Flavonoid active site of Abelmoschus manihot flower, its preparation method and use

Abstract

The present invention relates to the flavonoid effective site of Abelmoschus manii flower and its preparation method and use, which belongs to the field of Chinese medicine pharmaceuticals.The present invention prepares high-purity Abelmoschus manii flower flavonoid effective site, and finds that it has better nephropathy treatment activity, especially has remarkable treatment effect on diabetic nephropathy, zebra lupus nephropathy, or contrast agent-induced nephropathy, ophthalmological disease.At the same time, the flavonoid effective site of Abelmoschus manii flower described in the present invention has good safety.

Description

Technical Field

[0003] , ,

[0001] The present invention relates to the field of traditional Chinese medicine manufacturing, specifically to the flavonoid active site of Abelmoschus manihot flower and its use related matters.

Background Art

[0002] Medicinal Abelmoschus manihot flower is the dried corolla of the Abelmoschus manihot (L.) Medic plant of the Malvaceae family. It tastes sweet and cold, belongs to the meridians of the kidney and bladder, has the effects of removing dampness and heat, relieving swelling, and removing toxins, and is used for carbuncles, gangrene, and burns caused by water or fire. This medicine was first recorded in the "Jiayou Bencao", and it was recorded in the "Jiayou Bencao" as follows: Abelmoschus manihot flower is used for external application to treat those with malignant ulcers and abscesses that do not heal for a long time. It was recorded in the "Compendium of Materia Medica" as follows: Abelmoschus manihot flower, its flower has a sweet fragrance, is cold, slippery, non-toxic, and can be used for external application to treat those with malignant ulcers and abscesses that do not heal for a long time, and is the main therapeutic drug for patients with chronic ulcers.

[0003] Flavonoid compounds are not only one of the main components of the Abelmoskasmanihot flower, but also one of its pharmacologically active ingredients. Abelmoskasmanihot capsules have been on the market for many years, and their main active ingredient is an extract of the Abelmoskasmanihot flower. Studies have confirmed that the flavonoid components in the Abelmoskasmanihot flower extract have a remarkable therapeutic effect on sores and pus, and also have a remarkable protective effect on damage to the heart, brain, and tissues caused by ischemia. Furthermore, numerous studies have reported that the total flavonoids of the Abelmoskasmanihot flower may exhibit superior effects in areas such as anti-inflammatory, antipyretic, analgesic, protection against cardiocerebral ischemic damage, hypoglycemic, and antiviral properties. For example, Chinese Patent Application No. 201210082553.7 discloses a total flavonoid extract of Abelmoscus manihot flower, containing quercetin-3'-glucoside, quercetin-3'-glucoside, and isoquercitrin in a weight ratio of (11-16):(2.5-6):(4-6.5), and is used in the preparation of therapeutic drugs for nephropathy. Chinese Patent Application No. 200610097615.6 discloses a total flavonoid extract of Abelmoscus manihot flower, with a total flavonoid content of 50-90% by weight, and the flavonoid component being quercetin-3-robinobioside 1.0-5.0%. hyperoside The total flavonoid extract of Abelmoscus manihot flower contains 8-24.0% of isoquercitrin, 7.0-20.0% of quercetin-3'-glucoside, 5.0-15.0% of quercetin-3'-glucoside, 3.0-10.0% of quercetin-3'-glucoside, 0.5-5.0% of myricetin, 0.5-5.0% of quercetin, 2.0-8.0% of quercetin, and several other flavonoid components. The total flavonoid extract of Abelmoscus manihot flower is used in the preparation of nephropathy treatment drugs, and the qualitative and quantitative determination of the flavonoid components is clear, ensuring therapeutic effects.

[0004] Despite the various therapeutic effects of bioflavonoids, there are numerous types of flavonoids, which can currently be classified into seven main subtypes, and different flavonoid compositions result in different therapeutic effects. For example, while bioflavonoids have been reported in the literature to be effective in preventing and treating diabetic retinopathy, compounds with five hydroxyl groups, such as quercetin, negatively affect ocular blood flow. However, dehydrogenating flavonoids to flavanones significantly improves ocular blood flow. Furthermore, compounds that increase blood flow significantly increase retinal function recovery after ischemic injury. How to develop and effectively utilize such compounds to enhance their effects on treating eye diseases such as macular degeneration, visual fatigue, and cataracts, or on diabetic retinopathy, is an important research challenge.

[0005] Choroidal neovascularization (CNV) is seen in many fundus diseases, including age-related macular degeneration (AMD), central serous exudative choroidal retinitis, and macular degeneration due to high myopia. Among these, CNV secondary to AMD is the most common and is one of the main causes of irreversible visual impairment in the elderly. Poor blood flow is also a factor that affects visual fatigue, and promoting blood flow to the eye and acting on the smooth muscles of the eye contributes to recovering from visual fatigue.

[0006] Proximal renal tubular glucose reabsorption is mediated by sodium-glucose cotransporter proteins (SGLT)1 and SGLT2, with approximately 90% of glucose reabsorption mediated by SGLT2 and the remaining 10% by SGLT1. SGLT2 inhibitors selectively bind to SGLT2 receptors, thereby inhibiting renal tubular glucose reabsorption, lowering the renal glucose threshold, increasing urinary glucose excretion, and achieving significant blood glucose reduction. Recent clinical studies have shown that SGLT2 inhibitors have renoprotective effects against chronic kidney disease (CKD) not caused by diabetes. SGLT2 inhibitors can reduce the expression of collagen and fibronectin by decreasing the expression of TGFβ-1, PAI1, STAT1, and MMP7, and by inhibiting the AGEs-RAGE axis, thereby reducing the accumulation of extracellular matrix and mitigating the progression of DN renal fibrosis. SGLT2 inhibitors modulate tubular bulb feedback, reducing the process of proteinuria and, in addition to their anti-inflammatory and anti-fibrotic effects, can also reduce endogenous uric acid (SUA) in diabetic patients. For example, compared to a control group, SGLT2 inhibitors (empagliflozin, cargliflozin, dagliflozin, togliflozin, lugliflozin, igliflozin) significantly reduce serum uric acid levels. SGLT2 inhibitors may be beneficial in the prevention and treatment of heart failure due to reduced plasma volume, reduced pre- and post-load, improved myocardial energy metabolism, and improved myocardial remodeling.

[0007] Methods for extracting Abelmoskasmanihot flowers reported in the literature include ethanol reflux extraction, ultrasonic extraction, and room temperature immersion extraction. However, the components of Abelmoskasmanihot flower extracts are complex, the content of active ingredients varies greatly, the extraction transfer rate is low, the energy consumption of extraction is high, and the amount of solvent used is also high. Therefore, there is a need to explore Abelmoskasmanihot flower therapeutic drugs and extraction methods that can be combined to treat various diseases simultaneously, such as nephropathy, especially diabetic nephropathy, and ophthalmic diseases.

