Traditional Chinese medicine extract with cold dispelling effect, preparation method of traditional Chinese medicine extract and application of traditional Chinese medicine extract in preparation of rheumatoid arthritis medicine

Abstract

The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a traditional Chinese medicine extract with a cold dispelling effect, a preparation method thereof and application of the traditional Chinese medicine extract in preparation of rheumatoid arthritis medicines. The preparation method comprises the following steps: adding water into cassia twig and ginger, heating, refluxing, and filtering to obtain volatile oil, filtrate and decoction dregs 1; heating and refluxing the filtrate, Chinese herbaceous peony, liquorice, ephedra and an ethanol water solution to obtain an extracting solution 2 and a medicine residue 2, and concentrating the extracting solution 2 to obtain an extract 1; rhizoma atractylodis macrocephalae, rhizoma anemarrhenae, radix saposhnikoviae, radix aconiti carmichaeli praeparata, the medicine residue 1, the medicine residue 2 and the solvent 1 are subjected to heating reflux, an extracting solution 2 and a medicine residue 3 are obtained, and the extracting solution 2 is concentrated to obtain an extract 2; carrying out ultrasonic extraction on the medicine residue 3, mixed enzyme and a solvent 2 to obtain an extracting solution 3, and concentrating the extracting solution 3 to obtain an extract 3; and uniformly mixing the volatile oil, the extract 1, the extract 2 and the extract 3 to obtain the traditional Chinese medicine extract. Compared with other extraction methods, the prepared traditional Chinese medicine extract has remarkable effects of dispelling cold and treating rheumatoid arthritis.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a traditional Chinese medicine extract with cold-dispelling effect, a preparation method thereof and an application of the extract in preparing a medicine for rheumatoid arthritis. Background Art

[0002] Guizhi Shaoyao Zhimu Decoction is a prescription from the Synopsis of the Golden Chamber written by Zhang Zhongjing, the medical saint of the Han Dynasty. It is composed of cinnamon twig, peony root, licorice, ephedra, ginger, white atractylodes, anemarrhena, saposhnikovia, and aconite. It is mainly used to treat pain in limbs, weakness, swollen feet, dizziness, shortness of breath, nausea, red tongue with white fur, and weak pulse. It is mainly used to treat rheumatoid arthritis, rheumatic arthritis, polymyalgia rheumatica, fibromyalgia, sciatica, femoral head necrosis, tenosynovitis and other diseases. It can also be used for measles complicated with pneumonia, tracheitis, cor pulmonale with heart failure, deep tissue inflammation and other diseases.

[0003] However, Guizhi Shaoyao Zhimu Decoction is a decoction, which requires a large dosage, is inconvenient to take, and has unpleasant tastes such as bitter, sour, astringent and spicy, resulting in poor patient compliance. In addition, it is difficult to store and is prone to mold and deterioration.

[0004] Improve the Guizhi Shaoyao Zhimu Decoction, and prepare it in the form of an extract, which is convenient for storage. For example, the Chinese patent with the authorization publication number CN110638990B discloses a method for extracting the active ingredients of the Guizhi Shaoyao Zhimu prescription preparation, which includes the following steps: 1) taking the prescription pieces of the Guizhi Shaoyao Zhimu prescription preparation, crushing, and mixing; 2) adding 12-16 times the amount of water for extraction, the extraction time is 1-2h, and the number of extractions is 3-4 times. The technical scheme uses the paste rate, cinnamaldehyde in cinnamon twigs, paeoniflorin in white peony, mangoside in Zhimu, glycyrrhizic acid in licorice, and inflammatory factor IL-6 as the evaluation indicators of the extraction process, and uses the information entropy weighting method to determine the objective weight of each indicator, and conducts a comprehensive evaluation to obtain the optimal extraction process of the Guizhi Shaoyao Zhimu prescription preparation. The process is stable and feasible, with good repeatability, and the prepared extract has a high content of active ingredients, a significant anti-inflammatory effect, and can be used for mass production.

[0005] Zeng Hairong et al. also published a paper titled "Optimization of the Extraction Process of Guizhi Shaoyao Zhimu Granules Based on Entropy Weight Method Combined with Box-Behnken Response Surface Method" in the Journal of Chinese Herbal Medicine. The study showed that the optimal extraction process conditions for Guizhi Shaoyao Zhimu Granules were to add 16 times the amount of water and extract three times, each time for 1 hour.

[0006] However, using only water extraction may lead to the waste of some active ingredients in the raw materials of Guizhi Shaoyao Zhimu Decoction. Summary of the invention

[0007] In view of the above problems, the present invention provides a Chinese medicinal extract with the effect of expelling cold, a preparation method thereof and an application of the extract in preparing a drug for rheumatoid arthritis.

