Crystal of solvate of episesamin with n-butanol

Abstract

This application discloses a solvate crystal of citric acid and n-butanol, which has crystal type II, wherein the molar ratio of citric acid to n-butanol is 1:2, and the crystal has higher solubility and bioavailability.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology. Specifically, it relates to solvate crystals of cimicifugain and n-butanol, their preparation methods, compositions, and uses. Background Technology

[0002] The chemical name of the verbascoside is (E)-(2R,3R,4R,5R,6R)-6-(3,4-dihydroxyphenylethoxy)-5-hydroxy-2-(hydroxymethyl)-4-(((2S,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-3-yl 3-(3,4-dihydroxyphenyl)acrylate, with the molecular formula C 29 H 36 O 15 The structural formula is as follows:

[0003]

[0004] Citrulline-like glycosides are glycoside compounds, also known as verbascoside or ergosterol. Physicochemically, they appear as off-white or pale yellow crystalline powders, slightly soluble in methanol or ethanol, soluble in dimethyl sulfoxide, and insoluble in water. They possess antitumor, anti-anxiety, and antidepressant activities, can improve memory impairment, and are also effective in treating inflammation. [1-9] .

[0005] Currently, commercially available cimicifuga glycosides are available as cimicifuga glycoside powder (defined as crystal type I). However, due to their poor water solubility and poor absorption in the stomach, cimicifuga glycosides have poor oral efficacy, resulting in low bioavailability and limiting their effects in the body. Therefore, taking effective measures to improve the water solubility of cimicifuga glycosides to better exert their therapeutic effects is of great significance. Summary of the Invention

[0006] One or more embodiments of this application provide a solvate crystal of cimicifugain and n-butanol, wherein the molar ratio of cimicifugain to n-butanol is 1:2, which has a crystal type II. When irradiated with CuKα and expressed in 2θ angles, the powder X-ray diffraction pattern of the crystal type II has the following characteristic peaks: 4.9±0.2°, 7.0±0.2°, 8.2±0.2°, 8.9±0.2°, 11.5±0.2°, 12.6±0.2°, 17.0±0.2°, 19.9±0.2°, 21.0±0.2°, 21.4±0.2°, 25.4±0.2°.

[0007] In one or more embodiments, CuK is used. αRadiation, expressed at a 2θ angle, the powder X-ray diffraction pattern of the type II crystal has the following characteristic peaks and peak area percentages:

[0008]

[0009] In one or more embodiments, CuK is used. α Radiation, expressed in 2θ angles, and the powder X-ray diffraction pattern of the type II crystal has the following characteristic peaks: 4.9±0.2°, 7.0±0.2°, 8.2±0.2°, 8.9±0.2°, 9.9±0.2°, 11.5±0.2°, 12.6±0.2°, 13.3±0.2°, 14.0±0.2°, 14.4±0.2°, 14.8±0.2°, 15.3±0.2°, 17.0±0.2°, 17.5±0.2°, 17.9±0.2°. 18.6±0.2°, 19.9±0.2°, 20.3±0.2°, 20.8±0.2°, 21.0±0.2°, 21.4±0.2°, 21.6±0.2°, 22.1±0.2°, 22.9±0.2°, 23.2±0.2°, 24.1±0.2°, 24.6±0.2°, 25.4±0.2°, 27.6±0.2°, 29.7±0.2°, 30.0±0.2°, 30.9±0.2°, 45.1±0.2°.

[0010] In one or more embodiments, CuK is used. α Radiation, expressed at a 2θ angle, the powder X-ray diffraction pattern of the type II crystal has the following characteristic peaks and peak area percentages:

[0011]

[0012]

[0013] In one or more embodiments, CuK is used. α Radiation is expressed in 2θ angles, and the powder X-ray diffraction pattern of the type II crystal is as follows: Figure 1 As shown.

[0014] In one or more embodiments, when analyzed using attenuated total reflectance Fourier transform infrared spectroscopy, the infrared spectrum of the crystal type II is at 3263±2 cm⁻¹. -1 2953±2cm -1 2886±2cm -1 1693±2cm -1 1636±2cm -1 1607±2cm -1 1530±2cm -11444±2cm -1 1381±2cm -1 1340±2cm -1 1308±2cm -1 1286±2cm -1 1272±2cm -1 1252±2cm -1 1204±2cm -1 1180±2cm -1 1167±2cm -1 1151±2cm -1 1140±2cm -1 1115±2cm -1 1085±2cm - 1. 1072±2cm -1 1059±2cm -1 1040±2cm -1 1021±2cm -1 1008±2cm -1 976±2cm -1 906±2cm -1 871±2cm -1 850±2cm -1 829±2cm -1 816±2cm -1 807±2cm - 1. 784±2cm -1 739±2cm -1 714±2cm -1 There are characteristic peaks in the infrared spectrum at that location.