[0008] The applicant compiled statistics on adverse drug reactions of Abelmoscus capsules and found 194 cases of adverse reactions in the past year. According to the statistics on drug adverse reaction occurrence, nausea, itching, rash, vomiting, diarrhea, epigastric bloating, and discomfort are all known adverse reactions, with more than 10 occurrences. Recently emerged ADR symptoms include dizziness, headache, and loss of appetite. Furthermore, one serious adverse reaction reported was hepatocyte damage, which may suggest hepatocyte damage caused by Abelmoscus capsules. Therefore, developing a new flavonoid extract from Abelmoscus manihot flowers to mitigate side effects and prepare a drug for disease treatment has clinical significance. [Overview of the project] [Problems that the invention aims to solve]

[0009] In view of the above drawbacks, the present invention provides the flavonoid active site of Abelmoscus manihot flower and its uses. The present invention provides a high-purity flavonoid active site of Abelmoscus manihot flower prepared by the extract-binding resin method, which exhibits good therapeutic activity for nephropathy, such as diabetic nephropathy, contrast-induced nephropathy, or lupus erythematosus nephritis, and also has an effect in treating ophthalmic diseases. At the same time, the flavonoid active site, extract, or composition of Abelmoscus manihot flower described in the present invention has few side effects and can provide patients with a better therapeutic drug. [Means for solving the problem]

[0010] To solve the above technical problems, the present invention provides the following technical solutions.

[0011] In a first embodiment, the present invention provides a flavonoid active site of Abelmoscus manihot flower containing flavonoid components in the following mass ratios: Goshipechin -8-O-β-D-glucuronide: hyperoside The mass ratio of isoquercitrin, myricetin, and quercetin-3'-O-glucoside is 10:3.0~20:4.0~18:1.9~6.0:6.0~20.

[0012] specifically, Goshipechin -8-O-β-D-glucuronide: hyperoside The mass ratio of isoquercitrin:myricetin:quercetin-3'-O-glucoside is 10:4.0~17:4.50~16:1.9~5.5:7.0~16, preferably 10:6.0~15:5.0~13:2.0~5.0:8.0~14, and further containing quercetin, where, Goshipechin -8-O-β-D-glucuronide:quercetin is in a ratio of 10:1.0 to 10.0, preferably 10:1.0 to 6.0, preferably 10:1.5 to 5.0, and further contains rutin, where, Goshipechin -8-O-β-D-glucuronide:Rutin is in a ratio of 10:0.05~0.6, preferably 10:0.1~0.4, more preferably 10:0.1~0.3, and even more preferably contains quercetin-3-O-robinobioside, where, Goshipechin -8-O-β-D-glucuronide:quercetin-3-O-robinobioside has a ratio of 10:0.1~2.5 and 10:0.1~0.9, preferably 10:0.15~0.5.

[0013] In a second aspect, the present invention provides a flavonoid active site of Abelmoscus manihot flower containing flavonoid components in the following mass ratio: isoquercitrin:quercetin:quercetin-3'-O-glucoside:myricetin: Goshipechin -8-O-β-D-glucuronide: hyperoside The mass ratios are 0.8-1.2:0.05-1.6:0.2-4.6:0.05-1.2:0.2-3.5:0.25-3.6.

[0014] Specifically, the flavonoid active portion of the Abelmoscus manihot flower contains flavonoid components in the following mass ratio: isoquercitrin:quercetin:quercetin-3'-O-glucoside:myricetin: Goshipechin -8-O-β-D-glucuronide: hyperosideThe mass ratios are 0.8~1.2:0.1~1.2:0.4~3.1:0.10~0.9:0.4~3.1:0.5~3.0, and further 0.8~1.2:0.16~1.0:0.6~2.4:0.14~0.75:0.6~2.5:0.6~2.4, and further 0.8~1.2:0.2~0.8:0.8~2 1:0.18~0.6:0.8~2.0:0.7~2.0, further 0.8~1.2:0.2~0.7:0.8~1.6:0.2~0.5:0.8~1.8:0.8~1.4, further 0.8~1.2:0.2~0.6:1.0~1.2:0.2~0.4:0.9~1.7:1.0~1.3.

[0015] Specifically, the flavonoid active parts of the Abelmoscus manihot flower are in the following mass ratio: isoquercitrin:quercetin:quercetin-3'-O-glucoside:myricetin: Goshipechin -8-O-β-D-glucuronide: hyperoside The mass ratio of the flavonoid components is 0.8~1.2:0.09~1.6:0.2~2.0:0.05~1.2:0.5~3.0:0.25~3.6.

[0016] Specifically, the flavonoid active parts of the Abelmoscus manihot flower are in the following mass ratio: isoquercitrin:quercetin:quercetin-3'-O-glucoside:myricetin: Goshipechin -8-O-β-D-glucuronide: hyperoside The mass ratio of these components is 0.8~1.2:0.12~0.7:0.8~1.6:0.2~0.5:0.85~1.7:1.0~1.4, and they contain flavonoid components.

[0017] More specifically, the mass ratio of isoquercitrin is 1.2 or 1.0.

[0018] More specifically, the flavonoid active site of Abelmoschus manihot flower further contains rutin, and the mass ratio of isoquercitrin to rutin is 1:0.001 to 0.08, preferably 1:0.005 to 0.06, preferably 1:0.006 to 0.05, preferably 1:0.007 to 0.04, preferably 1:0.009 to 0.04, or preferably 1:0.008 to 0.03, preferably 1:0.009 to 0.03. Furthermore, it contains quercetin-3-O-rutinoside, and the mass ratio of isoquercitrin to quercetin-3-O-rutinoside is 10:0.1 to 2.5, 10:0.1 to 0.9, preferably 10:0.15 to 0.5.

[0019] More specifically, the flavonoid active site of Abelmoschus manihot flower contains quercetin, quercetin-3'-O-glucoside, myricetin, Goshipechin -8-O-β-D-glucuronide, isoquercitrin and hyperoside The total content of which is 55% or more, preferably 60% or more, preferably 65 - 85%, preferably 69 - 82%. Furthermore, the content of quercetin-3'-o-glucoside is 12.1 - 25%. Additionally, the content of quercetin-3'-o-glucoside is 12.6 - 23% or 13 - 20%, 13 - 20%, or 14 - 25%. Or furthermore, the content of quercetin exceeds 0.7%, further exceeds 1.0%, and is further 1.0% - 10%, further 1.5% - 5%. Or additionally, Goshipechin -8-O-β-D-glucuronide is 8.5 - 30%, further 12% - 23%.

[0020] More specifically, the active site contains quercetin, quercetin-3'-O-glucoside, myricetin, Goshipechin -8-O-β-D-glucuronide, isoquercitrin, rutin and hyperosideThe total content is 55% or more, preferably 60% or more, preferably 65 - 85% or 66 - 84%, preferably 69 - 82%, or 69 - 90%. Furthermore, the quercetin - 3'-o - glucoside content is 12.1 - 25%, and furthermore, the quercetin - 3'-o - glucoside content is 12.6 - 23% or 13 - 20%.

