Preparation method of traditional Chinese medicine meridian prescription drug pair supramolecular system

Abstract

The invention provides a preparation method of a traditional Chinese medicine prescription medicine pair supramolecular system, and belongs to the field of traditional Chinese medicines. The preparation method comprises the following steps: (1) extracting a traditional Chinese medicine meridian prescription pair by using a solvent, and taking an extracting solution; and (2) recovering the solvent from the extracting solution, concentrating to obtain extract liquid, enabling the active components to be self-assembled to generate supramolecular system precipitate, standing, filtering, and taking the precipitate, thereby obtaining the product. The extraction rate of active ingredients in the supramolecular system prepared by adopting a specific method is high, and the extraction rate of berberine, glycyrrhizic acid and other active ingredients in the supramolecular system prepared by adopting the specific method can reach 80% or above by taking a coptis chinensis-licorice drug pair as an example. The method is simple in technological process, economical and rapid, can be popularized and applied to the traditional Chinese medicine industry, realizes industrial production of a supramolecular system, and is beneficial to reasonable research and development of traditional Chinese medicine prescriptions.

Description

Technical Field

[0001] The invention belongs to the field of traditional Chinese medicine, and in particular relates to a method for preparing a supramolecular system of a traditional Chinese medicine prescription. Background Art

[0002] Drug pairs are a common way of combining traditional Chinese medicines. During the decoction process, the active ingredients of many traditional Chinese medicine pairs will form supramolecular systems, which have an important impact on the properties, taste and efficacy of traditional Chinese medicines (Huang XM, Wang PL, Li T, et al. Self-assemblies based on traditional medicine berberine and cinnamic acid for adhesion-induced inhibition multidrug-resistant Staphylococcus aureus [J]. ACS Appl Mater Interfaces, 2020, 12: 227-237.). The coptis chinensis-licorice herb pair is an important component of classic prescriptions such as Huanglian Decoction and Licorice Decoction for Xiexin. Modern research has found that more than 30% of the supramolecular system exists in the decoction of the coptis chinensis-licorice herbal pair, and the supramolecular system in the decoction was extracted by dialysis and freeze-drying methods, and the supramolecular system was characterized by scanning electron microscopy and dynamic light scattering. Through mouse diarrhea experiments, sensory evaluation and antibacterial experiments, it was confirmed from the perspective of "nature, taste and effect" that the supramolecular system formed after the combination can reduce the bitter and cold nature of coptis chinensis while enhancing its antibacterial activity. The material basis for the toxicity reduction and efficacy enhancement of licorice and coptis chinensis comes from the supramolecular system formed by the combination of the drug pair rather than the simple addition of the drug taste (Li Wen, Wang Zhijia, Lin Xiaoyu, et al. Exploration of the material basis of the "nature-taste-effect" of licorice and coptis chinensis based on the supramolecular system induced by weak bonds [J]. Acta Pharmaceutica Sinica, 2022, 57(6): 1901-1908.). The self-precipitation of Huanglian Jiedu Decoction has a definite effect of resisting neuronal cell damage and inhibiting neuronal cell apoptosis, which is better than the supernatant (Zhang C, Zhao R, Yan W, et al. Compositions, formation mechanism, and neuroprotective effect of compound precipitation from the traditional Chinese prescription Huang-Lian-Jie-Du-Tang [J]. Molecules, 2016, 21 (8): 1094.). A large number of studies have confirmed that the supramolecular system formed by the combination of traditional Chinese medicines is the material basis of the efficacy of traditional Chinese medicine prescriptions, reflecting the scientific connotation of the theory of Chinese medicine compatibility.

[0003] However, the external appearance of supramolecular systems of Chinese medicines is mostly in the form of self-precipitation of compound decoctions, which are often filtered out in the modern production process of Chinese medicine, resulting in loss of active ingredients, or their formation is avoided, thus losing the essence of traditional compatibility. The existing technology uses dialysis and freeze-drying methods to extract supramolecular systems from decoctions, but the dialysis and freeze-drying methods are costly, the extraction rate of active molecules is not high, and they are not suitable for industrial production. In addition, there are no other reports on the extraction and utilization of supramolecular systems. Summary of the invention

[0004] The purpose of the present invention is to provide a method for preparing a supramolecular system of a traditional Chinese medicine prescription.

