Puerarin and nicotinamide eutectic and application thereof

Abstract

The invention discloses an eutectic of puerarin and nicotinamide, the molar ratio of puerarin to nicotinamide is 1: 1, and compared with puerarin, the eutectic has higher solubility, stability, in-vivo absorption speed and onset speed.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular, relates to a cocrystal of puerarin and nicotinamide, a preparation method, a composition and use thereof. Background Art

[0002] Puerarin is an isoflavone derivative with the effect of expanding the crown, which is extracted from the dried root of Pueraria lobata (Willd.) Ohwi, a leguminous plant. Its chemical name is 8-β-D-glucose pyranose-4',7-dihydroxyisoflavone, and its molecular formula is C 21 H 20 O 9 , the structural formula is shown in formula a, which has the effects of preventing and treating cardiovascular diseases, anti-oxidation, anti-cancer, anti-hypertension, hypoglycemic, anti-inflammatory, antipyretic and analgesic, anti-Alzheimer's disease, and anti-osteoporosis (Zhou YX, Zhang H, Peng C. Puerarin: a review of pharmacological effects [J]. Phytotherapy Research, 2014, 28 (7): 961-975).

[0003] The molecular formula of Nicotinamide is C 6 H 6 N 2 O, the structural formula is shown below.

[0004]

[0005] Currently, there is a need to improve the stability, in vivo absorption rate and onset speed of puerarin. Summary of the invention

[0006] One or more embodiments of the present application provide a co-crystal of puerarin and nicotinamide, which has better solubility, stability, in vivo absorption rate and onset rate.

[0007] In one or more embodiments, puerarin and nicotinamide are prepared into a cocrystal solid material with specific non-covalent interaction forces, thereby forming a new substance different from puerarin, nicotinamide and a simple physical mixture of the two. It is further found that the new cocrystal solid material of the present invention has unexpected technical advantages in the preparation of drugs with the effects of preventing and treating cardiovascular diseases, anti-oxidation, anti-cancer, anti-hypertension, hypoglycemic, anti-inflammatory, antipyretic and analgesic, anti-Alzheimer's disease, anti-osteoporosis, etc.

[0008] One or more embodiments of the present application provide a co-crystal of puerarin and nicotinamide, wherein the molar ratio of puerarin to nicotinamide is 1:1.

[0009] In one or more embodiments, CuK α The powder X-ray diffraction pattern of the co-crystal has the following characteristic peaks when irradiated and expressed in 2θ angles: 5.48±0.2°, 8.62±0.2°, 10.14±0.2°, 13.82±0.2°, 15.26±0.2°, 16.20±0.2°, 16.96±0.2°, 17.90±0.2°, and 18.70±0.2°.

[0010] In one or more embodiments, CuK α The powder X-ray diffraction pattern of the co-crystal has the following characteristic peaks and peak area percentages:

[0011] 2θ±0.2° Peak area %±5% 5.48 9.60 8.62 24.40 10.14 9.00 13.82 13.10 15.26 25.00 16.20 100.00 16.96 34.20 17.90 21.30 18.70 92.10 .

[0012] In one or more embodiments, CuK α The powder X-ray diffraction pattern of the co-crystal has the following characteristic peaks: 5.48±0.2°, 8.62±0.2°, 10.14±0.2°, 13.82±0.2°, 15.26±0.2°, 16.20±0.2°, 16.96±0.2°, 17.90±0.2°, 18.70±0.2°, 21.26±0.2°, 21.92±0.2°, 22.46±0.2°, 23.96±0.2°, 25.32±0.2°, 25.88±0.2°, 26.74±0.2°, 27.32±0.2°, 28.72±0.2°, 30.66±0.2°, and 31.50±0.2°.

[0013] In one or more embodiments, CuK α The powder X-ray diffraction pattern of the co-crystal has the following characteristic peaks and peak area percentages:

[0014]

[0015]

[0016] In one or more embodiments, CuK αThe powder X-ray diffraction pattern of the cocrystal has the following characteristic peaks: 5.48±0.2°, 8.62±0.2°, 10.14±0.2°, 13.82±0.2°, 15.26±0.2°, 16.20±0.2°, 16.96±0.2°, 17.90±0.2°, 18.70±0.2°, 21.26±0.2°, 21.92±0.2°, 22.46±0.2°, 23.96±0.2°, 25.32±0.2°, 26.80±0.2°, 27.70±0.2°, 28.60±0.2°, 29.90±0.2°, 30.80±0.2°, 31.30±0.2°, 33.70±0.2°, 34. 0.2°, 25.88±0.2°, 26.74±0.2°, 27.32±0.2°, 28.72±0.2°, 30.66±0.2°, 31.50±0.2°, 32.60±0.2°, 34.84±0.2°, 36.44±0.2°, 38.48±0.2°, 39.66±0.2°, 41.98±0.2°, 42.76±0.2°, 43.60±0.2°, 45.48±0.2°, 46.28±0.2°.

