Gastrodin anhydrous crystalline type and preparation method thereof

Abstract

The present invention discloses a high-purity anhydrous gastrodin, a crystal form, and a method for preparing the crystal form and an application thereof. The X-ray powder diffraction spectrum obtained by using Cu-Kα ray measurement shows that this crystal form is discovered for the first time. The present invention enriches the crystal form research of gastrodin and broadens the application scenarios of gastrodin. The preparation process of the present invention is simple, and the obtained crystal form compound has high purity, low hygroscopicity, and good relative stability.

Description

Technical Field

[0001] The invention relates to the technical field of pharmaceutical chemistry, and in particular to an anhydrous crystalline form of gastrodin and a preparation method thereof. Background Art

[0002] Gastrodia elata Bl is the dried tuber of the orchid plant Gastrodia elata, also known as red arrow, mother, ghost Duyou, magic grass, single shake Zhi, red arrow fat, Dingfeng grass, Heli grass, single shake, automatic grass, water potato. It is dug from the beginning of winter to before Qingming of the following year (winter hemp is better), and is a famous Chinese medicinal material. It has been used as medicine as early as more than 2,000 years ago, and the ones produced in Zhaotong, Yunnan are the best. It is rich in gastrodin, vanillin, protein, amino acids, and trace elements. It is pungent, warm, and non-toxic. The Chinese Pharmacopoeia records its functions and indications as: anti-epileptic, anti-palpitation, anti-rheumatic; sedative, antispasmodic, analgesic, tonic, calming the liver and calming the wind. Functions and indications: calming the liver, calming the wind and stopping spasms. It is used for headache, dizziness, numbness of limbs, epilepsy, convulsions, and tetanus. Dizziness, neurasthenia, wind-cold dampness, infantile convulsions, etc. Clinical application has proved that it has significant therapeutic effects on vascular neurosis headache, sequelae of concussion, etc. Pharmacological studies have shown that the main active ingredient of Gastrodia elata is gastrodin. The chemical name is: 4-hydroxymethylbenzene-β-D-pyranoglucoside hemihydrate, molecular formula C 13 H 19 O8, molecular weight 295.38.

[0003] The commonly used dosage forms of gastrodin in clinical use include injection, powder injection, tablets, capsules, etc. The research reports on the crystal form of gastrodin are as follows:

[0004] Patent CN 103224539 B mentions a crystal form of gastrodin: gastrodin is dissolved in ethanol / methanol or a mixed solvent of the two at room temperature, filtered through carbon, anti-solvent chloroform / ether is added dropwise, crystallized at low temperature, and dried to obtain the crystal form.

[0005] Patent CN 103788148 B mentions a gastrodin crystal form: gastrodin is dissolved in acetone / DMSO or a mixed solvent of the two at room temperature, filtered through carbon, anti-solvent chloroform is added dropwise, crystallized, and dried to obtain the crystal form.

[0006] Patent CN 104447904 A mentions a crystal form of gastrodin: gastrodin is dissolved in ethanol / methanol or a mixed solvent of the two at room temperature, filtered through carbon, anti-solvent ethyl acetate is added dropwise, crystallized at low temperature, and dried to obtain the crystal form.

[0007] The above-mentioned reports on the crystalline compounds of gastrodin are all hemihydrates. Summary of the invention

[0008] In view of this, the technical problem to be solved by the present invention is to provide a crystalline form of gastrodin without crystals and a preparation method thereof.

[0009] In order to solve the above technical problems, the present invention provides a crystalline form of gastrodin without crystals, and the X-ray powder diffraction pattern has a diffraction peak at 2θ±0.2°, and the 2θ is 10.522°, 13.843°, 14.504°, 18.526°, 21.426°, 22.203°, 23.200°, 23.906° and 24.383°.

[0010] Further, the 2θ is 10.522°, 12.142°, 13.843°, 14.504°, 15.003°, 15.738°, 17.587°, 18.041°, 18.526°, 19.431°, 20.370°, 20.624°, 21.078°, 21.426°, 22.203°, 23.200°, 23.906°, 24.383°, 25.962°, 27.141°, 27. 647°, 27.842°, 28.445°, 28.647°, 29.050°, 30.333°, 30.961°, 31.802°, 32.232°, 32.944°, 33.601°, 34.100°, 34.439°, 34.805°, 35.425°, 35.581°, 36.052°, 36.285°, 37.103°, 37.504°, 37.706° and 39.660°.

