A composition for preventing and / or treating hyperuricemia and gout and its application

Abstract

The present invention discloses the use of luteolin, isoquercetin and febuxostat in the preparation of a medicament for preventing and / or treating hyperuricemia and gout. In the composition of luteolin and isoquercetin, the mass ratio of luteolin to isoquercetin is 1:5 to 100. In the solution of the present invention, it is found that luteolin and isoquercetin are used in combination with febuxostat in a specific ratio. On the one hand, it has the effects of inhibiting XOD oxidase and reducing uric acid, and on the other hand, it can antagonize the EP4 receptor to reduce inflammation. Therefore, the dosage of febuxostat can be reduced, the inflammatory factors during the occurrence of gouty arthritis can be inhibited, the acute attack of gouty arthritis can be relieved, and it has a significant effect on the treatment of gouty arthritis without obvious toxic and side effects.

Description

Technical Field

[0001] The present invention belongs to the technical field of medicine, and particularly relates to a composition for preventing and / or treating hyperuricemia and gout and its application. Background Art

[0002] Gout is an arthropathy caused by the deposition of monosodium urate. In recent years, the incidence of hyperuricemia and gout in China has shown an obvious upward trend and a tendency of getting younger, which has greatly increased the burden on patients and society. Persistent hyperuricemia may lead to the onset of gouty arthritis. By reducing the body's urate pool, the onset of gouty arthritis can be reduced. Currently, the anti-gout drugs clinically are mainly small molecule synthetic drugs. Although these drugs have obvious therapeutic effects in reducing serum uric acid levels, they have serious adverse reactions such as allergic reactions and liver and kidney damage. Among them, febuxostat has a good effect on reducing uric acid, but in the first few months of using febuxostat to reduce uric acid, it will instead increase the attack frequency of gouty arthritis, resulting in a decrease in patient compliance and failure to meet the uric acid control standard. Using febuxostat to reduce uric acid during the acute gouty arthritis period may exacerbate the attack degree, prolong the attack time, induce or increase the attack risk, etc. And there are obvious inflammatory factors in gouty arthritis, and febuxostat has no anti-inflammatory effect and poor efficacy. It is usually used in combination with non-steroidal anti-inflammatory drugs, but both non-steroidal anti-inflammatory drugs and febuxostat have gastrointestinal and cardiovascular side effects, and using them together will greatly increase the risks of gastric bleeding and cardiovascular diseases.

[0003] COX2-PGE2-Eps is an important inflammatory pathway. Cyclooxygenases (COXs), also known as prostaglandin endoperoxide reductases, are bifunctional enzymes with both oxidase and catalase activities, and are key enzymes for catalyzing arachidonic acid (AA) into prostaglandin enzymes. When cells are stimulated by inflammation, their expression levels can increase to 80 times that of normal, and then catalyze the synthesis of the inflammatory mediator prostaglandins (PGs) from AA, causing infiltration of inflammatory cells. Prostaglandin E2 (PGE2) is an unsaturated fatty acid with 20 carbons and is widely involved in various pathophysiological processes of the body, including inflammatory reactions and tumor formation. PGE2 binds to the membrane surface PGE2 receptor (EPR) of different cells and mediates a variety of biological effects. There are 4 subtypes of EPR, namely EP1, EP2, EP3, and EP4. Among them, EP4 is a Gs protein-coupled receptor that can activate the cAMP-ERK and PI3K signal transduction pathways. The PGE2 receptor EP4 antagonist can achieve anti-inflammatory and analgesic effects by blocking the downstream signal pathway of PGE2 without affecting the level of prostaglandins, especially PGI2, thereby avoiding the adverse reactions of the kidneys and cardiovascular and cerebrovascular systems caused by COX2 inhibitors.

[0004] Traditional Chinese medicines have high safety. More and more traditional Chinese medicines and their extracts have been found to have a wide range of uric acid-lowering or anti-gout activities and high safety, which has become an important direction for the discovery of new anti-gout drugs.

