Eutectic of puerarin and berberine hydrochloride, preparation method, composition and application thereof
By preparing a 1:1 molar ratio cocrystal of puerarin and berberine hydrochloride, the problem of instability when administered alone was solved, solubility and bioavailability were improved, the frequency of administration was simplified, and the therapeutic effect was enhanced.
Patent Information
- Application Number
- CN202511765596.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-11-27
- Publication Date
- 2026-02-27
AI Technical Summary
Puerarin and berberine hydrochloride are unstable when administered alone, and are prone to hygroscopic transformation and crystallization. Combined administration increases the frequency of medication for patients, affecting treatment compliance. Furthermore, compound preparations have differences in solubility and stability issues.
A 1:1 molar ratio eutectic of puerarin and berberine hydrochloride was prepared and characterized by powder X-ray diffraction, infrared spectroscopy and differential scanning calorimetry. The eutectic was prepared by grinding or dissolving to form an orthorhombic symmetric eutectic.
It improves the stability and solubility of the cocrystal, enhances the bioavailability of the drug, simplifies the frequency of administration, and improves the therapeutic effect and patient compliance.
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Abstract
Description
TECHNICAL FIELD
[0001] The present application belongs to the technical field of medicine. Specifically, it relates to a co-crystal of puerarin and berberine hydrochloride, a preparation method, a composition and a use thereof. BACKGROUND
[0002] Gegenhuanglian Decoction is a famous Chinese medicine formula, which is recorded in Volume 54 of Jiaoyi Leiju, Tongzhenzi Shanghan Kuaiyao, Jiaofang Jiyijie, Shanghan Da Bai, Chinese Medical Dictionary, etc. It has the effects of relieving the exterior and interior, clearing heat and stopping diarrhea. Modern pharmacology shows that it has the effects of anti-arrhythmia, anti-inflammatory and hypothermia. In the formula, the main component of Gegen is puerarin, and the main component of Huanglian is berberine.
[0003] A drug co-crystal refers to a process of forming a new chemical entity between a drug molecule and a co-crystal former (co-crystal coformer) through non-covalent interactions such as hydrogen bonds and van der Waals forces, which is an important way for new drug research and development. Traditional co-crystal formers are generally inorganic acids, bases, etc. The co-crystal formation can improve the physical and chemical properties such as stability and solubility, so as to improve the drug property. Generally, these introduced chemical substances do not have pharmacological activity. In recent years, drug-drug co-crystals have provided a new idea for drug research and development. Generally, a compound with synergistic effect or activity related to the modified drug is selected to form a co-crystal to form a new chemical entity, which not only can improve the physical and chemical properties of the compound and improve the drug property, but also can reduce the frequency of drug administration and increase patient compliance in clinical practice, which greatly improves the innovation and market competitiveness of the drug.
[0004] Puerarin (PUE) is a natural isoflavone carbon glycoside, which is often used in clinical practice in the form of anhydrous or monohydrate solid preparation (mainly tablets or capsules), and is a drug for treating cardiovascular and cerebrovascular diseases. Puerarin is a white needle-like crystal, which is soluble in hot water. Recent studies have shown that puerarin can remove free radicals in the body to varying degrees, reduce lipid peroxidation damage, and has the effects of improving immunity, improving microcirculation and dilating blood vessels. At the same time, puerarin can also reduce blood sugar to achieve the purpose of treating diabetes.
[0005] Berberine (BER) is an isoquinoline alkaloid, which is often used in clinical practice in the form of hydrochloride solid preparation (mainly tablets or capsules), and is a first-line drug for treating gastrointestinal inflammation, bacterial dysentery and other diseases. Berberine hydrochloride is a yellow crystalline powder, which is soluble in hot water and slightly soluble in cold water or ethanol. Recent studies have shown that berberine hydrochloride also has effects on pulmonary tuberculosis, scarlet fever, acute tonsillitis and respiratory tract infections.
[0006] The chemical name of puerarin is 4,7-dihydroxy-8- β -D-glucosyl isoflavone, and the molecular formula is C21 H 20 O9, as shown in a. The chemical name of berberine hydrochloride is 5,6-dihydro-9,10-dimethoxybenzo[g]-1,3-benzodioxolane[5,6-a]quinolizine hydrochloride dihydrate, and the molecular formula is C 20 H 18 NO4.Cl, as shown in b.
[0007]
[0008] The anhydrate crystal of puerarin and the hydrate crystal of berberine are both unstable, and will absorb moisture and change crystal at room temperature. In addition, due to the complex mechanism of inflammation, cardiovascular disease or diabetes, it is difficult to achieve the expected therapeutic effect by single drug administration, and therefore the combination administration is often used in clinic, which increases the frequency of drug administration and reduces the compliance of patients to disease treatment, and is not conducive to the treatment and management of the disease. At present, researchers have prepared compound preparations according to the clinical prescription amount or traditional Chinese medicine compound, and it is worth noting that the compound preparations may have large solubility difference and poor stability, which greatly limits their clinical application, which brings new challenges to researchers. SUMMARY
[0009] One or more embodiments of the present application provide a co-crystal of puerarin and berberine hydrochloride, wherein the molar ratio of puerarin to berberine hydrochloride is 1:1, and the co-crystal is prepared by using Cu K α Radiation and expressed in 2θ angle, the powder X-ray diffraction pattern of the co-crystal has the following characteristic peaks: 10.2±0.2°, 11.3±0.2°, 12.4±0.2°, 12.8±0.2°, 13.6±0.2°, 14.0±0.2°, 15.1±0.2°, 15.5±0.2°, 15.9±0.2°, 16.2±0.2°, 17.0±0.2°, 17.3±0.2°, 18.2±0.2°, 19.12±0.2°, 19.8±0.2°.
