Composition for reducing uric acid and preparation method thereof

Abstract

The invention discloses a composition for reducing uric acid. The composition is prepared from the following raw materials in parts by weight: 10 to 30 parts of fructus phyllanthi, 15 to 20 parts of fructus gardeniae, 15 to 20 parts of radix astragali seu hedysari and 35 to 50 parts of herba cichorii. The invention also provides a preparation method of the composition for reducing uric acid. The composition prepared by the invention is simple in component, high in XOD and URAT1 dual inhibitory activity, antioxidant activity and antiglycemic activity, and high in bioavailability of active components.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine fermentation, and particularly relates to a composition for reducing uric acid and a preparation method thereof. Background Art

[0002] Uric acid is the final product of purine metabolism in the human body and is mainly excreted by the kidneys. When uric acid is produced excessively or its excretion is impaired, the uric acid concentration in the blood will increase, leading to hyperuricemia, gout, etc. In recent years, with the changes in people's dietary structure (such as increased intake of high-protein and high-purine foods), the prevalence of hyperuricemia and gout has increased significantly. According to the "Dietary Guidelines for Hyperuricemia and Gout in Adults (2024 Edition)", the prevalence of hyperuricemia among adult residents in my country is 14%, and the prevalence of gout is 0.86% to 2.20%, which has become a major problem threatening public health.

[0003] Currently, lowering uric acid levels (including inhibiting uric acid production and promoting uric acid excretion) remains a core strategy for treating hyperuricemia. Xanthine oxidase (XOD) is a key enzyme in purine metabolism, and abnormal activity can lead to increased uric acid production. The uric acid transporter (URAT1), as the primary pathway for uric acid reabsorption, is an important target for uricosuric drugs. The inhibition rates of XOD and URAT1 can be used to directly reflect the reduction of uric acid levels.

[0004] Traditional Chinese medicine (TCM), with its advantages of multi-target regulation and significant efficacy, has gradually become a research hotspot for the treatment of hyperuricemia. However, existing TCM compositions still face the following challenges: First, their complex composition leads to cumbersome preparation processes and unclear uric acid-lowering effects. Second, traditional extraction methods (such as water extraction) have low dissolution rates, resulting in insufficient bioavailability of active ingredients and difficulty in fully exerting their efficacy.

[0005] Therefore, there is an urgent need to develop a uric acid-lowering composition that has both efficient XOD and URAT1 dual inhibitory activity, streamlined ingredients, and high bioavailability. Summary of the Invention

[0006] In order to solve the above technical problems, a uric acid-lowering composition is developed that can enhance the dual inhibitory activity of XOD and URAT1, has streamlined ingredients and high bioavailability. The present application provides a uric acid-lowering composition and a preparation method thereof.

[0007] In a first aspect, the present application provides a composition for lowering uric acid, comprising the following raw materials in parts by weight: 10 to 30 parts of emblica fruit, 15 to 20 parts of gardenia fruit, 15 to 25 parts of astragalus root, and 35 to 50 parts of chicory.

[0008] By adopting the above technical solution, the present application has simple components, realizes the regulation of the "generation-excretion" dual pathway of uric acid metabolism, and has high bioavailability of active ingredients. Among them, Phyllanthus emblica has the effects of lowering blood uric acid levels, inhibiting gouty inflammation, and analgesic effects; Gardenia jasminoides can inhibit XOD activity and regulate the expression of uric acid transporters; Astragalus membranaceus can relieve acute hyperuricemia by reducing uric acid production and kidney protection, and promote uric acid excretion by regulating uric acid transporters; Chicory can reduce uric acid by reducing uric acid metabolic enzymes and key reaction enzymes, such as reducing the activity of xanthine oxidase, thereby achieving uric acid reduction. It can also start from the kidneys and intestinal excretion, inhibit reabsorption and regulate the intestinal environment to achieve a balance in uric acid metabolism. The synergistic combination of the four drugs significantly improves the dual inhibitory activity of XOD and URAT1, thereby reducing uric acid.

