A composition for reducing uric acid and a method for preparing the same

Abstract

The application discloses a composition for reducing uric acid, which comprises the following raw materials in parts by weight: 10-30 parts of Phyllanthus emblica, 15-20 parts of Gardenia, 15-20 parts of Astragalus, and 35-50 parts of Chichorium intybus. The application also provides a preparation method of the composition for reducing uric acid. The composition prepared by the application has simple components, high XOD and URAT1 double inhibition activity, high antioxidant activity and high anti-hyperglycemic activity, and high bioavailability of active ingredients.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine fermentation technology, and specifically relates to a composition for lowering uric acid and its preparation method. Background Technology

[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 in excess or excreted improperly, the concentration of uric acid in the blood increases, leading to hyperuricemia, gout, and other conditions. In recent years, with 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%–2.20%, which has become a significant threat to public health.

[0003] Currently, lowering uric acid levels (including inhibiting uric acid production and promoting uric acid excretion) remains the core strategy for treating hyperuricemia. Xanthine oxidase (XOD) is a key enzyme in purine metabolism, and abnormal activity of XOD can lead to increased uric acid production. The uric acid transporter 1 (URAT1), as the main channel for uric acid reabsorption, is an important target for uricosuric drugs. The inhibition rates of XOD and URAT1 can be used to directly lower uric acid levels.

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

[0005] Therefore, there is an urgent need to develop a composition that combines highly efficient XOD and URAT1 dual inhibitory activity, has a simple composition, and high bioavailability to lower uric acid. Summary of the Invention

[0006] To address the aforementioned technical problems, this application provides a composition for lowering uric acid that can enhance the dual inhibitory activity of XOD and URAT1, has a simplified composition, and high bioavailability.

[0007] In a first aspect, this application provides a composition for lowering uric acid, the composition comprising the following raw materials in parts by weight: 10-30 parts of Phyllanthus emblica, 15-20 parts of Gardenia jasminoides, 15-25 parts of Astragalus membranaceus, and 35-50 parts of Chicory.

[0008] By adopting the above-mentioned technical solution, this application has a simple composition and achieves regulation of the "generation-excretion" dual pathway of uric acid metabolism, with high bioavailability of active ingredients. Specifically, Phyllanthus emblica has the effects of lowering blood uric acid levels, inhibiting gouty inflammation, and providing analgesia; Gardenia jasminoides can inhibit XOD activity and regulate the expression of uric acid transporters; Astragalus membranaceus can alleviate acute hyperuricemia by reducing uric acid production and providing renal protection, and promote uric acid excretion by regulating uric acid transporters; Chicory can reduce uric acid by lowering uric acid metabolic enzymes and key reaction enzymes, such as reducing the activity of xanthine oxidase, and can also achieve uric acid metabolism balance by inhibiting reabsorption and regulating the intestinal environment through renal and intestinal excretion. The synergistic combination of these four drugs significantly enhances the dual inhibitory activity against XOD and URAT1, thereby lowering uric acid levels.

[0009] Optionally, the composition comprises the following raw materials in parts by weight: 10 parts Phyllanthus emblica, 20 parts Gardenia jasminoides, 20 parts Chicoryrax praecox, and 50 parts Astragalus membranaceus.

[0010] Through the above technical solution, this application uses a specific ratio of Phyllanthus emblica, Gardenia jasminoides, Astragalus membranaceus, and Chicory, which can improve the dual inhibitory activity of XOD and URAT1, as well as the activity of total saponins, crude polysaccharides, total sugars, total flavonoids, and other active ingredients, as well as antioxidant and antiglycemic activities.

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

[0012] Optionally, the raw materials of the composition may 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, this 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 radicals, thereby inhibiting the production of uric acid caused by oxidative stress. Sea cucumber peptide can bind to the molybdenum pterin cofactor of 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 enhance the dual inhibitory activity of XOD and URAT1, the activity of total saponins, crude polysaccharides, total sugars, total flavonoids, and other active ingredients, as well as antioxidant and antiglycemic activities.

[0014] Optionally, the composition is obtained by fermentation of the raw material with Lactobacillus plantarum.

[0015] By adopting the above technical solution, 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.

