically acceptable carriers, diluents or excipients.

The present disclosure also provides a pharmaceutical composition, which contains crystalline forms A to I of the compound of formula Ia and a pharmaceutical excipient optionally selected from at least one of a pharmaceutically acceptable carrier, diluent or excipient, or It is prepared from the aforementioned crystalline forms A to I of the compound of formula Ia and pharmaceutical excipients optionally selected from at least one of pharmaceutically acceptable carriers, diluents or excipients.

The present disclosure also provides a method for preparing a pharmaceutical composition, which comprises combining the aforementioned pharmaceutically acceptable salt or crystalline form A to I of the compound of formula Ia and optionally selected from pharmaceutically acceptable carriers, diluents or excipients. At least one step of mixing to obtain a composition.

In some embodiments, the unit dose of the pharmaceutical composition is 0.001 mg-1000 mg.

In some embodiments, the pharmaceutical composition contains 0.01-99.99% of the aforementioned pharmaceutically acceptable salt based on the total weight of the composition. In some embodiments, the pharmaceutical composition contains 0.1-99.9% of the aforementioned pharmaceutically acceptable salts. In some embodiments, the pharmaceutical composition contains 0.5% to 99.5% of the aforementioned pharmaceutically acceptable salts. In some embodiments, the pharmaceutical composition contains 1%-99% of the aforementioned pharmaceutically acceptable salts. In some embodiments, the pharmaceutical composition contains 2%-98% of the aforementioned pharmaceutically acceptable salts.

In certain embodiments, the pharmaceutical composition contains 0.01%-99.99% of a pharmaceutically acceptable carrier, diluent or excipient based on the total weight of the composition. In some embodiments, the pharmaceutical composition contains 0.1% to 99.9% of a pharmaceutically acceptable carrier, diluent or excipient. In some embodiments, the pharmaceutical composition contains 0.5% to 99.5% of a pharmaceutically acceptable carrier, diluent or excipient. In some embodiments, the pharmaceutical composition contains 1%-99% of a pharmaceutically acceptable carrier, diluent or excipient. In some embodiments, the pharmaceutical composition contains 2%-98% of a pharmaceutically acceptable carrier, diluent or excipient.

The present disclosure also relates to the use of the aforementioned pharmaceutically acceptable salt or the aforementioned pharmaceutical composition in the preparation of a medicine for regulating the activity of Candida albicans in a subject.

The present disclosure also relates to the use of the aforementioned crystalline forms A to I of the compound of formula I-a in the preparation of a medicament for regulating the activity of Candida albicans in a subject.

On the other hand, the present disclosure also relates to the use of the aforementioned pharmaceutically acceptable salt or the aforementioned pharmaceutical composition in the preparation of a medicament for the treatment of Candida albicans-related disorders or diseases. In some embodiments, wherein the disease or condition is systemic fungal infection or onychomycosis.

The present disclosure also relates to the use of the aforementioned crystalline forms A to I of the compound of formula I-a in the preparation of a medicament for the treatment of Candida albicans related disorders or diseases. In some embodiments, wherein the disease or condition is systemic fungal infection or onychomycosis.

The present disclosure also relates to the use of the aforementioned pharmaceutically acceptable salt or the aforementioned pharmaceutical composition in the preparation of a medicine for inhibiting the activity of metalloenzymes. In some embodiments, wherein the metalloenzyme is lanosterol demethylase (CYP51).

The present disclosure also relates to the use of the aforementioned crystal forms A to I of the compound of formula I-a in the preparation of a medicine for inhibiting the activity of metalloenzymes. In some embodiments, wherein the metalloenzyme is lanosterol demethylase (CYP51).

The present disclosure also relates to the use of the aforementioned pharmaceutically acceptable salt or the aforementioned pharmaceutical composition in the preparation of a medicament for the treatment of a metalloenzyme-mediated disorder or disease, wherein the metalloenzyme-mediated disorder or disease is demethylated by lanosterol Enzyme (CYP51) mediated. In some embodiments, wherein the disease or condition is an infectious disease. In other embodiments, wherein the disease or condition is superficial fungal infection, mucosal fungal infection, systemic fungal infection, or onychomycosis.

The present disclosure also relates to the use of the aforementioned crystal forms A to I of the compound of formula Ia in the preparation of a medicament for the treatment of metalloenzyme-mediated disorders or diseases, wherein the metalloenzyme-mediated disorders or diseases are depleted by lanosterol. Methylase (CYP51) mediated. In some embodiments, wherein the disease or condition is an infectious disease. In other embodiments, wherein the disease or condition is superficial fungal infection, mucosal fungal infection, systemic fungal infection, or onychomycosis.

The present disclosure also relates to the aforementioned pharmaceutically acceptable salt or the aforementioned pharmaceutical composition, which is used to modulate the activity of Candida albicans in a subject.

The present disclosure also relates to the aforementioned crystalline forms A to I of the compound of formula I-a, which are used to modulate the activity of Candida albicans in a subject.

