Composition containing rhizoma dioscoreae nipponicae and application of composition in preparation of medicine for treating hyperuricemia

Abstract

The invention belongs to the technical field of medicines for treating hyperuricemia, and particularly relates to application of a composition containing rhizoma dioscoreae nipponicae to preparation of medicines for treating hyperuricemia. The composition containing the rhizoma dioscoreae nipponicae is prepared from the rhizoma dioscoreae nipponicae, sodium chloride and zinc sulfate, and the mass ratio of the rhizoma dioscoreae nipponicae to the sodium chloride to the zinc sulfate is 1000: (1-2): (0-1). The traditional Chinese medicine composition has the effects of reducing uric acid, urea nitrogen and the like, and can treat hyperuricemia. The preparation process is crushing and mixing, does not need complex extraction processes such as heating reflux and ultrasonic extraction, does not need extraction solvents such as ethanol, does not have reagent residues, does not pollute the environment, and is energy-saving and environment-friendly.

Description

Technical Field

[0001] This invention belongs to the technical field of drugs for the treatment of hyperuricemia, specifically relating to the application of a composition containing *Dioscorea nipponica* in the preparation of drugs for the treatment of hyperuricemia. Background Technology

[0002] Hyperuricemia is a metabolic disorder caused by purine metabolism disorders and / or decreased uric acid excretion, characterized by elevated serum uric acid levels. Uric acid is produced by purine metabolism, with about one-third originating from diet and the remainder from endogenous sources. Causes of hyperuricemia include: 1) excessive uric acid production, such as a high-purine diet or congenital enzyme deficiencies; 2) decreased excretion, such as renal insufficiency or drug effects. Hyperuricemia can be primary or secondary. Most patients are asymptomatic, but long-term hyperuricemia can lead to gout, gouty nephropathy, and uric acid stones. Furthermore, hyperuricemia is an independent risk factor for cardiovascular disease, chronic kidney disease, and metabolic syndrome, and is associated with increased all-cause mortality. Therefore, the development of therapeutic drugs for hyperuricemia is essential.

[0003] Although existing research shows that total saponins from *Dioscorea nipponica* can regulate the intestinal flora and short-chain fatty acids in hyperuricemic rats—for example, total saponins from *Dioscorea nipponica* can significantly increase the abundance of Firmicutes, Lactobacillus, Clostridium, Rumenobacter, Lactobacillus rhamnosus, Lactobacillus johnsonii, and Lactobacillus reuteri in the intestinal flora of hyperuricemic rats—and that total saponins from *Dioscorea nipponica* have a good uric acid-lowering ability, they also demonstrate this effect.

[0004] However, total saponins from Dioscorea nipponica are active substances that need to be extracted from Dioscorea nipponica. Due to the complex extraction and purification process, the preparation process is troublesome. There is a need to provide a hyperuricemia treatment product with a simple preparation process. Summary of the Invention

[0005] To address the aforementioned technical problems, this invention provides an application of a composition containing *Dioscorea nipponica* in the preparation of a drug for treating hyperuricemia.

[0006] The purpose of this invention is to provide a composition containing *Dioscorea nipponica* for treating hyperuricemia, which is prepared from *Dioscorea nipponica*, sodium chloride and zinc sulfate, wherein the mass ratio of *Dioscorea nipponica*, sodium chloride and zinc sulfate is 1000:1~2:0~1.

[0007] Dioscorea nipponica is rich in saponins, which show promise in treating hyperuricemia. However, besides saponins, it also contains other components. When comparing equal masses of Dioscorea nipponica and its total saponins, the total saponins from the raw Dioscorea nipponica are present in lower quantities than when using it directly. Therefore, the experimental results of this invention show that using Dioscorea nipponica alone as a medicinal agent is less effective in lowering uric acid and urea nitrogen than using total saponins alone.

[0008] To overcome this deficiency, this invention combines *Dioscorea nipponica* with sodium chloride and zinc sulfate. Sodium chloride and zinc sulfate, on the one hand, regulate the body's electrolyte balance, and on the other hand, enhance the effects of *Dioscorea nipponica* in lowering uric acid and urea nitrogen to varying degrees. The optimal effect in lowering uric acid and urea nitrogen is achieved when all three are used together.

