A pharmaceutical composition for preventing and treating hyperuricemia, accompanying liver and kidney injury and combined gouty arthritis, and a preparation method and application thereof

Abstract

The application provides a kind of for preventing and treating hyperuricemia, accompanying liver and kidney damage and combined gouty arthritis pharmaceutical composition and its preparation method and application, belong to biological medicine technical field.A kind of pharmaceutical composition, including the following mass parts of traditional Chinese medicine raw materials are made: 10~60 parts of radix puerariae, 5~20 parts of atractylodes, 5~20 parts of phellodendri cortex, 5~20 parts of smilax glabra and 5~20 parts of radix astragali.The pharmaceutical composition obtained by using the optimized preparation process, not only can quickly and continuously reduce blood uric acid level, but also can regulate metabolic disorder, control systemic inflammation, protect liver and kidney function, relieve acute phase arthritis inflammation swelling, can realize clinical treatment hyperuricemia, accompanying liver and kidney damage and combined acute gouty arthritis.

Description

Technical Field

[0001] This application belongs to the field of biopharmaceutical technology, specifically relating to a pharmaceutical composition for the prevention and treatment of hyperuricemia, associated liver and kidney damage and gouty arthritis, as well as its preparation method and application. Background Technology

[0002] Hyperuricemia (HUA) is a high-risk factor for many chronic diseases, including chronic kidney disease, hypertension, cardiovascular and cerebrovascular diseases, and diabetes. It is a systemic disease affecting multiple systems and has become the most common systemic metabolic disease after diabetes. Serum uric acid (SUC) is a typical symptom of HUA. Based on the cause of elevated SUC, HUA can be classified into three types: impaired uric acid excretion, excessive uric acid production, and mixed types. Endogenous uric acid metabolism disorders or excretion impairments are the main causes of HUA, while increased exogenous uric acid metabolism is a secondary cause. The kidneys are the key organs for uric acid excretion in the body; therefore, uric acid metabolism disorders or excretion impairments are often accompanied by kidney damage.

[0003] Currently, clinical medications for treating hyperuricemia include xanthine oxidase inhibitors that inhibit uric acid synthesis, such as allopurinol and febuxostat; uricosuric drugs, such as benzbromarone; and recombinant uricase preparations, such as polyethylene glycol recombinant uricase. However, long-term use of Western medicine for treatment has significant adverse reactions such as liver and kidney damage, gastrointestinal reactions, and cardiovascular risks. Furthermore, there is a contradiction between medication use during the acute and remission phases, and serum uric acid levels tend to rebound after discontinuation of medication, resulting in poor long-term patient compliance. Summary of the Invention

[0004] The purpose of this invention is to provide a pharmaceutical composition for the prevention and treatment of hyperuricemia, associated liver and kidney damage, and gouty arthritis. This composition can not only rapidly and continuously reduce blood uric acid levels, but also regulate metabolic disorders, control systemic inflammation, protect liver and kidney function, and relieve inflammation and swelling of joints in the acute phase. It can achieve clinical treatment of hyperuricemia, associated liver and kidney damage, and gouty arthritis.

[0005] This invention provides a pharmaceutical composition comprising the following parts by weight of traditional Chinese medicine raw materials: 10-60 parts of kudzu root, 5-20 parts of atractylodes rhizome, 5-20 parts of phellodendron bark, 5-20 parts of smilax glabra rhizome, and 5-20 parts of achyranthes root.

[0006] Preferably, it is made from the following Chinese medicinal materials in parts by weight: 20-40 parts of kudzu root, 7-15 parts of atractylodes rhizome, 8-13 parts of phellodendron bark, 10-17 parts of smilax glabra rhizome, and 6-15 parts of achyranthes bidentata root.

[0007] This invention provides a method for preparing the pharmaceutical composition, comprising the following steps: Phellodendron bark and Smilax glabra were extracted separately or in combination by decoction to obtain a decoction. Atractylodes lancea was extracted by steam distillation to obtain Atractylodes lancea volatile oil, Atractylodes lancea water extract, and Atractylodes lancea residue; The Atractylodes lancea residue, Pueraria lobata and Achyranthes bidentata were extracted with alcohol, either separately or in combination, to obtain an alcohol extract. The decoction and Atractylodes lancea water extract were dehydrated to obtain the water extract; The volatile oil of Atractylodes lancea was encapsulated and then mixed with alcoholic and water extracts to obtain a pharmaceutical composition.

[0008] Preferably, the extraction of Phellodendron bark and Smilax glabra is performed 2 to 3 times; the mass ratio of the total mass of Phellodendron bark and Smilax glabra to water is 1 to 6 to 8 during each extraction; and the extraction time is 1.5 to 2 hours.

[0009] Preferably, during the steam distillation extraction, the mass ratio of Atractylodes lancea to water is 1:7.5~8.5; and the extraction time is 5.5~6.5 hours. The method for encapsulating the volatile oil of Atractylodes lancea involves adding the volatile oil of Atractylodes lancea to an encapsulating excipient or an encapsulating excipient solution and encapsulating it using at least one of the following methods: colloid milling, ultrasonication, grinding, and water-saturated solution method.

[0010] Preferably, the volume concentration of the alcohol during the alcohol extraction is 68%~72%; the alcohol includes ethanol; the alcohol extraction is performed 2~3 times; and the extraction time for each extraction is 1.5~2 hours.

[0011] This invention provides a drug for preventing and treating hyperuricemia complicated with gouty arthritis, comprising the drug composition or the drug composition obtained by the preparation method and pharmaceutically acceptable excipients.

[0012] Preferably, the pharmaceutically acceptable excipients include fillers and flavoring agents; The filler includes dextrin; the filler accounts for 10% to 50% of the total mass of the formulation; The flavoring agent comprises stevioside and citric acid; the stevioside accounts for 0.8% to 1.2% of the mass of the pharmaceutical composition; and the citric acid accounts for 0.4% to 0.6% of the mass of the pharmaceutical composition.

[0013] The present invention provides the use of the pharmaceutical composition or the pharmaceutical composition obtained by the preparation method in the preparation of a medicament for the prevention and / or treatment of at least one of the following diseases: hyperuricemia, hyperuricemia with liver and kidney damage, and hyperuricemia complicated with gouty arthritis.

[0014] Preferably, the complications of hyperuricemia combined with gouty arthritis also include kidney and / or liver damage.

[0015] This invention provides a pharmaceutical composition comprising the following parts by weight of traditional Chinese medicine raw materials: 10-60 parts of kudzu root, 5-20 parts of atractylodes rhizome, 5-20 parts of phellodendron bark, 5-20 parts of smilax glabra rhizome, and 5-20 parts of achyranthes root. The pharmaceutical composition is based on the principles of "strengthening the spleen and kidneys, clearing heat and dampness, and resolving turbidity and unblocking the meridians." It treats hyperuricemia by improving blood uric acid levels, regulating lipid metabolism disorders, and reducing local and systemic inflammatory responses. It has a significant protective effect against liver and kidney damage caused by hyperuricemia and can also prevent and alleviate acute gout attacks.

[0016] In the present invention, adenine plus yeast induction as well as potassium oxonate plus yeast induction are adopted to establish a rat hyperuricemia model, combined with intra-articular ankle injection to induce acute arthritis injury, so as to successfully construct an animal model highly consistent with the pathogenesis of clinical hyperuricemia combined with acute gouty arthritis, which is used to evaluate the efficacy of the Gezhu pharmaceutical composition. The experimental results show that the serum uric acid of the model rats is significantly increased. After 2 weeks and 4 weeks of intervention with the Gezhu pharmaceutical composition, the serum uric acid level of the rats is progressively and significantly reduced, and the uric acid-lowering effect is more stable after long-term administration, suggesting that the Gezhu pharmaceutical composition can not only rapidly reduce the serum uric acid concentration, but also maintain the stability of uric acid metabolism level for a long time. C-reactive protein (CRP) is one of the markers of acute inflammation and tissue injury in the body, and homocysteine (Hcy) is a predictor of systemic chronic low-grade inflammation, vascular endothelial injury, etc. The elevation of both levels is highly correlated with hyperuricemia and gout attacks. The experimental results show that the serum CRP and Hcy levels of the model rats are significantly increased, suggesting that hyperuricemia can induce the activation of systemic inflammatory response. After treatment with the Gezhu pharmaceutical composition, the levels of CRP and Hcy are both significantly reduced, suggesting that the Gezhu pharmaceutical composition can inhibit the systemic inflammatory response, not only can improve local joint inflammation and reduce inflammation-mediated tissue and vascular damage. Compared with the simple uric acid-lowering treatment of Western medicine, the Gezhu pharmaceutical composition has the characteristics of local anti-inflammation and overall regulation, and has more obvious clinical treatment advantages. At the same time, both clinical epidemiology and basic research have confirmed that hyperuricemia and gout are metabolism-related diseases, and patients often have multiple systemic lesions such as dyslipidemia, glucose metabolism disorder, and renal function injury, while abnormal lipid and glucose metabolism and decreased renal function will further aggravate uric acid excretion disorder, forming a vicious cycle of "uric acid elevation - metabolic disorder - target organ injury". The results of this experiment show that the levels of serum triglyceride (TG), total cholesterol (TC), blood glucose (GLU), aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatinine (CRE) and urea (UREA) in the model rats are all significantly increased; after treatment with the Gezhu pharmaceutical composition, all the above indicators are significantly reduced, suggesting that the Gezhu pharmaceutical composition has a therapeutic effect on improving lipid metabolism disorder. A particularly obvious manifestation is that the liver and kidney functions of the rats in the model group are significantly impaired, the contents of AST, ALT, CRE and UREA are significantly increased, and the liver and kidney functions are significantly improved after administration of the Gezhu pharmaceutical composition, with the contents of AST, ALT, CRE and UREA significantly reduced, which solves the dual problems of hyperuricemia and liver and kidney damage caused by existing chemical drugs, and meets the major clinical needs.

[0017] Meanwhile, the core pathogenesis of acute gouty arthritis is that sodium urate crystals deposit in the joint cavity, activate the innate immune response, induce neutrophil infiltration and massive release of inflammatory factors, and cause local joint hyperemia, edema and inflammatory exudation. The results of this experiment show that the ankle joint volume of model rats increases significantly, and the ankle joint volume decreases significantly after treatment with the Gezhu pharmaceutical composition, indicating that the Gezhu pharmaceutical composition can directly act on the inflammatory lesion site, effectively inhibit the local acute inflammatory reaction induced by sodium urate crystals, reduce joint soft tissue edema and inflammatory infiltration, and rapidly relieve the joint swelling symptom during the acute attack of gout, which is consistent with the basic therapeutic principle of "clearing heat and promoting diuresis, detumescence and analgesia, unblocking collaterals and resolving masses" of traditional Chinese medicine for the treatment of acute gout stage, and reflects the unique advantages of traditional Chinese medicine compound prescriptions in the symptomatic treatment of acute gout stage.

[0018] In summary, targeting multiple pathological injury links of hyperuricemia, accompanying liver and kidney damage, and complicated acute gouty arthritis, the Gezhu pharmaceutical composition can not only rapidly and continuously reduce the blood uric acid level, control systemic inflammation, regulate metabolic disorders and protect renal function, but also relieve joint inflammation and swelling in the acute stage. This provides a preclinical experimental basis for the clinical application of the Gezhu pharmaceutical composition in the treatment of hyperuricemia, gouty arthritis and related metabolic complications, and provides new ideas and methods for the prevention and treatment of metabolic diseases with traditional Chinese medicine.

[0019] The present invention provides a preparation method of the pharmaceutical composition, wherein Phellodendri Chinensis Cortex and Smilacis Glabrae Rhizoma are extracted separately or in combination by a water decoction method to obtain a water decoction solution; Atractylodis Rhizoma is extracted by a steam distillation method to obtain Atractylodis Rhizoma volatile oil, Atractylodis Rhizoma water extract and Atractylodis Rhizoma medicinal residues; the Atractylodis Rhizoma medicinal residues, Puerariae Lobatae Radix and Achyranthis Bidentatae Radix are subjected to alcohol extraction separately or in combination to obtain an alcohol extract; the water decoction solution and the Atractylodis Rhizoma water extract are dehydrated to obtain a water extract; the Atractylodis Rhizoma volatile oil is subjected to inclusion treatment and then mixed with the alcohol extract and the water extract to obtain the pharmaceutical composition. Based on the formula of the pharmaceutical composition, the present invention takes the contents of puerarin in Puerariae Lobatae Radix, atractylodin and atractylon (volatile oil) in Atractylodis Rhizoma, berberine in Phellodendri Chinensis Cortex, astilbin in Smilacis Glabrae Rhizoma, and β-ecdysterone in Achyranthis Bidentatae Radix as investigation indexes, and screens out a set of preparation processes that can efficiently extract active ingredients, which has obvious advantages in reducing the blood uric acid level in hyperuricemia, and has good therapeutic effects on related metabolic complications and gouty arthritis. The preparation method provided by the present invention is simple to operate, can efficiently extract active ingredients with the effects of treating hyperuricemia, preventing metabolic disorders and complicated gouty arthritis, and has great popularization and application value.

[0020] The extraction process of the present invention has the following characteristics: 1. Phellodendri Chinensis Cortex is extracted with water to maximize the extraction of active ingredients The main active ingredients of Phellodendron bark are alkaloid salts such as berberine and berberine. Their solubility in hot water is acceptable, but their solubility in high-concentration ethanol decreases (especially in sulfate or hydrochloride forms). Phellodendron bark also contains a large amount of mucilage, resin, and pigments.

[0021] Process advantages: If Phellodendron bark is used in alcohol extraction, a large amount of these resins and pigments will dissolve, which not only increases the viscosity of the alcohol extract and the difficulty of purification, but also leads to an excessively bitter taste and dark color in the final preparation. Water extraction, on the other hand, can effectively extract alkaloids and actively avoid a large number of alcohol-soluble impurities, ensuring the purity of the subsequent alcohol extraction stage.