[0021] In a third aspect, the present invention provides a plant extract containing a flavonoid component containing components with the following mass contents: hyperoside 10 - 25%, isoquercitrin 8 - 19%, Goshipechin -8 - O - β - D - glucuronide 5 - 30%, quercetin - 3'-o - glucoside 12.1 - 25%, and furthermore hyperoside 12 - 23%, isoquercitrin 10 - 17%, Goshipechin -8 - O - β - D - glucuronide 8.5 - 25%, quercetin - 3'-o - glucoside 12.6 - 23%, and furthermore hyperoside 13 - 22%, isoquercitrin 11 - 17%, Goshipechin -8 - O - β - D - glucuronide 12 - 21%, quercetin - 3'-o - glucoside 13 - 20%.

[0022] Specifically, the extract further contains 2 - 10% quercetin, preferably contains 2.5 - 9% quercetin, preferably contains 3 - 8.5% quercetin, further contains 1.5% - 5%, and furthermore, contains 2 - 11% myricetin, preferably contains 3 - 7% myricetin, preferably contains 3 - 5% myricetin, and furthermore, contains 0.08 - 2.5% quercetin - 3 - O - robinoside, preferably 0.1 - 1.5%, further 0.1 - 0.9%, more preferably 0.1 - 0.5%. Furthermore, the plant is Abelmoschus manihot or okra flower, which may be the whole flower or the corolla, preferably Abelmoschus manihot flower.

[0023] In a fourth aspect, the present invention provides an Abelmoschus manihot flower extract containing a flavonoid component containing components with the following mass contents: hyperoside 8-26%, isoquercitrin 12.0-19.8%, Goshipechin -8-O-β-D-glucuronide 8.5-30%, myricetin 3.0-4.9% or 5.1-6.0%, quercetin-3'-o-glucoside 14-25%, quercetin 1.6-4.9%, and further hyperoside 11-22%, isoquercitrin 12.0-17%, Goshipechin The composition is 12-23% -8-O-β-D-glucuronide, 1.6-4.9% or 5.1-9.0% myricetin, 13-22% quercetin-3'-o-glucoside, and 1.4-8% or 1.4-7.8% quercetin.

[0024] Specifically, the extract further contains rutin in a mass content of 0.01 to 1.0%, further 0.05 to 0.8%, further 0.09 to 0.8%, and further 0.1 to 0.6%, and further contains quercetin-3-O-robinobioside in a mass content of 0.08 to 2.5%, preferably 0.1 to 1.5%, further 0.1 to 0.9%, and even more preferably 0.1 to 0.5%.

[0025] Specifically, the mass content of flavonoid components in the Abelmoscus manihot flower extract is 55% or more, preferably 60% or more, preferably 65-85% or 66-84%, and preferably 69-82% or 69-90%.

[0026] In a fifth aspect, the present invention provides a composition containing flavonoid components in the following mass ratio: isoquercitrin:quercetin:quercetin-3'-O-glucoside:myricetin: Goshipechin -8-O-β-D-glucuronide: hyperosideThe mass ratios are 1:0.05~1.6:0.2~4.6:0.05~1.2:0.2~3.5:0.25~3.6, further 1:0.1~1.2:0.4~3.1:0.10~0.9:0.4~3.1:0.5~3.0, and further 1:0.16~1.0:0.6~2.4:0.14~0.75:0.6~2.5:0.6 ~2.4, further 1:0.2~0.8:0.8~2.1:0.18~0.6:0.8~2.0:0.7~2.0, further 1:0.2~0.7:0.8~1.6:0.2~0.5:0.8~1.8:0.8~1.4, further 1:0.2~0.6:1.0~1.2:0.2~0.4:0.9~1.7:1.0~1.3 And then there are 1.2:0.05~1.6:0.2~4.6:0.05~1.2:0.2~3.5:0.25~3.6, and furthermore 1:0.1~1.2:0.4~3.1:0.10~0.9:0.4~3.1:0.5~3.0, and furthermore 1:0.16~1.0:0.6~2.4:0.14~0.75:0.6~2.5:0.6~2. 4. Furthermore, 1:0.2~0.8:0.8~2.1:0.18~0.6:0.8~2.0:0.7~2.0, further 1:0.2~0.7:0.8~1.6:0.2~0.5:0.8~1.8:0.8~1.4, and further 1:0.2~0.6:1.0~1.2:0.2~0.4:0.9~1.7:1.0~1.3.

[0027] Specifically, the composition further contains rutin, the mass ratio of isoquercitrin to rutin is 1:0.001~0.08, preferably 1:0.005~0.06, preferably 1:0.006~0.05, preferably 1:0.007~0.04, preferably 1:0.008~0.03, preferably 1:0.009~0.03, the quercetin-3'-o-glucoside content in the flavonoid component is 12.1~25%, the quercetin-3'-o-glucoside content is 12.6~23% or 13~20%, and the composition further contains 0.08~2.5% quercetin-3-O-robinobioside, preferably 0.1~1.5%, more preferably 0.1~0.9%, and even more preferably 0.1~0.5%.

[0028] More specifically, the composition contains quercetin, quercetin-3'-O-glucoside, myricetin, Goshipechin -8-O-β-D-glucuronide, isoquercitrin, rutin and hyperoside The total content of is 55% or more, preferably 60% or more, preferably 65-85%, and preferably 69-82%.

[0029] In a sixth embodiment, the present invention provides a pharmaceutical composition comprising the above-mentioned active site, the above-mentioned Abelmoscus manihot extract, or the above composition, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

[0030] Specifically, the pharmaceutically acceptable carrier comprises a solvent, emulsifier, disintegrant, filler, solubilizer, antioxidant, pH adjuster, osmotic pressure adjuster, bacteriostatic agent, diluent, lubricant, binder and / or film-forming agent, wherein the lubricant does not contain magnesium stearate.

[0031] Specifically, the dosage forms of the pharmaceutical composition include injections, tablets, suppositories, ointments, gels, pills, granules, capsules, or compounded preparations, suspensions, and powders.

[0032] In a seventh embodiment, the present invention provides uses of the above-mentioned active site, the above-mentioned Abelmoscus manihot extract, the above-mentioned composition, or the above-mentioned pharmaceutical composition in the preparation of nephropathy drugs, wherein the nephropathy is preferably diabetic nephropathy or diabetic nephropathy or nephritis with renal fibrosis.

[0033] In the eighth embodiment, the present invention provides uses of the above-mentioned active site, the above-mentioned Abelmoscus manihot extract, the above-mentioned composition, or the above-mentioned pharmaceutical composition in the preparation of ophthalmic disease drugs, the ophthalmic disease being preferably macular degeneration, visual fatigue or cataract, diabetic retinopathy, age-related macular degeneration, central serous exudative chorioretinitis, or macular degeneration due to high myopia.

[0034] In a ninth embodiment, the present invention provides uses of the above-mentioned active site, the above-mentioned Abelmoscus manihot extract, the above-mentioned composition, or the above-mentioned pharmaceutical composition in the preparation of drugs for lupus erythematosus nephritis, contrast-induced renal impairment, pulmonary fibrosis, heart failure, hypouricemia, or SGLT2 inhibitors.

