阅读说明：本技术 甲磺酸卡莫司他及其溶剂化物的晶型和它们的制备方法和用途 (Crystal forms of camostat mesylate and solvates thereof, and preparation methods and applications of crystal forms and solvates thereof ) 是由 穆帅 李树军 雷永胜 陈蔚 闫少杰 王浩 张慕军 周学福 潘毅 陈华 刘福景 于 2020-05-18 设计创作，主要内容包括：本发明提供一种甲磺酸卡莫司他的晶型、其水合物晶型、其丙酮溶剂化物晶型和其N,N-二甲基甲酰胺溶剂化晶型,以及它们的制备方法和应用。其中,所述甲磺酸卡莫司他的晶型及其水合物晶型具有良好的药用价值。因此,本发明还提供了所述晶型或所述水合物晶型在制备治疗胰腺炎、反流性食管炎和由冠状病毒感染引发的疾病的药物中的应用。(The invention provides a crystal form of camostat mesylate, a hydrate crystal form of camostat mesylate, an acetone solvate crystal form of camostat mesylate, an N, N-dimethylformamide solvate crystal form of camostat mesylate, and preparation methods and applications of the crystal form and the hydrate crystal form of camostat mesylate. Wherein, the crystal form of the camostat mesylate and the hydrate crystal form thereof have good medicinal value. Therefore, the invention also provides the application of the crystal form or the hydrate crystal form in preparing a medicament for treating pancreatitis, reflux esophagitis and diseases caused by coronavirus infection.)

1. A crystalline form of [2- (dimethylamino) -2-oxoethyl ] 4- (4-guanidinobenzoyloxy) phenylacetate mesylate having a chemical structure according to formula (I):

characterized in that the X-ray powder diffraction using Cu-K alpha radiation, expressed in 2 theta + -0.2 angle, has diffraction peaks at 9.52, 12.16, 16.43, 17.10, 19.91, 20.40, 20.68 and 21.60 with an error of + -0.2.

2. The crystalline form of claim 1 having an X-ray powder diffraction pattern with the following characteristic diffraction angles, 2 Θ, interplanar spacings, d, and relative intensities, with an error in 2 Θ of 0.2:

preferably, the crystalline form has an X-ray powder diffraction pattern as shown in figure 3.

3. A hydrate crystal form of 4- (4-guanidinobenzoyloxy) phenylacetic acid [2- (dimethylamino) -2-oxoethyl ] ester methanesulfonate shown as a formula (I),

characterized in that the X-ray powder diffraction expressed in 2 theta angle using Cu-K alpha radiation has diffraction peaks with an error of + -0.2 at 9.56, 12.14, 16.44, 17.10, 19.92, 20.39, 20.67 and 21.60.

4. The hydrate crystalline form according to claim 3, having an X-ray powder diffraction pattern with the following characteristic diffraction angles, 2 θ, interplanar spacings, d, and relative intensities, wherein the error in 2 θ is 0.2:

preferably, the X-ray powder diffraction pattern of the hydrate crystalline form is shown in fig. 6.

5. An acetone solvate crystal form of 4- (4-guanidinobenzoyloxy) phenylacetic acid [2- (dimethylamino) -2-oxoethyl ] ester methanesulfonate shown as a formula (I),

characterized in that the X-ray powder diffraction expressed in 2 theta angles using Cu-K alpha radiation has diffraction peaks at 4.46, 8.89, 13.33, 15.33, 17.80 and 19.14 with an error of + -0.2.

6. The acetone solvate crystalline form according to claim 5 having an X-ray powder diffraction pattern with the following characteristic diffraction angles 2 θ, interplanar spacings d and relative intensities, wherein the error in 2 θ is 0.2:

preferably, the acetone solvate crystalline form has an X-ray powder diffraction pattern as shown in figure 9.

7. An N, N-dimethylformamide solvate crystal form of 4- (4-guanidinobenzoyloxy) phenylacetic acid [2- (dimethylamino) -2-oxoethyl ] ester methanesulfonate shown as a formula (I),

characterized in that the X-ray powder diffraction expressed in 2 theta angles using Cu-K alpha radiation has diffraction peaks at 14.92, 17.17, 18.17, 19.23, 20.04 and 20.33 with an error of + -0.2.