[0008] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0009] The first aspect of the present invention provides a method for preparing a Chinese herbal extract having a cold-dispelling effect, comprising the following steps:

[0010] Step S1, crushing cinnamon twig and ginger, soaking them in water, heating them under reflux, filtering them, and obtaining volatile oil, filtrate and medicinal residue 1;

[0011] Step S2, mixing the filtrate, peony root, liquorice, ephedra and ethanol aqueous solution, heating to reflux, filtering to obtain extract 2 and residue 2, and concentrating the extract 2 to obtain extract 1;

[0012] Step S3, mixing Atractylodes macrocephala, Rhizoma Anemarrhenae, Radix Saposhnikoviae, Radix Aconiti Lateralis Preparata, dregs 1, dregs 2 and solvent 1, heating and refluxing, filtering to obtain extract 2 and dregs 3, and concentrating extract 2 to obtain extract 2;

[0013] Step S4, mixing the drug residue 3, the mixed enzyme and the solvent 2, performing ultrasonic extraction, filtering and obtaining an extract 3, and concentrating the extract 3 to obtain an extract 3;

[0014] Step S5, mixing the volatile oil, extract 1, extract 2 and extract 3 evenly to obtain a Chinese medicine extract;

[0015] The solvent 1 in step S3 includes water, caprylic acid capric acid macrogol glyceride and castor oil polyoxyethylene ether;

[0016] The solvent 2 in step S4 includes water, betaine and 1,3-propylene glycol.

[0017] Preferably, the raw materials of the traditional Chinese medicine extract, by mass, include: 12 parts of cinnamon twig, 9 parts of peony root, 6 parts of licorice, 12 parts of ephedra, 15 parts of ginger, 15 parts of atractylodes macrocephala, 12 parts of anemarrhena, 12 parts of siler, and 10 parts of processed aconite.

[0018] In some preferred embodiments, the amount of water added in step S1 is 5-10 times the total mass of cassia twig and ginger, preferably 8 times.

[0019] In some preferred embodiments, the soaking temperature in step S1 is 25-30° C., and the soaking time is 1-2 h.

[0020] Preferably, the soaking temperature in step S1 is 25° C. and the soaking time is 2 hours.

[0021] In some preferred embodiments, the heating reflux temperature in step S1 is 85-95° C., and the heating reflux time is 3-7 h.

[0022] Preferably, the heating reflux temperature in step S1 is 90° C., and the heating reflux time is 4 hours.

[0023] In some preferred embodiments, the mass concentration of the ethanol aqueous solution in step S2 is 60-80%, preferably 75%.

[0024] In some preferred embodiments, the amount of ethanol aqueous solution added in step S2 is 4-8 times, preferably 6 times, the total mass of peony, licorice and ephedra.

[0025] In some preferred embodiments, the heating reflux temperature in step S2 is 75-85° C. and the time is 3-5 h.

[0026] Preferably, the heating reflux temperature in step S2 is 80° C. and the time is 4 hours.

[0027] Preferably, the concentration in step S2 is specifically: concentrating to 2-3 times of the extract 2, preferably 2 times.

[0028] In some preferred embodiments, the solvent 1 in step S3 comprises water, caprylic acid capric acid macrogol glyceride and castor oil polyoxyethylene ether in a mass ratio of 25-35:2-5:1.

[0029] Preferably, the solvent 1 in step S3 comprises water, caprylic acid capric acid macrogol glyceride and castor oil polyoxyethylene ether in a mass ratio of 30:3:1.

[0030] In some preferred embodiments, the heating reflux temperature in step S3 is 80-90° C. and the time is 2-5 h.

[0031] Preferably, the heating reflux temperature in step S3 is 85° C. and the time is 3 h.

[0032] In some preferred embodiments, the amount of solvent 1 added in step S3 is 5-10 times, preferably 8 times, the total mass of Atractylodes macrocephala, Rhizoma Anemarrhenae, Radix Saposhnikoviae, Radix Aconitum, dregs 1 and dregs 2.

[0033] Preferably, the concentration in step S3 is specifically: concentrating to 2-3 times of the extract 3, preferably 2 times.

[0034] In some preferred embodiments, the mixed enzyme in step S4 comprises β-glucosidase, cellulase and pectinase in a mass ratio of 1-2:3-7:1-2.

[0035] Preferably, the enzyme mixture in step S4 comprises β-glucosidase, cellulase and pectinase in a mass ratio of 1.5:5:1.

[0036] In some preferred embodiments, the solvent 2 in step S4 comprises water, betaine and 1,3-propylene glycol in a mass ratio of 4-8:1:1.5-2.5.

[0037] Preferably, the solvent 2 in step S4 comprises water, betaine and 1,3-propylene glycol in a mass ratio of 6:1:2.

[0038] In some preferred embodiments, the preparation method of the solvent 2 in step S4 is: betaine and 1,3-propylene glycol are stirred at 70-90° C. and 300-500 rpm for 10-15 min, and water is added to obtain the solvent 2.

[0039] Preferably, the preparation method of the solvent 2 in step S4 is: betaine and 1,3-propylene glycol are stirred at 80° C. and 400 rpm for 12 min, and water is added to obtain the solvent 2.

