Traditional Chinese medicine extract with cold dispelling effect, preparation method thereof and application thereof in preparing medicine for rheumatoid arthritis

Abstract

The application belongs to the technical field of traditional Chinese medicine, and particularly relates to a traditional Chinese medicine extract with cold dispelling effect, a preparation method thereof and application of the traditional Chinese medicine extract in preparation of a rheumatoid arthritis medicine. The preparation method comprises the following steps: heating and refluxing guizhi and ginger with water, filtering, obtaining volatile oil, a filtrate and dregs 1; heating and refluxing the filtrate, peony, licorice, ephedra and an ethanol aqueous solution, obtaining extract 2 and dregs 2, and concentrating the extract 2 to obtain extractum 1; heating and refluxing baizhu, zhimu, fangfeng, fried aconite root, the dregs 1, the dregs 2 and solvent 1, obtaining extract 2 and dregs 3, and concentrating the extract 2 to obtain extractum 2; ultrasonic extracting the dregs 3, mixed enzymes and solvent 2, obtaining extract 3, and concentrating the extract 3 to obtain extractum 3; uniformly mixing the volatile oil, the extractum 1, the extractum 2 and the extractum 3 to obtain the traditional Chinese medicine extract; compared with other extraction methods, the traditional Chinese medicine extract prepared by the method has a remarkable cold dispelling effect and rheumatoid arthritis treatment effect.

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 a cold-dispelling effect, a preparation method thereof, and application of the extract in preparing a rheumatoid arthritis medicine. Background Art

[0002] Guizhi Shaoyao Zhimu Decoction is a prescription from the Synopsis of the Golden Chamber, written by Zhang Zhongjing, the Han Dynasty medical sage. It consists of cinnamon twig, peony root, licorice root, ephedra, ginger, white atractylodes, anemarrhena, saposhnikovia root, and aconite root. It is primarily used to treat joint pain, weakness, swollen feet, dizziness, shortness of breath, fever and nausea, a reddish tongue with a white coating, and a weak and rapid pulse. It is primarily used to treat rheumatoid arthritis, rheumatic arthritis, polymyalgia rheumatica, fibromyalgia, sciatica, femoral head necrosis, and tenosynovitis. It is also used for measles-related pneumonia, tracheitis, cor pulmonale with heart failure, and deep tissue inflammation.

[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] The Guizhi Shaoyao Zhimu decoction is improved and prepared 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 formula slices of the Guizhi Shaoyao Zhimu prescription preparation, crushing them, and mixing them; 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. This technical solution uses the paste yield, cinnamaldehyde in cinnamon twig, paeoniflorin in white peony, mangiferin in Zhimu, glycyrrhizic acid in licorice, and inflammatory factor IL-6 as 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. The prepared extract has a high content of active ingredients and a significant anti-inflammatory effect, and can be used for mass production.

[0005] Zeng Hairong et al. also published "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 result in 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 traditional Chinese medicine extract with a cold-dispelling effect, a preparation method thereof, and an application of the extract in preparing a rheumatoid arthritis drug.

[0008] To achieve the above object, 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 under reflux, and filtering to obtain volatile oil, filtrate and medicinal residue 1;

[0011] Step S2, mixing the filtrate, peony root, liquorice, ephedra and ethanol aqueous solution, heating under reflux, filtering to obtain extract 2 and medicinal 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, Medicinal Residue 1, Medicinal Residue 2 and Solvent 1, heating and refluxing, filtering to obtain Extract 3 and Medicinal Residue 3, and concentrating Extract 3 to obtain Extract 2;

[0013] Step S4, mixing the medicinal residue 3, the mixed enzyme, and the solvent 2, performing ultrasonic extraction, filtering, and obtaining an extract 4, and concentrating the extract 4 to obtain an extract 3;

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

[0015] The solvent 1 in step S3 comprises 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 Chinese medicine extract include, by mass: 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 root.

[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 hours.

[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 temperature of the heating reflux 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 root, liquorice 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 / capric 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 temperature of the heating reflux in step S3 is 85° C. and the time is 3 hours.