[0009] In addition, sodium ions can regulate fluid balance and help maintain normal blood pressure. Zinc ions have anti-inflammatory and antioxidant properties, and can also participate in various immune functions, enhancing the body's immune response and helping to alleviate metabolic disorders caused by hyperuricemia.

[0010] Preferably, in the above-mentioned composition containing *Dioscorea nipponica* for treating hyperuricemia, the mass ratio of *Dioscorea nipponica*, sodium chloride, and zinc sulfate is 1000:1.5:0.5.

[0011] Preferably, the above-mentioned composition containing *Dioscorea nipponica* for treating hyperuricemia is prepared according to the following method: Dry the pangolin and grind it into powder; Prepare the powdered *Dioscorea nipponica*, sodium chloride, and zinc sulfate according to the mass ratio; Mix thoroughly to obtain a composition containing *Dioscorea nipponica* for the treatment of hyperuricemia.

[0012] Preferably, in the above-mentioned composition containing *Dioscorea nipponica* for treating hyperuricemia, the powder of *Dioscorea nipponica* is 100-200 mesh.

[0013] Preferably, in the application of the above-mentioned composition containing *Dioscorea nipponica* in the preparation of a drug for treating hyperuricemia, the composition containing *Dioscorea nipponica* for treating hyperuricemia is used as the sole active ingredient, combined with pharmaceutically acceptable excipients, to prepare a drug for treating hyperuricemia.

[0014] Preferably, in the application of the above-mentioned composition containing *Dioscorea nipponica* in the preparation of a drug for treating hyperuricemia, the pharmaceutically acceptable excipients include lubricants, adhesives, or surfactants.

[0015] Preferably, in the application of the above-mentioned composition containing *Dioscorea nipponica* in the preparation of a drug for treating hyperuricemia, the lubricant is methyl stearate. The adhesive is polyvinyl ketone; The surfactant is sodium dodecyl sulfate or polyoxyethylene fatty acid ester.

[0016] Preferably, the above-mentioned composition containing *Dioscorea nipponica* is used in the preparation of a drug for treating hyperuricemia, wherein the drug for treating hyperuricemia refers to a drug that reduces the uric acid content and urinary nitrogen content in the body.

[0017] Preferably, in the application of the above-mentioned composition containing *Dioscorea nipponica* in the preparation of a drug for treating hyperuricemia, the uric acid content refers to its content in serum; The urinary nitrogen content refers to its concentration in serum.

[0018] Preferably, the above-mentioned composition containing *Dioscorea nipponica* is used in the preparation of a drug for treating hyperuricemia, wherein the drug for treating hyperuricemia refers to a drug that inhibits the growth of intestinal pathogens.

[0019] Compared with the prior art, the present invention has the following beneficial effects: One of the innovative aspects of this invention is the combination of *Dioscorea nipponica* with sodium chloride and zinc sulfate in a mass ratio of 1000:1~2:0~1. Sodium chloride and / or zinc sulfate enhance the effects of *Dioscorea nipponica* in lowering uric acid and urea nitrogen levels. This can treat hyperuricemia, especially showing excellent effects in lowering uric acid and urea nitrogen levels in hyperuricemia model animals. Preferably, *Dioscorea nipponica* is prepared as a powder.

[0020] The second innovation of this invention lies in the fact that the prices of Dioscorea nipponica, sodium chloride, and zinc sulfate are much lower than the price of total saponins from Dioscorea nipponica, thus reducing the cost of preparing the composition and helping to alleviate the medical burden on patients, which has great application and promotion value.

[0021] The third innovation of this invention is that the main preparation process of the composition provided by this invention is pulverization and mixing, which does not require complex extraction processes such as heating reflux and ultrasonic extraction, nor does it require extraction solvents such as ethanol. There are no reagent residues, no environmental pollution, and it is energy-saving and environmentally friendly.

[0022] The fourth innovation of this invention is that the composition provided by this invention can selectively inhibit bacteria, and has no significant inhibitory effect on Bacillus subtilis, a beneficial intestinal bacterium, but has a significant inhibitory effect on intestinal pathogens. Attached Figure Description

[0023] Figure 1 This is a technology roadmap. Detailed Implementation

[0024] To enable those skilled in the art to better understand and implement the technical solutions of the present invention, the present invention will be further described below in conjunction with specific embodiments and accompanying drawings.

