Application of quercetagetin in preparation of medicine for preventing hyperuricemia and / or hyperuricemia-related diseases

Abstract

The invention relates to the field of biological medicine, in particular to application of quercetagetin in preparation of medicine for preventing hyperuricemia and / or hyperuricemia related diseases. The invention discovers that quercetagetin has a prevention or improvement effect on hyperuricemia and related diseases for the first time. The result of the specific embodiment of the invention shows that the quercetagetin has a good uric acid reducing effect, obviously reduces the activity of uric acid generation participating enzyme, regulates the expression of uric acid transporter mRNA, improves liver injury and kidney injury caused by hyperuricemia, can prevent or relieve hyperuricemia or hyperuricemia-related diseases, and has a good application prospect. The composition can be used for preparing oral tablets and effervescent tablets.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to the use of quercetin in the preparation of medicines for preventing hyperuricemia and / or hyperuricemia-related diseases. Background Art

[0002] Uric acid (UA) in healthy people is in dynamic balance, with 600-700 mg produced or excreted every day. However, due to the influence of subjective or objective factors, the dynamic balance of uric acid in the body will be broken, resulting in excessive uric acid production or reduced uric acid excretion, which manifests as elevated serum uric acid levels. When the fasting serum uric acid level of men is higher than 420 μmol / L and the fasting serum uric acid level of women is higher than 360 μmol / L, they will be diagnosed with hyperuricemia (HUA). Hyperuricemia can be divided into primary hyperuricemia and secondary hyperuricemia. Primary hyperuricemia has a certain degree of heredity; while secondary hyperuricemia is mostly caused by purine metabolism disorders caused by multiple factors such as excessive intake of purine, fructose or beer in the later stage and kidney disease, thereby inducing hyperuricemia. Currently, most patients with hyperuricemia belong to the latter.

[0003] Hyperuricemia patients mainly show excessive uric acid production or reduced uric acid excretion or a combination of the two, and more than 90% of patients show insufficient renal excretion. The excretion of uric acid in the kidney depends on the transport of uric acid transporters. Among them, uric acid transporter 1 (URAT1), a member of the organic anion transporter (OAT) family, is a typical uric acid reabsorption protein. It is highly expressed on the luminal membrane side of the proximal tubular epithelial cells of the renal cortex and can play a 50% reabsorption role in the proximal tubules of the renal cortex. The voltage-driven uric acid transporter glucose transporter 9 (GLUT9) is located on the basolateral membrane of the proximal tubule and plays an equally important reabsorption role as URAT1 in the process of uric acid reabsorption. The low-affinity, high-capacity adenosine triphosphate binding transporter G2 (ABCG2) has the function of excreting uric acid, and its activity can determine the intracellular uric acid level. In addition to innate purine metabolism disorders or acquired factors, studies have shown that environmental factors are also one of the causes of hyperuricemia.

[0004] At present, the treatment of hyperuricemia mainly involves taking drugs to inhibit the activity of enzymes related to purine metabolism to inhibit uric acid production or promote uric acid excretion to reduce the uric acid level in the body. Drugs that inhibit purine metabolism-related enzymes include allopurinol, febuxostat, and topiroxetine; drugs that promote uric acid excretion include benzbromarone, lesinade, etc. Among them, allopurinol and benzbromarone are the most commonly used drugs to treat hyperuricemia. Allopurinol achieves the effect of reducing uric acid levels by inhibiting the conversion of hypoxanthine to xanthine and the conversion of xanthine to uric acid; benzbromarone promotes uric acid excretion mainly by inhibiting the reabsorption of uric acid by the renal tubules, and is a drug to fight gout. Patients with asymptomatic hyperuricemia should first choose benzbromarone as a treatment drug, or regulate the uric acid level in the body through diet and exercise.

[0005] Marigold is widely cultivated and is used in large quantities to extract lutein, but the utilization rate of marigold residue is low. Quercetin (QG) can be extracted from marigold residue. Quercetin, also known as hexahydroxyflavone, is the main plant metabolite in marigold. As a polyhydroxyphenol molecule with multiple hydrogen donor substituents, quercetin has rich biological activities, including strong antioxidant and free radical scavenging abilities, as well as immunomodulatory, anti-inflammatory and antibacterial effects, and exhibits antioxidant and anti-inflammatory advantages that are stronger than quercetin. There are currently no reports that quercetin can prevent and improve hyperuricemia and its related diseases. Summary of the invention

[0006] The purpose of the present invention is to provide the use of quercetin in the preparation of a drug for preventing hyperuricemia and / or hyperuricemia-related diseases, so as to solve the problems existing in the above-mentioned prior art. The present invention first discovered that quercetin has a preventive or ameliorative effect on hyperuricemia and its related diseases.

[0007] To achieve the above object, the present invention provides the following solutions:

[0008] The present invention provides the use of quercetin in the preparation of a medicine for preventing hyperuricemia and / or hyperuricemia-related diseases.

[0009] Preferably, the drug reduces the activity of enzymes involved in uric acid production and regulates the expression of uric acid transporter mRNA, thereby preventing hyperuricemia and / or hyperuricemia-related diseases;

[0010] And / or, the hyperuricemia-related disease includes ventilation.

[0011] Further preferably, the uric acid transporters include ABCG2, GLUT9 and URAT1.

[0012] Further preferably, the enzymes include adenosine deaminase, purine nucleotide phosphorylase, xanthine oxidase and phosphoribosyl pyrophosphate synthetase.

[0013] The present invention provides a medicine for preventing hyperuricemia and / or hyperuricemia-related diseases. The medicine comprises quercetin and auxiliary materials.

[0014] Preferably, the auxiliary materials include three of sodium carboxymethyl starch, magnesium stearate, microcrystalline cellulose, acid-base mixture and dextrin;

[0015] and / or, the hyperuricemia-related diseases include ventilation;

[0016] And / or, the acid source in the acid-base mixture includes malic acid, tartaric acid, citric acid, citric acid and fumaric acid, and the alkali source includes sodium bicarbonate; the mass ratio of the acid source to the alkali source in the acid-base mixture is 0.8:1.

[0017] Further preferably, the dextrin comprises maltodextrin.

[0018] Further preferably, the acid source in the acid-base mixture is citric acid, and the alkali source is sodium bicarbonate.

[0019] In a specific embodiment of the present invention, the sodium carboxymethyl starch is a disintegrant; the magnesium stearate is a lubricant; the microcrystalline cellulose is a filler; the acid-base mixture is a disintegrant; and the dextrin is a filler.

[0020] When preparing the quercetin oral tablets (ordinary oral tablets) of the present invention, the sodium carboxymethyl starch is a disintegrant; the magnesium stearate is a lubricant; and the microcrystalline cellulose is a filler. Sodium carboxymethyl starch has very good absorption and swelling properties, good fluidity and plasticity, and is suitable for the preparation of various poorly soluble drugs, helping tablets to effectively increase the disintegration rate and promote the dissolution of drugs in tablets. Magnesium stearate has good lubricity and strong anti-stickiness, can reduce the friction between particles, and improve the compressibility and fluidity of the powder.