The present disclosure also relates to the aforementioned pharmaceutically acceptable salt or the aforementioned pharmaceutical composition, which is used for the treatment of Candida albicans-related disorders or diseases. In some embodiments, wherein the disease or condition is systemic fungal infection or onychomycosis.

The present disclosure also relates to the aforementioned crystalline forms A to I of the compound of formula I-a, which are used for the treatment of Candida albicans-related disorders or diseases. In some embodiments, wherein the disease or condition is systemic fungal infection or onychomycosis.

The present disclosure also relates to the aforementioned pharmaceutically acceptable salt or the aforementioned pharmaceutical composition, which is used to inhibit metalloenzyme activity. In some embodiments, wherein the metalloenzyme is lanosterol demethylase (CYP51).

The present disclosure also relates to the aforementioned crystalline forms A to I of the compound of formula I-a, which are used to inhibit metalloenzyme activity. In some embodiments, wherein the metalloenzyme is lanosterol demethylase (CYP51).

The present disclosure also relates to the aforementioned pharmaceutically acceptable salt or the aforementioned pharmaceutical composition for the treatment of metalloenzyme-mediated conditions or diseases, wherein the metalloenzyme-mediated conditions or diseases are mediated by lanosterol demethylase (CYP51) guide. In some embodiments, wherein the disease or condition is an infectious disease. In other embodiments, wherein the disease or condition is superficial fungal infection, mucosal fungal infection, systemic fungal infection, or onychomycosis.

The present disclosure also relates to the aforementioned crystalline forms A to I of the compound of formula Ia, which are used for the treatment of metalloenzyme-mediated conditions or diseases, wherein the metalloenzyme-mediated conditions or diseases are mediated by lanosterol demethylase (CYP51) guide. In some embodiments, wherein the disease or condition is an infectious disease. In other embodiments, wherein the disease or condition is superficial fungal infection, mucosal fungal infection, systemic fungal infection, or onychomycosis.

The present disclosure also relates to a method for regulating the activity of Candida albicans in a subject, which comprises administering to the subject a therapeutically effective amount of the aforementioned pharmaceutically acceptable salt or the aforementioned pharmaceutical composition or the aforementioned crystalline form A to I of the compound of formula Ia .

The present disclosure also relates to a method for treating Candida albicans-related disorders or diseases, which comprises administering to a subject in need thereof a therapeutically effective amount of the aforementioned pharmaceutically acceptable salt or the aforementioned pharmaceutical composition or the aforementioned pharmaceutical composition or the aforementioned formula Ia Compound crystalline forms A to I. In some embodiments, wherein the disease or condition is systemic fungal infection or onychomycosis.

The present disclosure also relates to a method for treating Candida albicans-related disorders or diseases, which comprises administering to a subject in need thereof a therapeutically effective amount of the aforementioned pharmaceutically acceptable salt or the aforementioned pharmaceutical composition or the aforementioned crystalline form A of the compound of formula Ia To I. In some embodiments, wherein the disease or condition is systemic fungal infection or onychomycosis.

The present disclosure also relates to a method for inhibiting metalloenzyme activity, which comprises administering a therapeutically effective amount of the aforementioned pharmaceutically acceptable salt or the aforementioned pharmaceutical composition or the aforementioned crystalline form A to I of the compound of formula I-a to a subject in need thereof. In some embodiments, wherein the metalloenzyme is lanosterol demethylase (CYP51).

The present disclosure also relates to a treatment of a metalloenzyme-mediated disorder or disease, which comprises administering to a subject in need thereof a therapeutically effective amount of the aforementioned pharmaceutically acceptable salt or the aforementioned pharmaceutical composition or the aforementioned crystalline form A to of the compound of formula Ia I, wherein the metalloenzyme-mediated condition or disease is mediated by lanosterol demethylase (CYP51). In some embodiments, wherein the disease or condition is an infectious disease. In other embodiments, wherein the disease or condition is superficial fungal infection, mucosal fungal infection, systemic fungal infection, or onychomycosis.

On the other hand, the hydrogen in the functional group of the aforementioned pharmaceutically acceptable salt of the present disclosure is deuterated to obtain the corresponding deuterated compound. The deuterated compound retains selectivity and potential comparable to hydrogen analogs; the deuterium bond is more stable, making "ADME" That is, the "toxic pharmacokinetics" are different, thereby providing clinically beneficial effects. Toxic pharmacokinetics refers to the absorption, distribution, metabolism and excretion of foreign chemicals by the body. Examples of the deuterated compounds of the present disclosure are as follows:

XRPD is an X-ray powder diffraction test: the measurement is carried out using a BRUKER D8 X-ray diffractometer. The specific information collected: Cu anode (40kV, 40mA), Cu-Kα1 rays

Kα2 rays

Kβ rays

Scanning range (2θ range): 3-50, scanning step length 0.02, slit width (collimator) 1.0mm.

The "2θ or 2θ angle" mentioned in the present disclosure refers to the diffraction angle, and θ is the Bragg angle, in degrees or degrees; the error range of each characteristic peak 2θ is ±0.20, which can be -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, 0.20.