[0005] Celery seeds contain volatile oils, eucalyptol, phosphomethoxyphenol, umbelliferone, linolenic acid, and flavonoids such as luteolin and apigenin. These active ingredients can inhibit free radicals and have anti-inflammatory effects. Among them, luteolin is commonly used in clinical treatments such as cancer and diabetes. Research shows that due to its anti-inflammatory and antioxidant effects, the NF-κB signaling pathway is one of the classical anti-inflammatory signaling pathways that have been proven.

[0006] Flos Sophorae Immaturus contains chemical components such as rutin, quercetin, triterpenoid saponins (0.4%), betulinol, sophora diol, grapes, glucose, and gluconic aldehyde. Among them, quercetin is a flavonol compound and has good antioxidant and anti-inflammatory effects as an antioxidant free radical. Isoquercetin is a derivative of quercetin and has similar or even higher biological activities and bioavailability to quercetin. Summary of the Invention

[0007] The purpose of the present invention is to provide a composition with good effects in preventing and / or treating hyperuricemia and gout and no toxic side effects.

[0008] Achieving the above purpose includes the following technical solutions.

[0009] The first aspect of the present invention provides an application of luteolin, isoquercetin, and febuxostat in the preparation of a drug for preventing and / or treating hyperuricemia and gout. Among them, in the composition of luteolin and isoquercetin, the mass ratio of luteolin to isoquercetin is 1:5 to 100.

[0010] Above, the luteolin, isoquercetin, and febuxostat can be used in a separate form or a mixed form. The mixed form includes using luteolin and isoquercetin after mixing with febuxostat, or using luteolin and febuxostat after mixing with isoquercetin, or using isoquercetin and febuxostat after mixing with luteolin.

[0011] In some embodiments, in the composition of luteolin and isoquercetin, the mass ratio of luteolin to isoquercetin is 1:8 to 60.

[0012] In some embodiments, in the composition of luteolin and isoquercetin, the mass ratio of luteolin to isoquercetin is 1:8 to 55.

[0013] In some embodiments, in the composition of luteolin and isoquercetin, the mass ratio of luteolin to isoquercetin is 1:9 to 12.

[0014] In some of these embodiments, the gout is gouty arthritis.

[0015] In some of these embodiments, the composition of luteolin and isoquercetin is a composite extract obtained by enzymatic extraction using celery seeds and sophora flower buds as raw materials.

[0016] In some of these embodiments, the added enzyme is 0.5%-10% of the total weight of celery seeds and sophora flower buds.

[0017] In some of these embodiments, the enzymatic extraction method of celery seeds and sophora flower buds includes the following steps:

[0018] Separate the celery seeds and sophora flower buds into powder to obtain celery seed powder and sophora flower bud powder respectively;

[0019] Add the celery seed powder and sophora flower bud powder into a buffer solution, and then add an enzyme for enzymatic extraction to obtain a composite extract.

[0020] In some of these embodiments, the buffer solution is an aqueous solution of citric acid and sodium phosphate dodecahydrate with a pH value of 3.5 to 6.5, and the weight ratio of the total weight of the celery seed powder and sophora flower bud powder to the weight of the buffer solution is 1:10 to 40.

[0021] In some of these embodiments, in the step of obtaining a composite extract by enzymatic extraction, the following steps are included:

[0022] Perform enzymatic extraction at a temperature of 40°C to 60°C for 2h to 4h to obtain an extract;

[0023] Inactivate the enzyme in the extract for 20min to 30min, separate to obtain a filtrate, and dry to obtain a composite extract powder.

[0024] In some of these embodiments, the enzyme is one or more of cellulase, rhamnosidase, naringinase, and pectinase.

[0025] In some of these embodiments, in the raw materials, the celery seeds are 30% to 65%, and the sophora flower buds are 35% - 70%; or in the raw materials, the celery seeds are 81% to 90%, and the sophora flower buds are 10% - 19%.

[0026] The second aspect of the present invention provides a composition for preventing and / or treating hyperuricemia and gout. The active ingredients of the composition are luteolin, isoquercetin, and febuxostat. The active ingredients form the composition in a separate form or a mixed form. Among them, the mass ratio of luteolin to isoquercetin is 1:5 to 100.

