CN112679441A

CN112679441A - Crystal form of apraxitant, preparation method and application thereof

Info

Publication number: CN112679441A
Application number: CN202011152311.1A
Authority: CN
Inventors: 贾强; 马天华; 杨金金; 王云中; 杨兵兵; 黄法; 赖广建; 华莹
Original assignee: Seasons Biotechnology Taizhou Co ltd
Current assignee: Seasons Biotechnology Taizhou Co ltd
Priority date: 2019-10-18
Filing date: 2020-10-18
Publication date: 2021-04-20

Abstract

The invention relates to an aplitant polymorphic form, a preparation method and application thereof. Specifically, the invention relates to a polymorphism of an N- [5- (4-bromophenyl) -6- [2- [ (5-bromo-2-pyrimidinyl) oxy ] ethoxy ] -4-pyrimidinyl ] sulfonamide compound, a preparation method and application thereof. The invention discloses six different crystal forms of apraxitant, a preparation method, a pharmaceutical composition of apraxitant and application of the apraxitant in preparation of medicines for treating diseases such as hypertension and pulmonary hypertension.

Description

Crystal form of apraxitant, preparation method and application thereof

Technical Field

The invention relates to the field of drug crystal technology and pharmacy. In particular to an aplitant polymorphic form, a preparation method and application thereof. Further relates to a polymorphic form of N- [5- (4-bromophenyl) -6- [2- [ (5-bromo-2-pyrimidinyl) oxy ] ethoxy ] -4-pyrimidinyl ] sulfonamide, and a preparation method and application thereof.

Background

Polymorphism of drugs is common during drug development. Different crystal forms of the same medicament may have obvious difference in aspects of appearance, solubility, melting point, dissolution rate, bioavailability and the like, so that the stability, bioavailability and curative effect of the medicament are influenced, and the phenomenon is particularly obvious in the aspect of oral solid preparations. The polymorphism of the drug is one of important factors influencing the quality and clinical efficacy of the drug, and it is very critical and important to discover the polymorphism of the drug (including anhydrates, hydrates and solvates) as much as possible and to preferentially select the crystal form with better bioavailability, storage stability, easy processing and purification for drug development.

Apraxitant (the english name aprocitent) is an oral dual endothelin receptor antagonist, which has achieved very positive results in phase II clinical trials for the treatment of hypertension. The chemical name of the apraxitant is as follows: n- [5- (4-bromophenyl) -6- [2- [ (5-bromo-2-pyrimidinyl) oxy]Ethoxy radical]-4-pyrimidinyl]A sulfonamide. The chemical formula is: c₁₆H₁₄Br₂N₆O₄S; molecular weight: 546.19, respectively; the chemical structural formula is as follows:

the current reports on crystalline forms of apraxitant are limited to the discovery of apraxitant in the examples of patents WO2009024906 and EP2907811, which mention that crystalline forms can be obtained, but the patents do not disclose any data or spectra relating to the crystalline forms. Meanwhile, when the crystallization experiment in the above patent is repeated, the crystallization process is found to have the problems that a pure single crystal form cannot be stably obtained and acid degradation impurities are generated by the apraxitant during the acidification crystallization of sulfuric acid.

Based on the defects of the prior art and the important significance of the drug crystal form, the existing crystal form of the apraxitant, especially the existing polymorphism, needs to be comprehensively researched so as to accurately obtain the crystal form condition of the apraxitant, and a proper crystal form is preferentially selected for drug development so as to ensure the drug quality and obtain better clinical treatment effect.

Disclosure of Invention

Aiming at the defects of the prior art, the invention mainly aims to provide crystal forms of the apraxitant with different crystal forms, a preparation method and application thereof.

According to an object of the present invention, the present invention provides crystalline form I of apraxitant (hereinafter referred to as "crystalline form I").

The X-ray powder diffraction of the crystal form I has characteristic peaks at the following 2 theta angle positions by using Cu-K alpha radiation: 9.7 +/-0.2 degrees, 17.8 +/-0.2 degrees, 19.8 +/-0.2 degrees and 23.3 +/-0.2 degrees.

The form I also has characteristic peaks at the following 2 theta angular positions: 11.6 +/-0.2 degrees, 15.2 +/-0.2 degrees, 15.5 +/-0.2 degrees, 16.8 +/-0.2 degrees, 18.5 +/-0.2 degrees, 21.3 +/-0.2 degrees, 24.7 +/-0.2 degrees, 25.4 +/-0.2 degrees, 27.1 +/-0.2 degrees and 28.9 +/-0.2 degrees.

The form I also has characteristic peaks at the following 2 theta angular positions: 14.3 +/-0.2 degrees, 21.8 +/-0.2 degrees, 22.7 +/-0.2 degrees, 23.0 +/-0.2 degrees, 28.3 +/-0.2 degrees, 29.2 +/-0.2 degrees, 30.4 +/-0.2 degrees, 31.6 +/-0.2 degrees, 33.8 +/-0.2 degrees and 39.7 +/-0.2 degrees.

According to the purpose of the invention, the invention also provides a preparation method of the crystalline form I of the apraxitant, which adopts any one of the following four methods to prepare the crystalline form I of the apraxitant:

the method comprises the following steps of 1) adding a good solvent into the apraxitant to enable the apraxitant to be dissolved clearly at high temperature, cooling and crystallizing, and then separating and drying precipitated crystals to obtain the crystal form I;

the good solvent is selected from one or more of C3-C6 ester, C3-C4 ketone, chloroform, tetrahydrofuran and methyl tetrahydrofuran;

the high temperature is selected from 40 ℃ to the boiling point of a solvent system, and is preferably 50-80 ℃; the temperature after cooling is selected from 0-30 ℃, and room temperature is preferred;

in the solution, the amount of the apraxitant is 0.1-1 times, preferably 0.5-1 times, and more preferably 0.8-1 times of the solubility of the apraxitant in the solvent system at high temperature;

method 2) adding a good solvent into the apraxitant to dissolve the apraxitant at high temperature, adding a certain amount of poor solvent at high temperature to keep the system still dissolved, cooling and crystallizing, and then separating and drying the precipitated crystal to obtain the crystal form I;

the good solvent is selected from one or more of C3-C6 ester, C3-C4 ketone and acetonitrile;

the poor solvent is selected from one or more of n-hexane, n-heptane and toluene;

the high temperature is selected from 40 ℃ to the boiling point of a solvent system, and is preferably 50-80 ℃; the temperature after cooling is 0-30 ℃, and room temperature is preferred;

the volume ratio of the poor solvent to the good solvent is 0.1-100: 1, preferably 0.5-50: 1, and more preferably 0.5-20: 1;

method 3) adding a pre-mixed good solvent and poor solvent mixed solution into the apraxitant, dissolving the system at a high temperature to be clear, cooling and crystallizing, and then separating and drying the precipitated crystal to obtain the crystal form I;

method 4) adding a good solvent into the apraxitant to dissolve the apraxitant, adding water at room temperature, forcibly crystallizing while stirring, and then separating and drying the precipitated crystal to obtain the crystal form I;

the good solvent is one or a mixture of N, N-dimethylformamide, N-dimethylacetamide and dimethyl sulfoxide;

in the solution, the amount of the apraxitant is 0.1-1 times, preferably 0.5-1 times, and more preferably 0.8-1 times of the solubility of the apraxitant in the solvent system at room temperature;

the volume ratio of the water to the good solvent is 0.1-100: 1, preferably 0.5-50: 1, and more preferably 0.5-20: 1;

the present invention also provides crystalline form II of apraxitant (hereinafter referred to as "crystalline form II"), which is an object of the present invention.

