A traditional Chinese medicine composition, its preparation method and uses

Abstract

This invention belongs to the field of traditional Chinese medicine technology, specifically relating to a traditional Chinese medicine composition, its preparation method, and its uses. By mass parts, the raw materials of the traditional Chinese medicine composition include: 20-60 parts of Acorus calamus, 30-50 parts of Citrus aurantium, 20-60 parts of Benzoin, and 40-100 parts of Catechu. The traditional Chinese medicine composition of this invention has simple and natural components, and clear efficacy. It can effectively reduce the levels of XOD, SUA, CRE, BUN, TNF-α, IL-6, D-Lac, and DAO in the serum of a mouse model of gout, and significantly reduce the renal index, demonstrating good efficacy in treating hyperuricemia and / or gout.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine technology, specifically relating to a traditional Chinese medicine composition, its preparation method, and its uses. Background Technology

[0002] Hyperuricemia is a common metabolic disorder associated with a variety of serious diseases. It is caused by excessive uric acid production or insufficient excretion by the kidneys and intestines. It is not only a risk factor for gout but also closely related to the onset and progression of hypertension, diabetes, and kidney disease.

[0003] The general treatment strategy for hyperuricemia focuses on using xanthine oxidase inhibitors, such as allopurinol and febuxostat, to lower uric acid levels, and 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 allergic reactions, and nephrotoxicity, often limit their clinical use. Therefore, finding more effective and non-toxic natural products to treat hyperuricemia is crucial.

[0004] Over the years, Tibetan medicine has accumulated a large number of drugs for treating hyperuricemia. However, traditional Tibetan medicine has complex ingredients, unclear mechanisms of action, and difficulty in identifying effective components. Therefore, it is necessary to develop a traditional Chinese medicine composition with simple ingredients, clear efficacy, and good therapeutic effect for treating 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 with simple ingredients, clear efficacy, and good therapeutic effect for the treatment of hyperuricemia and / or gout. It significantly reduces the levels of XOD, SUA, CRE, BUN, TNF-α, IL-6, D-Lac, and DAO in the serum of a mouse model of gout, while also reducing the renal index, and can effectively treat hyperuricemia and / or gout.

[0006] To achieve the above objectives, the present invention provides a traditional Chinese medicine composition comprising, by weight, the following ingredients: 20-60 parts of Acorus calamus, 30-50 parts of Cibotium barometz, 20-60 parts of Benzoin, and 40-100 parts of Catechu.

[0007] The water calamus used in this invention is the rhizome of *Acorus calamus* L., a plant in the Acorusceae family. It has the effects of clearing the orifices, resolving phlegm, and regulating qi, and is used as an assistant herb. It is commonly used for phlegm-induced coma, stroke, epilepsy, palpitations and forgetfulness, tinnitus and deafness, food stagnation and abdominal pain, dysentery and diarrhea, rheumatic pain, eczema, and scabies.

[0008] The iron rod hammer used in this invention is the dried tuberous root of Aconitum pendulum Busch., a plant of the Ranunculaceae family. It has the effects of dispelling wind and dampness, dispersing blood stasis and relieving pain. It is used as an adjuvant medicine to treat injuries from falls and blows, fractures, rheumatic back pain, frostbite and other symptoms.

[0009] The benzoin used in this invention is the dried resin of *Styrax tonkinensis* (Pierre) Craib ex Hart., a plant in the Styraceae family. Main uses: clears the mind and spirit, promotes qi and blood circulation, relieves pain; used as an adjuvant. Indications: used for stroke with phlegm syncope, sudden syncope due to qi stagnation, coma due to sudden illness, abdominal pain, postpartum hemorrhage, and infantile convulsions.

[0010] The catechu used in this invention is the dried decoction of the peeled branches and twigs of the legume Acacia catechu (Lf) Willci. It has the effects of promoting blood circulation and relieving pain, stopping bleeding and promoting tissue regeneration, astringing and healing sores, clearing the lungs and resolving phlegm, and is the principal ingredient. It is used for injuries from falls and blows, external bleeding, hematemesis, epistaxis, non-healing sores, eczema, damp sores, and cough due to lung heat.

[0011] The traditional Chinese medicine composition provided by this invention, which includes calamus, iron rod, benzoin, and catechu, is rationally combined. The four traditional Chinese medicine components work synergistically to achieve good therapeutic effects on hyperuricemia and / or gout.