[0025] Unless otherwise specified, all reagents used in this invention are commercially available, and all methods used are conventional techniques in the art. Figure 1 This is the technical approach of the present invention.

[0026] Example 1 A composition containing *Dioscorea nipponica* for treating hyperuricemia, wherein the composition is prepared from *Dioscorea nipponica* and sodium chloride in a mass ratio of 1000:1.

[0027] It is prepared according to the following method: Dry the pangolin and pulverize it into 100-mesh powder; Prepare the powdered *Dioscorea nipponica* and sodium chloride at a mass ratio of 1000:1; Mix thoroughly to obtain a composition containing *Dioscorea nipponica* for the treatment of hyperuricemia, hereinafter referred to as drug 1.

[0028] Example 2 A composition containing *Dioscorea nipponica* for treating hyperuricemia, wherein the composition is prepared from *Dioscorea nipponica* and sodium chloride in a mass ratio of 1000:2.

[0029] It is prepared according to the following method: Dry the pangolin and pulverize it into 100-mesh powder; Prepare the powdered *Dioscorea nipponica* and sodium chloride at a mass ratio of 1000:2; Mix thoroughly to obtain a composition containing *Dioscorea nipponica* for the treatment of hyperuricemia, referred to as drug 2 in the following experiment.

[0030] Example 3 A composition containing *Dioscorea nipponica* for treating hyperuricemia, wherein the composition is prepared from *Dioscorea nipponica* and sodium chloride in a mass ratio of 1000:1.

[0031] It is prepared according to the following method: Dry the pangolin and pulverize it into 200-mesh powder; Prepare the powdered *Dioscorea nipponica* and sodium chloride at a mass ratio of 1000:1; Mix thoroughly to obtain a composition containing *Dioscorea nipponica* for the treatment of hyperuricemia, hereinafter referred to as drug 3 in this experiment.

[0032] Example 4 A composition containing *Dioscorea nipponica* for treating hyperuricemia, said composition being prepared from *Dioscorea nipponica*, sodium chloride, and zinc sulfate in a mass ratio of 1000:1:0.5.

[0033] It is prepared according to the following method: Dry the pangolin and pulverize it into 100-mesh powder; Prepare the powdered smilax, sodium chloride, and zinc sulfate in a mass ratio of 1000:1:0.5; Mix thoroughly to obtain a composition containing *Dioscorea nipponica* for the treatment of hyperuricemia, referred to as drug 4 in the following experiment.

[0034] Example 5 A composition containing *Dioscorea nipponica* for treating hyperuricemia, said composition being prepared from *Dioscorea nipponica*, sodium chloride, and zinc sulfate in a mass ratio of 1000:1:1.

[0035] It is prepared according to the following method: Dry the pangolin and pulverize it into 100-mesh powder; Prepare the powdered smilax, sodium chloride, and zinc sulfate in a mass ratio of 1000:1:0.5; Mix thoroughly to obtain a composition containing *Dioscorea nipponica* for the treatment of hyperuricemia, referred to as drug 5 in the following experiment.

[0036] Control group 1 A pharmaceutical preparation consisting only of 100-mesh dried pangolin powder. This is a single-factor control.

[0037] Control group 2 A pharmaceutical preparation, namely total saponins from *Dioscorea nipponica*. The saponins are prepared as follows: 1 g of *Dioscorea nipponica* is pulverized into a 60-mesh fine powder, and 20 mL of a 30% (v / v) ethanol solution is added. The mixture is then ultrasonically extracted for 30 min at a power of 200 W and a frequency of 40 kHz. After ultrasonic extraction, the mixture is centrifuged at 1000 r / min for 15 min. The supernatant is collected, filtered through rapid qualitative filter paper, and evaporated to dryness. The residue is dissolved in water and extracted twice with 15 mL of ether each time. The aqueous layers are combined, and the mixture is extracted twice with 20 mL of water-saturated n-butanol each time. The n-butanol layer is collected and freeze-dried to obtain a powder. The crude saponins are obtained as a solid, and the crude saponins used in the following experiments are the crude saponins.

[0038] Control group 2 was a positive control for total saponins from Dioscorea nipponica.

