WO2018137502A1

WO2018137502A1 - Crystal form of salvianolic acid a, and method for preparing same

Info

Publication number: WO2018137502A1
Application number: PCT/CN2018/072432
Authority: WO
Inventors: 叶天健; 阚士东; 张志杰; 陆修伟; 贺元康; 曹焕英; 陈贵兰
Original assignee: 浙江永宁药业股份有限公司
Priority date: 2017-01-24
Filing date: 2018-01-12
Publication date: 2018-08-02
Also published as: CN108341747B; CN108341747A

Abstract

The present invention relates to the field of pharmaceutical chemistry, and in particular, to a crystal form of salvianolic acid A, and a method for preparing same. The X-ray powder diffraction pattern of a crystal form I shows characteristic peaks at 2θ values of 7.08 ± 0.2°, 11.94±0.2°, 14.23±0.2°, 16.41±0.2°, 17.99±0.2°, 21.41±0.2°, 21.83±0.2°, 22.47±0.2°, and 25.76±0.2°. The crystal form I of salvianolic acid A provided by the present invention has good stability, is not easy to be oxidized or deliquated under the influence of high temperature and high humidity, facilitates long-term storage of drugs, and can better avoid crystal transformation during drug research and development. Moreover, the purity of the crystal form I prepared in the present invention is higher than 99%, and is suitable as an active pharmaceutical ingredient (API) for research and development of pharmaceutical preparations.

Description

Crystal form of salvianolic acid A and preparation method thereof

Technical field

The invention relates to the field of medical chemistry, in particular to a crystal form of salvianolic acid A and a preparation method thereof.

Background technique

Salvianolic acid A is the most active water-soluble component of traditional Chinese medicine Salvia miltiorrhiza. It has been found to have a wide range of pharmacological activities, including anti-oxidation, myocardial ischemia protection, anti-thrombosis, neuroprotection, anti-fibrosis, prevention and treatment of diabetes, etc. In recent years, important advances have been made in the chemical studies, pharmacological activities, and pharmacokinetic studies of salvianolic acid A.

Salvianolic acid A is extremely unstable to light and heat, and it is easily oxidized to salvianolic acid C and isomalelic acid C when exposed to air, which is very unfavorable for long-term storage. Most of the prior art studies on the pharmacology and extraction process of salvianolic acid A, and there is almost no report on the solid crystalline substance of salvianolic acid A. In view of the wide medicinal value of salvianolic acid A, it is very necessary to seek a solid form of salvianolic acid A with good stability and high purity.

Summary of the invention

The invention relates to a crystal form I of salvianolic acid A, which has good stability, is not easily affected by high temperature and high humidity, and is oxidized or deliquescent, and is convenient for long-term storage. The invention also relates to preparation of salvianolic acid A crystal form I. method.

It is an object of the present invention to provide a new crystalline form of salvianolic acid A, designated as Form I. The X-ray powder diffraction pattern of the Form I has a 2θ value of 7.08±0.2°, 11.94±0.2°, 14.23±0.2°, 16.41±0.2°, 17.99±0.2°, 21.41±0.2°, 21.83±0.2°, There are characteristic peaks at 22.47±0.2° and 25.76±0.2°.

Further, the X-ray powder diffraction pattern of Form I provided by the present invention has a 2θ value of 12.78±0.2°, 13.61±0.2°, 13.93±0.2°, 20.07±0.2°, 24.25±0.2°, 24.57±0.2°, There are also characteristic peaks at 26.02±0.2°, 26.42±0.2°, 29.35±0.2°, and 36.11±0.2°.

Further, the X-ray powder diffraction data of the salvianolic acid A crystal form I of the present invention is as follows:

Another object of the present invention is to provide a preparation method of the crystalline form I of salvianolic acid A, which comprises: taking a sample of salvianolic acid A, completely dissolving it in a suitable solvent, and standing still at 0-25 ° C to obtain a crystal. Form I of salvianolic acid A.

The solvent is selected from the group consisting of water, methanol, ethanol, acetonitrile, acetone, acetic acid, ethyl acetate, butyl acetate, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), n-propanol, One of isopropyl alcohol; or the solvent is selected from the group consisting of water and a mixed solvent of one or more of methanol, ethanol, acetone, acetonitrile, acetic acid, DMF, DMSO.