[0009] Optionally, the composition comprises the following raw materials in parts by weight: 10 parts of emblica fruit, 20 parts of gardenia fruit, 20 parts of chicory, and 50 parts of astragalus.

[0010] Through the above technical scheme, the present application adopts a specific ratio of emblica, gardenia, astragalus, and chicory, which can improve the dual inhibitory activity of XOD and URAT1 while also increasing the active ingredients such as total saponins, crude polysaccharides, total sugars, total flavonoids, as well as the antioxidant activity and anti-glycemic activity.

[0011] Optionally, the raw materials of the composition further include one of tuna peptide powder and sea cucumber peptide.

[0012] Optionally, the raw materials of the composition further include 2 parts by weight of tuna peptide powder and 2 parts by weight of sea cucumber peptide.

[0013] Through the above technical solution, the present application also contains tuna peptide powder and sea cucumber peptide to participate in lowering uric acid. Tuna peptide powder can effectively scavenge hydroxyl radicals and DPPH free radicals, thereby inhibiting the production of uric acid caused by oxidative stress. Sea cucumber peptide can bind to the molybdenum pterin cofactor in the XOD active center through hydrogen bonds and hydrophobic interactions, inhibiting its ability to catalyze the conversion of xanthine to uric acid. The combination of the two can improve the dual inhibitory activity of XOD and URAT1, total saponins, crude polysaccharides, total sugars, total flavonoids and other active ingredients as well as antioxidant activity and anti-glycemic activity.

[0014] Optionally, the composition is obtained by fermenting the raw materials with Lactobacillus plantarum.

[0015] By adopting the above technical solution, the present application adopts inoculation of Lactobacillus plantarum to improve the bioavailability of active ingredients such as total saponins, crude polysaccharides, total sugars, and total flavonoids.

[0016] In a second aspect, the present invention provides a method for preparing a composition for reducing uric acid, comprising the following steps: S1. Pretreatment: weigh the raw materials according to the ratio, crush them to obtain Chinese medicine powder; S2, fermentation: the Chinese medicine powder and sterile water are mixed evenly to obtain a mixture, and then Lactobacillus plantarum is added, and the mixture is fermented at 35-40°C for 12-72 hours, and then the temperature is lowered to 26-28°C to continue fermentation; S3. Purification: filtration, sterilization, evaporation and concentration.

[0017] Optionally, in S1, the crushing mesh size is 60 to 80 meshes.

[0018] Optionally, in S2, the weight ratio of sterile water to traditional Chinese medicine powder is (9-11):1, and the volume of Lactobacillus plantarum is 3%-5% of the total volume of the mixed solution.

[0019] Optionally, in S2, at least one of tuna peptide powder and sea cucumber peptide is further added to the mixed solution, the ratio of the first fermentation time to the continued fermentation time is (0.5-2):1, and the fermentation is carried out in a shaking table.

[0020] Optionally, in S3, the sterilization temperature is 105-120° C., the evaporation temperature is 50-60° C., and the mixture is concentrated to 20-30% of the volume of the mixed solution.

[0021] When the raw materials of the composition further include tuna peptide powder and / or sea cucumber peptide, the preparation method of the composition for reducing uric acid comprises the following steps: S1. Pretreatment: weigh the raw materials according to the ratio, crush them to obtain Chinese medicine powder, add tuna peptide powder and / or sea cucumber peptide to obtain a mixture powder; S2, fermentation: the mixture powder is mixed with sterile water to obtain a mixture, and then Lactobacillus plantarum is added, and the mixture is fermented at 35-40°C for 12-72 hours, and then cooled to 26-28°C to continue fermentation; S3. Purification: filtration, sterilization, evaporation and concentration.

[0022] Optionally, in S2, the weight ratio of sterile water to the mixture powder is (9-11):1.

[0023] By adopting the above technical solution, the present application adopts a two-stage temperature fermentation, which can avoid the degradation of heat-sensitive ingredients, improve the bioavailability of active ingredients, and thereby improve the dual inhibitory activity of XOD and URAT1, antioxidant activity and anti-glycemic activity.