[0021] In the preparation of the quercetin effervescent tablets of the present invention, the acid-base mixture is a disintegrant; the magnesium stearate is a lubricant; and the dextrin is a filler. The present invention uses an acid source (citric acid) and an alkali source (sodium bicarbonate) as disintegrants to give full play to the characteristic of the effervescent tablet that it disintegrates quickly, which is superior to ordinary oral tablets.

[0022] The mass percentage of the effective ingredient (quercetin) in the quercetin oral tablet and the quercetin effervescent tablet provided by the present invention is 60% and 20% respectively. Moreover, the quercetin oral tablet and the quercetin effervescent tablet provided by the present invention have uniform texture, suitable hardness, good safety, and good application prospects.

[0023] Preferably, when the excipients include the sodium carboxymethyl starch, the magnesium stearate and the microcrystalline cellulose, the mass percentage of quercetin in the drug is 60%, the mass percentage of sodium carboxymethyl starch is 8%, the mass percentage of magnesium stearate is 0.4%, and the mass percentage of microcrystalline cellulose is 31.6%;

[0024] When the excipients include the acid-base mixture, the magnesium stearate and the dextrin, the mass percentage of quercetin in the drug is 20%, the mass percentage of the acid-base mixture is 40%, the mass percentage of magnesium stearate is 1%, and the mass percentage of dextrin is 39%.

[0025] Further preferably, the drug reduces the activity of enzymes involved in uric acid production and regulates the expression of uric acid transporter mRNA, thereby preventing hyperuricemia and / or hyperuricemia-related diseases;

[0026] And / or, the hyperuricemia-related disease includes ventilation.

[0027] Further preferably, the uric acid transporters include ABCG2, GLUT9 and URAT1.

[0028] Further preferably, the enzymes include adenosine deaminase, purine nucleotide phosphorylase, xanthine oxidase and phosphoribosyl pyrophosphate synthetase.

[0029] The present invention provides the use of quercetin in preparing a medicine for treating hyperuricemia and / or hyperuricemia-related diseases.

[0030] Preferably, the drug reduces the activity of enzymes involved in uric acid production, regulates the expression of uric acid transporter mRNA, improves liver damage and kidney damage, and achieves the effect of treating hyperuricemia and / or hyperuricemia-related diseases;

[0031] And / or, the hyperuricemia-related disease includes ventilation.

[0032] Further preferably, the uric acid transporters include ABCG2, GLUT9 and URAT1.

[0033] Further preferably, the enzymes include adenosine deaminase, purine nucleotide phosphorylase, xanthine oxidase and phosphoribosyl pyrophosphate synthetase.

[0034] The present invention provides a medicine for treating hyperuricemia and / or hyperuricemia-related diseases, and the medicine comprises quercetin and auxiliary materials.

[0035] Preferably, the auxiliary materials include three of sodium carboxymethyl starch, magnesium stearate, microcrystalline cellulose, acid-base mixture and dextrin;

[0036] and / or, the hyperuricemia-related diseases include ventilation;

[0037] And / or, the acid source in the acid-base mixture includes malic acid, tartaric acid, citric acid, citric acid and fumaric acid, and the alkali source includes sodium bicarbonate; the mass ratio of the acid source to the alkali source in the acid-base mixture is 0.8:1.

[0038] Further preferably, the dextrin comprises maltodextrin.

[0039] Further preferably, the acid source in the acid-base mixture is citric acid, and the alkali source is sodium bicarbonate.

[0040] Preferably, when the excipients include the sodium carboxymethyl starch, the magnesium stearate and the microcrystalline cellulose, the mass percentage of quercetin in the drug is 60%, the mass percentage of sodium carboxymethyl starch is 8%, the mass percentage of magnesium stearate is 0.4%, and the mass percentage of microcrystalline cellulose is 31.6%;

[0041] When the excipients include the acid-base mixture, the magnesium stearate and the dextrin, the mass percentage of quercetin in the drug is 20%, the mass percentage of the acid-base mixture is 40%, the mass percentage of magnesium stearate is 1%, and the mass percentage of dextrin is 39%.

[0042] Further preferably, the drug reduces the activity of enzymes involved in uric acid production, regulates the expression of uric acid transporter mRNA, improves liver damage and kidney damage, and achieves the effect of treating hyperuricemia and / or hyperuricemia-related diseases;

[0043] And / or, the hyperuricemia-related disease includes ventilation.

[0044] Further preferably, the uric acid transporters include ABCG2, GLUT9 and URAT1.

[0045] Further preferably, the enzymes include adenosine deaminase, purine nucleotide phosphorylase, xanthine oxidase and phosphoribosyl pyrophosphate synthetase.

[0046] The present invention discloses the following technical effects:

[0047] The present invention has discovered for the first time that quercetin has a preventive or ameliorative effect on hyperuricemia and its related diseases. The results of the specific embodiments of the present invention show that quercetin has a good uric acid-lowering effect, significantly reduces the activity of enzymes involved in uric acid production, regulates the expression of uric acid transporter mRNA, improves liver damage and kidney damage caused by high uric acid, can prevent or alleviate hyperuricemia or hyperuricemia-related diseases, and can be used to prepare quercetin oral tablets and quercetin effervescent tablets. At the same time, since quercetin can be extracted from marigold residues, the present invention is also of great significance to the utilization of marigold residue resources. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0049] Figure 1Figure 2 shows the serum indexes and enzymes involved in liver uric acid production of mice in different treatment groups; a is the BUN content in serum; b is the CRE content in serum; c is the UA content in serum; d is the activity of liver ADA; e is the activity of liver PNP; f is the activity of liver PRPS; g is the activity of liver XOD; NC is C57BL / 6 mice treated with 0.5% sodium carboxymethylcellulose by oral gavage (control group); MC is treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and 0.5% sodium carboxymethylcellulose by oral gavage =C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and allopurinol (20 mg / kg) by gavage (positive control group); Que-100 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (100 mg / kg) by gavage; Que-200 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (100 mg / kg) by gavage. / kg) and quercetin (200 mg / kg) were gavaged into C57BL / 6 mice; Que-400 was gavaged into C57BL / 6 mice with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (400 mg / kg); QG-50 was gavaged into C57BL / 6 mice with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (50 mg / kg); QG-100 was gavaged into C57BL / 6 mice with adenine (50 mg / kg) g), potassium oxonate (200 mg / kg) and quercetin (100 mg / kg) by gavage; QG-200 is C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (200 mg / kg) by gavage; QG-400 is C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (400 mg / kg) by gavage;

