Danshensu sodium II crystal form and preparation method thereof

Abstract

The present invention discloses a sodium danshensu II crystal form and a preparation method thereof. The sodium danshensu II crystal form: #imgabs0# wherein the X-ray powder diffraction pattern of the sodium danshensu II crystal form is 2 θ =9.860, 14.000, 15.900, 16.900, 19.260, 23.480, 24.520, 25.220, 26.480, 28.180, 29.520, 29.960, 30.440, 31.880, 32.460, 37.100, 37.680 and 40.620, among which 2 θ The error range of the value is ±0.2. The tanshinone sodium II crystal form provided by the present invention has the advantages of high solubility, good stability, etc., is more conducive to preparation development, and is suitable for industrial production.

Description

Technical Field

[0001] The present invention relates to sodium danshensu polymorphs, in particular to sodium danshensu II crystal form and a preparation method thereof, as well as active drugs and pharmaceutical compositions related to the crystal form. Background Art

[0002] The chemical name of Danshensu is [D-(+)-β-(3,4-dihydroxyphenyl) lactic acid]. Due to its unstable structure, it is usually preserved as sodium salt. The molecular formula of Danshensu sodium is C9H9O5Na, the molecular weight is 220.15, and the structural formula is:

[0003]

[0004] Danshensu is one of the main active ingredients of Danshen, and domestic and foreign scholars have conducted a lot of research on its pharmacological effects. It is found that Danshensu has the effects of scavenging free radicals, anti-oxidation, anti-inflammation, anti-fibrosis, anti-lipid peroxidation, anti-apoptosis, anti-autophagy, and anti-Ca 2+ It has multiple pharmacological activities such as overload, platelet aggregation inhibition, thrombosis inhibition, atherosclerosis prevention, vascular endothelial cell protection, myocardial ischemia and hypoxia protection, coronary dilation, endothelial progenitor cell function improvement, microcirculation improvement, mitochondrial function improvement, immune regulation of fat metabolism, etc. It has protective effects on cardiovascular and cerebrovascular, liver, kidney, and lung. It is one of the main components of the traditional Chinese medicine Danshen for the treatment of cardiovascular and cerebrovascular diseases (Zhang Jinli, et al. Eur J Pharmacol. 2019, 864: 172710; Bao Xiaoyi, et al. Front Pharmacol. 2018, 9: 1445). The wide range of pharmacological effects of Danshensu suggests that Danshensu and its sodium salt have broad potential therapeutic prospects for the treatment of various cardiovascular and cerebrovascular diseases, and are worthy of clinical expansion and further development.

[0005] As we all know, polymorphism is common in solid drugs, and polymorphism is one of the important factors affecting the quality and efficacy of solid drugs. Different crystal forms of the same drug have different mechanical, thermodynamic, physical and chemical properties, and may have significant differences in appearance, solubility, melting point, dissolution, bioavailability, etc., which will affect its dissolution and absorption in the body, and further affect the bioavailability and efficacy of the drug, and even produce adverse reactions, affecting the safety of medication.

[0006] Therefore, the crystal form is of great significance for drug development. However, so far, there is no literature reporting the characteristics of the crystal form of Danshensu sodium and its preparation method, as well as drug combinations containing the crystal form. Summary of the invention

[0007] The purpose of the present invention is to provide a sodium danshensu II crystal form and a preparation method thereof. The sodium danshensu II crystal form provided by the present invention has the advantages of high solubility and good stability, is more conducive to formulation development, and is suitable for industrial production.

[0008] The present invention provides a sodium danshensu II crystal form, the X-ray powder diffraction pattern of the crystal form is 2 θ =9.860, 14.000, 15.900, 16.900, 19.260, 23.480, 24.520, 25.220, 26.480, 28.180, 29.520, 29.960, 30.440, 31.880, 32.460, 37.100, 37.680 and 40.620, among which 2 θ The value error range is ±0.2.

[0009] Furthermore, the X-ray powder diffraction pattern of the Danshensu sodium II crystal form is 2 θ There are diffraction peaks at =9.860, 10.900, 11.520, 14.000, 14.700, 15.900, 16.900, 17.780, 18.380, 19.260, 20.380, 20.940, 22.480, 22.980, 23.480, 24.520, 25.220, 26.480, 28.180, 29.520, 29.960, 30.440, 31.880, 32.460, 33.120, 35.800, 36.460, 37.100, 37.680, 38.860, 40.620 and 43.480.