Another object of the present invention is to provide another preparation method of the crystalline form I of salvianolic acid A, comprising the steps of: taking a sample of salvianolic acid A, completely dissolving it in a polar solvent to obtain a salvianolic acid A solution, and then Adding a low-polar solvent to the solution until crystals are precipitated, and statically crystallizing at 0-25 ° C to obtain crystal form I of salvianolic acid A;

The polar solvent is selected from the group consisting of methanol, ethanol, acetonitrile, acetone, acetic acid, ethyl acetate, butyl acetate, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), n-propanol, One or more of isopropanol, the low polar solvent being selected from one or more of toluene, tetrahydrofuran, n-hexane, cyclohexane, chloroform, petroleum ether.

Preferably, the polar solvent is selected from the group consisting of methanol, ethanol, acetonitrile, acetone, acetic acid, ethyl acetate, butyl acetate, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), and positive One of propanol and isopropanol, the low polar solvent being selected from the group consisting of toluene, tetrahydrofuran, n-hexane, cyclohexane, chloroform, and petroleum ether.

Further, the salvianolic acid A sample of the present invention is prepared by the method disclosed in the patent ZL 201310487751.6, and the main steps are as follows:

Step 1: Adding an extractant to the Salvia miltiorrhiza sinensis. After heating and extracting, the extract is concentrated by filtration to obtain a Salvia miltiorrhiza extract;

Step 2: Adjust the pH of Salvia miltiorrhiza extract to 4-8, concentrate under reduced pressure to 10%-30% of the original volume of Salvia miltiorrhiza extract to obtain concentrated Salvia miltiorrhiza extract. After adding hydrotalcite to concentrated Salvia miltiorrhiza extract, at 70-100 °C Heating the reaction for 5-40h to obtain a crude solution of salvianolic acid A;

Step 3: Adjust the pH of the crude solution of salvianolic acid A to 6-8. After adsorption by the reversed-phase resin, elute with water, then elute with aqueous methanol or ethanol solution, collect the eluate step by step, and combine to obtain dansyl alcohol. Sample solution of acid A.

Step 4: The salvianolic acid A sample solution is concentrated and dried to obtain salvianolic acid A powder.

The salvianolic acid A crystal form I provided by the invention has good stability, is not easily affected by high temperature and high humidity, undergoes oxidation or deliquescence, is convenient for long-term storage of the medicine, and can better avoid the phenomenon of crystal transformation during drug development; The purity of the crystal form I prepared by the invention is higher than 99%, and is suitable as a raw material medicine (API) for the development of pharmaceutical preparations.

DRAWINGS

Figure 1 is an XRPD pattern of salvianolic acid A crystal form I

Figure 2 shows the DSC curve of salvianolic acid A crystal form I

Figure 3 is an HPLC chromatogram of salvianolic acid A lyophilized powder (99.5%) and salvianolic acid A crystal form I (99.9%)

detailed description

The invention is further illustrated by the following examples, which are to be construed as illustrative and not restrictive.

The English abbreviations used in the present invention are explained as follows:

XRPD: X-ray powder diffraction

DSC: Differential Scanning Calorimetry

HPLC: high performance liquid chromatography

RH: relative humidity

Lx: lux (light intensity unit)

If abbreviations not included in the above list appear in the present invention, they are meant to be generally accepted by those skilled in the art.

Example 1

1 g of salvianolic acid A sample (purity 99.5%) was dissolved in 10 mL of water (water temperature 50 ° C), and the solution was cooled to 4 ° C overnight. The crystals were precipitated and allowed to stand for 72 h, and more crystals were observed. The crystals were filtered, and placed in a desiccator containing anhydrous calcium chloride as a drying agent for 24 hours at room temperature to obtain 375 mg of crystalline powder (Form I). The purity was determined by HPLC to be 99.9%. This crystal can be used for the core of other crystals. The X-ray powder diffraction data of Form I obtained in this example is shown in Table 1, the XRPD pattern thereof is shown in Fig. 1, and the DSC spectrum is shown in Fig. 2.