[0005] The present invention provides a method for preparing a supramolecular system of a traditional Chinese medicine prescription, which comprises the following steps:

[0006] (1) extracting the Chinese herbal medicine pair using a solvent to obtain an extract;

[0007] (2) recovering the solvent from the extract, concentrating it to an extract solution, allowing the active ingredients to self-assemble to generate a supramolecular system precipitate, allowing it to stand for a while, filtering it, and taking the precipitate to obtain the extract solution.

[0008] Furthermore, in step (1), the Chinese herbal medicine pair is coptis chinensis-licorice herbal pair, scutellaria baicalensis-coptis chinensis herbal pair, kudzu root-coptis chinensis herbal pair, coptis chinensis-evodia rutaecarpa herbal pair, aconite-licorice herbal pair, aconite-rhubarb herbal pair.

[0009] Furthermore, in step (1), the Chinese herbal medicine pair is a coptis chinensis-licorice herbal pair; the mass ratio of coptis chinensis to licorice in the coptis chinensis-licorice herbal pair is 1:0.25-3.

[0010] Furthermore, the mass ratio of coptis chinensis to licorice in the coptis chinensis-licorice herb pair is 1:1.2-2;

[0011] Preferably, the mass ratio of Coptis chinensis to Licorice in the Coptis chinensis-Licorice herb pair is 1:2.

[0012] Further,

[0013] In step (1), the extraction method is to use solvent reflux to extract the traditional Chinese medicine prescription drug pair;

[0014] And / or, in step (2), the extract liquid is concentrated to a crude drug concentration of 1 g / mL;

[0015] And / or, in step (2), the standing is 4 to 50° C. for 3 to 70 hours;

[0016] And / or, in step (2), the precipitate is taken and then washed and dried.

[0017] Further,

[0018] In step (1), the number of reflux extraction is 1 to 3 times;

[0019] And / or, in step (1), the Chinese medicinal formula drug pair is soaked in a solvent before the reflux extraction;

[0020] And / or, in step (1), the solvent used in the reflux extraction is 8 to 10 times the amount of the traditional Chinese medicine prescription drug pair;

[0021] Preferably,

[0022] In step (1), the solvent is 75% ethanol aqueous solution;

[0023] And / or, in step (1), the reflux extraction time is 1 to 3 hours each time;

[0024] And / or, in step (1), the soaking time before the reflux extraction is 1 to 3 hours.

[0025] Further,

[0026] In step (1), the method of using a solvent to extract a Chinese herbal medicine pair comprises the following steps:

[0027] 1) Take a Chinese medicine prescription pair, add 8 to 10 times the amount of 75% ethanol aqueous solution, soak for 1 to 3 hours, and then reflux extraction for 1 to 3 hours;

[0028] 2) After step 1), reflux extraction is performed and filtered, 8 to 10 times the amount of 75% ethanol aqueous solution is added to the residue and refluxed for 1 to 3 hours, filtered, and the filtrates from the two reflux extractions are combined;

[0029] Preferably, the method of extracting a Chinese herbal medicine pair using a solvent comprises the following steps:

[0030] 1) Take a Chinese medicine pair, add 10 times the amount of 75% ethanol aqueous solution, soak for 2 hours, and then reflux extraction for 1 hour;

[0031] 2) Step 1) After reflux extraction, filter, add 8 times the amount of 75% ethanol aqueous solution to the residue and reflux extract for 1 hour, filter, and combine the filtrates from the two reflux extractions.

[0032] Further,

[0033] In step (2), the concentration is concentration under reduced pressure;

[0034] And / or, in step (2), the standing is 25-50° C. for 3-16 hours;

[0035] And / or, in step (2), the washing is washing with water;

[0036] And / or, in step (2), the drying is vacuum drying at 70°C;

[0037] Preferably,

[0038] In step (2), the temperature of the reduced pressure concentration is 70° C.

[0039] And / or, in step (2), the standing is standing at 50° C. for 3 h or standing at 25° C. for 16 h.