The "interplanar spacing or interplanar spacing (d value)" mentioned in the present disclosure means that the spatial lattice selects three non-parallel unit vectors a, b, and c connecting two adjacent lattice points. The lattice is divided into juxtaposed parallelepiped units, called interplanar spacing. The space lattice is divided according to the determined parallelepiped unit lines to obtain a set of linear grids, which are called spatial lattices or lattices. Lattice and crystal lattice use geometric points and lines to reflect the periodicity of the crystal structure. For different crystal planes, the interplanar spacing (that is, the distance between two adjacent parallel crystal planes) is different; the unit is

Or angstrom.

The "differential scanning calorimetry or DSC" mentioned in the present disclosure refers to the measurement of the temperature difference and heat flow difference between the sample and the reference material during the temperature rise or constant temperature process of the sample to characterize all physical changes related to the thermal effect and Chemical changes to obtain the phase change information of the sample.

The "pharmaceutical composition" of the present disclosure means a mixture containing one or more of the compounds described herein or their physiologically/pharmaceutically acceptable salts or prodrugs and other chemical components, as well as other components such as physiologically/pharmaceutically acceptable Carriers and excipients. The purpose of the pharmaceutical composition is to promote the administration to the organism, which is beneficial to the absorption of the active ingredient and thus the biological activity.

The crystalline forms described in the present disclosure include, but are not limited to, the solvate of the compound of formula I-a, and the complex formed by combining the compound of formula I-a with a solvent, and the solvent includes but is not limited to water.

The numerical values in this disclosure are measured by instruments, and there is a certain degree of error. Generally speaking, plus or minus 10% are within a reasonable error range. Of course, it is necessary to consider the context in which the value is used. For example, the chemical ratio of the compound to the acid or base molecule in the present disclosure is that the error after the measurement does not change by more than plus or minus 10%, and it can be plus or minus 9%, plus or minus. 8%, plus or minus 7%, plus or minus 6%, plus or minus 5%, plus or minus 4%, plus or minus 3%, plus or minus 2%, or plus or minus 1%, preferably plus or minus 5%. The drying temperature mentioned in the present disclosure is generally 25°C to 100°C, preferably 40°C to 70°C, and it can be dried under normal pressure or under reduced pressure. It is preferable to dry under reduced pressure.

In the chemical structure of the compound described in the present disclosure, the bond

Indicates that the configuration is not specified, that is, if there are chiral isomers in the chemical structure, the bond

Can be

or

Or both

and

Two configurations.

The reagents used in this disclosure are commercially available.

The test conditions of the instrument used in the experiment in this disclosure:

X-ray Powder Diffraction (XRPD)

(1) Instrument model: Bruker D8 Discover A25 X-ray powder diffractometer

Ray: monochromatic Cu-Kα rays (λ=1.5418)

Scanning method: θ/2θ, scanning range: 3-50 ^°

Voltage: 40KV, current: 40mA

The structure of the compound can be determined by nuclear magnetic resonance (NMR) or/and mass spectrometry (MS). The NMR shift (δ) is given in units of 10-6 (ppm). NMR was measured with Bruker AVANCE-400 nuclear magnetic instrument, and the solvent was deuterated dimethyl sulfoxide (DMSO-d ₆ ), deuterated chloroform (CDCl ₃ ), deuterated methanol (CD ₃ OD), and the internal standard was four Methylsilane (TMS).

For MS measurement, use FINNIGAN LCQAd (ESI) mass spectrometer (manufacturer: Thermo, model: Finnigan LCQ advantage MAX).

High performance liquid chromatography (HPLC) analysis uses Agilent HPLC 1200DAD, Agilent HPLC 1200VWD and Waters HPLC e2695-2489 high pressure liquid chromatograph.

Description of the drawings

Figure 1: XRPD spectrum of the crystalline form A of the compound of formula I-a.

Figure 2: XRPD spectrum of the crystalline form B of the compound of formula I-a.

Figure 3: XRPD spectrum of the crystalline form C of the compound of formula I-a.

Figure 4: XRPD spectrum of the crystalline form D of the compound of formula I-a.

Figure 5: XRPD spectrum of the crystalline form E of the compound of formula I-a.

Figure 6: XRPD spectrum of the crystalline form F of the compound of formula I-a.

Figure 7: XRPD spectrum of the crystalline form G of the compound of formula I-a.

Figure 8: XRPD spectrum of the crystalline form H of the compound of formula I-a.

Figure 9: XRPD spectrum of the crystalline form I of the compound of formula I-a.

Figure 10: Amorphous XRPD spectrum of the compound of formula I-a.

Detailed ways

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

Example 1:

(R)-2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2, 2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt (compound 1)

(R)-2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2 ,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate (compound 1a, prepared according to the method of patent WO2013110002, 0.28g, 0.46mmol, 1.0eq) and ethanol (5mL ) Add to the reaction flask and stir evenly. A solution of NaOH (36.90 mg, 2.0 eq) dissolved in water (1 mL) was added dropwise to the above reaction flask, stirring was continued for 2 h, and concentrated to obtain compound 1, 300 mg of white solid.

