Thaflavin composition for preventing and treating hyperuricemia as well as preparation method and application of theaflavin composition

Abstract

The invention provides a theaflavin composition for preventing and treating hyperuricemia as well as a preparation method and application thereof, and relates to the technical field of medicines. The theaflavin composition comprises theaflavin, a chicory extract, a hovenia dulcis thunb extract, a semen plantaginis extract, a poria cocos extract, a pericarpium citri reticulatae extract and a corn stigma extract. The theaflavin, the cichorium intybus extract, the hovenia dulcis thunb extract and the semen plantaginis extract are used for inhibiting the activity of xanthine oxidase and regulating the function of uric acid transporter to realize bidirectional regulation of uric acid generation and excretion aiming at the diseases of excessive uric acid generation and excretion reduction; the poria cocos extract and the pericarpium citri reticulatae extract fundamentally improve internal environment disorder through transportation and transformation functions of the spleen and the stomach, reduce generation of phlegm dampness and stasis and create good body conditions for uric acid metabolism; the corn stigma extract guides all the medicines to directly reach the disease site, enhances the concentration and action efficiency of the medicines in excretion organs, accurately positions the urinary system, and forms a multi-target all-directional synergistic treatment system of treating symptoms, treating root causes and inducing meridians.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a theaflavin composition for preventing and treating hyperuricemia, and a preparation method and application thereof. Background Art

[0002] Hyperuricemia (HUA), a metabolic syndrome caused by purine metabolism disorders, has become a global health concern. Hyperuricemia is diagnosed when the blood uric acid level exceeds 420 μmol / L on two separate days. In recent years, its incidence has increased significantly, with a younger patient population, placing a heavy burden on both society and individuals. The dangers of hyperuricemia should not be underestimated. On the one hand, it is a significant risk factor for gout, with approximately 5%-12% of patients eventually developing gout, resulting in symptoms such as red, swollen, hot, and painful joints, tophi, and proteinuria, severely impacting joint function and quality of life. On the other hand, hyperuricemia is closely associated with numerous chronic diseases and is an independent risk factor for the development of metabolic syndrome, type 2 diabetes, and cardiovascular disease. It can trigger kidney problems such as gouty nephropathy, urinary stones, and even acute renal failure. It can also affect the eyes, causing blepharitis, tophi, and conjunctivitis. Currently, treatment options for hyperuricemia are limited and have numerous shortcomings. While drug therapy can control blood uric acid levels to a certain extent, it has significant drawbacks. For example, allopurinol can cause severe allergic reactions in some patients, and its use is restricted in those with renal insufficiency, requiring dosage adjustments. Febuxostat is relatively expensive, placing financial strain on patients with long-term use. Benzbromarone can cause adverse reactions such as gastrointestinal discomfort and liver damage, and is contraindicated in those with urinary stones or severe renal impairment. Therefore, the development of a safe and effective product for the prevention and treatment of hyperuricemia is urgent. Summary of the Invention

[0003] The object of the present invention is to provide a theaflavin composition for preventing and treating hyperuricemia, and a preparation method and application thereof. The theaflavin composition provided by the present invention has a significant effect in preventing and treating hyperuricemia.

[0004] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions: The present invention provides a theaflavins composition for preventing and treating hyperuricemia. The composition comprises the following raw materials in parts by weight: 15-20 parts of theaflavins, 15-20 parts of chicory extract, 8-12 parts of Hovenia dulcis fruit extract, 10-15 parts of Plantago seed extract, 10-15 parts of Poria cocos extract, 5-8 parts of tangerine peel extract and 3-5 parts of corn silk extract.

[0005] More preferably, the chicory extract is obtained by liquid anaerobic fermentation of composite microorganisms.

[0006] Preferably, the composite microorganism consists of Lactobacillus plantarum grx03 and Bifidobacterium animalis subspecies lactis BX-245.

[0007] Preferably, the Hovenia dulcis fruit extract is obtained by ethanol ultrasonic extraction.

[0008] Preferably, the Poria cocos extract is obtained by complex enzyme hydrolysis and ethanol precipitation.

[0009] Preferably, the complex enzyme consists of cellulase, xylanase and neutral protease.

[0010] Preferably, the tangerine peel extract is subjected to distillation to obtain volatile oil, the distilled medicinal residue is subjected to ethanol extraction to obtain an ethanol extract, and the volatile oil and the ethanol extract are mixed to obtain the tangerine peel extract.

[0011] The present invention also provides a method for preparing the above-mentioned theaflavins composition, comprising: mixing hydroxypropyl-β-cyclodextrin with water to obtain a hydroxypropyl-β-cyclodextrin solution, adding theaflavins, chicory extract, Hovenia dulcis fruit extract, Plantago seed extract, Poria cocos extract, Tangerine peel extract and Corn silk extract, performing ultrasonic inclusion complexation, and spray drying to obtain the theaflavins composition.