[0024] In summary, this application includes at least one of the following beneficial technical effects: 1. The present invention has simple ingredients, achieves dual regulation of the "production-excretion" pathway of uric acid metabolism, and has high bioavailability of the active ingredients; 1. This application uses a specific ratio of emblica, gardenia, astragalus, and chicory to improve the dual inhibitory activity of XOD and URAT1 while also increasing the active ingredients such as total saponins, crude polysaccharides, total sugars, and total flavonoids, as well as the antioxidant activity and anti-glycemic activity; 3. This application uses inoculation with Lactobacillus plantarum to improve the bioavailability of active ingredients such as total saponins, crude polysaccharides, total sugars, and total flavonoids; 4. The present application adopts a two-stage temperature fermentation, which can avoid the degradation of heat-sensitive ingredients, improve the bioavailability of active ingredients, and thereby enhance the dual inhibitory activity of XOD and URAT1, antioxidant activity, and anti-glycemic activity. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 Schematic diagram of the effect of the composition of the present application on URAT1 expression. A: URAT1 and β-actin protein bands; B: Relative URAT1 expression. 1# shows the in vitro URAT1 expression of Comparative Example 1, and 2# shows the in vitro URAT1 expression of Example 1. ### p < 0.001 vs. Control, ***p < 0.001 vs. Model. DETAILED DESCRIPTION

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

[0027] The present application designs a composition for lowering uric acid, which comprises the following raw materials in parts by weight: 10 to 30 parts of emblica fruit, 15 to 20 parts of gardenia fruit, 15 to 25 parts of astragalus root, and 35 to 50 parts of chicory.

[0028] The uric acid-lowering composition of the present application is prepared by the following method, comprising the following steps: S1. Pretreatment: Weigh the Chinese medicine raw materials according to the ratio, crush them into 60-80 mesh, and obtain Chinese medicine powder; S2, fermentation: Chinese medicine powder and sterile water are mixed, the weight ratio of sterile water and Chinese medicine powder is (9~11): 1, obtains mixed solution, then adds the plant lactobacillus culture fluid of 3%~5% of mixed solution cumulative volume, is placed in shaking table, after 35~40 ℃ of first fermentations 12~72h, is cooled to 26~28 ℃ and continues to ferment 24~60h; S3. Purification: Filter, sterilize at 105-120°C, and evaporate and concentrate at 50-60°C to 20-30% of the volume of the mixed solution.

[0029] The raw materials used in the examples of this application can be obtained from commercial sources, including: Phyllanthus emblica, Anhui Mudantianxia Technology Co., Ltd.; Gardenia, Anhui Mudantianxia Technology Co., Ltd.; Astragalus, Anhui Mudantianxia Technology Co., Ltd.; Chicory, Anhui Mudantianxia Technology Co., Ltd.; Lactobacillus plantarum, Hebei Yiran Biotechnology Co., Ltd.; Tuna peptide powder, Yantai Jiahui Marine Biotechnology Co., Ltd.; Sea cucumber peptide, Yantai Jiahui Marine Biotechnology Co., Ltd.

[0030] Test items and methods 1. Determination of XOD inhibitory activity and half inhibitory concentration (IC50) value in vitro: Prepare a 0.5 U / mL xanthine oxidase solution and a 2.0 μmol / L xanthine solution in 0.2 mol / L phosphate buffer. Add 50 μL of the xanthine oxidase solution to 200 μL of the test solution and mix. Incubate at 25°C for 15 minutes, then add 400 μL of the xanthine solution. Continue incubation for 20 minutes, then terminate the reaction by adding 50 μL of a 1 mol / L hydrochloric acid solution. Measure the absorbance (A3) at 290 nm to calculate the XOD activity inhibition rate. Also, use the absorbance (A0) measured without the test solution and xanthine oxidase solution, the absorbance (A1) measured without the test solution, and the absorbance (A2) measured without the xanthine oxidase solution. XOD activity inhibition rate = [(A1 - A0) - (A3 - A2)] / (A1 - A0) * 100%. The test solution was diluted 2, 4, 8, and 16 times with 55% ethanol, and the absorbance was measured according to the above method. The dose-effect curve was drawn to obtain the IC50 value.