[0040] In some preferred embodiments, the mass ratio of the medicinal residue 3, the mixed enzyme and the solvent 2 in step S4 is 1:0.04-0.08:4-8.

[0041] Preferably, in step S4, the mass ratio of the medicinal residue 3, the mixed enzyme and the solvent 2 is 1:0.06:6.

[0042] In some preferred embodiments, the temperature of the ultrasonic extraction in step S4 is 25-30° C., and the time of the ultrasonic extraction is 1-3 h.

[0043] Preferably, the temperature of the ultrasonic extraction in step S4 is 25° C., and the time of the ultrasonic extraction is 2 h.

[0044] The second aspect of the present invention provides a Chinese herbal medicine extract with the effect of dispelling cold obtained by the above preparation method.

[0045] The third aspect of the present invention provides the use of the Chinese medicinal extract with the cold-dispelling effect obtained by the above preparation method in the preparation of rheumatoid arthritis drugs.

[0046] Compared with the prior art, the present invention has the following beneficial effects:

[0047] The present invention extracts medicinal materials in batches based on the components of Guizhi Shaoyao Zhimu Decoction, and adopts specific solvent 1, solvent 2 and mixed enzyme. Compared with other extraction methods, the prepared traditional Chinese medicine extract has significant effects of dispelling cold and treating rheumatoid arthritis. DETAILED DESCRIPTION

[0048] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0049] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0050] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0051] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0052] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0053] Exemplary manufacturers of the raw materials used in the following examples and comparative examples are as follows:

[0054] Caprylic / capric acid macrogol glycerides were purchased from Gattefossé (Shanghai) Trading Co., Ltd. under the trade name: Caprylic / capric macrogol glycerides;

[0055] Castor oil polyoxyethylene ether was purchased from Xingtai Lantian Fine Chemical Co., Ltd., model: EL-30;

[0056] β-Glucosidase was purchased from Beijing Biolab Technology Co., Ltd., catalog number: QN0491;

[0057] Cellulase was purchased from Beijing Biolab Technology Co., Ltd., catalog number: QN0504;

[0058] Pectinase was purchased from Beijing Biolab Technology Co., Ltd., catalog number: LA14000;

[0059] Neutral protease was purchased from Shanghai Maokang Biotechnology Co., Ltd., product number: MX1006-1000MG;

[0060] Betaine was purchased from Yisheng Biotechnology (Shanghai) Co., Ltd., product number: 60391ES60;

[0061] Choline chloride was purchased from Tieling Bright Biotechnology Co., Ltd.;

[0062] Stearamidopropyl hydroxypropyl sulfobetaine was purchased from Shandong Pioneer New Materials Co., Ltd.;

[0063] Palm kernel oleamidopropyl betaine was purchased from Tianjin Alta Technology Co., Ltd., product number: 1ST175419;

[0064] Neutral protease was purchased from Beijing Biolab Technology Co., Ltd., catalog number: KD3672;

[0065] Xylanase was purchased from Beijing Biolab Technology Co., Ltd., catalog number: TD0719;

[0066] Example 1

[0067] The Chinese herbal medicine extract with the effect of dispelling cold is composed of 12 parts of cinnamon twig, 9 parts of peony root, 6 parts of liquorice, 12 parts of ephedra, 15 parts of ginger, 15 parts of atractylodes macrocephala, 12 parts of anemarrhena, 12 parts of siler, and 10 parts of processed aconite.

[0068] The preparation method of the Chinese herbal extract with the effect of dispelling cold is as follows:

[0069] Step S1, crushing cinnamon twig and ginger, soaking them in water, heating them under reflux, filtering them, and obtaining volatile oil, filtrate and medicinal residue 1;

[0070] Step S2, mixing the filtrate, peony root, liquorice, ephedra and ethanol aqueous solution, heating to reflux, filtering to obtain extract 2 and residue 2, and concentrating the extract 2 to obtain extract 1;

[0071] Step S3, mixing Atractylodes macrocephala, Rhizoma Anemarrhenae, Radix Saposhnikoviae, Radix Aconiti Lateralis Preparata, dregs 1, dregs 2 and solvent 1, heating and refluxing, filtering to obtain extract 2 and dregs 3, and concentrating extract 2 to obtain extract 2;

[0072] Step S4, mixing the drug residue 3, the mixed enzyme and the solvent 2, performing ultrasonic extraction, filtering and obtaining an extract 3, and concentrating the extract 3 to obtain an extract 3;

[0073] Step S5, the volatile oil, extract 1, extract 2 and extract 3 are mixed evenly to obtain the Chinese medicine extract.

[0074] The amount of water added in step S1 is 5 times the total mass of cassia twig and ginger.

[0075] The soaking temperature in step S1 is 25° C. and the soaking time is 1 hour.

[0076] The heating reflux temperature in step S1 is 95° C., and the heating reflux time is 3 h.

[0077] The mass concentration of the ethanol aqueous solution in step S2 is 60%.