[0027] In some of these embodiments, the mass ratio of luteolin to isoquercetin in the composition of luteolin and isoquercetin is 1:8 to 60.

[0028] In some of these embodiments, the mass ratio of luteolin to isoquercetin in the composition of luteolin and isoquercetin is 1:9 to 12.

[0029] In the present invention, it is found that when luteolin and isoquercetin are used in combination with febuxostat in a specific ratio, on the one hand, it has the effects of inhibiting XOD oxidase and reducing uric acid, and on the other hand, it can antagonize the EP4 receptor to reduce inflammation. Thus, the dosage of febuxostat can be reduced, the inflammatory factors during the occurrence of gouty arthritis can be inhibited, the acute attack of gouty arthritis can be relieved, and it has significant curative effects in the treatment of gouty arthritis with no obvious toxic and side effects.

[0030] Furthermore, by enzymatically hydrolyzing celery seeds and sophora flower buds, a composite extract containing luteolin and isoquercetin can be obtained. The extraction rates of luteolin and isoquercetin in this composite extract are higher and more stable, the quality standards are clear, and it has a better effect of reducing uric acid. Compared with the existing ethanol extraction method, the enzymatic hydrolysis extraction method not only avoids the problems of complex components, unstable drug efficacy, unclear safety, and difficult quality control of the prepared extract, but also has the characteristics and advantages of multi-target and multi-effect of traditional prescriptions. Detailed implementation manners

[0031] To facilitate the understanding of the present invention, the present invention will be described more comprehensively below. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, these embodiments are provided to make the understanding of the disclosed content of the present invention more thorough and comprehensive.

[0032] For the experimental methods without specific conditions noted in the following examples, they are generally in accordance with conventional conditions or the conditions recommended by the manufacturers. All kinds of commonly used chemical reagents used in the examples are commercially available products.

[0033] Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as commonly understood by those skilled in the technical field to which the present invention belongs. The terms used in the description of the present invention are only for the purpose of describing specific embodiments and are not used to limit the present invention. The term "and / or" used in the present invention includes any and all combinations of one or more of the related listed items.

[0034] The following are specific embodiments.

[0035] Example 1

[0036] The preparation method of the composite extract containing luteolin and isoquercetin provided in this example includes the following steps:

[0037] The celery seeds and sophora flower buds are respectively pulverized to obtain celery seed powder and sophora flower bud powder with a mesh number of 10.

[0038] Weigh 10 g of celery seed powder and 990 g of sophora flower bud powder, and add a buffer solution that is 10 times the total weight of the celery seed powder and sophora flower bud powder. The buffer solution is an aqueous solution of citric acid and sodium phosphate dodecahydrate, with a pH value of 4; then add 5 ml of cellulase, keep the temperature at 40 °C, enzymatically extract for 2 h, inactivate the enzyme for 20 min, separate to obtain a filtrate, and spray-dry to obtain a composite extract powder of luteolin and isoquercetin.

[0039] Example 2

[0040] The preparation method of the composite extract containing luteolin and isoquercetin provided in this example includes the following steps:

[0041] The celery seeds and sophora flower buds are respectively pulverized to obtain celery seed powder and sophora flower bud powder with a mesh number of 100.

[0042] Weigh 650 g of celery seed powder and 350 g of sophora flower bud powder, and add a buffer solution that is 20 times the total weight of the celery seed powder and sophora flower bud powder. The buffer solution is an aqueous solution of citric acid and sodium phosphate dodecahydrate, with a pH value of 5; then add 10 ml of rhamnosidase, keep the temperature at 60 °C, enzymatically extract for 4 h, inactivate the enzyme for 30 min, separate to obtain a filtrate, and spray-dry to obtain a composite extract powder of luteolin and isoquercetin.

[0043] Example 3

[0044] The preparation method of the composite extract containing luteolin and isoquercetin provided in this example includes the following steps:

[0045] The celery seeds and sophora flower buds are respectively pulverized to obtain celery seed powder and sophora flower bud powder with a mesh number of 200.