[0011] The structural formula of gastrodin is shown in Formula I:

[0012]

[0013] Specifically, the detailed parameters of the X-ray diffraction of the anhydrous crystalline form of gastrodin described in the present invention are as follows: Figure 1 shown.

[0014] The X-ray powder diffraction pattern obtained by Cu-Ka ray measurement is as follows: Figure 2 shown.

[0015] The present invention names the newly prepared gastrodin anhydrous crystalline form as Form 2.

[0016] In the present invention, the 2θ value of the X-ray powder diffraction pattern may vary slightly between machines or samples, and its value may differ by about 0.2 units, or by about 0.1 units, so the quoted values ​​cannot be interpreted as absolute values. It should also be understood that the size of the peak height may also differ by about 5 units, or by about 4 units, or by about 3 units, or by about 2 units, or by about 1 unit, so the XRPD trace intensity included in the present invention is illustrative and is not intended to be used for absolute comparison.

[0017] The present invention provides a method for preparing the above-mentioned anhydrous crystalline form of gastrodin, comprising the following steps:

[0018] The hemihydrate of gastrodin Form A crystal form is dissolved in a solvent, an anti-solvent is added for crystallization, and the solid is separated and dried to obtain the anhydrous crystalline form of gastrodin Form 2.

[0019] Preferably, the solvent is acetone, and the anti-solvent is chloroform.

[0020] In the present invention, the Form A crystal hemihydrate is prepared according to the following method:

[0021] Gastrodin is dissolved in ethanol, methanol or a mixed solvent of the two at room temperature, filtered through carbon, anti-solvent ethyl acetate is added dropwise, crystallized at low temperature, and dried to obtain gastrodin Form A crystal hemihydrate.

[0022] For details, please refer to the Chinese patent application number 201410379348.6.

[0023] The present invention also provides a method for preparing anhydrous crystalline gastrodin, comprising the following steps:

[0024] The gastrodin is stirred at low temperature in a solvent, and the solid is separated and dried to obtain a gastrodin anhydrous crystalline form.

[0025] The solvent is preferably acetonitrile, methyl tert-butyl ether or ethyl acetate.

[0026] Alternatively, the solution can be dissolved in tetrahydrofuran and chloroform can be added as an anti-solvent.

[0027] Alternatively, the product may be dissolved in trifluoroethanol, nitromethane may be added, and the product may be left in an open state until the solvent evaporates to dryness, thereby obtaining anhydrous crystalline form of gastrodin.

[0028] The Form A crystal hemihydrate is prepared according to the above method.

[0029] In some specific embodiments of the present invention, the above preparation method is specifically:

[0030] Gastrodin is stirred in acetonitrile at low temperature. If it does not dissolve, stir overnight at -10 to 10°C, preferably 4°C. After the solid is separated, dry to obtain gastrodin anhydrous crystalline Form 2.

[0031] The temperature of the low-temperature stirring is preferably 50 to 70°C, more preferably 60°C.

[0032] The drying condition is preferably room temperature vacuum drying, and the drying time is preferably 3 to 8 hours.

[0033] In some specific embodiments of the present invention, the above preparation method is specifically:

[0034] The slurry of gastrodin is stirred in methyl tert-butyl ether at -10 to 10°C, preferably at 4°C. The stirring time is preferably 24 to 90 hours, preferably 48 hours. The solid is separated and dried to obtain gastrodin anhydrous crystalline form Form 2.

[0035] The drying condition is preferably room temperature vacuum drying, and the drying time is preferably 3 to 8 hours.

[0036] In some specific embodiments of the present invention, the above preparation method is specifically:

[0037] Gastrodin was dissolved in tetrahydrofuran, and chloroform was added for crystallization. The solid was separated and dried to obtain gastrodin anhydrous crystalline Form 2.

[0038] In some specific embodiments of the present invention, the above preparation method is specifically:

[0039] Gastrodin is allowed to stand in an ethyl acetate atmosphere, preferably for 2 to 3 days, to obtain anhydrous crystalline form of gastrodin Form 2.