8. The crystalline form of N, N-dimethylformamide solvate according to claim 7, having an X-ray powder diffraction pattern with the following characteristic diffraction angles 2 θ, interplanar spacings d and relative intensities, wherein the error in 2 θ is 0.2:

preferably, the crystalline form of N, N-dimethylformamide solvate has an X-ray powder diffraction pattern as shown in figure 12.

9. A process for preparing a crystalline form of [2- (dimethylamino) -2-oxoethyl ] 4- (4-guanidinobenzoyloxy) phenylacetate mesylate according to claim 1 or 2, the process comprising:

(1) stirring free base 4- (4-guanidinobenzoyloxy) phenylacetic acid [2- (dimethylamino) -2-oxoethyl ] ester of the compound shown in the formula (I) in a reaction solvent, and dropwise adding a methanesulfonic acid aqueous solution;

(2) cooling, crystallizing and filtering to obtain crystals;

(3) and (3) heating and stirring the crystal obtained in the step (2) in water or an organic solvent, cooling and crystallizing, filtering the crystal, and drying to obtain the crystal.

10. The production method according to claim 9, wherein the reaction solvent in step (1) is one of water, aqueous methanol, aqueous ethanol, acetone, aqueous acetone, and N, N-dimethylformamide; preferably, the dropping temperature in the step (1) is 0-40 ℃, preferably 5-25 ℃, and more preferably 15-25 ℃;

preferably, the crystallization temperature in the step (2) is-10 to 5 ℃;

preferably, the organic solvent in step (3) is one of acetone, methanol, ethanol, isopropanol, tert-butanol, n-butanol, aqueous acetone and aqueous ethanol, preferably aqueous acetone or aqueous ethanol; preferably, the heating temperature in the step (3) is 40-80 ℃, preferably 50-60 ℃.

11. A method of preparing the hydrate crystalline form of claim 3 or 4, the acetone solvate crystalline form of claim 5 or 6, or the N, N-dimethylformamide solvate crystalline form of claim 7 or 8, the method of preparing comprising:

(1) stirring free base 4- (4-guanidinobenzoyloxy) phenylacetic acid [2- (dimethylamino) -2-oxoethyl ] ester of the compound shown in the formula (I) in a reaction solvent, and dropwise adding a methanesulfonic acid aqueous solution;

(2) cooling, crystallizing, filtering and drying.

12. The preparation method according to claim 11, wherein the dropping temperature in the step (1) is 0 to 40 ℃, preferably 5 to 25 ℃, and more preferably 15 to 25 ℃;

preferably, the crystallization temperature in the step (2) is-10 to 5 ℃;

wherein, when preparing the hydrate crystalline form of claim 3 or 4, the reaction solvent of step (1) is water, aqueous methanol or aqueous ethanol;

when preparing the acetone solvate crystalline form of claim 5 or 6, the reaction solvent of step (1) is acetone or aqueous acetone;

when preparing the crystalline form of N, N-dimethylformamide solvate according to claim 7 or 8, the reaction solvent in step (1) is N, N-dimethylformamide.

13. A pharmaceutical composition comprising the crystalline form of [2- (dimethylamino) -2-oxoethyl ] 4- (4-guanidinobenzoyloxy) phenylacetate mesylate of claim 1 or 2 and at least one pharmaceutically acceptable excipient; preferably, the pharmaceutical composition is an oral tablet or capsule; or

The pharmaceutical composition comprises the hydrate crystalline form of [2- (dimethylamino) -2-oxoethyl ] 4- (4-guanidinobenzoyloxy) phenylacetate methanesulfonate of claim 3 or 4 and at least one pharmaceutically acceptable excipient and/or diluent; preferably, the pharmaceutical composition is an oral tablet, capsule or injection.

14. Use of the crystalline form of [2- (dimethylamino) -2-oxoethyl ] 4- (4-guanidinobenzoyloxy) phenylacetate methanesulfonate of claim 1 or 2 or the crystalline form of hydrate of [2- (dimethylamino) -2-oxoethyl ] 4- (4-guanidinobenzoyloxy) phenylacetate methanesulfonate of claim 3 or 4 for the manufacture of a medicament for the treatment of pancreatitis, reflux esophagitis, and diseases caused by coronavirus infection, including middle east respiratory syndrome, severe acute respiratory syndrome, and novel coronavirus pneumonia.