[0015] In one or more embodiments, the infrared spectrum of the crystal type II is as follows: Figure 3 As shown.

[0016] In one or more embodiments, the differential scanning calorimeter of the crystal type II (see...) Figure 4 In the study, two endothermic peaks were observed at 106±3℃ and 133±3℃.

[0017] In one or more embodiments, the differential scanning calorimeter of the crystal type II (see...) Figure 4 In the range of 30 to 200℃, the heating rate is 10℃ / min.

[0018] One or more embodiments of this application provide a method for preparing the solvate crystal of cimicifugain and n-butanol of this application, which is a suspension stirring method, a solvent evaporation crystallization method, or a liquid addition grinding method.

[0019] In one or more embodiments, the suspension stirring method includes: adding n-butanol as a single solvent or a mixture of n-butanol and an organic solvent to the cimicifuga glycoside, stirring at 300-500 r / min for 72-240 h at a temperature of 20°C-45°C, drying the product, and obtaining crystals with crystal type II.

[0020] In one or more embodiments, the solvent evaporation crystallization method includes: dissolving citric acid in n-butanol as a single solvent or a mixed solvent of n-butanol and water in a volume ratio of 4:1 at a temperature of 15°C to 60°C, wherein the mass ratio of citric acid to organic solvent is 0.1 mg / mL to 20 mg / mL, stirring for 2 to 24 hours, placing the solution at a temperature of 20°C to 40°C, and slowly evaporating the solvent for 14 to 30 days at a relative humidity of 10% to 90%, until the solvent has evaporated completely, thereby obtaining crystals of crystal type II.

[0021] In one or more embodiments, the liquid grinding method includes: (1) placing the physcinoside in a mortar and grinding it evenly; (2) taking the substance obtained in step (1) and grinding it in a clockwise direction, and adding n-butanol or a mixture of n-butanol and organic solvent every 15 minutes, with each addition of solvent being 2 mL / g to 10 mL / g, and grinding for 12 to 48 hours; (3) drying the product obtained in step (2) to obtain crystals with crystal type II.

[0022] In one or more embodiments, the organic solvent is selected from one or more of methanol, ethanol, tetrahydrofuran, acetone, acetonitrile, dioxane, n-hexane, and cyclohexane.

[0023] In one or more embodiments, the volume ratio between any two organic solvents is 1:10 to 10:1, for example 1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1.

[0024] One or more embodiments of this application provide a solid mixture comprising crystals of the solvate of the present application of cimicifugain and n-butanol, wherein the amount of the crystals is 1-99.9 wt%, for example 10-99.9 wt%, 50-99.9 wt%, or 85-99.9 wt%, or for example 10 wt%, 20 wt%, 30 wt%, 40 wt%, 50 wt%, 60 wt%, 70 wt%, 80 wt%, or 90 wt%, based on the weight of the solid mixture.

[0025] One or more embodiments of this application provide pharmaceutical compositions comprising a therapeutically effective amount of a solvate crystal of the present application’s cimicifugain and n-butanol or a solid mixture of the present application and a pharmaceutically acceptable carrier.

[0026] In one or more embodiments, the daily dose of the pharmaceutical composition is 10 to 1000 mg, for example 100 mg, 500 mg, or 900 mg.

[0027] In one or more embodiments, the pharmaceutical composition is prepared as a tablet, capsule, pill, injectable formulation, powder for injection, granule, powder, micro-pellet, drop, suppository, film, patch, aerosol, or spray.

[0028] In one or more embodiments, the pharmaceutical composition is prepared as a sustained-release formulation or a controlled-release formulation.

[0029] One or more embodiments of this application provide the use of the solvate crystals of the present application and n-butanol, the solid mixtures of the present application, or the pharmaceutical compositions of the present application in the preparation of medicaments for the prevention and / or treatment of inflammation, anxiety, depression, tumors, and neurodegenerative diseases.

[0030] One or more embodiments of this application provide the use of the solvate crystals of the present application of cimicifugain and n-butanol, the solid mixtures of the present application, or the pharmaceutical compositions of the present application in the preparation of medicaments for immunomodulation.