[0016] Secondly, the present invention provides a method for preparing a composition for lowering uric acid, comprising the following steps:

[0017] S1. Pretreatment: Weigh the raw materials according to the proportion, crush them, and obtain Chinese medicine powder;

[0018] S2. Fermentation: Mix the Chinese medicine powder with sterile water evenly to obtain a mixture, then add Lactobacillus plantarum, and ferment at 35-40℃ for 12-72 hours for the first time, then cool down to 26-28℃ to continue fermentation.

[0019] S3. Purification: filtration, sterilization, evaporation and concentration.

[0020] Optionally, in S1, the pulverization mesh size is 60 to 80 mesh.

[0021] 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 mixture.

[0022] Optionally, in step S2, at least one of tuna peptide powder and sea cucumber peptide is added to the mixture, and the ratio of the initial fermentation time to the continued fermentation time is (0.5-2):1, with fermentation carried out in a shaker.

[0023] Optionally, in step S3, the sterilization temperature is 105–120°C, the evaporation temperature is 50–60°C, and the mixture is concentrated to 20–30% of its volume.

[0024] When the raw materials of the composition further include tuna peptide powder and / or sea cucumber peptide, the method for preparing the uric acid-lowering composition includes the following steps:

[0025] 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 mixed powder.

[0026] S2. Fermentation: Mix the powder mixture with sterile water to obtain a mixture, then add Lactobacillus plantarum. Ferment at 35-40℃ for 12-72 hours, then cool down to 26-28℃ to continue fermentation.

[0027] S3. Purification: filtration, sterilization, evaporation and concentration.

[0028] Optionally, in step S2, the weight ratio of sterile water to the mixed powder is (9-11):1.

[0029] By adopting the above technical solution, this application uses a two-stage temperature fermentation method, which can avoid the degradation of heat-sensitive components, improve the bioavailability of active ingredients, and thus enhance the dual inhibitory activity of XOD and URAT1, antioxidant activity, and anti-glycemic activity.

[0030] In summary, this application includes at least one of the following beneficial technical effects:

[0031] 1. This application has a simple composition, achieves regulation of the dual pathway of uric acid metabolism "generation-excretion", and has high bioavailability of active ingredients;

[0032] 1. This application uses a specific ratio of Phyllanthus emblica, Gardenia jasminoides, Astragalus membranaceus, and Chicory, which can improve the dual inhibitory activity of XOD and URAT1, as well as the activity of total saponins, crude polysaccharides, total sugars, total flavonoids, and other active ingredients, as well as antioxidant and antiglycemic activities.

[0033] 3. This application uses inoculation with Lactobacillus plantarum, which can improve the bioavailability of active ingredients such as total saponins, crude polysaccharides, total sugars, and total flavonoids;

[0034] 4. This application employs a two-stage temperature fermentation process, which avoids the degradation of heat-sensitive components, improves the bioavailability of active ingredients, and thereby enhances the dual inhibitory activity of XOD and URAT1, antioxidant activity, and antiglycemic activity. Attached Figure Description

[0035] Figure 1 This is a schematic diagram illustrating the effect of the composition of this application on URAT1 expression. A: URAT1 and β-actin protein bands; B: Relative expression level of URAT1. 1# represents the in vitro URAT1 expression in Comparative Example 1, and 2# represents the in vitro URAT1 expression in Example 1. ###p<0.001 vs Control, ***p<0.001 vs Model. Detailed Implementation

[0036] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0037] This application designs a composition for lowering uric acid, the composition comprising the following raw materials in parts by weight: 10-30 parts of Phyllanthus emblica, 15-20 parts of Gardenia jasminoides, 15-25 parts of Astragalus membranaceus, and 35-50 parts of Chicory.

[0038] The uric acid-lowering composition of this application is prepared by the following method, including the following steps:

[0039] S1. Pretreatment: Weigh the Chinese herbal raw materials according to the ratio, and pulverize them to 60-80 mesh to obtain Chinese herbal powder;

[0040] S2. Fermentation: Mix the Chinese herbal medicine powder with sterile water evenly. The weight ratio of sterile water to Chinese herbal medicine powder is (9-11):1 to obtain a mixture. Then add 3%-5% of the total volume of the mixture of Lactobacillus plantarum culture medium and place it in a shaker. Ferment for the first time at 35-40℃ for 12-72 hours, and then cool down to 26-28℃ to continue fermentation for 24-60 hours.