Technical Field

The invention relates to crystal forms of camostat mesylate and solvates thereof, preparation methods thereof, pharmaceutical preparations thereof and uses thereof in the treatment of acute pancreatitis.

Background

Chemical name of Camostat mesylate (Camostat Mesilate): 4- (4-guanidinobenzoyloxy) phenylacetic acid [2- (dimethylamino) -2-oxoethyl ] ester methanesulfonate, having the formula:

the molecular formula is as follows: c₂₀H₂₂N₄O₅·CH₄O₃S, molecular weight: 494.52, CAS registry number: 57921-29-8.

Camostat mesylate is a non-peptide protease inhibitor for oral administration, can effectively inhibit the activity of trypsin, plasma kallikrein, plasmin, thrombin, C1r, C1 esterase and the like, and the compound has no obvious inhibition effect on alpha-chymotrypsin, pepsin, pancreatin and the like, so the camostat mesylate can be clinically applied to acute symptom relief of chronic pancreatitis, improvement of amylase values in blood and urine and treatment of alkaline reflux esophagitis caused by reflux of pancreatic juice and bile after gastrectomy.

The literature (cell.2020Apr 16; 181(2):271-280.) reports that a pharmaceutical composition comprising [2- (dimethylamino) -2-oxoethyl ] phenylacetate methanesulfonate represented by the formula (I) has a certain therapeutic effect on novel coronavirus pneumonia (COVID-19). There are also literature reports (Virology,2020,543,43-53, etc.) that they have potential therapeutic effects in other diseases caused by coronavirus infection, such as Middle East Respiratory Syndrome (MERS), Severe Acute Respiratory Syndrome (SARS), etc.

The camostat mesylate tablet is a synthetic protease inhibitor developed by the Japanese Ministry of pharmaceutical industry, is approved to be sold in Japan in 1986 by the Japan Ministry of health and labor, and is a white film coated tablet with the trade name of FOIPAN and the specification of 100 mg.

The preparation method of the camostat mesylate product has been reported at home and abroad, such as DE 2548886; JP 1976-5062; JP 1977-116655; JP 1996-149786; JP 2002114755; fine chemistry, 2004, (08), 597-; pharmaceutical and clinical studies, 2016,24(04), 321-; chemical development, 2010, 29(07),1334-1337, and the like.

The general synthetic method is as follows:

there are two melting points reported in the literature for this compound: 155 ℃ in 150-.

The above patent documents such as JP1976-5062, JP1977-116655, US4021472 and the like describe a preparation method of camostat mesylate, pharmaceutical compositions containing them and use for preparing a medicament for treating pancreatitis, but none of them relates to a crystal form of camostat mesylate. The preparation produced by Nippon Xiaoye corporation was subjected to thermal analysis, and the spectrum is shown in FIG. 1. It can be seen from the DSC spectrum that it has two endothermic peaks at 150 ℃ and 190 ℃. This is in contrast to the melting point of 150-155 ℃ reported in DE2548886 of Xiaoye and JP 1977-116655: 195 ℃ and 196 ℃. Therefore, the camostat mesylate prepared according to the prior literature is a mixed crystal form, which is not beneficial to the stability of the medicament.

The difference of water solubility of different crystal forms of the camostat mesylate is large, and the compound which has high purity, determined crystal form and good reproducibility is obtained in view of the pharmaceutical value of the compound, so that the important significance is achieved.

Disclosure of Invention

The inventors of the present invention have found one crystal form and several solvated crystal forms of camostat mesylate. By solvating the crystalline form, a stable single crystalline form is obtained. Compared with the mixed crystal form, the single crystal form and the hydrated crystal form have good stability, and the physicochemical properties of the crystal form are more stable in the preparation process than the mixed crystal form, so that the crystal form has more practical value. In addition, in the literature camostat mesylate preparation, seed crystals are usually added. Different crystal forms of the prepared camostat mesylate can be obtained without crystal seeds.

