Traditional Chinese medicine composition for treating animal gout and application thereof

Abstract

The invention discloses a traditional Chinese medicine composition for treating animal gout and application of the traditional Chinese medicine composition, and belongs to the technical field of animal medicine. The core active components of the traditional Chinese medicine composition comprise astilbin, tanshinone IIA and tetrahydrocurcumin, wherein the preferable mass ratio of all the components is 4: 1: 3. Pharmacodynamic studies show that the composition has a significant synergistic effect, can effectively reduce the blood uric acid level of animals with hyperuricemia, and synergistically inhibits the activity of xanthine oxidase (XOD). Compared with existing chemical drugs, the pharmaceutical composition disclosed by the invention has the advantages that through synergistic combination of the drugs, the toxic and side effects and the possibility of drug resistance are effectively reduced while the concentration of uric acid is efficiently reduced, a safe, efficient and economic brand-new scheme is provided for treating animal gout, and the pharmaceutical composition has important application value.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of animal medicine, and particularly relates to a traditional Chinese medicine combination drug for treating gout in animals and application thereof. BACKGROUND

[0002] Gout is a metabolic disease caused by hyperuricemia, which widely exists in humans and various animals. For example, in poultry, gout mainly manifests as deposition of urate in joints, kidneys and internal organs, thereby causing joint swelling, pain, kidney function damage, and seriously affecting the growth performance, egg production rate and economic benefits of poultry breeding.

[0003] At present, the treatment of gout in animals mainly relies on chemical drugs such as allopurinol and colchicine, which can alleviate symptoms by inhibiting uric acid synthesis or promoting uric acid excretion. However, the application of chemical drugs in animals has the problem of large toxic and side effects. In addition, long-term use of chemical drugs may lead to drug resistance and reduce the therapeutic effect. In recent years, traditional Chinese medicine has been widely concerned in the treatment of animal diseases due to its naturalness and low toxicity. Therefore, it is a problem to be solved to design a traditional Chinese medicine composition that can act on multiple targets and multiple pathways to efficiently reduce uric acid levels. SUMMARY

[0004] The purpose of the present application is to provide a traditional Chinese medicine combination drug for treating gout in animals and application thereof, so as to achieve the effect of efficiently reducing uric acid in poultry and other animals.

[0005] To achieve the above-mentioned purpose, the present application provides the following technical solutions: Firstly, the present application provides a traditional Chinese medicine combination drug for treating gout in animals, which is composed of a drug core active ingredient and a drug carrier. The drug core active ingredient includes astilbin, tanshinone ⅡA and tetrahydrocurcumin.

[0006] Preferably, in the traditional Chinese medicine combination drug, the mass ratio of astilbin, tanshinone ⅡA and tetrahydrocurcumin is 3-5:0.5-1.5:2.5-3.5.

[0007] Preferably, in the traditional Chinese medicine combination drug, the mass ratio of astilbin, tanshinone ⅡA and tetrahydrocurcumin is 4:1:3.

[0008] Preferably, in 100 mL of the traditional Chinese medicine combination drug, the content of the core active ingredient is: astilbin 0.4 g, tanshinone ⅡA 0.1 g, tetrahydrocurcumin 0.3 g, and the rest is a drug carrier. The drug carrier is a drug carrier composed of 5% (v / v) ethanol and 95% (v / v) physiological saline.

[0009] Preferably, the gout is hyperuricemic gout.

[0010] Secondly, the present invention provides an application of a traditional Chinese medicine composition in the preparation of a drug for treating gout in animals, characterized in that the traditional Chinese medicine composition is composed of astilbene, tanshinone IIA and tetrahydrocurcumin.

[0011] Preferably, the mass ratio of astilbene, tanshinone IIA and tetrahydrocurcumin in the traditional Chinese medicine composition is 3-5:0.5-1.5:2.5-3.5.

[0012] Preferably, in every 100 mL of the drug, the content of astilbene is 0.4 g, the content of tanshinone IIA is 0.1 g, and the content of tetrahydrocurcumin is 0.3 g.

[0013] Preferably, the gout is hyperuricemic gout.