[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 Aconiti Lateralis Preparata, dregs 1 and dregs 2.

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

[0034] In some preferred embodiments, the enzyme mixture 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 extract with a cold-dispelling effect obtained by the above preparation method.

[0045] The third aspect of the present invention provides the use of the Chinese herbal 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 rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

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

[0050] Unless otherwise indicated, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein may also be used in the practice 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 any conflict with any incorporated document, the contents 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 described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from the description of the invention. The description and examples are intended to be illustrative 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 macrogol glycerides were purchased from Gattefosse (Shanghai) Trading Co., Ltd. under the trade name LABRASOL® 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 white atractylodes, 12 parts of anemarrhena, 12 parts of siler, and 10 parts of processed aconite root.

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

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

[0070] Step S2, mixing the filtrate, peony root, liquorice, ephedra and ethanol aqueous solution, heating under reflux, filtering to obtain extract 2 and medicinal 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, Medicinal Residues 1, Medicinal Residues 2 and Solvent 1, heating and refluxing, filtering to obtain Extract 3 and Medicinal Residues 3, and concentrating Extract 2 to obtain Extract 2;

[0072] Step S4, mixing the medicinal residue 3, the mixed enzyme, and the solvent 2, performing ultrasonic extraction, filtering, and obtaining an extract 4, and concentrating the extract 4 to obtain an extract 3;

[0073] Step S5: Evenly mix the volatile oil, extract 1, extract 2, and extract 3 to obtain a Chinese medicine extract.

[0074] The amount of water added in step S1 is 5 times the total mass of Ramulus Cinnamomi and Rhizoma Zingiberis Recens.

[0075] The temperature of the soaking in step S1 is 25℃, and the time of the soaking is 1h.

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

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

[0078] The amount of the aqueous ethanol solution added in step S2 is 4 times the total mass of Paeoniae Radix Alba, Glycyrrhizae Radix et Rhizoma, and Ephedrae Herba.

[0079] The temperature of the heating reflux in step S2 is 75℃, and the time is 5h.

[0080] The concentration in step S2 is specifically: concentrated to 2 times the extraction liquid 2.

[0081] The solvent 1 in step S3 is composed of water, caprylocaproyl macrogolglycerides, and castor oil polyoxyl ether with a mass ratio of 25:2:1.

[0082] The temperature of the heating reflux in step S3 is 80℃, and the time is 5h.

[0083] The amount of the solvent 1 added in step S3 is 5 times the total mass of Atractylodis Macrocephalae Rhizoma, Anemarrhenae Rhizoma, Saposhnikoviae Radix, Radix Aconiti Praeparata, dregs 1, and dregs 2.

[0084] The concentration in step S3 is specifically: concentrated to 2 times the extraction liquid 3.

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

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

[0087] The preparation method of the solvent 2 in step S4 is: betaine and 1,3-propanediol are stirred at 70℃ at a speed of 300rpm for 15min, and then water is added to obtain the solvent 2.

[0088] The mass ratio of the dregs 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℃, and the time of the ultrasonic extraction is 3h.

[0090] Example 2

[0091] The Chinese herbal 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 medicinal extract with the cold-dispelling effect is as follows:

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

[0094] Step S2, mixing the filtrate, peony root, liquorice, ephedra and ethanol aqueous solution, heating under reflux, filtering to obtain extract 2 and medicinal 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, Medicinal Residue 1, Medicinal Residue 2 and Solvent 1, heating and refluxing, filtering to obtain Extract 3 and Medicinal Residue 3, and concentrating Extract 3 to obtain Extract 2;

[0096] Step S4, mixing the medicinal residue 3, the mixed enzyme, and the solvent 2, performing ultrasonic extraction, filtering, and obtaining an extract 4, and concentrating the extract 4 to obtain an extract 3;

[0097] Step S5: Evenly mix the volatile oil, extract 1, extract 2, and extract 3 to obtain a 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 root, 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 is specifically: 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, Anemarrhena asphodeloides, Saposhnikovia divaricata, Aconitum carmichaelii, dregs 1 and dregs 2.