[0012] Preferably, the traditional Chinese medicine composition comprises, by weight, 40 parts of Acorus calamus, 40 parts of Cibotium barometz, 60 parts of Benzoin, and 100 parts of Catechu.

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

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

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

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

[0017] S4. Weigh the catechu, dry it, pulverize it, add water, reflux to extract it, and vacuum concentrate it to remove the solvent to obtain the catechu extract.

[0018] S5. Mix the calamus extract, iron rod extract, benzoin extract, and catechu extract evenly to obtain the final product.

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

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

[0021] Preferably, the static extraction in step S3 is performed 2 to 3 times, with a duration of 22 to 26 hours per extraction.

[0022] Preferably, the pulverization in steps S1 to S4 is pulverization into the coarsest powder passing through 10 mesh.

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

[0024] Preferably, the drug includes one of the following: compound preparation, ointment, granule, pill, capsule, syrup, spray, powder, suspension, tablet, transdermal preparation, and gel.

[0025] Compared with the prior art, the beneficial effects of the present invention are as follows:

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

[0027] (2) The present invention provides a method for preparing the traditional Chinese medicine composition, which can fully extract the active ingredients of Acorus calamus, calamus, benzoin and catechu, and improve the utilization rate of the medicinal materials. Attached Figure Description

[0028] Figure 1 Figure showing the effect of different doses of Acorus calamus on IL-1β levels in a THP-1 cell gouty inflammation model;

[0029] Figure 2 Figure showing the effect of different doses of iron bar on IL-1β content in a THP-1 cell gouty inflammation model;

[0030] Figure 3 Figure showing the effect of different doses of benzoin on IL-1β levels in a THP-1 cell gouty inflammation model;

[0031] Figure 4 Figure showing the effect of different doses of catechu on IL-1β levels in a THP-1 cell gouty inflammation model;

[0032] Figure 5The graph shows the effect of each group on the body weight of rats with high uric acid. Detailed Implementation

[0033] The present invention will be further described below with reference to embodiments. It should be noted that the following embodiments are only for explaining the present invention and are not intended to limit the present invention. Unless otherwise specified, the experimental methods used in the following embodiments are all conventional methods. Unless otherwise specified, the materials and reagents used in the following embodiments are commercially available. Unless otherwise specified, all percentages mentioned in the present invention are mass percentages.

[0034] The xanthine oxidase kit was purchased from Nanjing Jiancheng Biotechnology 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 Biotechnology Institute (C011-2-1).

[0038] The blood 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 Enzyme Immunization Industry Co., Ltd. (MM-21239R1).

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

[0042] The tumor necrosis factor (TNF-α) ELISA research kit was purchased from Jiangsu Enzyme Immunoassay Co., Ltd. (MM-0180R1).

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

[0044] Example 1

[0045] The preparation of a traditional Chinese medicine composition for treating hyperuricemia and gout specifically includes the following steps:

[0046] S1. Weigh 40g of sweet flag (Acorus calamus), dry it at 60℃, mix and pulverize it into the coarsest powder passing through 10 mesh, add 1L of water, and perform three rounds of reflux extraction, each round lasting 1 hour. Then, concentrate the obtained extract under vacuum at 60℃ until the solvent is removed to obtain sweet flag extract.

[0047] S2. Weigh 40g of iron rod, dry it at 60℃, mix and grind it into the coarsest powder passing through 10 mesh, add 1L of water, and perform three rounds of reflux extraction, each round lasting 1 hour. Then, concentrate the obtained extract under vacuum at 60℃ until the solvent is removed to obtain the iron rod extract.

[0048] S3. Weigh 60g of benzoin, dry it at 60℃, mix and pulverize it into the coarsest powder passing through 10 mesh, add 1L of 95% ethanol, let it stand for extraction 3 times, 24 hours each time, concentrate and dry the extract under reduced pressure to obtain benzoin extract.

[0049] S4. Weigh 100g of catechu, dry it at 60℃, mix and grind it into the coarsest powder passing through a 10-mesh sieve, add 1L of water, and perform three rounds of reflux extraction, each round lasting 1 hour. Then, concentrate the obtained extract under vacuum at 60℃ until the solvent is removed to obtain the catechu extract.