[0017] In one or more embodiments, CuK α The powder X-ray diffraction pattern of the co-crystal has the following characteristic peaks and peak area percentages:

[0018]

[0019]

[0020] In one or more embodiments, CuK α Radiation and expressed in 2θ angles, the powder X-ray diffraction pattern of the co-crystal is as follows Figure 1 shown.

[0021] In one or more embodiments, when analyzed using attenuated total reflection Fourier transform infrared spectroscopy, the infrared spectrum of the co-crystal is at 3485 cm -1 、3342cm -1 、2902cm -1 、1658cm -1 、1628cm -1 、1589cm -1 、1516cm -1 、1446cm -1 、1391cm -1 、1365cm -1 、1303cm -1 、1255cm -1 、1213cm -1 、1179cm -1 、1129cm-1 、1113cm -1 、1068cm -1 、1037cm -1 、1013cm -1 、990cm -1 、889cm -1 、877cm -1 、835cm -1 、824cm -1 、802cm -1 、789cm -1 、742cm -1 、660cm -1 、632cm -1 、568cm -1 、539cm -1 、506cm -1 、488cm -1 、446cm -1 ±2cm -1 There are characteristic peaks in the infrared spectrum.

[0022] In one or more embodiments, the infrared spectrum of the co-crystal is as follows Figure 3 shown.

[0023] In one or more embodiments, in the differential scanning calorimetry spectrum of the co-crystal, there is an endothermic peak at 159.45° C.±3° C.

[0024] In one or more embodiments, within the range of 30-200° C., the heating rate is 10° C. / min.

[0025] One or more embodiments of the present application provide a method for preparing the co-crystal of puerarin and nicotinamide of the present application, which comprises mixing puerarin and nicotinamide in a molar ratio of 1:1, and preparing the co-crystal by an organic solvent liquid grinding method.

[0026] 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.

[0027] In one or more embodiments, the volume ratio between any two of the organic solvents is 1:10 to 10:1 (e.g., 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).

[0028] In one or more embodiments, the amount of liquid added in the organic solvent liquid-adding grinding method is 5 to 100 mL (eg, 50 mL) of organic solvent per gram of the mixed sample of puerarin and nicotinamide.

[0029] In one or more embodiments, the grinding time is 0.1 hour to 10 hours (e.g., 5 hours), the drying temperature is 25-60°C (e.g., 40°C), the drying time is 2 to 10 hours (e.g., 6 hours); the bowl filling rate is 10% to 50% (e.g., 30%), and the reciprocating motion speed is 20 to 70 m / min (meters per minute) (e.g., 45 m / min).

[0030] One or more embodiments of the present application provide a solid mixture, which comprises a co-crystal of puerarin and niacinamide of the present application, wherein the amount of the co-crystal is 1-99.9 wt % (e.g., 10 wt %, 20 wt %, 30 wt %, 40 wt %, 50 wt %, 60 wt %, 70 wt %, 80 wt %, 90 wt %) based on the weight of the solid mixture.

[0031] In one or more embodiments, the amount of the eutectic is 10-99.9 wt %, 50-99.9 wt %, or 85-99.9 wt % based on the weight of the solid mixture.

[0032] One or more embodiments of the present application provide a pharmaceutical composition comprising a therapeutically effective amount of the co-crystal of the present application or the solid mixture of the present application and a pharmaceutically acceptable carrier.

[0033] In one or more embodiments, the daily dosage of the pharmaceutical composition is 10 to 1000 mg (eg, 100 mg, 500 mg, 900 mg).

[0034] In one or more embodiments, the pharmaceutical composition is prepared as a tablet, capsule, pill, or powder injection.

[0035] In one or more embodiments, the pharmaceutical composition is prepared as a sustained-release formulation or a controlled-release formulation.

[0036] One or more embodiments of the present application provide the use of the co-crystal of the present application, the solid mixture of the present application, or the pharmaceutical composition of the present application in the preparation of drugs for preventing and / or treating cardiovascular diseases, anti-oxidation, anti-cancer, hypoglycemic, anti-inflammatory, antipyretic and analgesic, anti-Alzheimer's disease, and anti-osteoporosis.

[0037] In one or more embodiments, in the differential scanning calorimetry spectrum of the co-crystal, there is an endothermic peak at 159.45° C.±3° C.

[0038] In one or more embodiments, at a temperature of 25° C. and a relative humidity range of 0% to 90%, the mass of the eutectic varies with humidity and the adsorption / desorption conditions are as follows: Figure 6 And as shown in Table 4.

[0039] In one or more embodiments, puerarin and nicotinamide are combined with each other by non-covalent bonds to form a co-crystal.

[0040] In one or more embodiments, the molecular formula of the co-crystal formed by puerarin and nicotinamide is C 21 H 20 O 9 ·C 6 H 6 N 2 O.