[0046] Weigh 350 g of celery seed powder and 650 g of sophora flower bud powder, and add a buffer solution that is 40 times the total weight of the celery seed powder and sophora flower bud powder. The buffer solution is an aqueous solution of citric acid and sodium phosphate dodecahydrate, with a pH value of 6; then add 100 ml of naringinase, keep the temperature at 60 °C, enzymatically extract for 3 h, inactivate the enzyme for 20 min, separate to obtain a filtrate, and spray-dry to obtain a composite extract powder of luteolin and isoquercetin.

[0047] Example 4

[0048] The preparation method of the composite extract containing luteolin and isoquercetin provided in this example includes the following steps:

[0049] The celery seeds and sophora flower buds are respectively pulverized to obtain celery seed powder and sophora flower bud powder with a mesh number of 10.

[0050] Weigh 810 g of celery seed powder and 190 g of sophora flower powder, add a buffer solution that is 10 times the total weight of the celery seed powder and sophora flower powder. The buffer solution is an aqueous solution of citric acid and sodium phosphate dodecahydrate, with a pH value of 4; then add 5 ml of pectinase, keep the temperature at 45 °C, enzymatically extract for 4 h, inactivate the enzyme for 30 min, separate to obtain a filtrate, and spray-dry to obtain a composite extract powder of luteolin and isoquercetin.

[0051] Example 5

[0052] The preparation method of the composite extract containing luteolin and isoquercetin provided in this example includes the following steps:

[0053] Crush celery seeds and sophora flowers separately to obtain celery seed powder and sophora flower powder with a mesh size of 100;

[0054] Weigh 900 g of celery seed powder and 100 g of sophora flower powder, add a buffer solution that is 30 times the total weight of the celery seed powder and sophora flower powder. The buffer solution is an aqueous solution of citric acid and sodium phosphate dodecahydrate, with a pH value of 4; then add 20 ml each of cellulase and rhamnosidase, keep the temperature at 50 °C, enzymatically extract for 4 h, inactivate the enzyme for 30 min, separate to obtain a filtrate, and spray-dry to obtain a composite extract powder of luteolin and isoquercetin.

[0055] Example 6

[0056] The preparation method of the composite extract containing luteolin and isoquercetin provided in this example includes the following steps:

[0057] Crush celery seeds and sophora flowers separately to obtain celery seed powder and sophora flower powder with a mesh size of 200;

[0058] Weigh 990 g of celery seed powder and 10 g of sophora flower powder, add a buffer solution that is 30 times the total weight of the celery seed powder and sophora flower powder. The buffer solution is an aqueous solution of citric acid and sodium phosphate dodecahydrate, with a pH value of 4; then add 30 ml each of naringinase and pectinase, keep the temperature at 55 °C, enzymatically extract for 3 h, inactivate the enzyme for 30 min, separate to obtain a filtrate, and spray-dry to obtain a composite extract powder of luteolin and isoquercetin.

[0059] Comparative Example 7

[0060] The preparation method of the composite extract containing luteolin and isoquercetin provided in this comparative example includes the following steps:

[0061] Crush celery seeds and sophora flowers separately to obtain celery seed powder and sophora flower powder with a mesh size of 10;

[0062] Weigh 810 g of celery seed powder and 190 g of sophora flower powder, add 10 times the total weight of celery seed powder and sophora flower powder of 50% (v / v) ethanol aqueous solution, separate, concentrate and recover ethanol, and spray dry to obtain the composite extract powder of luteolin and isoquercetin.

[0063] Comparative Example 8

[0064] The preparation method of the composite extract containing luteolin and isoquercetin provided in this comparative example includes the following steps:

[0065] Crush celery seeds and sophora flower respectively to obtain celery seed powder and sophora flower powder with a mesh number of 100;

[0066] Weigh 750 g of celery seed powder and 250 g of sophora flower powder, add 10 times the total weight of celery seed powder and sophora flower powder of 60% (v / v) ethanol aqueous solution, extract ultrasonically for 3 times, separate, concentrate and recover ethanol, and spray dry to obtain the composite extract powder of luteolin and isoquercetin.