[0040] In some specific embodiments of the present invention, the above preparation method is specifically:

[0041] Gastrodin is dissolved in trifluoroethanol, preferably by ultrasonic dissolution, and then nitromethane is added. After filtering, the filtrate is placed at 30-50° C., preferably 40° C., and evaporated to dryness in an open air. Regular large-grained crystals are precipitated at the bottom to obtain gastrodin anhydrous crystalline Form 2.

[0042] The present invention performs DSC and TGA tests on the prepared gastrodin anhydrous crystalline form Form 2, and the DSC spectrum results show that the melting point of the gastrodin anhydrous crystalline form is 140°C.

[0043] TGA results show that the anhydrous crystalline form of gastrodin Form 2 has a slow weight loss of 0.2% before 120°C, is an anhydrous substance, and has a decomposition temperature of 279°C.

[0044] The DVS / isothermal adsorption curve shows that the weight change of the anhydrous crystalline compound of gastrodin in the range of 0% RH to 80% RH is 0.6%, which is slightly hygroscopic.

[0045] The gastrodin anhydrous crystalline compound is placed under dry conditions at room temperature for 14 days without changing its crystal form.

[0046] When left exposed at room temperature ≤33% RH for 7 days, the crystal form remains unchanged.

[0047] The mixture was left exposed at room temperature (75% RH) for 1 day and then transformed into Form A crystal.

[0048] The present invention provides the use of the above-mentioned anhydrous crystalline gastrodin or the anhydrous crystalline gastrodin prepared by the above-mentioned preparation method in preparing drugs for preventing, alleviating or treating vascular neurosis headache and sequelae of concussion.

[0049] The present invention provides a pharmaceutical preparation comprising the above-mentioned anhydrous crystalline form of gastrodin, or the anhydrous crystalline form of gastrodin prepared by the above-mentioned preparation method and pharmaceutically acceptable excipients.

[0050] The dosage form of the pharmaceutical preparation can be tablets, hard capsules, soft capsules, dry suspensions, dropping pills, micropills, etc.

[0051] The present invention provides a pharmaceutical composition, comprising the above-mentioned anhydrous crystalline gastrodin, or the anhydrous crystalline gastrodin prepared by the above-mentioned preparation method and other drugs for treating vascular neurotic headache and sequelae of concussion.

[0052] The present invention provides a new anhydrous crystalline form of gastrodin. The X-ray powder diffraction spectrum measured using Cu-Kα rays shows that this crystalline form is discovered for the first time. The present invention enriches the crystalline form research of gastrodin and broadens the application scenarios of gastrodin. The preparation process of the present invention is simple, and the obtained crystalline compound has high purity, low hygroscopicity and good relative stability.

[0053] It should be noted that the formation of a crystal form is affected by many factors. Even a small change in any factor such as reaction temperature, time, stirring speed, reactant concentration, crystallization condition control, etc. may produce unexpected changes. BRIEF DESCRIPTION OF THE DRAWINGS

[0054] Figure 1 This is an X-ray diffraction parameter diagram of the anhydrous crystalline form of gastrodin Form 2 obtained in Example 1 of the present invention;

[0055] Figure 2 The X-ray powder diffraction pattern of the anhydrous crystalline form of gastrodin Form 2 prepared in Example 1 of the present invention, wherein the ordinate represents the diffraction intensity expressed in counts / second (cps), and the abscissa represents the diffraction angle 2θ expressed in degrees;

[0056] Figure 3 is the X-ray powder diffraction pattern of gastrodin Form A crystal hemihydrate;

[0057] Figure 4 This is the DSC graph of the anhydrous crystalline form of gastrodin Form 2 obtained in Example 1 of the present invention;

[0058] Figure 5 This is the TGA chart of the anhydrous crystalline form of gastrodin Form 2 obtained in Example 1 of the present invention;

[0059] Figure 6 The isothermal adsorption curve of the anhydrous crystalline form of gastrodin Form 2 prepared in Example 1 of the present invention;

[0060] Figure 7 This is the DVS spectrum of the anhydrous crystalline form of gastrodin Form 2 obtained in Example 1 of the present invention;

[0061] Figure 8 This is a PLM morphology diagram of the gastrodin anhydrous crystalline Form 2 obtained in Example 1 of the present invention;

[0062] Fig. 9 This is the XRPD diagram of the stability of the gastrodin anhydrous crystalline Form 2 sealed and dried at room temperature obtained in Example 1 of the present invention;

[0063] Fig.10 The XRPD diagrams of the anhydrous crystalline form of gastrodin Form 2 prepared in Example 1 of the present invention at room temperature ≤ 33% RH, 44% RH, and 75% RH respectively;

[0064] Fig.11 It is the Form liquid phase diagram. DETAILED DESCRIPTION

[0065] In order to further illustrate the present invention, the anhydrous crystalline form of gastrodin provided by the present invention and the preparation method thereof are described in detail below in conjunction with examples.

