Composition with effects of reducing uric acid and protecting kidney and application thereof

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a composition with uric acid reducing and kidney protecting effects and application thereof. The preparation method comprises the following steps: firstly, carrying out alcohol extraction, column chromatography purification and concentration to obtain pueraria polyphenol from pueraria; carrying out deproteinization and purification through an ultrasonic-assisted water extraction method, and extracting polygonatum polysaccharide from polygonatum cyrtonema; the radix puerariae polyphenol and the rhizoma polygonati polysaccharide are mixed according to the mass ratio of 2: (2-10), the composition with the uric acid lowering and kidney protecting effects is obtained, through the synergistic effect of the rhizoma polygonati polysaccharide and the radix puerariae polyphenol in the composition, the uric acid lowering effect of the radix puerariae polyphenol is improved, meanwhile, inflammatory response can be inhibited, kidney pathological injuries are relieved, and therefore the kidney is protected; the composition has potential application value in preparation of uric acid reducing drugs, functional food and the like.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical technology, and particularly relates to a composition with the effects of reducing uric acid and protecting the kidneys and its application. Background Art

[0002] Hyperuricemia (HUA) is a metabolic disease caused by purine metabolism disorders or / and reduced uric acid excretion, characterized by excessive uric acid levels in the blood. Long-term hyperuricemia can cause urate crystals to deposit in joint cavities and soft tissues, leading to gout; HUA can also directly cause a series of metabolic syndromes such as cardiovascular diseases, diabetes, and chronic kidney diseases, seriously threatening human health. Most mammals contain uricase, which can oxidize and degrade uric acid into a soluble compound, allantoin. However, during human evolution, the gene encoding uricase mutated, resulting in the absence of uricase; uric acid maintains a balance between production in the liver and excretion through the kidneys and intestines to ensure the homeostasis of uric acid levels in the human body. When the balance between uric acid production and excretion is disrupted, the body's uric acid levels will increase, leading to hyperuricemia.

[0003] In the clinical treatment field of hyperuricemia, current treatment methods are mainly dominated by Western medicines. However, while these Western medicines play a role in reducing uric acid, they are often accompanied by significant side effects that cannot be ignored. After many patients take Western medicines for a long time, it is relatively common for liver and kidney functions to be damaged, which not only poses a new threat to the health of patients but also limits the application effect of Western medicines in long-term treatment. In addition, although existing traditional Chinese medicine compositions provide treatment options to a certain extent, due to the complex components of traditional Chinese medicine, if the compatibility is not accurately controlled during use, it may also cause significant side effects due to the inherent toxicity of certain components or drug interactions, and even interfere with normal physiological functions, causing potential damage to important organs such as the liver and kidneys. Therefore, developing a highly efficient, safe, and long-term consumable product for reducing uric acid is of great significance for the research and development of uric acid-lowering drugs. Summary of the Invention

[0004] To solve the above problems, the present invention provides a composition with the effects of reducing uric acid and protecting the kidneys and its application. The composition is obtained by mixing puerarin polyphenol and polygonatum polysaccharide in a mass ratio of 2:2 to 10, and has the characteristics of natural safety, low toxicity, and low irritation. Through animal experiments, it is found that polygonatum polysaccharide and puerarin polyphenol in the composition not only improve the uric acid-lowering efficacy of puerarin polyphenol through a synergistic effect, but also can inhibit inflammatory reactions and reduce kidney pathological damage, thereby protecting kidney function.

[0005] To achieve the above object, the specific technical solution of the present invention is as follows: The first aspect of the present invention provides a composition with the effects of reducing uric acid and protecting the kidneys, which is characterized in that the composition is prepared by mixing puerarin polyphenols and polygonatum polysaccharides in a mass ratio of 2:2 to 10.

[0006] Further, the puerarin polyphenols are obtained by extracting crude polyphenols from pueraria through alcohol extraction, purifying by column chromatography, and concentrating.

[0007] Further, the polygonatum polysaccharides are obtained by extracting crude polysaccharides from polygonatum cyrtonema through ultrasonic-assisted water extraction and deproteinizing and purifying.

[0008] The second aspect of the present invention provides an application of the above-mentioned composition in the preparation of a drug for treating hyperuricemia, and the drug for treating hyperuricemia is composed of the composition and excipients.

[0009] Further, the dosage form of the drug for treating hyperuricemia is dispersible tablets, soft capsule agents, granule agents, powder agents, dripping pills, gel agents or oral liquid preparations.

[0010] Further, the drug for treating hyperuricemia is a drug for reducing uric acid content, reducing urea content or alleviating renal pathological damage.

[0011] The third aspect of the present invention provides an application of the above composition in the preparation of a functional food, and the functional food is a jelly-type gel or a gummy gel.

[0012] Further, the gummy gel is composed of, by mass: 0.7 parts to 1.5 parts of puerarin polyphenols, 1.0 parts to 3.5 parts of polygonatum polysaccharides, 3.75 parts of gelatin, 1.25 parts of carrageenan, 1.0 part of cyclodextrin, 1 part of sodium carboxymethylcellulose, 2 parts to 4 parts of konjac gum, and 0.1 part of KCl.

[0013] Further, the jelly gel is composed of, by mass: 0.7 parts to 1.5 parts of puerarin polyphenols, 1.0 parts to 3.5 parts of polygonatum polysaccharides, 3.75 parts of gelatin, 1.25 parts of carrageenan, 1.0 part of cyclodextrin, 1 part of sodium carboxymethylcellulose, 0.5 parts to 2 parts of pectin, 0.1 part of KCl, and 0.1 part of CaCl2.

[0014] The formula of the jelly-type gel or gummy gel contains no sucrose and no glucose; cyclodextrin is used to encapsulate puerarin polyphenols, which increases the water solubility of puerarin polyphenols and also eliminates the bitter taste of puerarin polyphenols; adding sodium carboxymethylcellulose increases the viscosity of the gel and plays a flocculation role.