[0031] One or more embodiments of this application provide cimicifugoside crystals, wherein cimicifugoside forms a solvate with n-butanol in a 1:2 molar ratio.

[0032] In one or more embodiments, the thermogravimetric spectrum of the crystal type II ( Figure 5 Two weight loss peaks were observed in the temperature range of 80-130℃, with a weight loss of 15% to 23%.

[0033] In one or more embodiments, the thermogravimetric spectrum of the crystal type II ( Figure 5 Within the temperature range of 30 to 500℃, the heating rate is 5℃ / min.

[0034] In one or more embodiments, the solvent evaporation crystallization method involves adding 5 to 100 mL of organic solvent (e.g., 20 mL) per gram of cimicifugoside sample, crystallizing for 0.1 to 10 hours (e.g., 5 hours), drying at 25-60°C (e.g., 40°C), and drying for 2 to 10 hours (e.g., 6 hours).

[0035] In one or more embodiments, the molecular formula of the solvate crystal of cimicifugain and n-butanol is C29 H 36 O 15 .

[0036] In one or more embodiments, the solvate crystals of citric acid and n-butanol of this application have better solubility than citric acid crystal type I.

[0037] In one or more embodiments, a mixed solid substance or pharmaceutical composition comprising any non-zero proportion of solvate crystals of citronellol and n-butanol is provided.

[0038] In one or more embodiments, the solvate crystals of the present application of cimicifugoside and n-butanol are readily released during the treatment of diseases, producing unexpected clinical effects.

[0039] Morphological characteristics of the solvate crystals of citric acid-like glycosides and n-butanol (also referred to as "citric acid-like glycoside crystals" in this paper).

[0040] This application relates to a cimicifugoside crystal, wherein cimicifugoside and n-butanol form a solvate in a 1:2 molar ratio.

[0041] This application describes the cimicifugoside crystals, when analyzed using powder X-ray diffraction with CuK... α Under radiation experimental conditions, the diffraction peak positions are: 2 - Theta value (°) or d value. The relative intensities of diffraction peaks (peak height (Height%) or peak area (Area%)) are shown in Table 1. Figure 1 As shown in the image.

[0042] The X-ray diffraction pattern and data of the crystalline type I of cimicifuga glycoside are shown in Table 2. Figure 2 The powder X-ray diffraction patterns of the crystal type II and crystal type I of cimicifugaside crystals show significant differences in the number, position, intensity, and topology of diffraction peaks, indicating that the crystal type II and crystal type I of cimicifugaside crystals in this application are different crystal types.

[0043] Table 1. Powder X-ray diffraction peak values ​​of crystal type II of cimicifugain crystals.

[0044]

[0045]

[0046] The crystal type II of the cimicifugoside crystals described in this application, when analyzed using attenuated total reflectance Fourier transform infrared spectroscopy, showed the following values ​​at 3263, 2953, 2886, 1693, 1636, 1607, 1530, 1444, 1381, 1340, 1308, 1286, 1272, 1252, 1204, 1180, 1167, 1151, 1140, 1115, 1085, 1072, 1059, 1040, 1021, 1008, 976, 906, 871, 850, 829, 816, 807, 784, 739, and 714 cm⁻¹. -1 There is an infrared spectral characteristic peak at this location, with an allowable deviation of ±2cm for the infrared spectral characteristic peak. -1 . ( Figure 3 ).

[0047] The crystal type II of the cimicifugain-like crystals of this application, when analyzed using differential scanning calorimetry (DSC), exhibits two endothermic peaks at 106±3℃ and 133±3℃ in the DSC spectrum when the heating rate is 10℃ per minute. Figure 4 ).

[0048] The crystal type II of the cimicifugain-like crystals described in this application, when analyzed using thermogravimetric analysis, exhibits two weight loss peaks in the TG spectrum within the range of 80-130℃ at a heating rate of 5℃ per minute, with a weight loss of 15%-23%. Figure 5 ).

[0049] Preparation method of cimicifugoside crystals with crystal type II

[0050] The preparation methods of the physcinoside-like crystals in this application include the following methods:

[0051] Suspension stirring method

[0052] Using n-butanol, weigh an appropriate amount of cimicifugain and add 0.5–20 mL of organic solvent. Stir at 300–500 r / min for 72–240 h at 20–45 °C. Dry the product to obtain cimicifugain crystals.