[0078] The amount of ethanol aqueous solution added in step S2 is 4 times the total mass of peony, liquorice and ephedra.

[0079] The heating reflux temperature in step S2 is 75° C. and the time is 5 h.

[0080] The concentration in step S2 specifically includes: concentrating to twice the amount of extract 2.

[0081] The solvent 1 in step S3 is composed of water, caprylic acid capric acid macrogol glyceride and castor oil polyoxyethylene ether in a mass ratio of 25:2:1.

[0082] The heating reflux temperature in step S3 is 80° C. and the time is 5 h.

[0083] The amount of solvent 1 added in step S3 is 5 times the total mass of Atractylodes macrocephala, Rhizoma Anemarrhenae, Radix Saposhnikoviae, Radix Aconitum, dregs 1 and dregs 2.

[0084] The concentration in step S3 specifically includes: concentrating to twice the volume of extract 3.

[0085] The mixed enzyme in step S4 consists of β-glucosidase, cellulase and pectinase in a mass ratio of 1:3:1.

[0086] The solvent 2 in step S4 is composed of water, betaine and 1,3-propylene glycol in a mass ratio of 4:1:1.5.

[0087] The preparation method of the solvent 2 in step S4 is as follows: betaine and 1,3-propylene glycol are stirred at 70° C. and 300 rpm for 15 min, and water is added to obtain the solvent 2.

[0088] The mass ratio of the medicinal residue 3, the mixed enzyme and the solvent 2 in step S4 is 1:0.04:4.

[0089] The temperature of the ultrasonic extraction in step S4 is 25° C., and the time of the ultrasonic extraction is 3 h.

[0090] Example 2

[0091] The Chinese herbal medicine extract with the effect of dispelling cold has the same raw material composition as that in Example 1.

[0092] The preparation method of the Chinese herbal extract with the effect of dispelling cold is as follows:

[0093] Step S1, crushing cinnamon twig and ginger, soaking them in water, heating them under reflux, filtering them, and obtaining volatile oil, filtrate and medicinal residue 1;

[0094] Step S2, mixing the filtrate, peony root, liquorice, ephedra and ethanol aqueous solution, heating to reflux, filtering to obtain extract 2 and residue 2, and concentrating the extract 2 to obtain extract 1;

[0095] Step S3, mixing Atractylodes macrocephala, Rhizoma Anemarrhenae, Radix Saposhnikoviae, Radix Aconiti Lateralis Preparata, dregs 1, dregs 2 and solvent 1, heating and refluxing, filtering to obtain extract 2 and dregs 3, and concentrating extract 2 to obtain extract 2;

[0096] Step S4, mixing the drug residue 3, the mixed enzyme and the solvent 2, performing ultrasonic extraction, filtering and obtaining an extract 3, and concentrating the extract 3 to obtain an extract 3;

[0097] Step S5, the volatile oil, extract 1, extract 2 and extract 3 are mixed evenly to obtain the Chinese medicine extract.

[0098] The amount of water added in step S1 is 10 times the total mass of cassia twig and ginger.

[0099] The soaking temperature in step S1 is 30° C. and the soaking time is 1 hour.

[0100] The heating reflux temperature in step S1 is 95° C., and the heating reflux time is 3 h.

[0101] The mass concentration of the ethanol aqueous solution in step S2 is 80%.

[0102] The amount of ethanol aqueous solution added in step S2 is 8 times the total mass of peony, liquorice and ephedra.

[0103] The heating reflux temperature in step S2 is 85° C. and the time is 3 h.

[0104] The concentration in step S2 specifically includes: concentrating to 3 times of the extract 2.

[0105] The solvent 1 in step S3 is composed of water, caprylic acid capric acid macrogol glyceride and castor oil polyoxyethylene ether in a mass ratio of 35:5:1.

[0106] The heating reflux temperature in step S3 is 90° C. and the time is 2 h.

[0107] The amount of solvent 1 added in step S3 is 10 times the total mass of Atractylodes macrocephala, Rhizoma Anemarrhenae, Radix Saposhnikoviae, Radix Aconitum, dregs 1 and dregs 2.

[0108] The concentration in step S3 specifically includes: concentrating to 3 times of the extract 3.

[0109] The mixed enzyme in step S4 consists of β-glucosidase, cellulase and pectinase in a mass ratio of 2:7:2.

[0110] The solvent 2 in step S4 is composed of water, betaine and 1,3-propylene glycol in a mass ratio of 8:1:2.5.

[0111] The preparation method of the solvent 2 in step S4 is as follows: betaine and 1,3-propylene glycol are stirred at 90° C. and 500 rpm for 10 min, and water is added to obtain the solvent 2.

[0112] The mass ratio of the medicinal residue 3, the mixed enzyme and the solvent 2 in step S4 is 1:0.08:8.

[0113] Preferably, in step S4, the mass ratio of the medicinal residue 3, the mixed enzyme and the solvent 2 is 1:0.06:6.

[0114] The temperature of the ultrasonic extraction in step S4 is 30° C., and the time of the ultrasonic extraction is 1 hour.