[0010] In one or more embodiments, the Cu K α Radiation and expressed in 2θ angle, the powder X-ray diffraction pattern of the co-crystal has the following characteristic peaks and peak area percentage:
[0012] In one or more embodiments, the Cu K αThe powder X-ray diffraction pattern of the co-crystal has the following characteristic peaks, expressed in degrees 2-theta and measured using Cu radiation:
[0013] Preferably, Cu radiation is used K α The powder X-ray diffraction pattern of the co-crystal has the following characteristic peaks, expressed in degrees 2-theta and measured using Cu radiation:
[0015] In one or more embodiments, Cu radiation is used K α The powder X-ray diffraction pattern of the co-crystal is as shown in Figure 3
[0016] In one or more embodiments, the infrared spectrum of the co-crystal, when analyzed using Fourier transform infrared spectroscopy, has peaks at 3558±2 cm -1 , 3345±2 cm -1 , 3240±2 cm -1 , 3132±2 cm -1 , 2946±2 cm -1 , 2899±2 cm -1 , 2838±2 cm -1 , 1621±2 cm -1 , 1590±2 cm -1 , 1509±2 cm -1 , 1486±2 cm -1 , 1439±2 cm -1 , 1389±2 cm -1 , 1362±2 cm -1 , 1339±2 cm-1 , 1300 ± 2 cm -1 , 1273 ± 2 cm -1 , 1254 ± 2 cm -1 , 1234 ± 2 cm -1 , 1215 ± 2 cm -1 , 1172 ± 2 cm -1 , 1141 ± 2 cm -1 , 1098 ± 2 cm -1 , 1060 ± 2 cm -1 , 1037 ± 2 cm -1 , 1002 ± 2 cm -1 , 936 ± 2 cm -1 , 913 ± 2 cm -1 , 885 ± 2 cm -1 , 831 ± 2 cm -1 , 792 ± 2 cm -1 , 750 ± 2 cm -1 , 707 ± 2 cm -1 , 626 ± 2 cm -1 , 537 ± 2 cm -1 , 506 ± 2 cm -1 , 416 cm -1 .
[0017] In one or more embodiments, the infrared spectrum of the co-crystal is as shown in Figure 5 .
[0018] In one or more embodiments, when analyzed using single crystal X-ray diffraction, the co-crystal exhibits orthorhombic symmetry, space group P 212121, with cell parameter values of a = 7.65528 , b = 15.07721, c = 30.1929, a = 90.00°, b = 90.00°, g = 90.00°, V = 3484.87, Z = 4.
[0019] In one or more embodiments, the differential scanning calorimetry profile of the co-crystal comprises one major endothermic peak at 251 ± 5°C and one minor endothermic peak at 258 ± 5°C when the rate of temperature increase is 10 °C per minute.
[0020] In one or more embodiments, the differential scanning calorimetry profile of the co-crystal is as shown in Figure 6 .
[0021] One or more embodiments of the present application provide a method of preparing a co-crystal, comprising the following method: (1) dissolving puerarin and berberine hydrochloride in an organic solvent at a molar ratio of 1:0.9-1:1.1 (e.g. 1:1) at room temperature to obtain the co-crystal; for example, completely dissolving at 50 °C water bath, standing at room temperature for 7 days; or (2) grinding puerarin and berberine hydrochloride in a mortar at a molar ratio of 1:0.9-1:1.1 (e.g. 1:1), then adding an organic solvent for grinding, and drying to obtain the co-crystal; or (3) dissolving puerarin and berberine hydrochloride in an organic solvent at a molar ratio of 1:0.9-1:1.1 (e.g. 1:1), filtering, and evaporating to obtain the co-crystal.
[0022] In one or more embodiments, the method (2) comprises the following steps: 1) placing puerarin and berberine hydrochloride in a mortar at a molar ratio of 1:0.9-1:1.1 (e.g. 1:1), grinding uniformly to obtain a mixture; 2) grinding the mixture obtained in step 1) (e.g. grinding in a clockwise direction), and adding an organic solvent dropwise thereto every 15 min, grinding for 2 h to obtain a product; 3) drying the product obtained in step 2) to obtain a solid product, the co-crystal of puerarin and berberine hydrochloride.
[0023] In one or more embodiments, the organic solvent is selected from one or more of ethanol, acetonitrile, acetone, isopropanol, and methanol.
[0024] In one or more embodiments, the volume ratio between any two of the organic solvents is 1:10-10:1 (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).
[0025] One or more embodiments of the present application provide a solid mixture comprising the co-crystal of the present application, the amount of the co-crystal being 1-99.9% by weight (e.g. 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% by weight) based on the weight of the solid mixture.
[0026] In one or more embodiments, the amount of the co-crystal is 10-99.9%, 50-99.9%, 85-99.9% by weight based on the weight of the solid mixture.
[0027] One or more embodiments of the present application provide a pharmaceutical composition comprising a prophylactically and / or therapeutically effective amount of the co-crystal of the present application or the solid mixture of the present application and a pharmaceutically acceptable carrier.
[0028] In one or more embodiments, the pharmaceutical composition is administered in a daily dose of 5-3000 mg (e.g., 100 mg, 1000 mg, 2000 mg).
[0029] In one or more embodiments, the pharmaceutical composition is prepared as a tablet, a capsule, a pill, an injection, a granule, a powder, a suppository, a film, a patch, an aerosol or a spray.
[0030] In one or more embodiments, the pill is a pellet or a drop pill.
[0031] In one or more embodiments, the pharmaceutical composition is prepared as a sustained release preparation or a controlled release preparation.