The X-ray powder diffraction test showed that there is no sharp diffraction peak in the XRPD spectrum, as shown in FIG. 10.

Ms:608.10[M-2Na+3H] ⁺ .

Ion chromatography detected that the sodium ion content was 6.23%.

Example 2: (R)-((2-(2,4-Difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4 -(2,2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)prop-2-yl)oxy)methyl phosphate disodium salt (compound 2)

Under ice cooling, NaH (58mg, 0.87mmol) was added to the reaction flask, 1.5mL of N,N-dimethylformamide and 0.6mL of tetrahydrofuran were added, followed by iodine (38mg, 0.15mmol), and then Compound 2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2,2-tri Fluoroethoxy)phenyl)pyridin-2-yl)propan-2-ol (2b, prepared according to the method of patent WO2013110002, 158mg, 0.3mmol) tetrahydrofuran (1ml) solution was added to the reaction solution, stirred for 1-4h Then add compound 2a (519mg, 2.01mmol) in tetrahydrofuran (1ml) solvent to the reaction, stir until the reaction is complete, 10% ammonium chloride aqueous solution quenches the reaction, extract, concentrate and drain, and use the crude product 2c directly. One-step reaction, Ms: 750.0[M+H] ⁺ .

Under ice-bath cooling, add trifluoroacetic acid (0.5mL) to the crude product 2c (300mg) in dichloromethane (2mL), stir until the reaction is complete, and after concentration, the target compound 2d, 82mg, Ms was separated by high performance liquid phase separation. :638.0[M+H] ⁺ .

Add compound 2d (0.29g, 0.46mmol, 1.0eq) and ethanol (5mL) obtained in the previous step into the reaction flask, stir, and add NaOH (36.90mg, 2.0eq) water (1ml) solution dropwise to the aforementioned reaction solution , Stirred for 2-5 h, and concentrated to obtain 2,313 mg of the target compound.

Ms:638.10[M-2Na+3H] ⁺ .

Example 3:

According to the method of Example 1, (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)- was prepared using potassium hydroxide as a base 1-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate dipotassium salt (Compound 3)

Ion chromatography detected that the potassium ion content was 9.57%.

Detected by X-ray powder diffraction, its characteristic peak positions are shown in Table 1

Table 1

Example 4:

According to the method of Example 1, (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)- was prepared with calcium hydroxide as a base. 1-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate calcium salt (Compound 4)

Detected by X-ray powder diffraction, its characteristic peak positions are shown in Table 2

Table 2

Example 5:

Prepare (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4 -(2,2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate magnesium salt (compound 5)

Example 6:

Prepare (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4 -(2,2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate amine salt (compound 6)

Detected by X-ray powder diffraction, its characteristic peak positions are shown in Table 3

table 3

Test Example 1: Water solubility and solution stability

1. Preparation of reagents

(1) 0.2mol/L NaH ₂ PO ₄ ·2H ₂ O solution; weigh 31.2g of NaH ₂ PO ₄ ·2H ₂ O (or 27.6g of NaH ₂ PO ₄ ·H ₂ O) and add water to 1000ml to dissolve.

(2) 0.2mol/L Na ₂ HPO ₄ solution: Weigh 71.632g of Na ₂ HPO ₄ ·12H ₂ O (or 53.6g of Na ₂ HPO ₄ ·7H ₂ O or Na ₂ HPO ₄ ·2H ₂ O 35.6 g) Add water to 1000ml to dissolve.

pH=4.0: Adjust the pH of _{0.2mol/L NaH 2} PO ₄ to 4.0 with 8.5% phosphoric acid solution

pH=7.0: Measure 39ml of 0.2mol/L NaH ₂ PO ₄ and 61ml of 0.2mol/L Na ₂ HPO _{4 and} mix it, then finely adjust the pH;

pH=7.4: Mix 19ml of 0.2mol/L NaH ₂ PO ₄ and 81ml of 0.2mol/L Na ₂ HPO ₄ , and then finely adjust the pH;

pH=9.0: Adjust the pH of _{0.2mol/L Na 2} HPO _{4 to 9.0 with 1M sodium hydroxide solution}

2. Test method

Accurately weigh appropriate amount of compound 1 and VT-1161 to be tested, add small amounts to the solution several times and stir to wait for the compound to dissolve, and determine the content of the compound in the solution.

in conclusion:

1) Solvency:

Compound 1 has a solubility greater than 4mg/ml under the condition of pH=4; under the condition of pH=7, the solubility is greater than 5mg/ml; under the condition of pH=9, the solubility is greater than 6mg/ml.

Compound 2 has a solubility of 1.0 mg/ml at pH=4; at pH=7, its solubility is 0.97 mg/ml; at pH=9, its solubility is 0.99 mg/ml.

Compound 3 has a solubility greater than 4mg/ml under the condition of pH=4; under the condition of pH=7.4, the solubility is greater than 5mg/ml; under the condition of pH=9, the solubility is greater than 6mg/ml.