[0022] 2. Based on the characteristics of the main active ingredients of the medicinal materials, water extraction and alcohol extraction were carried out respectively. To maximize the extraction of the effective components for treating hyperuricemia from this formula, the preparation process clearly divides the medicinal materials into a water extraction group (Phellodendron bark + Smilax glabra) and an alcohol extraction group (Atractylodes lancea residue + Pueraria lobata + Achyranthes bidentata) based on the polarity differences of the main active ingredients in different medicinal materials.

[0023] Water extraction group (Phellodendron bark + Smilax glabra): Astilbene in Smilax glabra and alkaloid salts in Phellodendron bark are both moderately polar and have good water solubility. Water extraction can fully dissolve them, and it is low-cost and safe to operate.

[0024] Alcohol-extracted group (Atractylodes lancea residue + Pueraria lobata + Achyranthes bidentata): Puerarin in Pueraria lobata, ecdysterone in Achyranthes bidentata, and atractylodes lancea residue, along with other non-volatile components, are all alcohol-soluble. If water extraction is used instead, the extraction rate will be extremely low (the dissolution rate of puerarin in water is less than 1 / 3 of that in alcohol extraction).

[0025] The above-mentioned preparation method, which involves separate extraction with water and alcohol, ensures that the "core active ingredients" of each medicinal material can be extracted to the maximum extent.

[0026] 3. Comprehensive coverage of the medicinal components of Atractylodes lancea The process first involves steam distillation of Atractylodes lancea to extract its volatile oil components; the residue is then subjected to alcohol extraction to simultaneously obtain the non-volatile components. Through this sequential extraction process, a comprehensive and in-depth utilization of the active ingredients of Atractylodes lancea is achieved.

[0027] (1) Distillation: The volatile oil is extracted by steam distillation, and its volatile active components, such as atractylodes lactone and atractylone, are extracted to be responsible for anti-inflammatory and regulating intestinal uric acid excretion.

[0028] (2) Inclusion: β-cyclodextrin inclusion is used to solve the problems of easy oxidation of volatile oil, irritation of gastric mucosa, and stability.

[0029] (3) Alcohol extraction of herb residues: The herb residues after volatile oil extraction of Atractylodis Rhizoma are still rich in non-volatile components such as atractylodin and polysaccharides, which just have potential uric acid-lowering effects. Mixing them with Puerariae Lobatae Radix and Achyranthis Bidentatae Radix for alcohol extraction not only acts as a "filler" to adsorb pigments in the alcohol extract, but also realizes the full utilization of the active pharmaceutical components of Atractylodis Rhizoma.

[0030] The described preparation method does not blindly pursue alcohol extraction for all medicinal materials or water extraction for all medicinal materials. Instead, based on the chemical properties that phellodendrine is intolerant to high-concentration alcohol and puerarin is insoluble in water, the formula is split into water extraction (Phellodendri Chinensis Cortex + Smilacis Glabrae Rhizoma) and alcohol extraction (Puerariae Lobatae Radix + Achyranthis Bidentatae Radix + Atractylodis Rhizoma herb residues), and the shortcoming of volatile oil is compensated by an inclusion technology; this preparation method embodies the technical idea of modern traditional Chinese medicine preparations transitioning from "empirical decoction" to "component-structure-based pharmaceutical manufacturing". Brief Description of the Drawings

[0031] Figure 1 is a first flow chart of the preparation process of the Gezhu pharmaceutical composition; Figure 2 is a second flow chart of the preparation process of the Gezhu pharmaceutical composition; Figure 3 is a third flow chart of the preparation process of the Gezhu pharmaceutical composition; Figure 4 is a fourth flow chart of the preparation process of the Gezhu pharmaceutical composition; Figure 5 shows the renal HE staining results of each group of mice in Example 4; Figure 6 shows the hepatic HE staining results of each group of mice in Example 4; Figure 7 shows the ankle joint HE staining results of each group of mice in Example 5; Figure 8 shows the renal HE staining results of each group of mice in Example 7; Figure 9 shows the hepatic HE staining results of each group of mice in Example 7; Figure 10 shows the ankle joint HE staining results of each group of mice in Example 8. Detailed Description of the Embodiments

[0032] The present invention provides a pharmaceutical composition, which is prepared from the following traditional Chinese medicinal raw materials in parts by mass: 10-60 parts of Puerariae Lobatae Radix, 5-20 parts of Atractylodis Rhizoma, 5-20 parts of Phellodendri Chinensis Cortex, 5-20 parts of Smilacis Glabrae Rhizoma, and 5-20 parts of Achyranthis Bidentatae Radix.

[0033] In the present invention, the pharmaceutical composition is preferably prepared from the following traditional Chinese medicinal raw materials in parts by mass: 20-40 parts of Puerariae Lobatae Radix, 7-15 parts of Atractylodis Rhizoma, 8-13 parts of Phellodendri Chinensis Cortex, 10-17 parts of Smilacis Glabrae Rhizoma, and 6-15 parts of Achyranthis Bidentatae Radix; it may be prepared from the following traditional Chinese medicinal raw materials in parts by mass: 30 parts of Puerariae Lobatae Radix, 10 parts of Atractylodis Rhizoma, 10 parts of Phellodendri Chinensis Cortex, 15 parts of Smilacis Glabrae Rhizoma, and 6 parts of Achyranthis Bidentatae Radix.

[0034] In this invention, the kudzu root has the functions of dispelling wind and dampness, promoting blood circulation and regulating menstruation, and invigorating the spleen with its aromatic properties. The atractylodes rhizome is pungent, bitter, and warm in nature, and enters the spleen, stomach, and liver meridians; it has the functions of drying dampness and strengthening the spleen, dispelling wind and cold, and improving eyesight. The phellodendron bark is bitter and cold in nature; it enters the kidney and bladder meridians, and has the functions of clearing heat and drying dampness, purging fire and eliminating steaming heat, and detoxifying and healing sores. The smilax rhizome is sweet, bland, and neutral in nature; it enters the liver and stomach meridians; it has the functions of detoxifying, eliminating dampness, and promoting joint mobility. The achyranthes root is bitter, sweet, and sour in nature, and neutral in nature; it enters the liver and kidney meridians; it has the functions of removing blood stasis and regulating menstruation, tonifying the liver and kidneys, strengthening tendons and bones, promoting urination and relieving strangury, and guiding blood downwards. The achyranthes root has good antioxidant properties. This invention does not have special restrictions on the sources of the phellodendron bark, smilax rhizome, atractylodes rhizome, kudzu root, and achyranthes root; conventional sources of medicinal materials well known in the art are acceptable.

[0035] This invention provides a method for preparing the pharmaceutical composition, comprising the following steps: Phellodendron bark and Smilax glabra were extracted separately or in combination by decoction to obtain a decoction. Atractylodes lancea was extracted by steam distillation to obtain Atractylodes lancea volatile oil, Atractylodes lancea water extract, and Atractylodes lancea residue; The Atractylodes lancea residue, Pueraria lobata and Achyranthes bidentata were extracted with alcohol, either separately or in combination, to obtain an alcohol extract. The decoction and Atractylodes lancea water extract were dehydrated to obtain the water extract; The volatile oil of Atractylodes lancea was encapsulated and then mixed with alcoholic and water extracts to obtain a pharmaceutical composition.

[0036] Based on the raw material composition of the pharmaceutical composition, the present invention uses the whole formula decoction method (process 1), whole formula alcohol extraction method (process 2), water decoction combined with alcohol extraction method (process 3), and water extraction and alcohol precipitation combined with alcohol extraction method (process 4) to extract the active ingredients of the drug. The blood uric acid level was used as the evaluation index. The results showed that the water decoction method combined with alcohol extraction method (process 3) had the best effect.

[0037] In this invention, Phellodendron bark and Smilax glabra are extracted separately or in combination by water decoction to obtain a decoction.

[0038] In this invention, the content of berberine in Phellodendron bark and astilbin in Smilax glabra rhizome are used as evaluation indicators to optimize the extraction process parameters. The extraction of Phellodendron bark and Smilax glabra rhizome is preferably performed 2-3 times, or 2 times. The mass ratio of the total mass of Phellodendron bark and Smilax glabra rhizome to water is preferably 1-6-8 for each extraction. For the first extraction, the mass ratio is preferably 1:8; for the second or third extraction, the mass ratio is preferably 1:6. This invention does not have any special restrictions on the type of water used; pharmaceutical water well-known in the art can be used, such as drinking water, purified water, distilled water, or water for injection. The extraction time for each extraction is preferably 1.5-2 hours, or 1.8-2 hours.

[0039] In one embodiment of the present invention, the number of extractions (1, 2, or 3 times), extraction time (1 h, 1.5 h, and 2 h), and solvent volume (6 times, 8 times, and 10 times) for the decoction of Phellodendron bark and Smilax glabra were optimized. The results showed that adding 8 times the amount of water during the first extraction, extracting 3 times, and 2 h each time, was the optimal process. Furthermore, the transfer rates of the active ingredients (berberine and astilbin) were comparable between extractions of 2 and 3 times, with no significant difference observed. To improve extraction efficiency and reduce costs, the optimal number of extractions was 2. The drug yields of Phellodendron bark and Smilax glabra under the optimal extraction process were 17.16%–17.70%, indicating that the decoction method is stable and feasible.

[0040] This invention uses steam distillation to extract Atractylodes lancea to obtain Atractylodes lancea volatile oil, Atractylodes lancea water extract, and Atractylodes lancea residue.

[0041] In this invention, during the steam distillation extraction, the preferred mass ratio of Atractylodes lancea to water is 1:7.5~8.5, and can be 1:8. The preferred extraction time is 5.5~6.5 hours, and can be 6 hours. During extraction, the Atractylodes lancea is preferably pulverized to coarse powder, i.e., the coarse powder particle size is such that all particles pass through a No. 2 sieve: sieve aperture inner diameter 850 μm (±29 μm), approximately 24 mesh, with ≤ 40% of the powder passing through a No. 4 sieve (250 μm).

[0042] In one embodiment of the present invention, the parameters of steam distillation extraction of Atractylodes lancea were optimized using the oil yield as an indicator. By optimizing the water addition ratio and extraction time, the results showed that using 8 times the amount of water and extracting for 6 hours was beneficial to obtaining the highest oil yield and maintaining a high extraction efficiency.

[0043] After obtaining the Atractylodes lancea residue, the present invention performs alcohol extraction on the Atractylodes lancea residue, Pueraria lobata and Achyranthes bidentata separately or in combination to obtain an alcohol extract.

[0044] In this invention, the volume concentration of the alcohol during the alcohol extraction is preferably 68%~72%, and can be 70%. The alcohol preferably includes ethanol. The solvent for the ethanol is preferably water. The number of alcohol extractions is preferably 2~3 times, and can be 2 times. The extraction time for each extraction is preferably 1.5~2 hours, and can be 2 hours. The volume ratio of the alcohol to the total mass of the Atractylodes lancea residue, Pueraria lobata, and Achyranthes bidentata is preferably 1:8~10, and can be 1:9. The alcohol extraction is preferably a reflux extraction using an aqueous ethanol solution.

[0045] In one embodiment of the present invention, the contents of atractylodes in Atractylodes lancea, berberine in Phellodendron chinense, astilbin in Smilax glabra, and β-ecdysterone in Achyranthes bidentata were used as evaluation indicators. The effects of alcohol extraction were evaluated by optimizing the number of extractions (1, 2, and 3 times), extraction time (1 h, 1.5 h, and 2 h), solvent volume (6, 8, and 10 times), and solvent concentration (50%, 60%, and 70%). The results showed that the optimal process was to use 10 times the solvent for the first extraction, with the solvent being 70% ethanol, and to extract twice, with each extraction lasting 2 hours.

[0046] In this invention, the alcohol extraction process preferably includes ethanol recovery and concentration to obtain an alcohol-extracted thick paste. The method for ethanol recovery and concentration is preferably vacuum concentration to collect the ethanol. The vacuum concentration temperature is preferably not higher than 80°C. The concentrated paste has a density of 1.30~1.35.

[0047] The present invention removes water from the decoction and Atractylodes lancea water extract to obtain the water extract.

[0048] In this invention, the method for removing moisture preferably includes concentration. The concentration method is preferably vacuum concentration. The temperature of the vacuum concentration is preferably not higher than 80°C. The concentration is carried out to a water-extracted thick paste with a density of 1.30~1.35.

[0049] After obtaining the aqueous extract, the present invention combines the Atractylodes lancea volatile oil with the alcohol extract and aqueous extract to obtain a pharmaceutical composition.

[0050] In this invention, the method for encapsulating Atractylodes lancea volatile oil involves adding the volatile oil to an encapsulating excipient or an encapsulating excipient solution and performing encapsulation treatment using at least one of the following methods: colloid milling, ultrasonication, grinding, and water-saturated solution method. The encapsulating excipient is preferably β-cyclodextrin. When grinding is used, the encapsulating excipient exists in solid form; when colloid milling, ultrasonication, or water-saturated solution method is used, the encapsulating excipient is in solution form. The concentration of the encapsulating excipient solution is preferably 16%~34%, and can also be 20%~32%. Encapsulation treatment of the Atractylodes lancea volatile oil improves stability, reduces volatility, minimizes loss, improves physical properties, facilitates formulation, solubilizes, and masks unpleasant / irritating odors.

[0051] In this embodiment of the invention, the inclusion rates of different methods were measured and evaluated. The inclusion rates of the water-saturated solution method, grinding, and ultrasonic methods were all higher than those of the colloid mill, with the water-saturated solution method, grinding, and ultrasonic methods all showing higher inclusion rates than the colloid mill. After being placed under different conditions for 10 days, the main index components of the four inclusion methods all decreased. Among them, the water-saturated solution method and ultrasonic method showed a slower decrease in index components and higher retention rates. From a practical production perspective, the water-saturated solution method is preferred.

[0052] In this invention, the mixing process preferably further includes drying, pulverizing, and sieving. The drying method preferably includes vacuum drying or freeze drying. The sieving is preferably performed through an 80-mesh sieve, and the undersize material is collected to obtain a dry paste powder.

[0053] This invention provides a drug for preventing and treating hyperuricemia complicated with gouty arthritis, comprising the drug composition or the drug composition obtained by the preparation method and pharmaceutically acceptable excipients.