[0032] One or more embodiments of the present application provide 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 a medicament for preventing and / or treating an inflammatory disease, a cardiovascular disease, diabetes, bacterial dysentery, tuberculosis, scarlet fever, acute tonsillitis or a respiratory tract infection.
[0033] In one or more embodiments, the inflammatory disease is gastrointestinal inflammation.
[0034] One or more embodiments of the present application provide a co-crystalline material state and a characterization method of puerarin and berberine hydrochloride.
[0035] One or more embodiments of the present application provide a preparation method of a new, crystalline and stable co-crystal of puerarin and berberine hydrochloride.
[0036] One or more embodiments of the present application provide a pure co-crystal of puerarin and berberine hydrochloride or a mixed solid material containing any non-zero proportion of the co-crystal of puerarin and berberine hydrochloride and a pharmaceutical composition thereof.
[0037] One or more embodiments of the present application provide a pharmaceutical composition using the co-crystal of puerarin and berberine hydrochloride as a pharmaceutically active ingredient, the daily dose of the co-crystal of puerarin and berberine hydrochloride being in the range of 5-3000 mg. The pharmaceutical composition includes a tablet, a capsule, a pill, an injection preparation, a granule, a powder, a pellet, a drop pill, a suppository, a film, a patch, an aerosol, a spray, a sustained release preparation or a controlled release preparation.
[0038] One or more embodiments of the present application provide a co-crystal of puerarin and berberine hydrochloride, which exhibits better solubility advantage in water compared with puerarin or berberine hydrochloride.
[0039] One or more embodiments of the present application provide an application of the co-crystal of puerarin and berberine hydrochloride as a pharmaceutical effective component in the preparation of anti-inflammatory, cardiovascular system and anti-diabetic drugs.
[0040] To solve the above technical problems, the present application adopts the following technical solutions: 1. Morphological characteristics of the co-crystal of puerarin and berberine hydrochloride: 1.1 The co-crystal of puerarin and berberine hydrochloride according to the present application is formed by puerarin and berberine hydrochloride in a 1:1 molar ratio.
[0041] 1.2 The co-crystal of puerarin and berberine hydrochloride according to the present application exhibits orthorhombic symmetry when analyzed by single crystal X-ray diffraction, and the space group is P 212121, the cell parameter values are a=7.65528 , b=15.07721, c=30.1929, α=90.00°, β=90.00°, γ=90.00°, V=3484.87, Z=4 (see Figure 1 , Figure 2 ).
[0042] 1.3 The co-crystal of puerarin and berberine hydrochloride according to the present application, when analyzed by powder X-ray diffraction under the following radiation experimental conditions, Cu K α , the diffraction peak position: 2-Theta value (°) or d value (Å), diffraction peak relative intensity: peak height value (Height%) or peak area value (Area%) has the following characteristics (Table 1, Figure 3 ). The physical mixture powder X-ray diffraction pattern and data of puerarin and berberine hydrochloride are shown in Table 2, Figure 4 The co-crystal of puerarin and berberine hydrochloride and the physical mixture of puerarin and berberine hydrochloride have obvious differences in the number of diffraction peaks, diffraction peak position, diffraction peak intensity, diffraction peak topological pattern, etc., indicating that the co-crystal of puerarin and berberine hydrochloride is not the same as the physical mixture of puerarin and berberine hydrochloride.
[0043] Table 1 Powder X-ray diffraction peak value of the co-crystal of puerarin and berberine hydrochloride
[0044] Table 2 Powder X-ray diffraction peak value of the physical mixture of puerarin and berberine hydrochloride
[0045] 1.4 The co-crystal of puerarin and berberine hydrochloride involved in the present application has infrared spectral characteristic peaks at 3558, 3345, 3240, 3132, 2946, 2899, 2838, 1621, 1590, 1509, 1486, 1439, 1389, 1362, 1339, 1300, 1273, 1254, 1234, 1215, 1172, 1141, 1098, 1060, 1037, 1002, 936, 913, 885, 831, 792, 750, 707, 626, 537, 506, 416 cm -1 -2 cm -1 . Figure 5 .
[0046] 1.5 The co-crystal of puerarin and berberine hydrochloride involved in the present application has a main endothermic peak at 251 ± 5°C and a secondary endothermic peak at 258 ± 5°C in a DSC spectrum when the temperature rising rate is 10°C per minute Figure 6 . The DSC and TG superimposed spectrum of puerarin, berberine hydrochloride and the co-crystal of puerarin and berberine hydrochloride is shown in Figure 7 , Figure 8 . The DSC spectra of puerarin, berberine hydrochloride and the co-crystal of puerarin and berberine hydrochloride are obviously different in the number and position of endothermic / exothermic peaks, indicating that the co-crystal of puerarin and berberine hydrochloride is formed in the form of co-crystal to form a new substance.
[0047] 2. The preparation method of the co-crystal of puerarin and berberine hydrochloride and the mixed solid substance is characterized in that: 2.1 The preparation method of the co-crystal of puerarin and berberine hydrochloride involved in the present application comprises the following steps: (1) Puerarin and berberine hydrochloride raw materials are placed in a mortar in a molar ratio of 1:0.9-1:1.1, and are ground uniformly to obtain a mixture; (2) The mixture obtained in step (1) is ground in a clockwise direction, and 100 μL of a solvent is added dropwise thereto every 15 min, and is ground for 2 h; (3) The product obtained in step (2) is dried to obtain a solid product in which puerarin and berberine hydrochloride form a co-crystal.
[0048] 2.2 The organic solvent in the preparation method of the co-crystal of puerarin and berberine hydrochloride involved in the present application is preferably a mixed solvent prepared by different proportions of any one or more of ethanol, acetonitrile, isopropanol and methanol.