[0066] In the following examples, unless otherwise stated, the test methods are generally carried out according to conventional conditions or conditions recommended by the manufacturers.

[0067] The X-ray powder diffraction pattern of the present invention was collected on a Bruker D8 Advance Diffractometer X-ray powder diffractometer. The method parameters of the X-ray powder diffraction of the present invention are as follows:

[0068] X-ray reflection parameters: Cu-Ka

[0069] Voltage: 40 kilovolts (kV)

[0070] Current: 40 milliamperes (mA)

[0071] Divergence slit: 1 / 32°

[0072] Anti-scatter slit: 1 / 16°

[0073] Scanning range: from 3.0 to 40.0 degrees

[0074] Sampling step: 0.02 degrees

[0075] Measurement time per step: 0.2 seconds / step

[0076] The differential scanning calorimetry analysis diagram of the present invention is collected on a DSC TA Instruments Q200 differential scanning calorimeter.

[0077] The method parameters of differential scanning calorimetry analysis described in the present invention are as follows:

[0078] Temperature range / ℃:0℃-250℃

[0079] Scanning rate / ℃ / min:10℃ / min

[0080] Protective gas: Nitrogen, 50 ml / min

[0081] The thermogravimetric analysis diagram of the present invention was collected on a TA Instruments Q500TGA thermogravimetric analyzer.

[0082] The method parameters of the thermogravimetric analysis described in the present invention are as follows:

[0083] Temperature range / ℃:10-350℃

[0084] Scanning rate / ℃ / min:10℃ / min

[0085] Protective gas: Nitrogen, 40 ml / min

[0086] The dynamic moisture adsorption instrument, analytical balance, and constant temperature and humidity chamber used in the present invention are shown in Table 1 below:

[0087] Table 1 Parameters of the dynamic moisture adsorption instrument, analytical balance, and constant temperature and humidity chamber used in the present invention

[0088]

[0089]

[0090] The following preparation method of Form A crystalline gastrodin hemihydrate refers to Chinese patent application number 201410379348.6.

[0091] Example 1 Preparation of Form 2 Crystalline Form of Gastrodin

[0092] Take about 200 mg of gastrodin sample, add 20 mL of acetonitrile, stir at 60°C for about 60 minutes, if it is not dissolved, transfer to 4°C and stir overnight, centrifuge, and vacuum dry at room temperature for about 5 hours to obtain Form 2 crystal sample.

[0093] Example 2 Preparation of Gastrodin Form 2 Crystalline Form

[0094] Take about 200 mg of gastrodin sample, add 5 mL of methyl tert-butyl ether, stir the slurry at 4°C for 2 days, centrifuge, and vacuum dry at room temperature overnight to obtain Form 2 crystal sample.

[0095] Example 3 Preparation of Form 2 Crystalline Form of Gastrodin

[0096] Take about 20 mg of gastrodin sample, add 1 mL of tetrahydrofuran to dissolve, filter, add 3 mL of chloroform dropwise under stirring at room temperature to precipitate solid, continue stirring for 30 minutes to 60 minutes, centrifuge, and vacuum dry at room temperature overnight to obtain Form 2 crystal sample.

[0097] Example 4 Preparation of Gastrodin Form 2 Crystalline Form

[0098] About 20 mg of gastrodin sample was taken, placed in a centrifuge tube, and placed in an ethyl acetate solvent atmosphere at room temperature for 3 days to obtain a Form 2 crystal sample.

[0099] Example 5 Preparation of Form 2 Crystalline Form of Gastrodin

[0100] Take about 100 mg of gastrodin sample, add 5 mL of trifluoroethanol, sonicate to dissolve, then add 2 mL of nitromethane, filter without precipitation, and place the filtrate at 40°C to evaporate to dryness. Regular large-grained crystals precipitate at the bottom to obtain Form 2 crystal sample.