[0108] The concentration in step S3 is specifically concentrated to 3 times of the extraction liquid 3.

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

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

[0111] The preparation method of the solvent 2 in step S4 is that betaine and 1,3-propanediol are stirred at 90℃ for 10 min at a speed of 500 rpm, and then water is added to obtain the solvent 2.

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

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

[0114] The temperature of the ultrasonic extraction in step S4 is 30℃, and the time of the ultrasonic extraction is 1 h.

[0115] Example 3

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

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

[0118] Step S1, grinding cassia twig and raw ginger, soaking in water, heating reflux, filtering to obtain volatile oil, filtrate and dregs 1;

[0119] Step S2, mixing the filtrate, peony, licorice, ephedra and ethanol aqueous solution, heating reflux, filtering to obtain extraction liquid 2 and dregs 2, concentrating the extraction liquid 2 to obtain extract 1;

[0120] Step S3, mixing white atractylodes rhizome, anemarrhena asphodeloides, sileris, prepared aconite root, dregs 1, dregs 2 and solvent 1, heating reflux, filtering to obtain extraction liquid 3 and dregs 3, concentrating the extraction liquid 3 to obtain extract 2;

[0121] Step S4, mixing the dregs 3, mixed enzyme and solvent 2, ultrasonic extraction, filtering to obtain extraction liquid 4, concentrating the extraction liquid 4 to obtain extract 3;

[0122] Step S5, uniformly mixing the volatile oil, extract 1, extract 2 and extract 3 to obtain the traditional Chinese medicine extract.

[0123] The added amount of water in step S1 is 8 times of the total mass of cassia twig and raw 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 root, liquorice and ephedra.

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

[0129] The concentration in step S2 is specifically: concentrating to twice the volume 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, Anemarrhena asphodeloides, Saposhnikovia divaricata, Aconitum carmichaelii, dregs 1 and dregs 2.

[0133] The concentration in step S3 is specifically: 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 minutes, 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 macrogol 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 consists 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 enzyme mixture in step S4 consists 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 consists 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 cold-dispelling effect is:

[0157] Step S1, mixing cinnamon twig, ginger, peony root, liquorice, ephedra and ethanol aqueous solution, heating under 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 under reflux, filtering to obtain extract 2 and medicinal residue 2, and concentrating the extract 2 to obtain extract 2;

[0159] Step S3, mixing the medicinal 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 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 h.

[0164] The concentration in step S1 is specifically: concentrating to twice the volume 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, Anemarrhena asphodeloides, Saposhnikovia divaricata, Aconitum carmichaelii, dregs 1 and dregs 2.

[0168] The concentration in step S2 specifically includes: concentrating to twice the volume 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 minutes, 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. Constructing rheumatoid arthritis rat model: 150 SPF male SD rats, 6-8 weeks old, weighing 180-220 g; randomly divided into 15 groups: normal control group, 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 Guizhi Shaoyao Zhimu decoction control group; 10 rats in each group; except the normal control group, all were induced to construct rheumatoid arthritis rat model with bovine collagen type II, the method was as follows: on the first day of the experiment, the rats were injected with emulsion of equal amount of bovine collagen type II and incomplete Freund's adjuvant at the root of the tail, 0.2 mL per rat, on the 8th day, the rats were immunized again (0.1 mL per rat), a total of 14 days of modeling, rheumatoid arthritis rat model was obtained, the normal group was injected with equal amount of normal saline.

[0176] 2. Drug administration: 15 days of drug administration, examples and comparative examples were given corresponding traditional Chinese medicine extract 4.5 g / kg by gavage, the normal control group and the model control group were given equal amount of distilled water, the Guizhi Shaoyao Zhimu decoction control group was given 4.5 g / kg, once a day, for 14 days.