[0049] 2.3 The mixed solid substance containing the co-crystal of puerarin and berberine hydrochloride of the present application is prepared by mixing the co-crystal of puerarin and berberine hydrochloride obtained by the above method with other chemical substances in any non-zero ratio and by a conventional method.
[0050] 3. Pharmaceutical composition, dosage feature and pharmaceutical use of the co-crystal of puerarin and berberine hydrochloride: 3.1 The pharmaceutical composition of the present application contains the co-crystal of puerarin and berberine hydrochloride and a pharmaceutically acceptable carrier.
[0051] 3.2 The pharmaceutical composition of the present application contains the mixed solid substance of the co-crystal of puerarin and berberine hydrochloride and a pharmaceutically acceptable carrier.
[0052] 3.3 The pharmaceutical composition of the present application contains the co-crystal of puerarin and berberine hydrochloride in a daily dosage of 5-3000 mg.
[0053] 3.4 The pharmaceutical composition of the present application is characterized in that it is various tablets, capsules, pills, injection preparations, granules, powders, pellets, dripping pills, suppositories, film preparations, patches, aerosol preparations, spray preparations, sustained-release preparations or controlled-release preparations.
[0054] 3.5 The co-crystal of puerarin and berberine hydrochloride, the mixed solid substance containing the co-crystal of puerarin and berberine hydrochloride or the pharmaceutical composition of the present application are used for preparing a medicine for preventing and treating inflammation, cardiovascular system and diabetes.
[0055] The present application relates to a pharmaceutical composition containing the co-crystal of puerarin and berberine hydrochloride of the present application as an active ingredient. The pharmaceutical composition can be prepared according to the methods known in the art. The co-crystal of puerarin and berberine hydrochloride of the present application can be combined with one or more pharmaceutically acceptable solid or liquid excipients and / or adjuvants to form any dosage form suitable for human or animal use. The content of the co-crystal of puerarin and berberine hydrochloride of the present application in the pharmaceutical composition thereof is in the range of 10-90% by weight.
[0056] The co-crystal of puerarin and berberine hydrochloride of the present application can be administered in unit dosage form, and the administration route can be enteral or parenteral, mainly oral.
[0057] The administration dosage form of the present application is preferably a solid dosage form. The solid dosage form can be tablets (including ordinary tablets, enteric-coated tablets, buccal tablets, dispersible tablets, chewable tablets, effervescent tablets, oral disintegrating tablets), capsules (including hard capsules, soft capsules, enteric-coated capsules), granules, powders, pellets, dripping pills, suppositories, film preparations, patches, aerosol (powder) preparations, spray preparations, etc.
[0058] The co-crystal of puerarin and berberine hydrochloride of the present application can be prepared into common preparations, sustained-release preparations, controlled-release preparations, targeted preparations and various microparticle drug delivery systems.
[0059] In order to prepare the co-crystal of puerarin and berberine hydrochloride of the present application into tablets, various excipients known in the art can be widely used, including diluents, binders, wetting agents, disintegrants, lubricants, glidants. The diluents can be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc.; the wetting agents can be water, ethanol, isopropyl alcohol, etc.; the binders can be starch paste, dextrin, sugar 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.; the 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.; the lubricants and glidants can be talc, silicon dioxide, stearate, tartaric acid, liquid paraffin, polyethylene glycol, etc.
[0060] The tablets can also be further prepared into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double-layer tablets and multi-layer tablets.
[0061] In order to prepare the drug delivery unit into capsules, the effective ingredient, the co-crystal of puerarin and berberine hydrochloride of the present application, can be mixed with diluents and glidants, and the mixture can be directly placed into hard capsules or soft capsules. The effective ingredient, the co-crystal of puerarin and berberine hydrochloride of the present application, can also be first mixed with diluents, binders and disintegrants to prepare granules or pellets, and then placed into hard capsules or soft capsules. The various diluents, binders, wetting agents, disintegrants and glidants used for preparing the tablets of the co-crystal of puerarin and berberine hydrochloride of the present application can also be used for preparing the capsules of the co-crystal of puerarin and berberine hydrochloride of the present application.
[0062] In addition, if necessary, coloring agents, preservatives, flavors, taste correctors or other additives can also be added to the pharmaceutical preparations.
[0063] In order to achieve the purpose of medication and enhance the therapeutic effect, the pharmaceuticals of the present application can be administered by any known administration method.
[0064] The dosage of the pharmaceutical composition of the co-crystal of puerarin and berberine hydrochloride of the present 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, etc. The above-mentioned dosage can be administered as one dosage unit or divided into several dosage units, depending on the clinical experience of the doctor and the administration regimen including the use of other therapeutic means.
[0065] The co-crystal or composition of puerarin and berberine hydrochloride of the present application can be taken alone or in combination with other therapeutic drugs or symptomatic drugs. When the co-crystal of puerarin and berberine hydrochloride of the present application has a synergistic effect with other therapeutic drugs, its dosage should be adjusted according to the actual situation.
[0066] 4. The beneficial technical effects of the present application: the safety, solubility and bioavailability of the co-crystal of puerarin and berberine hydrochloride are superior, which is more beneficial to the in vivo absorption of drugs.
[0067] 4.1 The co-crystal of puerarin and berberine hydrochloride of the present application has no endothermic peak of losing crystallization solvent or crystallization water on its DSC spectrum ( Figure 6 ), and no weight loss peak before decomposition weight loss in its thermogravimetric analysis diagram ( Figure 7 ), indicating that the co-crystal of puerarin and berberine hydrochloride does not contain any crystallization solvent, and has the advantage of good safety of drug production.
[0068] 4.2 The co-crystal of puerarin and berberine hydrochloride of the present application shows better solubility than puerarin itself in water, especially the inherent dissolution rate has made significant progress ( Figure 9 ).