[0053] Solvent evaporation crystallization method

[0054] Using n-butanol as a single solvent or a mixture of n-butanol and water (v / v, 4:1), physcinoside is dissolved in the solvent at a temperature of 15℃ to 60℃. The mass ratio of physcinoside to the volume of the n-butanol single solvent or the n-butanol and water mixture is 0.1 mg / mL to 20 mg / mL. The mixture is stirred for 2 h to 24 h and then placed in an environment with a temperature of 20℃ to 40℃ and a relative humidity of 10% to 90% for slow solvent evaporation for 14 to 30 days. The mixture is then evaporated to dryness with the solvent to obtain physcinoside crystals of crystal type II.

[0055] Liquid-assisted grinding method

[0056] Includes the following steps:

[0057] (1) Place the phytoesin-like glycosides in a mortar and grind them evenly;

[0058] (2) Take the substance obtained in step (1) and grind it in a clockwise direction. Add n-butanol or a mixture of n-butanol and organic solvent every 15 minutes. The amount of solvent added each time is 2 mL / g to 10 mL / g. Grind for 12 to 48 hours.

[0059] (3) The product obtained in step (2) is dried to obtain cimicifugoside crystals of crystal type II.

[0060] Pharmaceutical formulations containing physcinoside crystals of crystal type II, dosage characteristics and pharmaceutical uses

[0061] The pharmaceutical composition of this application contains ephedrine-like crystals and a pharmaceutically acceptable carrier.

[0062] The pharmaceutical composition of this application contains a mixed solid substance of citronellol-like crystals and a pharmaceutically acceptable carrier.

[0063] In the pharmaceutical composition of this application, the daily dosage of phytoescin-like crystals is in the range of 5 to 3000 mg.

[0064] The pharmaceutical compositions involved in this application are prepared as tablets, capsules, pills, injectable preparations, granules, powders, micro-pellets, droplets, suppositories, films, patches, aerosols, sprays, sustained-release preparations, or controlled-release preparations.

[0065] This application relates to the use of ciphorin-like crystals, mixed solid substances containing ciphorin-like crystals, or pharmaceutical compositions in the prevention and treatment of inflammation, tumors, anxiety, depression, neurodegenerative diseases, and immunomodulation.

[0066] This application relates to pharmaceutical compositions using the ephedrine-like crystals of this application as the active ingredient. The pharmaceutical composition can be prepared according to methods known in the art and can be formulated into any dosage form suitable for human or animal use by combining the ephedrine-like crystal component of this application with one or more pharmaceutically acceptable solid or liquid excipients and / or adjuvants. The ephedrine-like crystals of this application are present in the pharmaceutical composition at a concentration ranging from 10% to 90% by weight.

[0067] The phycocyanin crystals described in this application can be administered in unit doses via either the enteric or non-enteric route, primarily orally.

[0068] The preferred dosage form for administration in this application is a solid dosage form. Solid dosage forms can be tablets (including regular tablets, enteric-coated tablets, lozenges, dispersible tablets, chewable tablets, effervescent tablets, orally disintegrating tablets), capsules (including hard capsules, soft capsules, and enteric-coated capsules), granules, powders, microcapsules, droplets, suppositories, films, patches, aerosols, sprays, etc.

[0069] The citronellol crystals described in this application can be formulated into ordinary preparations, as well as sustained-release preparations, controlled-release preparations, targeted preparations, and various microparticle delivery systems.

[0070] In order to formulate the ciphorin crystals of this application into tablets, a wide variety of excipients known in the art can be used, including diluents, binders, wetting agents, disintegrants, lubricants, and flow aids. Diluents can be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, dicalcium phosphate, calcium carbonate, etc.; wetting agents can be water, ethanol, isopropanol, etc.; binders can be starch paste, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, gum arabic paste, gelatin paste, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, polyethylene glycol, etc.; disintegrants can be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, cross-linked polyvinylpyrrolidone, cross-linked sodium carboxymethyl cellulose, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitol fatty acid ester, sodium dodecyl sulfonate, etc.; lubricants and flow aids can be talc, silica, stearate, tartaric acid, liquid paraffin, polyethylene glycol, etc.

[0071] Tablets can also be further processed into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or bilayer and multilayer tablets.

[0072] To formulate the drug delivery unit into capsules, the active ingredient, cimicifugoside crystals as described in this application, can be mixed with a diluent and a flow aid, and the mixture can be directly placed into hard or soft capsules. Alternatively, the active ingredient, cimicifugoside crystals as described in this application, can be first formed into granules or microspheres with a diluent, binder, and disintegrant, and then placed into hard or soft capsules. Various diluents, binders, wetting agents, disintegrants, and flow aids used to prepare cimicifugoside crystal tablets as described in this application can also be used to prepare cimicifugoside crystal capsules as described in this application.