[0115] Example 3

[0116] The Chinese herbal medicine extract with the effect of dispelling cold has the same raw material composition as that in Example 1.

[0117] The preparation method of the Chinese herbal extract with the effect of dispelling cold is as follows:

[0118] Step S1, crushing cinnamon twig and ginger, soaking them in water, heating them under reflux, filtering them, and obtaining volatile oil, filtrate and medicinal residue 1;

[0119] Step S2, mixing the filtrate, peony root, liquorice, ephedra and ethanol aqueous solution, heating to reflux, filtering to obtain extract 2 and residue 2, and concentrating the extract 2 to obtain extract 1;

[0120] Step S3, mixing Atractylodes macrocephala, Rhizoma Anemarrhenae, Radix Saposhnikoviae, Radix Aconiti Lateralis Preparata, dregs 1, dregs 2 and solvent 1, heating and refluxing, filtering to obtain extract 2 and dregs 3, and concentrating extract 2 to obtain extract 2;

[0121] Step S4, mixing the drug residue 3, the mixed enzyme and the solvent 2, performing ultrasonic extraction, filtering and obtaining an extract 3, and concentrating the extract 3 to obtain an extract 3;

[0122] Step S5, the volatile oil, extract 1, extract 2 and extract 3 are mixed evenly to obtain the Chinese medicine extract.

[0123] The amount of water added in step S1 is 8 times the total mass of cassia twig and ginger.

[0124] The soaking temperature in step S1 is 25° C. and the soaking time is 2 h.

[0125] The heating reflux temperature in step S1 is 90° C., and the heating reflux time is 4 h.

[0126] The mass concentration of the ethanol aqueous solution in step S2 is 75%.

[0127] The amount of ethanol aqueous solution added in step S2 is 6 times the total mass of peony, liquorice and ephedra.

[0128] The heating reflux temperature in step S2 is 80° C. and the time is 4 hours.

[0129] The concentration in step S2 specifically includes: concentrating to twice the amount of extract 2.

[0130] The solvent 1 in step S3 is composed of water, caprylic acid capric acid macrogol glyceride and castor oil polyoxyethylene ether in a mass ratio of 30:3:1.

[0131] The heating reflux temperature in step S3 is 85° C. and the time is 3 h.

[0132] The amount of solvent 1 added in step S3 is 8 times the total mass of Atractylodes macrocephala, Rhizoma Anemarrhenae, Radix Saposhnikoviae, Radix Aconitum, dregs 1 and dregs 2.

[0133] The concentration in step S3 specifically includes: concentrating to twice the volume of extract 3.

[0134] The mixed enzyme in step S4 consists of β-glucosidase, cellulase and pectinase in a mass ratio of 1.5:5:1.

[0135] The solvent 2 in step S4 is composed of water, betaine and 1,3-propylene glycol in a mass ratio of 6:1:2.

[0136] The preparation method of the solvent 2 in step S4 is as follows: betaine and 1,3-propylene glycol are stirred at 80° C. and 400 rpm for 12 min, and water is added to obtain the solvent 2.

[0137] The mass ratio of the medicinal residue 3, the mixed enzyme and the solvent 2 in step S4 is 1:0.06:6.

[0138] The temperature of the ultrasonic extraction in step S4 is 25° C., and the time of the ultrasonic extraction is 2 h.

[0139] Comparative Example 1

[0140] The difference from Example 1 is that the solvent 1 in step S3 is composed of water, stearamidopropyl hydroxypropyl sulfobetaine and castor oil polyoxyethylene ether in a mass ratio of 30:3:1; the rest are the same.

[0141] Comparative Example 2

[0142] The difference from Example 1 is that the solvent 1 in step S3 is composed of water, caprylic acid capric acid macrogol glyceride and palm kernel oil amidopropyl betaine in a mass ratio of 30:3:1; the rest are the same.

[0143] Comparative Example 3

[0144] The difference from Example 1 is that the solvent 1 in step S3 is composed of water, betaine and 1,3-propylene glycol in a mass ratio of 6:1:2; the rest are the same.

[0145] Comparative Example 4

[0146] The difference from Example 1 is that the solvent 2 in step S4 is composed of water, choline chloride and 1,3-propylene glycol in a mass ratio of 6:1:2; the rest are the same.

[0147] Comparative Example 5

[0148] The difference from Example 1 is that the solvent 2 in step S4 is composed of water, caprylic acid capric acid polyethylene glycol glyceride and castor oil polyoxyethylene ether in a mass ratio of 30:3:1; the rest are the same.

[0149] Comparative Example 6

[0150] The difference from Example 1 is that the mixed enzyme in step S4 is composed of β-glucosidase, cellulase and neutral protease in a mass ratio of 1.5:5:1; the rest are the same.

[0151] Comparative Example 7

[0152] The difference from Example 1 is that the mixed enzyme in step S4 is composed of xylanase, cellulase and pectinase in a mass ratio of 1.5:5:1; the rest are the same.