[0050] Figure 2Figure 2 is the H&E staining of the liver of mice in different treatment groups; green arrows: watery degeneration of hepatocytes; red arrows: vascular congestion; scale bar is 100 μm; NC is C57BL / 6 mice treated with 0.5% sodium carboxymethylcellulose by gavage only (control group); MC is C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and 0.5% sodium carboxymethylcellulose by gavage (model group); PC is C57BL / 6 mice treated with adenine (50 mg / kg), oxonate potassium (200 mg / kg) and 0.5% sodium carboxymethylcellulose by gavage =C57BL / 6 mice treated with potassium oxazolidinone (200 mg / kg) and allopurinol (20 mg / kg) by gavage (positive control group); Que-100 C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxazolidinone (200 mg / kg) and quercetin (100 mg / kg) by gavage; Que-200 C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxazolidinone (200 mg / kg) and quercetin (200 mg / kg) by gavage 57BL / 6 mice; Que-400 is C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (400 mg / kg) by gavage; QG-50 is C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (50 mg / kg) by gavage; QG-100 is C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (50 mg / kg) by gavage. g / kg) and quercetin (100 mg / kg) were intragastrically treated in C57BL / 6 mice; QG-200 was intragastrically treated in C57BL / 6 mice with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (200 mg / kg); QG-400 was intragastrically treated in C57BL / 6 mice with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (400 mg / kg);

[0051] Figure 3Figure 2 is a graph of liver oxidative stress indicators of mice in different treatment groups; a is liver MDA content; b is liver GSH-Px activity; c is liver SOD activity; d is liver NO content; NC is C57BL / 6 mice treated with 0.5% sodium carboxymethylcellulose by intragastric administration only; NC is C57BL / 6 mice treated with 0.5% sodium carboxymethylcellulose by intragastric administration only (control group); MC is C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and 0.5% sodium carboxymethylcellulose by intragastric administration 57BL / 6 mice (model group); PC was C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and allopurinol (20 mg / kg) by gavage (positive control group); Que-100 was C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (100 mg / kg) by gavage; Que-200 was C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (100 mg / kg) by gavage. g) and quercetin (200 mg / kg) by gavage in C57BL / 6 mice; Que-400 was treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (400 mg / kg) by gavage in C57BL / 6 mice; QG-50 was treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (50 mg / kg) by gavage in C57BL / 6 mice; QG-100 was treated with adenine (50 mg / kg ), potassium oxonate (200 mg / kg) and quercetin (100 mg / kg) by gavage in C57BL / 6 mice; QG-200 is a C57BL / 6 mouse treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (200 mg / kg) by gavage in C57BL / 6 mice; QG-400 is a C57BL / 6 mouse treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (400 mg / kg) by gavage in C57BL / 6 mice;

[0052] Figure 4Figure 2 shows the kidney morphology of mice in different treatment groups; NC is C57BL / 6 mice treated with 0.5% sodium carboxymethylcellulose by gavage (control group); MC is C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and 0.5% sodium carboxymethylcellulose by gavage (model group); PC is C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and allopurinol (20 mg / kg) C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (100 mg / kg) by gavage were used for Que-100; C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (200 mg / kg) by gavage were used for Que-200; C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (200 mg / kg) by gavage were used for Que-400; C57BL / 6 mice were treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (400 mg / kg) by gavage; QG-50 was treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (50 mg / kg) by gavage; QG-100 was treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (50 mg / kg) by gavage. C57BL / 6 mice treated with marigoldin (100 mg / kg) by gavage; QG-200 is C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (200 mg / kg) by gavage; QG-400 is C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (400 mg / kg) by gavage;

[0053] Figure 5H&E staining of the kidneys of mice in different treatment groups; green arrows: hydropic degeneration of renal tubular epithelial cells; blue arrows: tubular dilatation; gray arrows: tubular atrophy; orange arrows: connective tissue hyperplasia; purple arrows: lymphocyte infiltration; scale bar is 100 μm; NC is C57BL / 6 mice treated with 0.5% sodium carboxymethylcellulose by oral gavage only (control group); MC is C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and 0.5% sodium carboxymethylcellulose by oral gavage L / 6 mice (model group); PC is C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and allopurinol (20 mg / kg) by gavage (positive control group); Que-100 is C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (100 mg / kg) by gavage; Que-200 is C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (100 mg / kg) by gavage and quercetin (200 mg / kg) by gavage in C57BL / 6 mice; Que-400 is C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (400 mg / kg) by gavage; QG-50 is C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (50 mg / kg) by gavage; QG-100 is C57BL / 6 mice treated with adenine (50 mg / kg) , potassium oxonate (200 mg / kg) and quercetin (100 mg / kg) were administered to C57BL / 6 mice; QG-200 was administered to C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (200 mg / kg) by oral gavage; QG-400 was administered to C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (400 mg / kg) by oral gavage;

[0054] Figure 6Figure 2 is a graph of renal inflammation indicators in mice under different treatment groups; a is the renal TNF-α content; b is the renal IL-1β content; c is the renal IL-6 content; d is the renal IL-10 content; NC is C57BL / 6 mice treated with only 0.5% sodium carboxymethylcellulose by gavage (control group); MC is C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and 0.5% sodium carboxymethylcellulose by gavage (model group); PC is C57BL / 6 mice treated with adenine ( =100 mg / kg), oxonate potassium (200 mg / kg), and allopurinol (20 mg / kg) were administered to C57BL / 6 mice (positive control group); Que-100 was administered to C57BL / 6 mice with adenine (50 mg / kg), oxonate potassium (200 mg / kg), and quercetin (100 mg / kg) by oral gavage; Que-200 was administered to C57BL / 6 mice with adenine (50 mg / kg), oxonate potassium (200 mg / kg), and quercetin (200 mg / kg) by oral gavage. ) were administered to C57BL / 6 mice by gavage; Que-400 was administered to C57BL / 6 mice by gavage with adenine (50 mg / kg), potassium oxonate (200 mg / kg), and quercetin (400 mg / kg); QG-50 was administered to C57BL / 6 mice by gavage with adenine (50 mg / kg), potassium oxonate (200 mg / kg), and quercetin (50 mg / kg); QG-100 was administered to C57BL / 6 mice by gavage with adenine (50 mg / kg), potassium oxonate (2 00mg / kg) and quercetin (100mg / kg) were intragastrically treated in C57BL / 6 mice; QG-200 was intragastrically treated in C57BL / 6 mice with adenine (50mg / kg), potassium oxonate (200mg / kg) and quercetin (200mg / kg); QG-400 was intragastrically treated in C57BL / 6 mice with adenine (50mg / kg), potassium oxonate (200mg / kg) and quercetin (400mg / kg);