[0031] 2. In vitro URAT1 inhibitory activity assay: Human renal tubular epithelial cells (HK-2) were cultured at a rate of 7.2×10 5 Cells were plated in 6 cm dishes at 400 μg / well and treated 24 hours after plating. MEM cell culture medium served as the control group, 100 mg / L uric acid as the model group, 100 mg / L uric acid combined with 10 μM benzbromarone as the positive control group, and 100 mg / L uric acid combined with 250 mg / L test solution as the experimental group. Protein was extracted from each treatment group after 48 hours of culture, and the effect of each treatment group on URAT1 expression was analyzed by Western blotting using β-actin as an internal reference. The experiment was repeated three times. URAT1 activity inhibition rate = (URAT1 expression level in model group - URAT1 expression level in experimental group) / URAT1 expression level in model group * 100%.

[0032] 3. Determination of active ingredients (total saponins, crude polysaccharides, total sugars, total flavonoids) content 3.1. Determination of total saponin content: The total saponin content of the test solution was determined using the vanillin-glacial acetic acid colorimetric method. Oleanolic acid was used as the standard, and a standard curve was drawn. The total saponin content of the test solution was determined at 543 nm. 3.2. Determination of crude polysaccharide content: Determine the crude polysaccharide content of the test solution in accordance with the entry-exit inspection and quarantine industry standard SN / T 4260-2015 “Determination of crude polysaccharides in plant-derived foods for export – Phenol-sulfuric acid method”; 3.3. Determination of total sugar content: The total sugar content of the test solution was determined using the phenol-sulfuric acid colorimetric method. Glucose was used as the standard, and a standard curve was drawn. The total sugar content of the test solution was measured at 490 nm. 3.4 Determination of Total Flavonoids: The total flavonoids content of the test solution was determined using the aluminum nitrate-sodium hydroxide colorimetric method. Rutin was used as the standard, and a standard curve was drawn. The total flavonoids content of the test solution was determined at 510 nm.

[0033] 4. Determination of antioxidant activity 4.1. Hydroxyl Radical Scavenging Rate: Mix 50 μL of the test solution, 50 μL of a 2 mmol / L ferrous sulfate solution, and 50 μL of a 6 mmol / L hydrogen peroxide solution. Let the mixture stand for 10 minutes. Add 50 μL of a salicylic acid solution and heat at 37°C for 15 minutes. Measure the absorbance of the solution at 510 nm (A1). The absorbance measured without the addition of the test solution is designated as A0, and the absorbance measured without the addition of hydrogen peroxide is designated as A2. Repeat the experiment three times. Hydroxyl Radical Scavenging Rate = [(A1 - A2) / A0] * 100%; 4.2. DPPH Scavenging Rate: Mix 50 μL of the test solution and 150 μL of the DPPH solution. Heat at 37°C for 30 minutes, and measure the absorbance of the solution at 517 nm (A1). Replace the test solution with an equal volume of 70% ethanol, and measure the absorbance as A2. The absorbance measured without adding the test solution is A0. Repeat the experiment three times. DPPH Scavenging Rate = [1-(A1-A2) / A0] * 100%. 4.3. ABTS Scavenging Rate: Mix 50 μL of the test solution and 150 μL of the ABTS solution, and allow to react at room temperature for 30 minutes. Measure the absorbance of the solution at 735 nm (A1). Replace the test solution with an equal volume of 70% ethanol, and measure the absorbance as A2. The absorbance measured without adding the test solution is A0. Repeat the experiment three times. ABTS Scavenging Rate = [1-(A1-A2) / A0] * 100%.

[0034] 5. Anti-glycemic activity determination The Lineweaver-Burk double reciprocal method was used to determine the inhibition rate of the test solution on α-glucosidase activity. Specific embodiments

[0035] Preparation Example 1 Preparation of Lactobacillus plantarum culture solution: Live Lactobacillus plantarum was cultured anaerobically at 37°C for 48 hours; gradient dilution was performed and plated to obtain single colonies; a single colony was picked and cultured for further 12 hours, and then inoculated into MRS liquid culture medium at a 2% inoculum volume and cultured at 37°C for 24 hours.