[0054] In this invention, the preferred dosage form of the drug is an oral dosage form such as powder, granule, or capsule. In this embodiment, the preparation process of the dosage form is illustrated using granules as an example. The preferred method for preparing the granules is wet granulation.

[0055] In this invention, the pharmaceutically acceptable excipients preferably include fillers and flavoring agents. The fillers preferably include dextrin. The fillers preferably constitute 10% to 50% of the pharmaceutical composition by mass, but can be 20% to 40%, 25% to 35%, or 30%. Example results show that dextrin has better formability than lactose; the formability of dextrin as a filler reaches 97.67%, while the formability of kudzu granules prepared with the same addition ratio of lactose is 59.05%.

[0056] In this invention, the flavoring agent preferably includes stevioside and citric acid. The stevioside accounts for 0.8% to 1.2% of the mass of the pharmaceutical composition, and can be 1.0%. The citric acid accounts for 0.4% to 0.6% of the mass of the pharmaceutical composition, and can be 0.5%. The addition of stevioside and citric acid in the above proportions can effectively improve the bitter taste of the original medicine, enhance its sweet and sour flavor, and make it widely acceptable to a broad audience.

[0057] The present invention provides the use of the pharmaceutical composition or the pharmaceutical composition obtained by the preparation method in the preparation of a medicament for the prevention and / or treatment of at least one of the following diseases: hyperuricemia, associated liver and kidney damage, and gouty arthritis.

[0058] In this invention, the prevention and / or treatment of hyperuricemia preferably involves lowering serum uric acid levels. Liver and kidney damage associated with hyperuricemia includes lowering serum uric acid levels, regulating lipid and glucose metabolism disorders, improving liver and kidney function, and inhibiting local and systemic inflammatory responses. Hyperuricemia complicated with gouty arthritis includes lowering serum uric acid levels and inhibiting local and systemic inflammatory responses. The prevention and / or treatment of gouty arthritis preferably involves reducing joint volume, acting directly on the inflammatory lesion site, effectively inhibiting the local acute inflammatory response induced by sodium urate crystals, reducing joint soft tissue edema and inflammatory infiltration, and rapidly relieving joint swelling symptoms during acute gout attacks. This aligns with the basic principles of traditional Chinese medicine for treating acute gout attacks: "clearing heat and dampness, reducing swelling and relieving pain, and unblocking meridians and dispersing nodules," demonstrating the unique advantages of the pharmaceutical composition provided by this invention in symptomatic treatment of acute gout attacks. The kudzu root pharmaceutical composition has the effects of reducing joint vascular proliferation, reducing bone destruction, reducing synovial cell proliferation and inflammatory cell infiltration, and has a significant protective effect against gouty arthritis.

[0059] In this embodiment of the invention, adenine and yeast induction and potassium oxonate and yeast induction, combined with intra-articular injection of the ankle joint, are used to induce acute gouty arthritis. Combined with the pathological manifestations of increased uric acid production, excretion disorders, acute joint inflammation, and systemic metabolic disorders in clinical practice, an animal model that closely matches the pathogenesis of clinical hyperuricemia complicated with acute gouty arthritis is used to treat the pharmacological effects of the kudzu root drug composition in rats with hyperuricemia complicated with acute gouty arthritis.

[0060] In the embodiments of the present invention, the efficacy of the pharmaceutical composition is evaluated based on clinical indicators (serum uric acid level, CRP, Hcy, TG, TC, AST, ALT, CRE, UREA and GLU levels) and joint pathological test results. The results show that the serum uric acid level of the model rats is significantly increased after modeling, and the serum uric acid level is significantly decreased in the three dose groups of the positive drugs benzbromarone, Tongfengshu, and Gezhu Granule after 2 weeks and 4 weeks of administration, with significant differences compared with the model control group. The same changes are presented in the test results of CRP, Hcy, TG, TC, AST, ALT, CRE, UREA and GLU. Regarding the joint pathological test results, the ankle joint volume of the model rats is significantly increased, while the three dose groups of the positive control drugs benzbromarone, Tongfengshu, and Gezhu Granule can all significantly reduce the volume of the right ankle joint of the rats after modeling, with significant differences compared with the model control group. Meanwhile, the results of light microscopy examination (HE staining results) show that the synovial cells of the joints of the rats in the model control group have mild to severe hyperplasia; the interstitium is congested and edematous, accompanied by infiltration of neutrophils, lymphocytes and monocytes, fibrinous exudation, narrowed joint space, and a large amount of inflammatory tissue necrosis can be seen; the small blood vessels are dilated and proliferated, pannus is formed, and the bone tissue is destroyed in a moth-eaten pattern; mild to severe inflammatory cell infiltration and fibrous tissue hyperplasia are observed around the joint, with significant differences compared with the normal control group. In the high-dose group of the Gezhu pharmaceutical composition, the joint blood vessels of the rats have slight or mild hyperplasia, and the bone destruction is significantly reduced, with significant differences compared with the model control group; the degree of synovial cell hyperplasia and inflammatory cell infiltration shows a decreasing trend. It indicates that the Gezhu pharmaceutical composition has a significant protective effect on gouty arthritis.

[0061] In the present invention, since hyperuricemia and gouty arthritis are metabolism-related diseases, patients often have multiple systemic lesions such as dyslipidemia, glucose metabolism disorder, and renal function impairment, while abnormal lipid and glucose metabolism and decreased renal function will further aggravate uric acid excretion disorder, forming a vicious cycle of "elevated uric acid - metabolic disorder - target organ damage". Therefore, the complications of hyperuricemia combined with gouty arthritis preferably further include kidney and / or liver damage. The Gezhu pharmaceutical composition has a significant protective effect on liver and kidney damage caused by hyperuricemia. It can significantly reduce the damage to glomeruli and renal tubules in rats, and reduce the scope and degree of renal interstitial fibrosis and inflammatory cell infiltration; it can reduce the scope and degree of hepatocyte vacuolization.

[0062] In this embodiment of the invention, the results of renal pathological examination showed that in the model control group, rats exhibited mild to severe proliferation of the glomerular mesangial matrix and partial glomerular sclerosis; proximal tubular epithelial cells showed degeneration / necrosis and atrophy, with thickened basement membrane and protein casts and urate crystals observed in some tubular lumens; the renal interstitium showed mild to moderate fibrosis and multifocal inflammatory cell infiltration, predominantly mononuclear cells, which was significantly different from the normal control group. Compared with the model group, the renal changes in each treatment group were alleviated to varying degrees. Specifically, the benzbromarone group and the high, medium, and low dose groups of the kudzu root drug combination showed significantly reduced glomerular mesangial matrix deposition; a significantly reduced range of renal tubular degeneration / necrosis; and a significantly reduced range and degree of interstitial fibrosis and inflammatory cell infiltration, which were significantly different from the model control group, suggesting a significant protective effect against renal injury. In the Gout Relief Group, the extent and degree of glomerular damage, renal interstitial fibrosis, and inflammatory cell infiltration in rats were significantly reduced, showing a significant difference from the model control group; renal tubules showed mild to moderate degeneration / necrosis, with a decreasing score trend.

[0063] In this embodiment of the invention, liver pathological examination results showed that the hepatocytes of the model control group rats were swollen and degenerated, with mild to moderate cytoplasmic vacuolation, which was significantly different from that of the normal control group. The extent and degree of hepatocyte vacuolation in the benzbromarone group and the high, medium, and low dose groups of the kudzu root drug combination were significantly reduced, showing a significant difference from the model control group, suggesting a significant protective effect against liver injury. The hepatocyte vacuolation score of the gout relief group rats showed a decreasing trend.

[0064] The following detailed description, in conjunction with embodiments, illustrates a pharmaceutical composition for the prevention and treatment of hyperuricemia, accompanied by liver and kidney damage, and complicated by gouty arthritis, as well as its preparation method and application. However, these descriptions should not be construed as limiting the scope of protection of this invention.

[0065] Example 1 Screening test of the optimal extraction process route for the kudzu root drug composition The intended clinical indication for Geshu granules is the treatment of hyperuricemia. This embodiment provides a basis for determining the optimal preparation process through efficacy screening. In the experiment, a rat model of hyperuricemia was established using potassium cyanide and yeast feed. Samples from different processes were administered by gavage at two dosage groups: 6.6 g crude drug / kg / d and 3.3 g crude drug / kg / d (equivalent to one-times and one-half-times of the human clinical dose, respectively), once daily for 14 consecutive days.

[0066] The Ge-Shu medicinal composition comprises five Chinese medicinal herbs: 10 parts by weight of kudzu root, 5 parts by weight of atractylodes rhizome, 5 parts by weight of phellodendron bark, 5 parts by weight of smilax glabra rhizome, and 5 parts by weight of achyranthes root. Based on the active ingredients of the herbs in the prescription, four extraction processes were designed, as detailed below. Figures 1-4 .

[0067] 1 Test Materials 1.1 Gezhu Granules (Dry Extract Powder) There are 4 samples of Gezhu Granules (Dry Extract Powder), labeled as Sample 1#, 2#, 3# and 4# respectively. Properties: dry traditional Chinese medicine dry extract powder; Characteristics: yellow to brown solid powder; The contents are 2.80 g crude drug / g, 5.75 g crude drug / g, 3.98 g crude drug / g and 5.85 g crude drug / g respectively. In the test, two dose groups of 6.6 g crude drug / kg / d and 3.3 g crude drug / kg / d were set for rats, which are equivalent to the equal dose and 1 / 2 of the clinical dose for human respectively.

[0068] 1.2 Positive Control Drug Benzbromarone Tablets, a product of Hermes Arzneimittel GmbH, Germany. Clinical indications: primary hyperuricemia, intermission of gouty arthritis, gouty tophus, etc.; National Medical Product Approval No. J20180056; Production date: 20220221, Expiry date: 20270220; Specification: 50mg / tablet; Characteristics: the product is white or off-white tablet; Usage and dosage: for adults, oral administration of 50mg (1 tablet) each time, once a day 1.3 Test reagents are shown in Table 1 Table 1

[0069] 1.4 Test Animals 120 male SD rats, SPF grade, weighing 180~220g, were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. Animal License No.: SCXK (Beijing) 2021-0006, Certificate No.: No.110011241104310968.

[0070] 1.5 Test instruments are shown in Table 2.

[0071] Table 2

[0072] 2 Test Methods 2.1 Dose Design and Drug Preparation 2.1.1 Test Drug Table 3 Test Drug and Preparation Method

[0073] 2.1.2 Positive Control Drug For Benzbromarone Tablets, the clinical dose for human is 50mg / 60kg / d, that is 0.83mg / kg / d; the dosage for rats in this test is 4.65mg / kg / d, which is equivalent to the equal clinical dose for human. The drug is administered by gavage at 1ml / 100g body weight. 2.2 Test Grouping and Administration One hundred and twenty male SD rats (SPF grade, weighing 200±20g) were used. After 3 days of acclimatization, their baseline uric acid levels were measured. One hundred of these rats with uric acid levels within the normal range were used for the experiment. One hundred of these rats were administered a 0.15g / ml potassium oxonate solution at a dose of 1ml / 100g body weight via gavage to establish the hyperuricemia model, once daily, for continuous replication. Ten rats served as a normal control group, receiving distilled water via gavage under the same conditions. Blood was collected from the orbital sinus of the rats on day 7 of modeling to measure uric acid levels; a two-fold increase in uric acid confirmed a successful model. The successfully modeled rats were then evenly divided into ten groups based on their uric acid levels: a model control group, a positive drug group, and two dosage groups for each of the four processes (process 1, process 2, process 3, and process 4), with ten rats in each group. All treatment groups began gavage administration at 1ml / 100g once daily for 14 consecutive days. The normal control and model control groups received distilled water via gavage under the same conditions.

[0074] Testing parameters: Patients should fast the night before blood collection; blood samples will be collected on the mornings of days 7 and 14 of drug administration to measure uric acid (UA), triglycerides (TG), and total cholesterol (TC). Drug administration will be carried out in the afternoon.

[0075] 3. Experimental Results Table 4. Effects of different processing methods of Geshu granules on a rat model of hyperuricemia.

[0076] Note: Compared with the normal control group ## P <0.01; compared with the model control group, P <0.01.

[0077] Table 4 shows that serum uric acid levels in all groups were significantly elevated after modeling (before drug administration), with a significant difference compared to the normal control group. P <0.01), and there was no difference between the groups. During the experiment, the serum uric acid level in the normal control group remained stable, while the level in the model control group remained elevated, both showing significant differences compared to the normal control group. P <0.01); From day 7 to day 14 after administration of Geshu granules, both dosage groups of the four process samples significantly reduced the serum uric acid level in rats, showing significant differences compared with the model control group. P <0.01), among which process three has the best efficacy.

[0078] Table 5. Effects of different processing methods of Geshu granules on a rat model of hyperuricemia.

[0079] Note: Compared with the normal control group, ##P<0.01; compared with the model control group, P <0.01, P <0.05.

[0080] Table 5 shows that the serum triglyceride levels in the normal control group remained stable during the experiment; the levels in the model control group remained significantly elevated, showing a significant difference compared to the normal control group. P <0.01); After 7 days of administration of the low-dose group of Process 1, 7 to 14 days of administration of the high-dose group of Process 3, and 14 days of administration of the low-dose group of Process 4, the serum triglyceride level in rats was significantly reduced, and the differences were statistically significant compared with the model control group. P <0.01), among which process three has the best efficacy.

[0081] Table 6. Effects of different processing methods of Geshu granules on a rat model of hyperuricemia.

[0082] Note: Compared with the normal control group, ##P<0.01; compared with the model control group, P <0.01.

[0083] Table 6 shows that the serum cholesterol levels in the normal control group remained stable during the experiment; the serum cholesterol levels in the model control group remained at a significantly elevated level, showing a significant difference compared with the normal control group. P <0.01); From day 7 to day 14 after administration of Geshu granules, both dosage groups of the four process samples significantly reduced the blood cholesterol levels in rats, showing significant differences compared with the model control group. P <0.01), among which process three has the best efficacy.