[0014] In addition, the present invention provides an application of a traditional Chinese medicine composition in the preparation of a drug for reducing uric acid levels in animals, wherein each 100 mL of the drug contains 0.4 g of astilbene, 0.1 g of tanshinone IIA, and 0.3 g of tetrahydrocurcumin.

[0015] The beneficial effects of this invention are as follows: This invention provides a traditional Chinese medicine composition for treating gout in animals. By scientifically proportioning astilbin, tanshinone IIA, and tetrahydrocurcumin, a significant synergistic effect is achieved. Pharmacodynamic studies show that this composition exhibits excellent efficacy in reducing serum uric acid levels in hyperuricemic mice, with its actual uric acid-lowering effect significantly superior to that of any single component used alone or in combination (i.e., the synergistic effect conforms to "1+1+1>3"). This synergistic effect is not only reflected in the significant decrease in serum uric acid levels but also in its synergistic inhibition of the key enzyme xanthine oxidase (XOD) activity, revealing the unique advantage of this composition's multi-target, multi-pathway synergistic action. Compared with existing chemical drugs, the traditional Chinese medicine composition of this invention has advantages such as being natural, having low toxicity and side effects, and being less likely to induce drug resistance. It provides a safe, efficient, and economical new solution for treating gout in animals, and has significant practical significance and application value for ensuring the healthy development of the poultry farming industry. Attached Figure Description

[0016] Figure 1 This is a graph showing the effects of the herbal composition of this invention and various control drugs on serum uric acid levels in a mouse model of hyperuricemia and gout. Figure 2This is a graph showing the effects of the herbal composition of this invention and various control drugs on the xanthine oxidase (XOD) activity in a mouse model of hyperuricemia and gout. Figure 3 The figure shows the effect of the traditional Chinese medicine compositions B, C and D of the present invention on the serum uric acid level in a mouse model of hyperuricemia and gout. Detailed Implementation

[0017] Drug raw materials: Astilbin, with the molecular formula C 21 H 22 O 11 Its CAS number is 29838-67-3, and its structure is as follows:

[0018] Tanshinone IIA, its molecular formula is C 19 H 18 O3, with CAS number 568-72-9, has the following structural formula:

[0019] Tetrahydrocurcumin has the molecular formula C 21 H 24 O6, with CAS number 36062-04-1, has the following structural formula: .

[0020] Example 1 A traditional Chinese medicine composition A for treating gout in animals The composition of each 100mL of traditional Chinese medicine composition A is as follows:

[0021] The preparation method of the traditional Chinese medicine composition A is as follows: (1) Weigh out 0.4 g of astilbene, 0.1 g of tanshinone IIA and 0.3 g of tetrahydrocurcumin and place them in a clean preparation container; (2) Add a mixed solvent consisting of 5% ethanol and 95% physiological saline, and stir thoroughly until all components are completely dissolved; (3) Add solvent to a total volume of 100 mL and continue stirring to make the solution homogeneous; (4) The solution was aseptically filtered through a 0.22 μm microporous membrane to remove microorganisms and insoluble particles; (5) Dispense the contents into light-proof containers under sterile conditions, seal them, and store them in the refrigerator (2-8℃) to obtain the traditional Chinese medicine composition A.

[0022] Example 2 A traditional Chinese medicine composition B for treating gout in animals The composition of each 100mL of traditional Chinese medicine composition B is as follows:

[0023] The preparation method of this traditional Chinese medicine composition B is as follows: (1) Weigh out 0.3 g of astilbene, 0.15 g of tanshinone IIA and 0.35 g of tetrahydrocurcumin and place them in a clean preparation container; (2) Add a mixed solvent consisting of 5% ethanol and 95% physiological saline, and stir thoroughly until all components are completely dissolved; (3) Add solvent to a total volume of 100 mL and continue stirring to make the solution homogeneous; (4) The solution was aseptically filtered through a 0.22 μm microporous membrane to remove microorganisms and insoluble particles; (5) Dispense the contents into light-proof containers under sterile conditions, seal them, and store them in the refrigerator (2-8℃) to obtain the traditional Chinese medicine composition B.