[0012] Preferably, the volume ratio of the total weight of theaflavins, chicory extract, Hovenia dulcis fruit extract, Plantago seed extract, Poria cocos extract, Tangerine peel extract and Corn silk extract to the hydroxypropyl-β-cyclodextrin solution is 1:12-18 g / mL.

[0013] The present invention also provides the use of the theaflavin composition in preparing a drug for treating hyperuricemia.

[0014] Compared with the prior art, the present invention has the following beneficial effects: The present invention provides a theaflavins composition for preventing and treating hyperuricemia, comprising theaflavins, chicory extract, Hovenia dulcis fruit extract, Plantago seed extract, Poria cocos extract, Tangerine peel extract, and Corn silk extract. The theaflavins, chicory extract, Hovenia dulcis fruit extract, and Plantago seed extract target the condition of excessive uric acid production and decreased excretion by inhibiting xanthine oxidase activity and regulating uric acid transporter function, thereby achieving bidirectional regulation of uric acid production and excretion. Poria cocos extract and Tangerine peel extract, through the spleen and stomach's transportation and transformation functions, fundamentally improve internal environmental disorders, reduce the formation of phlegm, dampness, and stasis, and create favorable conditions for uric acid metabolism. Corn silk extract guides various drugs directly to the site of disease, enhancing their concentration and efficacy in excretory organs and precisely targeting the urinary system, forming a multi-target, comprehensive, synergistic treatment system that addresses both the symptoms and the root cause, while also guiding the flow of meridians. DETAILED DESCRIPTION

[0015] The present invention provides a theaflavins composition for preventing and treating hyperuricemia. The composition comprises the following raw materials in parts by weight: 15-20 parts of theaflavins, 15-20 parts of chicory extract, 8-12 parts of Hovenia dulcis fruit extract, 10-15 parts of Plantago seed extract, 10-15 parts of Poria cocos extract, 5-8 parts of tangerine peel extract and 3-5 parts of corn silk extract.

[0016] The theaflavins composition for preventing and treating hyperuricemia of the present invention preferably comprises the following raw materials in parts by weight: 18 parts of theaflavins, 18 parts of chicory extract, 10 parts of Hovenia dulcis fruit extract, 12 parts of Plantago seed extract, 12 parts of Poria cocos extract, 7 parts of tangerine peel extract and 4 parts of corn silk extract.

[0017] From the perspective of traditional Chinese medicine, theaflavins inhibit xanthine oxidase activity and reduce uric acid production. Chicory clears heat, promotes diuresis, and dissolves urate crystals, directly targeting the core pathogenesis of hyperuricemia "damp-heat and blood stasis", inhibiting uric acid reabsorption and promoting uric acid excretion. The combination of theaflavins and chicory forms a dual pathway of "excretion + inhibition of production", which is the main drug; Plantago seed clears heat, promotes diuresis and relieves stranguria, increases urine volume to promote uric acid excretion, and guides damp heat out of the urine. Hovenia dulcis clears damp heat, assists the main drug in excreting uric acid, and harmonizes the spleen and stomach, which is the minister drug; Poria Poria strengthens the spleen and eliminates dampness, improves the root cause of metabolic disorders caused by spleen deficiency and dampness, solves the pathological basis of "endogenous dampness and turbidity", and indirectly reduces uric acid production by enhancing the transportation and transformation of the spleen and stomach. Tangerine peel regulates qi and strengthens the spleen, dries dampness and resolves phlegm, and prevents diuretic drugs (such as Plantago seed) from damaging qi and consuming yin. At the same time, it harmonizes gastrointestinal function, promotes drug absorption, and can regulate gastrointestinal motility, relieving side effects such as abdominal distension and loose stools that may be caused by diuretic drugs; corn silk guides the drugs directly to the kidney and bladder meridians, enhancing the targeting of diuresis and uric acid excretion, while slightly lowering blood pressure and improving renal microcirculation.

[0018] From the perspective of modern pharmaceutical theory, theaflavins, chicory extract, Hovenia dulcis extract and Plantago seed extract target the symptoms of excessive uric acid production and reduced excretion by inhibiting the activity of xanthine oxidase and regulating the function of uric acid transporters, thereby achieving bidirectional regulation of uric acid production and excretion; Poria cocos extract and Tangerine peel extract improve the internal environment disorders from the root through the spleen and stomach transportation and transformation functions, reduce the formation of phlegm, dampness and blood stasis, and create good conditions for uric acid metabolism; Corn silk extract guides the various drugs directly to the site of the disease, enhances the concentration and efficiency of the drugs in the excretory organs, and accurately locates the urinary system, forming a multi-target, comprehensive synergistic treatment system that treats the symptoms, cures the root cause, and guides the meridians.