[0035] In a tenth embodiment, the present invention provides a method for preparing the above-mentioned active site or the above-mentioned Abelmoscus manihot extract, the method comprising: (1) extracting the Abelmoscus manihot flower or the medicinal site of Abelmoscus manihot with ethanol to prepare an extract; and (2) concentrating the extract and then extracting (Extraction) The process includes the steps of (1) preparing an extract solution, (2) removing the solvent from the extract solution and then eluting it with a macroporous resin to prepare the flavonoid active parts or extract of Abelmoscus manihot flower, preferably by percolation.

[0036] Specifically, the amount of ethanol used in step (1) is 10 to 25 times the amount of Abelmoskasmanihot flower or the medicinal part of Abelmoskasmanihot, and the ethanol is a 60 to 95% ethanol solution; the extractant for the extract in step (2) is n-butanol, petroleum ether, or ethyl acetate; the extracting method is a continuous countercurrent extract; the liquid-material ratio of the extract is 0.8 to 4:1; the number of extracting stages of the extract is 1 to 5; and the macroporous resin type in step (3) is D101, HPD100, or AB-8.

[0037] Specifically, step (2) further includes subjecting the extract to activated carbon adsorption, alcohol precipitation, or acid precipitation treatment before extraction.

[0038] Specifically, the elution process for the macroporous resin in step (3) is as follows: the diameter-to-height ratio of the macroporous resin is 1:4 to 1:9, the loading buffer concentration is 0.10 to 0.30 g raw material / mL, the loading buffer volume is 4 to 12 BV, loading is performed at a flow rate of 1 to 4 BV / h, impurities are removed using 4 to 8 BV of pure water and 1 to 5 BV of 3 to 15% ethanol at a flow rate of 0.5 to 4 BV / h, and elution is performed using 2 to 8 BV of 50 to 80% ethanol at a flow rate of 1 to 5 BV / h.

[0039] In an eleventh embodiment, the present invention provides a method for preparing the above-mentioned active site or the above-mentioned Abelmoskasmanihot extract, the preparation method comprising: (1) extracting the Abelmoskasmanihot flower or the medicinal site of Abelmoskasmanihot with ethanol to prepare an extract; (2) adding a clarifying agent to the extract, treating it in a water bath, filtering it to remove the supernatant; and (3) eluting the supernatant with a polyamide resin to prepare the flavonoid active site or extract of Abelmoskasmanihot flower, wherein the extraction method is preferably percolation.

[0040] Specifically, the amount of ethanol used in step (1) is 10 to 25 times the amount of Abelmoskasmanihot flower or Abelmoskasmanihot medicinal part, and the ethanol is a 60 to 95% ethanol solution; the water bath temperature in step (2) is 50 to 70°C, the water bath time is 30 to 90 minutes; the resin diameter to height ratio in step (3) is 1:4 to 1:9; the loading buffer concentration is 0.10 to 0.60 g raw material / mL; the loading buffer volume is 4 to 12 BV; and elution is performed using 4 to 8 BV of pure water and 4 to 8 BV of 60 to 95% ethanol.

[0041] In a twelfth embodiment, the present invention provides a method for preparing the above-mentioned active site or the above-mentioned Abelmoskasmanihot extract, the preparation method comprising: (1) extracting the Abelmoskasmanihot flower or the medicinal site of Abelmoskasmanihot with ethanol to prepare an extract; (2) adjusting the pH of the extract to 2.0 to 3.0, refrigerating it, filtering it to remove the precipitate, and adding water to dissolve it; and (3) the dissolved solution Extraction The process includes the steps of (4) preparing an extract solution, (5) removing the solvent from the extract solution and then eluting it with a polyamide resin to prepare the flavonoid active part or extract of Abelmoscus manihot flower, preferably by reflux.

[0042] Specifically, the amount of ethanol used in step (1) is 10 to 25 times the amount of Abelmoskasmanihot flower or Abelmoskasmanihot medicinal part, and the ethanol is a 60 to 95% ethanol solution; the pH adjuster in step (2) is hydrochloric acid, sulfuric acid, acetic acid, phosphoric acid, citric acid, tartaric acid, or maleic acid; the extractant for the extract in step (3) is n-butanol, petroleum ether, or ethyl acetate; the extraction method is a continuous countercurrent extract; the liquid-material ratio of the extract is 0.8 to 4:1; the number of extraction stages of the extract is 1 to 5; the diameter-to-height ratio of the resin in step (4) is 1:4 to 1:9; the loading buffer concentration is 0.10 to 0.60 g raw material / mL; the loading buffer volume is 4 to 12 BV; and elution is performed using 4 to 8 BV of pure water and 4 to 8 BV of 60 to 95% ethanol.

[0043] The flavonoid active part of the Abelmoscus manihot flower described in the present invention is hyperoside Rutin, isoquercitrin, Goshipechin In addition to detecting the seven components -8-O-β-D-glucuronide (hibifolin), myricetin, quercetin-3'-o-glucoside, and quercetin, during the detection process, the solid content sample prepared in the examples of the present invention is also converted to a content equivalent to the peak area proportional to the quercetin-3-O-robinobioside content, using the content conversion formula = hyperoside Control product peak area / hyperoside The control sample concentration*, correction factor*, quercetin-3-O-robinobioside peak area*, constant volume / weighed volume (after water removal), content is 2.0% or less, more preferably 0.9% or less, and more preferably 0.1% to 0.7%. The total content of the detected multiple flavonoids is 70% or more, more preferably 75% or more, and more preferably 80% to 90%. [Effects of the Invention]

[0044] The present invention has the following effects.

[0045] (1) The flavonoid active site or extract of Abelmoscus manihot flower described in the present invention not only has the effect of treating various nephropathy, such as diabetic nephropathy, contrast-induced nephropathy, or lupus erythematosus nephritis, but also has the effect of treating ophthalmic diseases, especially various diseases related to impaired blood flow and impaired ocular blood vessels, such as age-related macular degeneration, central serous exudative chorioretinitis, and macular degeneration due to high myopia, and in particular the flavonoid active site or extract of Abelmoscus manihot flower described in the present invention also has the effect of treating pulmonary fibrosis.

[0046] (2) The preparation method described in the present invention is simple in its process, has a short production cycle, mild processing conditions, low energy consumption, and good separation of the seven flavonoids of Abelmoscus manihot flower. The raw materials and reagents used are available from a wide range of sources, the cost is low, and industrialized large-scale production is easily realized.

[0047] (3) The flavonoid active part or extract or composition of Abelmoscus manihot flower described in the present invention has fewer side effects and provides a better therapeutic means for patients. [Brief explanation of the drawing]

[0048] [Figure 1]This graph shows the results of pathological changes detected when rats were repeatedly administered different doses over a 3-month period. A represents the normal group, B represents the Example 9.2 sample group, C represents the Abelmosque capsule group, and D represents the Example 2-A4 group. [Figure 2] This graph shows the pathological results for the DKD, HK, and HT groups. [Figure 3] This graph shows the expression detection results for SLC5A2 mRNA in the DKD, HK, and HT groups. [Modes for carrying out the invention]

[0049] The present invention will be described in more detail below in relation to specific examples, but the following examples are not intended to limit the present invention and are used solely for the purpose of illustrating the present invention. Unless otherwise specified, the experimental methods used in the following examples are usually carried out under conventional conditions, and unless otherwise specified, the materials, reagents, etc. used in the following examples are all commercially available.