[0036] Example 1 1 g of emblica fructus, 2 g of gardenia jasminoides, 2 g of astragalus membranaceus, and 5 g of chicory were weighed and crushed to 60 mesh to obtain a Chinese medicine powder. The Chinese medicine powder was evenly mixed with sterile water at a weight ratio of sterile water to Chinese medicine powder of 10:1 to obtain a mixed solution. 3% of the total volume of the mixed solution was then added to the Lactobacillus plantarum culture solution obtained in Preparation Example 1. The mixture was placed in a shaker and fermented at 37° C. for 12 hours, then cooled to 28° C. and fermented for another 60 hours. The mixture was filtered, sterilized at 120° C., and concentrated by evaporation at 60° C. to 20% of the volume of the mixed solution, thereby obtaining a fermented broth of a composition for reducing uric acid.

[0037] Example 2 2 g of emblica fructus, 1.5 g of gardenia jasminoides, 2.5 g of astragalus root, and 4 g of chicory were weighed and crushed to 80 mesh to obtain a Chinese medicine powder. The Chinese medicine powder was evenly mixed with sterile water at a weight ratio of sterile water to Chinese medicine powder of 9:1 to obtain a mixed solution. 5% of the total volume of the mixed solution was then added to the Lactobacillus plantarum culture solution obtained in Preparation Example 1. The mixture was placed in a shaker and fermented at 40° C. for 12 hours, then cooled to 27° C. and fermented for another 60 hours. The mixture was filtered, sterilized at 110° C., and evaporated and concentrated at 55° C. to 25% of the volume of the mixed solution, thereby obtaining a fermented solution of a composition for reducing uric acid.

[0038] Example 3 3 g of emblica fructus, 2 g of gardenia jasminoides, 1.5 g of astragalus root, and 3.5 g of chicory were weighed and crushed to 80 mesh to obtain a Chinese medicine powder. The Chinese medicine powder was evenly mixed with sterile water at a weight ratio of sterile water to Chinese medicine powder of 11:1 to obtain a mixed solution. 5% of the total volume of the mixed solution was then added to the Lactobacillus plantarum culture solution obtained in Preparation Example 1. The mixture was placed in a shaker and fermented at 35° C. for 12 hours, then cooled to 26° C. and fermented for another 60 hours. The mixture was filtered, sterilized at 105° C., and concentrated by evaporation at 50° C. to 30% of the volume of the mixed solution, thereby obtaining a fermented broth of a composition for reducing uric acid.

[0039] The fermentation broth of the uric acid-lowering composition obtained in Examples 1-3 was subjected to in vitro XOD inhibitory activity, IC50 value, in vitro URAT1 inhibitory activity, active ingredients (total saponins, crude polysaccharides, total sugars, total flavonoids), antioxidant activity, and anti-glycemic activity assays. The test results are shown in Table 1: Table 1 Uric acid reduction and biological activity determination of the fermentation broth of the compositions of Examples 1-3 As can be seen from Examples 1-3 and Table 1, the fermentation broth of the composition for reducing uric acid obtained in the present application is obtained by fermenting a specific ratio of emblica, gardenia, astragalus, and chicory. The pH value decreases after fermentation, indicating that the fermentation system is successful. The present application uses a specific ratio of emblica, gardenia, astragalus, and chicory to improve the dual inhibitory activity of XOD and URAT1 while also increasing the active ingredients such as total saponins, crude polysaccharides, total sugars, and total flavonoids, as well as the antioxidant activity and anti-glycemic activity.

[0040] Comparative Example 1 The difference between this comparative example and Example 1 is that in this comparative example, the Chinese medicinal raw materials in Example 1 (1 g of emblica buds, 2 g of gardenia jasminoides, 2 g of astragalus, and 5 g of chicory) are added to sterile water at a material-liquid ratio of 1:10, heated in a water bath, boiled and extracted for 1.5 h, and the filtrate is collected after filtration; the above steps are repeated 3 times, the combined filtrates are evaporated and concentrated to obtain an extract of the same volume as the fermentation broth of the uric acid-lowering composition in Example 1, and in this comparative example, an equal volume of the fermentation broth of the composition is replaced with the extract.