[0153] Comparative Example 8

[0154] The difference from Example 3 is that the mixed enzyme in step S4 is composed of cellulase and pectinase in a mass ratio of 6.5:1; the rest are the same.

[0155] Comparative Example 9

[0156] The difference from Example 1 is that the preparation method of the Chinese herbal extract with the effect of dispelling cold is:

[0157] Step S1, mixing cinnamon twig, ginger, peony root, liquorice, ephedra and ethanol aqueous solution, heating to reflux, filtering to obtain extract 1 and medicinal residue 1, and concentrating the extract 1 to obtain extract 1;

[0158] Step S2, mixing Atractylodes macrocephala, Rhizoma Anemarrhenae, Radix Saposhnikoviae, Radix Aconiti Lateralis Preparata, medicinal residue 1 and solvent 1, heating and refluxing, filtering to obtain extract 2 and medicinal residue 2, and concentrating the extract 2 to obtain extract 2;

[0159] Step S3, mixing the drug residue 2, the mixed enzyme and the solvent 2, performing ultrasonic extraction, filtering and obtaining an extract 3, and concentrating the extract 3 to obtain an extract 3;

[0160] Step S4, evenly mix extract 1, extract 2 and extract 3 to obtain a traditional Chinese medicine extract.

[0161] The mass concentration of the ethanol aqueous solution in step S1 is 75%.

[0162] The amount of ethanol aqueous solution added in step S1 is 6 times the total mass of cinnamon twig, ginger, peony root, liquorice and ephedra.

[0163] The heating reflux temperature in step S1 is 80° C. and the time is 4 hours.

[0164] The concentration in step S1 specifically includes: concentrating to twice the amount of extract 2.

[0165] The solvent 1 in step S2 is composed of water, caprylic acid capric acid macrogol glyceride and castor oil polyoxyethylene ether in a mass ratio of 30:3:1.

[0166] The heating reflux temperature in step S2 is 85° C. and the time is 3 h.

[0167] The amount of solvent 1 added in step S2 is 8 times the total mass of Atractylodes macrocephala, Rhizoma Anemarrhenae, Radix Saposhnikoviae, Radix Aconitum, dregs 1 and dregs 2.

[0168] The concentration in step S2 specifically includes: concentrating to twice the amount of the extract 3.

[0169] The mixed enzyme in step S3 consists of β-glucosidase, cellulase and pectinase in a mass ratio of 1.5:5:1.

[0170] The solvent 2 in step S3 is composed of water, betaine and 1,3-propylene glycol in a mass ratio of 6:1:2.

[0171] The preparation method of the solvent 2 in step S3 is as follows: betaine and 1,3-propylene glycol are stirred at 80° C. and 400 rpm for 12 min, and water is added to obtain the solvent 2.

[0172] The mass ratio of the medicinal residue 3, the mixed enzyme and the solvent 2 in step S3 is 1:0.06:6.

[0173] The temperature of the ultrasonic extraction in step S3 is 25° C., and the time of the ultrasonic extraction is 2 h.

[0174] Effect Experiment

[0175] 1. Construction of a rat model of rheumatoid arthritis: 150 SPF male SD rats, 6-8 weeks old, weighing 180-220g; randomly divided into 15 groups: a normal control group, a model control group, Example 1, Example 2, Example 3, Comparative Example 1, Comparative Example 2, Comparative Example 3, Comparative Example 4, Comparative Example 5, Comparative Example 6, Comparative Example 7, Comparative Example 8, Comparative Example 9 and a Guizhi Shaoyao Zhimu Decoction control group; 10 rats in each group; except the normal control group, bovine type II collagen was used to induce the construction of a rat model of rheumatoid arthritis, the method being: on the first day of the experiment, the emulsion of bovine type II collagen and incomplete Freund's adjuvant mixed in equal amounts was intradermally injected at multiple points at the base of the rat's tail, 0.2 mL / rat, and the rats were re-immunized on the 8th day (0.1 mL / rat), and the modeling lasted for a total of 14 days to obtain a rat model of rheumatoid arthritis, and the normal group was injected with an equal amount of physiological saline.

[0176] 2. Administration: Administration began on day 15. The corresponding Chinese herbal extracts of 4.5 g / kg were administered by gavage to the examples and comparative examples, respectively. The normal control group and the model control group were given an equal amount of distilled water. The Guizhi Shaoyao Zhimu Decoction control group was given a dose of 4.5 g / kg, once a day, for 14 consecutive days.

[0177] The preparation method of Guizhi Shaoyao Zhimu Decoction is as follows: take 12g of cinnamon twig, 9g of white peony root, 12g of Anemarrhena, 12g of Saposhnikovia divaricata, 15g of Atractylodes macrocephala, 6g of Ephedra, 6g of Licorice, 15g of ginger, and 6g of Paofuzi, add 16 times the amount of water, reflux extraction at 98°C for 3 times, each time for 1 hour, combine the filtrate, and concentrate the filtrate to obtain a concentrated solution, and the density of the concentrated solution at 25°C is 1.087g / mL.

