Traditional Chinese medicine composition as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a traditional Chinese medicine composition and a preparation method and application thereof. The traditional Chinese medicine composition is prepared from the following raw materials in parts by mass: 20 to 60 parts of rhizoma acori calami, 30 to 50 parts of radix aconiti lateralis, 20 to 60 parts of benzoin and 40 to 100 parts of catechu. The traditional Chinese medicine composition disclosed by the invention is simple and natural in component and clear in drug effect, can effectively reduce the contents of XOD, SUA, CRE, BUN, TNF-alpha, IL-6, D-Lac and DAO in serum of a gout mouse model, remarkably reduces the kidney index, and has a good drug effect on treating hyperuricemia and / or gout.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a traditional Chinese medicine composition and a preparation method and use thereof. Background Art

[0002] Hyperuricemia is a common metabolic disorder associated with a variety of serious diseases. Hyperuricemia is caused by excessive production of uric acid or insufficient excretion by the kidneys and intestines. It is not only a risk factor for gout, but is also closely related to the onset and progression of hypertension, diabetes, and kidney disease. Epidemiological studies have shown that the prevalence of hyperuricemia in China continues to rise, reaching 17.4%. At present, it is the second most common metabolic disorder after diabetes, and the patient population is gradually getting younger, which is expected to bring a significant burden to China's medical system in the next few years.

[0003] The general treatment strategy for hyperuricemia focuses on the use of xanthine oxidase inhibitors, such as allopurinol and febuxostat, to reduce uric acid levels, and the use of uricosuric agents, such as probenecid, benzbromarone, and sulfinpyrazone, to promote uric acid excretion. However, adverse reactions associated with hyperuricemia inhibitors and uricosuric agents, including diarrhea, headache, rash, severe allergies, and nephrotoxicity, often limit their use in the clinic. Therefore, it is crucial to find more effective and nontoxic natural products to treat hyperuricemia.

[0004] The incidence of hyperuricemia in Tibetan areas is significantly higher than in other parts of China, with a crude prevalence of up to 37.2%. Over the years, Tibetan medicine has accumulated a large number of drugs for the treatment of hyperuricemia, but traditional Tibetan medicine has complex ingredients, unclear mechanisms of action, and difficult to determine effective ingredients. Therefore, it is necessary to develop a Chinese medicine composition with simple ingredients, clear efficacy, and good therapeutic effects for the treatment of hyperuricemia and / or gout. Summary of the invention

[0005] The technical problem to be solved by the present invention is to provide a traditional Chinese medicine composition for treating hyperuricemia and / or gout with simple ingredients, clear efficacy and good therapeutic effect, which can significantly reduce the contents of XOD, SUA, CRE, BUN, TNF-α, IL-6, D-Lac and DAO in the serum of gout mouse model and reduce the renal index, and can effectively treat hyperuricemia and / or gout.

[0006] To achieve the above object, the present invention provides a Chinese medicine composition, which comprises the following ingredients by weight: 20-60 parts of calamus, 30-50 parts of iron hammer, 20-60 parts of benzoin, and 40-100 parts of catechu.

[0007] The calamus used in the present invention is the rhizome of the calamus plant Acorus calamus L. of the calamaceae family, has the effects of opening the mind, removing phlegm and regulating qi, and is a ministerial drug. It is commonly used for phlegm-induced coma, stroke, epilepsy, palpitations and forgetfulness, tinnitus and deafness, abdominal pain due to food accumulation, dysentery and diarrhea, rheumatic pain, eczema and scabies.

[0008] The iron club hammer adopted by the invention is the dried root tuber of the Ranunculaceae plant Aconitum pendulum Busch., which has the effects of dispelling wind and dampness, dispersing blood stasis and relieving pain, and is an adjuvant medicine for treating traumatic injury, bone fracture, rheumatic waist pain, frostbite and the like.

[0009] The benzoin used in the present invention is the dry resin of the white flower tree of the Styracaceae family (i.e. Vietnamese benzoin Styrax tonkinensis (Pierre) Craib ex Hart.). Indications: opening the mind and clearing the spirit, promoting qi and blood circulation, relieving pain, and serving as a guiding drug. It is used for stroke, phlegm syncope, qi stagnation, sudden syncope, coma caused by poisoning, heart and abdomen pain, postpartum blood fainting, and infantile convulsions.

[0010] The catechu used in the present invention is a decoction of peeled branches and stems of the leguminous plant Acacia catechu (Lf) Willci. It has the effects of promoting blood circulation and relieving pain, stopping bleeding and promoting tissue regeneration, absorbing dampness and astringing sores, clearing the lungs and resolving phlegm, and is a main drug. It is used for pain caused by falls, bleeding from trauma, vomiting blood, epistaxis, sores that do not astringe, eczema, wet sores, and cough due to lung heat.