[0177] The preparation method of Guizhi Shaoyao Zhimu decoction was as follows: 12 g of cassia twig, 9 g of white peony root, 12 g of anemarrhena, 12 g of wind-protecting, 15 g of atractylodes, 6 g of ephedra, 6 g of licorice, 15 g of ginger, 6 g of processed aconite were taken, 16 times the amount of water was added, reflux extraction was carried out for 3 times at 98℃, 1 h each time, the filtrates were combined, the obtained filtrate was concentrated to obtain concentrated liquid, the density of the obtained concentrated liquid was 1.087 g / mL at 25℃.

[0178] 3. Detection:

[0179] 3.1, after drug administration, the foot volume swelling meter was used to measure the foot swelling degree of rats in each group.

[0180] The formula for calculating the foot swelling degree was: (inflamed toe volume-inflamed toe volume before inflammation) / inflamed toe volume before inflammation.

[0181] 3.2, after drug administration, the primary lesion of the inflamed side of rats in each group was observed, and the arthritis lesion index was calculated,

[0182] The arthritis lesion index was evaluated according to the 5-grade scoring method, no redness and swelling was 0 point, redness and swelling of the toe joint was 1 point, swelling of the toe joint and foot was 2 points, swelling of the foot below the ankle joint was 3 points, and swelling of the whole foot including the ankle joint was 4 points.

[0183] 3.3, after drug administration, the rats were anesthetized, blood was taken from the abdominal aorta, serum was taken, and the levels of TNF-α and IL-1β were tested according to the instructions of the kit.

[0184] 4. SPSS 26.0 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 results did not conform to the normal distribution. P < 0.05 was considered statistically significant.

[0185] 5. Results Analysis

[0186] 5.1 The results of plantar swelling and arthritis index are shown in Table 1.

[0187] Table 1

[0188]

[0189] Note: Compared with the normal control group, ** P<0.01; compared with the model control group, ## P < 0.01, # P<0.05.

[0190] 5.2. TNF-α and IL-1β levels: The results are shown in Table 2.

[0191] Table 2

[0192]

[0193] Note: Compared with the normal control group, * P<0.05; compared with the model control group, # P<0.05.

[0194] As can be seen from Tables 1 and 2, when the dosage was 4.5 g / kg, 14 days after administration, the Chinese herbal extracts of Examples 1-3 showed extremely significant differences in plantar swelling and arthritis index compared with the model control group, and significant differences in TNF-α and IL-1β levels; while there was no significant difference in the Guizhi Shaoyao Zhimu Decoction group, indicating that the Chinese herbal extracts of the present invention have excellent therapeutic effects on rheumatoid arthritis.

[0195] Comparative Examples 1 and 2, when a component in Solvent 1 was replaced, resulted in the obtained Chinese herbal extracts showing significant differences in paw swelling and arthritis lesion index compared to the model control group, but decreased efficacy in treating rheumatoid arthritis, resulting in increased paw swelling and arthritis lesion index in rats; however, there were no significant differences in TNF-α and IL-1β levels.

[0196] In Comparative Example 3, since solvent 1 was replaced by solvent 2, the obtained Chinese medicine extract showed no significant differences in plantar swelling, arthritis index, TNF-α, and IL-1β levels compared with the model control group;

[0197] The comparative example 4, due to replacing a component in the solvent 2, the obtained traditional Chinese medicine extract has significant difference in the foot swelling degree and the arthritis lesion index compared with the model control group, but the effect of treating rheumatoid arthritis decreases, leading to the increase of the foot swelling degree and the arthritis lesion index of the rats; and the TNF-α and IL-1β levels have no significant difference.

[0198] The comparative example 5, due to replacing the solvent 2 with the solvent 1, the obtained traditional Chinese medicine extract has no significant difference in the foot swelling degree, the arthritis lesion index, the TNF-α and IL-1β levels compared with the model control group.

[0199] The comparative example 6 and the comparative example 7, due to replacing a component in the mixed enzyme, the obtained traditional Chinese medicine extract has significant difference in the foot swelling degree and the arthritis lesion index compared with the model control group, but the effect of treating rheumatoid arthritis decreases, leading to the increase of the foot swelling degree and the arthritis lesion index of the rats; and the TNF-α and IL-1β levels have no significant difference.