[0178] 3. Detection:

[0179] 3.1. After administration, the paw volume swelling meter was used to measure the paw swelling of rats in each group.

[0180] The formula for calculating the degree of plantar swelling is: (toe volume after inflammation - toe volume before inflammation) / toe volume before inflammation.

[0181] 3.2. After administration, the primary lesions on the inflamed side of each group of rats were observed, and the arthritis lesion index was calculated. The arthritis lesion index was evaluated using a 5-grade scoring method, with no redness or swelling being assigned a score of 0, redness or swelling of the little toe joint being assigned a score of 1, swelling of the toe joint and the plantar being assigned a score of 2, swelling of the paw below the ankle being assigned a score of 3, and swelling of the entire paw including the ankle being assigned a score of 4.

[0182] 3.3. After administration, the rats were anesthetized, blood was collected from the abdominal aorta, and serum was collected to test the levels of TNF-α and IL-1β according to the instructions of the kit.

[0183] 4. SPSS26.0 statistical software was used for data analysis. The results were expressed as x±s. One-way analysis of variance was used for comparison among multiple groups. Non-parametric tests were used when the data did not conform to normal distribution. P < 0.05 was considered statistically significant.

[0184] 5. Results Analysis

[0185] 5.1 The results of plantar swelling and arthritis index are shown in Table 1.

[0186] Table 1

[0187]

[0188] Note: Compared with the normal control group, ** P<0.01; compared with the model control group, ## P<0.01, # P<0.05. 5.2. TNF-α, IL-1β levels: The results are shown in Table 2.

[0189] Table 2

[0190]

[0191]

[0192] Note: Compared with the normal control group, * P<0.05; compared with the model control group, # P<0.05.

[0193] As can be seen from Table 1 and Table 2, when the dosage is 4.5 g / kg, after 14 days of administration: compared with the model control group, the Chinese medicine extracts of Examples 1-3 have extremely significant differences in plantar swelling and arthritis lesion index, and significant differences in TNF-α and IL-1β levels; while there is no significant difference in the Guizhi Shaoyao Zhimu Decoction group, indicating that the Chinese medicine extracts of the present invention have excellent effects in treating rheumatoid arthritis;

[0194] Comparative Example 1 and Comparative Example 2 replaced a certain component in Solvent 1, resulting in significant differences in the paw swelling and arthritis lesion index of the obtained Chinese medicine extracts compared with the model control group, but the effect of treating rheumatoid arthritis decreased, resulting in increased paw swelling and arthritis lesion index of rats; while there was no significant difference in TNF-α and IL-1β levels;

[0195] In Comparative Example 3, since solvent 1 was replaced by solvent 2, the obtained Chinese medicine extract had no significant difference in plantar swelling, arthritis lesion index, TNF-α, and IL-1β levels compared with the model control group;

[0196] Comparative Example 4 Due to the replacement of a component in solvent 2, the obtained Chinese medicine extract showed significant differences in the swelling degree of the paw and the arthritis lesion index compared with the model control group, but the effect of treating rheumatoid arthritis decreased, resulting in an increase in the swelling degree of the paw and the arthritis lesion index of the rats; while there was no significant difference in the levels of TNF-α and IL-1β;

[0197] In Comparative Example 5, since solvent 2 was replaced by solvent 1, the obtained Chinese medicine extract had no significant difference in plantar swelling, arthritis lesion index, TNF-α, and IL-1β levels compared with the model control group;

[0198] Comparative Examples 6 and 7 replaced a certain component in the mixed enzyme, resulting in significant differences in the paw swelling and arthritis lesion index of the obtained Chinese medicine extracts compared with the model control group, but the effect of treating rheumatoid arthritis decreased, resulting in increased paw swelling and arthritis lesion index of rats; while there was no significant difference in TNF-α and IL-1β levels;

[0199] In Comparative Example 8, since β-glucosidase was replaced with cellulase of equal mass, the obtained Chinese medicine extract had no significant difference in plantar swelling, arthritis lesion index, TNF-α, and IL-1β levels compared with the model control group;

[0200] In Comparative Example 9, since the Chinese medicinal materials were not extracted in batches, the obtained Chinese medicinal extracts showed significant differences in plantar swelling and arthritis lesion index compared with the model control group, but the effect on the treatment of rheumatoid arthritis was reduced, resulting in increased plantar swelling and arthritis lesion index of rats; while there were no significant differences in TNF-α and IL-1β levels.

[0201] The above is a further description of the present invention in conjunction with specific embodiments, but these embodiments are merely exemplary and do not constitute any limitation on the scope of the present invention. It should be understood by those skilled in the art that the details and forms of the technical solution of the present invention may be modified or replaced without departing from the spirit and scope of the present invention, but these modifications and replacements all fall within the scope of protection of the present invention.