[0019] The chicory extract of the present invention is preferably obtained by liquid anaerobic fermentation of composite microorganisms: the chicory stems and leaves are crushed, mixed with water, inoculated with composite microorganisms, and fermented at 35-40°C and pH 6-7 for 60-72 hours; more preferably, the chicory stems and leaves are crushed, mixed with water, inoculated with composite microorganisms, and fermented at 37°C and pH 6.5 for 66 hours. The plant lactobacillus grx03 was purchased from the General Microorganism Center of the China National Committee for the Collection of Microorganisms, with a deposit number of CGMCC No. 21268, and is disclosed in CN112795500A; the animal Bifidobacterium lactis subspecies BX-245 was purchased from the General Microorganism Center of the China National Committee for the Collection of Microorganisms, with a deposit number of CGMCC No. 26973, and is disclosed in CN116769654A. The number of plant lactobacillus grx03 in the composite microorganism is (4-7)×10 9 / g, and the bacterial count of animal Bifidobacterium lactis subsp. lactis BX-245 was (5-10)×10 8 The invention adopts specific microorganisms to ferment chicory, and the chicory extract obtained by water extraction and alcohol precipitation has a better effect of lowering uric acid.

[0020] The Hovenia dulcis fruit extract of the present invention is preferably obtained by ultrasonic extraction with ethanol: the Hovenia dulcis fruit is crushed, mixed with a 55%-65% ethanol solution, and ultrasonically extracted one to three times at a temperature of 40-50°C and a power of 250-300W, each for 8-15 minutes. More preferably, the Hovenia dulcis fruit is crushed, mixed with a 60% ethanol solution, and ultrasonically extracted two times at a temperature of 45°C and a power of 280W, each for 10 minutes. The Hovenia dulcis fruit extract prepared using the present invention inhibits uric acid synthesis, enhances renal excretion, and alleviates oxidative stress and liver and kidney damage caused by hyperuricemia.

[0021] The Poria extract of the present invention is obtained by complex enzyme enzymolysis and ethanol precipitation: the Poria sclerotium is crushed, mixed with water, and a complex enzyme is added to ultrasonically extract at a temperature of 50-55 ° C, a pH value of 5-5.5, and a power of 350-450W for 18-25 minutes, the enzyme is killed, filtered, the filtrate is concentrated, an ethanol solution is added for precipitation, centrifuged, and the precipitate is dried to obtain the Poria extract; the Poria sclerotium is crushed, mixed with water, a complex enzyme is added to ultrasonically extract at a temperature of 52 ° C, a pH value of 5.2, and a power of 400W for 20 minutes, the enzyme is killed, filtered, the filtrate is concentrated, an ethanol solution is added for precipitation, centrifuged, and the precipitate is dried to obtain the Poria extract. The complex enzyme is preferably composed of cellulase, xylanase, and neutral protease in a mass ratio of 5-7:2-4:0.8-2, more preferably 6:3:1. The Poria extract prepared by the present invention can improve the body's metabolic environment, reduce the generation of pathological products such as phlegm, dampness, and blood stasis, and provide favorable conditions for the normal metabolism of uric acid (such as the balance between synthesis and excretion). When used in combination with the tangerine peel extract, it can regulate gastrointestinal function and promote drug absorption.

[0022] The tangerine peel extract of the present invention preferably comprises: crushing the tangerine peel, mixing it with water and soaking it for 30-50 minutes, distilling it at 102-107° C. for 1.5-2.5 hours, collecting volatile oil, adding the distilled medicinal residues to an ethanol solution with a volume fraction of 65%-75% at a solid-liquid ratio of 1:8-12 g / mL, extracting it at 60-65° C. for 25-35 minutes, concentrating under reduced pressure, and drying to obtain an ethanol extract, and mixing the volatile oil and the ethanol extract to obtain the tangerine peel extract; more preferably comprises: crushing the tangerine peel, mixing it with water and soaking it for 40 minutes, distilling it at 105° C. for 2 hours, collecting volatile oil, adding the distilled medicinal residues to an ethanol solution with a volume fraction of 70% at a solid-liquid ratio of 1:10 g / mL, extracting it at 63° C. for 30 minutes, concentrating under reduced pressure, and drying to obtain an ethanol extract, and mixing the volatile oil and the ethanol extract to obtain the tangerine peel extract. The tangerine peel extract of the present invention regulates gastrointestinal function by regulating intestinal flora, promotes the absorption of other components, reduces endogenous purine production, and alleviates side effects such as abdominal distension and loose stools that may be caused by diuretics.