[0010] Furthermore, the tanshinone sodium II crystal form has the same Figure 1 Essentially identical XRPD patterns.

[0011] The present invention also provides a method for preparing the above-mentioned Danshensu Sodium II crystal form, comprising:

[0012] 1) a. Take Danshensu sodium raw material, add water and organic solvent, heat and stir to dissolve;

[0013] Or b. Take the sodium danshensu raw material, add water or an organic solvent for the first time, heat and stir to dissolve; then add water or an organic solvent for the second time and continue to stir and dissolve; wherein, when water is added for the first time, an organic solvent is added for the second time; if an organic solvent is added for the first time, water is added for the second time;

[0014] 2) Keep stirring at room temperature until a large amount of crystals are precipitated, filter and dry to obtain sodium danshensu crystals.

[0015] Furthermore, the mass volume ratio of the sodium danshensu raw material to water in the step 1) is 1 g: (1-30) mL.

[0016] Furthermore, the organic solvent in step 1) can be an organic solvent or a combination of ketones, ethers, alkanes, aromatic hydrocarbons, esters, nitriles, alcohols or halogenated hydrocarbons, preferably one or more of acetone, methyl ethyl ketone, acetonitrile, ethyl acetate, ether, tert-butyl ether, petroleum ether, tetrahydrofuran, dichloromethane, methanol, ethanol, isopropanol and n-butanol; wherein the amount of the organic solvent used is 0.5 to 5 times the amount of water used.

[0017] Furthermore, the temperature of the heating, stirring and dissolving in step 1) is controlled at 10 to 70°C, preferably 20 to 45°C.

[0018] Furthermore, the temperature for adding water or organic solvent for the second time in step 1) b and continuing to stir and dissolve is controlled at 0 to 50° C., preferably 20 to 40° C.

[0019] Furthermore, the stirring time at room temperature is controlled to be 12 to 48 hours, preferably 24 hours.

[0020] Furthermore, the drying treatment is specifically drying under reduced pressure in a vacuum oven at 80° C. for 24 hours or drying under reduced pressure using an oil pump for 12 hours.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] Experimental studies have shown that the sodium danshensu II crystal form prepared by the present invention has better solubility in aqueous solution and faster dissolution rate in PBS solution, which is beneficial to dissolution and absorption in the human body. Its good solubility characteristics make the new crystal form have great application prospects.

[0023] The sodium danshensu II crystal form prepared by the present invention has been found in experimental studies that the new crystal form is stored under high humidity (25°C / 90% humidity) conditions, high temperature (60°C / air humidity) and light (light intensity 4500lux±500lux) conditions. The new sodium danshensu crystal form obtained by the present invention has no obvious changes in content, appearance and crystal form, has good stability, and can be better applied to pharmaceutical preparations. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 It is an X-ray powder diffraction spectrum (XRPD) of the Danshensu Sodium II crystal form prepared by the present invention;

[0025] Figure 2 This is a thermogravimetric analysis (TG) diagram of the Danshensu Sodium II crystal form prepared by the present invention;

[0026] Figure 3It is a differential scanning calorimetry (DSC) diagram of the Danshensu Sodium II crystal form prepared by the present invention;

[0027] Figure 4 It is an infrared spectrum (IR) diagram of the Danshensu Sodium II crystal form prepared by the present invention;

[0028] Figure 5 This is a hydrogen nuclear magnetic spectrum (H NMR) of the Danshensu sodium II crystal form prepared by the present invention;

[0029] Figure 6 It is a high performance liquid chromatography (HPLC) diagram of the Danshensu Sodium II crystal form prepared by the present invention;

[0030] Figure 7 The stability X-ray powder diffraction (XRPD) diagram of the sodium danshensu II crystal prepared by the present invention at 0d and 10d under high humidity conditions;

[0031] Figure 8 The stability X-ray powder diffraction (XRPD) diagram of the sodium danshensu II crystal prepared by the present invention at 0d and 10d under high temperature conditions;

[0032] Fig. 9 The present invention is an X-ray powder diffraction (XRPD) diagram of the stability of the sodium danshensu II crystal form prepared by the present invention at 0d and 10d under illumination conditions. DETAILED DESCRIPTION

[0033] The present invention is described in detail below in conjunction with specific embodiments. The following embodiments will help those skilled in the art to further understand the present invention, but the following embodiments should not be regarded as limiting the present invention. It should be noted that, for those of ordinary skill in the art, some adjustments and improvements can also be made without departing from the concept of the present invention. These all belong to the protection scope of the present invention.