[0050] The Abelmoscus manihot flowers in the following examples are the dried corollas of Abelmoscus manihot, and unless otherwise specified, they are Abelmoscus manihot flowers from the same batch.

[0051] The content of seven components was measured in the flavonoid active parts of the Abelmoscus manihot flowers prepared in each example. These seven components are: hyperoside Rutin, isoquercitrin, Goshipechin The components are -8-O-β-D-glucuronide, myricetin, quercetin-3'-o-glucoside, and quercetin. During the preparation of the active site samples, the content of each component was also measured in the solution of the active site before drying, and slight changes in the content of the samples before and after drying were observed. The detection method was the UPLC method reported in "Measurement of seven components of Abelmoscus manihot flower by one-test-multiple evaluation method" (Journal of Drug Analysis, 2013, No. 12, 2082-2087), which measured the content of each component.

[0052] The liquid-to-material ratio of the aforementioned extract (i.e., extractant:extract) is expressed as a volume ratio.

[0053] Example 1. Preparation of flavonoid-active parts of Abelmoscus manihot flowers 100g of Abelmoskasmanihot flowers were ground into a coarse powder and extracted using percolation with 15 times 60% ethanol to obtain an Abelmoskasmanihot flower extract. Ethanol was removed from the Abelmoskasmanihot flower extract, diluted with water, adsorbed with activated carbon, and then continuously extracted with n-butanol in a countercurrent manner. The liquid-to-material ratio of the extract (i.e., extractant:extract) was 1.5:1, the loading rate of the extract was 40 mL / min, the rotation speed of the extractor was 40 Hz, and the number of extract stages was 2. After recovering the solvent by reducing the pressure of the extract solution at 40°C, it was treated with D101 macroporous resin. The processing steps were as follows: The diameter-to-height ratio of the macroporous resin was 1:6, the loading buffer (aqueous solution) concentration was 0.15 g raw material / mL, the loading buffer volume was 7 BV, loading was performed at a flow rate of 2 BV / h, decontamination was sequentially performed at a flow rate of 2 BV / h using 6 BV of pure water and 3 BV of 5% ethanol, elution was performed at a flow rate of 2 BV / h using 4 BV of 60% ethanol to obtain an eluate, and then the eluate was reduced in pressure at 50°C to recover the ethanol and obtain a flavonoid extract of Abelmoscus manihot flower. The content was measured, and the content of the seven components was adjusted by the accession method according to the content measurement results. The content of each component is shown in the table below. The total solid content of the flavonoid active part of Abelmoscus manihot flower was 2.23 g, and the seven components totaled 1769 mg, accounting for 79.32% of the total solid content.

[0054] [Table 1]

[0055] Example 2. Preparation of flavonoid-active parts of Abelmoscus manihot flowers Example 2-1 100g of Abelmoskasmanihot flowers were ground into a coarse powder and extracted using percolation with 18 times 70% ethanol to obtain an Abelmoskasmanihot flower extract. The ethanol was removed from the Abelmoskasmanihot flower extract, diluted with water, and extracted using continuous countercurrent with ethyl acetate. The liquid-to-material ratio of the extract (i.e., extractant:extract) was 2.5:1, the loading rate of the extract was 80 mL / min, the rotation speed of the extractor was 40 Hz, and the number of extract stages was 4. After recovering the solvent by reducing the pressure of the extract solution at 40°C, it was treated with D101 macroporous resin. The following conditions were met: the diameter-to-height ratio of the macroporous resin was 1:5, the loading buffer concentration was 0.15 g raw material / mL, the loading buffer volume was 6 BV, loading was performed at a flow rate of 2 BV / h, impurities were removed at a flow rate of 2 BV / h using 6 BV of pure water and 3 BV of 5% ethanol, and elution was obtained by eluting at a flow rate of 1.5 BV / h using 4 BV of 60% ethanol. The eluate was then reduced under reduced pressure at 60°C to recover the ethanol, and an Abelmoscus manihot flower flavonoid extract was obtained. The content was measured, and the content of the seven components was adjusted by the accession method according to the content measurement results. The content of each component is shown in the table below. The total solid content of the flavonoid active site of the Abelmoscus manihot flower is 2.99. The content of quercetin-3-O-robinobioside is 0.28%.

[0056] [Table 2-1]

[0057] Example 2-A. Preparation of flavonoid-active parts of Abelmoscus manihot flowers Following the method of Example 2 described above, Abelmoscus manihot flowers were magnified 10-fold, and the flavonoid active sites of two other batches of Abelmoscus manihot flowers were prepared to obtain samples of the measurement data for sample content, as shown in Table 2-2. The quercetin-3-O-robinobioside content was 0.20-0.50%.

[0058] [Table 2-2]

[0059] Here, 2-A1 contains 0.24% of the content, converted to peak area, and the ratio relationship of each component is shown in Table 2-3 below.

[0060] [Table 2-3]

[0061] Example 2-A3 hyperoside Isoquercitrin, hibifolin, myricetin, quercetin-3'-o-glucoside, quercetin, and quercetin 3-O-robinobioside were taken as raw materials, and the components were mixed according to the ratio of Example 2-A1 (rutin was replaced with quercetin 3-O-robinobioside, and the ratio was adjusted to 0.02) to prepare a total flavonoid composition.

[0062] Example 2-A4 hyperoside Isoquercitrin, hibifolin, myricetin, quercetin-3'-o-glucoside, quercetin, and quercetin 3-O-robinobioside were selected as raw materials, and the components were mixed according to the ratio of Example 2-A1 (rutin was replaced with quercetin 3-O-robinobioside, and the ratio was adjusted to 0.05) to prepare a total flavonoid composition.

[0063] Example 2-B To prepare the active ingredient of Abelmoscus manihot flower that satisfies the requirement of mass uniformity, the relationship between the content of the drug and the extract of the aforementioned components was investigated from the content results of each component of the extract of multiple batches of Example 2-1. Three different batches of Abelmoscus manihot flower with large differences in components were mixed according to a software calculation method. 2000g of the mixed Abelmoscus manihot flower was taken and extracted using percolation with 18 times 70% ethanol. The ethanol was removed from the Abelmoscus manihot flower extract, diluted with water, and extracted using continuous countercurrent extraction with ethyl acetate. The liquid-material ratio of the extract (i.e., extractant:extractant) was 3:1, and the extract loading rate was 120mL / min. The rotation speed was 60 Hz, the extract had 4 stages, the extract solution was reduced pressure at 50°C to recover the solvent, diluted with water, centrifuged, the supernatant was taken and treated with D101 macroporous resin, the macroporous resin treatment process was as follows: the diameter-to-height ratio of the macroporous resin was 1:6, the loading buffer concentration was 0.15 g raw material / mL, the loading buffer volume was 7 BV, loading was performed at a flow rate of 2 BV / h, impurities were removed using 6 BV of pure water and 3 BV of 5% ethanol at a flow rate of 2 BV / h, elution was performed using 4 BV of 60% ethanol at a flow rate of 2 BV / h to obtain the eluate, which was concentrated, reduced pressure and dried at 60°C to obtain the total solids. Rutin 0.22%, hyperoside It contains 20.58% isoquercitrin 17.81%, hibifolin 16.07%, myricetin 5.15%, quercetin-3'-o-glucoside 17.37%, and quercetin 1.77%, with the peak area equivalent of quercetin-3-O-robinobioside being 0.43%. The mass ratios between each component are shown in Table 2-4 below.