[0067] Comparative Example 9

[0068] The preparation method of the composite extract containing luteolin and isoquercetin provided in this comparative example includes the following steps:

[0069] Crush celery seeds and sophora flower respectively to obtain celery seed powder and sophora flower powder with a mesh number of 200;

[0070] Weigh 650 g of celery seed powder and 350 g of sophora flower powder, add 10 times the total weight of celery seed powder and sophora flower powder of 70% (v / v) ethanol aqueous solution, extract ultrasonically for 3 times, separate, concentrate and recover ethanol, and spray dry to obtain the composite extract powder of luteolin and isoquercetin.

[0071] Test Example 1 Determination of the contents of luteolin and isoquercetin in the composite extract

[0072] Detection: Use the liquid phase detection method to determine the contents of luteolin and isoquercetin in the composite extract powders obtained in Examples 1-6 and Comparative Examples 7-9 above. The specific test method is as follows:

[0073] Liquid phase detection method for luteolin: Chromatographic column YWG C18 ODS column (250 mm × 4.6 mm, 5 μm), mobile phase: 0.5% acetic acid: acetonitrile = 70:30, column temperature: T = 30 °C, detection wavelength 269 nm, liquid phase velocity 1 ml / min.

[0074] Liquid phase detection method for isoquercetin: Chromatographic column YWG C18 ODS column (250 mm × 4.6 mm, 5 μm), mobile phase: 0.5% acetic acid: acetonitrile = 73:27, column temperature: T = 30 °C, detection wavelength 351 nm, liquid phase velocity 1 ml / min.

[0075] The results are shown in Table 1.1 below:

[0076] Table 1.1 Results of the determination of the content of the composite extract powder

[0077]

[0078] From Table 1.1 above, it can be seen from Example 2 and Comparative Example 9, Example 4 and Comparative Examples 7 and 8 that by using the enzymatic hydrolysis method to enzymatically hydrolyze celery seeds and sophora flower buds, compared with the extraction method using ethanol, the contents of luteolin and isoquercitrin obtained are significantly increased, especially the content of isoquercitrin has a large increase. Under the same ratio of celery seeds and sophora flower buds, the content of luteolin obtained by the enzymatic hydrolysis extraction method is 2.7 times higher than that obtained by the ethanol extraction method, and the content of isoquercitrin is more than 22 times higher.

[0079] Test Example 2 Hypouricemic effect of the composite extract

[0080] 120 SD rats, weighing 180 - 220 g, with half males and half females, were adaptively fed for one week.

[0081] The rats were divided into a blank control group (8 rats) and a hyperuricemia group. The hyperuricemia group was given adenine 100 mg / (kg*d) + ethambutol hydrochloride 300 mg / (kg*d) to establish a model. After the model was successfully established, it was randomly divided into a model group and a drug administration group. The drug administration group was given 500 mg / kg of the composite extract powder obtained in the above examples and comparative examples, as well as the control drug allopurinol 24 mg / kg, luteolin monomer 2 mg / kg, isoquercitrin monomer 20 mg / kg, and luteolin monomer 2 mg / kg + isoquercitrin monomer 20 mg / kg (equivalent to the content and mass ratio of the two substances in Example 4) 4 h after modeling every day, and the gavage volume was 1 mL / 100 g. The blank control group and the model group were given an equal volume of normal saline.

[0082] At the end of the 3rd week of the experiment, after fasting for 12 h, abdominal aorta blood was taken, serum was separated by centrifugation, and serum uric acid (UA), serum creatinine (CRE), and blood urea nitrogen (BUN) were detected. The results are shown in Table 2.1 below:

[0083] Table 2.1 Effects of each group on UA, serum creatinine CRE, and blood urea nitrogen BUN in rats