[0011] The Chinese medicine composition including calamus, cyperus rotundus, benzoin and catechu provided by the invention is reasonably matched, and the four Chinese medicine components synergize to enhance the effect, and has a good therapeutic effect for treating hyperuricemia and / or gout.

[0012] Preferably, the Chinese medicine composition comprises, by weight: 40 parts of calamus, 40 parts of cycads, 60 parts of benzoin, and 100 parts of catechu.

[0013] Another aspect of the present invention provides a method for preparing a Chinese medicine composition, which specifically comprises the following steps:

[0014] S1. Weigh the Acorus calamus, dry it, crush it, add water, perform reflux extraction, and vacuum concentrate to remove the solvent to obtain the Acorus calamus extract.

[0015] S2. Weigh the iron club hammer, dry it, crush it, add water, reflux it for extraction, and vacuum concentrate it to remove the solvent to obtain the iron club hammer extract.

[0016] S3. Weigh benzoin, dry it, crush it, add 95% ethanol, let it stand for extraction, concentrate it under reduced pressure and dry it to obtain a benzoin extract.

[0017] S4, weighing catechu, drying, crushing, adding water, refluxing extraction, vacuum concentrating to remove the solvent, and obtaining catechu extract.

[0018] S5. Evenly mix the Acorus calamus extract, the Cyperus rotundus extract, the Styrax benzoin extract and the Catechu catechu extract to obtain the product.

[0019] Preferably, the temperature of the vacuum concentration in steps S1 to S4 is 55 to 60°C.

[0020] Preferably, the reflux extraction in step S1, step S2 and step S4 is performed 2 to 3 times, with a duration of 0.8 to 1.2 h per time.

[0021] Preferably, the number of static extractions in step S3 is 2 to 3 times, and the duration is 22 to 26 hours per time.

[0022] Preferably, the pulverizing in steps S1 to S4 is pulverizing into a coarsest powder of more than 10 mesh.

[0023] Another aspect of the present invention provides a use of the traditional Chinese medicine composition in preparing a medicine for treating hyperuricemia and / or gout.

[0024] Preferably, the medicine comprises one of a mixture, an ointment, a granule, a pill, a capsule, a syrup, a spray, a powder, a suspension, a tablet, a transdermal agent, and a gel.

[0025] Compared with the prior art, the present invention has the following beneficial effects:

[0026] (1) The present invention provides a traditional Chinese medicine composition comprising 20 to 60 parts of Acorus calamus, 30 to 50 parts of Cyperus rotundus, 20 to 60 parts of Benzoin, and 40 to 100 parts of Catechu, which significantly reduces the contents of XOD, SUA, CRE, BUN, TNF-α, IL-6, D-Lac, and DAO in the serum of a gout mouse model, and simultaneously reduces the renal index, and can effectively treat hyperuricemia and / or gout.

[0027] (2) The present invention provides a method for preparing the Chinese medicine composition, which can fully extract the active ingredients of Acorus calamus, Cyperus rotundus, Styrax benzoin, and Catechu species, thereby improving the utilization rate of the medicinal materials. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 This is the result diagram of the effect of different doses of Acorus calamus on the IL-1β content in the THP-1 cell gouty inflammation model;

[0029] Figure 2 This is the result of the effect of different doses of iron hammer on the IL-1β content in the THP-1 cell gouty inflammation model;

[0030] Figure 3 This is the result diagram of the effect of different doses of benzoin on the IL-1β content in the THP-1 cell gouty inflammation model;

[0031] Figure 4 This is the result diagram of the effect of different doses of catechins on the IL-1β content in the THP-1 cell gouty inflammation model;

[0032] Figure 5 This is a graph showing the effects of each group on the body weight of hyperuricemia rats. DETAILED DESCRIPTION

[0033] The present invention is further described below in conjunction with the examples. It should be noted that the following examples are only used to explain the present invention, but not to limit the present invention. The experimental methods used in the following examples are conventional methods unless otherwise specified. The materials, reagents, etc. used in the following examples, if not otherwise specified, can be obtained through commercial channels. Unless otherwise specified, the percentages mentioned in the present invention are all mass percentages.