[0041] S3. Purification: Filtration, sterilization at 105-120℃, evaporation and concentration at 50-60℃, concentration to 20-30% of the volume of the mixed liquid.

[0042] All raw materials used in the embodiments of this application are commercially available, wherein:

[0043] Phyllanthus emblica, Anhui Mudan Tianxia Technology Co., Ltd.;

[0044] Gardenia, Anhui Mudan Tianxia Technology Co., Ltd.;

[0045] Astragalus, Anhui Mudan Tianxia Technology Co., Ltd.;

[0046] Chicory, Anhui Mudan Tianxia Technology Co., Ltd.;

[0047] Lactobacillus plantarum, Hebei Yiran Biotechnology Co., Ltd.;

[0048] Tuna peptide powder, Yantai Jia Hui Marine Biotechnology Co., Ltd.;

[0049] Sea cucumber peptides, Yantai Jia Hui Marine Biotechnology Co., Ltd.

[0050] Testing items and testing methods

[0051] 1. Determination of in vitro XOD inhibitory activity and half-maximal inhibitory concentration (IC50):

[0052] Prepare 0.5 U / mL xanthine oxidase solution and 2.0 μmol / L xanthine solution using 0.2 mol / L phosphate buffer. Add 50 μL of xanthine oxidase solution to 200 μL of the test solution and mix. Incubate at 25°C for 15 min, then add 400 μL of xanthine solution and continue incubation for 20 min. Finally, add 50 μL of 1 mol / L hydrochloric acid solution to terminate the reaction. Measure the absorbance (A3) at 290 nm and calculate the XOD activity inhibition rate. Also, measure the absorbance (A0) without the test solution and xanthine oxidase solution, the absorbance (A1) without the test solution, and the absorbance (A2) without the xanthine oxidase solution. XOD activity inhibition rate = [(A1-A0)-(A3-A2)] / (A1-A0)*100%. The test solution was diluted with 55% ethanol at 2, 4, 8, and 16 times, and the absorbance was measured according to the above method. A dose-response curve was plotted to obtain the IC50 value.

[0053] 2. In vitro URAT1 inhibitory activity assay:

[0054] Human renal tubular epithelial cells (HK-2) were divided into groups of 7.2 × 10⁻⁶. 5 Cells were seeded per well in 6cm dishes and administered drug treatment 24 hours later. 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. Proteins were extracted from each treatment group after 48 hours of culture. β-actin was used as an internal control, and the effect of each group on URAT1 expression was detected by Western blotting. The experiment was repeated three times. URAT1 activity inhibition rate = (URAT1 expression level in the model group - URAT1 expression level in the experimental group) / URAT1 expression level in the model group * 100%.

[0055] 3. Determination of active ingredients (total saponins, crude polysaccharides, total sugars, total flavonoids)

[0056] 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 a standard, and a standard curve was plotted. The total saponin content of the test solution was determined at 543 nm.

[0057] 3.2 Determination of crude polysaccharide content: The crude polysaccharide content of the test solution was determined in accordance with the entry-exit inspection and quarantine industry standard SN / T 4260—2015 "Determination of crude polysaccharides in exported plant-derived foods by phenol-sulfuric acid method".

[0058] 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 plotted. The total sugar content of the test solution was measured at 490 nm.

[0059] 3.4 Determination of Total Flavonoid Content: The total flavonoid content of the test solution was determined using the aluminum nitrate-sodium hydroxide colorimetric method. Rutin was used as a standard, and a standard curve was plotted. The total flavonoid content of the test solution was measured at 510 nm.

[0060] 4. Determination of antioxidant activity

[0061] 4.1 Hydroxyl radical scavenging rate: Mix 50 μL of the test solution, 50 μL of 2 mmol / L ferrous sulfate solution, and 50 μL of 6 mmol / L hydrogen peroxide solution thoroughly. Let stand for 10 min, add 50 μL of salicylic acid solution, heat at 37℃ for 15 min, and then measure the absorbance A1 at 510 nm. The absorbance without the test solution is taken as A0, and the absorbance without hydrogen peroxide is taken as A2. The experiment is repeated 3 times. Hydroxyl radical scavenging rate = [(A1-A2) / A0]*100%;

[0062] 4.2 DPPH Scavenging Rate: Mix 50 μL of the test solution and 150 μL of DPPH solution thoroughly, heat at 37℃ for 30 min, and measure the absorbance (A1) at 517 nm. The absorbance measured with an equal volume of 70% ethanol instead of the test solution is designated as A2, and the absorbance measured without adding the test solution is designated as A0. The experiment was repeated 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 ABTS solution thoroughly, react at room temperature for 30 min, and measure the absorbance (A1) at 735 nm. The absorbance measured with an equal volume of 70% ethanol instead of the test solution is designated as A2, and the absorbance measured without adding the test solution is designated as A0. The experiment was repeated three times. ABTS scavenging rate = [1 - (A1 - A2) / A0] * 100%.