[0040] The present invention also provides a supramolecular system of a Chinese medicine prescription, which is prepared by the above-mentioned preparation method;

[0041] Preferably, the supramolecular system is prepared from the Coptis chinensis-Glycyrrhiza uralensis herb pair according to the aforementioned preparation method.

[0042] The present invention also provides the use of the aforementioned supramolecular system of traditional Chinese medicine prescriptions in the preparation of medicines;

[0043] Preferably, the drug is an anti-pathogenic microorganism, anti-gastric ulcer, or blood sugar-lowering drug.

[0044] In the prior art, dialysis and freeze-drying methods are often used to extract the supramolecular system in the decoction when preparing supramolecular systems of Chinese medicine drug pairs. However, the dialysis and freeze-drying methods are costly, have a long production cycle, have a low extraction rate of active molecules, and are not suitable for industrial large-scale production. In order to solve the problems existing in the production of supramolecular systems of Chinese medicine drug pairs in the prior art, a preparation method in the present invention is provided. The inventors continuously screened the process parameters in the preparation method and finally obtained a preparation method for a supramolecular system of a Chinese medicine prescription drug pair. The supramolecular system in the Coptis chinensis-Glycyrrhizic acid herb pair was extracted using the specific preparation method of the present invention. The extraction rate of berberine and glycyrrhizic acid, the active ingredients in the obtained supramolecular system, was significantly better than that of other preparation methods, and the effect was very excellent.

[0045] Compared with the prior art, the present invention has the following beneficial effects:

[0046] The present invention provides a method for preparing a supramolecular system of a Chinese medicine prescription. The present invention adopts a specific method to prepare a supramolecular system with a high extraction rate of active ingredients. Taking the coptis root-licorice herb pair as an example, the extraction rate of active ingredients such as berberine and glycyrrhizic acid in the supramolecular system prepared by the specific method can reach more than 80%. The process of the present invention is simple, economical and fast, and can be promoted for use in the Chinese medicine industry to realize the industrial production of supramolecular systems, which is beneficial to the rational research and development of Chinese medicine prescriptions.

[0047] The present invention takes the herbal pair of Coptis chinensis and Licorice as an example, and adopts specific process parameters to prepare a supramolecular system with high yield of active ingredients berberine and glycyrrhizic acid, which not only ensures the efficacy, but also solves the problem that free molecules such as berberine taste bitter and have poor patient adaptability, and greatly improves the drugability. At the same time, the preparation method of the present invention is simple and convenient, suitable for industrial large-scale production, and can be widely used in the production of pharmaceutical preparations such as anti-pathogenic microorganisms, anti-gastric ulcers, and hypoglycemic agents.

[0048] Obviously, according to the above contents of the present invention, in accordance with common technical knowledge and customary means in the art, without departing from the above basic technical ideas of the present invention, other various forms of modification, replacement or change may be made.

[0049] The above contents of the present invention are further described in detail below through specific implementation methods in the form of embodiments. However, this should not be understood as the scope of the above subject matter of the present invention being limited to the following examples. All technologies realized based on the above contents of the present invention belong to the scope of the present invention. DETAILED DESCRIPTION

[0050] The raw materials and equipment used in the specific embodiments of the present invention are all known products and are obtained by purchasing commercially available products.

[0051] Instruments and reagents

[0052] Agilent 1200 high performance liquid chromatography system, consisting of G1311A quaternary gradient pump, G1315D diode array detector, G1329A automatic sample injector, ChemStation high performance liquid chromatography chemical workstation; XS205DU electronic balance (Mettler-Toledo, Switzerland), UV-2401PC ultraviolet spectrophotometer (Shimadzu, Japan), Milli-Q ultrapure water system (Millipore), acetonitrile (chromatographic grade, Fisher), water was ultrapure water, and the other reagents were of analytical grade.

[0053] Berberine hydrochloride, glycyrrhizic acid, and liquiritin were purchased from the China Food and Drug Administration. Coptis chinensis Franch. and Glycyrrhiza uralensis Fisch. were purchased from the Chengdu Lotus Pond Medicinal Materials Market.