[0024] Example 3 A traditional Chinese medicine composition for treating gout in animals C The composition of each 100mL of traditional Chinese medicine composition C is as follows:

[0025] The preparation method of the traditional Chinese medicine composition C is as follows: (1) Weigh out 0.5 g of astilbene, 0.05 g of tanshinone IIA and 0.25 g of tetrahydrocurcumin and place them in a clean preparation container; (2) Add a mixed solvent consisting of 5% ethanol and 95% physiological saline, and stir thoroughly until all components are completely dissolved; (3) Add solvent to a total volume of 100 mL and continue stirring to make the solution homogeneous; (4) The solution was aseptically filtered through a 0.22 μm microporous membrane to remove microorganisms and insoluble particles; (5) Dispense the contents into light-proof containers under sterile conditions, seal them, and store them in the refrigerator (2-8℃) to obtain the traditional Chinese medicine composition C.

[0026] Example 4 A traditional Chinese medicine composition for treating gout in animals (D) The composition of each 100mL of traditional Chinese medicine composition D is as follows:

[0027] (1) Weigh out 0.36 g of astilbene, 0.12 g of tanshinone IIA and 0.32 g of tetrahydrocurcumin and place them in a clean preparation container; (2) Add a mixed solvent consisting of 5% ethanol and 95% physiological saline, and stir thoroughly until all components are completely dissolved; (3) Add solvent to a total volume of 100 mL and continue stirring to make the solution homogeneous; (4) The solution was aseptically filtered through a 0.22 μm microporous membrane to remove microorganisms and insoluble particles; (5) Dispense the contents into light-proof containers under sterile conditions, seal them, and store them in the refrigerator (2-8℃) to obtain the traditional Chinese medicine composition D.

[0028] Comparative Example 1 Comparison drug A Composition: 100 mL of the drug was prepared using 5% ethanol (v / v) + 95% physiological saline (0.9% NaCl) as a carrier, containing 0.4 g of astilbin.

[0029] Comparative Example 2 Comparison drug B Composition: 100 mL of the drug was prepared using 5% ethanol (v / v) + 95% physiological saline (0.9% NaCl) as a carrier, containing 0.1 g of tanshinone IIA.

[0030] Comparative Example 3 Comparison drug C Composition: 100 mL of the drug was prepared using 5% ethanol (v / v) + 95% physiological saline (0.9% NaCl) as a carrier, containing 0.3 g of tetrahydrocurcumin.

[0031] Comparative Example 4 Comparison drug D Composition: 100 mL of the drug was prepared using 5% ethanol (v / v) + 95% physiological saline (0.9% NaCl) as a carrier, containing 0.4 g of astilbene and 0.1 g of tanshinone IIA.

[0032] Comparative Example 5 Comparison drug E Composition: 100 mL of the drug was prepared using 5% ethanol (v / v) + 95% physiological saline (0.9% NaCl) as a carrier, containing 0.4 g of astilbin and 0.3 g of tetrahydrocurcumin.

[0033] Comparative Example 6 Comparison drug F Composition: 100 mL of the drug was prepared using 5% ethanol (v / v) + 95% physiological saline (0.9% NaCl) as a carrier, containing 0.1 g of tanshinone IIA and 0.3 g of tetrahydrocurcumin.

[0034] Example 5 The effect of the traditional Chinese medicine composition A of the present invention on serum uric acid levels in a mouse model of hyperuricemia was investigated. (1) Model building and grouping Animal selection: Male C57BL / 6 mice (weighing 20-25 g) aged 6-8 weeks were selected and purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0035] Housing environment: Standard SPF grade animal room, temperature 22±2℃, relative humidity 50-60%, 12-hour light-dark cycle (lighting time 8:00-20:00).

[0036] Feed and water: Standard purine-free mouse feed and distilled water were provided freely.

[0037] Adaptation period: 1 week, during which time animal behavior and health status (such as weight, appetite, and activity level) are observed, and abnormal individuals are excluded.

[0038] Reagent preparation: Hypoxanthine (HX): purity >98%, dissolved in 0.9% physiological saline, concentration 200 mg / mL.

[0039] Potassium oxazine (PO): Purity >98%, soluble in 0.9% physiological saline, concentration 100 mg / mL.