[0064] [Table 2-4]

[0065] Example 3: Preparation of flavonoid-active parts of Abelmoscus manihot flowers Example 3.1 100g of Abelmoskasmanihot flowers were ground into a coarse powder and extracted using percolation with 18 times 60% ethanol to obtain an Abelmoskasmanihot flower extract. Ethanol was removed from the Abelmoskasmanihot flower extract, diluted with water, and extracted using continuous countercurrent with ethyl acetate. The liquid-to-material ratio of the extract (i.e., extractant:extractate) was 3:1, the loading rate of the extract was 100mL / min, the rotation speed of the extractor was 40Hz, and the number of extract stages was 5. After recovering the solvent by reducing the pressure of the extract solution at 40°C, it was treated with D101 macroporous resin. The procedure was as follows: the diameter-to-height ratio of the macroporous resin was 1:8, the loading buffer concentration was 0.15 g raw material / mL, the loading buffer volume was 8 BV, loading was performed at a flow rate of 2 BV / h, impurities were removed using 6 BV of pure water and 3 BV of 5% ethanol at a flow rate of 2 BV / h, and elution was obtained using 4 BV of 60% ethanol at a flow rate of 3 BV / h to obtain the eluate. After that, the eluate was reduced in pressure at 60°C to recover the ethanol, and then an Abelmoscus manihot flower flavonoid extract was obtained. The content was measured, and the content of the seven components was adjusted by the accession method according to the content measurement results. The content of each component is shown in the table below. The total solid content of the flavonoid active part of the Abelmoscus manihot flower is 3.18 g. The quercetin-3-O-robinobioside content is 0.20~0.50%.

[0066] [Table 3]

[0067] 3.2 The experiment was repeated with 500g of Abelmoscus manihot flower drug according to the preparation method of Example 3.1, and the content data of the two batches obtained are shown in Table 4 below. The quercetin-3-O-robinobioside content is 0.30-0.80%. A indicates "Each component / Total solids (%)", and B indicates "Each component / Isoquercitrin".

[0068] [Table 4]

[0069] Example 4: Preparation of flavonoid-active parts of Abelmoscus manihot flowers Example 4-1 100g of Abelmoskasmanihot flowers were extracted using percolation with 18 times 70% ethanol to obtain an Abelmoskasmanihot flower extract. Ethanol was removed from the Abelmoskasmanihot flower extract, diluted with water, and subjected to continuous countercurrent extraction with n-butanol. The liquid-to-material ratio of the extract (i.e., extractant:extract) was 3:1, the loading rate of the extract was 120 mL / min, the rotation speed of the extractor was 60 Hz, and the number of extraction stages was 5. After recovering the solvent by reducing the pressure of the extract solution at 50°C, it was treated with D101 macroporous resin. The procedure was as follows: the diameter-to-height ratio of the macroporous resin was 1:6, the loading buffer concentration was 0.15 g raw material / mL, the loading buffer volume was 7 BV, loading was performed at a flow rate of 2 BV / h, impurities were removed using 6 BV of pure water and 3 BV of 5% ethanol at a flow rate of 2 BV / h, and eluate was obtained by eluting with 4 BV of 60% ethanol at a flow rate of 2 BV / h. The eluate was then reduced under reduced pressure at 60°C to recover the ethanol and obtain the Abelmoscus manihot flower extract. The content was measured, and the content of the seven components was adjusted by the accession method according to the content measurement results. The content of each component is shown in the table below. The total solid content of the flavonoid active parts of the Abelmoscus manihot flower was 2.05 g.

[0070] [Table 5]

[0071] Example 4-2 Abelmoscus manihot flowers were extracted using percolation with 18 times 70% ethanol. Ethanol was removed from the extract until the amount was 0.5-1.0 g / mL of raw material. ZTC1+1-II fining agents were added in amounts of 4% component B and 2% component A, respectively. The water bath temperature was 60°C, and the water bath holding time was 60 minutes. The extract was centrifuged, filtered, diluted, and treated with polyamide resin. The resin treatment process was as follows: the diameter-to-height ratio of the resin was 1:7, the loading buffer concentration was 0.15-0.5 g / mL of raw material, and the loading buffer volume was 8 BV. The eluate was obtained by eluting with 5 BV of pure water and 5 BV of 80% ethanol, concentrated, dried under reduced pressure at 60°C, and the content was measured. The content of the seven components was adjusted by the accession method according to the content measurement results. The content of each component is shown in the table below. The total solid content of the flavonoid active parts of Abelmoscus manihot flowers was obtained, and the quercetin-3-O-robinobioside content was 0.39%.

[0072] [Table 5-1]

[0073] Example 5. Preparation of flavonoid-active parts of Abelmoscus manihot flowers Example 5-1 100g of Abelmoskasmanihot flowers were extracted using percolation with 15 times 70% ethanol to obtain an Abelmoskasmanihot flower extract. Ethanol was removed from the Abelmoskasmanihot flower extract, diluted with water, and subjected to continuous countercurrent extraction with n-butanol. The liquid-to-material ratio of the extract (i.e., extractant:extract) was 2.5:1, the loading rate of the extract was 80 mL / min, the rotation speed of the extractor was 50 Hz, and the number of extract stages was 4. After recovering the solvent by reducing the pressure under 50°C, the extract was treated with D101 macroporous resin. The processing steps were as follows: the diameter-to-height ratio of the macroporous resin was 1:6, the loading buffer concentration was 0.2 g raw material / mL, the loading buffer volume was 6 BV, loading was performed at a flow rate of 2 BV / h, impurities were removed using 1 BV of pure water and 4 BV of 5% ethanol at a flow rate of 2 BV / h, elution was obtained using 3 BV of 60% ethanol at a flow rate of 2 BV / h, the eluate was obtained by reducing the pressure of the eluate at 60°C to recover the ethanol, and then the Abelmoscus manihot flower extract was obtained. The content was measured, and the content of the seven components was adjusted by the accession method according to the content measurement results. The content of each component is shown in the table below. The total solid content of the flavonoid active parts of the Abelmoscus manihot flower was 2.86 g.