[0073] In addition, colorants, preservatives, flavorings, tasters or other additives may be added to pharmaceutical preparations if necessary.

[0074] To achieve the intended therapeutic purpose and enhance the therapeutic effect, the drug of this application may be administered using any known method of administration.

[0075] The dosage of the pharmaceutical composition of the citronellol crystals described in this application can vary widely depending on the nature and severity of the disease to be prevented or treated, the individual condition of the patient or animal, the route of administration, and the dosage form. The above dosage can be administered as a single unit or divided into several units, depending on the physician's clinical experience and the dosing regimen, including the use of other treatment methods.

[0076] The evodiamine crystals or compositions described herein can be taken alone or in combination with other therapeutic or symptomatic medications. When the evodiamine crystals described herein have a synergistic effect with other therapeutic medications, their dosage should be adjusted according to the actual situation.

[0077] The solubility of the cimicifugoside crystals described in this application is advantageous.

[0078] The equilibrium solubility of the crystalline type II cimicifugain crystals in this application is improved compared to that of the crystalline type I cimicifugain crystals. Figure 6 ). Attached Figure Description

[0079] Figure 1 The powder X-ray diffraction pattern of crystal type II of cimicifugain crystals.

[0080] Figure 2 The powder X-ray diffraction pattern of the crystal type I of cimicifuga glycoside is shown.

[0081] Figure 3 The infrared absorption spectrum of crystal type II of cimicifugoside is shown.

[0082] Figure 4 Differential scanning calorimetry (DSC) spectrum of crystal type II of cimicifugoside.

[0083] Figure 5 Thermogravimetric spectrum of the crystal type II substance of cimicifugoside.

[0084] Figure 6 Equilibrium solubility diagrams for cimicifugoside type II and cimicifugoside type I crystals. Detailed Implementation

[0085] To better illustrate the technical solution of this application, the following embodiments are provided, but this application is not limited thereto.

[0086] Example 1

[0087] Preparation method 1 of cimicifugoside crystals of crystal type II (suspension stirring method)

[0088] Weigh an appropriate amount of cimicifugain and place it in a clean container. Add 0.5–20 mL of organic solvent and stir at a suitable temperature (20℃–45℃) at a speed of 300–500 r / min for an appropriate time (72–240 h). Filter the resulting suspension and dry it under vacuum, by natural filtration or evaporation. Perform powder X-ray diffraction analysis on it. Its diffraction pattern is similar to… Figure 1 The consistency indicates that the obtained sample is a type II cimicifugoside crystal.

[0089] Table 3 Specific examples of preparation method 1

[0090]

[0091] Method 2 for preparing cimicifugoside crystals of crystal type II (liquid-addition grinding method)

[0092] The cimicifugain was placed in a clean mortar and ground evenly. Every 15 minutes, a certain volume of organic solvent was added dropwise, with each addition ranging from 2 mL / g to 80 mL / g. The mixture was ground clockwise for 12–48 hours, and the resulting product was dried. Powder X-ray diffraction analysis was performed, and its diffraction pattern was consistent with... Figure 1 The consistency indicates that the obtained sample is a type II cimicifugoside crystal.

[0093] Table 4 Specific examples of preparation method 2

[0094]

[0095] Method 3 for preparing cimicifugoside crystals of crystal type II (solvent evaporation crystallization method)

[0096] Weigh 10 mg of cimicifugain and add 5 mL of n-butanol or a mixture of n-butanol and water. Stir for 24 h at a suitable temperature (20-50℃), filter, and place in an environment with a temperature of 20-40℃ and a relative humidity of 10%-90% for slow solvent evaporation for 14-30 days to obtain cimicifugain crystals of crystal type II.

[0097] Table 5 Specific examples of preparation method 3

[0098]

[0099] Example 2 Solubility Test

[0100] Compared with the crystal form I of cimicifugain, the crystal II form of this application exhibits improved solubility in hydrochloric acid buffer (pH 1.2), acetate buffer (pH 4.5), phosphate buffer (pH 6.8), and pure water (pH 7.0).

[0101] The equilibrium dissolution concentration of cimicifugoside was used as the evaluation index. The content was determined by high-performance liquid chromatography (HPLC) at a wavelength of 365 nm, and the dissolution concentration was calculated using the external standard method. The data are shown in Table 6.