[0015] Compared with the prior art, the beneficial effects of the present invention are as follows: The present invention obtained pueraria polyphenols from kudzu root through alcohol extraction, column chromatography purification and concentration; extracted polygonatum polysaccharides from polygonatum cyrtonema hua through ultrasonic-assisted water extraction and protein removal purification; then mixed pueraria polyphenols and polygonatum polysaccharides in a mass ratio of 2:2 to 10 to obtain a composition with the effects of reducing uric acid and protecting the kidneys. This composition has the effects of reducing uric acid and protecting the kidneys, and has potential application value in the preparation of drugs for reducing uric acid, functional foods, etc.

[0016] (1) Synergistic effect and improved bioavailability: The polygonatum polysaccharides and pueraria polyphenols in the composition provided by the present invention have a significant synergistic effect in reducing uric acid and protecting renal function. Pueraria polyphenols have poor solubility, while polygonatum polysaccharides are a good drug-loading sustained-release agent, which can make pueraria polyphenols have better hydrophilicity and sustained-release effects, enhancing the bioavailability of pueraria polyphenols and achieving good experimental results at a lower dose. At the same time, polygonatum polysaccharides also exert their own biological functions, combining drug and adjuvant. It simplifies the ingredients and improves the therapeutic effect, ensuring significant uric acid reduction and kidney protection effects at a lower dose.

[0017] (2) High-efficiency effect of reducing uric acid and protecting the kidneys: The HE pathological staining results of the kidneys of mice in the present invention and the levels of serum inflammatory factors show that the renal injury of mice in the hyperuricemia group is obvious. Although allopurinol has a good effect of reducing uric acid, it does not improve the renal injury of mice, and the renal inflammatory factors increase significantly, indicating that the toxic and side effects of western medicine cannot be ignored. However, after 14 days of administration of the PPE composition, the serum uric acid level of hyperuricemic mice decreased significantly and could be reduced to the normal level; the contents of creatinine and urea in the serum were not significantly different from those in the normal group; the levels of TNF-α and IL-1β in the kidney tissue were significantly decreased compared with the hyperuricemia model group, and the renal urate transporter 1 (URAT1) was also down-regulated to the normal level, indicating that the interaction between polygonatum polysaccharides and pueraria polyphenols in the composition of the present invention not only improves the uric acid-lowering efficacy of pueraria polyphenols, but also inhibits the inflammatory response and reduces the renal pathological injury, thereby protecting renal function.

[0018] (3) Low toxic and side effects and natural safety: Compared with traditional western medicine, this composition has low toxic and side effects and natural safety, avoiding further damage to the kidneys. The polygonatum polysaccharides and pueraria polyphenols in the PPE composition are both derived from natural medicinal materials that are both medicine and food, with natural safety, low toxicity and low irritation, and are suitable for long-term use by patients with hyperuricemia, providing a good basis for the development of new health foods and natural drugs.

[0019] (4) Great application potential: Animal experiments in the present invention show that the composition provided by the present invention has a high-efficiency effect of reducing uric acid and protecting the kidneys and can be used to prepare drugs for reducing uric acid; based on this composition, the present invention developed functional food gel gummies and gel jellies, providing an experimental basis for the development of characteristic health products with potential new functions of reducing uric acid.

[0020] In summary, by reasonably compounding polygonatum polysaccharide and pueraria polyphenol, the present invention not only improves the effect of reducing uric acid, but also significantly improves the kidney protection function, has good safety and bioavailability, provides an innovative solution for the development of therapeutic drugs and health foods for hyperuricemia, and has important clinical and market application values. Brief Description of the Drawings

[0021] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for the description of the embodiments or the prior art. Obviously, the following drawings are only some embodiments of the present invention. For those of ordinary skill in the art, other drawings can be obtained based on these drawings without creative efforts.

[0022] Figure 1 Effect of PPE on blood uric acid, creatinine, and urea contents in hyperuricemic mice; the detection indexes corresponding to Figures A, B, and C are uric acid, creatinine, and urea contents in sequence; compared with the normal group # is p <0.05, ## is p <0.01; compared with the model group, * is p <0.05, ** is p <0.01.

[0023] Figure 2 Effect of PPE on TNF-α, IL-1β, and IL-6 in the kidneys of hyperuricemic mice; the detection indexes corresponding to Figures A, B, and C are TNF-α, IL-1β, and IL-6 levels in sequence; compared with the normal group # is p <0.05, ## is p <0.01; compared with the model group, * is p <0.05, ** is p <0.01.

[0024] Figure 3 Microscopic pathological photos of renal tissues of mice in each group; Figures A, B, C, and D are microscopic pathological photos of renal tissues of mice in the normal group, hyperuricemia model group, positive control group, and PPE group in sequence; red arrow: glomerulus; blue arrow: renal capsule; green arrow: proximal tubule; black arrow: distal tubule.

[0025] Figure 4Effect of PPE on the expression level of URAT1 protein in the kidneys of hyperuricemia mice; Figure A is the WB image of the expression level of URAT1 protein. The upper row of bands is the URAT1 protein, and the lower row of bands is the internal reference protein. The samples of each row of bands from left to right are the corresponding protein contents of the normal group, hyperuricemia model group, positive control group, and PPE group; Figure B is the statistical chart of the expression level of URAT1 protein in Figure A; compared with the normal group ## is p <0.01; compared with the model group, * is p <0.05, ** is p <0.01.

[0026] Figure 5 is the texture curve diagram of gummy bears; Figure A is the texture curve diagram of RPPE gummy bears; Figure B is the texture curve diagram of PPE gummy bears.