[0040] Induction methods: After weighing the mice, they were injected intraperitoneally with HX 200 mg / kg + PO 100 mg / kg. The mice were observed for 10 minutes after the injection to ensure there were no abnormal reactions. After 7 consecutive days of injection, blood was collected from the tail vein and the serum uric acid concentration was measured using an ELISA kit. If the serum uric acid concentration was >200 μmol / L, the modeling was successful and a hyperuricemia gout model was obtained. Normal control group: Male C57BL / 6 mice were injected with an equal volume of physiological saline at the same time.

[0041] Experimental Groups: normal control group Intervention subjects: Normal mice (without a hyperuricemia model) Intervention method: Gavage (drug carrier: 5% ethanol (v / v) + 95% saline (0.9% NaCl)); Model group Intervention subjects: Hyperuricemia and gout model mice (after successful modeling) Intervention method: Gavage (using the same drug carrier as the normal control group to exclude the influence of the carrier itself on the experimental results); Astilbene group Intervention subjects: Mice with hyperuricemia and gout Intervention method: Gavage (comparison drug A); Tanshinone IIA group Intervention subjects: Mice with hyperuricemia and gout Intervention method: Gavage (comparison drug B); Tetrahydrocurcumin group Intervention subjects: Mice with hyperuricemia and gout Intervention method: Gavage (comparison drug C); Astilbene + Tanshinone IIA group Intervention subjects: Mice with hyperuricemia and gout Intervention method: Gavage (comparison drug D); Astilbene + Tetrahydrocurcumin group Intervention subjects: Mice with hyperuricemia and gout Intervention method: Gavage (comparison drug E); Tanshinone IIA + Tetrahydrocurcumin group Intervention subjects: Mice with hyperuricemia and gout Intervention method: Gavage (comparison drug F); Group A of Traditional Chinese Medicine Compositions Intervention subjects: Mice with hyperuricemia and gout Intervention method: Gavage (Chinese herbal composition A).

[0042] Dosage: 200 μL / 20g body weight (i.e., 10 μL per 1g body weight); Dosage time: 9:00 AM daily; Dosage period: 14 consecutive days by gavage; Sample size: 10 animals per group.

[0043] After treatment, tail vein blood was collected, and serum uric acid concentration was measured using an ELISA kit. During the experiment, no lethargy, loss of appetite, or disheveled fur was observed in the mice, indicating that the composition did not cause significant harm to the survival and health of the mice at the experimental dose.

[0044] Table 1. Effects of different drug treatments on serum uric acid levels in a mouse model of hyperuricemia.

[0045] Note: Actual treatment effect (%) = [(Model group serum uric acid level - Treatment group serum uric acid level) / (Model group serum uric acid level - Normal control group serum uric acid level)] × 100%; The formula for calculating the expected effect of the two drugs in combination is: Eab = Ea + Eb - Ea × Eb; The formula for calculating the expected effect of the three drugs in combination is: Eabc=Ea+Eb+Ec-Ea×Eb-Ea×Ec-Eb×Ec-Ea×Eb×Ec.

[0046] From Table 1 and Figure 1 The results showed that the serum uric acid level of the normal control group mice was 80.43±2.93 μmol / L, which is consistent with the normal physiological state; the serum uric acid level of the model group was significantly increased to 247.96±5.42 μmol / L, indicating that the hyperuricemia model was successfully established, and the serum uric acid level was 167.53 μmol / L higher than that of the normal control group.

[0047] The serum uric acid levels in the astilbene group, tanshinone IIA group, and tetrahydrocurcumin group were lower than those in the model group, but the overall reduction effect was weak.