Table 1 X-ray powder diffraction data of salvianolic acid A crystal form I

Example 2

1 g of salvianolic acid A sample (purity 97.0%) was dissolved in 10 mL of water (water temperature 50 ° C), cooled to room temperature, and 10.2 mg of the pulverized salvianolic acid A nucleus (obtained by the method of Example 1) was added. After gently shaking, the temperature was lowered to 4 ° C and allowed to stand for 72 h, and crystals were observed to precipitate. The crystals were filtered, and placed in a desiccator containing anhydrous calcium chloride as a drying agent for 24 hours at room temperature to obtain 657 mg of crystalline powder (Form I). The purity was determined by HPLC to be 99.8%.

Example 3:

Take 1 mg of salvianolic acid A sample (purity 97.0%), dissolve it in 5 mL of ethanol, stir constantly, add 8 mL of toluene dropwise, and add 10.5 mg of the pulverized salvianolic acid A nucleus (obtained by the method of Example 1). After gently shaking, the temperature was lowered to 4 ° C and allowed to stand for 72 hours. Crystallization was observed, and the crystals were filtered, and placed in a desiccator containing anhydrous calcium chloride as a desiccant to dry under vacuum at 40 ° C for 24 hours to obtain 598 mg of a crystalline powder. The purity was determined by HPLC to be 99.8%.

Example 4:

1 g of salvianolic acid A sample (purity 97.0%) was dissolved in 5 mL of methanol, stirring was continued, 7 mL of toluene was added dropwise, and 10.0 mg of the pulverized salvianolic acid A core was added (the method of Example 1 was used). After gently shaking, the temperature was lowered to 4 ° C and allowed to stand for 72 hours. Crystallization was observed, and the crystals were filtered, and placed in a desiccator containing anhydrous calcium chloride as a desiccant to dry under vacuum at 40 ° C for 24 hours to obtain 556 mg of a crystalline powder. The purity was determined by HPLC to be 99.9%.

Example 5:

1 g of salvianolic acid A sample (purity 97.0%) was dissolved in 5 mL of acetone, stirring was continued, 10 mL of toluene was added dropwise, and 9.8 mg of the pulverized salvianolic acid A mother core was added (the method of Example 1 was used). After gently shaking, the temperature was lowered to 4 ° C and allowed to stand for 72 hours. Crystallization was observed, and the crystals were filtered, and placed in a desiccator containing anhydrous calcium chloride as a desiccant to dry under vacuum at 40 ° C for 24 hours to obtain 623 mg of a crystalline powder. The purity was determined by HPLC to be 99.9%.

Example 6

1 g of salvianolic acid A sample (purity 97.0%) was dissolved in 5 mL of ethanol, stirring was continued, 8 mL of tetrahydrofuran was added dropwise, and 11.3 mg of the pulverized salvianolic acid A nucleus was added (the method of Example 1 was obtained). After gently shaking, the temperature was lowered to 4 ° C and allowed to stand for 72 hours. Crystallization was observed, and the crystals were filtered, and placed in a desiccator containing anhydrous calcium chloride as a desiccant to dry under vacuum at 40 ° C for 24 hours to obtain 518 mg of a crystalline powder. The purity was determined by HPLC to be 99.9%.

Example 7

High temperature test of salvianolic acid A crystal form I

Take a sample of salvianolic acid A (lyophilized powder, amorphous, purity 99.5%) and salvianolic acid A crystal form I (purity 99.9%, obtained by the method of Example 1) 50mg each, tiled in a weighing bottle Medium, placed in a constant temperature oven, kept at 60 ° C for 10 days. After 10 days, the samples were compared with the same batch samples stored at -20 ° C, and the HPLC purity and peak area (converted to the unit mass peak area) were measured, and the purity changes and degradation of the two samples were monitored. The results are as follows:

It was determined that the crystal form of salvianolic acid A crystal form I was unchanged before and after the test.