[0027] Figure 6 is the texture curve diagram of jelly; Figure A is the texture curve diagram of RPPE jelly; Figure B is the texture curve diagram of PPE jelly. Specific embodiments

[0028] The specific embodiments of the present invention will be described in detail below, but it should be understood that the protection scope of the present invention is not limited by the specific embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts belong to the protection scope of the present invention. The experimental methods described in the embodiments of the present invention are all conventional methods unless otherwise specified. The materials, reagents, etc. used in the following embodiments can be obtained from commercial channels unless otherwise specified.

[0029] Polygonatum cyrtonema Hua in the present invention was purchased from the Polygonatum cyrtonema Hua planting base in Jinhua City, Zhejiang Province, and Pueraria lobata was purchased from Xi'an, Shaanxi; male Kunming mice were purchased from Tongxiang Branch of Zhejiang Vital River Laboratory Animal Technology Co., Ltd., certificate number: 20230525Abzz0600999711; allopurinol was purchased from Shanghai Macklin Biochemical Co., Ltd.; UA, Cr, and Urea kits were purchased from Shenzhen Mindray Bio-Medical Electronics Co., Ltd.; mouse tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β), and interleukin 6 (IL-6) enzyme-linked immunosorbent assay (ELISA) quantitative detection kits were purchased from Quanzhou Ruixin Biotechnology Co., Ltd., and HRP-labeled goat anti-rabbit Anti-URAT1 (primary antibody, Cat No: 14937-1-AP), Anti-ACTIN (primary antibody, Cat No.81115-1-RR), and secondary antibody (Cat No: SA00001-2) were all purchased from Wuhan Sanying Biotechnology Co., Ltd.

[0030] Hyperuricemia (HUA) is a metabolic disease caused by purine metabolism disorder or reduced uric acid excretion, characterized by excessive blood uric acid levels, which can trigger gout and various metabolic syndromes. Due to the evolutionary absence of uricase in humans, the imbalance between uric acid production and excretion easily leads to this disease. Currently, the clinical treatment for reducing uric acid mainly relies on Western medicine, but long-term use of Western medicine often damages the liver and kidneys of patients, restricting its application effect. Although existing traditional Chinese medicine compositions provide certain treatment options, their ingredients are complex. If the compatibility is not precisely controlled, it is easy to cause significant toxic and side effects due to the inherent toxins of the ingredients or drug interactions, threatening important organs such as the liver and kidneys. Therefore, developing a highly effective, safe, and long-term ingestible product for reducing uric acid is of great significance for the research and development of uric acid-lowering drugs.

[0031] The present invention provides a composition with the efficacy of reducing uric acid and protecting the kidneys and its application. Firstly, a puerarin polyphenol was obtained from Pueraria lobata through alcohol extraction, column chromatography purification, and concentration; a polygonatum polysaccharide was extracted from Polygonatum cyrtonema through ultrasonic-assisted water extraction and protein removal purification; then, the puerarin polyphenol and polygonatum polysaccharide were mixed at a mass ratio of 2:2 - 10 to obtain a composition with the efficacy of reducing uric acid and protecting the kidneys. Through animal experiments, it was found that the polygonatum polysaccharide and puerarin polyphenol in this composition not only improved the uric acid-lowering efficacy of puerarin polyphenol through synergistic effects, but also inhibited inflammatory reactions and reduced kidney pathological damage, thereby protecting kidney function. And based on this composition, the present invention developed a functional jelly candy and jelly with high moisture content, low sweetness, and low calories.

[0032] Example 1: Preparation of a composition with the efficacy of reducing uric acid and protecting the kidneys 1. Extraction and purification of polygonatum polysaccharide Extraction of polygonatum polysaccharide: Take 300 g of Polygonatum cyrtonema, cut it into slices, place it in a blender to break it up, add deionized water at a solid-to-liquid ratio of 1 g:10 mL, use an ultrasonic processor at 40 HZ, with a total ultrasonic time of 100 min. After ultrasonic treatment, filter by suction. Take the filter residue and add an equal amount of deionized water again, repeat the ultrasonic extraction 3 times, filter by suction. Combine the filtrates obtained from the three ultrasonic treatments, centrifuge at 4°C and 10000 rpm for 5 min, take the supernatant, concentrate it by reduced pressure rotary evaporation, add 4 times the volume of absolute ethanol, let it stand in a 4°C refrigerator overnight for precipitation, and centrifuge to obtain the precipitate, namely crude polygonatum polysaccharide (RPCP).

[0033] Purification of polygonatum polysaccharide: Dissolve the crude polygonatum polysaccharide in water, add neutral protease for hydrolysis, with the enzyme addition amount being 3.0% of the total volume of the polysaccharide solution, stir and mix evenly, and react in a 45°C constant temperature water bath for 1 h (stir at low speed every 10 min). Centrifuge at 10000 rpm for 5 min, take the supernatant, and perform membrane separation using an ultrafiltration membrane module with a cut-off relative molecular weight of 1 kDa. Freeze-dry the retentate to obtain pure white polygonatum polysaccharide (PCP).

[0034] The polysaccharide content was determined by the anthrone method. The results showed that the total content of crude polysaccharides of Polygonatum sibiricum obtained by ultrasonic-assisted water extraction was 89.5 g, accounting for 29.8% of the mass of Polygonatum sibiricum. After protein removal, 53.7 g of Polygonatum sibiricum polysaccharide was obtained.

[0035] 2. Extraction, concentration, and column chromatography purification of puerarin polyphenols Ultrasonic-assisted extraction of crude puerarin polyphenols: Add 250 g of pueraria powder to 1500 mL of 80% ethanol and ultrasonically crush (20 kHz, 100 min), then filter. Add 1500 mL of 80 v / v% ethanol to the filter residue and ultrasonically crush for another 100 min. Combine the two filtrates and concentrate under reduced pressure at 50 °C to obtain crude puerarin polyphenols (RPLP) for column chromatography.