[0034] The sodium danshensu used in the following examples is prepared according to the Chinese patent application number 202110184726.5, entitled “A process for extracting high-purity sodium danshensu”, the entire contents of which are incorporated herein by reference.

[0035] X-ray powder diffraction (XRD) analysis: Japanese Rigaku D / Max-2500 model: radiation source: copper target scanning at room temperature: scanning range 2.0-50.0°, step length: 0.02°, scanning rate: 8° / min;

[0036] Thermogravimetric analyzer (TG) analysis: Swiss Mettler Toledo TGA2 thermogravimetric analyzer, scanning range 30 ~ 180 ° C test weight loss curve, heating rate set to 10 ° C min -1 , nitrogen protection;

[0037] Differential scanning calorimetry (DSC) analysis: Swiss Mettler-Toledo DSC1, 30-280°C range, heating rate: 5°C / min, nitrogen protection;

[0038] Infrared spectroscopy (IR) analysis: The samples were scanned by Spectrum 400 from PerkinElmer, USA, with a spectral scanning range of 4000-500cm -1 , resolution 4.000cm -1 , scanning times 16 times, measured by attenuated total reflection technique (ATR);

[0039] High performance liquid chromatography (HPLC) purity detection conditions: octadecylsilane bonded silica gel as filler (150 mm × 4.6 mm, 3 μm); mobile phase 0.5% glacial acetic acid solution-methanol (90:10); column temperature 28°C; detection wavelength 280 nm;

[0040] H NMR analysis: Bruker 600 MHz NMR spectrometer, deuterated water as solvent.

[0041] The experimental methods in the following examples are all conventional methods unless otherwise specified.

[0042] Example 1: Preparation of Danshensu Sodium II Crystalline Form

[0043] Take 10g of sodium danshensu raw material, add 20mL of purified water, heat to 50℃ to dissolve, add 40mL of ethanol, stir at this temperature for 1 hour, then slowly cool to room temperature, stir and crystallize for 12h, a large amount of crystals precipitate, filter; the filter cake is dried under reduced pressure in a vacuum oven at 70℃ for 24 hours to obtain 8.7g of sodium danshensu crystals, with a yield of 87% and an HPLC purity of more than 99.85%.

[0044] The sample prepared in this example was sampled and XRD was performed. Figure 1 As shown, the obtained crystal form is crystal form II. The specific peak positions are shown in Table 1 below.

[0045] Table 1: X-ray powder diffraction data of Danshensu Sodium II crystal form in Example 2 of the present invention

[0046]

[0047]

[0048] Those skilled in the art should understand that in this embodiment, the diffraction peaks in the table do not represent the detailed situation of the diffraction peaks shown by the Danshensu Sodium II crystal form. The 2θ value of the X-ray powder diffraction pattern can vary slightly with the machine and with changes in sample preparation and batch-to-batch changes, and the quoted values ​​are not considered absolute values. It should also be understood that the relative intensity of the peaks may vary with orientation effects, so the intensity shown in the XRPD traces contained in the present invention is exemplary and is not used for absolute comparison.

[0049] The crystal samples obtained in this example were subjected to thermogravimetric analysis, differential scanning calorimetry and infrared spectroscopy tests. The obtained TG spectra, DSC thermal spectra and infrared spectra were as follows: Figure 2 , Figure 3 and Figure 4 shown.

[0050] The preparation method of this embodiment first uses water to dissolve sodium danshensu, and then adds an organic solvent to crystallize sodium danshensu. By using water and an organic solvent together, sodium danshensu with a new crystal form is successfully prepared, and the obtained sodium danshensu crystals are of high purity. The purity can reach 99.85% by HPLC detection, and the maximum single impurity content is extremely low, less than 0.01%. HPLC liquid phase purity is as follows Figure 5 shown.

[0051] Product 1 H-NMR spectrum Figure 6 As shown, the NMR data are as follows:

[0052] 1 H-NMR (600MHz, D2O) δ6.90 (d, J=8.4Hz, 1H), 6.84 (d, J=1.8Hz, 1H), 6.78 (dd, J=8.4,1.8 Hz, 1H), 4.24-4.22 (m, 1H), 3.02 (dd, J=14.4, 4.2Hz, 1H), 2.81 (dd, J=14.4, 7.8Hz, 1H).