[0074] Example 5-2 Abelmoscus manihot flowers were reflux-extracted with 15 times 70% ethanol, and ethanol was removed from the extract until the amount of raw material / mL was 0.5-1.0 g. A 10% hydrochloric acid solution was added to adjust the pH to 2.0-3.0. The extract was refrigerated, filtered, washed, and the precipitate was removed. Water was added to dissolve the precipitate, and continuous countercurrent extraction was performed with ethyl acetate. The liquid-to-material ratio of the extract (i.e., extractant:extractate) was 3:1, and the number of extracts was 4. The extract was then reduced under reduced pressure at 50°C to recover the solvent, diluted with water, and treated with polyamide resin. The resin treatment process was as follows: the diameter-to-height ratio of the resin was 1:6, the loading buffer concentration was 0.15-0.5 g raw material / mL, and the loading buffer volume was 6 BV. The eluate was obtained by eluting with 4 BV of pure water and 4 BV of 70% ethanol, concentrated, and dried under reduced pressure at 60°C. The quercetin-3-O-robinobioside content is 0.47%.

[0075] [Table 5-2]

[0076] Example 6. Preparation of flavonoid-active parts of Abelmoscus manihot flowers 100g of Abelmoskasmanihot flowers were extracted by percolation with 16 times 60% ethanol to obtain an Abelmoskasmanihot flower extract. Ethanol was removed from the Abelmoskasmanihot flower extract, diluted with water, and extracted by continuous countercurrent extraction with ethyl acetate. The liquid-to-material ratio of the extract (i.e., extractant:extractate) was 2:1, the loading rate of the extract was 100mL / min, the rotation speed of the extractor was 40Hz, and the number of extract stages was 4. After recovering the solvent by reducing the pressure of the extract solution at 50°C, it was treated with D101 macroporous resin. The treatment process for the macroporous resin was as follows: The diameter-to-height ratio of the macroporous resin was The ratio was 1:6, the loading buffer concentration was 0.3 g raw material / mL, the loading buffer volume was 3 BV, loading was performed at a flow rate of 2 BV / h, impurities were removed using 3 BV of pure water and 3 BV of 5% ethanol at a flow rate of 2 BV / h, and elution was obtained using 4 BV of 70% ethanol at a flow rate of 2 BV / h to obtain the eluate. The content was measured, and the content of seven specific components was adjusted by adding them. The content of each component is shown in the table below. After that, the eluate was reduced under reduced pressure at 60°C to recover the ethanol and obtain the Abelmoscus manihot flower extract. The content was measured, and the content of the seven components was adjusted by the accession method according to the content measurement results. The amount added was 10% or less of the total weight. The content of each component is shown in the table below. The total solid content of the flavonoid active parts of the Abelmoscus manihot flower was 3.16 g.

[0077] [Table 6]

[0078] Example 7. Preparation of flavonoid-active parts from Abelmoscus manihot flowers (Comparative example) Example 7-1 500g of Abelmoskasmanihot flowers were ground into a coarse powder and extracted by percolation with 20 times 70% ethanol to obtain an Abelmoskasmanihot flower extract. After recovering the solvent under reduced pressure, the extract was treated with D101 macroporous resin. The macroporous resin treatment process was as follows: the diameter-to-height ratio of the macroporous resin was 1:4, the loading buffer concentration was 0.15g raw material / mL, the loading buffer volume was 6BV, loading was performed at a flow rate of 2BV / h, impurities were removed using 7BV of pure water and 4B of 5% ethanol at a flow rate of 2BV / h, and elution was obtained using 4BV of 60% ethanol at a flow rate of 1.5BV / h to obtain the eluate. After recovering the ethanol by reducing pressure at 60°C, a total solid content of 16.12g of Abelmoskasmanihot flowers was obtained. The content of each component is shown in the table below.

[0079] [Table 7-1]

[0080] Example 7-2 100g of Abelmoskasmanihot flowers were ground into a coarse powder and extracted using percolation with 18 times 70% ethanol to obtain an Abelmoskasmanihot flower extract. After recovering the solvent under reduced pressure, the extract was treated with D101 macroporous resin. The macroporous resin treatment process was as follows: the diameter-to-height ratio of the macroporous resin was 1:5, the loading buffer concentration was 0.15g raw material / mL, the loading buffer volume was 6BV, loading was performed at a flow rate of 2BV / h, impurities were removed using 6BV of pure water and 3B of 5% ethanol at a flow rate of 2BV / h, and eluate was obtained using 4BV of 60% ethanol at a flow rate of 1.5BV / h to obtain the eluate. After recovering the ethanol by reducing pressure at 60°C, a total solid content of 3.56g of Abelmoskasmanihot flowers was obtained.

[0081] [Table 7-2]

[0082] Example 8. Preparation of flavonoid-active parts from Abelmoscus manihot flowers (comparative example) 100g of Abelmoskasmanihot flowers were ground into a coarse powder and extracted using 18-fold percolation with 70% ethanol to obtain an Abelmoskasmanihot flower extract. Ethanol was removed from the Abelmoskasmanihot flower extract, diluted with water, and extracted continuously in countercurrent with ethyl acetate. The liquid-to-material ratio of the extract (i.e., extractant:extractate) was 2.5:1, the loading rate of the extract was 80 mL / min, the rotation speed of the extractor was 40 Hz, and the number of extraction stages was 4. The extract solution was then cooled under reduced pressure at 40°C to obtain a solvent. After recovery, the material was treated with D101 macroporous resin, and the processing steps for the macroporous resin were as follows: the diameter-to-height ratio of the macroporous resin was 1:5, the loading buffer concentration was 0.15 g raw material / mL, the loading buffer volume was 6 BV, loading was performed at a flow rate of 2 BV / h, impurities were removed using 6 BV of pure water and 3 BV of 5% ethanol at a flow rate of 2 BV / h, and elution was obtained using 4 BV of 60% ethanol at a flow rate of 1.5 BV / h to obtain the eluate, the content was measured, and then rutin was added to the eluate. hyperoside The following was added. The content of each component is shown in the table below. After that, the ethanol was recovered by reducing the pressure at 60°C, and a total solid content of 4.28 g of Abelmoscus manihot flowers was obtained.

[0083] [Table 8]

[0084] Example 9. 9.1. Preparation of flavonoid-active parts of Abelmoscus manihot flowers 25 kg of Abelmoskasmani Hot flowers were ground into a coarse powder and extracted using 18-fold percolation with 70% ethanol to obtain an Abelmoskasmani Hot flower extract. Ethanol was removed from the Abelmoskasmani Hot flower extract, diluted with water, and extracted continuously in countercurrent with ethyl acetate. The liquid-material ratio of the extract (i.e., extractant:extract) was 2.5:1, and the extract had 4 extraction stages. After recovering the solvent by reducing the pressure under 40°C, the extract was treated with D101 macroporous resin. The macroporous resin treatment process was as follows: the diameter of the macroporous resin... The ratio to height was 1:5, the loading buffer concentration was 0.15 g raw material / mL, the loading buffer volume was 3 BV, loading was performed at a flow rate of 1.5 BV / h, impurities were removed at a flow rate of 2 BV / h using 6 BV of pure water and 3 BV of 10% ethanol, and elution was obtained by eluting at a flow rate of 1.5 BV / h using 4 BV of 60% ethanol. The eluate was then reduced under pressure at 60°C to recover the ethanol, and the total solids of Abelmoscus manihot flowers were obtained. Seven specific components were added to prepare the seven components (the content of the added components was 20% or less of the total weight). The content of each component is shown in the table below. The total solids of the flavonoid active parts of Abelmoscus manihot flowers were obtained. The quercetin-3-O-robinobioside content was 0.20-0.50%.