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

[0051] Example 2

[0052] The preparation of a traditional Chinese medicine composition for treating hyperuricemia and gout specifically includes the following steps:

[0053] S1. Weigh 20g of sweet flag (Acorus calamus), dry it at 55℃, mix and pulverize it into the coarsest powder passing through a 10-mesh sieve, add 1L of water, and perform three rounds of reflux extraction, each round lasting 1.2 hours. Then, concentrate the obtained extract under vacuum at 55℃ until the solvent is removed to obtain sweet flag extract.

[0054] S2. Weigh 30g of iron bark, dry it at 55℃, mix and grind it into the coarsest powder passing through 10 mesh, add 1L of water, and perform three rounds of reflux extraction, each round lasting 1.2h. Then, concentrate the obtained extract under vacuum at 55℃ until the solvent is removed to obtain the iron bark extract.

[0055] S3. Weigh 20g of benzoin, dry it at 55℃, mix and grind it into the coarsest powder passing through 10 mesh, add 1L of 95% ethanol, let it stand for extraction 3 times, 26 hours each time, concentrate and dry the extract under reduced pressure to obtain benzoin extract.

[0056] S4. Weigh 80g of catechu, dry it at 55℃, mix and grind it into the coarsest powder passing through a 10-mesh sieve, add 1L of water, and perform three rounds of reflux extraction, each round lasting 1.2 hours. Then, concentrate the obtained extract under vacuum at 55℃ until the solvent is removed to obtain the catechu extract.

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

[0058] Example 3

[0059] The preparation of a traditional Chinese medicine composition for treating hyperuricemia and gout specifically includes the following steps:

[0060] S1. Weigh 60g of sweet flag (Acorus calamus), dry it at 65℃, mix and pulverize it into the coarsest powder passing through a 10-mesh sieve, add 1L of water, and perform three rounds of reflux extraction, each round lasting 0.8h. Then, concentrate the obtained extract under vacuum at 65℃ until the solvent is removed to obtain sweet flag extract.

[0061] S2. Weigh 50g of iron bark, dry it at 65℃, mix and grind it into the coarsest powder passing through 10 mesh, add 1L of water, and perform three rounds of reflux extraction, each round lasting 0.8h. Then, concentrate the obtained extract under vacuum at 65℃ until the solvent is removed to obtain the iron bark extract.

[0062] S3. Weigh 40g of benzoin, dry it at 65℃, mix and pulverize it into the coarsest powder passing through 10 mesh, add 1L of 95% ethanol, let it stand for extraction 3 times, 22 hours each time, concentrate and dry the extract under reduced pressure to obtain benzoin extract.

[0063] S4. Weigh 60g of catechu, dry it at 65℃, mix and grind it into the coarsest powder passing through 10 mesh, add 1L of water, and perform three rounds of reflux extraction, each round lasting 0.8h. Then, concentrate the obtained extract under vacuum at 65℃ until the solvent is removed to obtain catechu extract.

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

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

[0066] Table 1. Formulation of Traditional Chinese Medicine Compositions.

[0067]

[0068] Comparative Example 1

[0069] The preparation of a traditional Chinese medicine composition for treating hyperuricemia and gout specifically includes the following steps:

[0070] S1. Weigh 40g of Terminalia chebula, dry it at 65℃, mix and pulverize it into the coarsest powder passing through 10 mesh, add 1L of water, and perform three rounds of reflux extraction, each round lasting 0.8h. Then, concentrate the obtained extract under vacuum at 65℃ until the solvent is removed to obtain Terminalia chebula extract.

[0071] S2. Weigh 40g of Phyllanthus emblica, dry it at 65℃, mix and pulverize it into the coarsest powder passing through a 10-mesh sieve, add 1L of water, and perform three rounds of reflux extraction, each round lasting 0.8h. Then, concentrate the obtained extract under vacuum at 65℃ until the solvent is removed to obtain Phyllanthus emblica extract.

[0072] S3. Weigh 40g of *Tinospora sinensis*, dry it at 65℃, mix and pulverize it into the coarsest powder passing through a 10-mesh sieve. Add 1L of 70% ethanol and perform three rounds of reflux extraction, each round lasting 0.8h. Then, concentrate the obtained extract under vacuum at 40℃ until the solvent is removed to obtain *Tinospora sinensis* extract.