Compound 4 has a solubility of 0.81mg/ml under the condition of pH=4; under the condition of pH=7.4, the solubility is 0.87mg/ml; under the condition of pH=9, the solubility is 0.95mg/ml.

Compound 5 has a solubility of 1.67 mg/ml under the condition of pH=4; under the condition of pH=7.4, the solubility is 1.89mg/ml; under the condition of pH=9, the solubility is 1.78mg/ml.

Compound 6 has a solubility of 0.89mg/ml under the condition of pH=4; under the condition of pH=7.4, the solubility is 0.77mg/ml; under the condition of pH=9, the solubility is 3.0mg/ml.

Compared with other salts, the solution solubility of potassium salt or sodium salt is superior, and the solubility of compound 1 is better than that of compound 2, which provides solubility guarantee for the preparation of injections.

2) In terms of solution stability, dissolve compounds 1, 3, 4, 5 and 6 in different pH solutions respectively, and detect the content of related substances in the solution at the beginning, 1h, 2h, 4h and 7h. The relevant data are shown in Table 4 to 8.

Table 4: Solution stability of compound 1 in different pH solutions

Table 5: Solution stability of compound 3 in different pH solutions

Table 6: Solution stability of compound 4 in different pH solutions

Table 7: Solution stability of compound 5 in different pH solutions

Table 8: Solution stability of compound 6 in different pH solutions

Note: N.A is not detected; N.D is not detected.

Test Example 2: Solid stability

Weigh about 1 mg of the sample into a vial, place it in a vacuum bag and vacuum, then put it into a container with color-changing silica gel, seal it, and prepare two copies in parallel. Prepare enough copies according to the sampling time point, and place them at 2-8°C and room temperature. The solid stability data of compound 1 and compound 3 are shown in Tables 9 and 10, respectively. It is not difficult to see that the disodium salt (compound 1) exhibits excellent samples at room temperature and at 2-8°C, and the increase in the main degradation impurities is small. It is expected that the drug can be ensured in the later preparation process. Long-term shelf life under harsh storage conditions.

Table 9: Compound 1 solid stability data

Table 10: Compound 3 solid stability data

Test Example 3: In vivo pharmacokinetic experiment in dogs

Preparation of administration formulation: Compound 1, accurately weigh an appropriate amount of the test substance, add an appropriate volume of physiological saline, stir or sonicate uniformly to obtain a clear dosing solution with a concentration of 0.3 mg/mL for intravenous administration.

experimental design:

Time point of blood collection:

0.083h, 0.25h, 0.5h, 1h, 2h, 4h, 8h, 24h, 48h before and after administration, a total of 10 blood sampling points.

Sample collection and disposal:

Blood is collected through the jugular vein or other suitable methods, and each sample is collected about 1 mL, K ₂ EDTA is anticoagulated, and placed on ice after collection, and the plasma is separated by centrifugation within 1 hour (centrifugal force 2200 g, centrifugation 10 min, 2-8°C). The collected plasma samples were stored in a refrigerator at -80°C before analysis.

Biological analysis and data processing:

Detect the concentration of compound 1 and VT-1161 in plasma samples, evaluate the accuracy of quality control samples while analyzing the samples, and require more than 66% of the quality control samples to have an accuracy of 80-120%.

Through the blood drug concentration data at different time points, WinNonlin is used to calculate the pharmacokinetic parameters, such as AUC(0-t), T _1/2 , Cmax, Tmax and MRT.

Table 11

Conclusion: The compound of formula I-a can be effectively transformed into the active ingredient of the female drug in dogs.

Example 7:

Take 15 mg of the compound of formula I-a prepared by referring to the method in Example 1, add 0.15ml 10% water/acetone solution, stir to dissolve, add 0.75ml isopropyl ether, stir to crystallize, filter, and dry to obtain a solid. It is amorphous by X-ray powder diffraction.

Example 8:

Take 15 mg of the compound of formula I-a prepared by referring to the method in Example 1, add 0.15 ml of methanol solution, stir to dissolve, add 0.75 ml of methyl tert-butyl ether, stir to crystallize, filter, and dry to obtain a solid. It is amorphous by X-ray powder diffraction.

Example 9:

Take 15 mg of the compound of formula I-a prepared by referring to the method in Example 1, add 0.15ml 10% ethyl acetate solution, stir to dissolve, add 0.75ml n-heptane, stir to crystallize, filter, and dry to obtain a solid. It is amorphous by X-ray powder diffraction.

Example 10:

Take 5 mg of the compound of formula I-a prepared by referring to the method in Example 1, add 0.02 ml of methanol, stir to dissolve, and slowly volatilize at room temperature to obtain a solid. After X-ray powder diffraction detection, the XRPD spectrum is shown in Figure 1, and its characteristic peak positions are shown in Table 12, which is defined as crystal form A.

Table 12

Example 11:

Take 15 mg of the compound of formula I-a prepared by referring to the method in Example 1, add 0.15 ml of methanol, stir to dissolve, add 0.75 ml of tetrahydrofuran, and slowly volatilize at room temperature to obtain a solid. After X-ray powder diffraction detection, the XRPD spectrum is shown in Figure 2, and its characteristic peak positions are shown in Table 13, which is defined as crystal form B.