[0102] Table 6. Equilibrium solubility data of ephedrine crystals of type II and type I.

[0103]

[0104] The experimental data show that the maximum concentration of the crystalline type II cimicifugain crystals of this application that dissolves in four buffer media is significantly improved compared with the crystalline type I cimicifugain crystals. Specifically, the crystalline type II cimicifugain crystals have higher solubility, which improves the rate and extent of bioabsorption of cimicifugain and its bioavailability.

[0105] Example 3 Formulation Preparation

[0106] Preparation method of a drug formulation containing crystalline form I of citric acid glycoside 1 (tablet)

[0107] Preparation method: Using crystalline type II cimicifugoside crystals as the active pharmaceutical ingredient (API) and several excipients as tablet excipients, tablets containing 10–500 mg of drug per tablet were prepared according to a specific ratio. Table 7 shows the tablet formulation ratios.

[0108] Table 7. Formulations for preparing cimicifugoside crystal tablets with crystal type II.

[0109]

[0110] The method for preparing tablet formulations using crystalline type II cimicifugoside crystals as raw materials is as follows: several excipients are mixed evenly with the raw material and directly compressed into tablets; or the excipients are mixed and granulated by dry method, then mixed evenly with the raw material and compressed into tablets.

[0111] Preparation method 2 for combination drug formulations (tablets)

[0112] Preparation method: Pure cimicifugoside crystals of type II crystal form were used as raw materials, and several excipients were used as tablet excipients. The tablets were prepared in a certain proportion with a drug content of 10-500 mg per tablet. Table 8 shows the tablet formulation ratio.

[0113] Table 8. Formulations for preparing cimicifugoside crystal tablets with crystal type II.

[0114]

[0115]

[0116] The method for preparing tablet formulations using pure cimicifugoside crystals of crystal type II as raw materials is as follows: several excipients are mixed evenly with the raw materials, an appropriate amount of 1% sodium hydroxymethyl cellulose solution is added to form a soft material, which is then granulated by sieving, the wet granules are dried, sieved and sized, magnesium stearate and talc are added and mixed evenly, and then tableted to obtain the final product.

[0117] Preparation method 3 of combination drug formulation (capsules)

[0118] Preparation method: Pure cimicifugoside crystals of crystal type II were used as the raw material for the combination drug, and several excipients were used as excipients for preparing the combination drug capsules. Capsule samples with a drug content of 10-500mg per tablet were prepared according to a certain ratio. Table 9 shows the capsule formulation ratio.

[0119] Table 9. Active pharmaceutical ingredient and excipient formulations of cimicifugoside crystal capsules containing crystal type II.

[0120]

[0121] The method for preparing a tablet formulation from a crystalline form of cimicifugoside active pharmaceutical ingredient is as follows: several excipients are mixed evenly with the active pharmaceutical ingredient, an appropriate amount of 1% sodium carboxymethyl cellulose solution is added, wet granules are formed, dried, sieved and granulated, magnesium stearate is added and mixed evenly, and then inserted into capsules; or, without using the granulation step, the crystalline form of cimicifugoside active pharmaceutical ingredient crystals are directly mixed evenly with several excipients and excipients, sieved, and directly filled into capsules.

[0122] Example 4 Dosage Form

[0123] Dosage of cimicifugoside crystals of crystal type II (tablet) 1

[0124] The pharmaceutical composition developed using crystalline type II cimicifugoside crystals as the active pharmaceutical ingredient, with a daily dosage of 0.5-300 mg, can be prepared into ordinary tablets containing 10, 100, 200, 300, and 500 mg of active ingredient, in doses of 1-6 tablets per dose.

[0125] Dosage of type II citric acid crystals 2 (capsules)

[0126] The pharmaceutical composition developed using crystalline type II cimicifugoside crystals as the active pharmaceutical ingredient is characterized by using crystalline type II cimicifugoside crystals as the active pharmaceutical ingredient, with a daily dosage of 10-3000 mg, and can be prepared into capsules of 1-6 capsules per dose, containing 10, 100, 200, 300, and 500 mg of active ingredient.

[0127] References

[0128] [1] Wen Yanli, Huo Shixia, Zhang Wei, et al. Research progress on pharmacological effects and pharmacokinetics of citric acid-like substances [J]. Chinese Journal of Traditional and Folk Medicine, 2015, 24(14): 26-27+29.