[0063] 5. Antiglycemic activity assay

[0064] The inhibition rate of the test solution on α-glucosidase activity was determined using the Lineweaver-Burk double reciprocal method. Specific Implementation

[0066] Preparation Example 1

[0067] Preparation of Lactobacillus plantarum culture medium:

[0068] Live Lactobacillus plantarum was cultured anaerobically at 37°C for 48 hours; serially diluted and plated to obtain single colonies; single colonies were picked and cultured for 12 hours, and then inoculated into MRS liquid medium at a 2% inoculum and cultured at 37°C for 24 hours.

[0069] Example 1

[0070] Weigh out 1g of Phyllanthus emblica, 2g of Gardenia jasminoides, 2g of Astragalus membranaceus, and 5g of Chicory, and pulverize them to 60 mesh to obtain a traditional Chinese medicine powder. Mix the powder with sterile water at a weight ratio of 10:1 to obtain a mixture. Then, add 3% of the total volume of the mixture to the Lactobacillus plantarum culture obtained in Preparation Example 1. Place the mixture in a shaker and ferment at 37°C for 12 hours. Then, cool the mixture to 28°C and continue fermenting for 60 hours. Filter the mixture, sterilize it at 120°C, and evaporate and concentrate it at 60°C to 20% of the volume of the mixture to obtain a fermentation broth that lowers uric acid.

[0071] Example 2

[0072] Weigh out 2g of Phyllanthus emblica, 1.5g of Gardenia jasminoides, 2.5g of Astragalus membranaceus, and 4g of Chicory, and pulverize them to 80 mesh to obtain a traditional Chinese medicine powder. Mix the powder with sterile water at a weight ratio of 9:1 to obtain a mixture. Then, add 5% of the total volume of the mixture to the Lactobacillus plantarum culture obtained in Preparation Example 1. Place the mixture in a shaker and ferment at 40°C for 12 hours. Then, cool the mixture to 27°C and continue fermenting for 60 hours. Filter the mixture, sterilize it at 110°C, and evaporate and concentrate it at 55°C to 25% of the volume of the mixture to obtain a fermentation broth that lowers uric acid.

[0073] Example 3

[0074] Weigh out 3g of Phyllanthus emblica, 2g of Gardenia jasminoides, 1.5g of Astragalus membranaceus, and 3.5g of Chicory, and pulverize them to 80 mesh to obtain a traditional Chinese medicine powder. Mix the powder with sterile water at a weight ratio of 11:1 to obtain a mixture. Then, add 5% of the total volume of the mixture to the Lactobacillus plantarum culture obtained in Preparation Example 1. Place the mixture in a shaker and ferment at 35°C for 12 hours. Then, cool the mixture to 26°C and continue fermenting for 60 hours. Filter the mixture, sterilize it at 105°C, and evaporate and concentrate it at 50°C to 30% of the volume of the mixture to obtain a fermentation broth that lowers uric acid.

[0075] The fermentation broths of the uric acid-lowering compositions obtained in Examples 1-3 were subjected to in vitro XOD inhibition activity, IC50 value, in vitro URAT1 inhibition activity, active ingredients (total saponins, crude polysaccharides, total sugars, and total flavonoids), antioxidant activity, and antiglycemic activity determination. The test results are shown in Table 1.

[0076] Table 1. Determination of the uric acid reduction and bioactivity of the fermentation broths from Examples 1-3.