Through analysis comparison and experimental research of the literature, the camostat mesylate is mainly obtained by condensation reaction of an intermediate p-hydroxyphenylacetic acid-N, N-dimethylformamidoformyl methyl ester (3) and p-guanidinobenzoic acid hydrochloride (5). Intermediate 3 can be obtained from p-hydroxyphenylacetic acid (1) and 2-chloro-N, N-dimethylacetamide (2) or 2-bromo-N, N-dimethylacetamide. The synthesis method of the condensation reaction mainly comprises two methods: reacting 3 and 5 under the participation of a condensation reaction catalyst Dicyclohexylcarbodiimide (DCC) or 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI) to obtain camostat hydrochloride (6); and secondly, converting 5 into p-guanidinobenzoyl chloride hydrochloride (10) through an acylating agent such as thionyl chloride, oxalyl chloride (9) and the like, and condensing with the intermediate 3 to obtain camostat hydrochloride (6). The camostat hydrochloride (6) obtained in the above manner is further subjected to methanesulfonic acid in an intermediate state of the free base (11) to obtain the target product camostat mesylate (8, i.e., the compound of formula I). The camostat hydrochloride (6) can be precipitated into solid by adding sodium hydroxide solution, and is filtered and washed by water to obtain camostat free base (11).

The invention refers to the literature of medicine and clinical research, 2016,24(04),321-323 and the like, and adopts the following synthetic route:

the inventors of the present invention have conducted intensive studies and unexpectedly found that camostat mesylate has a polymorphism during salt formation and purification.

Therefore, it is an object of the present invention to provide camostat mesylate crystal forms A, C, E and F of the compound of formula (I) and a process for the preparation thereof and a process for interconversion between the different crystal forms.

Another object of the present invention is to provide camostat mesylate crystal form A, C of the compound of formula (I) as an active ingredient, and a pharmaceutical composition comprising one or more pharmaceutically acceptable carriers, excipients or diluents, and use thereof in the treatment of pancreatitis.

The present disclosure will now be described in detail for the purpose of the invention.

In one aspect, the invention provides a crystalline form (a) of [2- (dimethylamino) -2-oxoethyl ] 4- (4-guanidinobenzoyloxy) phenylacetate mesylate having the following chemical structure:

wherein the X-ray powder diffraction expressed by 2 theta angle using Cu-K alpha radiation has diffraction peaks with error of + -0.2 at 9.52, 12.16, 16.43, 17.10, 19.91, 20.40, 20.68 and 21.60.

According to the invention, there is provided crystalline form (a) having an X-ray powder diffraction pattern with the following characteristic diffraction angles 2 θ, interplanar spacings d and relative intensities (expressed as a percentage of the most intense radiation), wherein the error in 2 θ is 0.2:

according to the invention, a crystal form (A) is provided, which has the structure shown in figure 2¹H NMR spectrum.

The X-ray powder diffraction pattern of the crystal form (A) provided by the invention is shown in figure 3. The crystal form (A) provided by the invention has a DSC-TGA spectrum shown in figure 4.

The invention also provides a hydrate crystal form (C) of 4- (4-guanidinobenzoyloxy) phenylacetic acid [2- (dimethylamino) -2-oxoethyl ] ester methanesulfonate shown as a formula (I), wherein X-ray powder diffraction expressed by a 2 theta angle by using Cu-Kalpha radiation has diffraction peaks at 9.56, 12.14, 16.44, 17.10, 19.92, 20.39, 20.67 and 21.60, and errors of the diffraction peaks are +/-0.2.

According to the hydrate crystal form (C) provided by the invention, the X-ray powder diffraction pattern has the following characteristic diffraction angles 2 theta, interplanar distances d and relative intensities (expressed by the percentage of the strongest ray), wherein the error of the 2 theta is 0.2:

according to the invention, a hydrate crystal form (C) is provided, which is shown in figure 5¹H NMR spectrum. The X-ray powder diffraction pattern of the hydrate crystal form (C) provided by the invention is shown in figure 6. The hydrate crystal form (C) provided by the invention has a DSC-TGA spectrum shown in figure 7.

The invention also provides an acetone solvate crystalline form (E) of 4- (4-guanidinobenzoyloxy) phenylacetic acid [2- (dimethylamino) -2-oxoethyl ] ester methanesulfonate shown as formula (I), wherein X-ray powder diffraction expressed by 2 theta angle using Cu-Ka radiation has diffraction peaks at 4.46, 8.89, 13.33, 15.33, 17.80 and 19.14, and the error of the diffraction peaks is +/-0.2.