[0023] The present invention also provides a method for preparing the above-mentioned theaflavin composition, comprising: mixing hydroxypropyl-β-cyclodextrin with water to obtain a hydroxypropyl-β-cyclodextrin solution, adding theaflavin, chicory extract, Hovenia dulcis fruit extract, Plantago seed extract, Poria cocos extract, Tangerine peel extract, and Corn silk extract, performing ultrasonic inclusion complexation, and spray drying to obtain the theaflavin composition; the concentration of the hydroxypropyl-β-cyclodextrin solution is preferably 15%-20% g / mL by mass volume ratio, more preferably 18% g / mL; the volume ratio of the total weight of theaflavin, chicory extract, Hovenia dulcis fruit extract, Plantago seed extract, Poria cocos extract, Tangerine peel extract, and Corn silk extract to the hydroxypropyl-β-cyclodextrin solution is preferably 1:12-18 g / mL, more preferably 1:15 g / mL. Inclusion of the active ingredients in the theaflavin composition with hydroxypropyl-β-cyclodextrin can improve the solubility and stability of the active ingredients and prevent oxidation of the active ingredients, which would otherwise reduce their efficacy.

[0024] The present invention also provides the use of the above-mentioned theaflavins composition in the preparation of a drug for treating hyperuricemia. The drug comprises the above-mentioned theaflavins composition and pharmaceutically acceptable excipients. The dosage form of the drug includes tablets, capsules, granules, pills, and oral liquid.

[0025] In the present invention, unless otherwise specified, all raw material components are commercially available products well known to those skilled in the art.

[0026] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions of the present invention. Obviously, the embodiments described are only some of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0027] In the following examples, unless otherwise specified, all methods are conventional.

[0028] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0029] Theaflavin was obtained by the preparation method of Example 4 of the invention patent CN117867052A, and the theaflavin content was 63.9% as determined by high performance liquid chromatography; Lactobacillus plantarum grx03 was purchased from the General Microbiology Center of the China Culture Collection Administration of Microorganisms, with a deposit number of CGMCC No. 21268, which is disclosed in CN112795500A; Bifidobacterium animalis subsp. lactis BX-245 was purchased from the General Microbiology Center of the China Culture Collection Administration of Microorganisms, with a deposit number of CGMCC No.26973, disclosed in CN116769654A; cellulase was purchased from Qingdao Haiweisen Biotechnology Co., Ltd., with an enzyme activity of 100,000 U / g, item number M612; xylanase was purchased from Jiangsu Libin Bioengineering Co., Ltd., with an enzyme activity of 10,000 U / g, item number 02144; neutral protease was purchased from Nanjing Xinjiehui Biotechnology Co., Ltd., with an enzyme activity of 50,000 U / g, item number J9; plantain seed extract was purchased from Shanxi Xinyu Biotechnology Co., Ltd., item number 2025010840; corn silk extract was purchased from Shaanxi Xintianyu Biotechnology Co., Ltd., item number XTYYMX.

[0030] The w / v appearing in the Examples and Comparative Examples refers to the mass volume concentration. Taking an 18% (w / v) hydroxypropyl-β-cyclodextrin solution as an example, it means that 18 g of hydroxypropyl-β-cyclodextrin is contained in every 100 mL of the solution.

[0031] Example 1 Theaflavin composition for preventing and treating hyperuricemia (1) Preparation of chicory extract The chicory stems and leaves were crushed, passed through a 200-mesh sieve, mixed with purified water at a solid-liquid ratio of 1:20 g / mL, inoculated with composite microorganisms, fermented at 37°C and pH 6.5 for 66 hours, sterilized, filtered, and the filtrate was concentrated and dried to a water content of 2 wt% to obtain a chicory extract; The composite microorganisms consisted of Lactobacillus plantarum grx03 and Bifidobacterium animalis subspecies lactis BX-245. The bacterial count of Lactobacillus plantarum grx03 in the composite microorganisms was 5×10 9 / g, and the bacterial count of Bifidobacterium animalis subsp. lactis BX-245 was 8×10 8 / g, and the inoculum amount was 1.5% of the mass of chicory stems and leaves; (2) Hovenia dulcis fruit extract The Hovenia dulcis fruit was crushed and passed through a 500-mesh sieve, mixed with a 60% ethanol solution at a solid-liquid ratio of 1:10 g / mL, and ultrasonically extracted twice at a temperature of 45°C and a power of 280 W, each time for 10 min. The filtrates were combined, concentrated, and dried to a water content of 5 wt% to obtain the Hovenia dulcis fruit extract; (3) Poria cocos extract The Poria cocos sclerotia were crushed, passed through an 80-mesh sieve, mixed with water at a solid-liquid ratio of 1:25 g / mL, and a composite enzyme of 1% by weight of the Poria cocos sclerotia was added. The mixture was ultrasonically extracted at a temperature of 52°C, a pH value of 5.2, and a power of 400 W for 20 min, the enzyme was inactivated, and the mixture was filtered. The filtrate was concentrated to 1 / 3 of the volume of the filtrate, and 6 times the volume of a 90% ethanol solution was added for precipitation for 12 h. The precipitate was centrifuged and dried to a water content of 3 wt% to obtain a Poria cocos extract. The complex enzyme consists of cellulase, xylanase and neutral protease in a mass ratio of 6:3:1; (4) Tangerine peel extract The dried tangerine peel was crushed and passed through a 200-mesh sieve, mixed with water at a solid-liquid ratio of 1:8 g / mL, and soaked for 40 minutes. The mixture was then distilled at 105° C. for 2 hours to collect the volatile oil. The distilled medicinal residue was added with a 70% ethanol solution by volume at a solid-liquid ratio of 1:10 g / mL, extracted at 63° C. for 30 minutes, concentrated under reduced pressure, and dried to a water content of 3 wt% to obtain an ethanol extract. The volatile oil and the ethanol extract were mixed to obtain a dried tangerine peel extract. (5) Preparation of theaflavins composition Accurately weigh 18 parts of theaflavins, 18 parts of chicory extract, 10 parts of Hovenia dulcis fruit extract, 12 parts of Plantago seed extract, 12 parts of Poria cocos extract, 7 parts of tangerine peel extract, and 4 parts of corn silk extract; Hydroxypropyl-β-cyclodextrin was mixed with distilled water at 50°C to obtain a hydroxypropyl-β-cyclodextrin solution with a concentration of 18% (w / v). Theaflavin, chicory extract, Hovenia dulcis extract, Plantago seed extract, Poria extract, Tangerine peel extract and Corn silk extract were added at a material-liquid ratio of 1:15 g / mL. The mixture was included at 50°C and 400 rpm for 2.5 hours and spray-dried to obtain a theaflavin composition.