[0084] Group UA (umol / L) CRE (umol / L) BUN (umol / L) Normal group 68.39±19.35 70.44±17.90 7.13±1.74 Model group 153.28±33.60 129.64±30.22 15.23±0.96 Allopurinol group 80.32±25.60 67.99±21.54 8.93±1.46 Example 1 70.87±33.02 75.64±24.47 9.09±1.44 Example 2 56.55±25.51 62.12±25.33 6.32±0.88 Example 3 52.98±21.10 59.32±19.79 6.73±1.73 Example 4 49.65±15.21 66.39±22.57 6.24±1.09 Example 5 60.35±29.07 70.31±19.02 6.98±1.25 Example 6 71.43±16.99 76.66±10.98 10.10±1.88 Comparative example 7 98.75±24.42 90.32±30.00 9.46±1.58 Comparative example 8 100.42±21.65 84.56±10.45 10.87±1.66 Comparative example 9 90.12±38.15 80.11±16.38 9.35±1.45 Luteolin group 129±18.71 100.43±18.09 12.54±3.18 Isoquercetin group 108±23.42 90.26±9.02 12.07±2.33 Luteolin + isoquercetin group 70.13±36.51 66.41±20.08 6.99±1.28

[0085] As can be seen from Table 2.1 above, the main indicators UA, CRE, and BUN of the composite extract powder provided in Examples 2-5 of the present invention are significantly improved in hyperuricemia rats, and the effect is better than that of the marketed drugs, Example 1, Example 6, and Comparative Examples 7-9; in addition, the combination of luteolin and isoquercetin alone can also significantly improve the main indicators UA, CRE, and BUN in hyperuricemia rats; the composite extract containing luteolin and isoquercetin obtained by the enzymatic hydrolysis method of Example 4 of the present invention has a better uric acid-lowering effect than the composition directly using the combination of luteolin and isoquercetin.

[0086] Test Example 3 Inhibitory effects of the composite extract on XOD and EP4

[0087] As a key enzyme in the catabolism of nucleic acids in the body, XOD can not only catalyze the formation of xanthine from hypoxanthine, but also catalyze the formation of uric acid from xanthine. EP4 is a key receptor for the downstream signals of the COX2-PGE2-Eps inflammatory pathway. Antagonizing EP4 can block the downstream signaling pathway of PGE2 to achieve anti-inflammatory and analgesic effects.

[0088] Select the composite extract powder of Examples 2-4 with good effects in Test Example 2, and measure the inhibitory effects of the composite extract powder on XOD and EP4 respectively.

[0089] Determination of XOD inhibitory activity: Take 0.60 mL of phosphate buffer (0.10 mol / L, pH = 8.50), 0.20 mL of xanthine solution (2.00 mmol / L), and 0.10 mL of sample solutions with different concentrations in a 1.50 mL centrifuge tube and vortex mix; then add 0.20 mL of xanthine oxidase solution (0.10 U / mL) and react in a water bath at 25 °C for 30 min; after completion, add 0.20 mL of HCl solution (1.00 mol / L) to terminate the reaction. Add 10 μL of reference substances and test samples with different concentrations to the control tube and the test sample tube respectively, and measure the absorbance at a wavelength of 290 nm. The half inhibitory concentration IC50 is calculated by the probit regression method using SPSS statistical software.

[0090] EP4 inhibitory activity assay: The inhibition of EP4 was determined by detecting the change in the concentration of cAMP in cells. Rat EP4 receptor was expressed in CHO cells, which were placed in MEM medium containing 10% fetal bovine serum, 50 units / mL penicillin, 50 μg / mL streptomycin, and 0.5 mg / mL G418, and cultured at 37 °C under 5% carbon dioxide. These cells were seeded into 96-well plates at a concentration of 50,000 per well and cultured overnight. On the day of measurement, the medium was first replaced with α-MEM containing 2 μM indomethacin and 0.5% BSA, and then replaced with α-MEM-ibmx buffer (α-MEM containing 2 μM indomethacin, 1 mM 3-isobutylmethylxanthine [IBMX], and 0.5% BSA). The test sample was added to the α-MEM-IBMX buffer, and the cells were incubated with 100 nM PGE2 at 37 °C for 30 minutes. The medium was removed, and the cells were lysed with 0.2% TritonX-100 / PBS. The cAMP level in the lysate was measured using a homogeneous time-resolved fluorescence assay kit. The IC50 value was estimated using sigmoid-Emax nonlinear regression analysis. The results are shown in Table 3.1 below:

[0091] Table 3.1 Inhibitory effects of composite extract powders of each group on XOD and EP4

[0092]

[0093]

[0094] The experimental results showed that the composite extract powders of Examples 2-4 had obvious inhibition on both XOD and EP4, thus playing a role in antioxidant, reducing uric acid, and alleviating inflammation.