[0055] Figure 7Figure 2 is the relative expression of uric acid transporter in the kidney of mice in different treatment groups; a is the renal URAT1 mRNA level; b is the renal GLUT9 mRNA level; c is the renal ABCG2 mRNA level; NC is C57BL / 6 mice treated with 0.5% sodium carboxymethylcellulose by gavage only (control group); MC is C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and 0.5% sodium carboxymethylcellulose by gavage (model group); PC is C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and allopurinol (20 mg / kg) by gavage 7BL / 6 mice (positive control group); Que-100 is C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (100 mg / kg) by gavage; Que-200 is C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (200 mg / kg) by gavage; Que-400 is C57BL / 6 mice treated with adenine ( =QG-50 is a C57BL / 6 mouse treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (400 mg / kg) by gavage; QG-100 is a C57BL / 6 mouse treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin by gavage QG-200 was administered to C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (200 mg / kg) by gavage; QG-400 was administered to C57BL / 6 mice treated with adenine (50 mg / kg), potassium oxonate (200 mg / kg) and quercetin (400 mg / kg) by gavage. DETAILED DESCRIPTION

[0056] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0057] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0058] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0059] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0060] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0061] Unless otherwise specified, the methods used in the present invention are all methods well known to those skilled in the art, that is, the contents not described in detail in the embodiments are all carried out according to the prior art in the art.

[0062] Quercetin is referred to as QG; quercetin is provided by Chenguang Biotechnology Group Co., Ltd. with a purity of 95.46%.

[0063] Example 1 Study on the intervention of quercetin on hyperuricemia in mice

[0064] 1. Materials and methods

[0065] 1.1 Experimental Animals

[0066] SPF male C57BL / 6 mice, 6 weeks old, 90 (license number: SCXK (Beijing) 2024-0001), were purchased from Sibeifu (Beijing) Biotechnology Co., Ltd. All mice were kept in a well-ventilated environment with a temperature of 24-26°C, a humidity of 40%-60%, and a light-dark cycle of 12h (light time 12h: dark time 12h). During this period, all mice had independent access to food and water. This experiment was approved by the Experimental Animal Ethics Committee of Hebei Agricultural University (ethics approval code: 2023197; approval date: 2023.12.15)

[0067] 1.2. Establishment and intervention of hyperuricemia mouse model

[0068] 90 C57BL / 6 mice were randomly divided into 10 groups after adapting to a well-ventilated environment with a temperature of 24-26℃, a humidity of 40%-60%, and a light-dark cycle of 12h for one week. Except for the mice in the NC group, the mice in the other groups were treated with adenine (50mg / kg) and potassium oxonate (200mg / kg) by gavage at 9:00 every day. The mice in the NC group were treated with the same dose of 0.5% sodium carboxymethylcellulose by gavage for 21 consecutive days. Starting from the 8th day, the NC and MC groups were treated with the same dose of 0.5% sodium carboxymethylcellulose by gavage at 14:00 every day; the PC group was treated with allopurinol (20mg / kg) by gavage. The Que group was treated with three doses of quercetin (100, 200, 400 mg / kg) by gavage, which were recorded as Que-100, Que-200, and Que-400, respectively. The QG group was treated with four doses of quercetin (50, 100, 200, 400 mg / kg) by gavage, which were recorded as QG-50, QG-100, QG-200, and QG-400, respectively. This was carried out for 14 consecutive days, and the experiment lasted for a total of 21 days.

[0069] Before the end of the experiment, the animals were fasted for 12 hours but not water, and the eyeballs were removed to collect blood, which was then allowed to stand at room temperature for 1 hour. The blood samples were then centrifuged at 3000 rpm for 10 minutes to obtain serum. After the cervical vertebrae of the mice were dislocated, the mice were dissected, and the kidneys, liver, spleen, thymus, pancreas, duodenum, cecum and other tissues were taken. The liver and one of the kidneys were immediately placed in tissue fixative for subsequent pathological histological examination, and the other tissue samples were quickly frozen in liquid nitrogen. Finally, the serum and tissue samples were stored at -80°C for subsequent analysis.

[0070] 1.3 Analysis of Physiological Indexes of Mouse Serum and Tissues

[0071] Commercial kits were used to detect the levels of uric acid (UA), creatinine (CRE), and urea nitrogen (BUN) in serum, the activities of adenosine deaminase (ADA), purine nucleotide phosphorylase (PNP), phosphoribosyl pyrophosphate synthetase (PRPS), xanthine oxidase (XOD), glutathione peroxidase (GSH-Px), and superoxide dismutase (SOD) in liver, and the levels of malondialdehyde (MDA) and nitric oxide (NO) in kidney, as well as the levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), and interleukin-10 (IL-10) in kidney.

[0072] Take 100 mg of kidney and liver, add 900 mL of normal saline, break into homogenate, centrifuge at 3000 rpm at 4°C for 10 min, collect the supernatant, and measure the corresponding indicators.

[0073] After the mice were dissected, the kidneys and livers were fixed and embedded, and paraffin sections and frozen sections were prepared. After hematoxylin and eosin (H&E) staining, the tissue morphology was observed using an optical microscope.

[0074] 1.4 Analysis of uric acid transporter-related gene expression levels in mouse kidney

[0075] (1) Design and synthesis of primers for uric acid transporter-related genes

[0076] The appropriate primer sequences were searched and verified according to the primer design principles on the NCBI website, and finally the primers were synthesized by General Biotechnology Co., Ltd. The primers are shown in Table 1.

[0077] Table 1 Primer sequences

[0078]

[0079] (2) Extraction of total RNA from mouse kidney tissue

[0080] Mouse kidneys frozen at -80°C were taken, and RNA from the kidney tissues of mice in different treatment groups was extracted strictly according to the instructions of RNAEasy Fast Animal Tissue / Cell Total RNA Extraction Kit (Beijing Tiangen Biochemical Technology Co., Ltd.). The extracted RNA was measured by UV-visible spectrophotometer for OD 260 / OD 280 The ratio was used to determine the concentration and purity of RNA.

[0081] (3) RNA reverse transcription

[0082] According to the operating steps provided by the FastKing One-Step Genomic cDNA First-Strand Synthesis Premix Kit provided by Beijing Tiangen Biochemical Technology Co., Ltd., the extracted RNA was reverse transcribed into cDNA. The reverse transcription reaction system and procedure are shown in Tables 2 and 3.

[0083] Table 2 Reverse transcription system

[0084] Composition Usage 5×FastKing-RTSuperMix 4μL Total RNA 50ng-2μg <![CDATA[RNase-FreeddH2O]]> Make up to 20 μL

[0085] Table 3 Reverse transcription procedures

[0086] Reaction temperature Reaction time illustrate 42℃ 15min Genome removal and reverse transcription reaction 95℃ 3min Enzyme inactivation process

[0087] (4) Real-time fluorescence quantitative PCR

[0088] Prepare the Real-Time reaction solution on ice according to the reaction system in Table 4.