[0077]

[0078]

[0079] As shown in Examples 1-3 and Table 1, the uric acid-lowering composition obtained in this application was obtained by fermentation of Phyllanthus emblica, Gardenia jasminoides, Astragalus membranaceus, and Chicory in a specific ratio. The pH value decreased after fermentation, indicating successful fermentation. This application uses a specific ratio of Phyllanthus emblica, Gardenia jasminoides, Astragalus membranaceus, and Chicoryry, which can improve the dual inhibitory activity of XOD and URAT1, while also increasing the levels of active ingredients such as total saponins, crude polysaccharides, total sugars, and total flavonoids, as well as antioxidant and antiglycemic activities.

[0080] Comparative Example 1

[0081] The difference between this comparative example and Example 1 is that in this comparative example, the Chinese herbal raw materials (1g of Phyllanthus emblica, 2g of Gardenia jasminoides, 2g of Astragalus membranaceus, and 5g of Chicory) from Example 1 were added to sterile water at a material-to-liquid ratio of 1:10, heated in a water bath, boiled and extracted for 1.5 hours, filtered and the filtrate was collected; the above steps were repeated 3 times, the filtrates were combined, evaporated and concentrated to obtain an extract equal in volume to the fermentation broth of the composition for lowering uric acid in Example 1, while in this comparative example, an equal volume of the fermentation broth of the composition was replaced with the extract.

[0082] Comparative Example 2

[0083] The difference between this comparative example and Example 1 is that this comparative example was obtained by fermentation at 28°C for 72 hours.

[0084] Comparative Example 3

[0085] The difference between this comparative example and Example 1 is that the mass of Phyllanthus emblica is replaced with Gardenia jasminoides in this comparative example.

[0086] Comparative Example 4

[0087] The difference between this comparative example and Example 1 is that chicory is not added to this comparative example.

[0088] Comparative Example 5

[0089] The difference between this comparative example and Example 1 is that the gardenia and other fruits are replaced with amla in this comparative example.

[0090] Comparative Example 6

[0091] The difference between this comparative example and Example 1 is that Astragalus membranaceus is not added to this comparative example.

[0092] The fermentation broths of the uric acid-lowering compositions obtained in Example 1 and Comparative Examples 1-5 were subjected to in vitro XOD inhibition activity, IC50 value, in vitro URAT1 inhibition activity, active ingredients (total saponins, crude polysaccharides, total sugars, and total flavonoids), antioxidant activity, and antiglycemic activity determination. The test results are shown in Table 2.

[0093] Table 2. Determination of uric acid level reduction and bioactivity of fermentation broths from Examples 1 and Comparative Examples 1-5.

[0094]

[0095]

[0096] As shown in Example 1, Comparative Example 1, and Table 2, compared to the extract, the IC50 value of the composition decreased, while the XOD activity inhibition rate, URAT1 activity inhibition rate, content of active ingredients (total saponins, crude polysaccharides, total sugars, and total flavonoids), antioxidant activity, and antiglycemic activity increased. This is mainly because fermentation of *Lactobacillus plantarum* can produce organic acids, leading to a decrease in acidity. Furthermore, by decomposing plant cell walls, it releases active ingredients such as saponins, polysaccharides, and flavonoids, thereby improving bioavailability and thus enhancing the dual inhibitory activity of XOD and URAT1, antioxidant activity, and antiglycemic activity. This application utilizes fermentation treatment of the traditional Chinese medicine composition, which can significantly improve the utilization rate of bioactive ingredients, thereby enhancing the dual inhibitory activity of XOD and URAT1, antioxidant activity, and antiglycemic activity.

[0097] As shown in Example 1, Comparative Example 2, and Table 2, during continuous fermentation at 28°C, the cell growth is slow and the metabolic activity is insufficient, resulting in a reduction in the dissolution of primary metabolites (such as crude polysaccharides and total sugars) and a decrease in the enzymatic conversion efficiency of secondary metabolites (such as saponins and flavonoids). This application employs a two-stage temperature fermentation. The high-temperature stage at 37°C promotes the rapid proliferation of Lactobacillus plantarum, accelerates cell wall decomposition, and releases primary metabolites such as polysaccharides and saponins. The low-temperature stage at 28°C avoids the degradation of heat-sensitive components (flavonoids, saponins, etc.). Maintaining the two-stage temperature fermentation can balance metabolic efficiency and component stability, thereby significantly enhancing the dual inhibition effect of XOD and URAT1, and improving antioxidant and antiglycemic activities.