[0032] Example 2 Theaflavin composition for preventing and treating hyperuricemia (1) Preparation of chicory extract The chicory stems and leaves were crushed, passed through a 180-mesh sieve, mixed with purified water at a solid-liquid ratio of 1:18 g / mL, inoculated with composite microorganisms, fermented at 35°C and pH 6 for 72 h, sterilized, filtered, and the filtrate was concentrated and dried to a water content of 3 wt%; The composite microorganisms consisted of Lactobacillus plantarum grx03 and Bifidobacterium animalis subspecies lactis BX-245. The bacterial count of Lactobacillus plantarum grx03 in the composite microorganisms was 4×10 9 / g, and the bacterial count of Bifidobacterium animalis subsp. lactis BX-245 was 5×10 8 / g, the inoculum amount was 1% of the mass of chicory stems and leaves; (2) Hovenia dulcis fruit extract The Hovenia dulcis fruit was crushed through a 400-mesh sieve, mixed with a 55% ethanol solution at a solid-liquid ratio of 1:8 g / mL, and ultrasonically extracted three times at a temperature of 40°C and a power of 250 W, each time for 8 min. The filtrates were combined, concentrated, and dried to a water content of 4 wt% to obtain the Hovenia dulcis fruit extract; (3) Poria cocos extract The Poria sclerotia were crushed, passed through a 50-mesh sieve, mixed with water at a solid-liquid ratio of 1:20 g / mL, and 0.8% of the weight of the Poria sclerotia was added. Ultrasonic extraction was carried out at a temperature of 50°C, a pH value of 5, and a power of 350 W for 25 minutes, the enzyme was inactivated, and the filtrate was concentrated to 1 / 2 of the volume of the filtrate. 4 times the volume of 88% ethanol solution was added for precipitation for 10 hours, and the precipitate was dried to a water content of 4wt% to obtain a Poria extract. The complex enzyme consists of cellulase, xylanase and neutral protease in a mass ratio of 5:2:0.8; (4) Tangerine peel extract The dried tangerine peel was crushed and passed through a 180-mesh sieve, mixed with water at a solid-liquid ratio of 1:6 g / mL, and soaked for 45 minutes. The mixture was then distilled at 102° C. for 2.5 hours to collect the volatile oil. The distilled medicinal residue was added to a 65% ethanol solution by volume at a solid-liquid ratio of 1:8 g / mL, extracted at 60° C. for 35 minutes, concentrated under reduced pressure, and dried to a water content of 5 wt% to obtain an ethanol extract. The volatile oil and the ethanol extract were mixed to obtain a dried tangerine peel extract. (5) Preparation of theaflavins composition Accurately weigh 15 parts of theaflavins, 15 parts of chicory extract, 8 parts of Hovenia dulcis fruit extract, 10 parts of Plantago seed extract, 10 parts of Poria cocos extract, 5 parts of tangerine peel extract, and 3 parts of corn silk extract; Hydroxypropyl-β-cyclodextrin was mixed with distilled water at 48°C to obtain a 15% (w / v) hydroxypropyl-β-cyclodextrin solution, to which theaflavins, chicory extract, Hovenia dulcis fruit extract, Plantago seed extract, Poria cocos extract, Tangerine peel extract, and Corn silk extract were added at a material-liquid ratio of 1:12 g / mL. The mixture was included at 48°C and 380 rpm for 3 h, and spray-dried to obtain a theaflavins composition.