[0073] S4. Weigh 40g of Terminalia chebula, dry it at 65℃, mix and pulverize it into the coarsest powder passing through a 10-mesh sieve, add 1L of water, and perform three rounds of reflux extraction, each round lasting 0.8h. Then, concentrate the obtained extract under vacuum at 65℃ until the solvent is removed to obtain Terminalia chebula extract.

[0074] S5. Weigh 40g of Costus root, dry it at 65℃, mix and pulverize it into the coarsest powder passing through a 10-mesh sieve. Add 1L of 70% ethanol and perform three rounds of reflux extraction, each round lasting 0.8h. Then, concentrate the obtained extract under vacuum at 40℃ until the solvent is removed to obtain Costus root extract.

[0075] S6. Weigh 40g of cassia seeds, dry them at 65℃, mix and pulverize them into the coarsest powder passing through a 10-mesh sieve, add 1L of water, and perform three rounds of reflux extraction, each round lasting 0.8h. Then, concentrate the obtained extract under vacuum at 65℃ until the solvent is removed to obtain cassia seed extract.

[0076] S7. Weigh 10 g of frankincense, dry it at 65℃, mix it, and grind it into coarse powder that passes through 10 mesh.

[0077] S8. Weigh 40g of okra seeds, dry them at 65℃, mix and grind them into the coarsest powder passing through a 10-mesh sieve. Add 1L of 70% ethanol and perform three rounds of reflux extraction, each round lasting 0.8h. Then, concentrate the obtained extract under vacuum at 40℃ until the solvent is removed to obtain okra seed extract.

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

[0079] Comparative Example 2

[0080] The preparation of a traditional Chinese medicine composition for treating hyperuricemia and gout specifically includes the following steps:

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

[0082] S2. Weigh 40g of cardamom, dry it at 65℃, mix and grind it into the coarsest powder passing through 10 mesh, add 1L of water, and perform three rounds of reflux extraction, each round lasting 0.8h. Then, concentrate the obtained extract under vacuum at 65℃ until the solvent is removed to obtain cardamom extract.

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

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

[0085] S5. Weigh 5g of travertine, mix and grind it into coarse powder that passes through 10 mesh.

[0086] S6. Mix the above 5 extracts evenly to obtain the final product.

[0087] Comparative Example 3

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

[0089] Comparative Example 4

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

[0091] Comparative Example 5

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

[0093] Comparative Example 6

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

[0095] Comparative Example 7

[0096] A traditional Chinese medicine composition for treating hyperuricemia and gout was prepared, which was basically the same as that in Example 1, except that: the amount of Acorus tatarinowii in step S1 was changed to 70g, the amount of Cibotium barometz in step S2 was 70g, the amount of Benzoin in step S3 was changed to 70g, and the amount of Catechu in step S4 was changed to 30g.

[0097] Comparative Example 8

[0098] A traditional Chinese medicine composition for treating hyperuricemia and gout was prepared, which was basically the same as that in Example 1, except that: the amount of Acorus tatarinowii in step S1 was changed to 20g, the amount of Cibotium barometz in step S2 was 10g, the amount of Benzoin in step S3 was changed to 60g, and the amount of Catechu in step S4 was changed to 150g.

[0099] Comparative Example 9

[0100] A traditional Chinese medicine composition for treating hyperuricemia and gout was prepared, which was basically the same as that in Example 1, except that: the amount of Acorus tatarinowii in step S1 was changed to 10g, the amount of Cibotium barometz in step S2 was 50g, the amount of Benzoin in step S3 was changed to 80g, and the amount of Catechu in step S4 was changed to 50g.

[0101] Experimental Example 1: In vitro detection of the inhibitory activity of traditional Chinese medicine composition against xanthine oxidase

[0102] Experimental Methods: The in vitro xanthine oxidase inhibitory activity of Examples 1-9 and Comparative Examples 1-9 was detected using a xanthine oxidase kit. The experiment included a blank group, a standard group, and a sample group. For the blank group, 50 μL of PBS was added to each well of a 96-well plate. For the standard group, 25 μL of PBS and 25 μL of xanthine oxidase solution were added. For 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 standard group - OD blank group) / (OD standard group - OD blank group) - (OD experimental group - OD blank group) / (OD standard group - OD blank group).

[0103] The results are shown in Table 2-1.