[0129] [2] Tan Xue, Gao Li, Ren Jia, et al. Effect of phytoesin on scopolamine-induced learning and memory impairment in mice [J]. Chinese Journal of New Drugs, 2018, 27(23): 2812-2818.

[0130] [3] Pu Jinghua, Pu Xiaoping, Ma Jian, et al. The effect of phytohemagglutinin on scopolamine-induced memory acquisition impairment [J]. Chinese Journal of Pharmacology, 2001, (06): 625-627.

[0131] [4] Ren Jia, Gao Li, Tan Xue, et al. Effects of citric acid-like glycosides on learning and memory abilities and inflammatory factors in rats with cerebral ischemia-reperfusion injury [J]. China Journal of Traditional Chinese Medicine Information, 2019, 26(04): 47-51.

[0132] [5] Ren Jia, Gao Li, Tan Xue, et al. Effects of citric acid on learning and memory function and oxidative stress in rats with vascular dementia [J]. Chinese Journal of Behavioral Medicine and Brain Science, 2018, 27(9): 777-782.

[0133] [6] Chen Jiayuan, Gao Li, Luo Jianan, et al. Effects of citric acid-like substances on behavioral and hormonal changes in APP / PS1 double transgenic mice [J]. China Medical Herald, 2020, 17(08): 9-12.

[0134] [7] Wang Dongyan, Sheng Shudong, Sun Yun. Experimental study on the effect of cimicifugain on D-galactose-induced aging model mice [C] / / Chinese Pharmaceutical Association, China Association of Traditional Chinese Medicine, Hotan Prefectural Committee, Administrative Office. Proceedings of the 6th Symposium on Cistanche deserticola and psammophytic medicinal plants. School of Medicine, Yangzhou Environmental Resources College; School of Medicine, Yangzhou University; 2011: 1.

[0135] [8] Gao Li, Yan Ming, Huo Shixia. Use of phytoescin-like compounds in the preparation of anti-anxiety drugs: CN201611145131.4[P]. CN106581019B.

[0136] [9] Gao Li, Yan Ming, Huo Shixia. Use of phytoesin in the preparation of antidepressant drugs: CN201610390189.9[P].CN105878258B.

Claims

1. A solvate crystal of cimicifugain and n-butanol, wherein the molar ratio of cimicifugain to n-butanol is 1:2, exhibiting crystal type II. When using CuK... α The powder X-ray diffraction pattern of the crystal type II, radiated and expressed in 2θ angles, has the following characteristic peaks: 4.9±0.2°, 7.0±0.2°, 8.2±0.2°, 8.9±0.2°, 11.5±0.2°, 12.6±0.2°, 17.0±0.2°, 19.9±0.2°, 21.0±0.2°, 21.4±0.2°, and 25.4±0.2°.

2. The solvate crystal of cimicifugain and n-butanol as described in claim 1, wherein CuK is used. α Radiation, expressed at a 2θ angle, the powder X-ray diffraction pattern of the type II crystal has the following characteristic peaks and peak area percentages: Preferably, CuK is used. α Radiation, expressed in 2θ angles, and the powder X-ray diffraction pattern of the type II crystal has the following characteristic peaks: 4.9±0.2°, 7.0±0.2°, 8.2±0.2°, 8.9±0.2°, 9.9±0.2°, 11.5±0.2°, 12.6±0.2°, 13.3±0.2°, 14.0±0.2°, 14.4±0.2°, 14.8±0.2°, 15.3±0.2°, 17.0±0.2°, 17.5±0.2°, 17.9±0.2°. 18.6±0.2°, 19.9±0.2°, 20.3±0.2°, 20.8±0.2°, 21.0±0.2°, 21.4±0.2°, 21.6±0.2°, 22.1±0.2°, 22.9±0.2°, 23.2±0.2°, 24.1±0.2°, 24.6±0.2°, 25.4±0.2°, 27.6±0.2°, 29.7±0.2°, 30.0±0.2°, 30.9±0.2°, 45.1±0.2°; Preferably, CuK is used. α Radiation, expressed at a 2θ angle, the powder X-ray diffraction pattern of the type II crystal has the following characteristic peaks and peak area percentages: Preferably, CuK is used. α The radiation is expressed in 2θ angles, and the powder X-ray diffraction pattern of the crystal type II is shown in Figure 1.