[0053] Example 2: Preparation of Danshensu Sodium II Crystalline Form

[0054] Take 10g of sodium danshensu raw material, add 30mL of ethanol, 5mL of n-butanol, and then add 40mL of purified water, heat to 60℃ to dissolve, keep stirring for 1 hour, then slowly cool to 12℃, stir and crystallize for 12h, a large amount of crystals precipitate, filter; place the filter cake in 20mL of ethanol, stir for 8h, and filter; the filter cake is vacuum dried at 60℃ to obtain sodium danshensu crystals, and the results of X-ray powder diffraction (XRPD) analysis show that the obtained crystal form is crystal form II.

[0055] Example 3: Study on the high humidity stability of Danshensu sodium II crystal form

[0056] The tanshinone sodium II crystal solid prepared in Example 1 was placed under high humidity (25°C / 90% humidity) for 10 days, and samples were taken on the 0th and 10th days. HPLC analysis was performed to determine the purity change of the experimental sample; XRPD was used for X-ray powder diffraction detection to determine the crystal form change of the experimental sample (such as Figure 7 The results showed that the Danshensu sodium II crystal form was stable under high humidity conditions (as shown in Table 2).

[0057] Example 4: Investigation of high temperature stability of Danshensu sodium II crystal form

[0058] The tanshinone sodium II crystal solid prepared in Example 1 was placed under high temperature (60°C / air humidity) for 10 days, and samples were taken on the 0th and 10th days. HPLC analysis was performed to determine the purity change of the experimental sample; XRPD was used for X-ray powder diffraction detection to determine the crystal form change of the experimental sample (such as Figure 8 See Table 2 for details on the stability results of sodium danshensu II crystal form. The results show that sodium danshensu II crystal form is stable under high temperature conditions (as shown in Table 2).

[0059] Example 5: Study on the Light Stability of Danshensu Sodium II Crystalline Form

[0060] The solid form of Danshensu sodium II prepared in Example 1 was placed under a light intensity of 4500 lux ± 500 lux for 10 days, and samples were taken on the 0th and 10th days. HPLC analysis was performed to determine the purity change of the experimental sample; XRPD was used for X-ray powder diffraction detection to determine the crystal form change of the experimental sample (such as Fig. 9 The results showed that the crystalline form of Danshensu Sodium II was stable under light conditions (as shown in Table 2).

[0061] Table 2: Stability of Danshensu Sodium II Crystalline Form

[0062]

[0063]

[0064] The experimental results of Examples 3-5 show that:

[0065] Under the experimental conditions in the above table, the sodium danshensu II crystal form obtained by the present invention has no obvious changes in content, appearance and crystal form, has good stability, and can be better applied to pharmaceutical preparations.

[0066] Application examples: liquid dosage forms (such as injections, solutions, etc.), solid dosage forms (such as tablets, capsules, etc.), semi-solid dosage forms (such as ointments, gels, etc.), gas dosage forms (such as aerosols, sprays, etc.) of Danshensu Sodium II crystal form

[0067] Injection: Add 20 g of Danshensu Sodium II crystal form into 1 L of water for injection to dissolve, add 10 g of sodium chloride as an osmotic pressure regulator, adjust the pH value to 7.0-8.0 with dilute hydrochloric acid, filter through a 0.22 μm filter membrane, divide into packages, and sterilize at 121°C for 30 minutes.

[0068] Tablets: Add 10 grams of Danshensu Sodium II crystal form into 400 grams of lactose and mix thoroughly, add 30% PVP binder for granulation, dry, and then press into tablets.

[0069] Capsules: Add 10 g of Danshensu Sodium II crystal form to 100 g of microcrystalline cellulose and 300 g of starch and mix thoroughly, add 30% PVP binder for granulation, dry, and fill capsules to obtain the product.

[0070] Ointment: Heat 300g of vaseline, 100g of higher fatty alcohol, and 50g of glyceryl monostearate to 80℃ to melt, and filter with fine cloth; dissolve 5g of sodium lauryl sulfate in water, heat to 95℃, and slowly add the aqueous solution to the oil phase while stirring to make an emulsion base. Add 10g of sodium danshensu II crystal form and stir until condensed, and divide into packages to obtain the ointment.

[0071] Spray: Dissolve 9g of sodium chloride in 800mL of water for injection, add 20g of Danshensu Sodium II crystal to dissolve, adjust the pH to 7.0-8.0 with dilute hydrochloric acid, and add water for injection to 1000mL. Filter the prepared solution with a 0.22μm sterilizing filter, then dispense into the nasal spray device bottle and cover it.