[0033] Example 3 Theaflavin composition for preventing and treating hyperuricemia (1) Preparation of chicory extract The chicory stems and leaves were crushed, passed through a 250-mesh sieve, mixed with purified water at a solid-liquid ratio of 1:23 g / mL, inoculated with composite microorganisms, fermented at 40°C and pH 7 for 60 h, sterilized, filtered, and the filtrate was concentrated and dried to a water content of 4 wt%; The composite microorganisms consisted of Lactobacillus plantarum grx03 and Bifidobacterium animalis subspecies lactis BX-245. The bacterial count of Lactobacillus plantarum grx03 in the composite microorganisms was 7×10 9 / g, and the bacterial count of animal Bifidobacterium lactis subsp. lactis BX-245 was 10×10 8 / g, the inoculation amount is 2% of the weight of chicory stems and leaves; (2) Hovenia dulcis fruit extract The Hovenia dulcis fruit was crushed and passed through a 600-mesh sieve, mixed with a 65% ethanol solution at a solid-liquid ratio of 1:12 g / mL, and ultrasonically extracted at a temperature of 50°C and a power of 300 W for 15 min. The filtrates were combined, concentrated, and dried to a water content of 2 wt% to obtain the Hovenia dulcis fruit extract; (3) Poria cocos extract The Poria sclerotia were crushed, passed through a 100-mesh sieve, mixed with water at a solid-liquid ratio of 1:30 g / mL, and 1.3% of the weight of the Poria sclerotia was added. Ultrasonic extraction was carried out at a temperature of 54°C, a pH value of 5.5, and a power of 450 W for 17 minutes, the enzyme was inactivated, and the filtrate was concentrated to 1 / 4 of the volume of the filtrate. 8 times the volume of 93% ethanol solution was added for precipitation for 10 hours, and the precipitate was dried to a water content of 5wt% to obtain a Poria extract. The complex enzyme consists of cellulase, xylanase and neutral protease in a mass ratio of 7:4:2; (4) Tangerine peel extract The dried tangerine peel was crushed and passed through a 250-mesh sieve, mixed with water at a solid-liquid ratio of 1:10 g / mL, and soaked for 50 minutes. The mixture was then distilled at 107° C. for 1.5 hours to collect the volatile oil. The distilled medicinal residue was added to a 75% ethanol solution by volume at a solid-liquid ratio of 1:12 g / mL, extracted at 65° C. for 35 minutes, concentrated under reduced pressure, and dried to a water content of 5 wt% to obtain an ethanol extract. The volatile oil and the ethanol extract were mixed to obtain a dried tangerine peel extract. (5) Preparation of theaflavins composition Accurately weigh 20 parts of theaflavins, 20 parts of chicory extract, 12 parts of Hovenia dulcis fruit extract, 15 parts of Plantago seed extract, 15 parts of Poria cocos extract, 8 parts of tangerine peel extract, and 5 parts of corn silk extract; Hydroxypropyl-β-cyclodextrin was mixed with distilled water at 53°C to obtain a 20% (w / v) hydroxypropyl-β-cyclodextrin solution. Theaflavin, chicory extract, Hovenia dulcis extract, Plantago seed extract, Poria extract, Tangerine peel extract and Corn silk extract were added at a material-liquid ratio of 1:18 g / mL. The mixture was included at 53°C and 450 rpm for 2 h and spray-dried to obtain a theaflavin composition.

[0034] Comparative Example 1 The specific implementation method is the same as that of Example 1, except that "18 parts of theaflavins and 18 parts of chicory extract" in the preparation of the theaflavins composition in step (5) is replaced with "36 parts of theaflavins", and step (1) is deleted.

[0035] Comparative Example 2 The specific implementation method is the same as that of Example 1, except that "18 parts of theaflavins and 18 parts of chicory extract" in the preparation of the theaflavins composition in step (5) is replaced by "36 parts of chicory extract".

[0036] Comparative Example 3 The specific implementation method is the same as that of Example 1, except that the composite microorganism in step (1) is changed to a single microorganism, Lactobacillus plantarum grx03, with a bacterial count of 5.8×10 9 / g, the inoculation amount was 1.5% of the mass of chicory stems and leaves, the fermentation temperature was 37℃, the pH value was 6.5, and the time was 66h.

[0037] Comparative Example 4 The specific implementation is the same as that of Example 1, except that the "60% by volume ethanol solution" in step (2) is replaced by "pure water".

[0038] Comparative Example 5 The specific implementation method is the same as that of Example 1, except that the preparation method of the Poria cocos extract in step (3) is as follows: crush the Poria cocos sclerotia, pass through an 80-mesh sieve, mix with water at a solid-liquid ratio of 1:25 g / mL, heat and extract at 95°C for 2 h, filter, concentrate the filtrate to 1 / 3 of the volume of the filtrate, add 6 times the volume of 90% ethanol solution, precipitate for 12 h, centrifuge, and dry the precipitate to a water content of 3 wt% to obtain the Poria cocos extract.

