# US 11,298,390 B2 · Total flavonoids extract of Gynura formosana Kitam., preparation method therefor and use thereof for treating hyperuricemia > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,298,390 B2 - Document ID: US11298390B2 - Status: Granted - Kind: B2 - Application number: 16/821,314 - Filed: 2020-03-17 - Published: 2020-07-16 - Granted: 2022-04-12 - Earliest claimed: 2017-09-18 - Assignees: Zhangzhou Pien Tze Huang Pharmaceutical Co., Ltd. - Applicants: Zhangzhou Pien Tze Huang Pharmaceutical Co., Ltd. - Inventors: Huijun Yuan; Tingting Yin; Fei Hong; Juan Yu; Jinxiang Zeng; Shicong Wang - Web page: https://patentgrep.com/patent/US11298390B2 ## CPC - A61K 36/28: Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea - A23L 33/105: Plant extracts, their artificial duplicates or their derivatives - A61K 9/00: Medicinal preparations characterised by special physical form {(nuclear magnetic resonance contrast preparations or magnetic resonance imaging contrast preparations A61K49/18; preparations containing radioactive substances A61K51/12)} - A61K 31/7048: having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin {, digitoxin or digoxin} - A61P 19/06: Antigout agents, e.g. antihyperuricemic or uricosuric agents - B01D 15/08: Selective adsorption, e.g. chromatography - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - A23L: FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF (preservation thereof A23B) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - B01D: SEPARATION (separating solids from solids by wet methods B03B, B03D; by pneumatic jigs or tables B03B; by other dry methods B07; magnetic or electrostatic separation of solid materials from solid materials or fluids, separation by high-voltage electric fields B03C; centrifuges, vortex apparatus B04; presses per se for squeezing-out liquid from liquid-containing material B30B9/02; treatment of water C02F, e.g. softening by ion-exchange C02F1/42; {arrangements of air intake cleaners in gas turbine plants F02C7/05} ; arrangements or mounting of filters in air-conditioning, air-humidification or ventilation F24F13/28) ## Abstract Disclosed is a total flavonoids extract of Gynura formosana Kitam., containing 80%-85% of rutin by weight percent content. The results of pharmacological experiments show that the extract can lower the activity of xanthine oxidase in the liver of a hyperuricemia model mouse and reduce the synthesis of uric acid to a certain degree and has a certain uric acid-lowering effect, and can be used as a potential drug for treating hyperuricemia or gout. The method for preparing the total flavonoids extract of Gynura formosana Kitam. comprises: after an extraction step, selecting a complex enzyme composed of enzymes with a specific composition and a specific ratio for enzymolosis, and further carrying out the step of extracting and concentrating with macroporous resin and isolating and purifying with macroporous resin, so that the HPLC purity of rutin in the resulting total flavonoids extract of Gynura formosana Kitam. reaches 80%-85%. ## Claims ### Claim 1 A method for treating hyperuricemia or gout, comprising the step of administrating a total flavonoid extract from Gynura formosana Kitam. to a subject in need, wherein the total flavonoid extract from Gynura formosana Kitam., comprises, in weight percent, 80-85% of rutin and wherein the total flavonoid extract from Gynura formosana Kitam. is prepared by the steps of: (1) Extraction: extracting Gynura formosana Kitam. with an extraction solvent to obtain an extraction solution, and adjusting the extraction solution to a pH of 4-8 to obtain a reaction solution; (2) Enzymolysis: adding a complex enzyme into the reaction solution to carry out enzymolysis through a forced circular reaction at a temperature of 30° C. to 50° C. for 1 to 4 hours, then carrying out suction filtration, and collecting a filtrate; (3) Extraction and Concentration: extracting the filtrate by using a macroporous resin A to obtain an extracted solution, and concentrating the extracted solution to obtain a concentrated solution; and (4) Separation and Purification: centrifuging the concentrated solution, collecting a supernatant and carrying out elution by using a macroporous resin B, measuring absorbance at a wavelength of 510 nm, collecting eluate, concentrating and drying the eluate to obtain an extract. ### Claim 2 (depends on 1) The method of claim 1, wherein the complex enzyme used in the enzymolysis step consists of papain, cellulase and pectinase; and wherein a weight ratio of the complex enzyme to the Gynura formosana Kitam. is 1:5 to 1:3. ### Claim 3 (depends on 1) The method of claim 1, wherein a weight ratio of papain to cellulase to pectinase in the complex enzyme is (0.5-1.5):(2-5):(1-3). ### Claim 4 (depends on 1) The method