The X-ray powder diffraction of said form II has characteristic peaks at the following 2 Θ angular positions using Cu-ka radiation: 9.2 +/-0.2 degrees, 16.0 +/-0.2 degrees, 18.6 +/-0.2 degrees and 22.3 +/-0.2 degrees.

The form II also has characteristic peaks at the following 2 Θ angular positions: 11.0 +/-0.2 degrees, 20.2 +/-0.2 degrees, 24.1 +/-0.2 degrees, 24.7 +/-0.2 degrees, 26.0 +/-0.2 degrees and 29.6 +/-0.2 degrees.

The form II also has characteristic peaks at the following 2 Θ angular positions: 17.8 +/-0.2 degrees, 19.6 +/-0.2 degrees, 22.8 +/-0.2 degrees, 25.4 +/-0.2 degrees, 27.1 +/-0.2 degrees, 28.1 +/-0.2 degrees, 29.2 +/-0.2 degrees, 32.3 +/-0.2 degrees, 34.0 +/-0.2 degrees, 36.6 +/-0.2 degrees and 37.7 +/-0.2 degrees.

According to an object of the present invention, the present invention also provides a process for the preparation of crystalline form II of apraxitant:

dissolving apraxitant in dichloromethane at high temperature, adding a certain amount of poor solvent at high temperature to keep the system still dissolved, cooling and crystallizing, and separating and drying the precipitated crystal to obtain the crystal form II;

the poor solvent is selected from C5-C8 alkane, preferably n-hexane, cyclohexane and n-heptane, more preferably n-heptane;

the elevated temperature is selected from the boiling point of the solvent system; the temperature after cooling is selected from 0-30 ℃, and room temperature is preferred;

the volume ratio of the dichloromethane to the poor solvent is 0.1-100: 1, preferably 0.5-50: 1, and more preferably 0.5-20: 1;

the present invention also provides, according to the object of the present invention, crystalline form III of apraxitant (hereinafter referred to as "crystalline form III").

The X-ray powder diffraction of form III has characteristic peaks at the following 2-theta angular positions using Cu-ka radiation: 7.6 +/-0.2 degrees, 19.4 +/-0.2 degrees, 21.8 +/-0.2 degrees and 24.3 +/-0.2 degrees.

The form III also has characteristic peaks at the following 2 Θ angular positions: 9.5 +/-0.2 degrees, 15.6 +/-0.2 degrees, 17.1 +/-0.2 degrees, 23.4 +/-0.2 degrees, 26.6 +/-0.2 degrees and 28.4 +/-0.2 degrees.

The form III also has characteristic peaks at the following 2 Θ angular positions: 18.5 +/-0.2 degrees, 20.0 +/-0.2 degrees, 20.5 +/-0.2 degrees, 22.3 +/-0.2 degrees, 22.8 +/-0.2 degrees, 23.1 +/-0.2 degrees, 25.2 +/-0.2 degrees, 25.6 +/-0.2 degrees, 29.1 +/-0.2 degrees, 31.4 +/-0.2 degrees, 32.7 +/-0.2 degrees and 39.7 +/-0.2 degrees.

According to the purpose of the invention, the invention also provides a preparation method of the crystalline form III of the apraxitant, which adopts any one of the following four methods to prepare the crystalline form III of the apraxitant:

the method comprises the following steps of 1) adding a good solvent into the apraxitant to enable the apraxitant to be dissolved clearly at high temperature, adding a poor solvent to enable a system to still keep the apraxitant at high temperature, cooling and crystallizing, and separating and drying precipitated crystals to obtain the crystal form III;

the good solvent is selected from ether solvents, preferably one or more of 1, 4-dioxane, tetrahydrofuran, methyltetrahydrofuran and glycol dimethyl ether;

the poor solvent is selected from C5-C8 alkane and aromatic hydrocarbon solvent, the C5-C8 alkane is preferably n-hexane, cyclohexane and n-heptane, the aromatic hydrocarbon solvent is preferably toluene, xylene, trimethylbenzene and halogenated benzene, and the poor solvent can be selected from single solvent or a plurality of mixed solvents;

the high temperature is selected from 40 ℃ to the boiling point of a solvent system, and is preferably 60-90 ℃; the temperature after cooling is selected from 0-30 ℃, and room temperature is preferred;

in the solution, the amount of the apraxitant is 0.1-1 time, preferably 0.5-1 time, and more preferably 0.8-1 time of the solubility of the apraxitant in the good solvent at room temperature;

method 2) adding a pre-mixed good solvent and poor solvent mixed solution into the apraxitant, dissolving the system at a high temperature to be clear, cooling and crystallizing, and separating and drying the precipitated crystal to obtain the crystal form III;

method 3) adding a good solvent into the apraxitant to dissolve the apraxitant at high temperature, adding a poor solvent at high temperature to perform forced crystallization, cooling and continuing stirring for a period of time, separating and drying the precipitated crystals to obtain the crystal form III;

the poor solvent is selected from C5-C8 alkane, preferably n-hexane, cyclohexane and n-heptane;

method 4) adding a good solvent into the apraxitant to dissolve the apraxitant at room temperature, adding water to force crystallization, and then separating and drying the precipitated crystal to obtain the crystal form III;

the good solvent is selected from one or more of acetone, butanone, acetonitrile and propionitrile;

the present invention also provides, according to the object of the present invention, crystalline form IV of apraxitant (hereinafter referred to as "crystalline form IV").

The form IV has an X-ray powder diffraction with characteristic peaks at the following 2 θ angular positions using Cu-ka radiation: 4.5 +/-0.2 degrees, 6.6 +/-0.2 degrees, 13.5 +/-0.2 degrees, 27.4 +/-0.2 degrees and 30.1 +/-0.2 degrees.

The form IV also has characteristic peaks at the following 2 θ angular positions: 7.0 +/-0.2 degrees, 10.8 +/-0.2 degrees, 17.0 +/-0.2 degrees, 20.4 +/-0.2 degrees, 21.4 +/-0.2 degrees and 21.8 +/-0.2 degrees.

The form IV also has characteristic peaks at the following 2 θ angular positions: 15.1 +/-0.2 degrees, 17.7 +/-0.2 degrees, 20.0 +/-0.2 degrees, 22.4 +/-0.2 degrees, 23.9 +/-0.2 degrees, 24.4 +/-0.2 degrees, 25.0 +/-0.2 degrees, 26.4 +/-0.2 degrees, 26.7 +/-0.2 degrees, 32.8 +/-0.2 degrees, 34.7 +/-0.2 degrees and 36.0 +/-0.2 degrees.