The acetone solvate crystal form (E) provided by the invention has an X-ray powder diffraction pattern with the following characteristic diffraction angles 2 theta, interplanar distances d and relative intensities (expressed by the percentage of the strongest rays), wherein the error of the 2 theta is 0.2:

according to the invention, an acetone solvate crystal form (E) is provided, which is shown as figure 8¹H NMR spectrum. The X-ray powder diffraction pattern of the acetone solvate crystal form (E) provided by the invention is shown in figure 9. The acetone solvate crystal form (E) provided by the invention has a DSC-TGA spectrum shown in figure 10.

The invention also provides a crystalline form of N, N-dimethylformamide solvate (F) of 4- (4-guanidinobenzoyloxy) phenylacetic acid [2- (dimethylamino) -2-oxoethyl ] ester methanesulfonate, wherein X-ray powder diffraction expressed in 2 theta angles using Cu-Ka radiation has diffraction peaks at 14.92, 17.17, 18.17, 19.23, 20.04, and 20.33, said diffraction peaks having an error of ± 0.2.

According to the invention, the N, N-dimethylformamide solvate form (F) is provided, the X-ray powder diffraction pattern of which has the following characteristic diffraction angles 2 theta, interplanar distances d and relative intensities (expressed as percentage of the strongest rays), wherein the error of 2 theta is 0.2:

according to the invention, the N, N-dimethylformamide solvate crystal form (F) is provided, and the N, N-dimethylformamide solvate crystal form (F) has the structure shown in figure 11¹H NMR spectrum. The X-ray powder diffraction pattern of the N, N-dimethylformamide solvate crystal form (F) provided by the invention is shown in figure 12. The N, N-dimethylformamide solvate crystal form (F) provided by the invention has a DSC-TGA spectrum shown in figure 13.

In another aspect, the invention also provides a preparation method of the camostat mesylate crystal forms A, C, E and F of the compound shown in the formula (I) and a method for interconversion between different crystal forms.

The invention also provides a preparation method of the crystal form (A) of the 4- (4-guanidinobenzoyloxy) phenylacetic acid [2- (dimethylamino) -2-oxoethyl ] ester mesylate, which comprises the following steps:

(1) stirring free base 4- (4-guanidinobenzoyloxy) phenylacetic acid [2- (dimethylamino) -2-oxoethyl ] ester of the compound shown in the formula (I) in a reaction solvent, and dropwise adding a methanesulfonic acid aqueous solution;

(2) cooling, crystallizing, filtering and drying to obtain crystals;

(3) and (3) heating and stirring the crystal obtained in the step (2) in water or an organic solvent, cooling and crystallizing, filtering the crystal and drying to obtain the crystal.

According to the preparation method of the crystal form (A), provided by the invention, the reaction solvent in the step (1) can be water or a water-containing hydrophilic organic solvent, and is preferably one of water, water-containing methanol (methanol concentration is 0-50 wt%), water-containing ethanol (ethanol concentration is 0-50 wt%), acetone, water-containing acetone (acetone concentration is 90-100 wt%) and N, N-dimethylformamide. Wherein the dropping temperature in the step (1) is 0-40 ℃, preferably 5-25 ℃, and more preferably 15-25 ℃.

According to the preparation method of the crystal form (A), provided by the invention, the crystallization temperature in the step (2) is-10-5 ℃.

The preparation method of the crystal form (A) provided by the invention is characterized in that the organic solvent in the step (3) is a hydrophilic organic solvent or a mixture of the hydrophilic organic solvent and water, preferably, the organic solvent is selected from one of acetone, methanol, ethanol, isopropanol, tert-butanol, n-butanol, hydrous acetone (acetone concentration is 90-100 wt%) and hydrous ethanol (ethanol concentration is 90-100 wt%), and more preferably, the hydrous acetone (acetone concentration is 90-100 wt%) or the hydrous ethanol (ethanol concentration is 90-100 wt%).

According to the preparation method of the crystal form (A), the heating temperature in the step (3) is 40-80 ℃, and preferably 50-60 ℃.