of claim 1, wherein the macroporous resin A is one or more selected from the group consisting of AB-8, DM-130, HZ841, ZH-00, ZH-01, ZH-02, ZH-03, CAD-40, CAD-45 and BS-30; and the macroporous resin B is one or more selected from the group consisting of D-101, D-140, D-141, XAD-3, XAD-4, HP-20, HP-21, LD-605 and LSA-10. ### Claim 5 (depends on 1) The method of claim 1, wherein in the extraction step, the extraction solvent is water, and a weight ratio of Gynura formosana Kitam. to water is 1:(20-60). ### Claim 6 (depends on 1) The method of claim 1, wherein: in the separation and purification step, an ethanol aqueous solution with a volume concentration of 70-80% is adopted as an elution solvent, and the elution is performed at a rate of 3-15 m/h; and the concentrated solution comprises total flavonoid from Gynura formosana Kitam. at a concentration of 0.5 mg/mL. ### Claim 7 (depends on 2) The method of claim 2, wherein a weight ratio of papain to cellulase to pectinase in the complex enzyme is (0.5-1.5): (2-5):(1-3). ### Claim 8 (depends on 7) The method of claim 7, wherein the weight ratio of papain to cellulase to pectinase in the complex enzyme is 1:3:2. ### Claim 9 (depends on 2) The method of claim 2, wherein the macroporous resin A is one or more selected from the group consisting of AB-8, DM-130, HZ841, ZH-00, ZH-01, ZH-02, ZH-03, CAD-40, CAD-45 and BS-30; and the macroporous resin B is one or more selected from the group consisting of D-101, D-140, D-141, XAD-3, XAD-4, HP-20, HP-21, LD-605 and LSA-10. ### Claim 10 (depends on 2) The method of claim 2, wherein in the extraction step, the extraction solvent is water, and a weight ratio of Gynura formosana Kitam. to water is 1:(20-60). ### Claim 11 (depends on 2) The method of claim 2, wherein: in the separation and purification step, an ethanol aqueous solution with a volume concentration of 70-80% is adopted as an elution solvent, and the elution is performed at a rate of 3-15 m/h; and the concentrated solution comprises total flavonoid from Gynura formosana Kitam.at a concentration of 0.5 mg/mL. ### Claim 12 (depends on 3) The method of claim 3, wherein the weight ratio of papain to cellulase to pectinase in the complex enzyme is 1:3:2. ### Claim 13 (depends on 3) The method of claim 3, wherein the macroporous resin A is one or more selected from the group consisting of AB-8, DM-130, HZ841, ZH-00, ZH-01, ZH-02, ZH-03, CAD-40, CAD-45 and BS-30; and the macroporous resin B is one or more selected from the group consisting of D-101, D-140, D-141, XAD-3, XAD-4, HP-20, HP-21, LD-605 and LSA-10. ### Claim 14 (depends on 3) The method of claim 3, wherein in the extraction step, the extraction solvent is water, and a weight ratio of Gynura formosana Kitam. to water is 1:(20-60). ### Claim 15 (depends on 3) The method of claim 3, wherein: in the separation and purification step, an ethanol aqueous solution with a volume concentration of 70-80% is adopted as an elution solvent, and the elution is performed at a rate of 3-15 m/h; and the concentrated solution comprises total flavonoid from Gynura formosana Kitam.at a concentration of 0.5 mg/mL. ### Claim 16 A method for treating hyperuricemia or gout, comprising the step of administering to a subject in need a pharmaceutical preparation comprising a total flavonoid extract from Gynura formosana Kitam. as an active ingredient, wherein the total flavonoid extract from Gynura formosana Kitam., comprises, in weight percent, 80-85% of rutin, wherein the active ingredient is mixed with a conventional auxiliary material and prepared according to a conventional process into clinically acceptable forms selected from the group consisting of tablets, capsules, powders, mixtures, pills, granules, syrups, plasters, suppositories, aerosols, ointments and injections; and wherein the total flavonoid extract from Gynura formosana Kitam. is prepared by the steps of: (1) Extraction: extracting Gynura formosana Kitam. with an extraction solvent to obtain an extraction solution, and adjusting the extraction solution to a pH of 4-8 to obtain a reaction solution: (2) Enzymolysis: adding a complex enzyme into the reaction solution to carry out enzymolysis through a forced circular reaction at a temperature of 30° C. to 50° C. for 1 to 4 hours, then carrying out suction filtration, and collecting a filtrate; (3) Extraction and Concentration: extracting the filtrate by using a macroporous resin A to obtain an extracted solution, and concentrating the extracted solution to obtain a concentrated solution; and (4) Separation and Purification: centrifuging the concentrated solution, collecting a supernatant and carrying out elution by using a macroporous resin B, measuring absorbance at a wavelength of 510 nm, collecting eluate, concentrating and drying the eluate to obtain an extract.