[0104] Table 2-1 Results of xanthine oxidase inhibitory activity

[0105]

[0106] As shown in the table above, the herbal compositions of Acorus calamus, Haliotis diversicolor, Benzoin, and Catechu in Examples 1-9 of this invention exhibit significantly higher inhibitory activity against xanthine oxidase than those in Comparative Examples 1 and 2, indicating that the herbal composition formulations provided by this invention have better therapeutic effects on hyperuricemia and gout. A comparison between the examples and Comparative Examples 3-6 also shows that the formulations of this invention, as a whole, exert a synergistic effect. Replacing any one of the herbs in the herbal composition of this invention will weaken the efficacy to varying degrees. The differences between the examples and Comparative Examples 7-9 demonstrate that the optimized formulations of the examples of this invention can better inhibit xanthine oxidase activity, thus achieving a better therapeutic effect on hyperuricemia and gout.

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

[0108] Experimental Methods: Human myeloid leukemia mononuclear cells (THP-1) were cultured in RPMI medium containing 10% fetal bovine serum at 37°C in a 5% CO2 cell culture incubator. THP-1 cells were seeded into culture dishes (approximately 3.6 × 10⁶ cells / dish). When the cell density reached 70-80% under a microscope, THP-1 cells were induced to become adherent macrophages by incubating with 100 ng / mL phorbol ester (PMA) at 37°C in 5% CO2 for 48 h. After cell adhesion, the culture medium was discarded, and fresh medium containing 100 μg / mL sodium urate (MSU) was added and incubated for 12 h. This was then replaced with medium containing 100 μg / mL of different drugs and incubated for another 24 h. The supernatant was collected, and the IL-1β content in the supernatant was determined using ELISA. The lower the IL-1β content, the stronger the protective effect of the drug on the MSU-induced gout model, as shown in Table 2-2.

[0109] Table 2-2 Results of the Influence of Traditional Chinese Medicine Compositions on IL-1β Content

[0110]

[0111] As shown in the table above, the IL-1β content of Examples 1-9 ranges from 1841 to 2878 pg / mL, which is significantly lower than the 3443 to 5132 pg / mL of Comparative Examples 1-9. Comparative Examples 1 and 2 have different formulations than Examples 1-9 of this invention, indicating that the traditional Chinese medicine composition provided by this invention has a better therapeutic effect on hyperuricemia and / or gout. A comparison of the results of Examples 1-9 with Comparative Examples 3-6 shows that replacing any one of the following—Acorus calamus, cycad, benzoin, and catechu—decreases the efficacy of this invention, demonstrating that the synergistic effect of the four components of this invention promotes the therapeutic effect of the traditional Chinese medicine composition provided by this invention on hyperuricemia and / or gout. A comparison of the results of Examples 1-9 and Comparative Examples 7-9 shows that both excessive and insufficient catechu in the composition weaken its effect on reducing IL-1β levels. Furthermore, changes in the proportions of the four components—Acorus calamus, Hedyotis diffusa, Benzoin, and catechu—also affect the efficacy of the invention. The preferred proportions of 20-60 parts Acorus calamus, 30-50 parts Hedyotis diffusa, 20-60 parts Benzoin, and 40-100 parts catechu have a better therapeutic effect on hyperuricemia and / or gout.

[0112] Experimental Example 3: Optimization of Traditional Chinese Medicine Composition Formulation

[0113] 1. To investigate the effect of different doses of Acorus calamus on IL-1β levels in a THP-1 cell gouty inflammation model.

[0114] Experimental Methods: A traditional Chinese medicine composition consisting of 30 g of *Imperata cylindrica*, 60 g of benzoin, 60 g of catechu, and 20-100 g of *Acorus calamus* was prepared. The effect of different doses of *Acorus calamus* on the IL-1β content in a THP-1 cell gouty inflammation model was investigated. Figure 1 It was found that as the concentration of calamus increased, the IL-1β content in the THP-1 cell gouty inflammation model gradually increased. Therefore, 20g, 40g and 60g concentrations of calamus were selected for further experiments.

[0115] 2. Detection of the effect of different doses of iron bar on IL-1β content in a THP-1 cell gouty inflammation model.