[0084] 4. Conclusion Four different preparation processes of Geshu granules were divided into two dosage groups: one equivalent to the clinical dosage and the other half equivalent. After 14 days of continuous administration, all dosage groups of process 1, process 2, process 3 and process 4 showed a significant reduction in blood uric acid, blood triglyceride and blood cholesterol levels, with process 3 showing the best efficacy.

[0085] Example 2 Optimization of the method for preparing the kudzu root drug composition based on process three 1. Water extraction process 1.1 Orthogonal Experiment of Water Extraction Process When optimizing the water extraction process, the three factors of extraction times, extraction time, and solvent usage were optimized at three levels. The content of berberine in Phellodendron bark, astilbin in Smilax glabra, and the yield of dry extract were used as evaluation indicators to establish a comprehensive scoring standard.

[0086] Weigh out 50g of Phellodendron bark and 75g of Smilax glabra, mix them, and extract by reflux in an electric heating mantle. Conduct the experiment according to the orthogonal design factor table of the water extraction process in Table 1. Use L9(3 4 The experiment was arranged using an orthogonal array. The factor level table and orthogonal experimental design are shown in Table 7, and the specific experimental details are shown in Table 8.

[0087] Table 7. Orthogonal Design Factors for Water Extraction Process

[0088] Table 8 Orthogonal Experiment Table

[0089] 1.2 Content Determination 1.2.1 Berberine Chromatographic conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the stationary phase; acetonitrile-0.1% phosphoric acid solution (50:50) (with 0.1 g sodium dodecyl sulfonate per 100 ml) was used as the mobile phase; the detection wavelength was 265 nm. The theoretical plate number, calculated based on the berberine hydrochloride peak, should be no less than 4000.

[0090] Preparation of reference solution: Take an appropriate amount of berberine hydrochloride reference standard, accurately weigh it, and add it to the mobile phase to prepare a solution containing 0.1 mg per ml.

[0091] Preparation of the test solution: Weigh approximately 0.1 g of the powder (passed through a No. 3 sieve) accurately, place it in a 100 ml volumetric flask, add 80 ml of mobile phase, sonicate (power 250 W, frequency 40 kHz) for 40 min, cool, dilute to the mark with mobile phase, shake well, filter, and collect the filtrate to obtain the test solution.

[0092] Extraction solution: Accurately pipette an appropriate amount of orthogonal aqueous extract, concentrate under reduced pressure below 50℃, or filter directly through a 0.22μm filter membrane to obtain the extract.

[0093] Assay: Accurately pipette 5 μl of the reference solution and 5–20 μl of the test solution into the liquid chromatograph and determine the result.

[0094] 1.2.2 Astilbene Chromatographic conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the stationary phase; methanol-0.1% glacial acetic acid solution (39:61, v / v) was used as the mobile phase; the detection wavelength was 291 nm. The theoretical plate number, calculated based on the astilbin peak, should be no less than 5000.

[0095] Preparation of reference solution: Take an appropriate amount of astilbene reference standard, accurately weigh it, and add 60% methanol to prepare a solution containing 0.2 mg per ml.

[0096] Preparation of the test solution: Take about 0.8g of the powder (passed through a No. 2 sieve), accurately weigh it, place it in a round-bottom flask, accurately add 100ml of 60% methanol, weigh it, heat under reflux for 1 hour, cool it, weigh it again, make up the lost weight with 60% methanol, shake well, filter it, and take the filtrate to obtain the test solution.

[0097] Extraction solution: Accurately pipette an appropriate amount of orthogonal aqueous extract, concentrate under reduced pressure below 50℃, or filter directly through a 0.22μm filter membrane to obtain the extract.

[0098] Assay: Accurately pipette 10 μl each of the reference solution and the test solution into the liquid chromatograph and determine the result.

[0099] 1.3 Determination of dry extract Accurately measure 50 mL of each of the nine samples and place them in a dry, constant-weight evaporating dish. Evaporate to dryness in a water bath and dry in an oven at 105℃ for 5 hours. Remove the samples and cool them in a desiccator for 30 minutes. Remove the samples and weigh them quickly and accurately. Calculate the yield of the dry extract for each of the nine samples.

[0100] 1.4 Analysis of Orthogonal Experiments for Water Lifting Table 9 Results of the orthogonal experiment on water lifting

[0101] The orthogonal results show that A3B3C2 is the optimal process, which is 8 times the amount of water, 3 extractions, 2 hours each time.

[0102] 1.5 Validation of water extraction process Weigh out 100g of Phellodendron bark and 150g of Smilax glabra and mix them together; extract according to Table 10: add 8 times the amount of water for the first extraction, and 6 times the amount of water for the second and third extractions. Measure the volume of the liquid, take 50ml of each, dry them, and calculate the yield of the dry extract; take another 50ml for content detection and retain the sample.

[0103] Table 10 Results of the re-validation of the water extraction process

[0104] Results: The transfer rates were comparable between extractions of 2 and 3 times, with no significant difference. Therefore, the number of extractions was set at 2.

[0105] 1.6 Re-validation of water extraction process Weigh out 100g of Phellodendron bark and 150g of Smilax glabra and mix them together; extract according to Table 11: add 8 times the amount of water for the first extraction and 6 times the amount of water for the second extraction.

[0106] Table 11 Validation results of water extraction process

[0107] As shown in Table 5, the yields of the extracts obtained under this water extraction process were 17.37%, 17.16%, and 17.70%, respectively, indicating that the process is stable and feasible.

[0108] 2. Alcohol Extraction Process 2.1 Orthogonal Experiment of Alcohol Extraction Process When optimizing the alcohol extraction process, the four factors of extraction times, extraction time, solvent volume, and ethanol concentration were optimized at three levels. The contents of puerarin in kudzu root, atractylodes in atractylodes, and β-ecdysterone in achyranthes root, as well as the yield of dry extract, were used as evaluation indicators to establish a comprehensive scoring standard.

[0109] Weigh 450g of Atractylodes lancea according to the prescription ratio and extract the volatile oil (coarse powder) under optimized conditions. Add 8 times the amount of water and extract the volatile oil by steam distillation for 6 hours. Collect the volatile oil and the liquid, and store for later use. Divide the residue into 9 equal portions. Mix each portion with 150g of Pueraria lobata and 30g of Achyranthes bidentata, and extract by reflux in an electric heating mantle. Conduct the experiment according to the orthogonal design factor table of alcohol extraction process in Table 12. Use L9(3 4 The experiment was arranged using an orthogonal array. The factor level table and orthogonal experimental design are shown in Table 13.

[0110] Table 12 Orthogonal Design Factors for Alcohol Extraction Process

[0111] Table 13 Orthogonal Experiment Table for Alcohol Extraction

[0112] 2.2 Content Determination 2.2.1 Puerarin Chromatographic conditions and system suitability test. Octadecylsilane-bonded silica gel was used as the stationary phase; methanol-water (25:75, v / v) was used as the mobile phase; the detection wavelength was 250 nm. The theoretical plate number, calculated based on the puerarin peak, should be no less than 4000.

[0113] Preparation of reference solution: Take an appropriate amount of puerarin reference standard, accurately weigh it, and add 30% ethanol to prepare a solution containing 80 μg per ml.

[0114] Take about 0.1g of the powder of this product (passed through a No. 3 sieve), accurately weigh it, place it in a stoppered conical flask, accurately add 50ml of 30% ethanol, weigh it, heat under reflux for 30min, cool it, weigh it again, make up the weight loss with 30% ethanol, shake well, filter it, and take the filtrate to obtain the product.

[0115] Extraction solution: Accurately pipette an appropriate amount of orthogonal aqueous extract, concentrate under reduced pressure below 50℃, or filter directly through a 0.22μm filter membrane to obtain the extract.

[0116] Determination method: Accurately pipette 10 μl of the reference solution and the test solution into the liquid chromatograph and determine the result.

[0117] 2.2.2 Atractylodesin Chromatographic conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the stationary phase; methanol-water (79:21) was used as the mobile phase; and the detection wavelength was 340 nm. The theoretical plate number, calculated based on the atractylodesin peak, should be no less than 5000.

[0118] Preparation of reference solution: Take an appropriate amount of atractylodes reference standard, accurately weigh it, and add methanol to prepare a solution containing 20 μg per ml.

[0119] Medicinal material: Take about 0.2g of the powder (passed through a No. 3 sieve), accurately weigh it, place it in a stoppered conical flask, accurately add 50ml of methanol, seal tightly, weigh it, sonicate (power 250W, frequency 40kHz) for 1 hour, cool, weigh it again, make up the lost weight with methanol, shake well, filter, and take the filtrate to obtain the product.

[0120] Extraction solution: Accurately pipette an appropriate amount of orthogonal aqueous extract, concentrate under reduced pressure below 50℃, or filter directly through a 0.22μm filter membrane to obtain the extract.

[0121] Determination method: Accurately pipette 10 μl of the reference solution and the test solution into the liquid chromatograph and determine the result.

[0122] 2.2.3 β-ecdysterone Chromatographic conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the stationary phase; acetonitrile-water-formic acid (16:84:0.1) was used as the mobile phase; the detection wavelength was 250 nm. The theoretical plate number, calculated based on the β-ecdysterone peak, should be no less than 4000.

[0123] Preparation of the reference solution: Weigh an appropriate amount of β-ecdysterone reference standard accurately, add methanol to prepare a solution containing 0.1 mg per ml.

[0124] Medicinal Material: Take about 1g of the powder (passed through a No. 3 sieve), accurately weigh it, place it in a stoppered conical flask, add 30ml of water-saturated n-butanol, seal tightly, soak overnight, sonicate (power 300W, frequency 40kHz) for 30min, filter, wash the container and residue several times with 10ml of methanol, combine the filtrate and washings, evaporate to dryness, dissolve the residue in methanol, transfer to a 5ml volumetric flask, add methanol to the mark, shake well, and the product is ready.

[0125] Extraction solution: Accurately pipette an appropriate amount of orthogonal aqueous extract, concentrate under reduced pressure below 50℃, or filter directly through a 0.22μm filter membrane to obtain the extract.

[0126] Determination method: Accurately pipette 10 μl of the reference solution and the test solution into the liquid chromatograph and determine the result.

[0127] 2.3 Determination of dry extract Accurately measure 50 mL of each of the nine samples and place them in a dry, constant-weight evaporating dish. Evaporate to dryness in a water bath and dry in an oven at 105℃ for 5 hours. Remove the samples and cool them in a desiccator for 30 minutes. Remove the samples and weigh them quickly and accurately. Calculate the yield of the dry extract for each of the nine samples.

[0128] Table 14 Results of the orthogonal experiment for alcohol extraction

[0129] Orthogonal result: D3A2C3B3, which is 10 times the amount of 70% ethanol extracted, extracted twice, 2 hours each time.

[0130] 2.4 Validation of the alcohol extraction process Weigh 150g of the residue after extracting the volatile oil from Atractylodes lancea according to the prescription ratio, and divide it into 3 equal portions. Each portion of residue is mixed with 150g of Pueraria lobata and 30g of Achyranthes bidentata, respectively, for alcohol extraction process verification. As shown in Table 15, the extract yields under this extraction process are 14.20%, 15.96%, and 16.24%, respectively, indicating that the alcohol extraction process is stable and feasible.

[0131] Table 15 Validation Results of Alcohol Extraction Process

[0132] 3. Volatile oil extraction process 3.1 Optimization of extraction process (1) Direct extraction of Atractylodes lancea slices Take 100g of Atractylodes lancea and divide it into 4 portions. Place each portion in a round-bottom flask and add 4, 6, 8, and 10 times the amount of water, respectively, to extract the volatile oil. Read the oil volume in the extractor every hour (ml). The experimental results are shown in Table 16.

[0133] Table 16. Investigation of oil extraction rate under different water addition amounts and extraction times

[0134] As shown in Table 10, the optimal extraction process for Atractylodes lancea volatile oil is extraction with 8 times the amount of water for 6 hours.

[0135] (2) Extract Atractylodes lancea powder from raw Atractylodes lancea slices by crushing them into co Take 100g of Atractylodes lancea, grind it into four coarse powder portions, place them in round-bottom flasks, and add 4, 6, 8, and 10 times the amount of water respectively to extract the volatile oil. Read the oil volume in the volatile oil extractor in ml every hour. The experimental results are shown in Table 17.

[0136] Table 17. Investigation of oil extraction rate of Atractylodes lancea coarse powder under different water addition amounts and extraction times.

[0137] The overall results show that the volatile oil was basically completely extracted after 6 hours of extraction, and the volatile oil extraction yield was the highest when 8 times the amount of water was added to the coarse Atractylodes lancea powder.

[0138] As shown in Tables 16 and 17, the yield of volatile oil is higher after pulverizing Atractylodes lancea. Therefore, the optimal extraction process for volatile oil from Atractylodes lancea is: pulverize Atractylodes lancea into coarse powder, add 8 times the amount of water, and extract for 6 hours.

[0139] 3.2 Validation of volatile oil extraction process Take 200g of Atractylodes lancea in 3 portions, pulverize them, add 8 times the amount of water to each portion, and extract the volatile oil for 6 hours. Dry the liquid at 70℃.

[0140] Table 18 Validation Results of Volatile Oil Extraction Process

[0141] As shown in Table 18, under the same feeding conditions, the amount of volatile oil obtained by the volatile oil extraction process is basically the same. Therefore, the volatile oil extraction process of Atractylodes lancea is stable and feasible.

[0142] 3.3 Investigation on the Encapsulation Process of Atractylodes lancea Volatile Oil 3.3.1 Optimal selection of encapsulation process (1) Water-saturated solution method Weigh 32g of β-cyclodextrin (β-CD), add 320ml of water, and dissolve until clear at 60℃ (or prepare a saturated aqueous solution). Cool to 40℃, and slowly add 4.0g of volatile oil while stirring. Stir magnetically at 40℃ for 2 hours (750 rpm). -1 The precipitate was placed at room temperature, refrigerated overnight, filtered, and dried in an oven at 40°C for 4 hours to obtain the inclusion complex.

[0143] (2) Grinding method Weigh 24.0g of β-CD, add 4.0g of volatile oil, grind, mix well, air dry (dry at low temperature 40℃), grind into powder, and weigh.