[0039] Comparative Example 6 The specific implementation method is the same as that of Example 1, except that the volatile oil in step (4) is the tangerine peel extract.

[0040] Test Example 1 In vitro assay - determination of xanthine oxidase inhibitory activity The xanthine oxidase inhibitory activities of the theaflavin compositions of Examples 1-3 and Comparative Examples 1-6 were respectively tested.

[0041] 10 μg / mL of theaflavin composition and 10 μg / mL of allopurinol (positive control) were prepared in 0.2 M PBS solution at pH 7.5.

[0042] The xanthine oxidase (XOD) activity detection kit (purchased from Beijing Solebow Technology Co., Ltd., model BC1090) was used to detect the xanthine oxidase activity of the tested theaflavins composition, positive control and blank control (not containing the tested component), and the xanthine oxidase inhibition rate was calculated. The specific results are shown in Table 1.

[0043] Xanthine oxidase inhibition rate (%) = (1-absorbance of the test sample / absorbance of the blank control) × 100%.

[0044] Table 1 Inhibition rate of XOD in each group Grouping XOD inhibition rate (%) Grouping XOD inhibition rate (%) Positive control group 74.26±8.13 Comparative Example 2 52.25±4.50 Example 1 72.80±7.48 Comparative Example 3 54.57±4.27 Example 2 69.39±6.31 Comparative Example 4 56.03±5.65 Example 3 70.12±6.84 Comparative Example 5 59.40±6.09 Comparative Example 1 50.64±4.92 Comparative Example 6 62.78±6.76 As shown in Table 1, the examples can significantly improve the xanthine oxidase inhibition rate relative to the comparative examples, which is comparable to the xanthine oxidase inhibition rate of the positive control group, indicating that the theaflavins composition of the present invention can reduce uric acid synthesis and lower blood uric acid levels. As shown in Example 1 and Comparative Examples 1-2, when the dosage is the same, the combination of theaflavins and chicory extract can significantly improve the xanthine oxidase inhibition rate, indicating that theaflavins and chicory extract have a synergistic effect in improving the xanthine oxidase inhibition rate. As shown in the data of Example 1 and Comparative Example 3, Lactobacillus plantarum grx03 and Bifidobacterium animalis subsp. lactis BX-245 as a composite microorganism can better improve the inhibition rate of theaflavins composition on xanthine oxidase than Lactobacillus plantarum grx03 alone. As shown in the data of Example 1 and Comparative Examples 4-6, the theaflavins compositions obtained with different solvents, different extraction methods, and different ingredients have different degrees of influence on the xanthine oxidase inhibition rate. It can be seen that the theaflavins composition of the examples has a high xanthine oxidase inhibition rate, and thus in vivo animal experiments can be conducted to verify its efficacy in treating hyperuricemia.

[0045] Test Example 2 Animal experiments 18-22 g SPF male Kunming mice were selected and adaptively fed for 7 days with free access to food and water. They were then randomly divided into 12 groups (blank group, model group, positive group (allopurinol) and experimental group (Examples 1-3 and Comparative Examples 1-6)), with 10 mice in each group.

[0046] Except for the blank group, mice in the other groups were injected intraperitoneally with 250 mg / kg potassium oxonate and 300 mg / kg hypoxanthine once daily to establish a hyperuricemia mouse model. Two hours after injection, the positive group received 5 mg / kg allopurinol by gavage, while the experimental group received 200 mg / kg by gavage at a volume of 10 mL / kg, once daily for 30 consecutive days.

[0047] After the administration, all mice were fasted but not watered. 24 hours later, the anesthetized mice were sacrificed by cervical dislocation. Blood was collected and centrifuged at 4°C, 3000 rpm for 10 minutes. The upper layer was serum, which was stored for later use. The mouse liver tissue was washed and stored for later use.

[0048] (1) Uric acid and creatinine determination The serum uric acid and creatinine levels of each group were determined using a biochemical analyzer. The specific results are shown in Table 2.

[0049] Table 2 Serum uric acid and creatinine levels in each group Grouping Uric acid (μmol / L) Creatinine (μmol / L) Blank group 157.52±16.43 84.51±6.24 Model Group 324.95±28.20 136.28±15.51 Positive group 211.31±22.75 93.97±9.16 Example 1 197.48±16.91 90.15±7.68 Example 2 203.64±18.07 92.60±9.02 Example 3 199.07±17.49 91.34±8.75 Comparative Example 1 265.49±27.52 112.92±13.59 Comparative Example 2 258.70±25.84 110.79±10.33 Comparative Example 3 246.18±26.16 108.45±12.87 Comparative Example 4 232.23±22.62 105.83±11.94 Comparative Example 5 220.86±19.38 97.06±9.47 Comparative Example 6 218.92±20.42 96.55±10.06 As can be seen from the data in Table 2, the uric acid and creatinine levels in the model group were significantly higher than those in the blank group, indicating that the mouse hyperuricemia model was successfully established. The uric acid and creatinine levels in the serum of the example group and the positive group were not much different, indicating that the theaflavins composition of the example group can effectively inhibit the symptoms of hyperuricemia in the serum. As can be seen from the data comparison of Example 1 and Comparative Examples 1-2, the theaflavins composition lacking theaflavins and chicory extract has no significant effect on reducing uric acid and creatinine, indicating that the combination of theaflavins and chicory extract has a significant effect in treating hyperuricemia. As can be seen from Example 1 and Comparative Examples 3-6, the theaflavins compositions obtained by different microbial fermentations, different solvents, different extraction methods, and different ingredients have different effects on reducing the levels of uric acid and creatinine. This indicates that the theaflavins composition of the present invention has the effect of treating hyperuricemia.