The invention also provides a preparation method of the hydrate crystal form (C), the acetone solvate crystal form (E) or the N, N-dimethylformamide solvate crystal form (F) of the 4- (4-guanidinobenzoyloxy) phenylacetic acid [2- (dimethylamino) -2-oxoethyl ] ester mesylate, wherein the preparation method comprises the following steps:

(1) stirring free base 4- (4-guanidinobenzoyloxy) phenylacetic acid [2- (dimethylamino) -2-oxoethyl ] ester of the compound shown in the formula (I) in a reaction solvent, and dropwise adding a methanesulfonic acid aqueous solution;

(2) cooling, crystallizing, filtering and drying.

The preparation method comprises the following steps of (1), wherein the dropping temperature in the step (1) is 0-40 ℃, preferably 5-25 ℃, and more preferably 15-25 ℃;

preferably, the crystallization temperature in the step (2) is-10 to 5 ℃.

Wherein, when the hydrate crystal form is prepared, the reaction solvent in the step (1) is water, water-containing methanol or water-containing ethanol;

when preparing the acetone solvate crystal form, the reaction solvent in the step (1) is acetone or aqueous acetone;

when the N, N-dimethylformamide solvate crystal form is prepared, the reaction solvent in the step (1) is N, N-dimethylformamide.

That is, form a of the present invention can be prepared by starting from the corresponding solvate form C, E, F.

In still another aspect, the present invention also provides a pharmaceutical composition comprising the crystalline form (a) and the hydrate crystalline form (C) as active ingredients and one or more pharmaceutically acceptable carriers, excipients or diluents, and use thereof in the preparation of a medicament for treating pancreatitis, reflux esophagitis, and diseases caused by coronavirus infection.

The invention provides a pharmaceutical composition, which comprises a crystal form (A) of 4- (4-guanidinobenzoyloxy) phenylacetic acid [2- (dimethylamino) -2-oxoethyl ] ester mesylate shown in a formula (I) and at least one medicinal excipient; preferably, the pharmaceutical composition is an oral tablet and/or capsule;

or the pharmaceutical composition comprises a hydrate crystal form C of 4- (4-guanidinobenzoyloxy) phenylacetic acid [2- (dimethylamino) -2-oxoethyl ] ester methanesulfonate shown as a formula (I) and at least one pharmaceutically-acceptable excipient and/or diluent and the like; preferably, the pharmaceutical composition is an oral tablet, capsule or injection.

The crystal form and the solvate crystal form provided by the invention can be used for preparing a medicament for treating acute pancreatitis and reflux esophagitis, and in addition, a document reports that a medicinal composition containing 4- (4-guanidinobenzoyloxy) phenylacetic acid [2- (dimethylamino) -2-oxoethyl ] ester methanesulfonate shown in a formula (I) has a certain treatment effect on novel coronavirus-induced pneumonia (COVID-19).

The invention also provides application of the crystal form (A) of the 4- (4-guanidinobenzoyloxy) phenylacetic acid [2- (dimethylamino) -2-oxoethyl ] ester mesylate or the hydrate crystal form (C) in preparing a medicament for treating pancreatitis, reflux esophagitis and diseases caused by coronavirus infection.

Wherein the diseases caused by coronavirus infection include middle east respiratory syndrome, severe acute respiratory syndrome and novel coronavirus pneumonia (COVID-19).

Drawings

Embodiments of the invention are described in detail below with reference to the attached drawing figures, wherein:

FIG. 1 is a thermogram spectrum of camostat mesylate product manufactured by Nippon Povid, batch 989 TC;

FIG. 2 shows the crystal form of camostat mesylate provided by the invention¹H NMR spectrum;

FIG. 3 is an X-ray powder diffraction pattern of a crystal form of camostat mesylate provided by the invention;

FIG. 4 is a DSC-TGA profile of a crystal form of camostat mesylate provided by the invention;

FIG. 5 shows a hydrate crystal form of camostat mesylate provided by the invention¹H NMR spectrum;

FIG. 6 is an X-ray powder diffraction pattern of a hydrate crystal form of camostat mesylate provided by the invention;

FIG. 7 is a DSC-TGA profile of a hydrate crystal form of camostat mesylate provided by the invention;

FIG. 8 shows acetone solvate crystal forms of camostat mesylate provided by the invention¹H NMR spectrum;

FIG. 9 is an X-ray powder diffraction pattern of the acetone solvate crystalline form of camostat mesylate provided by the invention;

FIG. 10 is a DSC-TGA profile of an acetone solvate crystalline form of camostat mesylate provided by the invention;

FIG. 11 shows a crystal form of N, N-dimethylformamide solvate of camostat mesylate provided by the invention¹H NMR spectrum;

FIG. 12 is an X-ray powder diffraction pattern of the N, N-dimethylformamide solvate crystalline form of camostat mesylate provided by the present invention;

fig. 13 is a DSC-TGA profile of the N, N-dimethylformamide solvate crystalline form of camostat mesylate provided by the invention.