According to the purpose of the invention, the invention also provides a preparation method of crystalline form IV of apraxitant, which is prepared by adopting any one of the following two methods:

method 1) adding N, N-dimethylformamide into apraxitant to dissolve the apraxitant, adding a poor solvent at a high temperature to keep the system still dissolved, cooling and crystallizing, and separating and drying the precipitated crystal to obtain the crystal form IV;

the elevated temperature is selected from the range of 40 ℃ to the boiling point of the solvent system; the temperature after cooling is selected from 0-30 ℃, and room temperature is preferred;

the volume ratio of the poor solvent to the N, N-dimethylformamide is 0.1-100: 1, preferably 0.5-50: 1, and more preferably 0.5-20: 1;

method 2) adding a pre-mixed N, N-dimethylformamide and poor solvent mixed solution into the apraxitant, dissolving the system at a high temperature, cooling and crystallizing, separating and drying the precipitated crystal to obtain the crystal form IV;

the present invention also provides crystalline form V of apraxitant (hereinafter referred to as "form V"), which is an object of the present invention.

The X-ray powder diffraction of the crystal form V has characteristic peaks at the following 2 theta angle positions by using Cu-K alpha radiation: 12.0 +/-0.2 degrees, 13.0 +/-0.2 degrees, 17.1 +/-0.2 degrees, 23.9 +/-0.2 degrees and 24.3 +/-0.2 degrees.

The form V also has characteristic peaks at the following 2 θ angular positions: 14.7 +/-0.2 degrees, 20.0 +/-0.2 degrees, 21.1 +/-0.2 degrees, 25.9 +/-0.2 degrees, 26.7 +/-0.2 degrees and 35.1 +/-0.2 degrees.

The form V also has characteristic peaks at the following 2 θ angular positions: 16.6 +/-0.2 degrees, 19.8 +/-0.2 degrees, 22.9 +/-0.2 degrees, 26.2 +/-0.2 degrees, 27.1 +/-0.2 degrees, 27.7 +/-0.2 degrees, 29.3 +/-0.2 degrees, 30.0 +/-0.2 degrees, 31.0 +/-0.2 degrees, 32.7 +/-0.2 degrees and 35.4 +/-0.2 degrees.

According to an object of the present invention, the present invention also provides a process for the preparation of crystalline form V of apraxitant:

adding a good solvent into the apraxitant to dissolve the apraxitant at high temperature, adding cyclohexane at high temperature to keep the system still dissolved, cooling and crystallizing, and separating and drying the precipitated crystal to obtain the crystal form V;

the good solvent is selected from C3-C6 ester, and the good solvent can be selected from one ester or a mixture of esters;

the volume ratio of the cyclohexane to the good solvent is 0.1-100: 1, preferably 0.5-50: 1, and more preferably 0.5-20: 1;

the present invention also provides, according to the object of the present invention, crystalline form VI of apraxitant (hereinafter referred to as "crystalline form VI").

The X-ray powder diffraction of said crystalline form VI has characteristic peaks at the following 2 θ angular positions using Cu-ka radiation: 6.3 +/-0.2 degrees, 13.3 +/-0.2 degrees, 16.5 +/-0.2 degrees and 21.1 +/-0.2 degrees.

The form VI also has characteristic peaks at the following 2 θ angular positions: 10.3 +/-0.2 degrees, 14.5 +/-0.2 degrees, 24.3 +/-0.2 degrees, 24.9 +/-0.2 degrees, 25.8 +/-0.2 degrees, 27.7 +/-0.2 degrees and 28.6 +/-0.2 degrees.

The form VI also has characteristic peaks at the following 2 θ angular positions: 17.1 +/-0.2 degrees, 19.9 +/-0.2 degrees, 21.6 +/-0.2 degrees, 23.7 +/-0.2 degrees, 26.5 +/-0.2 degrees, 26.7 +/-0.2 degrees, 28.2 +/-0.2 degrees, 29.2 +/-0.2 degrees, 29.8 +/-0.2 degrees, 31.8 +/-0.2 degrees, 32.5 +/-0.2 degrees, 35.4 +/-0.2 degrees and 37.4 +/-0.2 degrees.

According to an object of the present invention, the present invention also provides a process for the preparation of crystalline form VI of apraxitant:

adding ethylene glycol dimethyl ether into the apraxitant to dissolve the apraxitant at high temperature, cooling and crystallizing, and separating and drying the precipitated crystal to obtain the crystal form VI;

the elevated temperature is selected from the range of 40 ℃ to the boiling point of the solvent system; the temperature after cooling is selected from-10-30 ℃, preferably-5-5 ℃;

in the preparation methods of the crystal form I, the crystal form II, the crystal form III, the crystal form IV, the crystal form V and the crystal form VI of the present invention, the stirring may be performed by a conventional method in the art, such as magnetic stirring, mechanical stirring, and the like. The stirring rate is 50-1800 rpm, preferably 300-900 rpm.

In the preparation method of crystal form I, crystal form II, crystal form III, crystal form IV, crystal form V and crystal form VI, the precipitated crystal is separated, washed and dried by adopting the conventional method in the field. Said separation is carried out using methods conventional in the art, such as filtration, centrifugation, and the like; the specific operation of filtration is: placing a sample to be separated on filter paper, and performing vacuum filtration; the specific operation of centrifugation is: the sample to be separated is placed in a centrifuge tube and then spun at high speed until all the solids settle to the bottom of the centrifuge tube, for example at a centrifuge rate of 6000 rpm. The washing solvent is preferably the same as the solvent used in the preparation method of the crystal form, and the dosage of the washing solvent is 0.2-10 times of that of the crystallization solvent. Drying is carried out by adopting a conventional method in the field, such as natural drying, forced air drying or reduced pressure drying; the drying equipment is a fume hood, a blast oven or a vacuum oven; the drying may be carried out under reduced pressure or not, preferably under a pressure of less than 0.09MPa, at 30-50 deg.C for 10-72 hours, preferably 10-48 hours, more preferably 10-24 hours.

The invention relates to novel crystal forms of apraxitant, which are disclosed for the first time, and comprise a crystal form I, a crystal form II, a crystal form III, a crystal form IV, a crystal form V and a crystal form VI.

In some embodiments, the above-described novel crystalline forms of the invention are pure, single, and substantially free of any other crystalline forms in admixture. As used herein, "substantially free" when used in reference to a novel form means that the form contains less than 20% by weight of the other form, particularly less than 10% by weight of the other form, more particularly less than 5% by weight of the other form, and even more particularly less than 1% by weight of the other form.

In the present invention, "crystal" or "crystalline form" means that it is confirmed by the shown X-ray diffraction pattern characterization. One skilled in the art will appreciate that the physicochemical properties discussed herein can be characterized with experimental error depending on the conditions of the instrument, sample preparation and purity of the sample. In particular, it is well known to those skilled in the art that the X-ray diffraction pattern will generally vary with the conditions of the instrument. It is particularly noted that the relative intensities of the X-ray diffraction patterns may also vary with the experimental conditions, so that the order of the peak intensities cannot be considered as the sole or determining factor. In addition, experimental errors in peak angles are typically 5% or less, and these angle errors should also be taken into account, typically allowing for errors of ± 0.2. In addition, due to the influence of experimental factors such as sample height, an overall shift in peak angle is caused, and a certain shift is usually allowed. Thus, it will be understood by those skilled in the art that the X-ray diffraction pattern of a crystalline form of the present invention need not be identical to the X-ray diffraction pattern of the examples referred to herein. Any crystalline form having the same or similar pattern as the characteristic peaks in these patterns is within the scope of the present invention. One skilled in the art can compare the profiles listed in the present invention with a profile of an unknown crystalline form to confirm whether the two sets of profiles reflect the same or different crystalline forms.