[0034] The xanthine oxidase kit was purchased from Nanjing Jiancheng Bioengineering Institute (A002-1-1);

[0035] Sodium urate was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd. (M276559);

[0036] The uric acid (UA) test kit was purchased from Nanjing Jiancheng Bioengineering Institute (C012-2-1);

[0037] The creatinine (Cr) assay kit was purchased from Nanjing Jiancheng Bioengineering Institute (C011-2-1);

[0038] Urea nitrogen (BUN) test kit was purchased from Nanjing Jiancheng Bioengineering Institute (C013-2-1);

[0039] The xanthine oxidase (XOD) test kit was purchased from Nanjing Jiancheng Bioengineering Institute (A002-1-1);

[0040] D-Lactate ELISA kit was purchased from Jiangsu ELISA Industrial Co., Ltd. (MM-21239R1);

[0041] The diamine oxidase (DAO) ELISA research kit was purchased from Shanghai ELISA Biotechnology Co., Ltd. (CB10339-Ra);

[0042] Tumor necrosis factor (TNF-α) ELISA research kit was purchased from Jiangsu ELISA Industrial Co., Ltd. (MM-0180R1);

[0043] The interleukin (IL-6) ELISA research kit was purchased from Jiangsu ELISA Industrial Co., Ltd. (MM-0190R1).

[0044] Example 1

[0045] A Chinese medicine composition for treating hyperuricemia and gout is prepared, which specifically comprises the following steps:

[0046] S1. Weigh 40 g of Acorus calamus, dry at 60°C, mix and crush into a coarsest powder of 10 mesh, add 1 L of water, and perform three rounds of reflux extraction, each round lasting 1 hour. Then, the obtained extract is vacuum concentrated at 60°C until the solvent is removed to obtain an Acorus calamus extract.

[0047] S2. Weigh 40 g of the iron club hammer, dry it at 60°C, mix it, crush it into a coarsest powder of 10 mesh, add 1 L of water, and perform three rounds of reflux extraction, each round lasting 1 hour. Then, the obtained extract is vacuum concentrated at 60°C until the solvent is removed to obtain the iron club hammer extract.

[0048] S3. Weigh 60 g of benzoin, dry it at 60°C, mix it, and grind it into a coarsest powder with a mesh size of 10. Add 1 L of 95% ethanol, and extract it three times, each time for 24 hours. Concentrate the extract under reduced pressure and dry it to obtain a benzoin extract.

[0049] S4. Weigh 100 g of catechu, dry at 60° C., mix and crush into a coarsest powder of 10 mesh, add 1 L of water, and perform three rounds of reflux extraction, each round lasting 1 hour. Then, the obtained extract is vacuum concentrated at 60° C. until the solvent is removed to obtain a catechu extract.

[0050] S5. Mix the above four extracts evenly to obtain the product.

[0051] Example 2

[0052] A Chinese medicine composition for treating hyperuricemia and gout is prepared, which specifically comprises the following steps:

[0053] S1. Weigh 20 g of Acorus calamus, dry at 55°C, mix and crush into a coarsest powder of 10 mesh, add 1 L of water, and perform three rounds of reflux extraction, each round lasting 1.2 hours. Then, the obtained extract is concentrated under vacuum at 55°C until the solvent is removed, thereby obtaining an Acorus calamus extract.

[0054] S2. Weigh 30 g of the iron club hammer, dry it at 55°C, mix it, crush it into a coarsest powder of 10 mesh, add 1 L of water, and perform three rounds of reflux extraction, each round lasting 1.2 hours. Then, the obtained extract is vacuum concentrated at 55°C until the solvent is removed to obtain the iron club hammer extract.

[0055] S3. Weigh 20 g of benzoin, dry it at 55°C, mix it, and grind it into a coarsest powder with a mesh size of 10. Add 1 L of 95% ethanol, and extract it three times, each time for 26 hours. Concentrate the extract under reduced pressure and dry it to obtain a benzoin extract.

[0056] S4, weigh 80g of catechu, dry at 55°C, mix and crush into the coarsest powder of 10 mesh, add 1L of water, and perform three rounds of reflux extraction, each round lasting 1.2h. Then, the obtained extract is vacuum concentrated at 55°C until the solvent is removed to obtain catechu extract.

[0057] S5. Mix the above four extracts evenly to obtain the product.

[0058] Example 3

[0059] A Chinese medicine composition for treating hyperuricemia and gout is prepared, which specifically comprises the following steps:

[0060] S1. Weigh 60 g of Acorus calamus, dry at 65°C, mix and crush into a coarsest powder of 10 mesh, add 1 L of water, and perform three rounds of reflux extraction, each round lasting 0.8 h. Then, the obtained extract is vacuum concentrated at 65°C until the solvent is removed to obtain an Acorus calamus extract.

[0061] S2. Weigh 50 g of the iron club hammer, dry it at 65°C, mix it, crush it into a coarsest powder of 10 mesh, add 1 L of water, and perform three rounds of reflux extraction, each round lasting 0.8 h. Then, the obtained extract is vacuum concentrated at 65°C until the solvent is removed to obtain the iron club hammer extract.

[0062] S3. Weigh 40 g of benzoin, dry it at 65°C, mix it, and grind it into a coarsest powder with a mesh size of 10. Add 1 L of 95% ethanol, and extract it three times, each time for 22 hours. Concentrate the extract under reduced pressure and dry it to obtain a benzoin extract.