[0054] Example 1: Preparation method of supramolecular system of traditional Chinese medicine prescription

[0055] Coptis chinensis and licorice were prepared in a mass ratio of 1:2, with a total of 100 g of material added. Before extraction, 10 times the amount (1000 mL) of 75% ethanol aqueous solution was added to the residues for soaking at room temperature for 2 h, then refluxed for extraction for 1 h, filtered, and 8 times the amount (800 mL) of 75% ethanol aqueous solution was added to the residues for reflux extraction for 1 h, filtered, and the filtrate was combined. The ethanol was recovered under reduced pressure at 70°C until the remaining extract liquid was about 100 mL. The extract was allowed to stand at 25°C for 16 h, and the precipitate was filtered out. The precipitate was washed with a small amount of water, dried in a vacuum at 70°C, and weighed to obtain a supramolecular system.

[0056] When coptis chinensis and licorice are extracted separately, berberine and glycyrrhizic acid are free small molecules, which are clear and transparent in the extract and will not precipitate. When coptis chinensis and licorice are extracted together, the active ingredients such as berberine and glycyrrhizic acid will precipitate through non-covalent bond interactions to form a supramolecular system.

[0057] The obtained supramolecular system was tested according to the content determination method under Coptis chinensis and Licorice in the "Pharmacopoeia of the People's Republic of China" (2020 edition), and the yields of the main components berberine and glycyrrhizic acid were measured to be 84.6% and 94.2%, respectively.

[0058] Example 2: Preparation method of supramolecular system of traditional Chinese medicine prescription

[0059] Coptis chinensis and licorice were prepared in a mass ratio of 1:1.2, with a total of 100 g of material added. Before extraction, 10 times the amount (1000 mL) of 75% ethanol aqueous solution was added to the residues for soaking at room temperature for 2 h, then refluxed for extraction for 1 h, filtered, and 8 times the amount (800 mL) of 75% ethanol aqueous solution was added to the residues for reflux extraction for 1 h, filtered, and the filtrate was combined. The ethanol was recovered under reduced pressure at 70°C until the remaining extract liquid was about 100 mL. The extract was allowed to stand at 25°C for 16 h, and the precipitate was filtered out. The precipitate was washed with a small amount of water, dried in a vacuum at 70°C, and weighed to obtain a supramolecular system.

[0060] The obtained supramolecular system was tested according to the content determination method under Coptis chinensis and Licorice in the "Pharmacopoeia of the People's Republic of China" (2020 edition), and the yields of the main components berberine and glycyrrhizic acid were measured to be 78.4% and 91.4%, respectively.

[0061] Example 3: Preparation method of supramolecular system of traditional Chinese medicine prescription

[0062] Coptis chinensis and liquorice were prepared in a mass ratio of 1:2, with a total of 100 g of material added. Before extraction, 10 times the amount (1000 mL) of 75% ethanol aqueous solution was added to the residues for soaking at room temperature for 2 h, then refluxed for extraction for 1 h, filtered, and 8 times the amount (800 mL) of 75% ethanol aqueous solution was added to the residues for reflux extraction for 1 h, filtered, and the filtrate was combined. The ethanol was recovered under reduced pressure at 70°C until the remaining extract liquid was about 100 mL, and allowed to stand at 50°C for 3 h. The precipitate was filtered out, washed with a small amount of water, dried in a vacuum at 70°C, and weighed to obtain a supramolecular system.

[0063] The obtained supramolecular system was tested according to the content determination method under Coptis chinensis and Licorice in the "Pharmacopoeia of the People's Republic of China" (2020 edition), and the yields of the main components berberine and glycyrrhizic acid were measured to be 80.8% and 94.1%, respectively.

[0064] Example 4: Preparation method of supramolecular system of traditional Chinese medicine prescription

[0065] Coptis chinensis and licorice were prepared in a mass ratio of 1:1.2, with a total of 100 g of material added. Before extraction, 10 times the amount (1000 mL) of 75% ethanol aqueous solution was added to the residues for soaking at room temperature for 2 h, then refluxed for extraction for 1 h, filtered, and 8 times the amount (800 mL) of 75% ethanol aqueous solution was added to the residues for reflux extraction for 1 h, filtered, and the filtrate was combined. Ethanol was recovered under reduced pressure at 70°C until the remaining extract liquid was about 100 mL, and allowed to stand at 50°C for 3 h. The precipitate was filtered out, washed with a small amount of water, dried in a vacuum at 70°C, and weighed to obtain a supramolecular system.