Table 13

Example 12:

Add 15 mg of compound 1a (prepared according to the method of patent WO2013110002) into 0.44 ml of ethanol, stir to dissolve, add 56 ul of 1N sodium hydroxide solution, precipitate a solid after stirring, filter and dry to obtain a solid. The crystal form B was detected by X-ray powder diffraction.

Example 13:

Take 5mg of crystal form A prepared by referring to the method in Example 10, add 0.5ml 10% water/isopropanol solution, beat at room temperature for 3 days, filter, and dry to obtain a solid. After X-ray powder diffraction detection, the XRPD spectrum is shown in Figure 3, and the characteristic peak positions are shown in Table 14, which is defined as crystal form C.

Table 14

Example 14:

Take 5mg of the compound of formula I-a prepared by referring to the method in Example 1, add 0.02ml 10% water/acetone, stir to dissolve, and slowly volatilize at room temperature to obtain a solid. It was found to be crystal form C by X-ray powder diffraction.

Example 15:

Take 5 mg of the compound of formula I-a prepared by referring to the method in Example 1, add 0.24 ml of methanol/water (1:1), stir to dissolve, and slowly volatilize at room temperature to obtain a solid. It was found to be crystal form C by X-ray powder diffraction.

Example 16:

Take 15 mg of the compound of formula I-a prepared by referring to the method in Example 1, add 0.15 ml of water, stir to dissolve, add 0.75 ml of isopropanol, and slowly volatilize at room temperature to obtain a solid. It was found to be crystal form C by X-ray powder diffraction.

Example 17:

Take 15 mg of the compound of formula I-a prepared by referring to the method in Example 1, add 0.15ml 10% water/acetone, stir to dissolve, add 0.75ml tetrahydrofuran, and slowly volatilize at room temperature to obtain a solid. It was found to be crystal form C by X-ray powder diffraction.

Example 18:

Take 15 mg of the compound of formula I-a prepared by referring to the method in Example 1 and add it to 0.15 ml of 10% water/acetone, stir to dissolve, add 0.75 ml of isopropanol, stir to crystallize, filter, and dry to obtain a solid. It was found to be crystal form C by X-ray powder diffraction.

Example 19:

Take 15 mg of the compound of formula I-a prepared by referring to the method in Example 1 and add 15 mg of 10% water/acetone to 0.15 ml of 10% water/acetone, stir to dissolve, add 0.75 ml of acetonitrile, stir to crystallize, filter, and dry to obtain a solid. It was found to be crystal form C by X-ray powder diffraction.

Example 20:

Take 15 mg of the compound of formula I-a prepared according to the method in Example 1 and add it to 0.15 ml of 10% water/acetone, stir to dissolve, add 0.75 ml of methyl tert-butyl ether, stir to crystallize, filter, and dry to obtain a solid. It was found to be crystal form C by X-ray powder diffraction.

Example 21:

Take 15 mg of the compound of formula I-a prepared by referring to the method in Example 1 and add it to 0.15 ml of 10% water/acetone, stir to dissolve, add 0.75 ml of n-heptane, stir to crystallize, filter, and dry to obtain a solid. It was found to be crystal form C by X-ray powder diffraction.

Example 22:

Take 5 mg of the compound of formula I-a prepared by referring to the method in Example 1, add 0.34 ml of ethanol, stir to dissolve, and slowly volatilize at room temperature to obtain a solid. After X-ray powder diffraction detection, the XRPD spectrum is shown in Figure 4, and the characteristic peak positions are shown in Table 15, which is defined as crystal form D.

Table 15

Example 23:

Take 5 mg of the compound of formula I-a prepared by referring to the method in Example 1, add 0.26 ml of ethyl acetate/ethanol (1:1), stir to dissolve, and slowly volatilize at room temperature to obtain a solid. The crystal form D was detected by X-ray powder diffraction.

Example 24:

Take 5mg of crystal form A prepared by referring to the method in Example 10 and add it to 0.5ml of 7% water/ethanol, stir insoluble at room temperature, raise and lower the temperature at 45-5°C, stir and crystallize at 5°C, filter, and dry to obtain a solid. The crystal form D was detected by X-ray powder diffraction.

Example 25:

Take 5mg of crystal form A prepared by referring to the method of Example 10, add it to 0.5ml o-xylene, beat at room temperature, filter, and dry to obtain a solid. After X-ray powder diffraction detection, the XRPD spectrum is shown in Figure 5, and the characteristic peak positions are shown in Table 16, which is defined as crystal form E.

Table 16

Example 26:

Take 5 mg of crystal form A prepared by referring to the method in Example 10 and add it to 0.5 ml of acetonitrile, stir and filter the insoluble matter, and the filtrate is slowly volatilized at room temperature to obtain a solid. After X-ray powder diffraction detection, the XRPD spectrum is shown in Figure 6, and its characteristic peak positions are shown in Table 17, which is defined as crystal form F.