[0063] S4, weigh 60g of catechu, dry at 65°C, mix and crush into the coarsest powder of 10 mesh, add 1L of water, and perform three rounds of reflux extraction, each round lasting 0.8h. Then, the obtained extract is vacuum concentrated at 65°C until the solvent is removed to obtain catechu extract.

[0064] S5. Mix the above four extracts evenly to obtain the product.

[0065] The ingredients of Examples 1-9 are shown in Table 1. The preparation methods of Examples 4-9 are the same as that of Example 1.

[0066] Table 1 Chinese medicine composition formula

[0067] factor Acorus calamus(g) Iron hammer(g) Benzoin(g) Catechu (g) Example 1 40 40 60 100 Example 2 20 30 40 80 Example 3 20 50 40 100 Example 4 20 30 20 60 Example 5 40 40 60 60 Example 6 40 50 60 80 Example 7 60 30 20 80 Example 8 60 40 20 100 Example 9 60 50 40 60

[0068] Comparative Example 1

[0069] A Chinese medicine composition for treating hyperuricemia and gout is prepared, which specifically comprises the following steps:

[0070] S1. Weigh 40 g of Terminalia chebula, dry at 65°C, mix and crush into a coarsest powder of 10 mesh, add 1 L of water, and perform three rounds of reflux extraction, each round lasting 0.8 h. Then, the obtained extract is vacuum concentrated at 65°C until the solvent is removed to obtain Terminalia chebula extract.

[0071] S2. Weigh 40 g of Phyllanthus emblica, dry at 65° C., mix and crush into a coarsest powder of 10 mesh, add 1 L of water, and perform three rounds of reflux extraction, each round lasting 0.8 h. Then, the obtained extract is vacuum concentrated at 65° C. until the solvent is removed, to obtain the Phyllanthus emblica extract.

[0072] S3, weigh 40g of Radix Codonopsis pilosulae, dry at 65°C, mix and crush into the coarsest powder of 10 mesh, add 1L 70% ethanol, and perform three rounds of reflux extraction, each round lasting 0.8h. Then, the obtained extract is vacuum concentrated at 40°C until the solvent is removed to obtain Radix Codonopsis pilosulae extract.

[0073] S4, weigh 40g of Terminalia chebula, dry at 65°C, mix and crush into the coarsest powder of 10 mesh, add 1L of water, and perform three rounds of reflux extraction, each round lasting 0.8h. Then, the obtained extract is vacuum concentrated at 65°C until the solvent is removed to obtain Terminalia chebula extract.

[0074] S5. Weigh 40 g of costus root, dry at 65°C, mix and crush into a coarsest powder of 10 mesh, add 1 L of 70% ethanol, and perform three rounds of reflux extraction, each round lasting 0.8 h. Then, the obtained extract is vacuum concentrated at 40°C until the solvent is removed to obtain a costus root extract.

[0075] S6. Weigh 40 g of Cassia seed, dry at 65°C, mix and crush into a coarsest powder of 10 mesh, add 1 L of water, and perform three rounds of reflux extraction, each round lasting 0.8 h. Then, the obtained extract is vacuum concentrated at 65°C until the solvent is removed to obtain Cassia seed extract.

[0076] S7. Weigh 10 g of frankincense, dry it at 65°C, mix it, and grind it into a coarse powder with a mesh size of 10.

[0077] S8, weigh 40g of sunflower seeds, dry at 65°C, mix and crush into a coarsest powder of 10 mesh, add 1L of 70% ethanol, and perform three rounds of reflux extraction, each round lasting 0.8h. Then, the obtained extract is vacuum concentrated at 40°C until the solvent is removed to obtain a sunflower seed extract.

[0078] S9. Mix the above 8 extracts evenly to obtain the product.

[0079] Comparative Example 2

[0080] A Chinese medicine composition for treating hyperuricemia and gout is prepared, which specifically comprises the following steps:

[0081] S1. Weigh 40 g of cardamom, dry at 65°C, mix and crush into a coarsest powder of 10 mesh, add 1 L of water, and perform three rounds of reflux extraction, each round lasting 0.8 h. Then, the obtained extract is concentrated under vacuum at 65°C until the solvent is removed to obtain a cardamom extract.

[0082] S2. Weigh 40g of Amomum villosum, dry at 65°C, mix and crush into the coarsest powder of 10 mesh, add 1L of water, and perform three rounds of reflux extraction, each round lasting 0.8h. Then, the obtained extract is vacuum concentrated at 65°C until the solvent is removed to obtain Amomum villosum extract.

[0083] S3. Weigh 40 g of cloves, dry them at 65°C, mix and crush them into a coarsest powder of 10 mesh, add 1 L of water, and perform three rounds of reflux extraction, each round lasting 0.8 h. Then, the obtained extract is vacuum concentrated at 65°C until the solvent is removed to obtain a clove extract.