[0041] Comparative Example 2 The difference between this comparative example and Example 1 is that this comparative example is obtained by fermentation at 28° C. for 72 hours.

[0042] Comparative Example 3 The difference between this comparative example and Example 1 is that, in this comparative example, the mass of emblica buds is replaced by gardenia jasminoides.

[0043] Comparative Example 4 The difference between this comparative example and Example 1 is that chicory is not added in this comparative example.

[0044] Comparative Example 5 The difference between this comparative example and Example 1 is that, in this comparative example, Gardenia jasminoides is replaced by Emblica officinalis.

[0045] Comparative Example 6 The difference between this comparative example and Example 1 is that no astragalus is added in this comparative example.

[0046] The fermentation broths of the uric acid-lowering compositions obtained in Example 1 and Comparative Examples 1-5 were assayed for in vitro XOD inhibitory activity, IC50 values, in vitro URAT1 inhibitory activity, active ingredients (total saponins, crude polysaccharides, total sugars, total flavonoids), antioxidant activity, and anti-glycemic activity. The test results are shown in Table 2: Table 2 Uric acid reduction and biological activity determination of fermentation broth of Example 1 and Comparative Examples 1-5 It can be seen from Example 1, Comparative Example 1 and Table 2 that compared with the extract, the IC50 value of the composition is reduced, and the XOD activity inhibition rate, URAT1 activity inhibition rate, active ingredient (total saponins, crude polysaccharides, total sugars, total flavonoids) content, antioxidant activity, and anti-glycemic activity are increased. This is mainly because Lactobacillus plantarum fermentation can produce organic acids, resulting in reduced acidity, and by decomposing plant cell walls, releasing active ingredients such as saponins, polysaccharides, and flavonoids, thereby improving bioavailability and thereby improving the dual inhibitory activity of XOD and URAT1, antioxidant activity, and anti-glycemic activity; the present application adopts a fermentation treatment of the traditional Chinese medicine composition, which can significantly improve the utilization rate of the biologically active ingredients, thereby improving the dual inhibitory activity of XOD and URAT1, antioxidant activity, and anti-glycemic activity.

[0047] It can be seen from Example 1, Comparative Example 2 and Table 2 that when fermentation is continued at 28°C, the bacterial growth is slow and the metabolic activity is insufficient, resulting in reduced dissolution of primary metabolites (such as crude polysaccharides and total sugars) and reduced enzymatic conversion efficiency of secondary metabolites (such as saponins and flavonoids); the present application adopts a two-stage temperature fermentation, the high temperature stage of 37°C promotes the rapid proliferation of plant lactobacillus, accelerates cell wall decomposition, and releases primary metabolites such as polysaccharides and saponins, and the low temperature stage of 28°C avoids the degradation of heat-sensitive components (flavonoids, saponins, etc.). Maintaining two-stage temperature fermentation can balance metabolic efficiency and component stability, thereby significantly improving the dual inhibitory effect of XOD and URAT1, and improving antioxidant activity and anti-glycemic activity.

[0048] As shown in Example 1, Comparative Example 3, and Table 2, when emblica buds were replaced with gardenia jasmine, although gardenia jasmine itself had the effect of inhibiting XOD, it destroyed the "production-excretion" dual-target synergistic mechanism of the original formula (emblica buds were responsible for excretion regulation, and gardenia jasmine focused on production inhibition), resulting in a significant decrease in the URAT1 inhibition rate. In addition, the absence of emblica buds directly reduced active ingredients such as saponins and flavonoids, weakened the antioxidant and anti-glycemic activities, and slightly decreased the crude polysaccharide and total sugar contents.

[0049] It can be seen from Example 1, Comparative Example 4 and Table 2 that without the addition of chicory, the fermentation broth of the obtained composition not only significantly reduced the inhibition rate of XOD and URAT1, but also significantly reduced the content of active ingredients such as crude polysaccharides and total sugars, and weakened the antioxidant and anti-glycemic activities.