[0036] Treatment of the resin: Macroporous resin D-101 was soaked in 95% ethanol for 24 h, washed with water until there was no alcohol smell, soaked in 5 v / v% HCl for 4 h, and washed with water until neutral; then soaked in 5 v / v% NaOH for 4 h and washed with water until neutral for standby.

[0037] Load the treated macroporous resin D-101 into the chromatography column and make the liquid level just cover the macroporous resin. Measure the mass of the remaining macroporous resin and the volume of the remaining water.

[0038] The polyphenol solid obtained by extraction and concentration was dissolved in water, stirred evenly, loaded onto the column at a flow rate of 1 mL / min, and the effluent was collected; loaded onto the column three times repeatedly; then, using 5 column volumes of water as the mobile phase, elute at a flow rate of 2 mL / min and discard the effluent; add 300 mL of 80 v / v% ethanol solution at a flow rate of 1 mL / min for elution, collect the effluent, determine the content, and then freeze-dry to obtain pure puerarin polyphenols (PLP).

[0039] The polyphenol content of each effluent was determined by the Folin-phenol method according to the standard curve equation, and the total phenol concentration in the desorption solution was measured. Calculate the adsorption capacity, adsorption rate, and desorption rate of the resin according to the formula.

[0040] Qe = (C0 - Ce)V / W Adsorption rate (%) = (C0 - Ce) × 100% Desorption rate (%) = C / (C0 - Ce) × 100% In the formula, Qe is the adsorption capacity, mg / g; C0 is the total phenol concentration of the added sample solution, mg / mL; Ce is the total phenol concentration of the unadsorbed part at adsorption equilibrium, mg / mL; V is the volume of the sample solution, mL; W is the mass of the resin, g; C is the total phenol concentration in the eluent, mg / mL.

[0041] 3. Determination of puerarin polyphenol concentration Standard curve: Take 0.1 mL, 0.3 mL, 0.5 mL, and 0.7 mL of gallic acid standard solution in sequence and place them in test tubes. Add 7.9 mL, 7.7 mL, 7.5 mL, and 7.3 mL of water respectively. Then add 0.5 mL of Folin-Ciocalteu reagent in sequence and mix well. Add 1.5 mL of 7.5 g / 100 mL sodium carbonate solution within 5 minutes and shake well. React in a water bath at 75 °C for 10 minutes and measure the absorbance A at 760 nm.

[0042] Use the gallic acid concentration as the abscissa and the absorbance value as the ordinate to plot the gallic acid standard curve.

[0043] Determination of polyphenol concentration: Take 1 mL of Pueraria lobata polyphenol sample solution in a test tube, add 7 mL of water and 0.5 mL of Folin reagent respectively, shake well, then add 1.5 mL of 7.5 g / 100 mL sodium carbonate solution and shake well. React in a water bath at 75 °C for 10 minutes and measure the absorbance A at 760 nm. Calculate the Pueraria lobata polyphenol concentration according to the standard curve.

[0044] Extract Pueraria lobata polyphenol by ultrasonic-assisted extraction, and the extraction rate is 5.89%; after purification by D-101 macroporous resin chromatography, the yield is 60.6%, the adsorption capacity of D-101 for Pueraria lobata polyphenol is 20.51 mg / g, the adsorption rate is 75.2%; the desorption rate is 85.1%.

[0045] 4. A composition with the effects of reducing uric acid and protecting the kidneys Mix the Pueraria lobata polyphenol obtained in step 1 and the polygonatum polysaccharide obtained in step 2 according to a mass ratio of 2:5 to obtain a composition (PPE) with the effects of reducing uric acid and protecting the kidneys.

[0046] Example 2: Preparation of a composition with the effects of reducing uric acid and protecting the kidneys Mix the Pueraria lobata polyphenol and polygonatum polysaccharide obtained in Example 1 of the present invention according to a mass ratio of 2:2 to obtain a composition (PPE) with the effects of reducing uric acid and protecting the kidneys.

[0047] Example 3: Preparation of a composition with the effects of reducing uric acid and protecting the kidneys Mix the Pueraria lobata polyphenol and polygonatum polysaccharide obtained in Example 1 of the present invention according to a mass ratio of 2:10 to obtain a composition (PPE) with the effects of reducing uric acid and protecting the kidneys.

[0048] Since Examples 1 to 3 have similar effects, for the convenience of subsequent discussion and reference, take the test results of Example 1 as an example. The following are the test results of Example 1.

[0049] The present invention constructs a hyperuricemia mouse model, and through drug administration treatment and index determination, explores the effects of the PPE composition on hyperuricemic mice, as follows.

[0050] 1. Experimental Methods 1. Mouse grouping, modeling and drug administration Thirty-two SPF male Kunming mice weighing 18g-22g were selected and allowed to drink water and eat freely. The temperature of the feeding room was about 25℃. After 7 days of adaptive feeding, they were randomly divided into 4 groups: normal control group (NC group), hyperuricemia mouse model group (MC group), positive control group (PC group), and polygonatum sibiricum polysaccharide and pueraria polyphenol combination group (PPE group). The hyperuricemia mouse model was established by intraperitoneal injection of potassium oxonate (300mg / kg / d.BW) and oral gavage of hypoxanthine (300mg / kg / d.BW) in the MC group, PC group and PPE group.

[0051] 1 hour after modeling, the NC group was gavaged with 0.5w / v% sodium carboxymethylcellulose (CMC-Na), the MC group was gavaged with 0.5w / v% CMC-Na, the PC group was gavaged with allopurinol (5mg / kg / d BW), and the PPE group was gavaged with the PPE composition prepared in Example 1 (350mg / kg / d), and the administration was continued for 14 days. 1 hour after the last day of administration, the mice were collected: the mice were anesthetized with ether, blood was collected from the orbits, and then the mice were immediately killed by cervical dislocation on an ice table, one kidney of the mouse was removed and stored in a -80°C refrigerator for testing, and the whole kidney tissue of the other side of the mouse was removed and fixed with 4% paraformaldehyde solution for renal HE staining pathological histological detection.