Detailed Description

The present invention is described in further detail below with reference to specific embodiments, which are given for the purpose of illustration only and are not intended to limit the scope of the invention.

The hydrochloride of the 4- (4-guanidinobenzoyloxy) phenylacetic acid [2- (dimethylamino) -2-oxoethyl ] ester of formula (I) was prepared by the method of reference (pharmaceutical and clinical research, 2016,24(04), 321-323). Preparing 0.5mol/L solution by using 1.05 times of molar weight of sodium hydroxide, dripping the solution into the aqueous solution of the hydrochloride to obtain white solid, filtering, washing a filter cake by using water until dripping liquid drops are neutral, and obtaining the free base (hereinafter referred to as free base) of the 4- (4-guanidinobenzoyloxy) phenylacetic acid [2- (dimethylamino) -2-oxoethyl ] ester shown in the formula (I). The free base was allowed to air at room temperature overnight to give a dry free base, which was used as the starting material in the following examples. The stability of the free base is relatively poor and it is not suitable for long-term storage.

Example 1 preparation of hydrate form C

Adding 5g of the free alkali into 25ml of deionized water, cooling to 10-15 ℃, dropwise adding a methanesulfonic acid solution prepared from 1.2g of methanesulfonic acid and 1.5g of water, and keeping the temperature not to exceed 20 ℃ in the dropwise adding process. And after dripping, dissolving the system clearly, cooling to 0-5 ℃, stirring for 2h, and separating out solids. Filtering, leaching the filter cake with cold water, and drying for 6h by blowing at 50 ℃ to obtain 3.1g of the hydrate crystal form C of the 4- (4-guanidinobenzoyloxy) phenylacetic acid [2- (dimethylamino) -2-oxoethyl ] ester methanesulfonate of the compound of formula (I).

Example 2 preparation of acetone solvate form E

5g of the free base was added to a mixture of 45ml of acetone and 5ml of deionized water, and a suspension was obtained with stirring. Dripping methanesulfonic acid solution prepared by 1.2g of methanesulfonic acid and 2.4g of water at 15-25 ℃, and keeping the temperature not to exceed 25 ℃ in the dripping process. After dropping, the system was clear. And cooling to-10-0 ℃, and separating out solids. Stirring for 2h under heat preservation, filtering, leaching a filter cake with acetone, and drying for 4h by blowing at 50 ℃ to obtain 3.8g of the acetonide crystal form E of the compound 4- (4-guanidinobenzoyloxy) phenylacetic acid [2- (dimethylamino) -2-oxoethyl ] ester methanesulfonate of the formula (I).

Example 3 preparation of N, N-dimethylformamide solvate form F

5g of the free base was added to 15ml of N, N-dimethylformamide, and a suspension was obtained with stirring. Dripping 1.2g of methanesulfonic acid at 15-25 ℃, and keeping the temperature not to exceed 25 ℃ in the dripping process. After dropping, the system was clear. And cooling to 0-5 ℃, and separating out solids. Stirring for 2h under heat preservation, filtering, leaching a filter cake by using frozen N, N-dimethylformamide, and drying by blowing air at 90 ℃ for 5h to obtain 3.3g of N, N-dimethylformamide solvate crystal form F of 4- (4-guanidinobenzoyloxy) phenylacetic acid [2- (dimethylamino) -2-oxoethyl ] ester methanesulfonate of the compound of the formula (I).

Example 4 preparation of form A with hydrate form C

5g of the hydrate crystal form C prepared in example 1 is added into 25ml of 95% ethanol, and the mixture is heated to 50-60 ℃ under stirring, so that the hydrate crystal form C is gradually dissolved. And stopping heating after the system is dissolved and clear, and cooling and crystallizing. When the temperature is reduced to 30-40 ℃, solid is separated out. Further cooling to 0-5 ℃, keeping the temperature and crystallizing for 3 hours, and further increasing the precipitated solids. Filtering, and leaching a filter cake with 95% ethanol to obtain a white crystalline solid. Forced air drying at 60 ℃ for 6h gives 3.9g of 4- (4-guanidinobenzoyloxy) phenylacetic acid [2- (dimethylamino) -2-oxoethyl ] ester methanesulfonate of the formula (I) in the form of crystal A.