[0084] S4. Weigh 20 g of safflower, dry at 65°C, mix and crush into a coarsest powder of 10 mesh, add 1 L of 70% ethanol, and perform three rounds of reflux extraction, each round lasting 2 to 3 hours. Then, concentrate the obtained extract under vacuum at 40°C until the solvent is removed to obtain a safflower extract.

[0085] S5. Weigh 5g of travertine, mix and crush into coarse powder with a mesh size of 10.

[0086] S6. Mix the above five extracts evenly to obtain the product.

[0087] Comparative Example 3

[0088] A Chinese medicine composition for treating hyperuricemia and gout was prepared, which was basically the same as Example 1, except that 40 g of Acorus calamus was replaced with 40 g of Cassia seed.

[0089] Comparative Example 4

[0090] A Chinese medicine composition for treating hyperuricemia and gout was prepared, which was basically the same as Example 1, except that 40 g of the iron hammer was replaced with 40 g of Terminalia chebula.

[0091] Comparative Example 5

[0092] A Chinese medicine composition for treating hyperuricemia and gout was prepared, which was basically the same as Example 1, except that 60 g of benzoin was replaced with 60 g of Rhizoma Polygoni Multiflori.

[0093] Comparative Example 6

[0094] A Chinese medicine composition for treating hyperuricemia and gout was prepared, which was basically the same as Example 1, except that 100 g of catechu was replaced with 100 g of cardamom.

[0095] Comparative Example 7

[0096] A Chinese medicine composition for treating hyperuricemia and gout was prepared, which was basically the same as Example 1, with the only difference being that the amount of calamus in step S1 was changed to 70 g, the amount of iron hammer in step S2 was 70 g, the amount of benzoin in step S3 was changed to 70 g, and the amount of catechu in step S4 was changed to 30 g.

[0097] Comparative Example 8

[0098] A Chinese medicine composition for treating hyperuricemia and gout was prepared, which was basically the same as Example 1, with the only difference being that the amount of calamus in step S1 was changed to 20 g, the amount of iron hammer in step S2 was 10 g, the amount of benzoin in step S3 was changed to 60 g, and the amount of catechu in step S4 was changed to 150 g.

[0099] Comparative Example 9

[0100] A Chinese medicine composition for treating hyperuricemia and gout was prepared, which was basically the same as Example 1, with the only difference being that the amount of calamus in step S1 was changed to 10 g, the amount of iron hammer in step S2 was 50 g, the amount of benzoin in step S3 was changed to 80 g, and the amount of catechu in step S4 was changed to 50 g.

[0101] Experimental Example 1: In vitro detection of the ability of the Chinese medicine composition to inhibit xanthine oxidase

[0102] Experimental method: Use a xanthine oxidase kit to detect the in vitro xanthine oxidase inhibitory activity of Examples 1 to 9 and Comparative Examples 1 to 9. The experiment set up a blank group, a standard group and a sample group. In the blank group, 50 μL PBS was added to a 96-well plate. In the standard group, 25 μL PBS and 25 μL xanthine oxidase solution were added. In the experimental group, 25 μL of sample extract and 25 μL of xanthine oxidase were added. The absorbance (OD value) was measured at 530 nm. Each experiment was repeated 6 times. Xanthine oxidase inhibitory activity = (OD 标准组 -OD 空白组 ) / (OD 标准组 -OD 空白组 )-(OD 实验组 -OD 空白组 ) / (OD 标准组- -OD 空白组 ).

[0103] The results are shown in Table 3.

[0104] Table 3 Xanthine oxidase inhibition activity results

[0105]

[0106] As can be seen from the above table, the Chinese medicine compositions of Examples 1 to 9 of the present invention, including Acorus calamus, Psoralea corylifolia, Benzoin and Catechu, have much higher xanthine oxidase inhibitory activity than Comparative Examples 1 and 2, indicating that the Chinese medicine composition formula provided by the present invention has better therapeutic effects on the treatment of hyperuricemia and gout. It can also be seen from the comparison of the embodiments with Comparative Examples 3 to 6 that the formula of the present invention as a whole plays a synergistic role, and replacing any of the medicinal materials in the Chinese medicine composition of the present invention will weaken the efficacy to varying degrees. It can be seen from the difference between the embodiments and Comparative Examples 7 to 9 that the optimized ratio of the embodiments of the present invention can better inhibit the activity of xanthine oxidase, that is, it has a better therapeutic effect on hyperuricemia and gout.