[0039] Control group 3 The drug used was sodium chloride alone. Since excessive intake of inorganic salts can cause metabolic disorders in rats, and it is impossible for humans to directly ingest excessive amounts of inorganic salts to administer the drug, the dosage of sodium chloride used in this control group was the same as that used in Example 4. This is a single-factor control.

[0040] Control group 4 The drug used was zinc sulfate alone. Since excessive intake of inorganic salts can cause metabolic disorders in rats, the *Dioscorea nipponica* and sodium chloride were replaced with an equal volume of sterile water. Furthermore, it is impossible for humans to directly ingest excessive amounts of inorganic salts when taking medication; therefore, the dosage of zinc sulfate used in this control group was the same as that used in Example 4. This is a single-factor control.

[0041] Control group 5 (The enhancing effect of zinc sulfate was slightly weaker, so it is recommended to combine it with sodium chloride). A pharmaceutical preparation comprising a composition containing *Dioscorea nipponica* prepared from *Dioscorea nipponica* and zinc sulfate in a mass ratio of 1000:0.5. This is a two-factor control.

[0042] It is prepared according to the following method: Dry the pangolin and pulverize it into 100-mesh powder; Prepare the powdered smilax and zinc sulfate at a mass ratio of 1000:0.5; The mixture was thoroughly mixed to obtain a composition containing *Dioscorea nipponica* for the treatment of hyperuricemia. This composition will be referred to as drug 6 in the following experiments.

[0043] Experiment 1: An Experiment on the Treatment of Hyperuricemia (1) Rats SPF-grade male SD rats, 8 weeks old, weighing 200±15g, were purchased from the Experimental Animal Center of Heilongjiang University of Traditional Chinese Medicine.

[0044] (2) Adaptive feeding All rats were acclimatized for 7 days under an ambient temperature of 24℃, relative humidity of 55%, natural light, and free access to water and food.

[0045] (3) Grouping The study included a blank control group, a model group, Example 1 group, Example 2 group, Example 3 group, Example 4 group, Example 5 group, and control group 1, control group 2, control group 3, control group 4, and control group 5. Each group contained 10 rats.

[0046] (4) Processing Control group: From day 1 to day 35 after the end of the acclimatization period, at 9:00 am every day, 250 mg / kg body weight (calculated after weighing body weight weekly) of sterile water was administered by gavage, and normal feed was given.

[0047] From day 15 to 35, administer 400 mg / kg body weight (calculated after weekly body weight measurement) of sterile water via gavage at 5 PM daily.

[0048] Model group: From day 1 to day 35 after the end of the acclimatization period, the patient was fed potassium oxonate solution (5 g potassium oxonate + 95 mL physiological saline + 0.5 g sodium carboxymethyl cellulose) at a dose of 250 mg / kg body weight (calculated after weighing body weight weekly). At the same time, the patient was fed 10% yeast feed (10 wt% yeast added to the basal feed).

[0049] From day 15 to 35, administer 400 mg / kg body weight (calculated after weekly body weight measurement) of sterile water via gavage at 5 PM daily.

[0050] Example 1 Group: From day 1 to day 35 after the end of the acclimatization period, the patient was fed potassium oxonate solution (5 g potassium oxonate + 95 mL physiological saline + 0.5 g sodium carboxymethyl cellulose) at a dose of 250 mg / kg body weight (calculated after weighing body weight weekly). At the same time, the patient was fed 10% yeast feed (10 wt% yeast added to the basic feed).

[0051] From day 15 to 35, at 5 p.m. each day, 0.5 g of drug 1 from Example 1 was taken and suspended in 100 mL of sterile water to obtain suspension 1, which was then administered by gavage. The dosage of drug 1 was the same as that of drug 4, based on the dosage of *Dioscorea nipponica*.

[0052] Example 2 Group: From day 1 to day 35 after the end of the acclimatization period, the patient was fed potassium oxonate solution (5 g potassium oxonate + 95 mL physiological saline + 0.5 g sodium carboxymethyl cellulose) at a dose of 250 mg / kg body weight (calculated after weighing body weight weekly). At the same time, the patient was fed 10% yeast feed (10 wt% yeast added to the basic feed).