[0200] The comparative example 8, due to replacing the β-glucosidase with cellulase of the same quality, the obtained traditional Chinese medicine extract has no significant difference in the foot swelling degree, the arthritis lesion index, the TNF-α and IL-1β levels compared with the model control group.

[0201] The comparative example 9, due to not carrying out batch extraction on the traditional Chinese medicinal materials, the obtained traditional Chinese medicine extract has significant difference in the foot swelling degree and the arthritis lesion index compared with the model control group, but the effect of treating rheumatoid arthritis decreases, leading to the increase of the foot swelling degree and the arthritis lesion index of the rats; and the TNF-α and IL-1β levels have no significant difference.

[0202] The above is the further description of the present application combined with the specific embodiments, but these embodiments are only exemplary and do not constitute any limitation on the scope of the present application. Those skilled in the art should understand that the details and forms of the technical solutions of the present application can be modified or replaced without departing from the spirit and scope of the present application, and these modifications and replacements all fall within the protection scope of the present application.

Claims

1. A method for preparing a Chinese herbal 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 under reflux, and filtering to obtain volatile oil, filtrate and medicinal residue 1; Step S2, mixing the filtrate, peony root, liquorice, ephedra and ethanol aqueous solution, heating under reflux, filtering to obtain extract 2 and medicinal residue 2, and concentrating the extract 2 to obtain extract 1; Step S3, mixing Atractylodes macrocephala, Rhizoma Anemarrhenae, Radix Saposhnikoviae, Radix Aconiti Lateralis Preparata, Medicinal Residues 1, Medicinal Residues 2 and Solvent 1, heating and refluxing, filtering to obtain Extract 3 and Medicinal Residues 3, and concentrating Extract 3 to obtain Extract 2; Step S4, mixing the medicinal residue 3, the mixed enzyme, and the solvent 2, performing ultrasonic extraction, filtering, and obtaining an extract 4, and concentrating the extract 4 to obtain an extract 3; Step S5, uniformly mixing the volatile oil, extract 1, extract 2 and extract 3 to obtain a Chinese medicine extract; The solvent 1 in step S3 is water, caprylic acid capric acid polyethylene glycol glyceride and castor oil polyoxyethylene ether; The solvent 2 in step S4 is water, betaine and 1,3-propylene glycol; The raw materials of the Chinese herbal medicine extract are calculated by mass as follows: 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, 12 parts of saposhnikovia divaricata, and 10 parts of processed aconite root; The mixed enzyme in step S4 is β-glucosidase, cellulase and pectinase in a mass ratio of 1-2:3-7:1-2.

2. The preparation method according to claim 1, 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.

3. The preparation method according to claim 1, 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 heating reflux temperature in step S2 is 75-85° C. and the time is 3-5 hours.

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

1.

5. The preparation method according to claim 4, characterized in that The solvent 1 in step S3 is water, caprylic acid capric acid polyethylene glycol glyceride and castor oil polyoxyethylene ether in a mass ratio of 30:3:

1.

6. The preparation method according to claim 1, 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 hours.

7. The preparation method according to claim 1, characterized in that The mixed enzyme in step S4 is β-glucosidase, cellulase and pectinase in a mass ratio of 1.5:5:

1.

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

5.

9. The preparation method according to claim 8, characterized in that The solvent 2 in step S4 is water, betaine and 1,3-propylene glycol in a mass ratio of 6:1:

2.

10. The preparation method according to claim 9, 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 minutes, and water is added to obtain the solvent 2.

11. The preparation method according to claim 1, 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.

12. The preparation method according to claim 11, 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 2 h.

13. A Chinese medicinal extract with a cold-dispelling effect obtained by the preparation method according to any one of claims 1 to 12.

14. Use of the Chinese medicinal extract with the cold-dispelling effect obtained by the preparation recipe according to any one of claims 1 to 12 in the preparation of a drug for rheumatoid arthritis.

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