[0107] Experimental Example 2: Detection of the effect of the Chinese medicine composition on the IL-1β content in the THP-1 cell gouty inflammation model

[0108] Experimental method: Human myeloid leukemia mononuclear cells (THP-1) were cultured in RPMI medium containing 10% fetal bovine serum in a cell culture incubator at 37°C and 5% CO2. THP-1 cells were inoculated into a culture dish (about 3.6×10 cells). 6 / dish), and when the cell density reached 70-80% under a microscope, 100ng / mL phorbol lipid (PMA) (37°C, 5% CO2 culture for 48h) was used to induce THP-1 cells to become adherent macrophages. After the cells adhered to the wall, the culture medium was discarded, and a new culture medium containing 100μg / mL sodium urate (hereinafter referred to as MSU) was added for incubation for 12h, and then the culture medium containing 100μg / mL of different drugs was replaced for further incubation for 24h, and the supernatant was collected. The IL-1β content in the supernatant was determined by Elisa. The lower the IL-1β content, the stronger the protective effect of the drug on the MSU-induced gout model, and the results are shown in Table 2.

[0109] Table 2 Effect of Chinese medicine combination on IL-1β content

[0110] factor IL-1β (pg / mL) factor IL-1β (pg / mL) Example 1 1841 Comparative Example 1 4672 Example 2 2156 Comparative Example 2 3958 Example 3 1985 Comparative Example 3 4580 Example 4 2722 Comparative Example 4 4878 Example 5 2412 Comparative Example 5 4109 Example 6 2378 Comparative Example 6 5132 Example 7 2258 Comparative Example 7 4975 Example 8 2075 Comparative Example 8 4078 Example 9 2878 Comparative Example 9 3443

[0111] As can be seen from the above table, the IL-1β content of Examples 1-9 is 1841-2878 pg / mL, which is much lower than 3443-5132 pg / mL of Comparative Examples 1-9. For Comparative Examples 1 and 2, their formulas are different from those of Examples 1-9 of the present invention, indicating that the Chinese medicine composition formula provided by the present invention has a better therapeutic effect on the treatment of hyperuricemia and / or gout. By comparing the results of Examples 1-9 with Comparative Examples 3-6, it can be seen that replacing any one of Acorus calamus, Psoralea corylifolia, Benzoin, and Catechu will reduce the efficacy of the present invention, proving that the synergistic effect of the four ingredients of the present invention will promote the therapeutic effect of the Chinese medicine composition provided by the present invention on hyperuricemia and / or gout. By comparing the results of Examples 1 to 9 with Comparative Examples 7 to 9, it can be seen that when the amount of catechu in the composition is too much or too little, its effect on reducing the IL-1β content will be weakened. At the same time, changes in the proportions of the four components of calamus, scutellaria baicalensis, benzoin and catechu will also have a certain impact on the effect of the present invention. The preferred proportion of 20 to 60 parts of calamus, 30 to 50 parts of scutellaria baicalensis, 20 to 60 parts of benzoin, and 40 to 100 parts of catechu in the present invention has better therapeutic effect on the treatment of hyperuricemia and / or gout.

[0112] Experimental Example 3: Optimization of the formula of Chinese medicine composition

[0113] 1. Detect the effect of different doses of Acorus calamus on the IL-1β content in THP-1 cell gouty inflammation model

[0114] Experimental method: A Chinese medicine composition of 30g of iron hammer, 60g of benzoin, 60g of catechu, and 20-100g of calamus was prepared to investigate the effects of different doses of calamus on the IL-1β content in the THP-1 cell gouty inflammation model. Figure 1 It can be seen that with the increase of Acorus calamus concentration, the IL-1β content in the THP-1 cell gouty inflammation model gradually increased. Therefore, 20g, 40g and 60g concentrations of Acorus calamus were selected for further experiments.

[0115] 2. Detect the effect of different doses of iron hammer on the IL-1β content of THP-1 cell gouty inflammation model

[0116] A Chinese medicinal composition of 10-50 g of Trichosanthes chinensis, 60 g of benzoin, 60 g of catechu, and 60 g of Acorus calamus was prepared to investigate the effects of different doses of Trichosanthes chinensis on the IL-1β content in a THP-1 cell gouty inflammation model. Figure 2 It can be seen that with the increase of the concentration of Acorus calamus, the IL-1β content in the THP-1 cell gouty inflammation model gradually decreased. Therefore, three high concentrations of Acorus calamus, 30g, 40g and 50g, were selected for further experiments.

[0117] 3. Detect the effect of different doses of benzoin on the IL-1β content in THP-1 cell gouty inflammation model

[0118] A Chinese medicine composition of 30g of iron hammer, 20-100g of benzoin, 60g of catechu, and 60g of calamus was prepared to investigate the effects of different doses of benzoin on the IL-1β content of the THP-1 cell gouty inflammation model. Figure 3 It can be seen that with the increase of benzoin concentration, the IL-1β content in the THP-1 cell gouty inflammation model gradually increased. Therefore, 20g, 40g and 60g benzoin concentrations were selected for further experiments.