[0144] (3) Ultrasonic method Weigh 24.0g of β-CD, add it to 144mL of water, heat in a water bath to dissolve, and when the temperature drops to 40℃, add 4g of volatile oil, shake thoroughly, sonicate for 30min, refrigerate for 24h, filter, collect the precipitate, and dry at 40℃ to obtain the inclusion complex.

[0145] (4) Colloid milling method Weigh 57.6g of β-CD, add 172.8mL of water, heat and mix until uniform at 30℃ to form a suspension, then add 7.2g of volatile oil, and slowly add it to a colloid mill while stirring. Grind for 30min, release the material, refrigerate overnight, centrifuge, take the inclusion complex, and dry in a 40℃ oven for 4h.

[0146] Note: The above cyclodextrin aqueous solution should not be stirred during the cooling process; it should be allowed to cool naturally.

[0147] Table 19 Examination of Encapsulation Process

[0148] 3.3.2 Stability Study of Volatile Oil Inclusion Compounds Take the above inclusion complex, put it into an aluminum plastic sealed bag (simulating market packaging), each bag contains 3.0g, and place it in a refrigerator, room temperature (23~30℃), (40±5)℃, and (60±5)℃ environments for 10 days respectively. Determine the content of β-cineole, atractylone, and atractylodes in the inclusion complex. Take about 1g of each sample, weigh it accurately, put it into a pre-weighed weighing bottle, and place it in an open oven at (40±5)℃ and (60±5)℃ for 10 days. Weigh it accurately again and determine the content of each index component. The results are shown in Table 20.

[0149] Table 20 Total content of β-cineole, atractylone, and atractylone in inclusion complex after 10 days of storage under different conditions.

[0150] Tables 19 and 20 show that the inclusion rates of water-saturated, grinding, and ultrasonic methods are all higher than those of colloid milling. After 10 days of storage under different conditions, the main index components of the four inclusion methods all decreased, with the water-saturated solution method and ultrasonic method showing the slowest decrease in index components and higher retention rates. Therefore, from a practical production perspective, the water-saturated solution method is preferred. 3.3.3 Orthogonal Experiment of Water-Saturated Solution Using the inclusion rate of volatile oil and the yield of inclusion products as indicators, according to L9(3 4 An orthogonal design table was used for the experiment, with inclusion temperature, volatile oil (g): β-CD (g): water and inclusion time as the factors to be investigated. Each factor was tested at three different levels, as shown in Table 21.

[0151] Table 21 Level of Factors in Volatile Oil Encapsulation Process

[0152] Inclusion complex recovery rate determination: Inclusion complexes were obtained from each sample, placed in a round-bottom flask, 300 ml of water was added, the volatile oil extractor was connected, and the mixture was slowly heated to boiling using a temperature-controlled electric heating mantle. After extraction for 5 hours, the mixture was allowed to stand for 30 minutes, and the volume of volatile oil was recorded.

[0153] Volatile oil yield = (Volatile oil recovered amount / Volatile oil added amount) × Blank recovery rate × 100%; Inclusion complex yield = (Total inclusion complex / (Amount of volatile oil added + Amount of cyclodextrin added)) × 100%; Overall score = (volatile oil yield / highest yield × 0.7 + inclusion compound yield / highest yield × 0.3) × 100%.

[0154] Table 21 shows that, based on intuitive analysis and comprehensive scoring, the optimal process is an inclusion temperature of 50℃, a volatile oil volume to β-cyclodextrin volume ratio of 1:12, and an inclusion time of 3 hours. Orthogonal analysis results indicate that B3A3C3, i.e., volatile oil (g):cyclodextrin (g):water (g) = 1:12:120, inclusion temperature of 50℃, and inclusion time of 3 hours, represents the optimal process.

[0155] Based on the above analysis, the optimal process for inclusion of volatile oil is as follows: volatile oil (g): cyclodextrin (g): water (g) = 1:12:120, inclusion temperature 50℃, and inclusion time 3h.

[0156] Table 22 Levels of Process Factors for Volatile Oil Encapsulation

[0157] 4. Concentration and drying temperature investigation Sample concentration was performed at the factory's standard concentration temperature of 70℃, with a concentration density of 1.30~1.35. Drying temperatures were 60℃, 70℃, 80℃, and 90℃, respectively. Based on the content of the main indicator components, it is recommended that the sample drying temperature not exceed 80℃.

[0158] 5 Formulation Process 5.1 The evaluation of fillers is shown in Table 23.

[0159] Table 23 Results of preparing granules with different filler combinations

[0160] Currently, commonly used fillers in granules are sucrose, lactose, and dextrin. Considering that patients with hyperuricemia may also have diabetes, this product is intended to be sugar-free, excluding sucrose as a filler. Through optimization of different proportions of dextrin and lactose, it was found that without fillers, the extract of this composition exhibited good formability, with a formability rate of 95.43%. Dextrin, as a filler, showed better formability than lactose and was relatively cheaper; therefore, in actual production, if a filler is required, dextrin is the preferred choice.

[0161] As shown in Table 23, the molding rate is good when the amount of dextrin added is 20%, 30% and 40% of the total formulation. Therefore, when dextrin is used as a filler, the molding properties of dextrin are good, and the proportion of dextrin can be added according to actual needs.

[0162] 5.2 Exploration of Flavoring in Pharmaceutical Formulations The drug composition of Geshu contains Phellodendron bark and Atractylodes lancea. Phellodendron bark is bitter and Atractylodes lancea is pungent, and it also contains volatile oil inclusion compounds. In order to mask the bitterness and spiciness, this experiment used stevioside, mannitol and citric acid to correct the taste and aroma by searching, collecting, selecting and analyzing flavoring agents and masking agents, and compared the taste.

[0163] By comparing the effects of adding different proportions of stevia, mannitol, and citric acid, mixing the mixtures, dissolving 5g of each mixture in 200ml of water (50℃), and tasting the samples, the results are shown in Table 24. Ultimately, adding 1% stevia and 0.5% anhydrous citric acid significantly improved the bitterness and spiciness, resulting in a better taste that is generally acceptable.

[0164] Table 24 Flavoring results of different sweeteners

[0165] Example 3 Effects of the kudzu root drug combination on a rat model of hyperuricemia combined with acute gouty arthritis induced by adenine and yeast 1. Experimental Materials 1.1 Test Drug Geshu granules (dry extract powder), sample prepared by process three; batch number: 2502001; provided by the New Drug R&D Center of the Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences. Appearance: brownish-yellow powder; production date: February 28, 2025; shelf life: 2 years. Sample drug content: 3.6g crude drug / g extract powder; storage conditions: room temperature, sealed.

[0166] 1.2 Positive control drugs are shown in Table 25. Table 25

[0167] 1.3 The experimental animals are shown in Table 26.

[0168] Table 26

[0169] 1.4 The test instruments are shown in Table 27.

[0170] Table 27

[0171] 1.5 Test reagents are shown in Table 28.

[0172] Table 28

[0173] 2. Drug Dosage Design 2.1 Test Drug In this study, rats were administered 6.39 g crude drug / kg, 3.20 g crude drug / kg / day, and 1.60 g crude drug / kg / day, which are equivalent to multiples, 1 / 2 times, and 1 / 4 times the human clinical dosage, respectively. All dosages of Ge Shu granules in the experiment were calculated based on the amount of crude drug.

[0174] Geshu granules were administered as a dry extract powder, with each 1g of dry extract powder containing 3.6g of raw herb. Based on the amount of raw herb in each dosage group, 35.6g, 17.8g, and 8.9g of dry extract powder were taken from the high, medium, and low dosage groups, respectively, and each was diluted with distilled water to 200mL to prepare dry extract powder suspensions with concentrations of 0.18g / mL, 0.9g / mL, and 0.45g / mL, respectively, for administration. During the experiment, rats were administered the drug by gavage at a dose of 1ml / 100g once daily for 4 consecutive weeks.

[0175] 2.2 Positive control drug The adult clinical dose of benzbromarone tablets, a positive control drug, is 50 mg / day. Based on body surface area, the rat dose is 4.5 mg / kg / day, which is equivalent to a multiple of the human clinical dose.

[0176] In the benzbromarone group, 100 mg of benzbromarone tablets (50 mg / tablet) were ground and diluted with distilled water to a final volume of 222 mL to prepare a suspension with a concentration of 0.45 mg / mL for administration. During the experiment, rats were administered the drug by gavage at a dose of 1 ml / 100 g once daily for 4 consecutive weeks.

[0177] The positive control drug for traditional Chinese medicine, Tongfengshu tablets, is used clinically for adults at a dose of 2-4 tablets (0.32g / tablet) three times a day. The experimental dosage was calculated based on 3 tablets three times a day. The dosage for rats was 0.26 g / kg / day, which is equivalent to a multiple of the human clinical dose, based on the body surface area.

[0178] In the gout relief group, 4.8g of gout relief tablets (15 tablets) were ground and diluted with distilled water to a final volume of 185mL to prepare a suspension with a concentration of 0.026 g / mL for administration. During the experiment, rats were administered the drug by gavage at a dose of 1ml / 100g once daily for 4 consecutive weeks.

[0179] 2.3 Drugs for creating a model of hyperuricemia Adenine was prepared at a concentration of 10 mg / mL before the test. During the test, it was administered by gavage at a dose of 1 ml / 100 g body weight once a day for 7 consecutive weeks.

[0180] 2.4 Drugs for creating models of acute arthritis Sodium urate was administered via intra-articular injection of 0.1 ml per rat in the right ankle of a 100 mg / mL sodium urate suspension.

[0181] 3. Experimental methods and results 3.1 Modeling method and drug administration Eighty male SD rats (SPF grade, weighing 240-260g) were used. Baseline uric acid levels were measured by orbital blood sampling before the experiment. Seventy rats with uric acid levels within the normal range were selected for the experiment. Sixty of these rats were administered a 10mg / ml adenine solution via gavage at a rate of 1ml / 100g body weight, along with a diet containing 10% yeast, at a rate of 100g / kg / day (each group was fed the full amount separately, supplemented with regular feed after complete consumption). This process was repeated for 7 weeks to establish a hyperuricemia model. The remaining 10 rats served as a normal control group, receiving distilled water via gavage and a regular diet under the same conditions. Blood uric acid (UA) levels were measured by orbital blood sampling at week 3 of modeling. A UA level that increased by more than 1 times was considered a successful model.

[0182] Animals that successfully developed the uric acid model were randomly divided into four groups based on their serum uric acid levels: a model control group, a benzbromarone group, a gout relief group, and high-dose, medium-dose, and low-dose groups of Geshu granules, with 10 animals in each group. Each treatment group was then administered the drug via gavage at a dose of 1 ml / 100g body weight, once daily for four consecutive weeks. The normal control group and the model control group were administered distilled water via gavage under the same conditions. The animals were dissected at week 4, and the following indicators were observed: ① Two weeks after drug administration, animals were anesthetized, and blood was collected from the orbital cavity to measure the levels of uric acid (UA), triglycerides (TG), cholesterol (CHO), blood urea nitrogen (UREA), creatinine (CRE), blood glucose (GLU), C-reactive protein (CRP), and homocysteine ​​(Hcy).

[0183] ② Four days before dissection in the fourth week after drug administration, the volume of the right posterior ankle joint was measured using a toe edema meter. Then, except for the normal control group, all other groups of animals were injected with 100 μl / animal of a 100 mg / ml sodium urate solution into the right ankle joint cavity to establish an acute gouty arthritis model. The normal control group was injected with physiological saline under the same conditions. The volume of the right ankle joint was measured on the third day after modeling.

[0184] ③ In the fourth week after administration, the animals were anesthetized, blood was collected from the abdominal aorta, and the levels of uric acid (UA), triglycerides (TG), cholesterol (CHO), blood urea nitrogen (UREA), creatinine (CRE), blood glucose (GLU), C-reactive protein (CRP), homocysteine ​​(Hcy), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) were measured.

[0185] ④ Dissect the animal and take tissue from the right posterior ankle joint, liver, and kidney for pathological and histological examination.

[0186] 3.2 Data Statistics Measurement data -x±s indicates that a one-way ANOVA was used, and pairwise comparisons were performed using the t-test. P <0.05, P <0.01 indicates that the difference is statistically significant.

[0187] 3.3 Test Results Table 29 Effects on a rat model of hyperuricemia complicated with acute gouty arthritis induced by adenine and yeast

[0188] Note: Compared with the normal control group: ##P<0.01; Compared with the model control group: P<0.01.

[0189] Table 29 shows that: before modeling, the serum uric acid levels in each group were uniform and without difference; after modeling (before drug administration), the serum uric acid levels in each group of rats increased significantly, showing significant differences compared with the normal control group (P<0.01), and there were no differences among the groups; after 2 and 4 weeks of administration of the positive control drugs benzbromarone, Gout Relief, and Geshu Granules, the serum uric acid levels in all three dosage groups decreased significantly, showing significant differences compared with the model control group (P<0.01). P <0.01).

[0190] Table 30 Effects on a rat model of hyperuricemia complicated with acute gouty arthritis induced by adenine and yeast

[0191] Note: Compared with the normal control group: ## P <0.01; compared with the model control group: P <0.05, P <0.01.

[0192] Table 30 shows that the serum levels of CRP and Hcy in the model control group rats were significantly elevated compared with those in the normal control group. P <0.01); After 2 and 4 weeks of administration of the three dose groups of the positive control drugs benzbromarone, Gout Relief, and Geshu Granules, serum CRP and Hcy levels were significantly reduced, and the differences were statistically significant compared with the model control group. P <0.05, P <0.01).

[0193] Table 31 Effects on a rat model of hyperuricemia complicated with acute gouty arthritis induced by adenine + yeast (drug administration for 2 weeks)

[0194] Note: Compared with the normal control group ## P <0.01; compared with the model control group: P <0.01.

[0195] Table 32 Effects on a rat model of hyperuricemia complicated with acute gouty arthritis induced by adenine + yeast (drug administration for 4 weeks)

[0196] Note: Compared with the normal control group ## P <0.01; compared with the model control group: P <0.01.