"crystalline form" and "polymorphic form" and other related terms refer herein to the presence of a solid compound in a particular crystalline state within the crystal structure. The difference of the physicochemical properties of the polymorphism can be reflected in the aspects of storage stability, compressibility, density, dissolution speed and the like. In extreme cases, differences in solubility or dissolution rate can cause drug inefficiency and even toxicity.

Polymorphs of a drug may be obtained by methods including, but not limited to: freeze-drying, melt recrystallization, melt cooling, solvent recrystallization, solvent loss, rapid volatilization, rapid temperature reduction, slow temperature reduction, vapor diffusion, sublimation, suspension and grinding. Polymorphs can be detected, discovered and classified by X-ray powder diffraction (XRPD), Differential Scanning Calorimetry (DSC), thermogravimetric analysis (TGA), optical microscopy, hygroscopicity and the like.

The experimental procedure can be carried out mostly at or near room temperature. Here, the experiment is performed at or near the same temperature as the room or fume hood. Typically this temperature is from 15 ℃ to 25 ℃, alternatively 17 ℃, alternatively 22 ℃. The acquisition of form VI needs to be achieved at lower temperatures and can be operated in a low temperature reactor.

The experimental procedure or procedure may be carried out "overnight". This step is referred to herein as spanning the night period, with no active observation of experimental phenomena during the overnight period. This period may be 8 to 22 hours, or 10 to 18 hours, typically 16 hours.

Unless otherwise specified, the crystalline forms described in the present invention may be subjected to a drying step. Drying may be carried out at room temperature or higher. The crystalline form material may be dried at a temperature of from 20 ℃ to about 60 ℃, alternatively to 40 ℃, alternatively to 50 ℃. The drying time may be 2-48 hours, or overnight. Drying may be carried out in a fume hood, a forced air oven or a vacuum oven.

The crystallization mode adopted in the invention comprises room temperature volatilization, slow volatilization, crystal slurry, macromolecule template volatilization recrystallization, cooling recrystallization and anti-solvent recrystallization.

Room temperature evaporation is, for example, to evaporate a clear solution of a sample in an open 5mL glass vial under specific temperature conditions (typically room temperature). The nitrogen blowing method or direct room temperature volatilization can be used.

Slow evaporation (small pore evaporation) is for example performed by placing the sample clear solution in a perforated 5mL glass vial and slowly evaporating in air.

The slurry is prepared, for example, by stirring a supersaturated solution of the sample (with undissolved solids present) in a different solvent system, typically for a period of 2 hours to 2 weeks.

The cooling recrystallization is, for example, dissolving the sample in a suitable solvent under a specific high temperature condition, placing the solution in a 5mL glass vial, placing the vial in a temperature-changing shaking table, sequentially cooling the solution at a certain cooling rate, and stirring the solution at room temperature overnight. The temperature of the experiment may be 75-0 deg.C, preferably 50-15 deg.C. The sample solutions were incubated for 1 hour to 2 days at each specified temperature.

The anti-solvent recrystallization is, for example, a method in which a sample is dissolved in a good solvent, the solution is dissolved by ultrasonic waves, an appropriate amount of the anti-solvent is added, and the mixture is stirred at room temperature.

The invention solves the problems existing in the prior crystal form by providing six different crystal forms of the apraxitant, and the polymorphism of the apraxitant is beneficial to accurately obtaining the crystal form condition of the apraxitant so as to preferentially select a proper crystal form for drug development, ensure the quality of the drug and obtain better clinical treatment effect.

In addition, the present invention also provides a pharmaceutical composition comprising a therapeutically and/or prophylactically effective amount of one or more of said crystalline form I, crystalline form II, crystalline form III, crystalline form IV, crystalline form V or crystalline form VI of a sulfonamide compound, and at least one pharmaceutically acceptable carrier.

The pharmaceutical compositions of the present invention may be in solid or liquid form. If the pharmaceutical composition is in a liquid state, the six different crystals of crystalline form I-VI of apraxitant above remain as a solid in the pharmaceutical composition, e.g., as a suspension.

The pharmaceutically acceptable carrier of the present invention includes, but is not limited to: diluents such as starch, pregelatinized starch, lactose, powdered cellulose, microcrystalline cellulose, calcium hydrogen phosphate, tricalcium phosphate, mannitol, sorbitol, sugar, and the like; an adhesive; such as gum arabic, guar gum, gelatin, polyvinylpyrrolidone, hydroxypropyl cellulose, hydroxypropyl methylcellulose, polyethylene glycol, and the like; disintegrants, such as starch, sodium starch glycolate, pregelatinized starch, crospovidone, croscarmellose sodium, colloidal silicon dioxide, and the like; lubricants, such as stearic acid, magnesium stearate, zinc stearate, sodium benzoate, sodium acetate, and the like; glidants such as colloidal silicon dioxide and the like; complex-forming agents, such as various grades of cyclodextrins and resins; release rate controlling agents such as hydroxypropyl cellulose, hydroxymethyl cellulose, hydroxypropyl methylcellulose, ethyl cellulose, methyl methacrylate, waxes, and the like. Other pharmaceutically acceptable carriers that may be used include, but are not limited to, film forming agents, plasticizers, colorants, flavoring agents, viscosity modifiers, preservatives, antioxidants, and the like.

The pharmaceutical composition can be prepared into certain dosage forms, preferably oral administration, parenteral administration (including subcutaneous administration, intramuscular administration and intravenous administration), rectal administration, transdermal administration, nasal administration and other dosage forms, including but not limited to solid dosage forms, liquid dosage forms, semi-liquid dosage forms, aerosol or suppository and the like. For example, dosage forms suitable for oral administration include tablets, capsules, granules, powders, pills, powders, lozenges, syrups or suspensions; dosage forms suitable for parenteral administration include aqueous or non-aqueous solutions or emulsions; dosage forms suitable for rectal administration include suppositories with hydrophilic or hydrophobic carriers; dosage forms suitable for transdermal administration include ointments, creams; dosage forms suitable for nasal administration include aerosols, sprays. The above dosage forms may be adapted for rapid, delayed or modified release of the active ingredient, as desired. Preferably, the pharmaceutical composition is an oral preparation or an injection preparation; more preferably, the pharmaceutical composition is a solid oral preparation, including tablets, capsules, granules, pills and powders.

The pharmaceutical compositions may be prepared using methods well known to those skilled in the art. In preparation, the crystalline form I-VI of apraxitant of the invention, or combinations thereof, is mixed with one or more pharmaceutically acceptable carriers, optionally with one or more other active ingredients. The solid preparation can be prepared by mixing, granulating and the like.

According to the purpose of the invention, the invention provides the usage of the crystal of the crystalline form I-VI of the apraxitant and the pharmaceutical composition in preparing medicines for treating diseases caused by endothelin increase and related to vasoconstriction, cell proliferation and inflammation, such as hypertension, pulmonary hypertension, coronary heart disease, heart failure, renal and myocardial atrophy, renal failure, cerebral ischemia, cerebral vasospasm, dementia, migraine, subarachnoid hemorrhage, Rena's syndrome, portal hypertension, arteriosclerosis, restenosis after angioplasty, cancer, asthma and the like.

Drawings

Figure 1 is an X-ray powder diffraction pattern of form I of the present invention.

Figure 2 is an X-ray powder diffraction pattern of form II of the present invention.

Figure 3 is an X-ray powder diffraction pattern of form III of the present invention.