[0089] Table 4 qPCR reaction system

[0090] Composition 20μL system 2×FastRealqPCRPreMix(SYBRGreen) 10μL Forward primer (10 μM) 0.6μL Reverse primer (10 μM) 0.6μL cDNA template 1μL <![CDATA[50×ROXReferenceDye △ ]]> 0.4μL <![CDATA[RNase-FreeddH2O]]> Up to 20 μL

[0091] The Real-Time PCR reaction was performed according to the reaction program in Table 5.

[0092] Table 5 qPCR reaction procedure

[0093]

[0094] Adoption 2 -ΔΔCT Methods The relative expression levels were calculated.

[0095] 1.5. Preparation of quercetin oral tablets (ordinary oral tablets)

[0096] Sodium carboxymethyl starch (disintegrant) has very good absorption and swelling properties, good fluidity and plasticity, and is suitable for the preparation of various poorly soluble drugs, helping tablets to effectively increase the disintegration rate and promote the dissolution of drugs in tablets. Magnesium stearate (lubricant) has good lubricity and strong anti-stickiness, which can reduce the friction between particles and improve the compressibility and fluidity of powders. Microcrystalline cellulose is a filler with high mechanical strength and good fluidity.

[0097] (1) Preparation of quercetin oral tablets

[0098] Weigh the main ingredients and auxiliary materials according to the proportion, mix them evenly, pass them through a 60-mesh sieve, and press them into tablets using a tablet press with a diameter of 8 mm.

[0099] The addition amount of sodium carboxymethyl starch was fixed at 6wt.%, the addition amount of magnesium stearate was 0.6wt.%, and the rest was filled with microcrystalline cellulose. The hardness, disintegration time, and dispersion uniformity were used as the standards to explore the effects of different addition amounts of quercetin (40wt.%, 50wt.%, 60wt.%) on the quality of quercetin oral tablets. The addition amount of sodium carboxymethyl starch was fixed at 50wt.%, the addition amount of magnesium stearate was 0.6wt.%, and the rest was filled with microcrystalline cellulose. The hardness, disintegration time, and dispersion uniformity were used as the standards to explore the effects of different addition amounts of sodium carboxymethyl starch (4wt.%, 6wt.%, 8wt.%) on the quality of quercetin oral tablets. The addition amount of the main ingredient quercetin was fixed at 50wt.%, the addition amount of sodium carboxymethyl starch was 6wt.%, and the rest was filled with microcrystalline cellulose. The hardness, disintegration time and dispersion uniformity were used as standards to explore the effect of different magnesium stearate addition amounts (0.4wt.%, 0.6wt.%, 1.0wt.%) on the quality of quercetin oral tablets.

[0100] (2) Testing of properties of quercetin oral tablets

[0101] Determination of the hardness of oral tablets: Set the parameters as follows: TPA mode; P / 2 test probe; strain 50%, trigger force 2N; speeds before, during and after the test are 6mm / s, 0.5mm / s and 30mm / s respectively.

[0102] Method for checking weight differences of oral tablets: According to the weight difference detection method for tablets in the 2020 edition of the Pharmacopoeia of the People's Republic of China, accurately weigh the total weight of 20 randomly selected oral tablets to obtain the average tablet weight, then accurately weigh each oral tablet, and compare the weight of each tablet with the average tablet weight. According to the provisions in Table 6, no more than 2 tablets shall exceed the weight difference limit, and no tablet shall exceed the limit by 1 times.

[0103] Table 6 Weight Difference Limits

[0104] Average tablet weight or stated tablet weight Weight Difference Limit 0.30g or less ±7.5％ 0.30g and above ±5％

[0105] Determine the disintegration time of oral tablets: According to the disintegration time test method for tablets in the 2020 edition of the "Pharmacopoeia of the People's Republic of China" (General Rule 0921), take 6 oral tablets, place them in the hanging basket of the disintegrator, start the instrument and check, and each tablet should be completely disintegrated within 15 minutes. If one tablet cannot be completely disintegrated, another 6 tablets should be taken for retesting, and all should meet the requirements. Among them, the inner diameter of the stainless steel wire mesh is 2mm, the water temperature is 37°C, the screen is 25mm from the bottom of the beaker when the hanging basket drops to the lowest point, and the screen is 15mm below the water surface when the hanging basket rises to the highest point, and the top of the hanging basket cannot be immersed in water.

[0106] Determination of the dispersion uniformity of oral tablets: According to the disintegration time limit test method for tablets in the 2020 edition of the Pharmacopoeia of the People's Republic of China (General Rule 0921), the inner diameter of the stainless steel wire mesh is replaced with 710μm, the water temperature is 15℃-25℃, and 6 oral tablets should all disintegrate and pass through the sieve within 3 minutes. If a small amount cannot pass through the sieve, but has softened into a light weight and has no hard core, it meets the requirements.

[0107] Moisture content of oral tablets: Determine according to the direct drying method in GB 5009.3-2016 National Food Safety Standard Determination of Moisture in Food.

[0108] Microbiological determination of oral tablets: The total colony count is determined in accordance with GB 4789.2-2022 "National Food Safety Standard Food Microbiology Examination Determination of Total Colony Count"; the coliform group is determined in accordance with GB 4789.3-2016 "National Food Safety Standard Food Microbiology Examination Coliform Group Count".

[0109] 1.6. Preparation of quercetin effervescent tablets

[0110] Effervescent tablets use acid sources and alkali sources as disintegrants to achieve their rapid disintegration advantage over oral tablets. The acid sources mainly include malic acid, tartaric acid, citric acid, citric acid, fumaric acid, etc. Citric acid is selected as the acid source for effervescent tablets, sodium bicarbonate as the alkali source for effervescent tablets, dextrin as the filler for effervescent tablets, and magnesium stearate as the lubricant for effervescent tablets.

[0111] (1) Preparation of quercetin effervescent tablets

[0112] Weigh the main ingredients and auxiliary materials according to the proportion, mix them evenly, pass them through a 60-mesh sieve, and press them into tablets using a tablet press with a diameter of 12 mm.

[0113] After mixing the acid source and the alkali source at a mass ratio of 0.4:1, 0.8:1, and 1.6:1, respectively, the appropriate acid-base ratio was determined by the gas production and acidity to obtain an acid-base mixture. Weigh 0.4g of the acid-base mixture and put it in a test tube filled with 50mL of deionized water at 20°C. When no bubbles are generated in the solution, the disintegration is complete. Weigh the total mass of the test tube and the acid-base mixture before and after dissolution. The reduction is the gas production. Randomly select 10 people to taste the acidity.