Example 5 preparation of form a from acetonide form E

5g of the acetone solvate form E prepared in example 2 was added to 50ml of 97% aqueous acetone (containing 48.5ml of acetone and 1.5ml of water), heated to 70-80 ℃ with stirring, and the form E was gradually dissolved. And stopping heating after the system is dissolved and clear, cooling to 0-5 ℃, and carrying out heat preservation and crystallization for 2 hours. Filtering, and leaching a filter cake with acetone to obtain a white crystalline solid. Air-blast drying at 80 ℃ for 6h gives 4.1g of 4- (4-guanidinobenzoyloxy) phenylacetic acid [2- (dimethylamino) -2-oxoethyl ] ester methanesulfonate of the formula (I) in crystalline form A.

Example 6 preparation of form a with N, N-dimethylformamide solvate form F

5g of the N, N-dimethylformamide solvate crystal form F prepared in example 3 is added into 15ml of deionized water, heated to 40-50 ℃ under stirring, and the crystal form F is gradually dissolved. And stopping heating after the system is dissolved and clear, cooling to 0-5 ℃, and carrying out heat preservation and crystallization for 2 hours. Filtering, and leaching a filter cake with cold water to obtain a white crystalline solid. Vacuum drying at 80 ℃ for 8h gave 3.5g of [2- (dimethylamino) -2-oxoethyl ] 4- (4-guanidinobenzoyloxy) phenylacetate methanesulfonate of the formula (I).

Example 7 preparation of a pharmaceutical composition of camostat mesylate form a

Preparing 1000 tablets in scale, adding 10mg of hydroxypropyl cellulose, 5mg of carboxymethyl cellulose calcium, 1mg of magnesium stearate, 5mg of polyoxyethylene, 3mg of polyoxypropylene, 3mg of ethylene glycol and 10mg of lactose hydrate into each tablet containing the camostat mesylate in the crystal form A, and directly tabletting and coating to prepare the camostat mesylate crystal form A tablet.

Example 8 preparation of a pharmaceutical composition of hydrate form C

Preparing 1000 capsules, each tablet containing 100mg of the C-crystal form of camostat mesylate, adding 10mg of hydroxypropyl cellulose, 5mg of carboxymethyl cellulose calcium, 1mg of magnesium stearate, 5mg of polyoxyethylene, 3mg of polyoxypropylene, 3mg of ethylene glycol and 10mg of lactose hydrate, and performing dry granulation to prepare the camostat mesylate hydrate crystal form C capsule.

Example 9 preparation of a pharmaceutical composition of hydrate form C

The preparation scale was 1000 bottles. The prescription composition is as follows:

camostat mesylate	10g
		Mannitol	10g
Citrate solution	Proper amount of
		Water for injection	2L

The preparation process comprises the following steps:

preparing a solution: dissolving the active ingredient camostat mesylate hydrate crystal form C in water for injection (at 25-30 ℃) to obtain a solution with the concentration of 10mg/ml, adding 10g of mannitol, stirring to dissolve, and adding a proper amount of citrate buffer solution to adjust the pH value to be within a range of 3.0-5.0. Adding injection water to a specified volume of 2L to ensure that the content of the camostat mesylate is 5mg/ml and the content of the mannitol is 5mg/ml, and freeze-drying to prepare the powder injection.

Comparative examples

The camostat mesylate crystal form a and camostat mesylate hydrate crystal form C tablets were prepared according to the method of example 7, and were placed at 70 ℃ under 93% RH for 1 month in the same manner as camostat mesylate tablets (lot: 989TC) produced by the japanese filoplume company, and the change of the main impurities and purity of the three were examined, and the results are shown in table 1.

TABLE 1

As can be seen from the data in table 1, compared with the mixed crystal form of camostat mesylate, the single crystal form a and the hydrate crystal form C have good stability, and the physicochemical properties of the crystal form a and the hydrate crystal form C are more stable in the preparation process than those of the existing crystal forms.