[0041] In one or more embodiments, puerarin and nicotinamide are prepared into a co-crystal solid material having a specific non-covalent interaction force, thereby forming a new substance different from puerarin, nicotinamide and a physical mixture thereof.

[0042] In one or more embodiments, a mixed solid material or pharmaceutical composition is provided comprising a co-crystal of puerarin and nicotinamide in any non-zero ratio.

[0043] In one or more embodiments, a pharmaceutical composition using a cocrystal of puerarin and nicotinamide as a pharmaceutically active ingredient is provided, wherein the dosage per administration is 10 to 1000 mg. The pharmaceutical composition can be prepared as tablets, capsules, pills, injection preparations, sustained-release or controlled-release preparations.

[0044] In one or more embodiments, the co-crystal of puerarin and nicotinamide of the present application has good solubility.

[0045] Morphological characteristics of puerarin and nicotinamide cocrystal samples

[0046] The co-crystal of puerarin and nicotinamide of the present application is a co-crystal material formed by non-covalent bonding of puerarin and nicotinamide in a molar ratio of 1:1.

[0047] The co-crystal of puerarin and nicotinamide of the present application is analyzed by powder X-ray diffraction using CuK α Under radiation experimental conditions, it is manifested as the diffraction peak position: 2-Theta (°) or Diffraction peak relative intensity: peak height value (Height%) or peak area value (Area%), see Table 1 and Figure 1 .

[0048] The powder X-ray diffraction patterns of the cocrystal of puerarin and nicotinamide and the physical mixture of puerarin and nicotinamide are significantly different in terms of the number of diffraction peaks, the positions of the diffraction peaks, the intensity of the diffraction peaks, the topological patterns of the diffraction peaks, etc. (see Figure 1 and Figure 2 ), indicating that puerarin and niacinamide formed a cocrystal and were different substances from the physical mixture of puerarin and niacinamide.

[0049] Table 1 Powder X-ray diffraction peak data of the cocrystal of puerarin and nicotinamide

[0050]

[0051] Table 2 Powder X-ray diffraction peak data of the physical mixture of puerarin and nicotinamide

[0052]

[0053] The co-crystal of puerarin and nicotinamide of the present application is analyzed by attenuated total reflection Fourier transform infrared spectroscopy at 3485cm -1 、3342cm -1 、2902cm -1 、1658cm -1 、1628cm -1 、1589cm -1 、1516cm -1 、1446cm -1 、1391cm -1 、1365cm -1 、1303cm -1 、1255cm -1 、1213cm -1 、1179cm -1 、1129cm -1 、1113cm -1 、1068cm -1 、1037cm -1 、1013cm -1 、990cm -1 、889cm -1 、877cm -1 、835cm -1 、824cm -1 、802cm -1 、789cm -1 、742cm -1 、660cm -1 、632cm -1 、568cm -1 、539cm -1 、506cm-1 、488cm -1 、446cm -1 There is a characteristic peak of infrared spectrum at the position, and the allowable deviation of the characteristic peak of infrared spectrum is ±2cm- 1 (See Figure 3 ).

[0054] When the cocrystal of puerarin and nicotinamide of the present application is analyzed by differential scanning calorimetry, the DSC spectrum has an exothermic peak at 159.45°C±3°C when the heating rate is 10°C / min in the range of 30-200°C (see Figure 4 ), which is the decomposition peak of the cocrystal. The DSC spectra of the cocrystal of puerarin and nicotinamide are significantly different from those of puerarin and nicotinamide in terms of the number and position of endothermic / exothermic peaks, indicating that puerarin and nicotinamide have formed a new phase, namely the cocrystal of puerarin and nicotinamide (see Figure 5 ).

[0055] Preparation method of puerarin and nicotinamide cocrystal and mixed solid material

[0056] The preparation method of the cocrystal of puerarin and nicotinamide of the present application adopts the liquid addition grinding method, puerarin and nicotinamide are mixed in a molar ratio of 1:1, an organic solvent is added, and the grinding time is 0.1 hour to 10 hours under room temperature, and the drying time is 2 to 10 hours: the filling rate of the bowl of the liquid addition grinding method is 10% to 50%, and the reciprocating speed is 20 to 70 m / min, and the cocrystal of puerarin and nicotinamide is obtained. The organic solvent can be selected from any one or more of methanol, ethanol, acetonitrile, acetone, tetrahydrofuran, dioxane, n-hexane, cyclohexane, and a mixed solvent prepared by combining different proportions.

[0057] The mixed solid material containing the cocrystal of puerarin and nicotinamide of the present application is prepared by mixing the cocrystal of puerarin and nicotinamide obtained by the above method with other chemical substances in any non-zero ratio and in a conventional manner.

[0058] Pharmaceutical preparation composition comprising co-crystal of puerarin and nicotinamide, dosage characteristics and pharmaceutical use The pharmaceutical composition of the present application comprises co-crystal of puerarin and nicotinamide and a pharmaceutically acceptable carrier.