[0098] As shown in Example 1, Comparative Example 3, and Table 2, replacing Phyllanthus emblica with Gardenia jasminoides, although Gardenia jasminoides itself has an inhibitory effect on XOD, disrupts the original formula's dual-target synergistic mechanism of "production-excretion" (Phyllanthus emblica is responsible for excretion regulation, while Gardenia jasminoides mainly targets production inhibition), resulting in a significant decrease in the URAT1 inhibition rate. Furthermore, the absence of Phyllanthus emblica directly reduces the amount of active ingredients such as saponins and flavonoids, weakening antioxidant and antiglycemic activities, and slightly decreasing the content of crude polysaccharides and total sugars.

[0099] As can be seen from Example 1, Comparative Example 4 and Table 2, without the addition of chicory, the fermentation broth of the resulting 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 antiglycemic activities.

[0100] As shown in Example 1, Comparative Example 5, and Table 2, after replacing Gardenia with Phyllanthus emblica, the fermentation broth of the composition lost its direct inhibitory effect on XOD (Gardenia as the core target), resulting in a significant decrease in the XOD activity inhibition rate. Although Phyllanthus emblica can partially compensate for the URAT1 inhibitory effect through excretion, the overall synergistic mechanism is disrupted, and the URAT1 inhibition rate is still slightly reduced. In addition, the total flavonoids and antioxidant activity are significantly reduced due to the absence of Gardenia.

[0101] As can be seen from Example 1, Comparative Example 6 and Table 2, without the addition of Astragalus membranaceus, the XOD activity inhibition rate decreased slightly, but the URAT1 activity inhibition rate decreased significantly, and the total saponin content was also significantly reduced, and the antioxidant and anti-glycemic activities were weakened.

[0102] As can be seen from Example 1, Comparative Examples 3-6 and Table 2, this application uses Phyllanthus emblica, Gardenia jasminoides, Astragalus membranaceus and Chicory in synergistic combination to achieve dual high-efficiency inhibition of XOD and URAT1, while taking into account highly active ingredients (total saponins, polysaccharides, flavonoids) and comprehensive bioactivity (antioxidant, antiglycemic).

[0103] Example 4

[0104] Weigh out 1g of Phyllanthus emblica, 2g of Gardenia jasminoides, 2g of Astragalus membranaceus, and 5g of Chicory, and pulverize them to 60 mesh to obtain a traditional Chinese medicine powder. Add 0.4g of tuna peptide powder to obtain a mixed powder. Mix the mixed powder with sterile water at a weight ratio of 10:1 to obtain a mixed solution. Then add 3% of the total volume of the mixed solution of *Lactobacillus plantarum* culture obtained in Preparation Example 1. Place the mixture in a shaker and ferment at 37°C for 12 hours. Then cool the temperature to 28°C and continue fermenting for 60 hours. Filter the mixture, sterilize it at 120°C, and evaporate and concentrate it at 60°C to 20-30% of the volume of the mixed solution to obtain a fermentation broth that lowers uric acid.

[0105] Example 5

[0106] The difference between this embodiment and embodiment 4 is that in this embodiment, the tuna peptide powder is replaced with sea cucumber peptide.

[0107] Example 6

[0108] The difference between this embodiment and embodiment 4 is that in this embodiment, 0.4g of tuna peptide powder is replaced with 0.2g of tuna peptide powder and 0.2g of sea cucumber peptide.

[0109] The fermentation broths of the uric acid-lowering compositions obtained in Examples 1 and 4-6 were subjected to in vitro XOD inhibition activity, IC50 value, in vitro URAT1 inhibition activity, active ingredients (total saponins, crude polysaccharides, total sugars, and total flavonoids), antioxidant activity, and antiglycemic activity determination. The test results are shown in Table 3.

[0110] Table 3. Determination of the uric acid reduction and bioactivity of the fermentation broth compositions from Examples 1 and 4-6.

[0111]

[0112]

[0113] As shown in Examples 1, 4-6 and Table 3, adding tuna peptide powder alone can significantly improve the URAT1 inhibition rate and DPPH scavenging rate, adding sea cucumber peptide alone can significantly improve the XOD activity inhibition rate and IC50 value, and the combined use of tuna peptide powder and sea cucumber peptide can improve the dual inhibitory activity of XOD and URAT1, the activity of total saponins, crude polysaccharides, total sugars, total flavonoids and other active ingredients, as well as antioxidant activity and anti-glycemic activity.