[0052] 2. Biochemical index detection The serum uric acid (UA), creatinine (CR) and urea (UREA) levels were measured by biochemical analyzer.

[0053] 2.1. ELISA was used to detect the expression levels of inflammatory factors such as TNF-α, IL-1β, and IL-6 in the kidney. The assay was performed according to the instructions of the kit.

[0054] 2.2. Pathological histological examination of mouse kidneys by HE staining. The samples were sent to Wuhan Saiweier Biotechnology Co., Ltd. for testing.

[0055] 2.3. WB was used to detect the expression level of mouse renal transporter 1 (URAT1).

[0056] Wash the tissue blocks three times with cold PBS to remove blood stains, dry them, and cut them into pieces with scissors. Mix the tissue pieces with RIPA solution at a ratio of 1 g:9 mL (protease inhibitors must be added to the RIPA solution before use), homogenize, let stand on ice for 30 minutes, centrifuge at 4°C, 12,000 rpm for 10 minutes, and collect the supernatant, which is the total protein solution.

[0057] Add the prepared samples to 5×SDS-PAGE Loading Buffer at a volume ratio of 1:4, boil and lyse the proteins for 10 min, incubate on ice for 10 min, then centrifuge at 4°C and 12,000 rpm for 10 min, and take the supernatant; Load the supernatant and perform SDS-PAGE gel electrophoresis; After electrophoresis, transfer the membrane and block it with 5% skim milk for 2 h; incubate overnight at 4°C with URAT1 primary antibody (1:3000) and β-Actin primary antibody (1:20,000). After incubation, wash the membrane 3 times with TBST, 8 min each time; then incubate with secondary antibody (1:10,000) for 2 h, and after incubation, wash the membrane 3 times with TBST, 8 min each time; finally, expose it with ECL color development solution, and analyze the gray value of the band after scanning using ImageJ software.

[0058] II. Experimental Results 1. Effects of PPE on blood uric acid, creatinine, and urea in hyperuricemia mice As Figure 1 shown, compared with the normal group, the serum uric acid and urea contents in the hyperuricemia model group mice were significantly increased ( p <0.01); the serum urea content in the positive control group mice was significantly increased ( p <0.05) and the creatinine content was also significantly increased ( p <0.01); there was no significant difference in the serum uric acid, creatinine, and urea contents between the PPE group mice and the normal group. Compared with the model group, the serum uric acid content in the positive group mice was extremely significantly decreased ( p <0.01); the serum uric acid level in the PPE group mice was extremely significantly decreased ( p <0.01) and the urea level was significantly decreased ( p <0.05).

[0059] 2. Effects of PPE on TNF-α, IL-1β, and IL-6 in the kidneys of hyperuricemia mice As Figure 2 shown, compared with the normal group, the IL-1β level in the kidneys of the model group and positive group mice was significantly increased ( p <0.05) and the TNF-α level was extremely significantly increased ( p <0.01), and the IL-6 level in the kidneys of the positive group mice was significantly increased ( p <0.05); compared with the model group, the TNF-α and IL-1β levels in the kidneys of the PPE group mice were both extremely significantly decreased ( p <0.01); there was no significant difference in the TNF-α, IL-1β, and IL-6 levels in the kidneys of the PPE group and the normal group.

[0060] 3. Histopathological examination of the kidneys AsFigure 3 As shown, the renal units of the mice in the normal group (NC) were intact in structure and normal in morphology. The glomeruli in the Bowman's capsule cavity were plump, and the renal tubule structure was normal. In the mice in the hyperuricemia model group (MC), the glomerular structure was loose, atrophied and fragmented. It could be seen that the renal tubular epithelial cells were swollen, necrotic, and severely vacuolated. The renal tubular lumen was dilated, and the internal brush border structure disappeared.

[0061] In the mice in the positive control group (PC), the glomeruli were atrophied, the renal tubular epithelial cells showed vacuolar degeneration, and the lumen was severely dilated. The internal brush border structure of the proximal convoluted tubule disappeared. There were a large number of interstitial spaces in the renal parenchyma, and brownish-black infiltrating inflammatory cells could be seen in the renal tubules and interstitium, indicating obvious kidney damage.

[0062] Compared with the model group and the positive control group, in the PPE group, the number of glomerular cells and the matrix were uniform. The dilation of the renal tubular lumen was significantly improved. The renal tubular epithelial cells were round and plump, the brush border was arranged neatly and regularly, and there was no obvious infiltration of inflammatory cells or other abnormalities. The renal structure and morphology were basically restored to normal. PPE had an obvious protective effect on kidney damage caused by hyperuricemia.

[0063] 4. Effect of PPE on the expression level of URAT1 protein in the kidneys of hyperuricemia mice The results were as Figure 4 shown. Compared with the normal group, the relative expression level of URAT1 protein in the kidneys of the mice in the model group was extremely significantly increased ( p <0.01); the relative expression levels of URAT1 protein in the kidneys of the mice in the positive control group and the PPE group were significantly lower than those in the model group ( p <0.05 or p <0.01), and there was no significant difference from the normal group.

[0064] Example 4: Preparation of a gel preparation with the effects of reducing uric acid and protecting the kidneys In this study, a total of 4 kinds of gel preparations were made, namely RPPE gel gummies, PPE gel gummies, RPPE gel jelly, and PPE gel jelly. The dosing amounts of each composition were calculated according to the drug dosage ratio of 9:1 for mice and humans.

[0065] The crude polygonatum polysaccharide obtained in Example 1 was labeled as RPCP, the polygonatum polysaccharide was labeled as PCP, the crude pueraria polyphenol was labeled as RPLP, and the pueraria polyphenol was labeled as PLP; the composition containing RPCP and RPLP was labeled as RPPE, and the composition containing PCP and PLP was labeled as PPE.