Figure 4 is an X-ray powder diffraction pattern of form IV of the present invention.

Figure 5 is an X-ray powder diffraction pattern of form V of the present invention.

Figure 6 is an X-ray powder diffraction pattern of form VI of the present invention.

Figure 7 is a DSC diagram of form I of the present invention.

Figure 8 is a DSC diagram of form II of the present invention.

Figure 9 is a DSC diagram of form III of the present invention.

Figure 10 is a DSC diagram of form IV of the present invention.

Figure 11 is a DSC diagram of form V of the present invention.

Fig. 12 is an X-ray powder diffraction pattern of comparative example 1.

FIG. 13 is a DSC chart of comparative example 1.

Fig. 14 is an X-ray powder diffraction pattern of comparative example 2.

FIG. 15 is a DSC chart of comparative example 2.

Fig. 16 is an X-ray powder diffraction pattern of comparative example 3.

FIG. 17 is a DSC chart of comparative example 3.

Detailed Description

The invention is further defined by reference to the following examples describing in detail the methods of making and using the crystalline forms of the invention. It will be apparent to those skilled in the art that many modifications, both to materials and methods, may be practiced without departing from the scope of the invention.

The instrument and method for data acquisition:

high Performance Liquid Chromatography (HPLC) is SHIMADZU LC-15C, chromatographic column is Agilent SB-C184.6 × 250mm 5um, flow rate is 1.0ml/min, column temperature is 30 deg.C, wavelength is 205nm, mobile phase A is 0.1% phosphoric acid and acetonitrile is 90: 10, phase B is acetonitrile, gradient elution time is 30 min.

The NMR instrument is Bruker DRX-400FT (Germany), 1HNMR in DMSO-d₆Chemical shifts were measured in ppm based on Tetramethylsilane (TMS).

The mass spectrometer was a Thermo-Scientific active spectrometer and was measured using ESI mode.

The apparatus used for X-ray powder diffraction (XPRD) was Read before inserting MiniFlex 300/600 using Ka X-rays with a copper target wavelength of 1.54nm, a theta-2 theta goniometer, a Mo monochromator, a Lynxeye detector under operating conditions of 40kV and 40 mA. The instrument is calibrated prior to use with a standard (typically corundum) carried by the instrument itself. The collection software was MiniFlex guidelines and the analysis software was origin 75. The samples were tested at room temperature and the sample to be tested was placed on an organic glass slide. The detailed detection conditions were as follows: angle range: 3-40 or 50 ° 2 θ; step length: 0.02 ° 2 θ; speed: 0.2 s/step. Unless otherwise specified, the samples were not ground prior to testing.

Differential Thermal analysis (DSC) data were obtained from METTLER TOLEDO DSC3, instrument control software is Thermal Advantage, and analytical software is STARE Default DB V15.00. Typically, 1-10 mg of the sample was placed in an uncapped (unless otherwise specified) aluminum crucible and the sample was raised from room temperature to 250 ℃ at a ramp rate of 10 ℃/min under the protection of 50mL/min dry N2, while the TA software recorded the heat change of the sample during the ramp. In the present application, melting point is reported in terms of starting temperature.

Preparation example: preparation of apraxitant

Prepared according to the method described in example 3 variant I of patent document EP2907811, and the white solid is obtained by column chromatography, with an HPLC purity of 98.1%; the 1HNMR data are as follows:¹H-NMR(400MHz，DMSO-d6)δppm 9.79(s，1H)，8.81(s，2H)，8.48(s，1H) 7.54-7.52(d, 2H), 7.18(s br, 2H), 7.15-7.13(d, 2H), 4.66-4.62(m, 2H), 4.58-4.55(m, 2H); theoretical molecular weight: 545(M + H), measured molecular weight: 545(M + H), 567(M + Na).

Example 1

Putting 10.0mg of apraxitant into a 5mL glass vial, adding 0.5mL of methyl acetate, heating to dissolve the apraxitant, stirring for 4 days at room temperature, centrifuging, and performing vacuum drying for 16 hours at 40 ℃ to obtain the crystal form I. The yield is 7.8 mg; the yield was 78%. The X-ray powder diffraction pattern is shown in FIG. 1. The DSC spectrum is shown in figure 7, and the DSC shows that: melting point of form I is 152.5 ℃.

Example 2

Putting 10.0mg of apraxitant into a 5mL glass vial, adding 0.5mL of acetone, heating to dissolve the apraxitant, stirring for 3 days at room temperature, centrifuging, and performing vacuum drying for 16 hours at 40 ℃ to obtain the crystal form I. The yield was 7.6 mg; the yield was 76%. The X-ray powder diffraction pattern is substantially the same as in fig. 1.

Example 3

Putting 10.0mg of apraxitant into a 5mL glass vial, adding 0.5mL of chloroform mixed solution, heating to dissolve the solution, stirring at room temperature for 4 days, centrifuging, and performing vacuum drying at 40 ℃ for 16 hours to obtain the crystal form I. The yield is 8.6 mg; the yield was 86%. The X-ray powder diffraction pattern is substantially the same as in fig. 1.

Example 4

Putting 10.0mg of apraxitant into a 5mL glass vial, adding 0.4mL of tetrahydrofuran, heating to dissolve the apraxitant, stirring at room temperature for 7 days, centrifuging, and performing vacuum drying at 40 ℃ for 16 hours to obtain the crystal form I. The yield was 7.6 mg; the yield was 76%. The X-ray powder diffraction pattern is substantially the same as in fig. 1.

Example 5

Placing 10.0mg of apraxitant in a 5mL glass vial, adding 0.3mL of an ethyl acetate/isopropyl acetate (v/v is 1: 1) mixed solution, heating to dissolve the solution, stirring at room temperature for 14 days, centrifuging, and performing vacuum drying at 40 ℃ for 16 hours to obtain the crystal form I. The yield is 6.6 mg; the yield was 66%. The X-ray powder diffraction pattern is substantially the same as in fig. 1.

Example 6

Putting 10.0mg of apraxitant into a 5mL glass vial, adding 0.3mL of acetone and 0.1mL of methyl tetrahydrofuran, heating to dissolve, stirring at room temperature for 7 days, centrifuging, and performing vacuum drying at 40 ℃ for 16 hours to obtain the crystal form I. The yield is 8.1 mg; the yield was 81%. The X-ray powder diffraction pattern is substantially the same as in fig. 1.

Example 7

Placing 10.0mg of apraxitant in a 5mL glass vial, adding 0.2mL of chloroform and 0.2mL of ethyl acetate, heating to dissolve, stirring at room temperature for 10 days, centrifuging, and performing vacuum drying at 40 ℃ for 16 hours to obtain the crystal form I. The yield was 7.7 mg; the yield was 77%. The X-ray powder diffraction pattern is substantially the same as in fig. 1.

Example 8

Putting 10.0mg of apraxitant into a 5mL glass vial, adding 0.1mL of acetone and 0.2mL of ethyl acetate, heating to dissolve, stirring at room temperature for 8 days, centrifuging, and performing vacuum drying at 40 ℃ for 16 hours to obtain the crystal form I. The yield was 7.5 mg; the yield was 75%. The X-ray powder diffraction pattern is substantially the same as in fig. 1.