[0069] 4.3 The biological absorption effect of the drug and its pharmaceutical composition developed by the co-crystal of puerarin and berberine hydrochloride of the present application as an active ingredient after oral administration. The co-crystal of puerarin and berberine hydrochloride of the present application is used as an active ingredient. In the oral administration experiment using rats, the bioavailability of the co-crystal of puerarin and berberine hydrochloride is significantly better than that of puerarin or berberine hydrochloride ( Figure 10 ). The bioavailability of the co-crystal of puerarin and berberine hydrochloride of the present application is better than that of puerarin and berberine hydrochloride.
[0070] There are many factors affecting the dosage of the effective components of the co-crystal pharmaceutical composition of puerarin and berberine hydrochloride of the present application, such as: patient age, different body surface areas, different administration routes, different administration frequencies, different treatment purposes resulting in different dosages each time; different absorption and blood drug concentrations between samples, etc., also resulting in the suitable dosage range of the co-crystal composition of puerarin and berberine hydrochloride of the present application being 0.05-300 mg / kg body weight, preferably 0.1-50 mg / kg body weight. When used, different total dosage schemes of the effective components of the co-crystal of puerarin and berberine hydrochloride should be formulated according to the actual different treatment needs, and can be completed by multiple or one-time administration. BRIEF DESCRIPTION OF DRAWINGS
[0071] Figure 1 Molecular stereostructure ellipsograph of the co-crystal of puerarin and berberine hydrochloride.
[0072] Figure 2 Crystal cell packing diagram of the co-crystal of puerarin and berberine hydrochloride along the a axis.
[0073] Figure 3 Powder X-ray diffraction pattern of the co-crystal of puerarin and berberine hydrochloride.
[0074] Figure 4 Physical mixing powder X-ray diffraction pattern of the co-crystal of puerarin and berberine hydrochloride.
[0075] Figure 5 Infrared absorption spectrum of the co-crystal of puerarin and berberine hydrochloride.
[0076] Figure 6 Differential scanning calorimetry pattern of the co-crystal of puerarin and berberine hydrochloride.
[0077] Figure 7 Thermogravimetric pattern of the co-crystal of puerarin and berberine hydrochloride.
[0078] Figure 8 Differential scanning calorimetry and thermogravimetric superimposed pattern of puerarin, berberine hydrochloride and the co-crystal of puerarin and berberine hydrochloride.
[0079] Figure 9 Intrinsic dissolution rate pattern of the co-crystal of puerarin and berberine hydrochloride.
[0080] Figure 10 Pharmacokinetic pattern of the co-crystal of puerarin and berberine hydrochloride in rats.
[0081] Figure 11 Stability experiment pattern of the co-crystal of puerarin and berberine hydrochloride under high temperature, high humidity and light conditions. DETAILED DESCRIPTION
[0082] To better illustrate the technical solutions of the present application, the following examples are provided, but the present application is not limited to this.
[0083] Example 1 Preparation method 1 of the co-crystal of puerarin and berberine hydrochloride: Take puerarin and berberine hydrochloride in an appropriate amount according to a molar ratio of 1:1, dissolve in a single or mixed solvent such as ethanol / methanol / acetone / acetonitrile / isopropyl alcohol, completely dissolve the sample in a 50 °C water bath, and obtain single crystals after standing at room temperature for 7 days.
[0084] The obtained crystal sample is subjected to single crystal X-ray diffraction analysis, showing orthorhombic symmetry, and the space group is P 212121, the cell parameter value is a=7.65528 , b=15.07721, c=30.1929, α=90.00°, β=90.00°, γ=90.00°, V=3484.87, Z=4, indicating that the obtained sample is a co-crystal of puerarin and berberine hydrochloride.
[0085] Preparation method 2 of the co-crystal of puerarin and berberine hydrochloride: Take puerarin and berberine hydrochloride in an appropriate amount according to a molar ratio of 1:1, put them into a clean mortar, grind uniformly to obtain a physical mixture of puerarin and berberine hydrochloride, grind the obtained mixture in a clockwise direction, and add 100 μL of ethanol every 15 min, grind for 2 h, and filter, vacuum dry, filter natural dry or evaporative dry the obtained product. Subject it to powder X-ray diffraction analysis, and the diffraction spectrum is consistent with Figure 3 , indicating that the obtained sample is a co-crystal of puerarin and berberine hydrochloride.
[0086] Preparation method 3 of the co-crystal of puerarin and berberine hydrochloride: Take puerarin and berberine hydrochloride in an appropriate amount according to a molar ratio of 1:1, dissolve in a single or mixed solvent such as ethanol / methanol / acetone / acetonitrile / isopropyl alcohol, filter the clear solution while hot, rotary evaporate with a rotary evaporator, the pressure is 0.01 MPa, the water bath temperature is 45 °C, the rotation speed is 90 rpm, the time is 15 min, and the sample is dried under reduced pressure to obtain a co-crystal of puerarin and berberine hydrochloride. Subject it to powder X-ray diffraction analysis, and the diffraction spectrum is consistent with Figure 3 , indicating that the obtained sample is a co-crystal of puerarin and berberine hydrochloride.
[0087] Example 2 Stability test of the co-crystal of puerarin and berberine hydrochloride Light exposure test: 50 mg of the cocrystal of puerarin and berberine hydrochloride was placed under light conditions for 5 days and 10 days, and powder X-ray diffraction analysis was performed on it. The diffraction patterns were compared with... Figure 3 The consistency indicates that the sample is a eutectic of puerarin and berberine hydrochloride, which is stable under light conditions.