[0066] The preparation process of the 4 kinds of gel preparations was as follows: First, compound gel was made, and then the RPPE or PPE composition, auxiliary materials such as salt ions were dissolved in water → after mixing the compound gel and the dissolved RPPE, it was poured into a mold and left to stand until it cooled and formed.

[0067] 1. A kind of RPPE gel gummy candy is prepared by the following steps: (1) Prepare the compound gel: The ratio of the compound gelling agent is gelatin: carrageenan = 3.75 parts: 1.25 parts. Add water to 25 parts, and carry out a water bath at 60 °C, stir and mix evenly until fully dissolved.

[0068] (2) Dissolve RPPE: 1.55 parts of RPLP and 1.0 part of cyclodextrin. First, add 20 parts of water and stir to mix evenly, then add 3.85 parts of RPCP and mix well. Then add 2.5 parts of konjac gum and 0.1 part of KCl, and make up the water to 75 parts. Stir under the condition of a 100 °C water bath until fully dissolved.

[0069] (3) Mixing: Mix the compound gel in step (1) and the dissolved RPPE in step (2), stir and mix evenly, and when the temperature drops to 75 °C, pour into a mold and let it stand, and cool and form to obtain a kind of RPPE gel gummy candy, marked as preparation 1.

[0070] 2. A kind of RPPE gel gummy candy is prepared by the following steps: (1) Prepare the compound gel: The ratio of the compound gelling agent is gelatin: carrageenan = 3.75 parts: 1.25 parts. Add water to 25 parts, and carry out a water bath at 60 °C, stir and mix evenly until fully dissolved.

[0071] (2) Dissolve RPPE: 1.3 parts of RPLP and 1.0 part of cyclodextrin. First, add 20 parts of water and stir to mix evenly, then add 3.0 parts of RPCP and mix well. Then add 2.0 parts of konjac gum and 0.1 part of KCl, and make up the water to 75 parts. Stir under the condition of a 100 °C water bath until fully dissolved.

[0072] (3) Mixing: Mix the compound gel in step (1) and the dissolved RPPE in step (2), stir and mix evenly, and when the temperature drops to 75 °C, pour into a mold and let it stand, and cool and form to obtain a kind of RPPE gel gummy candy, marked as preparation 2.

[0073] 3. A kind of RPPE gel gummy candy is prepared by the following steps: (1) Prepare the compound gel: The ratio of the compound gelling agent is gelatin: carrageenan = 3.75 parts: 1.25 parts. Add water to 25 parts, and carry out a water bath at 60 °C, stir and mix evenly until fully dissolved.

[0074] (2) Dissolve RPPE: 2.5 parts of RPLP and 1.0 part of cyclodextrin. First, add 20 parts of water and stir to mix evenly, then add 5.0 parts of RPCP and mix well. Then add 4.0 parts of konjac gum and 0.1 part of KCl, and make up the water to 75 parts. Stir under the condition of a 100 °C water bath until fully dissolved.

[0075] (3) Mixing: Mix the compound gel from step (1) and the dissolved RPPE from step (2), stir well, and when cooled to 75 °C, pour into a mold and let stand. After cooling and forming, a kind of RPPE gel gummy is obtained, labeled as Preparation 3.

[0076] 4. A kind of PPE gel gummy is prepared by the following steps: (1) Preparing the compound gel: The ratio of the compound gelling agent is gelatin∶carrageenan = 3.75 parts∶1.25 parts. Add water to make 25 parts, and heat in a water bath at 60 °C, stir well and dissolve completely.

[0077] (2) Dissolving PPE: 1.4 parts of PLP, 1.0 part of cyclodextrin. First add 20 parts of water and stir well, then add 3.5 parts of PCP and mix well. 1 part of sodium carboxymethylcellulose, then add 2.5 parts of konjac gum and 0.1 part of KCl, and add water to make up to 75 parts. Stir under the condition of a 100 °C water bath until completely dissolved.

[0078] (3) Mixing: Mix the compound gel from step (1) and the dissolved PPE from step (2), stir well, and when cooled to 75 °C, pour into a mold and let stand. After cooling and forming, a kind of PPE gel gummy is obtained, labeled as Preparation 4.

[0079] 5. A kind of PPE gel gummy is prepared by the following steps: (1) Preparing the compound gel: The ratio of the compound gelling agent is gelatin∶carrageenan = 3.75 parts∶1.25 parts. Add water to make 25 parts, and heat in a water bath at 60 °C, stir well and dissolve completely.

[0080] (2) Dissolving PPE: 0.7 part of PLP, 1.0 part of cyclodextrin. First add 20 parts of water and stir well, then add 1.0 part of PCP and mix well. 1 part of sodium carboxymethylcellulose, then add 2.0 parts of konjac gum and 0.1 part of KCl, and add water to make up to 75 parts. Stir under the condition of a 100 °C water bath until completely dissolved.

[0081] (3) Mixing: Mix the compound gel from step (1) and the dissolved PPE from step (2), stir well, and when cooled to 75 °C, pour into a mold and let stand. After cooling and forming, a kind of PPE gel gummy is obtained, labeled as Preparation 5.

[0082] 6. A kind of PPE gel gummy is prepared by the following steps: (1) Preparing the compound gel: The ratio of the compound gelling agent is gelatin∶carrageenan = 3.75 parts∶1.25 parts. Add water to make 25 parts, and heat in a water bath at 60 °C, stir well and dissolve completely.

[0083] (2) Dissolution of PPE: 1.5 parts of PLP, 1.0 part of cyclodextrin. First, add 20 parts of water and stir to mix evenly, then add 2.0 parts of PCP and mix well. 1 part of sodium carboxymethylcellulose, then add 4.0 parts of konjac gum and 0.1 part of KCl. Add water to make up to 75 parts, and stir under the condition of a 100°C water bath until fully dissolved.