Example 9

Placing 10.0mg of apraxitant in a 5mL glass vial, adding 0.2mL of ethyl acetate, heating to dissolve the mixture, adding 0.5mL of n-heptane, stirring at room temperature for 7 days, centrifuging, and performing vacuum drying at 40 ℃ for 16 hours to obtain the crystal form I. The yield is 8.5 mg; the yield was 85%. The X-ray powder diffraction pattern is substantially the same as in fig. 1.

Example 10

Putting 10.0mg of apraxitant into a 5mL glass vial, adding 0.2mL of acetone, heating to dissolve the mixture, adding 0.2mL of n-hexane, stirring at room temperature for 7 days, centrifuging, and performing vacuum drying at 40 ℃ for 16 hours to obtain the crystal form I. The yield was 9.2 mg; the yield was 92%. The X-ray powder diffraction pattern is substantially the same as in fig. 1.

Example 11

Placing 10.0mg of apraxitant in a 5mL glass vial, adding 0.2mL of acetonitrile, heating to dissolve, adding 0.3mL of n-heptane, stirring at room temperature for 7 days, centrifuging, and performing vacuum drying at 40 ℃ for 16 hours to obtain the crystal form I. The yield is 8.2 mg; the yield was 82%. The X-ray powder diffraction pattern is substantially the same as in fig. 1.

Example 12

Placing 10.0mg of apraxitant into a 5mL glass vial, adding 0.7mL of acetonitrile/toluene (v/v is 1: 5) mixed solution, heating to dissolve, stirring at room temperature for 14 days, centrifuging, and performing vacuum drying at 40 ℃ for 16 hours to obtain the crystal form I. The yield is 6.8 mg; the yield was 68%. The X-ray powder diffraction pattern is substantially the same as in fig. 1.

Example 13

Placing 10.0mg of apraxitant in a 5mL glass vial, adding 0.2mL of acetone, heating to dissolve, adding 0.5mL of toluene, stirring at room temperature for 12 days, centrifuging, and performing vacuum drying at 40 ℃ for 16 hours to obtain the crystal form I. The yield is 8.0 mg; the yield was 80%. The X-ray powder diffraction pattern is substantially the same as in fig. 1.

Example 14

Placing 10.0mg of apraxitant into a 5mL glass vial, adding 0.6mL of ethyl acetate/toluene (v/v is 1: 3) mixed solution, heating to dissolve, stirring at room temperature for 14 days, centrifuging, and performing vacuum drying at 40 ℃ for 16 hours to obtain the crystal form I. The yield is 6.8 mg; the yield was 68%. The X-ray powder diffraction pattern is substantially the same as in fig. 1.

Example 15

Putting 10.0mg of apraxitant into a 5mL glass vial, adding 0.1mL of DMSO (dimethyl sulfoxide) for dissolving, adding 3mL of water, stirring at room temperature for 1 hour, centrifuging, and performing vacuum drying at 60 ℃ for 16 hours to obtain the crystal form I. The yield was 9.1 mg; the yield was 91%. The X-ray powder diffraction pattern is substantially the same as in fig. 1.

Example 16

Putting 10.0mg of apraxitant into a 5mL glass vial, adding 0.1mL of DMF for dissolving, adding 2mL of water, stirring at room temperature for 1 hour, centrifuging, and performing vacuum drying at 60 ℃ for 16 hours to obtain the crystal form I. The yield is 8.8 mg; the yield was 88%. The X-ray powder diffraction pattern is substantially the same as in fig. 1.

Example 17

And (2) putting 10.0mg of apraxitant into a 5mL glass vial, adding 0.3mL of dichloromethane, heating to dissolve the mixture, adding 0.4mL of n-heptane, stirring at room temperature for 2 days, centrifuging, and performing vacuum drying at 40 ℃ for 16 hours to obtain the crystal form II. The yield is 8.8 mg; the yield was 88%. The X-ray powder diffraction pattern is shown in figure 2. The DSC profile is shown in figure 8 and shows: the melting point of form II is 108.1 ℃.

Example 18

And (3) putting 10.0mg of apraxitant into a 5mL glass vial, adding 0.3mL of dichloromethane, heating to dissolve the mixture, adding 0.3mL of cyclohexane, stirring at room temperature for 2 days, centrifuging, and performing vacuum drying at 40 ℃ for 16 hours to obtain the crystal form II. The yield is 8.3 mg; the yield was 83%. The X-ray powder diffraction pattern is substantially the same as that of fig. 2.

Example 19

And (3) putting 10.0mg of apraxitant into a 5mL glass vial, adding 0.3mL of dichloromethane, heating to dissolve the mixture, adding 0.5mL of cyclohexane, stirring at room temperature for 2 days, centrifuging, and performing vacuum drying at 40 ℃ for 16 hours to obtain the crystal form II. The yield is 8.7 mg; the yield was 87%. The X-ray powder diffraction pattern is substantially the same as that of fig. 2.

Example 20

Putting 10.0mg of apraxitant into a 5mL glass vial, adding 0.3mL of dichloromethane, heating, dissolving, adding 0.3mL of n-hexane, stirring at room temperature for 2 days, centrifuging, and performing vacuum drying at 40 ℃ for 16 hours to obtain the crystal form II. The yield is 8.9 mg; the yield was 89%. The X-ray powder diffraction pattern is substantially the same as that of fig. 2.

Example 21

Placing 10.0mg of apraxitant in a 5mL glass vial, adding 0.3mL of tetrahydrofuran, heating to dissolve, adding 0.4mL of n-heptane, stirring for 5 days at room temperature, centrifuging, and performing vacuum drying at 40 ℃ for 16 hours to obtain the crystal form III. The yield is 7.8 mg; the yield was 78%. The X-ray powder diffraction pattern is shown in figure 3. The DSC profile is shown in figure 9, showing: melting point of form III is 148.1 ℃.

Example 22

Placing 10.0mg of apraxitant in a 5mL glass vial, adding 0.3mL of ethylene glycol dimethyl ether, heating to dissolve the mixture, adding 1mL of toluene, stirring at room temperature for 16 days, centrifuging, and performing vacuum drying at 40 ℃ for 16 hours to obtain the crystal form III. The yield is 8.9 mg; the yield was 89%. The X-ray powder diffraction pattern substantially corresponds to that of figure 3.

Example 23

Placing 10.0mg of apraxitant into a 5mL glass vial, adding 0.5mL of a methyl tetrahydrofuran/toluene (v/v is 1: 5) mixed solution, heating to dissolve the solution, stirring at room temperature for 13 days, centrifuging, and performing vacuum drying at 40 ℃ for 16 hours to obtain the crystal form III. The yield was 7.9 mg; the yield was 79%. The X-ray powder diffraction pattern substantially corresponds to that of figure 3.

Example 24

Putting 10.0mg of apraxitant into a 5mL glass vial, adding 0.3mL of dioxane solvent, adding 0.5mL of n-hexane, stirring at room temperature for 16 days, centrifuging, and performing vacuum drying at 40 ℃ for 16 hours to obtain the crystal form III. The yield was 6.9 mg; the yield was 69%. The X-ray powder diffraction pattern substantially corresponds to that of figure 3.

Example 25

Putting 10.0mg of apraxitant into a 5mL glass vial, adding 0.2mL of methyltetrahydrofuran, heating to dissolve, adding 1.5mL of n-hexane, stirring at room temperature for 3 hours, centrifuging, and performing vacuum drying at 40 ℃ for 16 hours to obtain the crystal form III. The yield was 9.5 mg; the yield was 95%. The X-ray powder diffraction pattern substantially corresponds to that of figure 3.