[0050] As shown in Example 1, Comparative Example 5 and Table 2, after Gardenia was replaced by Emblica officinalis, the composition fermentation liquid lost its direct inhibitory effect on XOD (Gardenia officinalis is the core target), resulting in a significant decrease in the XOD activity inhibition rate. Although Emblica officinalis can partially compensate for the URAT1 inhibitory effect through the excretion pathway, the overall synergistic mechanism is broken, and the URAT1 inhibition rate is still slightly reduced. In addition, total flavonoids and antioxidant activity are significantly reduced due to the absence of Gardenia officinalis.

[0051] It can be seen from Example 1, Comparative Example 6 and Table 2 that without the addition of Astragalus, the XOD activity inhibition rate decreased slightly, but the URAT1 activity inhibition rate decreased significantly, and the total saponin content was significantly reduced, and the antioxidant and anti-glycemic activities were weakened.

[0052] It can be seen from Example 1, Comparative Examples 3-6 and Table 2 that the present application adopts the synergistic combination of emblica, gardenia, astragalus and chicory to achieve dual high-efficiency inhibition of XOD and URAT1, while taking into account highly active ingredients (total saponins, polysaccharides, flavonoids) and comprehensive biological activities (antioxidant, anti-glycemic).

[0053] Example 4 1 g of emblica fructus, 2 g of gardenia jasminoides, 2 g of astragalus membranaceus, and 5 g of chicory were weighed and crushed to 60 mesh to obtain a traditional Chinese medicine powder. 0.4 g of tuna peptide powder was added to obtain a mixture powder. The mixture powder was evenly mixed with sterile water at a weight ratio of sterile water to mixture powder of 10:1 to obtain a mixed solution. 3% of the total volume of the mixed solution was then added to the Lactobacillus plantarum culture solution obtained in Preparation Example 1. The mixture was placed in a shaker and fermented at 37°C for 12 hours. The mixture was then cooled to 28°C and fermented for another 60 hours. The mixture was filtered, sterilized at 120°C, and concentrated by evaporation at 60°C to 20-30% of the volume of the mixed solution, thereby obtaining a fermented solution of a composition for reducing uric acid.

[0054] Example 5 The difference between this embodiment and embodiment 4 is that in this embodiment, the tuna peptide powder is replaced with sea cucumber peptide.

[0055] Example 6 The difference between this embodiment and embodiment 4 is that in this embodiment, 0.4 g of tuna peptide powder is replaced with 0.2 g of tuna peptide powder and 0.2 g of sea cucumber peptide.

[0056] The fermentation broths of the uric acid-lowering compositions obtained in Examples 1 and 4-6 were assayed for in vitro XOD inhibitory activity, IC50 values, in vitro URAT1 inhibitory activity, active ingredients (total saponins, crude polysaccharides, total sugars, total flavonoids), antioxidant activity, and anti-glycemic activity. The test results are shown in Table 3: Table 3 Uric acid reduction and biological activity determination of the fermentation broth of the compositions of Example 1 and Examples 4-6 It can be seen from Example 1, Examples 4-6 and Table 3 that the addition of tuna peptide powder alone can significantly improve the URAT1 inhibition rate and DPPH clearance rate, and the addition of sea cucumber peptide alone can significantly improve the XOD activity inhibition rate and IC50 value. The combined use of tuna peptide powder and sea cucumber peptide can improve the dual inhibitory activity of XOD and URAT1, total saponins, crude polysaccharides, total sugars, total flavonoids and other active ingredients as well as antioxidant activity and anti-glycemic activity.

[0057] Example 7 The difference between this embodiment and embodiment 6 is that this embodiment adopts fermentation at 37° C. for 24 hours, cooling to 28° C. and continuing fermentation for 48 hours.

[0058] Example 8 The difference between this embodiment and embodiment 6 is that this embodiment adopts fermentation at 37° C. for 48 hours, cooling to 28° C. and continuing fermentation for 24 hours.