[0084] (3) Mixing: Mix the compound gel from step (1) and the dissolved PPE from step (2), stir and mix well, and when cooled to 75°C, pour into a mold and let stand, then cool and solidify to obtain a PPE gel gummy, labeled as Preparation 6.

[0085] 7. A RPPE gel jelly is prepared by the following steps: (1) Preparation of compound gel: The ratio of the compound gelling agent is gelatin∶carrageenan = 3.75 parts∶1.25 parts. Add water to 25 parts, and stir and mix well in a 60°C water bath until fully dissolved.

[0086] (2) Dissolution of RPPE: 1.55 parts of RPLP, 1.0 part of cyclodextrin. First, add 20 parts of water and stir to mix evenly, then add 3.85 parts of RPCP and mix well. Then add 0.75 part of pectin, add water, stir and mix well in a water bath. Finally, add 0.1 part of KCl and 0.1 part of CaCl₂, add water to make up to 75 parts, and stir under the condition of a 100°C water bath until fully dissolved.

[0087] (3) Mixing: Mix the compound gel from step (1) and the dissolved RPPE from step (2), stir and mix well, and when cooled to 75°C, pour into a mold and let stand, then cool and solidify to obtain a RPPE gel jelly, labeled as Preparation 7.

[0088] 8. A RPPE gel jelly is prepared by the following steps: (1) Preparation of compound gel: The ratio of the compound gelling agent is gelatin∶carrageenan = 3.75 parts∶1.25 parts. Add water to 25 parts, and stir and mix well in a 60°C water bath until fully dissolved.

[0089] (2) Dissolution of RPPE: 1.3 parts of RPLP, 1.0 part of cyclodextrin. First, add 20 parts of water and stir to mix evenly, then add 3.0 parts of RPCP and mix well. Then add 0.5 part of pectin, add water, stir and mix well in a water bath. Finally, add 0.1 part of KCl and 0.1 part of CaCl₂, add water to make up to 75 parts, and stir under the condition of a 100°C water bath until fully dissolved.

[0090] (3) Mixing: Mix the compound gel from step (1) and the dissolved RPPE from step (2), stir and mix well, and when cooled to 75°C, pour into a mold and let stand, then cool and solidify to obtain a RPPE gel jelly, labeled as Preparation 8.

[0091] 9. A RPPE gel jelly is prepared by the following steps: (1) Preparation of compound gel: The ratio of the compound gelling agent is gelatin∶carrageenan = 3.75 parts∶1.25 parts. Add water to make up 25 parts, and heat in a water bath at 60°C. Stir and mix well until fully dissolved.

[0092] (2) Dissolution of RPPE: 2.5 parts of RPLP and 1.0 part of cyclodextrin. First, add 20 parts of water and stir to mix well, then add 5.0 parts of RPCP and mix well, then add 2.0 parts of pectin, add water, stir and mix well in a water bath, and finally add 0.1 part of KCl and 0.1 part of CaCl₂. Add water to make up to 75 parts, and stir under the condition of a water bath at 100°C until fully dissolved.

[0093] (3) Mixing: Mix the compound gel in step (1) and the dissolved RPPE in step (2), stir and mix well, and when cooled to 75°C, pour into a mold and let stand, and cool to form a RPPE gel jelly, labeled as Preparation 9.

[0094] 10. A PPE gel jelly is prepared by the following steps: (1) Preparation of compound gel: The ratio of the compound gelling agent is gelatin∶carrageenan = 3.75 parts∶1.25 parts. Add water to make up 25 parts, and heat in a water bath at 60°C. Stir and mix well until fully dissolved.

[0095] (2) Dissolution of PPE: 1.4 parts of PLP and 1.0 part of cyclodextrin. First, add 20 parts of water and stir to mix well, then add 3.5 parts of PCP and mix well, 1 part of sodium carboxymethylcellulose, then add 0.75 part of pectin, add water, stir and mix well in a water bath, and finally add 0.1 part of KCl and 0.1 part of CaCl₂. Add water to make up to 75 parts, and stir under the condition of a water bath at 100°C until fully dissolved.

[0096] (3) Mixing: Mix the compound gel in step (1) and the dissolved PPE in step (2), stir and mix well, and when cooled to 75°C, pour into a mold and let stand, and cool to form a PPE gel jelly, labeled as Preparation 10.

[0097] 11. A PPE gel jelly is prepared by the following steps: (1) Preparation of compound gel: The ratio of the compound gelling agent is gelatin∶carrageenan = 3.75 parts∶1.25 parts. Add water to make up 25 parts, and heat in a water bath at 60°C. Stir and mix well until fully dissolved.

[0098] (2) Dissolution of PPE: 0.7 part of PLP and 1.0 part of cyclodextrin. First, add 20 parts of water and stir to mix well, then add 1.0 part of PCP and mix well, 1 part of sodium carboxymethylcellulose, then add 0.5 part of pectin, add water, stir and mix well in a water bath, and finally add 0.1 part of KCl and 0.1 part of CaCl₂. Add water to make up to 75 parts, and stir under the condition of a water bath at 100°C until fully dissolved.

[0099] (3) Mixing: Mix the compound gel from step (1) and the dissolved PPE from step (2), stir well, and when cooled to 75°C, pour into a mold and let stand. After cooling and forming, a PPE gel jelly is obtained, labeled as Preparation 11.

[0100] 12. A PPE gel jelly is prepared by the following steps: (1) Preparing the compound gel: The ratio of the compound gelling agent is gelatin∶carrageenan = 3.75 parts∶1.25 parts. Add water to 25 parts, and perform a water bath at 60°C, stir well, and dissolve completely.