3. The solvate crystal of cimicifugain and n-butanol as described in claim 1, wherein when analyzed using attenuated total reflectance Fourier transform infrared spectroscopy, the infrared spectrum of crystal type II is at 3263±2 cm⁻¹. -1 2953±2cm -1 2886±2cm -1 1693±2cm -1 1636±2cm -1 1607±2cm -1 1530±2cm -1 1444±2cm -1 1381±2cm -1 1340±2cm -1 1308±2cm -1 1286±2cm -1 1272±2cm -1 1252±2cm -1 1204±2cm -1 1180±2cm -1 1167±2cm -1 1151±2cm -1 1140±2cm -1 1115±2cm -1 1085±2cm -1 1072±2cm -1 1059±2cm -1 1040±2cm -1 1021±2cm -1 1008±2cm -1 976±2cm -1 906±2cm -1 871±2cm -1 850±2cm -1 829±2cm -1 816±2cm -1 807±2cm -1 784±2cm -1 739±2cm -1 714±2cm -1 There are characteristic infrared spectral peaks at the location; preferably, the infrared spectrum of the crystal type II is shown in Figure 3.

4. The solvate crystal of cimicifugain and n-butanol as described in claim 1, wherein the differential scanning calorimetry (DSC) spectrum of the crystal type II has two endothermic peaks at 106±3℃ and 133±3℃; preferably, the differential scanning calorimetry spectrum of the crystal type II has a heating rate of 10℃ / min in the range of 30 to 200℃.

5. The method for preparing the solvate crystal of cimicifugain and n-butanol according to any one of claims 1-4, wherein the method is a suspension stirring method, a solvent evaporation crystallization method, or a liquid addition grinding method; Preferably, the suspension stirring method includes: Add n-butanol as a single solvent or a mixture of n-butanol and an organic solvent to the cimicifuga glycoside, stir at 300-500 r / min for 72-240 h at a temperature of 20℃-45℃, and dry the product to obtain the crystal with crystal type II. Preferably, the solvent evaporation crystallization method includes: dissolving citric acid in n-butanol as a single solvent or a mixed solvent of n-butanol and water in a volume ratio of 4:1 at a temperature of 15°C to 60°C, wherein the mass of citric acid to the volume ratio of the n-butanol single solvent or the mixed solvent of n-butanol and water is 0.1 mg / mL to 20 mg / mL, stirring for 2 to 24 hours, placing the solution at a temperature of 20°C to 40°C, and slowly evaporating the solvent for 14 to 30 days under a relative humidity of 10% to 90%, until the solvent is completely evaporated, thereby obtaining the crystals having crystal type II. Preferably, the liquid grinding method includes: (1) placing the cimicifuga glycoside in a mortar and grinding it evenly; (2) taking the substance obtained in step (1) and grinding it in a clockwise direction, and adding n-butanol or a mixture of n-butanol and organic solvent dropwise every 15 minutes, with each addition of solvent being 2 mL / g to 10 mL / g, and grinding for 12 to 48 hours; (3) drying the product obtained in step (2) to obtain the crystal with crystal type II. Preferably, the organic solvent is selected from one or more of methanol, ethanol, tetrahydrofuran, acetone, acetonitrile, dioxane, n-hexane, and cyclohexane; Preferably, the volume ratio between any two organic solvents is 1:10 to 10:

1.

6. A solid mixture comprising crystals of a solvate of cimicifugain and n-butanol as described in any one of claims 1-4, wherein the amount of the crystals is 1-99.9% by weight, preferably 10-99.9% by weight, 50-99.9% by weight, or 85-99.9% by weight, based on the weight of the solid mixture.

7. A pharmaceutical composition comprising a therapeutically effective amount of a solvate crystal of any one of claims 1-4 and n-butanol or a solid mixture of claims 6, and a pharmaceutically acceptable carrier; preferably, the daily dose of the pharmaceutical composition is 10-1000 mg; preferably, the pharmaceutical composition is prepared as a tablet, capsule, pill, injectable formulation, powder for injection, granule, powder, micro-pellet, drop, suppository, film, patch, aerosol, or spray; preferably, the pharmaceutical composition is prepared as a sustained-release or controlled-release formulation.

8. Use of the solvate crystals of cimicifugain and n-butanol according to any one of claims 1-4, the solid mixture according to claim 6, or the pharmaceutical composition according to claim 7 in the preparation of a medicament for the prevention and / or treatment of inflammation, anxiety, depression, tumors, and neurodegenerative diseases.

9. Use of the solvate crystals of cimicifugain and n-butanol according to any one of claims 1-4, the solid mixture according to claim 6, or the pharmaceutical composition according to claim 7 in the preparation of a medicament for immunomodulation.

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