[0072] Comparative Example: Solubility Comparison of Danshensu Sodium II Crystalline Form and Commercially Available Danshensu Sodium

[0073] Sources of the tested samples: Danshensu sodium II crystal form prepared in Example 1, with a purity greater than 99%; Danshensu sodium available on the market with a purity greater than 98%.

[0074] Experimental method: Sodium Danshensu II crystal form and commercially available sodium Danshensu were fully dissolved in pH 6.8 phosphate buffered saline (see the Chinese Pharmacopoeia for preparation method), and the concentration of each solution was detected by HPLC until its concentration no longer increased. The final concentration was recorded and the solubility of each crystal form was calculated accordingly.

[0075] Experimental results:

[0076] Table 3: Solubility of different crystalline forms of Danshensu sodium

[0077] Crystal form Solubility (pH 6.8 phosphate buffered saline) Danshensu Sodium Ⅱ Crystal Form 218.76mg / mL Danshensu Sodium 157.30mg / mL

[0078] According to the above results, the solubility of Danshensu sodium II crystal form is better than that of commercially available Danshensu sodium.

[0079] The preferred embodiments of the present invention have been specifically described above, but the present invention is not limited to the embodiments. Those skilled in the art may make various equivalent modifications or substitutions without violating the spirit of the present invention. These equivalent modifications or substitutions are all included in the scope defined by the claims of this application.

Claims

1. A tanshinone sodium II crystal form, characterized in that: The X-ray powder diffraction pattern of the Danshensu sodium II crystal form is 2 θ =9.860, 14.000, 15.900, 16.900, 19.260, 23.480, 24.520, 25.220, 26.480, 28.180, 29.520, 29.960, 30.440, 31.880, 32.460, 37.100, 37.680 and 40.620, among which 2 θ The value error range is ±0.

2.

2. The tanshinone sodium II crystal form according to claim 1, characterized in that: The X-ray powder diffraction pattern of the Danshensu sodium II crystal form is 2 θ There are diffraction peaks at =9.860, 10.900, 11.520, 14.000, 14.700, 15.900, 16.900, 17.780, 18.380, 19.260, 20.380, 20.940, 22.480, 22.980, 23.480, 24.520, 25.220, 26.480, 28.180, 29.520, 29.960, 30.440, 31.880, 32.460, 33.120, 35.800, 36.460, 37.100, 37.680, 38.860, 40.620 and 43.

480.

3. The tanshinone sodium II crystal form according to claim 1, characterized in that: The tanshinone sodium II crystal form has an XRPD spectrum that is substantially the same as Figure 1 of the accompanying drawing of the specification.

4. The method for preparing the Danshensu Sodium II crystal form according to any one of claims 1 to 3, comprising: 1) Take Danshensu Sodium API, add water for the first time, heat and stir to dissolve; Then add the organic solvent for the second time and continue stirring to dissolve; 2) Keep stirring at room temperature until a large amount of crystals are precipitated, filter and dry to obtain sodium danshensu crystals; The mass volume ratio of the sodium danshensu raw material and water in step 1) is 1 g: 2 mL; The organic solvent in step 1) is selected from ethanol; wherein the amount of the organic solvent used is twice the amount of water used; The temperature of heating, stirring and dissolving in step 1) is controlled at 10-70°C; The temperature of adding the organic solvent for the second time in step 1) and continuing to stir and dissolve is controlled at 0-50°C.

5. The method for preparing the Danshensu Sodium II crystal form according to any one of claims 1 to 3, comprising: Take 10g of sodium danshensu raw material, add 30mL of ethanol, 5mL of n-butanol, and then add 40mL of purified water, heat to 60℃ to dissolve, keep warm and stir for 1 hour, then slowly cool to 12℃, stir and crystallize for 12h, a large amount of crystals precipitate, filter; place the filter cake in 20mL of ethanol, stir for 8h, and filter; the filter cake is vacuum dried at 60℃ to obtain sodium danshensu crystals.

6. The preparation method according to claim 4, characterized in that: The stirring time at room temperature is controlled to be 12 to 48 hours.

7. The preparation method according to claim 4, characterized in that: The drying treatment is specifically performed by drying under reduced pressure in a vacuum oven at 80° C. for 24 hours or by drying under reduced pressure using an oil pump for 12 hours.

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