[0088] High-temperature test: 50 mg of the cocrystal of puerarin and berberine hydrochloride was placed under high-temperature conditions for 5 days and 10 days, and powder X-ray diffraction analysis was performed on it. The diffraction patterns were compared with... Figure 3 The consistency indicates that the sample is a eutectic of puerarin and berberine hydrochloride, which is stable under high temperature conditions.
[0089] High humidity test: 50 mg of the cocrystal of puerarin and berberine hydrochloride was placed under high humidity conditions for 5 days and 10 days, and powder X-ray diffraction analysis was performed on it. The diffraction patterns were compared with... Figure 3 The consistency indicates that the sample is a eutectic of puerarin and berberine hydrochloride, which is stable under high humidity conditions.
[0090] The above experimental results show that the cocrystal of puerarin and berberine hydrochloride has good stability under light, high temperature, and high humidity conditions, and is more stable than the anhydrous puerarin (crystalline) and hydrated berberine (crystalline) (see...). Figure 11 ).
[0091] Example 3 Solubility test Using puerarin (PUE) anhydrous (crystalline) and berberine hydrochloride (BER) hydrate (crystalline) as evaluation indicators, the content was determined by high-performance liquid chromatography (HPLC) at different wavelengths. The eutectic equilibrium solubility was calculated based on the puerarin content using the external standard method (see Table 3). The inherent dissolution rate curves are shown in Table 3. Figure 9 Based on the preliminary drug-drug synergistic design strategy, the cocrystal of puerarin and berberine hydrochloride significantly improved the solubility of puerarin while enhancing its own stability.
[0092] Table 3. Solubility curves of puerarin, berberine hydrochloride, and their eutectic in water.
[0093] Table 3 and Figure 9 Experimental data show that the cocrystal of puerarin and berberine hydrochloride has a faster dissolution rate and higher solubility. The dissolution rate of the cocrystal in water significantly improves the solubility of puerarin, allowing for rapid absorption and achieving effective blood drug concentrations. While improving the solubility of puerarin, it does not affect the in vivo absorption of berberine hydrochloride (see Example 4 and...). Figure 10 ).
[0094] Example 4 In vivo bioavailability test Standard solution: 1.00 mg of puerarin standard was precisely weighed, 1 mL of methanol was added respectively, and it was fully dissolved and mixed to prepare 1.00 mg / mL of puerarin stock solution, which was stored at -20°C. 1.00 mg of icariin was precisely weighed, 1 mL of methanol was added to fully dissolve and mix, and 1.00 mg / mL of icariin stock solution was prepared, which was stored at -20°C. 1.00 mg of berberine hydrochloride standard was precisely weighed, 1 mL of methanol was added respectively to fully dissolve and mix, and 1.00 mg / mL of berberine hydrochloride stock solution was prepared, which was stored at -20°C. The standard solution was diluted with methanol to different concentrations. Internal standard solution: 1.00 mg of propafenone was precisely weighed, 1 mL of methanol was added to fully dissolve and mix, and 1.00 mg / mL of propafenone stock solution was prepared, which was stored at -20°C. The internal standard working solution was diluted with methanol to 100 ng / mL.
[0095] SD rats were randomly divided into 4 groups, 8 rats in each group, and were raised under conventional feeding conditions. They were allowed to drink water freely, and were fasted for 12 h. Then, puerarin (PUE), berberine hydrochloride (BER), co-crystal of puerarin and berberine hydrochloride (PUE-BER), and simulated Gegen-Huanglian Decoction (s-GGHLD) were respectively administered to the rats at a dose of 20 mg / kg by gavage. At 0.05, 0.15, 0.3, 0.5, 1, 1.25, 1.5, 1.75, 2, 2.5, 3, 4, 5, 6, 8 and 24 h after administration, about 0.3 mL of blood was taken from the retro-orbital plexus, and was placed in a heparinized anticoagulant tube. The tube was centrifuged at 3000 rpm for 10 min at 4°C, and the supernatant was taken and stored in a -80°C freezer for testing.
[0096] For determination, 100 μL of plasma was precisely taken, 10 μL of internal standard propafenone working solution was added, and 890 μL of acetonitrile, a protein precipitant, was added. The mixture was fully shaken for 3 min, and was centrifuged (13000 rpm, 10 min). The upper solution was taken, and was dried by nitrogen blowing. Then, 100 μL of 50% methanol was added to re-dissolve the sample, which was fully shaken for 3 min, and was centrifuged (13000 rpm, 3 min). 90 μL of the upper solution was taken and was placed in an inner lining tube, and 10 μL of sample was injected. The standard curve was detected by the same method, and the peak area ratio of the drug to the internal standard was substituted into the standard curve to calculate the blood drug concentration of puerarin and berberine hydrochloride. The blood drug concentration-time curve was drawn.
[0097] Detection conditions: the chromatographic column was Acquity UPLC HSS T3 column (2.1 × 50 mm, 1.8 μm); the mobile phase A was 0.1% formic acid water, and the mobile phase B was acetonitrile. The flow rate was 0.25 mL / min. The column temperature was 30 °C. The injection volume was 10 μL. The gradient elution conditions were as follows: 15% B (0-0.5 min); 15%-40% B (0.5-2 min); 40%-80% B (2-4 min); 80%-90% B (4-5 min); 90%-10% B (5-7 min).
[0098] Mass spectrometry conditions: negative ion mode, puerarin mass-to-charge ratio was 417.1, and icariin mass-to-charge ratio was 675.2; positive ion mode, berberine hydrochloride mass-to-charge ratio was 336.0, and propafenone mass-to-charge ratio was 342.1.
[0099] The test results are shown in Table 4 and Figure 10 .