[0053] From day 15 to 35, at 5 p.m. each day, 0.5 g of drug 2 from Example 2 was taken and suspended in 100 mL of sterile water to obtain suspension 2. This suspension was administered by gavage. The dosage of drug 2 was the same as that of drug 4, based on the dosage of *Dioscorea nipponica*.

[0054] Example 3 Group: From day 1 to day 35 after the end of the acclimatization period, the patient was fed potassium oxonate solution (5 g potassium oxonate + 95 mL physiological saline + 0.5 g sodium carboxymethyl cellulose) at a dose of 250 mg / kg body weight (calculated after weighing body weight weekly). At the same time, the patient was fed 10% yeast feed (10 wt% yeast added to the basic feed).

[0055] From day 15 to 35, at 5 p.m. each day, 0.5 g of drug 3 from Example 3 was taken and suspended in 100 mL of sterile water to obtain suspension 3. This suspension was then administered by gavage. The dosage of drug 3 was the same as that of drug 4, based on the dosage of *Dioscorea nipponica*.

[0056] Example 4 Group: From day 1 to day 35 after the end of the acclimatization period, the patient was fed potassium oxonate solution (5 g potassium oxonate + 95 mL physiological saline + 0.5 g sodium carboxymethyl cellulose) at a dose of 250 mg / kg body weight (calculated after weighing body weight weekly). At the same time, the patient was fed 10% yeast feed (10 wt% yeast added to the basic feed).

[0057] From day 15 to 35, at 5 p.m. each day, 0.5 g of drug 4 from Example 4 was taken and suspended in 100 mL of sterile water to obtain suspension 4, which was then administered by gavage. The dosage of suspension 4 of drug 4 was 400 mg / kg body weight (calculated after weighing body weight weekly).

[0058] Example 5 group: From day 1 to day 35 after the end of the acclimatization period, the patient was fed potassium oxonate solution (5 g potassium oxonate + 95 mL physiological saline + 0.5 g sodium carboxymethyl cellulose) at a dose of 250 mg / kg body weight (calculated after weighing body weight weekly). At the same time, the patient was fed 10% yeast feed (10 wt% yeast added to the basic feed).

[0059] From day 15 to 35, at 5 p.m. each day, 0.5 g of drug 5 from Example 5 was taken and suspended in 100 mL of sterile water to obtain suspension 5, which was then administered by gavage. The dosage of drug 5 was the same as that of drug 4, based on the dosage of *Dioscorea nipponica*.

[0060] Control Group 1: From day 1 to day 35 after the end of the acclimatization period, the patient was fed potassium oxonate solution (5 g potassium oxonate + 95 mL physiological saline + 0.5 g sodium carboxymethyl cellulose) at a dose of 250 mg / kg body weight (calculated after weighing body weight weekly). At the same time, the patient was fed 10% yeast feed (10 wt% yeast added to the basal feed).

[0061] From day 15 to day 35, at 5 p.m. each day, 0.5 g of the drug from control group 1 was suspended in 100 mL of sterile water and administered by gavage. The dosage of control group 1 was the same as that of drug 4, based on the dosage of *Dioscorea nipponica*.

[0062] Control group 2: From day 1 to day 35 after the end of the acclimatization period, the patients were fed potassium oxonate solution (5 g potassium oxonate + 95 mL physiological saline + 0.5 g sodium carboxymethyl cellulose) at a dose of 250 mg / kg body weight (calculated after weighing body weight weekly) and 10% yeast feed (10 wt% yeast added to the basal feed).

[0063] From day 15 to 35, at 5 p.m. each day, 0.5 g of the drug from control group 2 was suspended in 100 mL of sterile water and administered by gavage. The dosage of total saponins from *Dioscorea nipponica* in control group 2 was the same as the dosage of *Dioscorea nipponica* in drug 4.

[0064] Control group 3: From day 1 to day 35 after the end of the acclimatization period, the patients were fed potassium oxonate solution (5 g potassium oxonate + 95 mL physiological saline + 0.5 g sodium carboxymethyl cellulose) at a dose of 250 mg / kg body weight (calculated after weighing body weight weekly) and 10% yeast feed (10 wt% yeast added to the basal feed).

[0065] From day 15 to day 35, at 5 p.m. each day, 0.5 g of the drug from control group 3 was taken, suspended in 100 mL of sterile water, and administered by gavage. The dosage of the drug in control group 3 was the same as that in Example 4, calculated as sodium chloride.