[0114] The addition amount of the excipient disintegrant (acid-base mixture) was fixed at 30wt.%, the addition amount of magnesium stearate was 0.6wt.%, and the rest was filled with dextrin. The hardness, disintegration time, and appearance state were used as the standards to explore the effects of different quercetin addition amounts (20wt.%, 40wt.%, 60wt.%) on the quality of quercetin effervescent tablets. The addition amount of quercetin was fixed at 40wt.%, the addition amount of magnesium stearate was 0.6wt.%, and the rest was filled with dextrin. The hardness, disintegration time, and appearance state were used as the standards to explore the effects of different disintegrant addition amounts (20wt.%, 30wt.%, 40wt.%) on the quality of quercetin effervescent tablets. The quercetin addition amount was fixed at 40wt.%, the disintegrant addition amount was 30wt.%, and the rest was filled with dextrin. The hardness, disintegration time and appearance were used as standards to explore the effects of different amounts of mesostearate addition (0.4wt.%, 0.6wt.%, 1wt.%) on the quality of quercetin effervescent tablets.

[0115] (2) Property test of quercetin effervescent tablets

[0116] Determine the hardness of effervescent tablets: set the parameters as follows: TPA mode; P / 2 test probe; strain 50%, trigger force 2N; the speeds before, during and after the test are 6mm / s, 0.5mm / s and 30mm / s respectively.

[0117] Method for checking weight differences of effervescent tablets: According to the weight difference detection method for tablets in the 2020 edition of the Pharmacopoeia of the People's Republic of China, accurately weigh the total weight of 20 randomly selected oral tablets to obtain the average tablet weight, then accurately weigh each oral tablet, and compare the weight of each tablet with the average tablet weight. According to the provisions in Table 7, no more than 2 tablets shall exceed the weight difference limit, and no tablet shall exceed the limit by 1 times.

[0118] Table 7 Weight Difference Limits

[0119] Average tablet weight or stated tablet weight Weight Difference Limit 0.30g or less ±7.5％ 0.30g and above ±5％

[0120] Determine the disintegration time of effervescent tablets: According to the disintegration time test method for tablets in the 2020 edition of the Pharmacopoeia of the People's Republic of China (General Rule 0921), take 6 oral tablets, place them in the basket of the disintegrator, start the instrument and check, and all tablets should be completely disintegrated within 15 minutes. If one tablet cannot be completely disintegrated, another 6 tablets should be retested, and all should meet the requirements. The inner diameter of the stainless steel wire mesh is 2mm, the water temperature is 37°C, the screen is 25mm from the bottom of the beaker when the basket drops to the lowest point, and the screen is 15mm below the water surface when the basket rises to the highest point, and the top of the basket cannot be immersed in water.

[0121] Moisture content of effervescent tablets: Determine by direct drying method in accordance with GB 5009.3-2016 National Food Safety Standard - Determination of Moisture in Food.

[0122] Microbiological determination of effervescent tablets: The total colony count is determined in accordance with GB 4789.2-2022 "National Food Safety Standard Food Microbiology Examination Determination of Total Colony Count"; the coliform group is determined in accordance with GB 4789.3-2016 "National Food Safety Standard Food Microbiology Examination Coliform Count".

[0123] 1.7 Data Analysis

[0124] Experimental data are expressed as mean ± standard deviation (SD) and processed using GraphPad Prism 9.5.0. Significant differences were determined by one-way ANOVA. #P<0.05 indicates that the difference is considered statistically significant compared with the NC group. *P<0.05 indicates that the difference is considered statistically significant compared with the MC group. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 indicate that the difference is statistically significant compared with the MC group; #P<0.05, ##P<0.01, ###P<0.001, ####P<0.0001 indicate that the difference is statistically significant compared with the NC group.

[0125] 2. Test results

[0126] 2.1 Analysis of Physiological Indexes of Mouse Blood and Tissues

[0127] BUN is a metabolic product of protein in the body. It will be reabsorbed in the renal tubules after being filtered through the glomeruli. CRE is an important substance produced by human muscle metabolism and mainly excreted through the glomeruli. To a certain extent, the degree of kidney damage caused by HUA can be measured by the levels of BUN and CRE in serum.

[0128] like Figure 1 As shown in a and b, compared with the NC group, the CRE and BUN levels in the MC group increased significantly by 1.77 times and 2.98 times, and the glomerular filtration capacity decreased; compared with the MC group, the Que-200 group and the Que400 group significantly reduced the CRE and BUN levels in the serum, which were reduced by 32.59%, 52.80%, 31.94%, and 68.79%, respectively; the serum CRE in the four doses of QG groups was significantly reduced by 22.53%, 48.31%, 55.74%, and 59.84%, and the serum BUN was significantly reduced by 32.81%, 45.44%, 58.05%, and 70.69%, which showed that QG can improve renal function and enhance the glomerular filtration capacity. The intervention treatment of the same dose of QG and quercetin, among which the intervention of QG was significantly better than quercetin in reducing the serum CRE and BUN levels.

[0129] like Figure 1 As shown in c, compared with the NC group, the serum UA content of the MC group increased significantly by 84.49%, indicating that the hyperuricemia mouse model was successfully established. Compared with the MC group, the serum UA content of the PC group decreased significantly by 40.06%, the serum UA content of the QG-200 group and the QG-400 group decreased significantly by 41.78% and 52.73%, and the serum UA content of the Que-400 group decreased significantly by 45.56%. QG intervention can reduce the serum uric acid level of mice, and the effect is better than that of allopurinol intervention. At the same dose, quercetin is better than quercetin in reducing the serum uric acid content.

[0130] The activities of adenosine deaminase (ADA), purine nucleotide phosphorylase (PNP), xanthine oxidase (XOD), and phosphoribosyl pyrophosphate synthetase (PRPS) are positively correlated with uric acid levels, among which XOD is the rate-limiting enzyme in the purine nucleoside metabolic pathway and plays a leading role in the pathogenesis of hyperuricemia; ADA catalyzes the conversion of adenosine to inosine, which is deribosylated by PNP and converted to hypoxanthine; ADA is proportional to adenosine concentration, and its activity increases in the case of hypoxia and inflammation, and flavonoids can significantly inhibit the activity of ADA. In this embodiment, the intervention of quercetin significantly inhibited ADA activity. PRPS catalyzes the synthesis of nucleotides, and XOD catalyzes the oxidation of hypoxanthine and xanthine to UA.