[0095] Therapeutic effect of the composition of the composite extract and febuxostat on gouty arthritis in Test Example 4

[0096] Forty-eight SD rats, weighing 180-210 g, half male and half female, were adaptively fed for one week.

[0097] The rats were divided into a blank control group (8 rats) and a model group. The blank control group was intraperitoneally injected with 1 mL / 100 g of normal saline, and the model group was intraperitoneally injected with 500 mg / kg of hypoxanthine suspension at 1 mL / 100 g. After intraperitoneal injection, the rats were anesthetized with ether. In the model group, 0.1 mL of sodium urate suspension was injected into the joint cavity at a 45° direction on the dorsal side of the right hind ankle joint of the rats. After successful modeling, the rats were randomly divided into a model group and a drug administration group. The blank group was injected with an equal volume of normal saline.

[0098] The administration group was intragastrically administered with 500 mg / kg of the composite extract powder obtained in Examples 2-4 above + 0.5 mg / kg of febuxostat, and 1 mg / kg of the control drug febuxostat. The intragastric administration volume was 1 mL / 100 g. The blank control group and the model group were given an equal volume of normal saline, and intragastric administration was continued for 3 days.

[0099] Mark the right hind ankle joint of the rats to be tested. At 1 h, 24 h, 48 h, and 72 h after modeling, a paw swelling meter was used to measure the volume of the right hind ankle joint of each group of rats below the mark. The joint swelling degree (mL) = the ankle joint volume after modeling - the ankle joint volume before modeling. At 72 h after modeling, abdominal aorta blood was taken, serum was separated by centrifugation, and the levels of serum inflammatory factors IL-6 and TNF-α were detected. The results are shown in Tables 4.1 and 4.2 below.

[0100] Table 4.1 Effects of the composite extract powder and febuxostat composition on the joint swelling degree of rats

[0101]

[0102]

[0103] Table 4.2 Effects of the composite extract powder and febuxostat composition on the serum uric acid and inflammatory factors of rats

[0104] Group <![CDATA[IL-6(ρ / ng·L －1 )]]> <![CDATA[TNF-αρ / ng·L －1 )]]> Normal group 35.49±7.12 93.06±4.15 Model group 88.91±4.85 153.77±9.34 Example 1 + febuxostat 61.17±3.54 120.31±6.44 Example 2 + febuxostat 47.18±2.09 99.30±7.14 Example 3 + febuxostat 44.71±8.02 106.35±4.99 Example 4 + febuxostat 39.51±4.11 101.70±8.91 Example 6 + febuxostat 53.26±4.95 109.25±3.71 Febuxostat group 65.42±5.37 132.09±9.57 Celecoxib + febuxostat group 51.18±2.71 110.08±6.50

[0105] As can be seen from Tables 4.1 and 4.2, when the composite extract powder provided in Examples 2-4 of the present invention is used in combination with febuxostat, the dosage of febuxostat can be reduced, and the joint swelling of gouty arthritis during the acute attack of gout can be significantly alleviated, and inflammatory factors can be inhibited. Among them, the composite extract powder provided in Example 4 has a better effect of alleviating the joint swelling of gouty arthritis during the acute attack of gout and inhibiting inflammatory factors when combined with febuxostat.

[0106] The technical features of the above-described embodiments can be combined arbitrarily. For the sake of brevity of description, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, it should be considered to be within the scope described in this specification.

[0107] The above-described embodiments only represent several implementation manners of the present invention. Their descriptions are relatively specific and detailed, but they should not be construed as limiting the scope of the invention patent. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present invention, several modifications and improvements can still be made, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the present invention patent should be subject to the appended claims.