Example 26

Putting 10.0mg of apraxitant into a 5mL glass vial, adding 0.3mL of tetrahydrofuran, heating to dissolve, adding 1.5mL of cyclohexane, stirring at room temperature for 3 hours, centrifuging, and performing vacuum drying at 40 ℃ for 16 hours to obtain the crystal form III. The yield is 8.3 mg; the yield was 83%. The X-ray powder diffraction pattern substantially corresponds to that of figure 3.

Example 27

And (3) putting 10.0mg of apraxitant into a 5mL glass vial, adding 0.2mL of butanone, heating to dissolve the mixture, adding 0.8mL of water, stirring at room temperature for 3 hours, centrifuging, and performing vacuum drying at 60 ℃ for 16 hours to obtain the crystal form III. The yield is 8.0 mg; the yield was 80%. The X-ray powder diffraction pattern substantially corresponds to that of figure 3.

Example 28

And (3) putting 10.0mg of apraxitant into a 5mL glass vial, adding 0.2mL of acetonitrile, heating to dissolve the mixture, adding 1.5mL of water, stirring at room temperature for 1 hour, centrifuging, and performing vacuum drying at 60 ℃ for 16 hours to obtain the crystal form III. The yield was 9.0 mg; the yield was 90%. The X-ray powder diffraction pattern substantially corresponds to that of figure 3.

Example 29

And (3) putting 10.0mg of apraxitant into a 5mL glass vial, adding 0.2mL of acetone, heating to dissolve, adding 1.5mL of water, stirring at room temperature for 1 hour, centrifuging, and performing vacuum drying at 60 ℃ for 16 hours to obtain the crystal form III. The yield was 9.1 mg; the yield was 91%. The X-ray powder diffraction pattern substantially corresponds to that of figure 3.

Example 30

Putting 10.0mg of apraxitant into a 5mL glass vial, adding 0.2mL of propionitrile, heating to dissolve the mixture, adding 1.5mL of water, stirring at room temperature for 1 hour, centrifuging, and performing vacuum drying at 60 ℃ for 16 hours to obtain the crystal form III. The yield is 8.8 mg; the yield was 88%. The X-ray powder diffraction pattern substantially corresponds to that of figure 3.

Example 31

Placing 10.0mg of apraxitant in a 5mL glass vial, adding 0.2mL of DMF, heating to dissolve, adding 0.3mL of toluene, stirring at room temperature for 15 days, centrifuging, and performing vacuum drying at 40 ℃ for 16 hours to obtain the crystal form IV. The yield is 6.8 mg; the yield was 68%. The X-ray powder diffraction pattern is shown in figure 4. The DSC profile is shown in figure 10 and shows: form IV melting point is 167.1 ℃.

Example 32

Putting 10.0mg of apraxitant into a 5mL glass vial, adding 0.2mL of DMF, heating to dissolve, adding 0.4mL of cyclohexane, stirring at room temperature for 15 days, centrifuging, and performing vacuum drying at 40 ℃ for 16 hours to obtain the crystal form IV. The yield was 7.5 mg; the yield was 75%. The X-ray powder diffraction pattern is shown in figure 4. The DSC profile is essentially the same as in figure 4.

Example 33

Putting 10.0mg of apraxitant into a 5mL glass vial, adding 0.2mL of ethyl acetate, heating to dissolve, adding 0.3mL of cyclohexane, stirring at room temperature for 7 days, centrifuging, and performing vacuum drying at 40 ℃ for 16 hours to obtain the crystal form V. The yield is 6.8 mg; the yield was 68%. The X-ray powder diffraction pattern is shown in figure 5. The DSC profile is shown in figure 11, which shows: melting point of form V is 143.7 ℃.

Example 34

Putting 10.0mg of apraxitant into a 5mL glass vial, adding 0.2mL of isopropyl acetate, heating to dissolve, adding 0.3mL of cyclohexane, stirring at room temperature for 7 days, centrifuging, and vacuum-drying at 40 ℃ for 16 hours to obtain the crystal form V. The yield is 6.5 mg; the yield was 68%. The X-ray powder diffraction pattern was substantially the same as that of fig. 5.

Example 35

Putting 10.0mg of apraxitant into a 5mL glass vial, adding 0.2mL of methyl acetate, heating to dissolve the mixture, adding 0.3mL of cyclohexane, stirring at room temperature for 7 days, centrifuging, and performing vacuum drying at 40 ℃ for 16 hours to obtain the crystal form V. The yield was 6.9 mg; the yield was 69%. The X-ray powder diffraction pattern was substantially the same as that of fig. 5.

Example 36

Placing 10.0mg of apraxitant in a 5mL glass vial, adding 0.3mL of ethylene glycol dimethyl ether, heating to dissolve, stirring at room temperature for 5 days, cooling to 0 ℃, continuing to stir for 3 days, centrifuging, and vacuum-drying at 60 ℃ for 16 hours to obtain the crystal form VI. The yield was 5.9 mg; the yield was 59%. The X-ray powder diffraction pattern is shown in figure 6.

Example 37

Placing 10.0mg of apraxitant in a 5mL glass vial, adding 0.5mL of ethylene glycol dimethyl ether, heating to dissolve, stirring at room temperature for 5 days, cooling to 0 ℃, continuing to stir for 5 days, centrifuging, and vacuum-drying at 60 ℃ for 16 hours to obtain the crystal form VI. The yield is 4.8 mg; the yield was 48%. The X-ray powder diffraction pattern was substantially the same as that of fig. 6.

Comparative example 1

With reference to the crystallization process described in example 1.iv of patent WO2009024906, apraxitant was crystallized using dichloromethane as crystallization solvent. The specific operation is as follows: putting 1g of apraxitant into a 10mL reaction bottle, adding 3mL of dichloromethane, heating to dissolve, slowly cooling to crystallize, stirring overnight at 0-5 ℃, filtering, and vacuum-drying at 40 ℃ for 16 hours to obtain 0.55g of white crystalline solid with the yield of 55%. The X-ray powder diffraction pattern is shown in figure 12, and the DSC pattern is shown in figure 13. XRD and DSC results show that: the crystalline form obtained in comparative example 1 is not a pure single crystalline form.

Comparative example 2

With reference to the process described in example 3, variant II of example 3 of patent EP2907811, crystalline apraxitant is crystallized with dichloromethane as the crystallization solvent and the crystalline form III obtained in example 21 is added to induce crystallization, in the following manner: putting 1g of apraxitant into a 10mL reaction bottle, adding 5mL of dichloromethane, heating to dissolve, slowly cooling to room temperature, adding a small amount of the crystal form III obtained in example 21 for induced crystallization, then continuously cooling to 4 ℃, stirring for 1h, filtering, leaching with cold dichloromethane, and vacuum drying at 40 ℃ for 16 h to obtain 0.65g of white crystalline solid with the yield of 65%. The X-ray powder diffraction pattern is shown in figure 14, and the DSC pattern is shown in figure 15. XRD and DSC results show that: the crystalline form obtained in comparative example 2 is not a pure single crystalline form.