[0100] Table 4 Main pharmacokinetic parameters of puerarin, berberine hydrochloride, co-crystal and simulated puerarin and coptis soup (average ± standard error)
[0101] The above experimental research data show that, compared with the raw drug, the area under the drug-time curve of puerarin and berberine hydrochloride in the co-crystal AUC 0-24 increased by 3.0 and 1.4 times, respectively, and compared with the simulated traditional Chinese medicine compound, increased by 7.3 and 4.7 times, respectively; compared with the raw drug, the maximum blood drug concentration (Cmax) of puerarin and berberine hydrochloride in the co-crystal C max increased by 1.8 and 1.2 times, respectively, and compared with the simulated traditional Chinese medicine compound, increased by 10.2 and 3.6 times, respectively; the peak time (Tmax) of each group is not much different. T max
[0102] The in vivo bioavailability experimental results of Table 4 and Figure 10 show that the co-crystal of puerarin and berberine hydrochloride has a higher area under the drug-time curve, and an obvious double-peak absorption phenomenon, which is beneficial to the reabsorption of the drug, and the bioavailability is obviously better than that of the single drug and the simulated traditional Chinese medicine compound. This makes the co-crystal more easily reach the therapeutic concentration than any single raw drug used alone, and plays a synergistic role in disease treatment.
[0103] Example 5 Preparation method 1 (tablet) of combined drug preparation: Preparation method of the combined drug tablet: using the co-crystal of puerarin and berberine hydrochloride, using several excipients as the auxiliary ingredients for preparing the combined drug tablet, and preparing the tablet sample containing 5-500 mg of the co-crystal per tablet according to a specific ratio, the tablet formulation ratio is shown in Table 5: Table 5 Preparation formulation of the co-crystal of puerarin and berberine hydrochloride combined drug tablet
[0104] The method for preparing the tablet formulation using the co-crystal of puerarin and berberine hydrochloride as the raw material is as follows: mixing several excipients and the raw material uniformly, directly compressing the tablet, or mixing the auxiliary materials, granulating by dry method, mixing the raw material uniformly, and then compressing the tablet.
[0105] Preparation method 2 (tablet) of the combined drug formulation: Preparation method of the combined drug tablet: using the co-crystal of puerarin and berberine hydrochloride, using several excipients as the auxiliary ingredients for preparing the combined drug tablet, and preparing the tablet sample containing 5-500 mg of the co-crystal per tablet according to a specific ratio, the tablet formulation ratio is shown in Table 6: Table 6 Preparation formulation of the co-crystal of puerarin and berberine hydrochloride combined drug tablet
[0106] The method for preparing the tablet formulation using the co-crystal of puerarin and berberine hydrochloride as the raw material is as follows: mixing several excipients and the raw material uniformly, adding an appropriate amount of 1% sodium hydroxymethyl cellulose solution, preparing the soft material, sieving and granulating, drying the wet granules, sieving and granulating, mixing magnesium stearate and talc uniformly, and compressing the tablet.
[0107] Preparation method 3 (capsule) of the combined drug formulation: Preparation method of the combined drug capsule: using the co-crystal of puerarin and berberine hydrochloride as the raw material, using several excipients as the auxiliary ingredients for preparing the combined drug capsule, and preparing the capsule sample containing 5-500 mg of the drug per tablet according to a specific ratio, the capsule formulation ratio is shown in Table 7: Table 7 Raw material and auxiliary material formulation of the co-crystal of puerarin and berberine hydrochloride combined drug capsule formulation
[0108] The method for preparing the capsule using the co-crystal of puerarin and berberine hydrochloride as the raw material is as follows: mixing several excipients and the raw material uniformly, adding an appropriate amount of 1% sodium hydroxymethyl cellulose solution, preparing the wet granules, drying and sieving, granulating, mixing magnesium stearate uniformly, and inserting the capsule; or without the granulation step, directly mixing the co-crystal of puerarin and berberine hydrochloride raw material and several excipients auxiliary materials uniformly, sieving, and then directly loading into the capsule.
[0109] Example 6 Dosage 1 (tablet) of the co-crystal combination drug of puerarin and berberine hydrochloride: The pharmaceutical composition developed using the co-crystal of puerarin and berberine hydrochloride as the pharmaceutically active ingredient, using the co-crystal of puerarin and berberine hydrochloride as the active ingredient of the drug, the daily dosage of 200 mg can be prepared into 100 mg ordinary tablets, 1 tablet each time, 2 times a day, or 200 mg tablets, 1 tablet each time, once a day.
[0110] Dosage 2 (capsule) of the co-crystal combination drug of puerarin and berberine hydrochloride: The pharmaceutical composition developed using the co-crystal of puerarin and berberine hydrochloride as the pharmaceutically active ingredient, using the co-crystal of puerarin and berberine hydrochloride as the active ingredient of the drug, the daily dosage of 500 mg can be prepared into 250 mg capsules, 1 capsule each time, 2 times a day, or 500 mg capsules, 1 capsule each time, once a day.