[0131] like Figure 1As shown in dg, compared with the NC group, the activities of liver ADA, PNP, PRPS, and XOD in the MC group were significantly increased by 96.30%, 58.51%, 69.91%, and 79.71%, respectively. The intervention of QG reduced the activities of liver ADA, PNP, PRPS, and XOD, among which the activities of liver ADA in the QG-100 group, QG-200 group, and QG-400 group were significantly reduced by 24.91%, 51.49%, and 50.72%, respectively. The activities of liver PNP and PRPS in the QG-200 and QG-400 groups were significantly reduced by 24.91%, 51.49%, and 50.72%, respectively. 22.28%, 30.31%, 21.50%, 33.21%; the XOD activity of the four doses of QG groups was significantly reduced by 29.87%, 33.28%, 44.14%, 44.05%, QG in the QG-200 group and the QG-400 group can significantly reduce the liver ADA, PNP, PRPS, and XOD activities; in the intervention treatment of quercetin, only Que-400 can significantly reduce the liver ADA, PNP, PRPS, and XOD activities, which are 44.05%, 36.72%, 45.09%, and 19.75%, respectively. Therefore, the results show that QG can effectively inhibit the activity of enzymes involved in uric acid production in the liver, among which the inhibitory effect on XOD is the best, and the effect of QG is better than that of quercetin.

[0132] Xanthine oxidase (XOD) is the rate-limiting enzyme in the purine nucleoside metabolic pathway and plays a leading role in the pathogenesis of hyperuricemia. 50 mg / kg of quercetin intervention significantly reduced the activity of liver xanthine oxidase by 29.87%, and 400 mg / kg of quercetin intervention significantly reduced the activity of liver xanthine oxidase by 19.75%. Quercetin showed a better dosage advantage

[0133] like Figure 2 As shown in the data, a large number of hepatocytes were found to have hydropic degeneration and occasional vascular congestion in the MC group; quercetin intervention reduced the hydropic degeneration of hepatocytes and vascular congestion; QG intervention also reversed this change, reducing the hydropic degeneration of hepatocytes and vascular congestion as the dose increased.

[0134] like Figure 3As shown in the figure, compared with the NC group, the antioxidant GSH-Px and SOD activities of the MC group were significantly reduced by 47.00% and 22.48%, the NO content was significantly reduced by 49.35%, and the MDA content was significantly increased by 62.86%. Compared with the MC group, the QG intervention treatment significantly increased the antioxidant GSH-Px activities of the QG-100 group, QG-200 group, and QG-400 group by 49.31%, 72.36%, and 81.67%, respectively. The increase in GSH-Px activity specifically catalyzed the reaction of reduced glutathione with reactive oxygen to generate oxidized glutathione, which protected the liver. The lipid peroxide MDA content was significantly reduced by 12.40%, 32.53%, and 43.39%, respectively; the antioxidant SOD activity of the high-dose QG-400 group was significantly increased. It increased by 28.25%. The NO content in the four dose groups of QG increased significantly by 38.63%, 59.32%, 115.05%, and 118.30%. The high-dose QG-400 group can significantly reduce the MDA content, increase the activity of GSH-Px and SOD, and increase the NO content; at the same time, the high-dose quercetin group can also significantly reduce the MDA content, increase the activity of GSH-Px and SOD, and increase the NO content, which changed to 29.82%, 68.06%, 33.31%, and 118.80%, respectively. Therefore, it shows that high uric acid causes oxidative damage to the liver, and QG improves the oxidative damage to the liver by enhancing the liver's antioxidant capacity. Figure 2 Histopathological results showed that QG had the advantage of improving liver oxidative damage.

[0135] like Figure 4 and Figure 5 As shown in the figure, the kidneys of mice in the NC group had a smooth surface, dark red color, and luster, the number of cells and matrix in the glomeruli were uniform, the renal tubular epithelial cells were round and plump, the brush borders were arranged neatly and regularly, there was no obvious proliferation of the renal interstitium between the urinary tubules, and no obvious inflammatory cell infiltration was observed; the kidneys of mice in the MC group were wrinkled, light red in color, and dull, and the renal tubular epithelial cells showed more watery degeneration, more tubular dilatation, more tubular atrophy, a small amount of connective tissue hyperplasia, accompanied by a small amount of lymphocyte infiltration; the intervention of quercetin and QG restored the appearance of the kidneys to that of the NC group, and reduced the watery degeneration of renal tubular epithelial cells, tubular dilatation, tubular atrophy, connective tissue hyperplasia and lymphocyte infiltration.

[0136] like Figure 6As shown, compared with the NC group, the levels of TNF-α, IL-1β, and IL-6 in the MC group were significantly increased by 60.08%, 35.93%, and 45.35%, and the content of anti-inflammatory factor IL-10 was significantly decreased by 33.40%. Compared with the MC group, the levels of TNF-α and IL-1β in the QG-200 group and QG-400 group in the QG intervention treatment were significantly reduced by 34.69%, 42.86%, 17.78%, and 25.16%, respectively; the levels of IL-6 in the four QG dose groups were significantly reduced by 20.32%, 24.04%, 26.60%, and 33.40%, respectively. 40.55%; the anti-inflammatory IL-10 content of QG-100 group, QG-200 group, and QG-400 group increased significantly by 24.06%, 38.38%, and 70.32%, respectively. The QG-200 group and QG-400 group can significantly reduce the content of TNF-α, IL-1β, and IL-6, and increase the content of IL-10; among the three doses of quercetin groups, only the Que-400 group can significantly reduce the content of TNF-α, IL-1β, and IL-6, and increase the content of IL-10, which changed to 32.26%, 19.46%, 38.77%, and 53.47%, respectively. Therefore, it is shown that QG improves kidney inflammation damage caused by high uric acid by reducing the content of pro-inflammatory factors and enhancing anti-inflammatory ability. Combined with Figure 4 and Figure 5 Histopathological results showed that QG had a dose advantage in alleviating renal inflammatory damage.

[0137] 2.2 Analysis of uric acid transporter mRNA levels in the kidneys of mice in each group

[0138] Uric acid transporter 1 (URAT1) is a typical uric acid reabsorption protein, which is highly expressed on the luminal membrane side of the proximal tubule epithelial cells in the renal cortex and can play a 50% reabsorption role in the proximal tubule. The voltage-driven uric acid transporter glucose transporter 9 (GLUT9) is located on the basolateral membrane of the proximal tubule and plays an equally important reabsorption role as URAT1 in the process of uric acid reabsorption. The low-affinity, high-capacity adenosine triphosphate-binding transporter G2 (ABCG2) has the function of excreting uric acid, and its activity can determine the intracellular uric acid level.

[0139] like Figure 7As shown, compared with the NC group, the mRNA expression of secretory protein ABCG2 in the MC group was significantly downregulated by 1.54 times, and the mRNA expressions of reabsorption proteins GLUT9 and URAT1 were significantly upregulated by 4.9 times and 3.44 times; compared with the MC group, the mRNA expression of secretory protein ABCG2 in the Que-400 group was significantly upregulated by 1.41 times; the mRNA expressions of reabsorption proteins GLUT9 and URAT1 in the three doses of quercetin groups were significantly downregulated, by 1.51 times, 2.7 times, 4.28 times, 1.43 times, 1.57 times, and 3.31 times, respectively. The intervention of QG significantly increased the expression of secretory protein ABCG2 mRNA by 1.49-fold and 1.69-fold in the QG-200 and QG-400 groups, and significantly decreased the expression of reabsorption proteins URAT1 and GLUT9 mRNA by 1.38-fold, 2.85-fold, 3.57-fold, 4.32-fold, 1.27-fold, 1.72-fold, 1.96-fold, and 3.45-fold in the four doses of quercetin, respectively, showing a good dose-dependency. Therefore, the results showed that QG promoted uric acid excretion in mice by regulating the expression of uric acid transporter mRNA, and quercetin had a greater dose advantage than quercetin in regulating uric acid transporter mRNA.