[0114] Example 7

[0115] The difference between this embodiment and embodiment 6 is that this embodiment uses fermentation at 37°C for 24 hours, followed by cooling to 28°C and continuing fermentation for another 48 hours.

[0116] Example 8

[0117] The difference between this embodiment and embodiment 6 is that this embodiment uses fermentation at 37°C for 48 hours, followed by cooling to 28°C and continuing fermentation for another 24 hours.

[0118] Example 9

[0119] The difference between this embodiment and embodiment 6 is that this embodiment is obtained by fermentation at 37°C for 72 hours.

[0120] The fermentation broths of the uric acid-lowering compositions obtained in Examples 6 and 7-9 were subjected to in vitro XOD inhibition activity, IC50 value, in vitro URAT1 inhibition activity, active ingredients (total saponins, crude polysaccharides, total sugars, and total flavonoids), antioxidant activity, and antiglycemic activity determination. The test results are shown in Table 4.

[0121] Table 4. Determination of uric acid reduction and bioactivity of the fermentation broth compositions from Examples 6 and 7-9.

[0122]

[0123]

[0124] As shown in Examples 5, 7-9, and Table 4, this application employs a two-stage temperature fermentation process. The first 24 hours of high-temperature fermentation (37°C) promotes rapid microbial growth and the accumulation of primary metabolites (such as polysaccharides and saponins). The subsequent 48 hours of low-temperature fermentation (28°C) slows down the microbial growth rate, prolongs the synthesis cycle of secondary metabolites (such as saponins and flavonoids), and prevents the degradation of peptides and flavonoids. Excessive high-temperature fermentation (37°C) can lead to metabolic imbalance, degradation or consumption of active ingredients, decomposition of crude polysaccharides into monosaccharides, and a decline in other indicators. This application uses specific temperatures and fermentation times, which can significantly increase the levels of active ingredients such as total saponins, crude polysaccharides, total sugars, and total flavonoids without reducing the inhibitory activity of XOD and URAT1.

[0125] The embodiments described in this specific implementation are preferred embodiments of this application and are not intended to limit the scope of protection of this application. Therefore, any simple modifications, equivalent substitutions, and improvements made based on the technical solutions of this application should be covered within the scope of protection of this application.

Claims

1. A composition for reducing uric acid, characterized by comprising, The composition is made of the following raw materials by weight: Emblica 10-30 parts, Gardenia 15-20 parts, Astragalus 15-25 parts, and Chichorium 35-50 parts; the composition is obtained by fermenting the raw materials by Lactobacillus plantarum.

2. The composition for reducing uric acid according to claim 1, wherein The composition is made of the following raw materials by weight: Emblica 10 parts, Gardenia 20 parts, Astragalus 20 parts, and Chichorium 50 parts.

3. The composition for reducing uric acid according to claim 1, wherein The raw materials of the composition can further add 2 parts by weight of tuna peptide powder and / or 2 parts by weight of sea cucumber peptide.

4. A preparation method of the uric acid-lowering composition of any one of claims 1-3, comprising the following steps: S1, pretreatment: weighing the raw materials according to the ratio, crushing to obtain raw material powder; S2, fermentation: mixing the raw material powder with sterile water uniformly, the weight ratio of sterile water to raw material powder is (9-11):1, to obtain a mixed solution; then adding Lactobacillus plantarum culture solution, the volume of Lactobacillus plantarum culture solution is 3%-5% of the total volume of the mixed solution, after the first fermentation at 35-40℃ for 12-72h, the temperature is lowered to 26-28℃ for continuous fermentation; S3, purification: filtering, evaporating and concentrating, sterilizing.

5. The method of claim 4, wherein the composition is prepared by mixing the uric acid-lowering agent and the pharmaceutically acceptable carrier. In the step S1, the crushing mesh number is 60-80.

6. The method of claim 4, wherein the composition is prepared by mixing the xanthine oxidase inhibitor and the uricosuric agent. In the step S2, the ratio of the first fermentation time to the continuous fermentation time is (0.5-2):1, and the fermentation is carried out in a shaking bed.

7. The method of claim 4, wherein the composition is prepared by mixing the xanthine oxidase inhibitor and the uricosuric agent. In the step S3, the sterilization temperature is 105-120℃, and the evaporation temperature is 50-60℃.

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