[0048] The serum uric acid level in the astilbene + tanshinone IIA group decreased to 166.32±4.24 μmol / L, with an actual therapeutic effect of 48.73%, slightly higher than the expected therapeutic effect of 46.84%. The serum uric acid level in the astilbene + tetrahydrocurcumin group decreased to 161.51±5.29 μmol / L, with an actual therapeutic effect of 51.60%, slightly higher than the expected therapeutic effect of 49.54%. The serum uric acid level in the tanshinone IIA + tetrahydrocurcumin group decreased to 172.49±4.56 μmol / L. Although slightly lower than the other combination groups, it was significantly better than the single-drug groups, with an actual therapeutic effect of 45.04%, slightly higher than the expected therapeutic effect of 44.19%. The results above show that when astilbene, tanshinone IIA, and tetrahydrocurcumin are used in combination, although the effect is better than that of using a single drug, the actual effect produced by the combination is only slightly higher than expected. This indicates that the components only exert a slight synergistic effect and cannot produce a synergistic therapeutic effect on the reduction of blood uric acid.

[0049] The serum uric acid level in the traditional Chinese medicine composition group decreased to 104.60±4.66 μmol / L, a significant reduction approaching the level of the normal control group, demonstrating the strongest uric acid-lowering effect. Its actual therapeutic effect was 85.57%, far exceeding the expected therapeutic effect of 57.41%. The significant difference between the actual and expected effects indicates that the traditional Chinese medicine composition provided by this invention has a significant synergistic effect. This may be due to the synergistic effect of multiple components and multiple targets, leading to a significant reduction in serum uric acid levels in gout mice, thereby effectively treating hyperuricemia.

[0050] Example 6 The effect of the traditional Chinese medicine composition A of the present invention on xanthine oxidase activity in a mouse model of hyperuricemia and gout was investigated. (1) After blood was collected from each group of mice, the mice were euthanized and dissected to obtain liver tissue from each group of mice.

[0051] (2) The liver tissue was washed with pre-cooled phosphate buffer (PBS, pH 7.4), weighed, and then added to the buffer at a ratio of 1:9 (i.e., 1 g of tissue to 9 mL of buffer). The tissue was homogenized in an ice bath using a homogenizer.

[0052] (3) The homogenate was centrifuged at 3500 rpm for 15 minutes at 4°C, and the supernatant was used as XOD enzyme solution.

[0053] (4) The xanthine oxidase activity of mice in each group was detected using the XOD activity assay kit.

[0054] Table 2. Effects of different drug treatments on xanthine oxidase (XOD) activity in a mouse model of hyperuricemia and gout.

[0055] Note: Actual inhibition rate (%) = [(Model group XOD activity - Treatment group XOD activity) / (Model group XOD activity - Normal control group XOD activity)] × 100%; The formula for calculating the expected effect of the two drugs in combination is: Eab = Ea + Eb - Ea × Eb; The formula for calculating the expected effect of the three drugs in combination is: Eabc=Ea+Eb+Ec-Ea×Eb-Ea×Ec-Eb×Ec-Ea×Eb×Ec.

[0056] From Table 2 and Figure 2 The results show that, consistent with the results of blood uric acid testing, astilbene, tanshinone IIA, and tetrahydrocurcumin, when used alone, can reduce XOD activity, but the effect is relatively limited.

[0057] The combined use of any two of astilbene, tanshinone IIA, and tetrahydrocurcumin was more effective than any single drug in reducing XOD activity. However, the actual inhibition rate of the astilbene + tanshinone IIA group was 53.02%, slightly higher than the expected inhibition rate of 51.85%; the actual inhibition rate of the astilbene + tetrahydrocurcumin group was 54.85%, slightly higher than the expected inhibition rate of 53.71%; and the actual inhibition rate of the tanshinone IIA + tetrahydrocurcumin group was 46.70%, slightly lower than the expected inhibition rate of 48.01%. These results indicate that the combined use of astilbene, tanshinone IIA, and tetrahydrocurcumin in pairs does not have a synergistic effect in reducing XOD activity.

[0058] The XOD activity of the herbal composition group was significantly reduced, far superior to other groups, and close to the level of the normal control group, demonstrating a remarkable XOD inhibition effect. Furthermore, its actual inhibition effect (89.84%) far exceeded the expected inhibition effect (65.96%), indicating that the herbal composition A of the present invention can synergistically inhibit XOD activity, thereby reducing uric acid levels and achieving effective treatment of hyperuricemia and gout.