[0140] 2.4. Preparation of quercetin oral tablets

[0141] The effects of different addition amounts of quercetin, sodium carboxymethyl starch, and magnesium stearate on the quality of oral tablets are shown in Table 8. It can be found that different addition amounts have the greatest impact on the hardness of oral tablets, and their disintegration time and dispersion uniformity meet the requirements of the 2022 edition of the Pharmacopoeia of the People's Republic of China. According to the hardness, the best addition amount of quercetin is 60wt.%, sodium carboxymethyl starch is 8wt.%, magnesium stearate is 0.4wt.%, and microcrystalline cellulose is 31.6wt.%.

[0142] Table 8 Effects of different substance additions on oral tablets

[0143]

[0144] 2.5. Preparation of quercetin effervescent tablets

[0145] When the acid-base ratio is 0.4:1, 0.8:1, and 1.6:1, the gas production is 0.08±0.01, 0.31±0.02, and 0.56±0.02; when the acid-base ratio is 1.6:1, it is slightly acidic. Finally, based on the results of acidity and gas production, the ratio of citric acid to sodium bicarbonate was determined to be 0.8:1. The effects of different amounts of quercetin, magnesium stearate, maltodextrin, and acid-base disintegrants on the quality of effervescent tablets are shown in Table 9. Different addition amounts have an effect on the disintegration time, hardness, and appearance of the effervescent tablets. According to the hardness, disintegration time, and appearance, the best effervescent tablet ratio was screened out as quercetin 20wt.%, magnesium stearate 1wt.%, disintegrant (citric acid: sodium bicarbonate = 0.8:1) 40wt.%, and maltodextrin 39wt.%.

[0146] Table 9 Effects of different substance addition amounts on effervescent tablets

[0147]

[0148] According to the optimal ratio screened above, quercetin oral tablets and quercetin effervescent tablets were prepared, and the performance survey results are shown in Table 10. The results show that the tablet weight difference, disintegration time, and dispersion uniformity all meet the requirements of the 2020 edition of the Pharmacopoeia of the People's Republic of China. Because quercetin has antibacterial ability, its products also show good antibacterial effects.

[0149] Table 10 Quality Analysis

[0150]

[0151] The conversion coefficient of the dose per kilogram of body weight of mice and humans was 0.11. The mouse dose was converted to the human dose, and the results are shown in Table 11. In animal experiments, when the mouse intervention dose was 200 mg / kg, the serum uric acid level of mice was reduced. Therefore, when the daily dose of adults was 22 mg / kg, the effect of reducing uric acid levels could be achieved. Based on the oral dosage of 60 kg adults, 1.32 g of quercetin should be taken daily, that is, 5 tablets of oral tablets 3 times a day, and about 2 tablets of commercially available effervescent tablets.

[0152] Table 11 Dose conversion

[0153]

[0154] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. Use of quercetin in the preparation of a medicament for preventing hyperuricemia and / or hyperuricemia-related diseases.

2. The use according to claim 1, characterized in that: The drug reduces the activity of enzymes involved in uric acid production and regulates the expression of uric acid transporter mRNA, thereby preventing hyperuricemia and / or hyperuricemia-related diseases. And / or, the hyperuricemia-related disease includes ventilation.

3. A drug for preventing hyperuricemia and / or hyperuricemia-related diseases, characterized in that: The medicine comprises quercetin and auxiliary materials.

4. The drug according to claim 3, characterized in that The auxiliary materials include three of sodium carboxymethyl starch, magnesium stearate, microcrystalline cellulose, acid-base mixture and dextrin; and / or, the hyperuricemia-related diseases include ventilation; And / or, the acid source in the acid-base mixture includes malic acid, tartaric acid, citric acid, citric acid and fumaric acid, and the alkali source includes sodium bicarbonate; the mass ratio of the acid source to the alkali source in the acid-base mixture is 0.8:

1.

5. The drug according to claim 4, characterized in that When the excipients include the sodium carboxymethyl starch, the magnesium stearate and the microcrystalline cellulose, the mass percentage of quercetin in the drug is 60%, the mass percentage of sodium carboxymethyl starch is 8%, the mass percentage of magnesium stearate is 0.4%, and the mass percentage of microcrystalline cellulose is 31.6%; When the excipients include the acid-base mixture, the magnesium stearate and the dextrin, the mass percentage of quercetin in the drug is 20%, the mass percentage of the acid-base mixture is 40%, the mass percentage of magnesium stearate is 1%, and the mass percentage of dextrin is 39%.

6. Use of quercetin in the preparation of drugs for treating hyperuricemia and / or hyperuricemia-related diseases.

7. The use according to claim 6, characterized in that: The drug reduces the activity of enzymes involved in uric acid production, regulates the expression of uric acid transporter mRNA, improves liver damage and kidney damage, and achieves the effect of treating hyperuricemia and / or hyperuricemia-related diseases; And / or, the hyperuricemia-related disease includes ventilation.

8. A drug for treating hyperuricemia and / or hyperuricemia-related diseases, characterized in that: The medicine comprises quercetin and auxiliary materials.

9. The drug according to claim 8, characterized in that The auxiliary materials include three of sodium carboxymethyl starch, magnesium stearate, microcrystalline cellulose, acid-base mixture and dextrin; and / or, the hyperuricemia-related diseases include ventilation; And / or, the acid source in the acid-base mixture includes malic acid, tartaric acid, citric acid, citric acid and fumaric acid, and the alkali source includes sodium bicarbonate; the mass ratio of the acid source to the alkali source in the acid-base mixture is 0.8:

1.

10. The drug according to claim 9, characterized in that When the excipients include the sodium carboxymethyl starch, the magnesium stearate and the microcrystalline cellulose, the mass percentage of quercetin in the drug is 60%, the mass percentage of sodium carboxymethyl starch is 8%, the mass percentage of magnesium stearate is 0.4%, and the mass percentage of microcrystalline cellulose is 31.6%; When the excipients include the acid-base mixture, the magnesium stearate and the dextrin, the mass percentage of quercetin in the drug is 20%, the mass percentage of the acid-base mixture is 40%, the mass percentage of magnesium stearate is 1%, and the mass percentage of dextrin is 39%.

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