Example 8

High-humidity test of salvianolic acid A crystal form I

Take a sample of salvianolic acid A (lyophilized powder, amorphous, purity 99.5%) and salvianolic acid A crystal form I (purity 99.9%, obtained by the method of Example 1) 50 mg each, and lay it in a weighing bottle The cells were placed in a closed container containing a saturated sodium chloride solution (relative humidity: 75% ± 1%) and allowed to stand at 25 ° C for 10 days. After 10 days, the samples were compared with the two samples before the test, and the HPLC purity and peak area (converted to unit mass peak area) and water content were measured, and the purity changes and moisture absorption of the two samples were monitored. The results are as follows:

The results showed that the sample of salvianolic acid A was more hygroscopic when it was placed under the condition of 25 °C and 75%±1% RH for a long time, and it showed oil droplets. However, the water content of the crystal form I provided by the present invention was only 1.89%, and its morphology was not obvious change.

Example 9

Strong light irradiation test of salvianolic acid A crystal form I

Take a sample of salvianolic acid A (lyophilized powder, amorphous, purity 99.5%) and salvianolic acid A crystal form I (purity 99.9%, obtained by the method of Example 1) 50mg each, tiled in a weighing bottle In the case, the illuminance was placed at 4,500 lx ± 500 lx for 10 days. After 10 days, the samples were compared with the same batch of light-shielded samples, and the HPLC purity and peak area (converted to unit mass peak area) were measured to monitor the purity and degradation of the two samples. The results are as follows:

Claims

A crystalline form I of salvianolic acid A, characterized in that the X-ray powder diffraction pattern of the crystalline form I has a 2θ value of 7.08±0.2°, 11.94±0.2°, 14.23±0.2°, 16.41±0.2°, There are characteristic peaks at 17.99±0.2°, 21.41±0.2°, 21.83±0.2°, 22.47±0.2°, and 25.76±0.2°.
The crystal form I according to claim 1, wherein the X-ray powder diffraction pattern has a 2θ value of 12.78±0.2°, 13.61±0.2°, 13.93±0.2°, 20.07±0.2°, 24.25±0.2°. Characteristic peaks at 24.57±0.2°, 26.02±0.2°, 26.42±0.2°, 29.35±0.2°, and 36.11±0.2°.
The crystal form I according to claim 1, wherein the X-ray powder diffraction data of the salvianolic acid A crystal form I is as follows:
The method for preparing a crystalline form I of salvianolic acid A according to any one of claims 1 to 3, characterized in that a sample of salvianolic acid A is taken and completely dissolved in a solvent to obtain a solution of salvianolic acid A, 0-25 ° C The crystal is allowed to stand to obtain the crystalline form I of salvianolic acid A; the solvent is selected from the group consisting of water, methanol, ethanol, acetonitrile, acetone, acetic acid, ethyl acetate, butyl acetate, N,N-dimethylformamide, and One of sulfoxide, n-propanol, isopropanol; or the solvent is selected from the group consisting of water and methanol, ethanol, acetone, acetonitrile, acetic acid, N,N-dimethylformamide, dimethyl sulfoxide A mixed solvent of one or several components.
The method for preparing a crystalline form I of salvianolic acid A according to claim 4, wherein a sample of salvianolic acid A is taken and completely dissolved in water at 50 ° C to obtain a solution of salvianolic acid A, and the temperature is lowered to 4 ° C. After 72 h, the dried Form I of Salvianolic Acid A was filtered.
The method for preparing a crystalline form I of salvianolic acid A according to any one of claims 1 to 3, characterized in that a sample of salvianolic acid A is taken and completely dissolved in a polar solvent to obtain a salvianolic acid A solution, and then Adding a low-polar solvent to the solution to precipitate crystals, and crystallization at 0-25 ° C to obtain crystal form I of salvianolic acid A; the polar solvent is selected from the group consisting of methanol, ethanol, acetonitrile, acetone, acetic acid, and acetic acid One or more of ester, butyl acetate, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), n-propanol, isopropanol, the low polar solvent selected from the group consisting of One or more of toluene, tetrahydrofuran, n-hexane, cyclohexane, chloroform, petroleum ether.
The preparation method according to any one of claims 4 to 6, characterized in that the salvianolic acid A solution is added to the salvianolic acid A mother nucleus and then crystallized.
The process according to claim 7, wherein the salvianolic acid A core is prepared according to the method of claim 4 or 5 or 6.