[0066] The obtained supramolecular system was tested according to the content determination method under Coptis chinensis and Licorice in the "Pharmacopoeia of the People's Republic of China" (2020 edition), and the yields of the main components berberine and glycyrrhizic acid were measured to be 73.0% and 91.9%, respectively.

[0067] The beneficial effects of the present invention are demonstrated below through specific test examples.

[0068] Test Example 1: Screening of the Preparation Method of the Supramolecular System of the Traditional Chinese Medicine Prescription of the Present Invention

[0069] Methods and Results

[0070] 1.1 Content test method: Same as the content determination method under Coptis chinensis and Licorice in the Pharmacopoeia of the People's Republic of China (2020 edition).

[0071] 1.2 Calculation of extraction rate: Determine the content of glycyrrhizic acid and berberine in each extract and medicinal material according to the content determination method, and calculate the extraction rate according to the formula:

[0072] Extraction rate (%) = W t A t / WA×100%

[0073] W t is the weight of each extract, A t is the content of glycyrrhizic acid or berberine in each extract, W is the weight of the medicinal material, and A is the content of glycyrrhizic acid or berberine in the medicinal material.

[0074] The extraction rate (i.e., yield) of glycyrrhizic acid and berberine from the medicinal materials was calculated in each experiment.

[0075] 1.3 Preparation method screening

[0076] In the method for preparing the supramolecular system of the present invention, the four parameters of the ratio of medicinal materials (mass ratio of coptis root and liquorice), pH value, standing time and standing temperature are changed to screen the preparation method. Different selections of parameters are shown in Table 1. Each time, 100g of material is added according to the different ratios listed in Table 1, and 10 times (1000mL) of 75% ethanol aqueous solution is added before extraction to soak at room temperature for 2h, and then reflux extraction is performed for 1h, filtered, and the residue is then added with 8 times (800mL) of 75% ethanol aqueous solution for reflux extraction for 1h, filtered, and the filtrate is combined, and ethanol is recovered under reduced pressure at 70°C until the remaining extract liquid is about 100mL. The extract liquid and the precipitate therein are all transferred to a beaker, and the pH value is adjusted according to the conditions in Table 1 (the pH value of the extract liquid is about 4.7, and the pH value is adjusted by using NaOH aqueous solution or HCl aqueous solution), standing, filtering the precipitate, washing the precipitate with a small amount of water, vacuum drying at 70°C, weighing the weight, and obtaining the supramolecular system. The supramolecular system is ground for use and the content is determined. The experimental results are shown in Table 1.

[0077] Table 1. Process parameter selection and results

[0078]

[0079] The extraction rates of berberine and glycyrrhizic acid in the supramolecular system prepared by various process parameters are shown in Table 1. It can be seen from Table 1 that when the mass ratio of coptis chinensis: licorice is 1:2 and 1:1.2, the pH value is not adjusted (the pH of the extract is about 4.7), and the berberine and glycyrrhizic acid extraction rates in the supramolecular system are higher than those of other process parameters. Among them, the extraction rate of active ingredients obtained by the mass ratio of coptis chinensis: licorice is 1:2, the pH value is not adjusted (about pH 4.7), and the extraction rate is about 16 hours at room temperature (about 25°C).

[0080] In summary, the present invention provides a method for preparing a supramolecular system of a Chinese medicine prescription. The present invention adopts a specific method to prepare a supramolecular system with a high extraction rate of active ingredients. Taking the coptis chinensis-licorice herb pair as an example, the extraction rate of active ingredients such as berberine and glycyrrhizic acid in the supramolecular system prepared by the specific method can reach more than 80%. In addition, the process of the present invention is simple, economical and fast, and can be promoted for use in the Chinese medicine industry to realize the industrial production of supramolecular systems, which is conducive to the rational research and development of Chinese medicine prescriptions.