[0101] Example 6 Preparation of Gastrodin Form 2 Crystalline Form

[0102] Take about 20 mg of gastrodin Form A crystal sample, add 3 mL of acetone, filter the solution, add 6 mL of chloroform dropwise at room temperature with stirring to precipitate solid, continue stirring for 30 min-60 min, centrifuge, and vacuum dry at room temperature overnight to obtain Form 2 crystal sample.

[0103] Experiment 1: X-ray powder diffraction detection

[0104] The X-ray diffraction test of the gastrodin Form 2 crystal prepared in Example 1 was performed, and the results were as follows: Figure 2 shown.

[0105] The raw material gastrodin Form A crystal hemihydrate was tested by X-ray diffraction, and the results are as follows Figure 3 shown.

[0106] This indicates that sample 2 is an anhydrous crystalline form and has a different X-ray diffraction pattern from the Form A crystalline form.

[0107] X-ray powder diffraction test was performed on samples 2-6, and the results were consistent with Figure 2 resemblance.

[0108] Experiment 2: Differential Scanning Calorimetry and Thermogravimetric Analysis

[0109] Sample 1 was taken for DSC and TGA testing, and the results were as follows: Figure 4 , Figure 5 As shown, the results show that the melting point of sample 1 is 140°C, there is a slow weight loss of 0.2% before 120°C, it is anhydrous, and the decomposition temperature is 279°C.

[0110] DSC and TGA tests were performed on samples 2 to 6, and the results were consistent with Figure 4 , Figure 5 resemblance.

[0111] Experiment 3: Dynamic moisture adsorption detection

[0112] Sample 1 was taken for isothermal adsorption test and DVS analysis experiment. The isothermal adsorption curve is shown in Figure 6 As shown, the DVS spectrum is as follows Figure 7 shown.

[0113] The results showed that the weight change of sample 1 in the range of 0% RH to 80% RH was 0.6%, indicating slight hygroscopicity.

[0114] Isothermal adsorption tests and DVS analysis experiments were performed on samples 2-6, and the results were similar.

[0115] Experiment 4: PLM

[0116] The crystal morphology was observed using a polarizing microscope. The PLM morphology is shown in the figure below. Figure 7 shown.

[0117] Experiment 5: Accelerated stability evaluation experiment

[0118] Take an appropriate amount of sample 1, place it in a sealed dry environment at room temperature for 14 days, place it in an open environment at room temperature ≤ 33% RH for 7 days, and place it in an open environment at 75% RH for 1 day, and conduct an accelerated stability test. Use XRPD to characterize the sample and detect its crystal form. The test results are as follows: Figure 8 , 9 shown.

[0119] The test results show that the crystal form of sample 1 remains unchanged after being placed in dry conditions at room temperature for 14 days.

[0120] When left exposed at room temperature ≤33% RH for 7 days, the crystal form remains unchanged.

[0121] When left exposed at room temperature (44% RH) for 7 days, the crystal form remained essentially unchanged.

[0122] The mixture was left exposed at room temperature (75% RH) for 1 day and then transformed into Form A crystal.

[0123] Samples 2-4 were subjected to accelerated stability evaluation tests with similar results.

[0124] This indicates that the gastrodin crystal form Form 2 prepared by the present invention has good stability.

[0125] Experiment 6:

[0126] This experimental example compares the hygroscopicity of the gastrodin compound provided by the present invention with the hygroscopicity of the gastrodin compound with better hygroscopicity effect reported in existing patents.

[0127] The patent number CN104628796B reports that the hygroscopicity of the crystal form of this patent is only 0.2% (30°C, 75%).

[0128] Patent No. CN105524127B reports that the hygroscopicity of the crystal form of this patent is 0.7%.

[0129] The samples were prepared according to the methods reported in CN 104628796B and CN105524127B.

[0130] Test method: Under the condition of 80% humidity and room temperature, 10g of each sample was placed on an electronic balance and the weight was recorded regularly to detect the degree of moisture absorption. The results are shown in Table 2.