[0197] Tables 31 and 32 show that serum TG, TC, CRE, UREA, and GLU levels in the model control group were significantly elevated compared to the normal control group. P <0.01); the positive control group, benzbromarone, Gout Relief, and Geshu Granules at all three dosage groups significantly reduced the levels of TG, TC, CRE, UREA, and GLU in rat serum, showing significant differences compared with the model control group. P <0.01).

[0198] Table 33 Effects on a rat model of hyperuricemia complicated with acute gouty arthritis induced by adenine + yeast (drug administration for 4 weeks)

[0199] Note: Compared with the normal control group ## P <0.01; compared with the model control group: P <0.01.

[0200] Table 33 shows that serum AST and ALT levels in the model control group were significantly elevated compared to the normal control group. P <0.01); After four weeks of administration of the three dose groups of the positive control drug benzbromarone, Gout Relief, and Geshu Granules, the levels of AST and ALT in the serum of rats were significantly reduced, and the differences were statistically significant compared with the model control group (P<0.01).

[0201] Table 34 Effects on a rat model of hyperuricemia complicated with acute gouty arthritis induced by adenine and yeast

[0202] Note: Compared with the normal control group: ## P<0.01; compared with the model control group: P <0.01.

[0203] Table 34 shows that there was no difference in the volume of the right ankle joint among the groups before modeling. Three days after modeling, the volume of the right ankle joint of the rats in the model control group was significantly increased, which was significantly different from that of the normal control group (P<0.01). The three dose groups of the positive control drugs benzbromarone, Tongfengshu, and Geshu granules could significantly reduce the volume of the right ankle joint of the rats after modeling, which was significantly different from that of the model control group (P<0.01).

[0204] 4. Conclusion Geshu granules have a good therapeutic effect on a rat model of adenine + yeast-induced hyperuricemia complicated with acute gouty arthritis. They can reduce the levels of UA, TG, TC, CRE, UREA, GLU, ALT and AST in the blood; reduce CRP and Hcy levels; and reduce ankle swelling.

[0205] Example 4 Rat liver and kidney pathology report Experimental Objective The pathological effects of Geshu granules on the kidneys and liver were observed and evaluated in rats with hyperuricemia and acute gouty arthritis.

[0206] Test methods Animals: Rats in each group of Example 3.

[0207] HE staining was performed, and the pathological changes in rat kidney and liver tissues were observed under a light microscope and photographed. Testing unit: Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences.

[0208] The histopathological grading of rat kidney tissue was based on the following criteria: ① renal tubular injury; ② glomerular injury; ③ interstitial fibrosis; ④ the extent and degree of interstitial inflammatory cell infiltration. The lesions were classified into five grades. Table 35. Grading Criteria for Kidney Histopathology

[0209] The histopathological grading of rat liver tissue was based on the extent and degree of hepatocyte vacuolation, classifying the lesions into 5 grades: Table 36. Grading Criteria for Liver Histopathology

[0210] Statistical analysis: SPSS 20.0 statistical software was used for statistical analysis. Quantitative data were expressed as mean ± standard deviation (x ± s). One-way ANOVA and nonparametric rank-sum tests were used for comparisons between groups.P <0.05 is the standard for significance testing.

[0211] Test results are shown Figure 5 and Figure 6 .

[0212] The results of the pathological light microscopy examination of the kidneys are as follows: 1. Normal control group: The rat kidneys were intact, the basement membrane was uniform, no mesangial cell proliferation was observed, the renal tubular epithelial cells were tightly arranged, the brush border was clear, there were no casts or crystal deposits, and there was no fibrous tissue proliferation or inflammatory cell infiltration in the interstitium.

[0213] 2. Model control group: Rats showed mild to severe proliferation of the glomerular mesangial matrix, partial glomerular sclerosis, degeneration / necrosis and atrophy of proximal tubular epithelial cells, thickening of the basement membrane, and protein casts and urate crystals in some tubular lumens. The renal interstitium showed mild to moderate fibrosis and multifocal inflammatory cell infiltration, mainly mononuclear cells, which was significantly different from the normal control group. P <0.01).

[0214] Compared with the model control group, kidney damage was reduced to varying degrees in all drug-treated groups, including: 3. Western medicine control group, and high, medium, and low dose groups of Geshu granules: The degree of mesangial matrix deposition in rat glomeruli was significantly reduced, the extent of renal tubular degeneration / necrosis was significantly reduced, and the extent and degree of interstitial fibrosis and inflammatory cell infiltration were significantly reduced. P <0.05 or <0.01) indicates a significant protective effect against kidney damage.

[0215] 4. Traditional Chinese medicine control group: The extent and degree of glomerular damage, renal interstitial fibrosis, and inflammatory cell infiltration in rats were significantly reduced. P <0.05 or P <0.01), mild to moderate degeneration / necrosis of renal tubules, with a decreasing trend in the score ( P <0.1), indicating that it also has a significant protective effect against kidney damage.

[0216] The degree of protection against kidney injury in mice, from strongest to weakest, is as follows: medium-dose Geshu granules group > low-dose Geshu granules group > Western medicine control group > high-dose Geshu granules group > traditional Chinese medicine control group.

[0217] The results of the pathological light microscopy examination of the liver are as follows: 1. Normal control group: The liver lobule structure of rats was clear, the hepatocytes were of uniform size and arranged radially with the central vein as the center. No degeneration or necrosis was observed, and no fibrous tissue hyperplasia or inflammatory cell infiltration was observed in the interstitium. The tissue structure was normal.

[0218] 2. Model control group: Rat hepatocytes showed swelling and degeneration, with mild to moderate vacuolation of the cytoplasm, which was significantly different from the normal control group. P <0.01).

[0219] 3. In the Western medicine control group and the high, medium, and low dose groups of Geshu granules: the extent and degree of vacuolation of rat hepatocytes were significantly reduced compared with the model control group. P <0.05 or P <0.01), indicating a significant protective effect against liver damage.

[0220] 4. Traditional Chinese medicine control group: The score of rat hepatocyte vacuolation was lower than that of the model control group. P＞ (0.05), suggesting a slight protective effect against liver damage.

[0221] The protective effect of each group against liver injury in rats, from strongest to weakest, is as follows: high-dose Geshu granules group > medium-dose Geshu granules group > Western medicine control group > low-dose Geshu granules group > traditional Chinese medicine control group.

[0222] Table 37 Statistical analysis of pathological changes in rat renal tubular injury observed under a microscope

[0223] Note: Compared with the model group, P <0.05, P <0.01, # P <0.1.

[0224] Table 38 Statistical analysis of pathological changes in rat glomerular injury observed under a microscope

[0225] Note: Compared with the model group, P <0.05, P <0.01, # P <0.1.

[0226] Table 39 Statistical analysis of pathological changes in rat renal interstitial fibrosis tissue under microscopic observation

[0227] Note: Compared with the model group, P <0.05, P <0.01, # P <0.1.

[0228] Table 40 Statistical analysis of pathological changes in inflammatory cell infiltration in rat renal interstitium observed under a microscope.

[0229] Note: Compared with the model group, P <0.05, P <0.01.

[0230] Table 41 Statistical analysis of pathological changes in rat hepatocyte vacuolation observed under a microscope

[0231] Note: Compared with the model group, P <0.05, P <0.01.

[0232] in conclusion: 1. Normal control group: The rat kidneys were intact, with a uniform basement membrane, no mesangial cell proliferation, tightly packed renal tubular epithelial cells, clear brush borders, no casts or crystal deposits, and no interstitial fibrosis or inflammatory cell infiltration; the liver lobules were clearly structured, the hepatocytes were uniform in size, arranged radially around the central vein, and no degeneration or necrosis was observed, with no interstitial fibrosis or inflammatory cell infiltration.

[0233] 2. Model control group: Rats showed mild to severe proliferation of the glomerular mesangial matrix, partial glomerular sclerosis, degeneration / necrosis and atrophy of proximal tubular epithelial cells, thickening of the basement membrane, and protein casts and urate crystals in some tubular lumens. Mild to moderate fibrosis and multifocal inflammatory cell infiltration, mainly mononuclear cells, were observed in the renal interstitium. Hepatocytes were swollen and degenerated, with mild to moderate vacuolation of the cytoplasm.

[0234] 3. Compared with the model control group, the Western medicine control group and the high, medium, and low dose groups of Geshu granules showed: the degree of mesangial matrix deposition in rat glomeruli was significantly reduced, the extent of renal tubular degeneration / necrosis was significantly reduced, and the degree of interstitial fibrosis and inflammatory cell infiltration was significantly reduced. The extent and degree of hepatocyte vacuolation were also significantly reduced. This suggests that both the Western medicine control group and the various Geshu granule treatment groups have a significant protective effect against liver and kidney tissue damage.

[0235] 4. Traditional Chinese Medicine Control Group: In rats, the extent and degree of glomerular injury, renal interstitial fibrosis, and inflammatory cell infiltration were significantly reduced, while renal tubules showed mild to moderate degeneration / necrosis, with a decreasing score trend, indicating a significant protective effect against kidney injury. The score for hepatocyte vacuolation was reduced, suggesting a slight protective effect against liver injury.

[0236] Normal control group: The rat kidneys were structurally intact, with a uniform basement membrane, no mesangial cell proliferation, tightly packed renal tubular epithelial cells, a clear brush border, no casts or crystal deposits, and no interstitial fibrosis or inflammatory cell infiltration. The liver lobules were clearly structured, with hepatocytes of uniform size, arranged radially around the central vein, without degeneration or necrosis, and no interstitial fibrosis or inflammatory cell infiltration.

[0237] Example 5 Rat joint pathology report A total of 70 samples of HE-stained ankle joint tissue from each group of rats in Example 3 were submitted for testing. Detailed information is shown in Table 42. The testing institution was the Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences.

[0238] Table 42 Slice Grouping

[0239] Experimental Objective The lesions in the ankle joint tissue were observed to evaluate the therapeutic effect of Geshu granules on a rat model of hyperuricemia complicated with acute gouty arthritis.

[0240] Test methods HE staining was performed, and the pathological changes of rat ankle joint tissue were observed under a light microscope and photographed.

[0241] The histopathological grading of rat ankle joint tissue was based on the following criteria: ① synovial cell proliferation; ② interstitial inflammatory cell infiltration; ③ angiogenesis; and ④ the extent and degree of bone destruction. The lesions were classified into five grades. Table 43. Histopathological Grading Criteria for Ankle Joints

[0242] Statistical analysis: SPSS 20.0 statistical software was used for statistical analysis. When comparing sample means, for continuous data that met the requirements of normality and homogeneity of variance, the t-test of two independent samples or one-way ANOVA was used; for those that did not meet these requirements, the nonparametric rank-sum test was used for statistical analysis of tiered data. P <0.05 is the standard for significance testing.

[0243] The pathological light microscopy results are shown in Figure 7 The details are as follows: 1. Normal control group: The articular chondrocytes of rats were neatly arranged, the cartilage and bone structure were normal, no congestion, edema and abnormal proliferation were observed in the synovium, no thickening of the joint capsule, and no fibrous tissue proliferation and inflammatory cell infiltration in the surrounding soft tissue.

[0244] 2. Model control group: Microscopically, rat synovial cells showed mild to severe proliferation, interstitial congestion and edema, accompanied by infiltration of neutrophils, lymphocytes, and monocytes, fibrinoid exudation, joint cavity narrowing, and significant inflammatory tissue necrosis; small blood vessels dilated and proliferated, pannus formed, and bone tissue was destroyed, resembling insect bites; mild to severe inflammatory cell infiltration and fibrous tissue proliferation were observed around the joints. Significant differences were observed compared to the normal control group. P <0.01).

[0245] 3. Traditional Chinese medicine control group: The extent and degree of synovial cell proliferation, interstitial inflammatory cell infiltration, angiogenesis, and bone destruction in rat joints were significantly reduced compared with the model control group. P <0.05), which has a significant protective effect against gouty arthritis.

[0246] 4. High-dose Geshu granules group: Most animals showed mild or slight joint vascular proliferation and mild bone destruction, which were significantly reduced compared with the model control group. P <0.05), the degree of synovial cell proliferation and inflammatory cell infiltration showed a decreasing trend ( P <0.1), which has a significant protective effect against gouty arthritis.

[0247] 5. Western medicine control group: Mild to moderate proliferation of synovial cells in rat joints, significantly reduced compared to the model control group ( P <0.05), the degree of interstitial inflammatory cell infiltration, angiogenesis and bone destruction was reduced, but the difference was not significant. P >0.05), which has a significant protective effect against gouty arthritis.

[0248] 6. Medium-dose group of Geshu granules: The degree of joint vascular proliferation in rats showed a decreasing trend compared with the model control group. P <0.1), indicating a reduction in synovial cell proliferation, angiogenesis, and bone destruction. P >0.05), which has a slight protective effect against gouty arthritis.

[0249] 7. Low-dose Geshu granules group: The extent and severity of joint tissue lesions in rats were not significantly different from those in the model control group. P >0.05), with no significant effect on gouty arthritis.

[0250] The protective effect of each group against gouty arthritis in rats, from strongest to weakest, is as follows: Traditional Chinese medicine control group > high-dose Geshu granules group > Western medicine control group > medium-dose Geshu granules group.

[0251] Table 44. Statistical analysis of pathological changes in synovial cell proliferation in rat ankle joints under microscopic observation.

[0252] Note: Compared with the model group, P <0.05, P < 0.01, # P <0.1.

[0253] Table 45 Statistical analysis of pathological changes in interstitial inflammatory cell infiltration in rat ankle joints under microscopic observation

[0254] Note: Compared with the model group, P <0.05, P <0.01, # P <0.1.

[0255] Table 46 Statistical analysis of pathological changes in angiogenesis in rat ankle joints under microscopic observation

[0256] Note: Compared with the model group, P <0.05, P <0.01, # P <0.1 Table 47 Statistical analysis of pathological changes in rat ankle joint bone destruction observed under a microscope

[0257] Note: Compared with the model group, P <0.05, P <0.01.

[0258] in conclusion 1. Normal control group: The articular chondrocytes of rats were neatly arranged, the cartilage and bone structure were normal, no congestion, edema and abnormal proliferation were observed in the synovium, no thickening of the joint capsule, and no fibrous tissue proliferation and inflammatory cell infiltration in the surrounding soft tissue.