[0119] 4. Detect the effect of different doses of catechins on the IL-1β content in THP-1 cell gouty inflammation model

[0120] A Chinese medicine composition of 30g of scutellaria baicalensis, 60g of benzoin, 20-100g of catechu, and 60g of calamus was prepared to investigate the effects of different doses of catechu on the IL-1β content of the THP-1 cell gouty inflammation model. Figure 4 It can be seen that with the increase of catechu concentration, the IL-1β content in the THP-1 cell gouty inflammation model gradually increased. Therefore, catechu concentrations of 60g, 80g and 100g were selected for further experiments.

[0121] The above experiments show that each component in the composition of the present invention has a certain therapeutic effect on the treatment of hyperuricemia and / or gout.

[0122] 5. Orthogonal experiment to optimize the formula

[0123] Table 3 Orthogonal optimization results of Chinese medicine composition formula

[0124]

[0125]

[0126] As shown in Table 3, when 40 grams of Acorus calamus, 40 grams of Trichosanthes ferox, 40 grams of Benzoin and 100 grams of Catechu, the IL-1β content in the THP-1 cell gouty inflammation model was the lowest. The influence of each factor on the IL-1β content from large to small was Catechu, Trichosanthes ferox, Acorus calamus and Benzoin. Therefore, from the optimization experiment, it can be seen that the best combination of the Chinese medicine composition for reducing IL-1β in the THP-1 cell gouty inflammation model is: 40 grams of Acorus calamus, 40 grams of Trichosanthes ferox, 60 grams of Benzoin and 100 grams of Catechu.

[0127] Experimental Example 4: Detection of the therapeutic effect of the Chinese medicine composition on hyperuricemia model rats induced by potassium oxonate

[0128] 1. Experimental animals: 48 SPF-grade male SD rats, 4 - 6 weeks old, with a body weight of (180.0 ± 10.0) g, were purchased from the Guangdong Provincial Medical Experimental Animal Center, and the animal license number is SCXK(Guangdong)2022 - 0002. All animals were housed in the Experimental Animal Center of Guangdong Pharmaceutical University, with the environmental temperature controlled at 20°C - 25°C, the light cycle of 12 hours per day, and the humidity of 40% - 70%. Adaptive feeding was carried out one week before the formal experiment began.

[0129] 2. Experimental methods: The above-mentioned 48 rats were randomly divided into 8 groups, namely the blank group, the model group, the positive group, the Acorus calamus group, the Aconitum pendulum group, the Benzoin group, the Catechu group, and the compound formula group, with 6 rats in each group. Except for the blank group, the other groups were given normal common feed and 10% fructose water from the 1st to the 6th week, and the fructose water was freely available for drinking. At 600 mg·kg -1 suspension of potassium oxonate was intragastrically administered every morning; in each drug-administered group from the 3rd to the 6th week, on the basis of intragastric administration of 600 mg·kg -1 suspension of potassium oxonate every morning, the corresponding drug was intragastrically administered, and the dosage of each mouse was 50 mg·kg -1 .

[0130] The positive drug control group (ALLO) was given 50 mg·kg -1 allopurinol solution. The Acorus calamus group was administered with Acorus calamus extract, the Aconitum pendulum group was administered with Aconitum pendulum extract, the Benzoin group was administered with Benzoin extract, the Catechu group was administered with Catechu extract, and the compound formula group was administered with the traditional Chinese medicine composition of Example 1.

[0131] During the experiment, the body weight of the rats was measured weekly, and the mental state, activity, hair color, diet, and urine and feces conditions of the animals were observed. Each group of rats was fasted for 8 h after the last administration, anesthetized with 10% chloral hydrate, and blood was taken through the abdominal aorta. After standing until stratification occurred, centrifugation was immediately carried out at 4000 r·min -1 for 15 min to separate the serum. The levels of serum uric acid (SUA), creatinine (CRE), blood urea nitrogen (BUN), xanthine oxidase (XOD), D-lactic acid (D-Lac), diamine oxidase (DAO), tumor necrosis factor (TNF-α), and interleukin (IL-6) were measured using a kit method.

[0132] All experimental data were expressed as Mean ± SD, and statistical analysis and graphing were performed using Graphpad Prism 8.0 and Spss. When the data of inter-group differences conformed to a normal distribution, one-way analysis of variance was used, and the test level α = 0.05. P < 0.05 indicated that the difference was statistically significant, and P < 0.01 indicated that the difference was extremely statistically significant.

[0133] 3. Experimental results:

[0134] During the modeling period, the rats in each group had normal fur color, good spirits, and normal diet, urination and defecation. The weight of the rats in each group showed an overall upward trend. Figure 5 As shown, there was no statistical difference among the groups (P>0.05).