[0066] Control group 4: From day 1 to day 35 after the end of the acclimatization period, the patients were fed potassium oxonate solution (5 g potassium oxonate + 95 mL physiological saline + 0.5 g sodium carboxymethyl cellulose) at a dose of 250 mg / kg body weight (calculated after weighing body weight weekly) and 10% yeast feed (10 wt% yeast added to the basal feed).

[0067] From day 15 to 35, at 5 p.m. each day, 0.5 g of the drug from control group 4 was suspended in 100 mL of sterile water and administered by gavage. The dosage of the drug in control group 4 was the same as that in example 4, calculated as zinc sulfate.

[0068] Control group 5: From day 1 to day 35 after the end of the acclimatization period, the patients were fed potassium oxonate solution (5 g potassium oxonate + 95 mL physiological saline + 0.5 g sodium carboxymethyl cellulose) at a dose of 250 mg / kg body weight (calculated after weighing body weight weekly) and 10% yeast feed (10 wt% yeast added to the basal feed).

[0069] From day 15 to day 35, at 5 p.m. each day, 0.5 g of the drug from control group 5 was suspended in 100 mL of sterile water and administered by gavage. The dosage of the drug from control group 5 was the same as that of drug 4, based on the dosage of *Dioscorea nipponica*.

[0070] It should be noted that sodium chloride and zinc sulfate are only used as synergistic enhancers and are not used alone. Control groups 3 and 4 are to demonstrate that using the inorganic salts in the dosage of Example 4 alone is not effective in reducing serum uric acid and serum urea nitrogen. Therefore, the above experimental groups only used sodium chloride or zinc sulfate in the same dosage as the inorganic salts in Example 4. Since the proportion of sodium chloride and zinc sulfate in the composition is only a few parts per thousand of the *Dioscorea nipponica*, if the above experiments directly used the same dosage of inorganic salts as all the components of the drug in Example 4, the dosage would be too large and may affect other physiological metabolisms in rats, leading to distorted results.

[0071] (5) Measurement of serum uric acid and blood urea nitrogen Serum samples were collected on days 14, 28, and 35, and the levels of serum uric acid and blood urea nitrogen were measured using a fully automated biochemical analyzer. Serum uric acid results are shown in Table 1. Serum urea nitrogen results are shown in Table 2.

[0072] Table 1. Serum uric acid test results (mmol / L) Table 2. Serum urea nitrogen test results (mmol / L) Serum uric acid is a direct indicator for evaluating hyperuricemia and is clinically used for diagnosis, risk assessment, and guiding drug initiation and dosage adjustment. Serum urea nitrogen is an indirect indicator of impaired renal function or excessive protein breakdown; in the assessment of hyperuricemia, it can serve as a background value for renal function or an indication of complications. Therefore, testing these two indicators is of great significance for evaluating the efficacy of drug treatment for hyperuricemia.

[0073] As can be seen from the results in Tables 1 and 2, after 35 days, each example group was able to adjust serum uric acid and serum urea nitrogen to levels close to the blank group, suggesting that it has the effect of improving and treating hyperuricemia. Control group 1, which was simply *Dioscorea nipponica* powder, showed a significantly lower therapeutic effect than the example groups. Control group 2 was a positive control of total saponins from *Dioscorea nipponica*, which, although having a significant effect in lowering uric acid and urea nitrogen, requires a complex extraction process and is costly. Compared with this, the preparation process of the agent in the embodiments of the present invention is simple, the cost is low, and the therapeutic effect is basically equivalent. From a cost perspective, the agent in the embodiments of the present invention is the preferred option. Control group 3 was sodium chloride, and control group 4 was zinc sulfate, neither of which showed significant therapeutic effects. Control group 5 was a combination of *Dioscorea nipponica* and zinc sulfate, but its therapeutic effect was slightly worse than that of the example groups. By comparing the various example groups with control group 1 and control group 5, it was found that when the powder of *Dioscorea nipponica* is combined with sodium chloride and / or zinc sulfate, the effect of reducing uric acid and urea nitrogen is much higher than that of the single-factor group of *Dioscorea nipponica* powder (control group 1). This indicates that sodium chloride and / or zinc sulfate enhance the effect of *Dioscorea nipponica* in reducing uric acid and urea nitrogen.