[0059] The pharmaceutical composition of the present application comprises a mixed solid of a co-crystal of puerarin and nicotinamide and a pharmaceutically acceptable carrier.

[0060] The pharmaceutical composition of the present application, comprising the cocrystal of puerarin and nicotinamide, has a daily dosage of 10 to 1000 mg.

[0061] The dosage form of the pharmaceutical composition of the present application can be various tablets, capsules, pills, injection preparations, sustained-release preparations or controlled-release preparations.

[0062] The co-crystal of puerarin and nicotinamide of the present application has the uses of preventing and treating cardiovascular diseases, anti-oxidation, anti-cancer, anti-hypertension, hypoglycemic, anti-inflammatory, antipyretic and analgesic, anti-Alzheimer's disease, and anti-osteoporosis.

[0063] The pharmaceutical composition with the cocrystal of puerarin and nicotinamide as the active ingredient of the present application can be prepared according to methods known in the art. The cocrystal of puerarin and nicotinamide of the present application can be combined with one or more pharmaceutically acceptable solid or liquid excipients and / or adjuvants to prepare any dosage form suitable for human or animal use. The cocrystal of puerarin and nicotinamide of the present application and the mixed solid containing the cocrystal of puerarin and nicotinamide in the pharmaceutical composition are contained in an amount of 10% to 90% by weight.

[0064] The co-crystal of puerarin and nicotinamide of the present application can be administered in a unit dosage form, and the administration route can be enteral or parenteral, such as oral, intravenous, intramuscular, subcutaneous, nasal, oral mucosa, eyes, lungs and respiratory tract, skin, vagina, rectum, etc.

[0065] The dosage form of the present application may be a solid dosage form. The solid dosage form may be tablets (including ordinary tablets, enteric-coated tablets, lozenges, dispersible tablets, chewable tablets, effervescent tablets, orally disintegrating tablets), capsules (including hard capsules, soft capsules, enteric-coated capsules), granules, powders, pellets, dripping pills, suppositories, films, patches, aerosols (powders), sprays, etc.

[0066] The cocrystal of puerarin and nicotinamide and the mixed solid material containing the cocrystal of puerarin and nicotinamide of the present application can be made into ordinary preparations, and can also be made into sustained-release preparations, controlled-release preparations, targeted preparations and various microparticle drug delivery systems.

[0067] The co-crystal of puerarin and nicotinamide of the present application can be formed into tablets, and various excipients known in the art can be widely used, including diluents, binders, wetting agents, disintegrants, lubricants, and glidants. The diluent may be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc.; the wetting agent may be water, ethanol, isopropanol, etc.; the binder may be starch slurry, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, acacia slurry, gelatin slurry, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinyl pyrrolidone, polyethylene glycol, etc.; the disintegrant may be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, cross-linked polyvinyl pyrrolidone, cross-linked sodium carboxymethyl cellulose, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitan fatty acid ester, sodium dodecyl sulfate, etc.; the lubricant and glidant may be talc, silicon dioxide, stearate, tartaric acid, liquid paraffin, polyethylene glycol, etc.

[0068] Tablets can be further made into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double-layer tablets and multi-layer tablets.

[0069] The dosing unit can be made into a capsule, and the cocrystal of the active ingredient puerarin and nicotinamide can be mixed with a diluent and a glidant, and the mixture can be directly placed in a hard capsule or a soft capsule. The cocrystal of the active ingredient puerarin and nicotinamide can also be first made into granules or pellets with a diluent, a binder, and a disintegrant, and then placed in a hard capsule or a soft capsule. Various diluents, binders, wetting agents, disintegrants, and glidants used to prepare the cocrystal tablets of puerarin and nicotinamide can also be used to prepare the capsules of the cocrystal of puerarin and nicotinamide of the present application.

[0070] Furthermore, if necessary, colorants, preservatives, perfumes, flavoring agents or other additives may be added to the pharmaceutical preparations.

[0071] In order to achieve the purpose of medication and enhance the therapeutic effect, the drug of the present invention can be administered by any known administration method.

[0072] The dosage of the pharmaceutical composition of the cocrystal of puerarin and nicotinamide of the present application can vary widely according to the nature and severity of the disease to be prevented or treated, the individual conditions of the patient or animal, the route of administration and the dosage form, etc. The above dosage can be administered in one dosage unit or divided into several dosage units, depending on the clinical experience of the doctor and the dosage regimen including the use of other treatment means.

[0073] The cocrystal or composition of puerarin and nicotinamide of the present application can be taken alone or used in combination with other therapeutic drugs or symptomatic drugs. When the cocrystal of puerarin and nicotinamide of the present invention has a synergistic effect with other therapeutic drugs, its dosage should be adjusted according to the actual situation.

[0074] Beneficial technical effects of the present invention

[0075] The co-crystal of puerarin and nicotinamide of the present application has advantages in terms of safety, solubility, dissolution rate, stability, bioavailability, and in vivo absorption rate.