[0059] Example 7 The therapeutic effect of the traditional Chinese medicine composition BD of the present invention on hyperuricemia and gout was tested. (1) The model construction is the same as in Example 5, and the groups are as follows: normal control group Intervention subjects: Normal mice (without a hyperuricemia model) Intervention method: Gavage (drug carrier: 5% ethanol (v / v) + 95% saline (0.9% NaCl)); Model group Intervention subjects: Hyperuricemia and gout model mice (after successful modeling) Intervention method: Gavage (using the same drug carrier as the normal control group to exclude the influence of the carrier itself on the experimental results); Group B of Traditional Chinese Medicine Compositions Intervention subjects: Hyperuricemia and gout model mice (after successful modeling, the model construction method is the same as in Example 5). Intervention method: Gavage (Chinese herbal composition B); Group C of traditional Chinese medicine compositions Intervention subjects: Mice with hyperuricemia and gout Intervention method: Gavage (Chinese herbal composition C); Group D of Chinese herbal composition Intervention subjects: Mice with hyperuricemia and gout Intervention method: Gavage (Chinese herbal composition D).

[0060] Dosage: 200 μL / 20g body weight (i.e., 10 μL per 1g body weight); Dosage time: 9:00 AM daily; Dosage period: 14 consecutive days by gavage; Sample size: 10 animals per group.

[0061] After treatment, blood was collected from the tail vein, and the blood uric acid concentration was measured using an ELISA kit.

[0062] Table 3. Effects of different traditional Chinese medicine compositions on serum uric acid levels in a mouse model of hyperuricemia and gout.

[0063] From Table 3 and Figure 3The results show that the serum uric acid levels in the traditional Chinese medicine composition groups B, C, and D were significantly lower than those in the model group, indicating that the traditional Chinese medicine compositions B, C, and D prepared in this invention have significant therapeutic effects on hyperuricemic gout mice.

Claims

1. A traditional Chinese medicine combination for treating gout in animals, characterized in that, The traditional Chinese medicine combination drug consists of core active ingredients and a drug carrier; The core active ingredients of the drug include astilbin, tanshinone IIA, and tetrahydrocurcumin.

2. The traditional Chinese medicine combination drug according to claim 1, characterized in that, According to the mass ratio, the mass ratio of astilbin, tanshinone IIA and tetrahydrocurcumin in the aforementioned traditional Chinese medicine combination is 3-5:0.5-1.5:2.5-3.

5.

3. The traditional Chinese medicine combination drug according to claim 1, characterized in that, Based on the mass ratio, the mass ratio of astilbene, tanshinone IIA and tetrahydrocurcumin in the aforementioned traditional Chinese medicine combination is 4:1:

3.

4. The traditional Chinese medicine combination drug according to claim 3, characterized in that, In every 100 mL of the aforementioned traditional Chinese medicine combination, the content of the core active ingredients is: astilbin 0.4 g, tanshinone IIA 0.1 g, tetrahydrocurcumin 0.3 g, and the remainder is a drug carrier; The drug carrier is a combination of 5% (v / v) ethanol and 95% (v / v) saline.

5. The traditional Chinese medicine combination drug according to claim 4, characterized in that, The gout mentioned refers to gout caused by hyperuricemia.

6. The application of a traditional Chinese medicine composition in the preparation of a drug for treating gout in animals, characterized in that, The traditional Chinese medicine composition consists of astilbene, tanshinone IIA, and tetrahydrocurcumin.

7. The application according to claim 6, characterized in that, According to the mass ratio, the mass ratio of astilbin, tanshinone IIA and tetrahydrocurcumin in the traditional Chinese medicine composition is 3-5:0.5-1.5:2.5-3.

5.

8. The application according to claim 7, characterized in that, In every 100 mL of the drug, the content of astilbene is 0.4 g, the content of tanshinone IIA is 0.1 g, and the content of tetrahydrocurcumin is 0.3 g.

9. The application according to claim 8, characterized in that, The gout mentioned refers to gout caused by hyperuricemia.

10. The application of a traditional Chinese medicine composition in the preparation of a drug for lowering uric acid levels in animals, characterized in that, In every 100 mL of the drug, the content of astilbene is 0.4 g, the content of tanshinone IIA is 0.1 g, and the content of tetrahydrocurcumin is 0.3 g.

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