Table 17

Example 27:

Take 5 mg of the compound of formula I-a prepared by referring to the method in Example 1 and add it to 0.02 ml of propylene glycol methyl ether, stir to dissolve, and slowly volatilize at room temperature to obtain a solid. After X-ray powder diffraction detection, the XRPD spectrum is shown in Fig. 7, and the characteristic peak positions are shown in Table 18, which is defined as crystal form G.

Table 18

Example 28:

Take 15 mg of the compound of formula I-a prepared according to the method in Example 1 and add it to 0.15 ml of propylene glycol methyl ether, stir to dissolve, add 0.75 ml of tetrahydrofuran, and slowly volatilize at room temperature to obtain a solid. It was found to be crystal form G by X-ray powder diffraction.

Example 29:

Take 15 mg of the compound of formula I-a prepared according to the method in Example 1 and add it to 0.15 ml of propylene glycol methyl ether, stir to dissolve, add 0.75 ml of methyl tert-butyl ether, stir to crystallize, filter, and dry to obtain a solid. It was found to be crystal form G by X-ray powder diffraction.

Example 30:

Take 15 mg of the compound of formula I-a prepared according to the method in Example 1 and add it to 0.15 ml of propylene glycol methyl ether, stir to dissolve, add 0.75 ml of isopropyl ether, stir to crystallize, filter, and dry to obtain a solid. It was found to be crystal form G by X-ray powder diffraction.

Example 31:

Take 15 mg of the compound of formula I-a prepared by referring to the method in Example 1 and add it to 0.02 ml of acetonitrile/methanol (1:1), stir to dissolve, and slowly volatilize at room temperature to obtain a solid. After X-ray powder diffraction detection, the XRPD spectrum is shown in Figure 8, and the characteristic peak positions are shown in Table 19, which is defined as crystal form H.

Table 19

Example 32:

Take 5 mg of crystal form A prepared by referring to the method in Example 10, add 0.5 ml of tetrahydrofuran/ethanol (2:1), stir at room temperature to make a slurry, filter, and dry to obtain a solid. It was found to be crystal form H by X-ray powder diffraction.

Example 33:

Take 5 mg of the compound of formula I-a prepared by referring to the method in Example 1 and add it to 0.08 ml of 10% water/methanol, stir to dissolve, and slowly volatilize at room temperature to obtain a solid. After X-ray powder diffraction detection, the XRPD spectrum is shown in Figure 9, and its characteristic peak positions are shown in Table 20, which is defined as crystal form I.

Table 20

Test Example 4: Research on Moisture Diversion

Using Surface Measurement Systems advantage 2, at 25°C, the humidity starts from 50%, the humidity range is 0%-95%, and the step is 10%. The judgment standard is that the mass change dM/dT of each gradient is less than 0.002%, The humidity gradient running time TMAX is 360min, and the cycle is two times.

Table 21

Test Example 5: Study on the stability of crystal form

1) Lay the crystal form of the sodium salt flat, and examine the stability of the sample under the conditions of light (4500 Lux), high temperature (40°C, 60°C), and high humidity (RH 75%, RH 92.5%), and take a sample The inspection period is 30 days.

Table 22

Conclusion: The influencing factor experiments show that: under the conditions of light, high temperature 40℃ and 60℃, high humidity 75% and 92.5% for 30 days, the crystal form C of formula Ia compound has good physical stability; under high humidity and light conditions , Good chemical stability.

2) Long-term/accelerated stability: The crystal form A and crystal form C of the compound of formula I-a are placed at -20°C, 4°C, 25°C/60%RH and 40°C/75%RH respectively to investigate the stability.

Table 23

Conclusion: Long-term accelerated experiments show that: under the conditions of -20℃, 4℃, 25℃/60RH and 40℃/75RH for 3 months, the compound of formula Ia crystal form C has good physical stability, and the compound of formula Ia crystal form A Poor physical stability. The results of the long-term accelerated experiment for 3 months show that the crystal forms A and C of the compound of formula I-a have good chemical stability at -20°C and 4°C.

Test Example 6: Take the compound of formula I-a crystal form C and compound 3 (prepared by the method in Example 2), respectively, and place them under the conditions of light, high temperature 40°C and 60°C, high humidity 75% and 92.5% to investigate the stability.

Table 24

Conclusion: The experiment of influencing factors shows that under the conditions of light, high temperature 40℃ and 60℃, high humidity 75% and 92.5% for 30 days, the physical and chemical stability of sodium salt is good, and the physical and chemical stability of potassium salt is slightly Difference.