[0116] A traditional Chinese medicine composition consisting of 10-50 g of *Imperata cylindrica*, 60 g of benzoin, 60 g of catechu, and 60 g of *Acorus calamus* was prepared. The effects of different doses of *Imperata cylindrica* on the IL-1β content in a THP-1 cell gouty inflammation model were investigated. Figure 2 It was found that as the concentration of *Acorus calamus* increased, 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. Effects of different doses of benzoin on IL-1β levels in a THP-1 cell gouty inflammation model.

[0118] A traditional Chinese medicine composition consisting of 30g of *Impatiens balsamina*, 20-100g of benzoin, 60g of catechu, and 60g of *Acorus calamus* was prepared. The effect of different doses of benzoin on the IL-1β content in a THP-1 cell gouty inflammation model was investigated. Figure 3 It was found that as the concentration of benzoin increased, the IL-1β content in the THP-1 cell gouty inflammation model gradually increased. Therefore, benzoin concentrations of 20g, 40g, and 60g were selected for further experiments.

[0119] 4. To investigate the effect of different doses of catechu on IL-1β levels in a THP-1 cell gouty inflammation model.

[0120] A traditional Chinese medicine composition consisting of 30g of *Impatiens balsamina*, 60g of benzoin, 20-100g of catechu, and 60g of *Acorus calamus* was prepared. The effect of different doses of catechu on the IL-1β content in a THP-1 cell gouty inflammation model was investigated. Figure 4 It was found that as the concentration of catechu increased, the IL-1β content in the THP-1 cell gouty inflammation model gradually increased. Therefore, 60g, 80g and 100g concentrations of catechu 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 hyperuricemia and / or gout.

[0122] 5. Orthogonal experiment to optimize the formula

[0123] Table 3. Results of orthogonal optimization of traditional Chinese medicine composition formulations

[0124]

[0125] Table 3 shows that the lowest IL-1β content was observed in the THP-1 cell gouty inflammation model when the concentrations of Acorus tatarinowii (40g), Cibotium barometz (40g), Benzoin (40g), and Catechu (100g) were used. The order of influence of each factor on IL-1β content from largest to smallest was: Catechu, Cibotium barometz, Acorus tatarinowii, and Benzoin. Therefore, the optimal combination of traditional Chinese medicine ingredients to reduce IL-1β in the THP-1 cell gouty inflammation model is determined by the following composition: Acorus tatarinowii (40g), Cibotium barometz (40g), Benzoin (60g), and Catechu (100g).

[0126] Experiment Example 4: Detection of the therapeutic effect of a traditional Chinese medicine composition on a rat model of hyperuricemia induced by potassium oxonate.

[0127] 1. Experimental animals: 48 SPF-grade male SD rats, 4 - 6 weeks old, weighing (180.0 ± 10.0) g, were purchased from the Guangdong Provincial Medical Experimental Animal Center, with the animal license number 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 official start of the experiment.

[0128] 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 Styrax group, the Catechu group, and the compound formula group, with 6 rats in each group. Except for the blank group, the remaining groups were given normal common feed and 10% fructose water from week 1 to week 6, and the fructose water was freely available for drinking. At 600 mg·kg-1 of potassium oxonate suspension was intragastrically administered every morning; in each drug-administered group from week 3 to week 6, on the basis of intragastrically administering 600 mg·kg-1 of potassium oxonate suspension every morning, the corresponding drugs were intragastrically administered, and the dosage of each mouse was 50 mg·kg-1.

[0129] The positive drug control group (ALLO) was given 50 mg·kg-1 of allopurinol solution. The Acorus calamus group was administered with Acorus calamus extract, the Aconitum pendulum group was administered with Aconitum pendulum extract, the Styrax group was administered with Styrax 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.

[0130] During the experiment, the body weight of the rats was measured weekly, and the mental state, activity, hair color, diet, and urine and feces 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.

[0131] 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 ANOVA was used, with the test level α = 0.05. P < 0.05 was considered statistically significant, and P < 0.01 was considered extremely significantly statistically significant.

[0132] 3. Experimental results:

[0133] During the modeling period, the hair color of the rats in each group was normal, the mental state was good, and the diet, urine, and defecation were all normal. The overall body weight of the rats in each group showed an increasing trend, such as Figure 5 As shown, there were no statistically significant differences between the groups (P > 0.05).