[0074] Experiment 2: Experiment on selective inhibition of intestinal bacteria (1) Preparation of bacterial suspension Materials preparation: Prepare freshly cultured Clostridium difficile standard strain BNCC364175, Salmonella enterica serovar Typhimurium standard strain, and Bacillus subtilis BNCC189983.

[0075] Prepare sterile water.

[0076] After activation, the concentration of the bacterial suspensions of each of the three strains was adjusted to 1.0 × 10⁻⁶ using sterile water. 5 cfu / mL.

[0077] (2) Operating environment: The procedure is performed in a laminar flow hood, which provides a sterile environment. Detection process: The experimental group used a concentration of 1.0 × 10⁻⁶. 5 1 mL of a bacterial suspension with a concentration of CFU / mL was added to 5 mL of the test liquid, which was any of the reagents used in the examples and control group. The mixture was incubated at room temperature (25°C) for 20 min. Simultaneously, a 1.0 × 10⁻⁶ CFU / mL solution was prepared. 5The bacterial suspension at CFU / mL served as the blank control group; all other procedures were the same as in the experimental group. The viable bacterial counts for each group were recorded after treatment.

[0078] In the test liquid, the concentrations of Examples 1-5, Control Group 1, and Control Group 5 were 1 g / 100 mL, based on 1 g of *Dioscorea nipponica* mass; the concentration of Control Group 3 was 1 g / 100 mL, the concentration of Control Group 4 was 0.001 g / 100 mL, and the concentration of Control Group 5 was 0.0005 g / 100 mL. Since the proportion of inorganic salts was only a few parts per thousand of *Dioscorea nipponica*, the resulting volume changes and their impact on viable bacterial concentration were ignored.

[0079] The antibacterial rate can be calculated using the following formula.

[0080] 00%.

[0081] The results of the antibacterial test are shown in Table 1.

[0082] Table 3. Statistical results of antibacterial rate The research results of "Liu Shumin, Lu Yi, Xuan Xiuxiu, et al. Investigation on the effects and mechanisms of total saponins from Dioscorea nipponica on hyperuricemic rats based on 16S rRNA sequencing [J]. Chinese Journal of Medicine, 2024, 000(1):5" showed that there were significant differences in the abundance of intestinal microorganisms between rats with hyperuricemia and rats with low uric acid levels. For example, saponins from Dioscorea nipponica can reduce urinary nitrogen levels, regulate the abundance of Proteobacteria, decrease the abundance of Bacteroidetes, and increase the abundance of Lactobacillus, suggesting that there is a correlation between intestinal microorganisms and uric acid and urinary nitrogen levels.

[0083] The study, “Shuting Tong et al, The role of gut microbiota in gout: Is gut microbiota a potential target for gout treatment [J]. Front Cell Infect Microbiol. 2022 Nov 24;12:1051682,” demonstrates that the gut microbiota is crucial for host health, regulating metabolism, endocrine function, and the immune system. For example, the gut microbiota plays a role in the occurrence and development of gout. During the pathogenesis of gout, changes in the composition and metabolism of the gut microbiota lead to abnormal uric acid degradation, increased uric acid production, release of pro-inflammatory mediators, and damage to the intestinal barrier. Therefore, targeting the gut microbiota for uric acid level regulation has medical value.

[0084] Clostridium difficile and Salmonella are common intestinal pathogens, while Bacillus subtilis is a common beneficial intestinal probiotic. Clostridium difficile infection is often accompanied by dysbiosis, especially a reduction in short-chain fatty acid-producing bacteria, which are closely related to uric acid metabolism. Salmonella infection can trigger acute intestinal inflammation, damage the intestinal epithelial barrier, promote endotoxin entry into the bloodstream, induce systemic inflammation, and may exacerbate hyperuricemia. Bacillus subtilis, as a probiotic, may improve hyperuricemia by strengthening the intestinal barrier, regulating immunity, and reducing inflammatory factors such as TNF-α.