[0135] The serum XOD and SUA levels of the rats in the model group were significantly higher than those in the blank group (P<0.01), indicating that the model was successfully constructed. The serum XOD and SUA levels of the rats in the compound formula group were significantly lower than those in the Acorus calamus group, the iron hammer group, the benzoin group, and the catechu group (P<0.01 or P<0.05), as shown in Table 4.

[0136] Table 4 Comparison of XOD and SUA in each group (n=6)

[0137]

[0138] Note: Compared with the model group, * P<0.05, ** P<0.01; compared with the compound formula group, # P<0.05, ## P<0.01.

[0139] The serum CRE, BUN and renal index of rats in the model group were significantly higher than those in the control group (P<0.01), indicating that the model was successfully constructed. The formula for calculating the renal index was: renal index = rat kidney weight / rat body weight × 100%. The serum CRE and BUN levels of rats in the compound formula group were significantly lower than those in the Acorus calamus group, the iron hammer group, the benzoin group, and the catechu group (P<0.01 or P<0.05), and the renal index of the compound formula group was lower than that of the Acorus calamus group (P<0.05). The results are shown in Table 5.

[0140] Table 5 Comparison of CRE, BUN and renal index in each group (n=6)

[0141]

[0142]

[0143] Note: Compared with the model group, * P<0.05, ** P<0.01; compared with the compound formula group, # P<0.05, ## P<0.01.

[0144] The levels of serum TNF-α, IL-6, D-Lac and DAO in the model group were significantly higher than those in the control group (P<0.01), indicating that the model was successfully constructed. The levels of serum TNF-α, IL-6, D-Lac and DAO in the compound formula group were significantly lower than those in the calamus group, the iron hammer group, the benzoin group and the catechu group (P<0.01 or P<0.05), as shown in Table 6.

[0145] Table 6 Comparison of TNF-α, IL-6, D-Lac and DAO in each group (n=6)

[0146]

[0147] Note: Compared with the model group, * P<0.05, ** P<0.01; compared with the compound formula group, # P<0.05, ## P<0.01.

[0148] In summary, from the above results, it can be seen that the composite formula group administered using Example 1 of the present invention has a significantly better effect on reducing serum XOD and SUA, serum CRE and BUN, serum TNF-α, IL-6, D-Lac and DAO than single administration, that is, when the dosage is the same, the composite formula is better than single administration. The calamus, scutellaria, benzoin and catechu provided by the present invention can synergistically enhance the effect and have a good effect on the treatment of hyperuricemia and / or gout.

Claims

1. A Chinese medicine composition, characterized in that: The invention comprises the following ingredients by weight: 20-60 parts of calamus, 30-50 parts of iron hammer, 20-60 parts of benzoin and 40-100 parts of catechu.

2. The Chinese medicine composition according to claim 1, characterized in that: The invention comprises the following ingredients by weight: 40 parts of calamus, 40 parts of iron hammer, 60 parts of benzoin and 100 parts of catechu.

3. The method for preparing the Chinese medicine composition according to any one of claims 1 to 2, characterized in that: The following steps are involved: S1, weighing Acorus calamus, drying, crushing, adding water, refluxing extraction, and vacuum concentrating to remove the solvent to obtain Acorus calamus extract; S2, weighing the iron club hammer, drying, crushing, adding water, reflux extraction, vacuum concentration to remove the solvent, to obtain the iron club hammer extract; S3, weighing benzoin, drying, crushing, adding 95% ethanol, standing for extraction, concentrating and drying under reduced pressure to obtain a benzoin extract; S4, weighing catechu, drying, crushing, adding water, refluxing extraction, vacuum concentrating to remove the solvent, and obtaining catechu extract; S5. Evenly mix the Acorus calamus extract, the Cyperus rotundus extract, the Styrax benzoin extract and the Catechu catechu extract to obtain the product.

4. The preparation method according to claim 3, characterized in that: The temperature of the vacuum concentration in steps S1 to S4 is 55 to 60°C.

5. The preparation method according to claim 3, characterized in that: The number of reflux extractions in step S1, step S2 and step S4 is 2 to 3 times, and the duration is 0.8 to 1.2 hours per time.

6. The preparation method according to claim 3, characterized in that: The number of static extractions in step S3 is 2 to 3 times, and the duration is 22 to 26 hours per time.

7. The preparation method according to claim 3, characterized in that: The pulverization in steps S1 to S4 is pulverizing into the coarsest powder of 10 mesh.

8. Use of the Chinese medicine composition according to any one of claims 1 to 2 in the preparation of a medicament for treating hyperuricemia and / or gout.

9. The use according to claim 8, characterized in that The medicine includes one of a mixture, an ointment, a granule, a pill, a capsule, a syrup, a spray, a powder, a suspension, a tablet, a transdermal agent, and a gel.

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