Claims

Compound 2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2,2,2-tri A pharmaceutically acceptable salt of fluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl dihydrogen phosphate, characterized in that the pharmaceutically acceptable salt is a sodium salt.
The pharmaceutically acceptable salt according to claim 1, wherein the 2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl) The chemical ratio of -1-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate to cation is 1:1 or 1 :2, preferably 1:2.
The pharmaceutically acceptable salt according to claim 1 or 2, characterized in that it is (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetra Azol-1-yl)-1-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)prop-2-yl phosphate disodium salt
A method for preparing the pharmaceutically acceptable salt according to any one of claims 1-3, comprising the compound 2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazole) -1-yl)-1-(5-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl dihydrogen phosphate and alkali The base is selected from sodium hydroxide, sodium bicarbonate, sodium carbonate and sodium ethoxide.
The method according to claim 4, wherein the reaction is carried out in a solvent, and the solvent is selected from at least one of ethanol, methanol, tert-butanol, isopropanol, tetrahydrofuran, and acetone.
A pharmaceutical composition comprising the pharmaceutically acceptable salt according to any one of claims 1 to 3 and a pharmaceutical excipient optionally selected from at least one of a pharmaceutically acceptable carrier, diluent or excipient, or It is prepared from the pharmaceutically acceptable salt according to any one of claims 1 to 3 and a pharmaceutical excipient optionally selected from at least one of pharmaceutically acceptable carriers, diluents or excipients.
The use of the pharmaceutically acceptable salt according to any one of claims 1 to 3 or the pharmaceutical composition according to claim 6 in the preparation of a medicament for regulating the activity of Candida albicans in a subject.
The use of the pharmaceutically acceptable salt according to any one of claims 1 to 3 or the pharmaceutical composition according to claim 6 in the preparation of a medicament for the treatment of Candida albicans related disorders or diseases, said diseases or disorders Preferably, it is systemic fungal infection or onychomycosis.
The crystalline form A of the pharmaceutically acceptable salt according to claim 3, characterized in that the X-ray powder diffraction pattern expressed by diffraction angle 2θ has characteristic peaks at 7.354, 9.280, 9.800, 18.398 and 19.628, preferably There are characteristic peaks at 7.354, 9.280, 9.800, 11.075, 12.942, 18.398, and 19.628, more preferably at 7.354, 9.280, 9.800, 11.075, 12.942, 14.701, 18.398, 19.628, 21.461, and 22.327, with characteristic peaks at best diffraction. The X-ray powder diffraction pattern represented by the angle 2θ angle is shown in Fig. 1, where the error range of each characteristic peak 2θ is ±0.2.
The crystalline form C of the pharmaceutically acceptable salt according to claim 3, wherein the X-ray powder diffraction pattern represented by the diffraction angle 2θ has characteristic peaks at 8.547, 9.008, 16.043, 22.966, 24.065 and 24.634 , Preferably have characteristic peaks at 7.997, 8.547, 9.008, 16.043, 22.465, 22.966, 24.065, 24.634 and 25.944, more preferably at 7.997, 8.547, 9.008, 16.043, 22.465, 22.966, 24.065, 24.634, 25.944, 29.826, 30.761 and There are characteristic peaks at 34.937, and the best X-ray powder diffraction pattern represented by the diffraction angle 2θ is shown in Figure 3, where the error range of each characteristic peak 2θ is ±0.2.
A method for preparing the crystal form C according to claim 10, selected from:

method one:

a) Add (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2 , 2,2-Trifluoroethoxy) phenyl) pyridin-2-yl) propan-2-yl phosphate disodium salt is added to the solvent (IV), stirred to dissolve or heated to dissolve, the solvent (IV) is selected From at least one of n-heptane, methyl tert-butyl ether, acetonitrile and isopropanol mixed with water/acetone,

b) Static devitrification or stirring devitrification;

Or, method two:

a) Add (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-(5-(4-(2 ,2,2-Trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt is added to the solvent (V), the solvent (V) is selected from tetrahydrofuran, acetone, methanol , At least one of isopropanol and water,

b) Volatilization and crystallization;

Or, method three:

a) The (R)-2-(2,4-difluorophenyl)-1,1-difluoro-3-(1H-tetrazol-1-yl)-1-( 5-(4-(2,2,2-trifluoroethoxy)phenyl)pyridin-2-yl)propan-2-yl phosphate disodium salt crystalline form A is added to water/isopropanol,

b) Mixing and beating.
A pharmaceutical composition comprising the crystal form A according to claim 9 or the crystal form C according to claim 10 and at least one of pharmaceutically acceptable carriers, diluents or excipients Pharmaceutical excipients, or from at least one of the crystalline form A according to claim 9 or the crystalline form C according to claim 10 and optionally from at least one of a pharmaceutically acceptable carrier, diluent or excipient Preparation of pharmaceutical excipients.
A method for preparing a pharmaceutical composition, comprising the crystal form A of claim 9 or the crystal form C of claim 10 and at least one of a pharmaceutically acceptable carrier, diluent or excipient optionally This is the step of mixing to obtain the composition.
Use of the crystal form A according to claim 9 or the crystal form C according to claim 10 or the pharmaceutical composition according to claim 12 in the preparation of a medicament for regulating the activity of Candida albicans in a subject.
The use of the crystal form A according to claim 9 or the crystal form C according to claim 10 or the pharmaceutical composition according to claim 12 in the preparation of a medicament for the treatment of Candida albicans related disorders or diseases, so The disease or condition is preferably systemic fungal infection or onychomycosis.