[0259] 2. Model control group: The rat joint synovial cells showed mild to severe proliferation, interstitial congestion and edema, accompanied by infiltration of neutrophils, lymphocytes and monocytes, fibrinoid exudation, joint cavity narrowing, and a large amount of inflammatory tissue necrosis; small blood vessels dilated and proliferated, pannus formed, and destroyed bone tissue, resembling insect bites; mild to severe inflammatory cell infiltration and fibrous tissue proliferation were observed around the joints, indicating that the rat model of hyperuricemia combined with acute gouty arthritis was successfully established.

[0260] 3. Traditional Chinese medicine control group: The extent and degree of synovial cell proliferation, interstitial inflammatory cell infiltration, angiogenesis and bone destruction in rat joints were significantly reduced compared with the model control group, showing a significant protective effect against gouty arthritis.

[0261] 4. High-dose Geshu granules group: Most animals showed mild or slight proliferation of joint blood vessels and mild bone destruction, which were significantly reduced compared with the model control group. The degree of synovial cell proliferation and inflammatory cell infiltration showed a decreasing trend, and it had a significant protective effect against gouty arthritis.

[0262] 5. Western medicine control group: The synovial cells of rat joints showed mild to moderate proliferation, which was significantly reduced compared with the model control group. The infiltration of interstitial inflammatory cells, angiogenesis and bone destruction were reduced, but the difference was not significant. It has a significant protective effect against gouty arthritis.

[0263] 6. Medium-dose group of Geshu granules: Compared with the model control group, the degree of joint vascular proliferation in rats showed a decreasing trend, and the degree of synovial cell proliferation, vascular proliferation and bone destruction were reduced, which had a slight protective effect on gouty arthritis.

[0264] 7. Low-dose Geshu granules group: The extent and degree of joint tissue lesions in rats were not significantly different from those in the model control group, and had no significant effect on gouty arthritis.

[0265] The protective effect of each group against gouty arthritis in rats, from strongest to weakest, is as follows: Traditional Chinese medicine control group > high-dose Geshu granules group > Western medicine control group > medium-dose Geshu granules group.

[0266] Example 6 Effects of Geshu Granules on a Rat Model of Hyperuricemia Combined with Acute Gouty Arthritis Induced by Potassium Oxygenate and Yeast 1. Experimental Materials 1.1 Test Drug Geshu granules (dry extract powder), a sample prepared by process three; provided by the New Drug R&D Center of the Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences. Batch number: 2502001; Appearance: brownish-yellow powder; Production date: February 28, 2025; Shelf life: 2 years. Sample drug content: 3.6g crude drug / g extract powder; Storage conditions: room temperature, sealed.

[0267] 1.2 The sources of the positive control drug, experimental rats, experimental instruments, and reagents are the same as those described in Example 3.

[0268] 2. Drug Dosage Design 2.1 Test Drug In this embodiment, rats were given 6.39 g crude drug / kg, 3.20 g crude drug / kg / day, and 1.60 g crude drug / kg / day, which are equivalent to equal, 1 / 2, and 1 / 4 times the human clinical dosage, respectively. All dosages of Geshu granules administered in the experiment were based on crude drug amounts.

[0269] Geshu granules were administered as a dry extract powder, with each 1g of dry extract powder containing 3.6g of raw herb. Based on the amount of raw herb in each dosage group, 35.6g, 17.8g, and 8.9g of dry extract powder were taken from the high, medium, and low dosage groups, respectively, and each was diluted with distilled water to 200mL to prepare dry extract powder suspensions with concentrations of 1.8g / mL, 0.9g / mL, and 0.45g / mL, respectively, for administration. During the experiment, rats were administered the drug by gavage at a dose of 1ml / 100g once daily for 4 consecutive weeks.

[0270] 2.2 Positive control drug The adult clinical dose of benzbromarone tablets, a positive control drug, is 50 mg / day. Based on body surface area, the rat dose is 4.5 mg / kg / day, which is equivalent to a multiple of the human clinical dose.

[0271] In the benzbromarone group, 100 mg of benzbromarone tablets (50 mg / tablet) were ground and diluted with distilled water to a final volume of 222 mL to prepare a suspension with a concentration of 0.45 mg / mL for administration. During the experiment, rats were administered the drug by gavage at a dose of 1 ml / 100 g once daily for 4 consecutive weeks.

[0272] The positive control drug for traditional Chinese medicine, Tongfengshu tablets, is used clinically for adults at a dose of 2-4 tablets (0.32g / tablet) three times a day. The experimental dosage was calculated based on 3 tablets three times a day. The dosage for rats was 0.26 g / kg / day, which is equivalent to a multiple of the human clinical dose, based on the body surface area.

[0273] In the gout relief group, 4.8g of gout relief tablets (15 tablets) were ground and diluted with distilled water to a final volume of 185mL to prepare a suspension with a concentration of 0.026 g / mL for administration. During the experiment, rats were administered the drug by gavage at a dose of 1ml / 100g once daily for 4 consecutive weeks.

[0274] 2.3 Drugs for creating a model of hyperuricemia Potassium oxonate: Prepare a potassium oxonate solution with a concentration of 0.15 g / mL before the test. During the test, administer 1 ml / 100 g body weight by gavage once a day for 7 consecutive weeks.

[0275] 2.4 Drugs for creating models of acute arthritis Sodium urate was administered via intra-articular injection of 0.1 ml per rat in the right ankle of a 100 mg / mL sodium urate suspension.

[0276] 3. Experimental methods and results 3.1 Modeling method and drug administration Eighty male SD rats (SPF grade, weighing 240-260g) were used. Baseline uric acid levels were measured by orbital blood sampling before the experiment. Seventy rats with uric acid levels within the normal range were selected for the experiment. Sixty of these rats were administered a 10mg / ml adenine solution via gavage at a rate of 1ml / 100g body weight, along with a diet containing 10% yeast, at a rate of 100g / kg / day (each group was fed the full amount separately, supplemented with regular feed after complete consumption). This process was repeated for 7 weeks to establish a hyperuricemia model. The remaining 10 rats served as a normal control group, receiving distilled water via gavage and a regular diet under the same conditions. Blood uric acid (UA) levels were measured by orbital blood sampling at week 3 of modeling. A UA level that increased by more than 1 times was considered a successful model.

[0277] Animals that successfully developed the uric acid model were randomly divided into four groups based on their serum uric acid levels: a model control group, a benzbromarone group, a gout relief group, and high-dose, medium-dose, and low-dose groups of Geshu granules, with 10 animals in each group. Each treatment group was then administered the drug via gavage at a dose of 1 ml / 100g body weight, once daily for four consecutive weeks. The normal control group and the model control group were administered distilled water via gavage under the same conditions. The animals were dissected at week 4, and the following indicators were observed: ① Two weeks after drug administration, animals were anesthetized, and blood was collected from the orbital cavity to measure the levels of uric acid (UA), triglycerides (TG), cholesterol (CHO), blood urea nitrogen (UREA), creatinine (CRE), blood glucose (GLU), C-reactive protein (CRP), and homocysteine ​​(Hcy).

[0278] ② Four days before dissection in the fourth week after drug administration, the volume of the right posterior ankle joint was measured using a toe edema meter. Then, except for the normal control group, all other groups of animals were injected with 100 μl / animal of a 100 mg / ml sodium urate solution into the right ankle joint cavity to establish an acute gouty arthritis model. The normal control group was injected with physiological saline under the same conditions. The volume of the right ankle joint was measured on the third day after modeling.

[0279] ③ In the fourth week after administration, the animals were anesthetized, and blood was collected from the abdominal aorta to measure the levels of uric acid (UA), triglycerides (TG), cholesterol (CHO), blood urea nitrogen (UREA), creatinine (CRE), blood glucose (GLU), C-reactive protein (CRP), homocysteine ​​(Hcy), alanine aminotransferase (ALT), and aspartate aminotransferase (AST).

[0280] ④ Dissect the animal and take tissue from the right posterior ankle joint, liver, and kidney for pathological and histological examination.

[0281] 3.2 Data Statistics Measurement data -x±s indicates that a one-way ANOVA was used, and pairwise comparisons were performed using the t-test. P <0.05, P <0.01 indicates that the difference is statistically significant.

[0282] 3.3 Test Results Table 48 Effects of potassium oxonate + yeast on a rat model of hyperuricemia complicated with acute gouty arthritis ( - (x±s, n=10)

[0283] Note: Compared with the normal control group: ## P <0.01; compared with the model control group: P <0.01.

[0284] Table 1 shows that before modeling, the serum uric acid levels in each group were uniform and without difference; after modeling (before drug administration), the serum uric acid levels in each group of rats increased significantly, showing a significant difference compared with the normal control group. P <0.01), and there was no difference between the groups; after 2 and 4 weeks of administration of the positive control drugs benzbromarone, Gout Relief, and Geshu Granules, the serum uric acid levels in all three dosage groups decreased significantly, and there were significant differences compared with the model control group. P <0.01).

[0285] Table 49 Effects of potassium oxonate + yeast on a rat model of hyperuricemia complicated with acute gouty arthritis ( - (x±s, n=10)

[0286] Note: Compared with the normal control group: ## P <0.01; compared with the model control group: P <0.01.

[0287] Table 49 shows that the serum CRP and Hcy levels in the model control group were significantly increased compared with those in the normal control group (P<0.01); the serum CRP and Hcy levels in the three dose groups of the positive control drugs benzbromarone, Tongfengshu, and Geshu granules were significantly decreased after 2 and 4 weeks of administration, and were significantly different from those in the model control group (P<0.05, P<0.01).

[0288] Table 50 Effects of potassium oxonate + yeast on a rat model of hyperuricemia complicated with acute gouty arthritis (drug administration for 2 weeks) - (x±s, n=10)

[0289] Note: Compared with the normal control group, ##P<0.01; compared with the model control group: P<0.01.

[0290] Table 51 Effects of potassium oxonate + yeast on a rat model of hyperuricemia complicated with acute gouty arthritis (drug administration for 4 weeks) - (x±s, n=10)

[0291] Note: Compared with the normal control group ## P <0.01; compared with the model control group: P <0.01.

[0292] Tables 49 and 50 show that the serum levels of TG, TC, CRE, UREA, and GLU in the model control group were significantly elevated compared with those in the normal control group (P<0.01). The three dose groups of the positive control drugs benzbromarone, Tongfengshu, and Geshu granules all significantly reduced the serum levels of TG, TC, CRE, UREA, and GLU in rats, showing significant differences compared with those in the model control group (P<0.01).

[0293] Table 52 Effects of potassium oxonate + yeast on a rat model of hyperuricemia complicated with acute gouty arthritis (drug administration for 4 weeks) - (x±s, n=10)

[0294] Note: Compared with the normal control group ## P <0.01; compared with the model control group: P <0.01.

[0295] Table 52 shows that the serum AST and ALT levels in the model control group were significantly elevated compared with the normal control group (P<0.01); the three dose groups of the positive control drug benzbromarone, Gout Relief, and Geshu Granules all significantly reduced the serum AST and ALT levels in rats, showing significant differences compared with the model control group. P <0.01).

[0296] Table 53 Effects of potassium oxonate + yeast on a rat model of hyperuricemia combined with acute gouty arthritis ( - (x±s, n=10)

[0297] Note: Compared with the normal control group: ##P<0.01; Compared with the model control group: P<0.01.

[0298] Table 53 shows that there was no difference in the volume of the right ankle joint among the groups before modeling; 3 days after modeling, the volume of the right ankle joint of the rats in the model control group was significantly increased, which was significantly different from that of the normal control group (P<0.01); the three dose groups of the positive control drugs benzbromarone, Tongfengshu, and Geshu granules could significantly reduce the volume of the right ankle joint of the rats after modeling, which was significantly different from that of the model control group (P<0.01).

[0299] Table 54 Effects of Geshu Granules on Inflammation Index Scores in Model Animals ( - (x±s, n=10)

[0300] Note: Compared with the normal control group, ## P <0.01; compared with the model control group, P <0.05; P <0.01.

[0301] Table 55 Effects of Geshu Granules on Functional Impairment Scores in Model Animals ( - (x±s, n=10)

[0302] Note: Compared with the normal control group, ## P<0.01 Compared with the model control group, Tables 54 and 55 show that at 12h, 24h, 48h, 72h, and 96h after modeling, the joint inflammation index and functional impairment scores of rats in the model control group were significantly increased, showing significant differences compared with the normal control group. P <0.01); The inflammation score and functional impairment score of the three dosage groups of Geshu granules all decreased to a certain extent. Among them, the inflammation index score of the high-dose group was significantly different from that of the model control group at 24h, 48h and 72h after modeling. P <0.01, the inflammatory index scores of the medium-dose group were significantly different from those of the model control group at 24h and 48h after modeling ( P <0.05, P <0.01, the low-dose group showed a significant difference in inflammation index score compared with the model control group 12 hours after modeling ( P <0.05).

[0303] 4. Conclusion Geshu granules have a good therapeutic effect on a rat model of hyperuricemia induced by potassium oxonate and yeast combined with acute gouty arthritis. They can reduce the levels of UA, TG, TC, CRE, UREA, GLU, AST and ALT in the blood; reduce CRP and Hcy levels; and reduce ankle swelling.

[0304] Example 7 Liver and kidney pathology examination report Submission Information Seventy HE-stained sections of rat kidney and liver tissue from each group in Example 6 were submitted for testing, with ten sections from each treatment group. The testing was conducted by the Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences.

[0305] Experimental Objective The study aimed to observe the lesions in the kidney and liver tissues and evaluate the effects of Geshu granules on the kidney and liver of rats with hyperuricemia and acute gouty arthritis.

[0306] Test methods HE staining was performed, and the pathological changes of rat kidney and liver tissues were observed under a light microscope and photographed.

[0307] For the histopathological grading of rat kidney tissue, please refer to Example 4.

[0308] The results of the light microscopic examination of the kidney pathology are shown in the figure. Figure 8 The details are as follows: 1. Normal control group: The rat kidneys were intact, the basement membrane was uniform, no mesangial cell proliferation was observed, the renal tubular epithelial cells were tightly arranged, the brush border was clear, there were no casts or crystal deposits, and there was no fibrous tissue proliferation or inflammatory cell infiltration in the interstitium.