[0134] The serum XOD and SUA levels in the model group rats were significantly higher than those in the blank group (P<0.01), indicating successful model establishment. The serum XOD and SUA levels in the compound formula group rats were significantly lower than those in the calamus group, iron rod group, benzoin group, and catechu group (P<0.01 or P<0.05), as shown in Table 4.

[0135] Table 4 Comparison results of XOD and SUA for each group (n=6)

[0136]

[0137] Note: Compared with the model group, *P<0.05, **P<0.01; compared with the compound formulation group, #P<0.05, ##P<0.01.

[0138] Serum CRE and BUN levels, as well as the kidney index, were significantly higher in the model group than in the control group (P<0.01), indicating successful model establishment. The kidney index was calculated as: Kidney Index = (Rats' Kidney Weight / Rats' Body Weight) × 100%. Serum CRE and BUN levels in the compound formula group were significantly lower than those in the calamus, iron rod, benzoin, and catechu groups (P<0.01 or P<0.05), and the kidney index in the compound formula group was lower than that in the calamus group (P<0.05). The results are shown in Table 5.

[0139] Table 5. Comparison of CRE, BUN, and Kidney Index among Groups (n=6)

[0140]

[0141] Note: Compared with the model group, *P<0.05, **P<0.01; compared with the compound formulation group, #P<0.05, ##P<0.01.

[0142] The serum levels of TNF-α, IL-6, D-Lac, and DAO in the model group rats were significantly higher than those in the control group (P<0.01), indicating successful model establishment. The serum levels of TNF-α, IL-6, D-Lac, and DAO in the compound formula group rats were significantly lower than those in the calamus group, ironwood group, benzoin group, and catechu group (P<0.01 or P<0.05), as shown in Table 6.

[0143] Table 6. Comparison of TNF-α, IL-6, D-Lac, and DAO levels in each group (n=6)

[0144]

[0145] Note: Compared with the model group, *P<0.05, **P<0.01; compared with the compound formulation group, #P<0.05, ##P<0.01.

[0146] In summary, the results show that the compound formulation group administered according to Example 1 of this 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 the single administration. That is, under the same dosage, the compound formulation is more effective than the single administration. The calamus, iron rod, benzoin and catechu provided by this invention can synergistically enhance the effect and have a good effect on the treatment of hyperuricemia and / or gout.

Claims

1. A traditional Chinese medicine composition, characterized in that, It is made from 20-60 parts by weight of sweet flag, 30-50 parts of iron rod, 20-60 parts of benzoin, and 40-100 parts of catechu. The preparation method of the traditional Chinese medicine composition is as follows: S1. Weigh the calamus, dry it, pulverize it, add water, reflux to extract it, and vacuum concentrate it to remove the solvent to obtain the calamus extract. S2. Weigh the iron rod, dry it, crush it, add water, reflux to extract it, and vacuum concentrate it to remove the solvent to obtain the iron rod extract. S3. Weigh out benzoin, dry it, pulverize it, add 95% ethanol, let it stand to extract, concentrate and dry it under reduced pressure to obtain benzoin extract; S4. Weigh the catechu, dry it, pulverize it, add water, reflux to extract it, and vacuum concentrate it to remove the solvent to obtain the catechu extract. S5. Mix the calamus extract, iron rod extract, benzoin extract, and catechu extract evenly to obtain the final product.

2. The traditional Chinese medicine composition according to claim 1, characterized in that, It is made from 40 parts by weight of sweet flag, 40 parts of iron rod, 60 parts of benzoin, and 100 parts of catechu.

3. The traditional Chinese medicine composition according to claim 1, characterized in that, The reflux extraction in steps S1, S2 and S4 is performed 2 to 3 times, with a duration of 0.8 to 1.2 hours per extraction.

4. The traditional Chinese medicine composition according to claim 1, characterized in that, The static extraction in step S3 is performed 2 to 3 times, with a duration of 22 to 26 hours per extraction.

5. The traditional Chinese medicine composition according to claim 1, characterized in that, The pulverization described in steps S1 to S4 is pulverization into the coarsest powder, passing through 10 mesh.

6. Use of the traditional Chinese medicine composition according to any one of claims 1 to 5 in the preparation of a medicament for treating hyperuricemia.

7. The use according to claim 6, characterized in that, The medicine includes one of the following: compound preparation, ointment, granule, pill, capsule, syrup, spray, powder, and tablet.

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