Claims
1. A eutectic of puerarin and berberine hydrochloride, wherein the molar ratio of puerarin to berberine hydrochloride is 1:1, using Cu... K α The powder X-ray diffraction pattern of the eutectic, radiated and expressed in 2θ angles, has the following characteristic peaks: 10.2±0.2°, 11.3±0.2°, 12.4±0.2°, 12.8±0.2°, 13.6±0.2°, 14.0±0.2°, 15.1±0.2°, 15.5±0.2°, 15.9±0.2°, 16.2±0.2°, 17.0±0.2°, 17.3±0.2°, 18.2±0.2°, 19.12±0.2°, 19.8±0.2°; Preferably, Cu is used K α The powder X-ray diffraction pattern of the eutectic, radiated and expressed at a 2θ angle, has the following characteristic peaks and peak area percentages: ； Preferably, Cu is used K α Radiation, expressed in 2θ angles, and the powder X-ray diffraction pattern of the eutectic exhibits the following characteristic peaks: 10.2±0.2°, 11.3±0.2°, 12.4±0.2°, 12.8±0.2°, 13.6±0.2°, 14.0±0.2°, 15.1±0.2°, 15.5±0.2°, 15.9±0.2°, 16.2±0.2°, 17.0±0.2°, 17.3±0.2°, 18.2±0.2°, 19.12±0.2°, 19 0.8±0.2°, 21.1±0.2°, 22.5±0.2°, 23.1±0.2°, 23.3±0.2°, 23.8±0.2°, 25.4±0.2°, 25.9±0.2°, 26.7±0.2°, 28.6±0.2°, 29.3±0.2°, 31.1±0.2°, 31.5±0.2°, 33.7±0.2°, 35.5±0.2°, 36.1±0.2°, 36.5±0.2°, 37.2±0.2°; Preferably, Cu is used K α The powder X-ray diffraction pattern of the eutectic, radiated and expressed at a 2θ angle, has the following characteristic peaks and peak area percentages: ； Preferably, Cu is used K α The powder X-ray diffraction pattern of the eutectic is shown in Figure 3, with the radiation expressed as a 2θ angle.
2. The eutectic as claimed in claim 1, wherein when analyzed using Fourier transform infrared spectroscopy, the infrared spectrum of the eutectic is within 3558 ± 2 cm⁻¹. -1 3345±2 cm -1 3240±2 cm -1 3132±2 cm -1 2946±2 cm -1 2899±2 cm -1 2838±2 cm -1 1621±2 cm -1 1590±2 cm -1 1509±2 cm -1 1486±2 cm -1 1439±2cm -1 1389±2 cm -1 1362±2 cm -1 1339±2 cm -1 1300±2 cm -1 1273±2 cm -1 1254±2 cm -1 1234±2 cm -1 1215±2 cm -1 1172±2 cm -1 1141±2 cm -1 1098±2 cm -1 1060±2 cm -1 1037±2 cm -1 1002±2 cm -1 936±2 cm -1 913±2 cm -1 885±2 cm -1 831±2 cm -1 792±2cm -1 750±2 cm -1 707±2 cm -1 626±2 cm -1 537±2 cm -1 506±2 cm -1 416 cm -1 Preferably, the infrared spectrum of the eutectic is shown in Figure 5.
3. The eutectic according to claim 1, wherein when analyzed by single-crystal X-ray diffraction, it exhibits orthorhombic crystal symmetry and a space group of [missing information]. P 212121, the cell parameter value is a = 7.65528. , b=15.07721, c=30.1929, α=90.00°, β=90.00°, γ=90.00°, V=3484.87, Z=4.
4. The eutectic according to claim 1, wherein in the differential scanning calorimetry spectrum of the eutectic, when the heating rate is 10°C per minute, there is a main endothermic peak at 251 ± 5°C and a secondary endothermic peak at 258 ± 5°C; preferably, the differential scanning calorimetry spectrum of the eutectic is shown in Figure 6.
5. A method for preparing the eutectic according to any one of claims 1-4, comprising the following methods: (1) Dissolve puerarin and berberine hydrochloride in an organic solvent at a molar ratio of 1:0.9 to 1:1.1, and let stand at room temperature to obtain the eutectic; preferably, completely dissolve in a 50 °C water bath and let stand at room temperature for 7 days; or (2) Puerarin and berberine hydrochloride are ground in a mortar at a molar ratio of 1:0.9 to 1:1.1, then an organic solvent is added and the mixture is ground again. After drying, the eutectic is obtained; or (3) Dissolve puerarin and berberine hydrochloride in an organic solvent at a molar ratio of 1:0.9 to 1:1.1, filter, and evaporate to obtain the eutectic.
6. The preparation method according to claim 5, wherein method (2) comprises the following steps: 1) Place puerarin and berberine hydrochloride in a mortar at a molar ratio of 1:0.9 to 1:1.1 and grind them evenly to obtain a mixture; 2) Grind the mixture obtained in step 1), adding organic solvent dropwise every 15 minutes, and grind for 2 hours to obtain the product; preferably, grinding is performed in a clockwise direction; 3) Dry the product obtained in step 2) to obtain a solid product, the eutectic of puerarin and berberine hydrochloride; Preferably, the organic solvent is selected from one or more of ethanol, acetonitrile, acetone, isopropanol, and methanol; preferably, the volume ratio between any two of the organic solvents is 1:10 to 10:
1.
7. A solid mixture comprising the eutectic of any one of claims 1-4, wherein the amount of the eutectic is 1-99.9% by weight, preferably 10-99.9% by weight, more preferably 50-99.9% by weight, and most preferably 85-99.9% by weight, based on the weight of the solid mixture.
8. A pharmaceutical composition comprising a preventive and / or therapeutically effective amount of any one of claims 1-4, or a solid mixture of claim 7, and a pharmaceutically acceptable carrier; preferably, the daily dose of the pharmaceutical composition is 5 to 3000 mg.
9. The pharmaceutical composition of claim 8, wherein the pharmaceutical composition is prepared as a tablet, capsule, pill, injection, granule, powder, suppository, film, patch, aerosol or spray; preferably, the pill is a micro-pill or drop pill; preferably, the pharmaceutical composition is prepared as a sustained-release formulation or controlled-release formulation.
10. Use of the eutectic of any one of claims 1-4, the solid mixture of claim 7, or the pharmaceutical composition of claim 8 or 9 in the preparation of a medicament for the prevention and / or treatment of inflammatory diseases, cardiovascular diseases, diabetes, bacterial dysentery, tuberculosis, scarlet fever, acute tonsillitis, or respiratory infections; preferably, the inflammatory disease is gastroenteritis.