[0076] The co-crystal of puerarin and nicotinamide of the present application does not contain any crystallization solvent and has the advantage of good safety in drug formulation.

[0077] The cocrystal of puerarin and nicotinamide of the present application is superior to the puerarin raw material in terms of solubility, and exhibits a dissolution rate substantially consistent with that of the puerarin raw material in pure water ( Figure 6 ).

[0078] The cocrystal of puerarin and nicotinamide of the present application can be stably present under high temperature (60°C), high humidity (25°C, relative humidity 90%±5%), and light (4500lx±500lx) conditions, which is different from the puerarin raw material which is unstable under high temperature, high humidity and light conditions ( Figure 7 ) has a better stability advantage ( Figure 8 ).

[0079] The co-crystal of puerarin and nicotinamide of the present application is used as the active ingredient of the drug and the pharmaceutical composition thereof. After oral administration, the drug is rapidly and massively released through the gastrointestinal tract or blood. The peak time of puerarin (T max ) was shortened by 0.5 hours, and the half-life (t 1 / 2 ) has also been shortened, playing an advantageous role in preventing and treating diseases ( Fig. 9 ).

[0080] There are many factors that affect the dosage of the active ingredients of the pharmaceutical composition of the cocrystal of puerarin and nicotinamide of the present application, such as: different daily dosages due to different uses for prevention and treatment; different daily dosages due to different natures and severity of illness; different daily dosages due to different genders, ages, and body surface areas of patients, different routes of administration, number of administrations, and different treatment purposes. The daily dosage range of the cocrystal of puerarin and nicotinamide used in the present invention is 0.002-20 mg / kg body weight, such as 0.01-1 omg / kg body weight. When used, different total dosage plans of the active ingredients of the cocrystal of puerarin and nicotinamide should be formulated according to the actual prevention and treatment requirements of different situations, and can be completed in multiple or one-time administration. BRIEF DESCRIPTION OF THE DRAWINGS

[0081] Figure 1 This is the powder X-ray diffraction pattern of the co-crystal of puerarin and niacinamide.

[0082] Figure 2 It is the powder X-ray diffraction pattern of the physical mixture of puerarin and niacinamide.

[0083] Figure 3 This is the infrared absorption spectrum of the co-crystal of puerarin and niacinamide.

[0084] Figure 4 This is the differential scanning calorimetry spectrum of the co-crystal of puerarin and nicotinamide.

[0085] Figure 5 It is the differential scanning calorimetry spectrum of the cocrystal of puerarin and niacinamide and the raw materials of puerarin and niacinamide.

[0086] Figure 6 This is the solubility curve of the cocrystal of puerarin and nicotinamide in pure water (pH 7.0).

[0087] Figure 7 This is the stability spectrum of puerarin raw material.

[0088] Figure 8 This is the stability spectrum of the co-crystal of puerarin and niacinamide.

[0089] Fig. 9 This is the blood concentration-time curve of puerarin in the cocrystal of puerarin and niacinamide (0-12h). DETAILED DESCRIPTION

[0090] In order to better illustrate the technical solution of the present invention, the following embodiments are given, but the present invention is not limited thereto.

[0091] Example 1

[0092] Preparation of co-crystal of puerarin and nicotinamide

[0093] Take appropriate amount of puerarin and nicotinamide, the molar ratio of the two is 1:1, add the sample to a mortar of appropriate capacity at room temperature by liquid grinding method, add appropriate amount of organic solvent, grind at room temperature at appropriate speed for appropriate time, and dry the obtained powder or paste sample naturally or under reduced pressure. The condition parameters of the preparation method are shown in Table 3. Powder X-ray diffraction analysis is performed on it, and the diffraction patterns are consistent with Figure 1 The results are consistent, indicating that the obtained sample is a co-crystal of puerarin and nicotinamide.

[0094] Table 3 Specific examples of the preparation method of the co-crystal of puerarin and nicotinamide

[0095]

[0096] Example 2

[0097] In vitro solubility characteristics of cocrystal of puerarin and nicotinamide

[0098] The solubility characteristics of the cocrystal of puerarin and nicotinamide and puerarin raw materials in water were investigated. The experiment was conducted in accordance with the Technical Guidelines for Dissolution Test of Ordinary Oral Solid Preparations. The puerarin content was determined at a wavelength of 250nm by high performance liquid chromatography, and its dissolved amount was calculated by the external standard method. Dissolution curves were drawn with time as the horizontal axis and the dissolved amount as the vertical axis ( Figure 6 ). The specific data are shown in Table 4:

[0099] Table 4 Dissolution curve data

[0100]

[0101] From the solubility data, it can be seen that the dissolution behavior of the cocrystal of puerarin and nicotinamide in pure water is better than that of the raw material of puerarin. The dissolution amount of the cocrystal of puerarin and nicotinamide is higher than that of the raw material of puerarin. The cocrystal of puerarin and nicotinamide shows basically the same dissolution rate as the raw material of puerarin, and the solubility curve shows a stable release platform.