[0050] (2) Determination of xanthine oxidase activity The activity of xanthine oxidase in serum and liver tissue of each group was determined according to the kit instructions (purchased from Beijing Solebow Technology Co., Ltd., model BC1090). The specific results are shown in Table 3.

[0051] Table 3 Xanthine oxidase activities in serum and liver of each group Grouping Serum XOD activity (U / g mass) Liver XOD activity (U / mL) Blank group 9.42±0.87 14.32±0.91 Model Group 25.69±1.63 36.90±1.80 Positive group 12.56±1.01 15.26±1.36 Example 1 13.30±0.82 16.43±1.23 Example 2 14.08±1.16 17.18±1.48 Example 3 13.71±0.90 16.51±1.15 Comparative Example 1 18.94±1.48 22.88±1.60 Comparative Example 2 18.53±1.64 23.04±1.79 Comparative Example 3 18.27±1.21 20.69±1.52 Comparative Example 4 17.84±1.59 19.17±1.54 Comparative Example 5 17.15±1.35 19.65±1.37 Comparative Example 6 16.47±1.28 18.97±1.18 As shown in Table 3, the positive group and the example group significantly reduced xanthine oxidase activity, with comparable efficacy. The example group reduced xanthine oxidase activity more significantly than the comparative example group. The data from Example 1 and Comparative Examples 1-6 indicate that the omission of active ingredients, theaflavin compositions obtained using different microbial fermentations, different solvents, different extraction methods, and different ingredients can affect xanthine oxidase activity in serum and liver. This suggests that the theaflavin extracts of the present invention can achieve the therapeutic effect of hyperuricemia by reducing xanthine oxidase activity.

[0052] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A theaflavin composition for preventing and treating hyperuricemia, characterized in that: The invention comprises the following raw materials in parts by weight: 15-20 parts of theaflavins, 15-20 parts of chicory extract, 8-12 parts of hovenia dulcis fruit extract, 10-15 parts of plantain seed extract, 10-15 parts of poria extract, 5-8 parts of tangerine peel extract and 3-5 parts of corn silk extract.

2. The theaflavin composition according to claim 1, characterized in that The chicory extract is obtained by liquid anaerobic fermentation of composite microorganisms.

3. The theaflavin composition according to claim 2, characterized in that The composite microorganism consists of Lactobacillus plantarum grx03 and Bifidobacterium animalis subspecies lactis BX-245.

4. The theaflavin composition according to claim 1, characterized in that The Hovenia dulcis fruit extract is obtained by ultrasonic extraction with ethanol.

5. The theaflavin composition according to claim 1, characterized in that The Poria cocos extract is obtained by complex enzyme hydrolysis and ethanol precipitation.

6. The theaflavin composition according to claim 5, characterized in that The complex enzyme consists of cellulase, xylanase and neutral protease.

7. The theaflavin composition according to claim 1, characterized in that The tangerine peel extract is subjected to distillation to obtain volatile oil, the distilled medicinal residue is subjected to ethanol extraction to obtain an ethanol extract, and the volatile oil and the ethanol extract are mixed to obtain the tangerine peel extract.

8. A method for preparing the theaflavin composition according to any one of claims 1 to 7, characterized in that: include: Hydroxypropyl-β-cyclodextrin is mixed with water to obtain a hydroxypropyl-β-cyclodextrin solution, and theaflavin, chicory extract, Hovenia dulcis fruit extract, Plantago seed extract, Poria cocos extract, Tangerine peel extract and Corn silk extract are added thereto, followed by ultrasonic inclusion and spray drying to obtain the theaflavin composition.

9. The preparation method according to claim 8, characterized in that The volume ratio of the total weight of theaflavins, chicory extract, Hovenia dulcis fruit extract, Plantago seed extract, Poria cocos extract, Tangerine peel extract and Corn silk extract to the hydroxypropyl-β-cyclodextrin solution is 1:12-18 g / mL.

10. Use of the theaflavin composition according to any one of claims 1 to 7 in the preparation of a drug for treating hyperuricemia.