[0085] Based on the above reasons, this invention explored the effects of various agents on intestinal pathogens such as Clostridium difficile, Salmonella, and Bacillus subtilis. The results showed that the agents in the example groups selectively inhibited Clostridium difficile and Salmonella (inhibition rate ≥73.33%), but had no significant inhibitory effect on the probiotic Bacillus subtilis (inhibition rate ≤2.33%). Therefore, the agents of this invention can reduce inflammation, improve immunity, and reduce the occurrence of hyperuricemia from the perspective of regulating intestinal flora. Control group 1 was simply Dioscorea nipponica powder, and its antibacterial effect was significantly lower than that of the example groups. Control group 2 was a positive control of total saponins from Dioscorea nipponica; although it is an extract and has a significant effect on reducing uric acid and urea nitrogen, its antibacterial effect was not as good as that of the example groups. Control group 3 was a low dose of sodium chloride, and control group 4 was a low dose of zinc sulfate, neither of which had a significant antibacterial effect. Control group 5 was a combination of Dioscorea nipponica and zinc sulfate, and its antibacterial effect was slightly worse than that of the example groups.

[0086] It should be noted that when numerical ranges are involved in this invention, it should be understood that both endpoints of each numerical range and any value between the two endpoints can be selected. Since the steps and methods used are the same as in the embodiments, preferred embodiments are described in this invention to avoid redundancy. Although preferred embodiments of this invention have been described, those skilled in the art, once they understand the inventive concept of this invention, can make other changes and modifications to these embodiments, and all such changes and modifications fall within the scope of this invention.

[0087] Obviously, those skilled in the art can make various modifications and variations to this invention without departing from its spirit and scope. If such modifications and variations fall within the scope of equivalents of this invention, then this invention also intends to include these modifications and variations.

Claims

1. A composition containing *Dioscorea nipponica* for treating hyperuricemia, characterized in that, The composition containing *Dioscorea nipponica* is prepared from *Dioscorea nipponica*, sodium chloride, and zinc sulfate, with a mass ratio of *Dioscorea nipponica*, sodium chloride, and zinc sulfate of 1000:1~2:0~1.

2. The composition containing *Dioscorea nipponica* for treating hyperuricemia according to claim 1, characterized in that, The mass ratio of the smilax, sodium chloride, and zinc sulfate is 1000:1.5:0.

5.

3. The composition containing *Dioscorea nipponica* for treating hyperuricemia according to claim 1, characterized in that, Prepared according to the following method: Dry the pangolin and grind it into powder; Prepare the powdered *Dioscorea nipponica*, sodium chloride, and zinc sulfate according to the mass ratio; Mix thoroughly to obtain a composition containing *Dioscorea nipponica* for the treatment of hyperuricemia.

4. The composition containing *Dioscorea nipponica* for treating hyperuricemia according to claim 3, characterized in that, The powder of *Dioscorea nipponica* is 100-200 mesh.

5. The application of the composition containing *Dioscorea nipponica* according to claim 1 in the preparation of a drug for treating hyperuricemia, characterized in that, A drug for treating hyperuricemia is prepared by using the composition containing *Dioscorea nipponica* for treating hyperuricemia as the sole active ingredient, combined with pharmaceutically acceptable excipients.

6. The application of the composition containing *Dioscorea nipponica* according to claim 5 in the preparation of a drug for treating hyperuricemia, characterized in that, Pharmaceutically acceptable excipients include lubricants, adhesives, or surfactants.

7. The application of the composition containing *Dioscorea nipponica* according to claim 6 in the preparation of a drug for treating hyperuricemia, characterized in that... The lubricant is stearic acid; The adhesive is polyvinyl ketone; The surfactant is sodium dodecyl sulfate or polyoxyethylene fatty acid ester.

8. The application of the composition containing *Dioscorea nipponica* according to claim 5 in the preparation of a drug for treating hyperuricemia, characterized in that, The drugs mentioned above for treating hyperuricemia refer to drugs that lower uric acid and urinary nitrogen levels.

9. The application of the composition containing *Dioscorea nipponica* according to claim 8 in the preparation of a drug for treating hyperuricemia, characterized in that, The uric acid content refers to its content in serum; The urinary nitrogen content refers to its concentration in serum.

10. The use of the composition containing *Dioscorea nipponica* according to claim 5 in the preparation of a drug for treating hyperuricemia, characterized in that, The drugs mentioned above for treating hyperuricemia are those that inhibit the growth of intestinal pathogens.

Images