[0102] Example 3

[0103] Stability of cocrystal of puerarin and nicotinamide

[0104] High temperature test: The co-crystal sample of puerarin and nicotinamide was placed in an open clean surface dish at 60°C for 10 days, and samples were taken on the 0th, 5th and 10th days. The samples obtained from the above sampling points were subjected to powder X-ray diffraction analysis, and the results showed that the co-crystal of puerarin and nicotinamide was stable under the high temperature influence factor test.

[0105] High humidity test: The cocrystal sample of puerarin and nicotinamide was placed in an open clean surface dish at 25°C and relative humidity of 90%±5% for 10 days, and samples were taken on the 0th day, the 5th day and the 10th day. The samples obtained from the above sampling points were subjected to powder X-ray diffraction analysis, and the results showed that the cocrystal of puerarin and nicotinamide was stable under high humidity conditions.

[0106] Light test: The co-crystal sample of puerarin and nicotinamide was placed in an open clean surface dish, placed in a light box equipped with a fluorescent lamp, and placed at an illumination of 4500lx±500lx for 10 days, and samples were taken on the 0th day, the 5th day, and the 10th day. The samples obtained from the above sampling points were subjected to powder X-ray diffraction analysis, and the results showed that the co-crystal of puerarin and nicotinamide was stable under light conditions.

[0107] The stability study of the cocrystal of puerarin and nicotinamide showed that the cocrystal remained stable for 10 days in a high humidity and light environment (see Figure 8 ), which improves the defect of puerarin raw material being unstable in high temperature, high humidity and light environment (see Figure 7 ), and has good drugability characteristics.

[0108] Example 4

[0109] Biological advantages of the cocrystal of puerarin and nicotinamide

[0110] Standard solution: accurately weigh 5.00 mg of puerarin standard, add 50 mL of methanol to fully dissolve and mix, prepare 100 μg / mL puerarin stock solution, store at -20°C, and dilute with methanol to make standard solutions of different concentrations before use. Internal standard solution: accurately weigh 5.00 mg of chrysin, add 50 mL of methanol to fully dissolve and mix, prepare 100 μg / mL chrysin stock solution, store at -20°C, and dilute with methanol to make 100 ng / mL internal standard working solution before use.

[0111] SD rats were randomly divided into 2 groups, 6 in each group, and raised under conventional feeding conditions with free access to water. After fasting for 12 hours, puerarin and the cocrystal of puerarin and nicotinamide were respectively given by solid gavage at a dose of 100 mg / kg. At 5 minutes, 15 minutes, 30 minutes, 1 hour, 1.5 hours, 2 hours, 4 hours, 6 hours, 8 hours, and 12 hours after administration, about 0.3 mL of blood was collected from the posterior ocular venous plexus into a heparinized anticoagulation tube, centrifuged at 4°C and 5000 rpm for 10 minutes, and 100 μL of the supernatant was frozen in a -80°C refrigerator for testing.

[0112] During the determination, 100 μL of plasma was precisely drawn, 10 μL of internal standard chrysin working solution was added, and then 300 μL of acetonitrile was added, fully shaken for 3 min, centrifuged (13000 rpm, 10 min), 380 μL of the upper solution was blown with nitrogen until dry, and then 100 μL of 50% methanol was added to re-dissolve, fully shaken for 3 min, centrifuged (13000 rpm, 3 min), 90 μL of the upper solution was taken into the inner liner tube, and 3 μL was injected. The standard curve was detected by the same method, and the blood concentration of puerarin and nicotinamide was calculated by substituting the ratio of the peak area of ​​the drug to the internal standard into the standard curve, and the blood concentration-time curve was drawn.

[0113] Detection conditions: Chromatographic column: ACQUITY BEH C 18 (2.1×50mm, 1.7μμm); mobile phase A is 0.1% formic acid, mobile phase B is acetonitrile. Column temperature is 40°C; injection volume: 3μL; running time: 7min. Gradient elution conditions are shown in Table 5:

[0114] Table 5 Liquid phase detection gradient elution conditions

[0115]

[0116] Mass spectrometry conditions: negative ion mode, mass-to-charge ratio of Pueraria root is 415.2, and mass-to-charge ratio of Populus is 253.1.

[0117] The pharmacokinetic parameters of puerarin blood concentration data of puerarin and puerarin-nicotinamide cocrystal were calculated. The main pharmacokinetic parameter results are shown in Table 6. The 0-24 hour blood concentration-time change curve is shown in Fig. 9 shown.

[0118] Table 6 Main pharmacokinetic parameters of puerarin in puerarin API and cocrystal of puerarin and nicotinamide

[0119]

[0120] (mean ± standard error)

[0121] From the above experimental research data, it can be seen that the puerarin and nicotinamide cocrystal has a similar blood concentration and bioavailability to puerarin, that is, an effective blood concentration, and a longer peak time (T max ) was shortened by 0.5 hours, and the half-life (t 1 / 2 ) is also shortened, allowing it to reach a higher therapeutic concentration faster than puerarin raw material, making it more suitable for diseases that require rapid release and faster onset of action, such as ischemic angina.