[0131] Table 2 Hygroscopicity results

[0132]

[0133]

[0134] Experiment 7:

[0135] This experimental example investigates the stability comparison between the gastrodin crystal form provided by the present invention and the gastrodin crystal form in the prior art. This test is conducted in accordance with the guidelines for drug stability tests in Appendix XIXC of Part II of the Chinese Pharmacopoeia 2015 edition, and an accelerated test is performed.

[0136] The experiment examined the stability of gastrodin crystals by their purity. The results of the accelerated test are shown in Table 3 below:

[0137] Table 3 Stability accelerated test results

[0138]

[0139]

[0140] It can be seen from the above examples that the gastrodin crystal form Form 2 provided by the present invention has higher stability.

[0141] The above embodiments are only used to help understand the method and core idea of ​​the present invention. It should be noted that, for those skilled in the art, several improvements and modifications can be made to the present invention without departing from the principles of the present invention, and these improvements and modifications also fall within the scope of protection of the claims of the present invention.

Claims

1. A crystalline form of gastrodin without crystals, characterized in that: The X-ray powder diffraction pattern has diffraction peaks at 2θ±0.2°, and the 2θ is 10.522°, 13.843°, 14.504°, 18.526°, 21.426°, 22.203°, 23.200°, 23.906° and 24.383°; The preparation method of the anhydrous crystalline form of gastrodin comprises the following steps: The Form A crystal hemihydrate sample is dissolved in a solvent, an anti-solvent is added for crystallization, and the solid is separated and dried to obtain anhydrous gastrodin crystals; the solvent is acetone, and the anti-solvent is chloroform; or stirring gastrodin in a solvent at low temperature, separating the solid and drying it to obtain gastrodin anhydrous crystalline form; the solvent is acetonitrile; the temperature of the low temperature stirring is 50-70° C.; The Form A crystalline hemihydrate is prepared according to the following method: Gastrodin is dissolved in ethanol, methanol or a mixed solvent of the two at room temperature, filtered through carbon, anti-solvent ethyl acetate is added dropwise, crystallized at low temperature, and dried to obtain gastrodin Form A crystal hemihydrate.

2. The anhydrous crystalline form of gastrodin according to claim 1, characterized in that: The 2θ is 10.522°, 12.142°, 13.843°, 14.504°, 15.003°, 15.738°, 17.587°, 18.041°, 18.526°, 19.431°, 20.370°, 20.624°, 21.078°, 21.426°, 22.203°, 23.200°, 23.906°, 24.383°, 25.962°, 27.141°, 27.64 7°, 27.842°, 28.445°, 28.647°, 29.050°, 30.333°, 30.961°, 31.802°, 32.232°, 32.944°, 33.601°, 34.100°, 34.439°, 34.805°, 35.425°, 35.581°, 36.052°, 36.285°, 37.103°, 37.504°, 37.706° and 39.660°.

3. The anhydrous crystalline form of gastrodin according to claim 1, characterized in that: The X-ray powder diffraction pattern is shown in FIG2 .

4. The anhydrous crystalline form of gastrodin according to claim 1, characterized in that: The melting point of the anhydrous crystalline form of gastrodin is 140°C; the decomposition temperature of the anhydrous crystalline form of gastrodin is 279°C.

5. The method for preparing the anhydrous crystalline form of gastrodin according to any one of claims 1 to 4, comprising the following steps: The Form A crystal hemihydrate sample is dissolved in a solvent, an anti-solvent is added for crystallization, and the solid is separated and dried to obtain an anhydrous form of gastrodin; The solvent is acetone, and the anti-solvent is chloroform.

6. The method for preparing the anhydrous crystalline form of gastrodin according to any one of claims 1 to 4, comprising the following steps: Gastrodin is stirred at low temperature in a solvent, and the solid is separated and dried to obtain anhydrous crystalline gastrodin; The solvent is acetonitrile.

7. Use of the anhydrous crystalline gastrodin as described in any one of claims 1 to 4, or the anhydrous crystalline gastrodin prepared by the preparation method described in any one of claims 5 to 6 in the preparation of drugs for preventing, alleviating or treating vascular neurosis headache and sequelae of concussion.

8. A pharmaceutical preparation, characterized in that The invention comprises the anhydrous crystalline form of gastrodin as described in any one of claims 1 to 4, or the anhydrous crystalline form of gastrodin prepared by the preparation method as described in any one of claims 5 to 6, and pharmaceutically acceptable excipients.

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