Claims

1. A method for preparing a supramolecular system of a Chinese medicine prescription, characterized in that: It includes the following steps: (1) extracting the Chinese herbal medicine pair using a solvent to obtain an extract; (2) recovering the solvent from the extract, concentrating it to an extract solution, allowing the active ingredients to self-assemble to generate a supramolecular system precipitate, allowing it to stand for a while, filtering it, and taking the precipitate to obtain the extract solution.

2. The preparation method according to claim 1, characterized in that: In step (1), the Chinese herbal medicine pairs are coptis chinensis-licorice herbal pair, scutellaria baicalensis-coptis chinensis herbal pair, kudzu root-coptis chinensis herbal pair, coptis chinensis-evodia rutaecarpa herbal pair, aconite-licorice herbal pair, aconite-rhubarb herbal pair.

3. The preparation method according to claim 2, characterized in that: In step (1), the Chinese herbal medicine pair is a coptis chinensis-licorice herbal medicine pair; the mass ratio of coptis chinensis to licorice in the coptis chinensis-licorice herbal medicine pair is 1:0.25-3.

4. The preparation method according to claim 3, characterized in that: The mass ratio of coptis chinensis to licorice in the coptis chinensis-licorice herb pair is 1:1.2-2; Preferably, the mass ratio of Coptis chinensis to Licorice in the Coptis chinensis-Licorice herb pair is 1:

2.

5. The preparation method according to claim 1, characterized in that: In step (1), the extraction method is to use solvent reflux to extract the traditional Chinese medicine prescription drug pair; And / or, in step (2), the extract liquid is concentrated to a crude drug concentration of 1 g / mL; And / or, in step (2), the standing is 4 to 50° C. for 3 to 70 hours; And / or, in step (2), the precipitate is taken and then washed and dried.

6. The preparation method according to claim 5, characterized in that: In step (1), the number of reflux extraction is 1 to 3 times; And / or, in step (1), the Chinese medicinal formula drug pair is soaked in a solvent before the reflux extraction; And / or, in step (1), the solvent used in the reflux extraction is 8 to 10 times the amount of the traditional Chinese medicine prescription drug pair; Preferably, In step (1), the solvent is 75% ethanol aqueous solution; And / or, in step (1), the reflux extraction time is 1 to 3 hours each time; And / or, in step (1), the soaking time before the reflux extraction is 1 to 3 hours.

7. The preparation method according to claim 6, characterized in that: In step (1), the method of using a solvent to extract a Chinese herbal medicine pair comprises the following steps: 1) Take a Chinese medicine prescription pair, add 8 to 10 times the amount of 75% ethanol aqueous solution, soak for 1 to 3 hours, and then reflux extraction for 1 to 3 hours; 2) After step 1), reflux extraction and filtration, add 8 to 10 times the amount of 75% ethanol aqueous solution to the residue and reflux extract for 1 to 3 hours, filter, and combine the filtrates from the two reflux extractions; Preferably, the method of extracting a Chinese herbal medicine pair using a solvent comprises the following steps: 1) Take a Chinese medicine pair, add 10 times the amount of 75% ethanol aqueous solution, soak for 2 hours, and then reflux extraction for 1 hour; 2) Step 1) After reflux extraction, filter, add 8 times the amount of 75% ethanol aqueous solution to the residue and reflux extract for 1 hour, filter, and combine the filtrates from the two reflux extractions.

8. The preparation method according to claim 5, characterized in that: In step (2), the concentration is concentration under reduced pressure; And / or, in step (2), the standing is 25-50° C. for 3-16 hours; And / or, in step (2), the washing is washing with water; And / or, in step (2), the drying is vacuum drying at 70°C; Preferably, In step (2), the temperature of the reduced pressure concentration is 70° C. And / or, in step (2), the standing is standing at 50° C. for 3 h or standing at 25° C. for 16 h.

9. A supramolecular system of a Chinese medicine prescription, characterized in that: It is prepared by the preparation method described in any one of claims 1 to 8; Preferably, the supramolecular system is prepared from the Coptis chinensis-Glycyrrhiza uralensis herb pair according to the preparation method according to any one of claims 1 to 8.

10. Use of the supramolecular system of the traditional Chinese medicine prescription drug pair according to claim 9 in the preparation of medicines; Preferably, the drug is an anti-pathogenic microorganism, anti-gastric ulcer, or blood sugar-lowering drug.

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