[0122] Example 5

[0123] Preparation method 1 of co-crystal pharmaceutical preparation (tablets):

[0124] Preparation method: The pure cocrystal of puerarin and nicotinamide is used as the raw material of the combination drug, and several excipients are used as the auxiliary material components of the tablets. Tablet samples with a drug content of 10 to 500 mg per tablet are prepared according to a certain ratio. Table 7 shows the tablet formula ratio:

[0125] Table 7 Preparation formula of puerarin and nicotinamide cocrystal tablets

[0126]

[0127] The method of preparing a tablet preparation using the pure cocrystal of puerarin and nicotinamide as a raw material is as follows: uniformly mixing several excipients with the raw material and directly tableting; or mixing the excipients and dry granulating them, and then uniformly mixing them with the raw material and tableting them.

[0128] Preparation method 2 of combined pharmaceutical preparation (tablets):

[0129] Preparation method: The pure cocrystal of puerarin and nicotinamide is used as the raw material, and several excipients are used as the auxiliary ingredients of the tablets. Tablet samples with a drug content of 10 to 500 mg per tablet are prepared in a certain ratio. Table 8 gives the tablet formula ratio.

[0130] Table 8 Preparation formula of puerarin and nicotinamide cocrystal tablets

[0131]

[0132] The method for preparing a tablet preparation using the pure cocrystal of puerarin and nicotinamide as a raw material is as follows: uniformly mixing several excipients with the raw material, adding an appropriate amount of 1% sodium hydroxymethylcellulose solution to prepare a soft material, sieving granules, drying the wet granules, sieving whole granules, adding magnesium stearate and talcum powder, mixing uniformly, and tableting to obtain the tablet preparation.

[0133] Preparation method 3 of combined pharmaceutical preparation (capsules):

[0134] Preparation method: Use the pure cocrystal of puerarin and nicotinamide as the raw material of the combination drug, use several excipients as auxiliary ingredients for preparing the combination drug capsules, and prepare capsule samples with a drug content of 10 to 500 mg per tablet according to a certain ratio. Table 9 gives the capsule formula ratio.

[0135] Table 9 API and excipient formula of co-crystal capsule preparation of puerarin and nicotinamide

[0136]

[0137] The method for preparing a tablet preparation from a co-crystal drug substance of puerarin and nicotinamide is as follows: uniformly mixing several excipients with the drug substance, adding an appropriate amount of 1% sodium hydroxymethylcellulose solution to prepare wet granules, drying and sieving the whole granules, adding magnesium stearate and mixing uniformly, and inserting the granules into capsules; or without using a granulation step, directly mixing the co-crystal drug substance of puerarin and nicotinamide with several excipients and auxiliary materials, sieving, and directly filling the tablet preparation into capsules.

[0138] Example 6

[0139] Dosage of co-crystal of puerarin and nicotinamide 1 (tablet):

[0140] The pharmaceutical composition developed by using the co-crystal sample of puerarin and nicotinamide as the active ingredient of the drug, the co-crystal of puerarin and nicotinamide is used as the active ingredient of the drug, the daily dosage is 0.5-300 mg, and can be prepared into ordinary tablet types of 1-6 tablets each time, containing 10, 100, 200, 300, and 500 mg of the active ingredient.

[0141] Dosage of co-crystal of puerarin and nicotinamide 2 (capsule):

[0142] A pharmaceutical composition is developed using a sample of a cocrystal of puerarin and nicotinamide as a pharmaceutical active ingredient. The cocrystal of puerarin and nicotinamide is used as a pharmaceutical active ingredient, and the daily dosage is 10-3000 mg. The cocrystal can be prepared into capsules containing 10, 100, 200, 300, and 500 mg of the active ingredient, 1 to 6 capsules each time.

Claims

1. A cocrystal of puerarin and nicotinamide, wherein the molar ratio of puerarin to nicotinamide is 1:

1.