Comparative example 3

With reference to the process described in example 3, variant I of patent EP2907811, apraxitant is crystallized from ethyl acetate as crystallization solvent, in the following manner: putting 1g of apraxitant into a 10mL reaction bottle, adding 2mL of ethyl acetate, heating to dissolve the apraxitant, slowly cooling to crystallize, adding 2mL of ethyl acetate after obvious crystal precipitation to cool to room temperature, filtering, and drying in vacuum at 40 ℃ for 16 hours to obtain 0.37g of white crystalline solid with the yield of 37%. The X-ray powder diffraction pattern is shown in figure 16, and the DSC pattern is shown in figure 17. XRD and DSC results show that: the crystalline form obtained in comparative example 3 is not a pure single crystalline form.

Comparative example 4

With reference to the process described in example 3, variant VI of patent EP2907811, in a mixed solvent of ethyl acetate/isopropyl acetate (V/V-1/1), apraxitant is crystallized by acidification with slow addition of 1M sulfuric acid, in the following manner: 1g of apraxitant is put into a 25mL reaction bottle, 8mL of a mixed solvent of ethyl acetate/isopropyl acetate (V/V-1/1) is added, the solution is stirred to be clear, 1.35mL of a 1M sulfuric acid aqueous solution is slowly dripped to carry out acidification crystallization, the solution is washed twice by water (10mL of 2), the solution is slurried in 10mL of water at room temperature for 3h, and after filtration, vacuum drying is carried out at 40 ℃ for 16 h, so that 0.65g of white crystalline solid is obtained, and the yield is 65%. TLC and HPLC monitoring showed: the crystallization solution of comparative example 4 and the product obtained by crystallization contain an acid-degrading impurity of slightly less polarity than apraxitant, and the content is at least more than 10%.

It will be appreciated by those skilled in the art that modifications or variations may be made to the present invention in light of the above teachings. Such modifications and variations are intended to be included herein within the scope of this disclosure and the appended claims.

Claims

1. A crystal form III of apraxitant with a structural formula shown as the following,

characterized in that, using Cu-Kalpha radiation, the Apricoxitan has characteristic peaks in the following 2 theta angle positions in X-ray powder diffraction: 7.6 +/-0.2 degrees, 19.4 +/-0.2 degrees, 21.8 +/-0.2 degrees and 24.3 +/-0.2 degrees;

the apraxitant also has characteristic peaks at the following 2 theta angular positions: 9.5 +/-0.2 degrees, 15.6 +/-0.2 degrees, 17.1 +/-0.2 degrees, 23.4 +/-0.2 degrees, 26.6 +/-0.2 degrees and 28.4 +/-0.2 degrees;

the apraxitant also has characteristic peaks at the following 2 theta angular positions: 18.5 +/-0.2 degrees, 20.0 +/-0.2 degrees, 20.5 +/-0.2 degrees, 22.3 +/-0.2 degrees, 22.8 +/-0.2 degrees, 23.1 +/-0.2 degrees, 25.2 +/-0.2 degrees, 25.6 +/-0.2 degrees, 29.1 +/-0.2 degrees, 31.4 +/-0.2 degrees, 32.7 +/-0.2 degrees and 39.7 +/-0.2 degrees.

2. A crystalline form IV of apraxitant having the structural formula shown below,

characterized in that, using Cu-Kalpha radiation, the Apricoxitan has characteristic peaks in the following 2 theta angle positions in X-ray powder diffraction: 4.5 +/-0.2 degrees, 6.6 +/-0.2 degrees, 13.5 +/-0.2 degrees, 27.4 +/-0.2 degrees and 30.1 +/-0.2 degrees;

the apraxitant also has characteristic peaks at the following 2 theta angular positions: 7.0 +/-0.2 degrees, 10.8 +/-0.2 degrees, 17.0 +/-0.2 degrees, 20.4 +/-0.2 degrees, 21.4 +/-0.2 degrees and 21.8 +/-0.2 degrees;

the apraxitant also has characteristic peaks at the following 2 theta angular positions: 15.1 +/-0.2 degrees, 17.7 +/-0.2 degrees, 20.0 +/-0.2 degrees, 22.4 +/-0.2 degrees, 23.9 +/-0.2 degrees, 24.4 +/-0.2 degrees, 25.0 +/-0.2 degrees, 26.4 +/-0.2 degrees, 26.7 +/-0.2 degrees, 32.8 +/-0.2 degrees, 34.7 +/-0.2 degrees and 36.0 +/-0.2 degrees.

3. A crystal form V of apraxitant with a structural formula shown as the following,

characterized in that, using Cu-Kalpha radiation, the Apricoxitan has characteristic peaks in the following 2 theta angle positions in X-ray powder diffraction: 12.0 +/-0.2 degrees, 13.0 +/-0.2 degrees, 17.1 +/-0.2 degrees, 23.9 +/-0.2 degrees and 24.3 +/-0.2 degrees;

the apraxitant also has characteristic peaks at the following 2 theta angular positions: 14.7 +/-0.2 degrees, 20.0 +/-0.2 degrees, 21.1 +/-0.2 degrees, 25.9 +/-0.2 degrees, 26.7 +/-0.2 degrees and 35.1 +/-0.2 degrees;

the apraxitant also has characteristic peaks at the following 2 theta angular positions: 16.6 +/-0.2 degrees, 19.8 +/-0.2 degrees, 22.9 +/-0.2 degrees, 26.2 +/-0.2 degrees, 27.1 +/-0.2 degrees, 27.7 +/-0.2 degrees, 29.3 +/-0.2 degrees, 30.0 +/-0.2 degrees, 31.0 +/-0.2 degrees, 32.7 +/-0.2 degrees and 35.4 +/-0.2 degrees.

4. A preparation method of crystalline form I of apraxitant is characterized by adopting any one of four methods:

the volume ratio of the water to the good solvent is 0.1-100: 1, preferably 0.5-50: 1, and more preferably 0.5-20: 1.

5. A preparation method of crystalline form III of apraxitant is characterized by adopting any one of 4 methods:

the poor solvent is selected from C5 ═ C8 alkane, and n-hexane, cyclohexane and n-heptane are preferred;

6. A preparation method of crystalline form IV of apraxitant is characterized by adopting any one of two methods:

the volume ratio of the poor solvent to the N, N-dimethylformamide is 0.1-100: 1, preferably 0.5-50: 1, and more preferably 0.5-20: 1.

7. A pharmaceutical composition comprising a therapeutically and/or prophylactically effective amount of one or more of crystalline form III, crystalline form IV, or crystalline form V of apraxitant according to any one of claims 1-3 and at least one pharmaceutically acceptable carrier.

8. The pharmaceutical composition of claim 7, wherein the pharmaceutical composition is an oral formulation or an injectable formulation.

9. The pharmaceutical composition according to claim 8, wherein the pharmaceutical composition is selected from solid oral formulations of tablets, capsules, granules, pills or powders.

10. Use of crystalline form III, crystalline form IV or crystalline form V of apraxitant according to any one of claims 1 to 3 or a pharmaceutical composition according to any one of claims 7 to 9 for the manufacture of a medicament for the treatment of hypertension, pulmonary hypertension, coronary heart disease, heart failure, renal and myocardial atrophy, renal failure, cerebral ischemia, cerebral vasospasm, dementia, migraine, subarachnoid hemorrhage, raney's syndrome, portal hypertension, atherosclerosis, restenosis following angioplasty, cancer, asthma.