[0059] Example 9 The difference between this embodiment and embodiment 6 is that this embodiment is obtained by fermentation at 37° C. for 72 hours.

[0060] The fermentation broths of the uric acid-lowering compositions obtained in Examples 6 and 7-9 were assayed for in vitro XOD inhibitory activity, IC50 values, in vitro URAT1 inhibitory activity, active ingredients (total saponins, crude polysaccharides, total sugars, total flavonoids), antioxidant activity, and anti-glycemic activity. The test results are shown in Table 4. Table 4 Uric acid reduction and biological activity determination of the fermentation broth of the compositions of Example 6 and Examples 7-9 As shown in Example 5, Examples 7-9 and Table 4, the present application adopts a two-stage temperature fermentation. The first 24h high temperature fermentation (37°C) stage can promote the rapid growth of microorganisms and the accumulation of primary metabolites (such as polysaccharides and saponins); the latter 48h low temperature (28°C) stage can slow down the growth rate of microorganisms, prolong the synthesis cycle of secondary metabolites (such as saponins and flavonoids), and avoid the degradation of polypeptides and flavonoids. Excessive high temperature fermentation (37°C) can lead to metabolic imbalance, degradation or consumption of active ingredients, and decomposition of crude polysaccharides into monosaccharides, while the other indicators fall back. The present application adopts a specific temperature and fermentation time for fermentation, which can significantly increase the active ingredients such as total saponins, crude polysaccharides, total sugars, and total flavonoids without reducing the inhibitory activity of XOD and URAT1.

[0061] The embodiments of this specific implementation method are all preferred embodiments of the present application and are not intended to limit the scope of protection of the present application. Therefore, any simple modifications, equivalent replacements and improvements based on the technical solutions of the present application should be covered within the scope of protection of the present application.

Claims

1. A composition for reducing uric acid, characterized in that: The invention comprises the following raw materials in parts by weight: 10-30 parts of emblica fruit, 15-20 parts of gardenia fruit, 15-25 parts of astragalus root and 35-50 parts of chicory.

2. A uric acid-lowering composition according to claim 1, characterized in that: The composition comprises the following raw materials in parts by weight: 10 parts of emblica fruit, 20 parts of gardenia fruit, 20 parts of astragalus root and 50 parts of chicory.

3. A composition for reducing uric acid according to claim 1, characterized in that The raw materials of the composition also include one of tuna peptide powder and sea cucumber peptide.

4. A composition for reducing uric acid according to claim 3, characterized in that: The raw materials of the composition further include 2 parts by weight of tuna peptide powder and 2 parts by weight of sea cucumber peptide.

5. A uric acid-lowering composition according to claim 1, characterized in that: The composition is obtained by fermenting the raw materials through Lactobacillus plantarum.

6. A method for preparing the uric acid-lowering composition according to claim 1, characterized in that: The steps include: S1. Pretreatment: weigh the raw materials according to the ratio, crush them to obtain Chinese medicine powder; S2, fermentation: the Chinese medicine powder and sterile water were mixed evenly to obtain a mixture, and then Lactobacillus plantarum culture solution was added. After the first fermentation at 35-40°C for 12-72 hours, the temperature was lowered to 26-28°C and the fermentation was continued; S3. Purification: filtration, sterilization, evaporation and concentration.

7. The method for preparing the uric acid-lowering composition according to claim 6, wherein: In the S1, the crushing mesh size is 60-80 meshes.

8. The method for preparing the uric acid-lowering composition according to claim 6, wherein: In the S2, the weight ratio of sterile water to traditional Chinese medicine powder is (9-11):1, and the volume of Lactobacillus plantarum culture solution is 3%-5% of the total volume of the mixed solution.

9. The method for preparing the uric acid-lowering composition according to claim 6, wherein: In S2, the ratio of the first fermentation time to the continued fermentation time is (0.5-2):1, and the fermentation is carried out in a shaking table.

10. The method for preparing the uric acid-lowering composition according to claim 6, wherein: In the S3, the sterilization temperature is 105-120° C., the evaporation temperature is 50-60° C., and the mixture is concentrated to 20-30% of the volume of the mixed solution.

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