2. The eutectic of claim 1, wherein CuK α Radiation and expressed in 2θ angles, the powder X-ray diffraction pattern of the cocrystal has the following characteristic peaks: 5.48±0.2°, 8.62±0.2°, 10.14±0.2°, 13.82±0.2°, 15.26±0.2°, 16.20±0.2°, 16.96±0.2°, 17.90±0.2°, 18.70±0.2°; Preferably, CuK is used α The powder X-ray diffraction pattern of the co-crystal has the following characteristic peaks and peak area percentages: ； Preferably, CuK is used α The powder X-ray diffraction pattern of the co-crystal has the following characteristic peaks: 5.48±0.2°, 8.62±0.2°, 10.14±0.2°, 13.82±0.2°, 15.26±0.2°, 16.20±0.2°, 16.96±0.2°, 17.90±0.2°, 18.70±0.2°, 21.26±0.2°, 21.92±0.2°, 22.46±0.2°, 23.96±0.2°, 25.32±0.2°, 25.88±0.2°, 26.74±0.2°, 27.32±0.2°, 28.72±0.2°, 30.66±0.2°, 31.50±0.2°; Preferably, CuK is used α The powder X-ray diffraction pattern of the co-crystal has the following characteristic peaks and peak area percentages: Preferably, CuK is used α The powder X-ray diffraction pattern of the cocrystal has the following characteristic peaks: 5.48±0.2°, 8.62±0.2°, 10.14±0.2°, 13.82±0.2°, 15.26±0.2°, 16.20±0.2°, 16.96±0.2°, 17.90±0.2°, 18.70±0.2°, 21.26±0.2°, 21.92±0.2°, 22.46±0.2°, 23.96±0.2°, 25.32±0.2°, 26.80±0.2°, 27.70±0.2°, 28.60±0.2°, 29.90±0.2°, 30.80±0.2°, 31.30±0.2°, 33.70±0.2°, 34. 0.2°, 25.88±0.2°, 26.74±0.2°, 27.32±0.2°, 28.72±0.2°, 30.66±0.2°, 31.50±0.2°, 32.60±0.2°, 34.84±0.2°, 36.44±0.2°, 38.48±0.2°, 39.66±0.2°, 41.98±0.2°, 42.76±0.2°, 43.60±0.2°, 45.48±0.2°, 46.28±0.2°; Preferably, CuK is used α The powder X-ray diffraction pattern of the co-crystal has the following characteristic peaks and peak area percentages: Preferably, using CuKα radiation and expressed in 2θ angles, the powder X-ray diffraction pattern of the co-crystal is as shown in FIG1 .

3. The eutectic material according to claim 1, wherein when analyzed by attenuated total reflection Fourier transform infrared spectroscopy, the infrared spectrum of the eutectic material is at 3485 cm -1 、3342cm -1 、2902cm -1 、1658cm -1 、1628cm -1 、1589cm -1 、1516cm -1 、1446cm -1 、1391cm -1 、1365cm -1 、1303cm -1 、1255cm -1 、1213cm -1 、1179cm -1 、1129cm -1 、1113cm -1 、1068cm -1 、1037cm -1 、1013cm -1 、990cm -1 、889cm -1 、877cm -1 、835cm -1 、824cm -1 、802cm -1 、789cm -1 、742cm -1 、660cm -1 、632cm -1 、568cm -1 、539cm -1 、506cm -1 、488cm -1 、446cm -1 ±2cm -1 There is a characteristic peak of infrared spectrum at; preferably, the infrared spectrum of the eutectic is as shown in Figure 3.

4. The eutectic according to claim 1, wherein in the differential scanning calorimetry spectrum of the eutectic, there is an endothermic peak at 159.45°C±3°C; preferably, in the range of 30 to 200°C, the heating rate is 10°C / min.

5. The method for preparing the cocrystal of puerarin and nicotinamide according to any one of claims 1 to 4, comprising mixing puerarin and nicotinamide in a molar ratio of 1:1, and preparing the cocrystal by an organic solvent liquid grinding method; 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 of the organic solvents is 1:10 to 10:1; Preferably, the amount of liquid added in the organic solvent liquid grinding method is 5 to 100 mL of organic solvent per gram of the mixed sample of puerarin and nicotinamide; the grinding time is 0.1 hour to 10 hours, the drying temperature is 25-60° C., and the drying time is 2 to 10 hours; the bowl filling rate is 10% to 50%, and the reciprocating motion speed is 20 to 70 m / min.

6. A solid mixture comprising the co-crystal of puerarin and nicotinamide according to any one of claims 1 to 4, wherein the amount of the co-crystal is 1-99.9 wt %, preferably 10-99.9 wt %, more preferably 50-99.9 wt %, and most preferably 85-99.9 wt %, based on the weight of the solid mixture.

7. A pharmaceutical composition comprising a therapeutically effective amount of the cocrystal according to any one of claims 1 to 4 or the solid mixture according to claim 6 and a pharmaceutically acceptable carrier; preferably, the single-day dosage of the pharmaceutical composition is 10 to 1000 mg; preferably, the pharmaceutical composition is prepared as a tablet, capsule, pill, or powder injection; preferably, the pharmaceutical composition is prepared as a sustained-release preparation or a controlled-release preparation.

8. Use of the cocrystal according to any one of claims 1 to 4, the solid mixture according to claim 6, or the pharmaceutical composition according to claim 7 in the preparation of drugs for preventing and / or treating cardiovascular diseases, anti-oxidation, anti-cancer, hypoglycemic, anti-inflammatory, antipyretic and analgesic, anti-Alzheimer's disease, and anti-osteoporosis.

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