Claims

1. A method for preparing a Chinese herbal medicine extract having a cold-dispelling effect, characterized in that: The following steps are involved: Step S1, crushing cinnamon twig and ginger, soaking them in water, heating them under reflux, filtering them, and obtaining volatile oil, filtrate and medicinal residue 1; Step S2, mixing the filtrate, peony, liquorice, ephedra and ethanol aqueous solution, heating to reflux, filtering to obtain extract 2 and residue 2, and concentrating the extract 2 to obtain extract 1; Step S3, mixing Atractylodes macrocephala, Rhizoma Anemarrhenae, Radix Saposhnikoviae, Radix Aconiti Lateralis Preparata, dregs 1, dregs 2 and solvent 1, heating and refluxing, filtering to obtain extract 2 and dregs 3, and concentrating extract 2 to obtain extract 2; Step S4, mixing the drug residue 3, the mixed enzyme and the solvent 2, performing ultrasonic extraction, filtering and obtaining an extract 3, and concentrating the extract 3 to obtain an extract 3; Step S5, mixing the volatile oil, extract 1, extract 2 and extract 3 evenly to obtain a Chinese medicine extract; The solvent 1 in step S3 includes water, caprylic acid capric acid macrogol glyceride and castor oil polyoxyethylene ether; The solvent 2 in step S4 includes water, betaine and 1,3-propylene glycol.

2. The preparation method according to claim 1, characterized in that: The raw materials of the traditional Chinese medicine extract include, by weight: 12 parts of cinnamon twig, 9 parts of peony root, 6 parts of liquorice, 12 parts of ephedra, 15 parts of ginger, 15 parts of atractylodes macrocephala, 12 parts of anemarrhena, 12 parts of siler, and 10 parts of processed aconite.

3. The preparation method according to claim 1 or 2, characterized in that: The amount of water added in step S1 is 5-10 times the total mass of cassia twig and ginger; the soaking temperature in step S1 is 25-30° C.; the heating reflux temperature in step S1 is 85-95° C., and the heating reflux time is 3-7 hours.

4. The preparation method according to claim 1 or 2, characterized in that: The amount of ethanol aqueous solution added in step S2 is 4-8 times the total mass of peony, liquorice and ephedra; the temperature of heating reflux in step S2 is 75-85°C and the time is 3-5h.

5. The preparation method according to claim 1 or 2, characterized in that: The solvent 1 in step S3 comprises water, caprylic acid capric acid macrogol glyceride and castor oil polyoxyethylene ether in a mass ratio of 25-35:2-5:

1.

6. The preparation method according to claim 5, characterized in that: The solvent 1 in step S3 comprises water, caprylic acid capric acid macrogol glyceride and castor oil polyoxyethylene ether in a mass ratio of 30:3:

1.

7. The preparation method according to claim 1 or 2, characterized in that: The amount of solvent 1 added in step S3 is 5-10 times the total mass of Atractylodes macrocephala, Anemarrhena asphodeloides, Saposhnikovia divaricata, Aconitum carmichaelii, dregs 1 and dregs 2; the temperature of heating reflux in step S3 is 80-90° C., and the time is 2-5 h.

8. The preparation method according to claim 1 or 2, characterized in that: The mixed enzyme in step S4 includes β-glucosidase, cellulase and pectinase in a mass ratio of 1-2:3-7:1-2.

9. The preparation method according to claim 8, characterized in that: The mixed enzyme in step S4 includes β-glucosidase, cellulase and pectinase in a mass ratio of 1.5:5:

1.

10. The preparation method according to claim 1 or 2, characterized in that: The solvent 2 in step S4 comprises water, betaine and 1,3-propylene glycol in a mass ratio of 4-8:1:1.5-2.

5.

11. The preparation method according to claim 10, characterized in that: The solvent 2 in step S4 includes water, betaine and 1,3-propylene glycol in a mass ratio of 6:1:

2.

12. The preparation method according to claim 11, characterized in that: The preparation method of the solvent 2 in step S4 is as follows: betaine and 1,3-propylene glycol are stirred at 70-90° C. and 300-500 rpm for 10-15 min, and water is added to obtain the solvent 2.

13. The preparation method according to claim 1 or 2, characterized in that: The mass ratio of the medicinal residue 3, the mixed enzyme and the solvent 2 in step S4 is 1:0.04-0.08:4-8; the temperature of the ultrasonic extraction in step S4 is 25-30°C, and the time of the ultrasonic extraction is 1-3h.

14. The preparation method according to claim 13, characterized in that: The mass ratio of the medicinal residue 3, the mixed enzyme and the solvent 2 in step S4 is 1:0.06:6; the temperature of the ultrasonic extraction in step S4 is 25°C, and the time of the ultrasonic extraction is 2h.

15. A Chinese medicinal extract with cold-dispelling effect obtained by the preparation method according to any one of claims 1 to 14.

16. Use of the Chinese medicinal extract with the effect of expelling cold obtained by the preparation method according to any one of claims 1 to 14 in the preparation of drugs for rheumatoid arthritis.

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