[0309] 2. Model control group: The proximal tubular epithelial cells of rat kidneys showed mild or slight swelling, degeneration, and necrosis; brush border detachment; pink protein casts were present in the renal tubular lumen; medullary tubules were dilated; interstitial collagen fibers showed mild or slight proliferation; and myofibroblasts were activated. Significant differences were observed compared to the normal control group. P <0.01).

[0310] 3. Benzbromarone group, Gout Relief group, and high, medium, and low dose groups of Geshu granules: The extent and degree of renal tubular degeneration, necrosis, dilation, and interstitial collagen fiber proliferation in rats were significantly reduced compared with the model control group. P <0.05 or P <0.01), indicating a significant protective effect against kidney damage.

[0311] The degree of protection against kidney injury in rats by the above groups, from strongest to weakest, is as follows: high-dose Geshu granules group > medium-dose Geshu granules group > Tongfengshu group > benzbromarone group > low-dose Geshu granules group.

[0312] The results of the light microscopic examination of liver pathology are shown in Figure 9 The details are as follows: 1. Normal control group: The liver lobule structure was clear, the hepatocytes were of uniform size, no degeneration or necrosis was observed, no fibrosis or inflammatory cell infiltration was observed in the interstitium, and the tissue structure was normal.

[0313] 2. Model control group: Rat hepatocytes showed mild vacuolar degeneration (6 / 10), which was significantly different from the normal control group. P <0.01).

[0314] 3. Benzbromarone group, Gout Relief group, and high-dose Geshu granules group: The extent and degree of vacuolar degeneration in rat hepatocytes were significantly reduced compared with the model control group. P <0.05 or P <0.01), indicating a significant protective effect against hepatocellular damage.

[0315] 4. Medium-dose group of Geshu granules: The degree of vacuolar degeneration of rat hepatocytes was reduced compared with the model control group, and it had a slight protective effect on hepatocyte damage.

[0316] 5. Low-dose Gezhu granules group: The degree of vacuolar degeneration of rat hepatocytes showed a decreasing trend compared with the model control group. P <0.1), which has a slight protective effect against hepatocyte damage.

[0317] Table 56 Statistical analysis of pathological changes in rat renal tubular necrosis / dilation under microscopic observation

[0318] Note: Compared with the model group, P <0.05, P <0.01.

[0319] Table 57 Statistical analysis of pathological changes in rat renal interstitial collagen fiber proliferation under microscopic observation

[0320] Note: Compared with the model group, P <0.05, P <0.01.

[0321] Table 58 Statistical analysis of pathological changes in rat hepatocyte vacuolar degeneration observed under a microscope

[0322] Note: Compared with the model group, P <0.05, P <0.01.

[0323] in conclusion 1. Normal control group: The rat kidneys were intact, with a uniform basement membrane, no mesangial cell proliferation, tightly packed renal tubular epithelial cells, clear brush borders, no casts or crystal deposits, and no fibrous tissue proliferation or inflammatory cell infiltration in the interstitium; the liver lobules were clearly structured, the hepatocytes were uniform in size, no degeneration or necrosis was observed, and no fibrous tissue proliferation or inflammatory cell infiltration was observed in the interstitium, with normal tissue structure.

[0324] 2. Model control group: In rats, the proximal tubular epithelial cells showed mild or slight swelling, degeneration, and necrosis; brush border detachment; pink protein casts were present in the tubular lumen; medullary tubules were dilated; interstitial collagen fibers showed mild or slight proliferation; and myofibroblasts were activated. Hepatocytes showed mild vacuolar degeneration (6 / 10), significantly different from the normal group. This indicates that the model of hyperuricemia complicated with acute gouty nephropathy was successfully established.

[0325] 3. Benzbromarone group, Gout Relief group, and high, medium, and low dose groups of Geshu granules: The extent and degree of renal tubular degeneration, necrosis, dilation, and interstitial collagen fiber proliferation in rats were significantly reduced compared with the model control group, suggesting a significant protective effect against kidney damage. The order from strongest to weakest was: high dose group of Geshu granules > medium dose group of Geshu granules = Gout Relief group > benzbromarone group > low dose group of Geshu granules.

[0326] 4. Benzbromarone group, Gout Relief group, and high-dose Geshu granules group: The extent and degree of vacuolar degeneration of rat hepatocytes were significantly reduced compared with the model control group, suggesting a significant protective effect against hepatocyte damage.

[0327] 5. Medium-dose group of Geshu granules: The degree of vacuolar degeneration of rat hepatocytes was reduced compared with the model control group, and it had a slight protective effect on hepatocyte damage.

[0328] 6. Low-dose Geshu granules group: The degree of vacuolar degeneration of rat hepatocytes showed a decreasing trend compared with the model control group, and had a slight protective effect against hepatocyte damage.

[0329] Example 8 Rat joint pathology report Submission Information A total of 70 samples of HE-stained rat ankle joint tissue from each group in Example 6 were submitted for testing, with 10 samples from each treatment group. The testing institution was the Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences.

[0330] Experimental Objective To observe the lesions in the ankle joint tissue and evaluate the effect of Geshu granules on the ankle joint of rats with hyperuricemia and acute gouty arthritis.

[0331] Test methods HE staining was performed, and the pathological changes of rat ankle joint tissue were observed under a light microscope and photographed.

[0332] The histopathological grading of rat ankle joint tissue was based on the following criteria: ① synovial cell proliferation; ② interstitial inflammatory cell infiltration; ③ angiogenesis; and ④ the extent and degree of bone destruction. The lesions were classified into five grades. Statistical analysis: SPSS 20.0 statistical software was used for statistical analysis. When comparing sample means, the t-test of two independent sample means or one-way ANOVA was used for continuous data that met the requirements of normal distribution and homogeneity of variance. For those that did not meet the requirements, the nonparametric rank-sum test was used for statistical analysis of graded data. The p-value < 0.05 was used as the criterion for statistical significance.

[0333] The pathological light microscopy results are shown in Figure 10 The details are as follows: 1. Normal control group: The surface of the articular cartilage of rats was smooth, the chondrocytes were neatly arranged, the bone was normal, no congestion, edema and hyperplasia were observed in the synovium, no thickening of the joint capsule, and no fibrous tissue hyperplasia and inflammatory cell infiltration in the interstitium.

[0334] 2. Model control group: Microscopic examination revealed mild to severe proliferation of synovial cells in rat joints, with increased cell layers, interstitial congestion and edema accompanied by extensive inflammatory cell infiltration, including neutrophils, lymphocytes, and macrophages. Joint cavity narrowing was observed, along with significant inflammatory tissue necrosis and fibrinoid exudation. Vascular dilation and proliferation were observed, along with pannus formation, cartilage surface erosion, and significant subchondral bone destruction. Moderate to severe inflammatory cell infiltration was observed around the joint. Significant differences were observed compared to the normal control group. P <0.01).

[0335] 3. Benzbromarone group, Gout Relief group, and high, medium, and low dose groups of Geshu granules: The number and thickness of synovial cell layers in rat joints decreased, the inflammatory response in the synovium and surrounding interstitium was significantly reduced, the number of small blood vessels decreased, pannus significantly degenerated, the fibrotic area on the cartilage surface decreased, bone repair was significant, and the histopathological score was significantly lower than that of the model control group. P <0.05 or <0.01), which has a significant protective effect against gouty arthritis.

[0336] 4. The protective effect of each group against gouty arthritis in rats, from strongest to weakest, is as follows: high-dose Geshu granules group > benzbromarone group > medium-dose Geshu granules group > low-dose Geshu granules group > Tongfengshu group.

[0337] Table 59 Statistical analysis of pathological changes in synovial cell proliferation in rat ankle joint under microscopic observation

[0338] Note: Compared with the model group, P <0.05, P <0.01.

[0339] Table 60 Statistical analysis of pathological changes in interstitial inflammatory cell infiltration in rat ankle joint under microscopic observation

[0340] Note: Compared with the model group, P <0.05, P <0.01.

[0341] Table 61 Statistical analysis of pathological changes in angiogenesis in rat ankle joints under microscopic observation

[0342] Note: Compared with the model group, P <0.05, P <0.01.

[0343] Table 62 Statistical analysis of pathological changes in ankle joint bone destruction observed under a microscope in rats

[0344] Note: Compared with the model group, P <0.05, P <0.01.

[0345] in conclusion 1. Normal control group: The surface of the articular cartilage of rats was smooth, the chondrocytes were neatly arranged, the bone was normal, no congestion, edema and hyperplasia were observed in the synovium, no thickening of the joint capsule, and no fibrous tissue hyperplasia and inflammatory cell infiltration in the interstitium.

[0346] 2. Model control group: Microscopic examination revealed mild to severe proliferation of synovial cells in rat joints, with an increased number of cell layers, interstitial congestion and edema accompanied by a large number of inflammatory cell infiltrations, including neutrophils, lymphocytes and macrophages. The joint cavity was narrowed, with a large amount of inflammatory tissue necrosis and fibrinoid exudation. Vascular dilation and proliferation, pannus formation, cartilage surface erosion, and significant destruction of subchondral bone tissue were observed. Moderate or severe inflammatory cell infiltration was observed around the joints, which was significantly different from the normal control group, indicating that the model of hyperuricemia complicated with acute gouty arthritis was successfully established.

[0347] 3. Benzbromarone group, Gout Relief group, and high, medium, and low dose groups of Geshu granules: The number and thickness of synovial cells in rat joints decreased, the inflammatory response in the synovium and surrounding interstitium was significantly reduced, the number of small blood vessels decreased, the pannus was significantly degenerated, the fibrosis area on the cartilage surface decreased, bone repair was obvious, and the histopathological score was significantly lower than that of the model control group, showing a significant protective effect against gouty arthritis.

[0348] 4. The protective effect of each group against gouty arthritis in rats, from strongest to weakest, is as follows: high-dose Geshu granules group > benzbromarone group > medium-dose Geshu granules group > low-dose Geshu granules group > Tongfengshu group.

[0349] In summary, Geshu granules target multiple pathological damage points in hyperuricemia complicated with acute gouty arthritis, exhibiting rapid and sustained reduction in serum uric acid levels, relief of acute joint inflammation and swelling, control of systemic inflammatory response, regulation of lipid metabolism, and protection of renal function. This provides experimental data support for the clinical application of Geshu granules in the treatment of hyperuricemia, gouty arthritis, and related metabolic complications, and offers new ideas and methods for the treatment of hyperuricemia complicated with gouty arthritis using traditional Chinese medicine.

[0350] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A pharmaceutical composition, characterized in that, It is made from the following parts by weight of traditional Chinese medicine raw materials: 10-60 parts of kudzu root, 5-20 parts of atractylodes rhizome, 5-20 parts of phellodendron bark, 5-20 parts of smilax glabra rhizome, and 5-20 parts of achyranthes root.

2. The pharmaceutical composition according to claim 1, characterized in that, It is made from the following Chinese herbal raw materials in parts by weight: 20-40 parts of kudzu root, 7-15 parts of atractylodes rhizome, 8-13 parts of phellodendron bark, 10-17 parts of smilax glabra rhizome, and 6-15 parts of achyranthes bidentata root.

3. A method for preparing the pharmaceutical composition according to claim 1 or 2, characterized in that, Includes the following steps: Phellodendron bark and Smilax glabra were extracted separately or in combination by decoction to obtain a decoction. Atractylodes lancea was extracted by steam distillation to obtain Atractylodes lancea volatile oil, Atractylodes lancea water extract, and Atractylodes lancea residue; The Atractylodes lancea residue, Pueraria lobata and Achyranthes bidentata were extracted with alcohol, either separately or in combination, to obtain an alcohol extract. The decoction and Atractylodes lancea water extract were dehydrated to obtain the water extract; The volatile oil of Atractylodes lancea was encapsulated and then mixed with alcoholic and water extracts to obtain a pharmaceutical composition.

4. The preparation method according to claim 3, characterized in that, The extraction of Phellodendron bark and Smilax glabra was performed 2 to 3 times; the mass ratio of the total mass of Phellodendron bark and Smilax glabra to water was 1:6 to 8 during each extraction; and the extraction time was 1.5 to 2 hours.

5. The preparation method according to claim 3, characterized in that, During the steam distillation extraction, the mass ratio of Atractylodes lancea to water is 1~7.5~8.5; the extraction time is 5.5~6.5 hours. The method for encapsulating the volatile oil of Atractylodes lancea involves adding the volatile oil of Atractylodes lancea to an encapsulating excipient or an encapsulating excipient solution and encapsulating it using at least one of the following methods: colloid milling, ultrasonication, grinding, and water-saturated solution method.

6. The preparation method according to any one of claims 3 to 5, characterized in that, During the alcohol extraction, the volume concentration of the alcohol is 68%~72%; the alcohol includes ethanol; the alcohol extraction is performed 2~3 times; and the extraction time for each extraction is 1.5~2 hours.

7. A drug for preventing and treating hyperuricemia, associated liver and kidney damage, and gouty arthritis, characterized in that, The pharmaceutical composition and pharmaceutically acceptable excipients are obtained by the preparation method of any one of claims 1 to 2.

8. The drug according to claim 7, characterized in that, Pharmaceutically acceptable excipients include fillers and flavoring agents; The filler includes dextrin; the filler accounts for 20% to 40% of the mass of the pharmaceutical composition; The flavoring agent comprises stevioside and citric acid; the stevioside accounts for 0.8% to 1.2% of the mass of the pharmaceutical composition; and the citric acid accounts for 0.4% to 0.6% of the mass of the pharmaceutical composition.

9. The use of the pharmaceutical composition of claim 1 or 2 or the pharmaceutical composition obtained by the preparation method of any one of claims 3 to 6 in the preparation of a medicament for the prevention and / or treatment of at least one of the following diseases: hyperuricemia, hyperuricemia with liver and kidney damage, and hyperuricemia complicated with gouty arthritis.

10. The application according to claim 9, characterized in that, The complications of hyperuricemia combined with gouty arthritis also include kidney and / or liver damage.

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