[0101] (2) Dissolving PPE: 1.5 parts of PLP, 1.0 part of cyclodextrin. First, add 20 parts of water and stir well, then add 2.0 parts of PCP and mix well, 1 part of sodium carboxymethylcellulose, then add 2.0 parts of pectin, add water, perform a water bath and stir well. Finally, add 0.1 part of KCl and 0.1 part of CaCl₂, and make up the water to 75 parts. Stir under the condition of a 100°C water bath until completely dissolved.

[0102] (3) Mixing: Mix the compound gel from step (1) and the dissolved PPE from step (2), stir well, and when cooled to 75°C, pour into a mold and let stand. After cooling and forming, a PPE gel jelly is obtained, labeled as Preparation 12.

[0103] Since Preparations 1 - 3 have similar effects, Preparations 4 - 6 have similar effects, Preparations 7 - 9 have similar effects, and Preparations 10 - 12 have similar effects, for the convenience of subsequent discussion and reference, the test results of Preparations 1, 4, 7, and 10 are taken as examples. The following are the test results of Preparations 1, 4, 7, and 10.

[0104] Using the TA.GEL texture analyzer of Shanghai Baosheng Industrial Development Co., Ltd., with the probe model TA / 0.5, the hardness, viscosity, elasticity, chewiness, adhesiveness, cohesiveness, and resilience of Preparations 1, 4, 7, and 10 are detected respectively.

[0105] Detection parameters of RPPE gel gummies and PPE gel gummies: Pre-test speed 2.0 mm / s, test speed 1.0 mm / s, post-test speed 1 mm / s, contact force 5.0 gf, compression variable 50%.

[0106] The texture parameters of RPPE gel gummies and PPE gel gummies are shown in Table 1: Table 1 Texture parameters of RPPE gel gummies and PPE gel gummies Detection parameters of RPPE gel jelly and PPE gel jelly: speed before testing is 2.0 mm / s, testing speed is 1.0 mm / s, speed after testing is 1 mm / s, contact force is 5.0 gf, and compression variable is 30%.

[0107] The texture parameters of RPPE gel jelly and PPE gel jelly are shown in Table 2: Table 2 Texture parameters of RPPE gel jelly and PPE gel jelly Tables 1 to 2 and Figures 5 - 6 The comparison results show that both the hardness and viscosity of RPPE gel jelly and PPE gel jelly are relatively low. Therefore, gel jelly is more suitable for people with poor teeth. While the overall effect of RPPE gel candy and PPE gel candy is better, with moderate hardness and appropriate elasticity, adhesiveness, chewiness and gumminess. The overall chewiness is suitable for most people.

[0108] In modern society where health is increasingly emphasized, functional leisure foods have gradually attracted people's attention and research. Functional gel candies or gel jellies with high moisture, low sweetness and low calories have received more and more attention. Both PCP and PLP are natural bioactive substances extracted from medicinal and edible homologous herbs. As the main components of the gel composition and its efficacy, they are not only safe and non-toxic, but also the various bioactivities such as reducing uric acid they possess are of great benefit to people's health.

[0109] It should be noted that when the present invention involves numerical ranges, it should be understood that any value between the two endpoints of each numerical range and the two endpoints can be selected. Since the adopted step methods are the same as those in the embodiments, in order to prevent repetition, the present invention describes the preferred embodiments. Although the preferred embodiments of the present invention have been described, those skilled in the art can make additional changes and modifications to these embodiments once they know the basic creative concept. Therefore, the appended claims are intended to be interpreted to include the preferred embodiments and all changes and modifications falling within the scope of the present invention.

[0110] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalent technologies, the present invention is also intended to include these modifications and variations.

Claims

1. A composition with the effects of reducing uric acid and protecting the kidneys, characterized in that The composition is prepared by mixing pueraria polyphenols and polygonatum polysaccharides in a mass ratio of 2:2 to 10.

2. The composition according to claim 1, characterized in that, The pueraria polyphenols are obtained by extracting crude polyphenols from pueraria by alcohol extraction method, purifying by column chromatography, and concentrating.

3. The composition according to claim 1, characterized in that, The polygonatum polysaccharides are obtained by extracting crude polysaccharides from polygonatum cyrtonema hua by ultrasonic-assisted water extraction method and purifying by deproteinization.

4. Use of the composition according to any one of claims 1 to 3 in the preparation of a drug for treating hyperuricemia.

5. Use of the composition according to claim 4 in the preparation of a medicament for treating hyperuricemia, characterized in that, The drug for treating hyperuricemia is composed of the composition and excipients.

6. Use of the composition according to claim 4 in the preparation of a medicament for treating hyperuricemia, characterized in that, The dosage form of the drug for treating hyperuricemia is dispersible tablets, soft capsule agents, granules, powders, dripping pills, gels or oral liquid preparations.

7. Use of the composition according to claim 4 in the preparation of a medicament for treating hyperuricemia, characterized in that, The drug for treating hyperuricemia is a drug for reducing uric acid content, reducing urea content or alleviating renal pathological damage.

8. Use of the composition according to any one of claims 1 to 3 in the preparation of a functional food, characterized in that, The functional food is a jelly-type gel or a gummy gel.

9. The application according to claim 8, characterized in that, The gummy gel is composed of, by mass: 0.7 parts to 1.5 parts of pueraria polyphenols, 1.0 parts to 3.5 parts of polygonatum polysaccharides, 3.75 parts of gelatin, 1.25 parts of carrageenan, 1.0 part of cyclodextrin, 1 part of sodium carboxymethylcellulose, 2 parts to 4 parts of konjac gum, and 0.1 part of KCl.

10. The application according to claim 8, wherein The jelly gel is composed of, by mass: 0.7 parts to 1.5 parts of pueraria polyphenols, 1.0 parts to 3.5 parts of polygonatum polysaccharides, 3.75 parts of gelatin, 1.25 parts of carrageenan, 1.0 part of cyclodextrin, 1 part of sodium carboxymethylcellulose, 0.5 parts to 2 parts of pectin, 0.1 part of KCl, and 0.1 part of CaCl2.

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