Claims

1. Use of a composition of luteolin and isoquercetin and febuxostat in the preparation of a medicament for preventing and / or treating hyperuricemia and gout, wherein, In the composition of luteolin and isoquercetin, the mass ratio of luteolin to isoquercetin is 1:5 to 100; the composition of luteolin and isoquercetin is a composite extract obtained by enzymatic extraction using celery seeds and sophora flower buds as raw materials; The enzymatic extraction method of the celery seeds and sophora flower buds includes the following steps: Crush the celery seeds and sophora flower buds respectively to obtain celery seed powder and sophora flower bud powder; Add the celery seed powder and sophora flower bud powder to a buffer solution; Perform enzymatic extraction at a temperature of 40°C to 60°C for 2h to 4h to obtain an extract; Inactivate the enzyme in the extract for 20min to 30min, separate to obtain a filtrate, and dry to obtain a composite extract powder; The enzyme is one or more of cellulase, rhamnosidase, naringinase, and pectinase; In the raw materials, the celery seeds are 30% to 65%, and the sophora flower buds are 35% - 70%; or in the raw materials, the celery seeds are 81% to 90%, and the sophora flower buds are 10% - 19%; The buffer solution is an aqueous solution of citric acid and sodium phosphate dodecahydrate with a pH value of 3.5 to 6.5, and the weight ratio of the total weight of the celery seed powder and sophora flower bud powder to the weight of the buffer solution is 1:10 to 40.

2. The application according to claim 1, characterized in that, In the composition of luteolin and isoquercetin, the mass ratio of luteolin to isoquercetin is 1:8 to 60.

3. The application according to claim 2, wherein In the composition of luteolin and isoquercetin, the mass ratio of luteolin to isoquercetin is 1:9 to 12.

4. The application according to claim 1, wherein The gout is gouty arthritis.

5. The application according to claim 1, characterized in that In the raw materials, the celery seeds are 35%, and the sophora flower buds are 65%; or in the raw materials, the celery seeds are 81%, and the sophora flower buds are 19%.

6. The application according to claim 1, characterized in that, The buffer solution is an aqueous solution of citric acid and sodium phosphate dodecahydrate with a pH value of 4, and the weight ratio of the total weight of the celery seed powder and sophora flower bud powder to the weight of the buffer solution is 1:

10.

7. The application according to claim 1, characterized in that, The enzyme is 0.5% - 10% of the total weight of the celery seeds and sophora flower buds.

8. A composition for preventing and / or treating hyperuricemia and gout, characterized in that, The active ingredients of the composition are the composition of luteolin and isoquercetin, and febuxostat. The active ingredients form the composition in a single form or a mixed form. Among them, the mass ratio of luteolin to isoquercetin is 1:5 to 100; the composition of luteolin and isoquercetin is a composite extract obtained by enzymatic extraction using celery seeds and sophora flower buds as raw materials; The enzymatic extraction method of the celery seeds and sophora flower buds includes the following steps: Crush the celery seeds and sophora flower buds respectively to obtain celery seed powder and sophora flower bud powder; Add the celery seed powder and sophora flower bud powder to a buffer solution; Perform enzymatic extraction at a temperature of 40°C to 60°C for 2h to 4h to obtain an extract; Inactivate the enzyme in the extract for 20min to 30min, separate to obtain a filtrate, and dry to obtain a composite extract powder; The enzyme is one or more of cellulase, rhamnosidase, naringinase, and pectinase; in the raw materials, the celery seeds are 30% to 65%, and the sophora flower buds are 35% - 70%; or in the raw materials, the celery seeds are 81% to 90%, and the sophora flower buds are 10% - 19%.

9. The composition according to claim 8, wherein In the composition of luteolin and isoquercetin, the mass ratio of luteolin to isoquercetin is 1:8 to 60.

10. The composition according to claim 9, wherein, In the composition of luteolin and isoquercetin, the mass ratio of luteolin to isoquercetin is 1:9 to 12.

Images