[0085] [Table 9]

[0086] 9.2. Preparation of flavonoid-active parts of Abelmoscus manihot flowers Referring to Example 2-B, 30 kg of Abelmoskasmanihot flower mixture was ground into a coarse powder and extracted by percolation with 18 times 70% ethanol to obtain Abelmoskasmanihot flower extract. Ethanol was removed from the Abelmoskasmanihot flower extract, diluted with water, and extracted by continuous countercurrent extraction with ethyl acetate. The liquid-to-material ratio of the extract (i.e., extractant:extract) was 2.5:1, the loading rate of the extract was 80 mL / min, the rotation speed of the extractor was 40 Hz, and the number of extract stages was 4. After recovering the solvent by reducing the pressure under 40°C, the extract was treated with D101 macroporous resin. The processing steps for the macroporous resin were as follows: the diameter-to-height ratio of the macroporous resin was 1:5, the loading buffer concentration was 0.15 g raw material / mL, the loading buffer volume was 5 BV, loading was performed at a flow rate of 1.5 BV / h, impurities were removed at a flow rate of 2 BV / h using 3 BV of pure water and 4 BV of 5% ethanol, and elution was obtained by eluting at a flow rate of 1.5 BV / h using 5 BV of 60% ethanol. The eluate was then reduced in pressure at 65°C to recover the ethanol, and the total solids of Abelmoscus manihot flowers were obtained. The eluate was vacuum dried, crushed, and the flavonoid content (%) of seven components was detected. The results are shown below, where the quercetin-3-O-robinobioside content was 0.20-0.50%.

[0087] [Table 10]

[0088] Example 11. Preparation of eye drops containing the flavonoid active ingredient of Abelmoscus manihot flower. Preparation process: The active site was prepared according to one of the methods in Examples 1 to 7, dissolved by stirring with sterile water for injection, diluted to 100 L, pH adjusted to 7, filtered, filled, and sterilized to obtain the eye drops. The pH adjusting agent used was sodium hydroxide and / or hydrochloric acid.

[0089] Example 12. Pharmacological experiments on a glucose-releasing kidney model. Example 12-1 Animal model: Streptozotocin (STZ) + high-fat diet glucose kidney model, C57BL / 6 mice, 20±10g, SPF grade, male. Method of administration: Oral administration Experimental Modeling: After acclimatizing experimental mice for 3 days, they were fed a high-fat, high-carbohydrate diet (laboratory-made, recipe: lard:sucrose:egg yolk:basic feed = 18:20:3:59) for 8 weeks. Then, they were intraperitoneally injected with 100 mg / kg of STZ sodium citrate solution for 7 days. On the 8th day, the mice's blood glucose levels were detected via tail vein, and a blood glucose level exceeding 16.7 mmol / L was used as the criterion for creating a diabetes model. A blood glucose level exceeding 13.8 mmol / L, the appearance of proteinuria, and abnormal renal function indicated successful modeling. Experimental grouping: The mice were divided into six groups: a normal group, a model group, a positive drug group (dagliflozin tablets, 45.5 mg / kg), a sample group prepared by the method of Example 2-1 (62.4 mg / kg), a sample group prepared by the method of Example 6 (62.4 mg / kg), and a sample group prepared by the method of Example 8 (62.4 mg / kg). Each group consisted of 15 mice. After 4 weeks of administration, urinary protein and urinary creatinine were detected, and the daily intake and mouse condition were calculated. The detection results are shown in Table 12-1 below.

[0090] [Table 11]

[0091] Each group in the examples had the effect of reducing urinary protein, and the sample group in Example 2-1 was able to significantly reduce both urinary protein and the urinary albumin / urinary creatinine ratio (ACR). As can be seen from the observation results of the mouse condition, the number of mice that developed diarrhea and bloating in the sample group of Example 2 was significantly lower than in the other groups.

[0092] Example 12-2 Animal models: db / db mice, 16 weeks old; C57BL / 6 mice, 16 weeks old. Method of administration: Oral administration Experimental Modeling: After housing db / db mice for 17 weeks, urinary protein was detected. If urinary protein appeared and renal function became abnormal, the model was considered successful and used in subsequent experiments. Experimental grouping: C57BL / 6 mice were divided into a normal group, db / db mice into a model group, a positive drug group (dagliflozin tablets, 45.5 mg / kg), a sample group prepared by the method of Example 3-1 (62.4 mg / kg), a sample group prepared by the method of Example 9.1 (62.4 mg / kg), and a sample group prepared by the method of Example 2-A3 (62.4 mg / kg), for a total of 6 groups, with 15 mice in each group. After 4 weeks, urinary protein and urinary creatinine, as well as daily intake, were detected, and the condition of the mice was observed. The detection results are shown in Table 12-2 below.

[0093] [Table 12]

[0094] Each group in the examples had the effect of reducing urinary protein, and each sample group was able to reduce urinary protein and the urinary albumin / urinary creatinine ratio (ACR). As can be seen from the observation results of the mouse condition, the number of mice that developed diarrhea and bloating in the sample groups of Example 3-1 and Example 9-1 was significantly lower than in the other groups.

[0095] Example 13. Effects of flavonoids on ocular blood flow and choroidal neovascularization. Laboratory animals: Rats, weighing 150-180g, SPF grade, roughly half male and half female. Method of administration: Oral administration Experimental Modeling: Healthy male Brown-Norway rats were selected, anesthetized by intramuscular injection of ketamine (50 mg / kg), and had their pupils dilated with a mixture of 0.5% tropicamide and neoforin. Using a krypton yellow laser (Kojima Corporation, USA) (wavelength 568 nm, parameters: spot diameter 100 μm, exposure time 0.1 s, power 150-200 mW), eight photocoagulation points were performed on the retina between the optic disc, 2-4 disc diameters, peridiscal area, and between major blood vessels, under a 120D pre-lens. A photocoagulation qualifying point was defined as one where Bruch's membrane ruptured and small bubbles occurred; retinal, choroidal, and vitreous hemorrhages were excluded. Experimental grouping: Drug grouping and dosage: Each group consisted of 15 animals, divided into a normal group, a model group, a positive control group (vitexofen 1.35 mg / kg), an Example 2-1 sample group (69.9 mg / kg), an Example 3.1 sample group (87.36 mg / kg), an Example 7 sample group (87.36 mg / kg), an Example 9.1 sample group (87.36 mg / kg), and an Example 9.2 sample group (87.36 mg / kg, solid content prepared according to Example 9.2). The rate of inhibition of choroidal blood flow and choroidal neovascularization area were detected after 4 weeks of administration. The detection results are shown in Table 13 below.

[0096] [Table 13]

[0097] Conclusion: The samples from each example exhibited inhibitory effects on choroidal blood flow inhibition and choroidal neovascularization area, and the sample groups from Example 2, Example 3, Example 9.1, and Example 9.2 were able to significantly suppress choroidal neovascularization.

[0098] Example 14. Effects of flavonoids on contrast-induced kidney damage Experimental animals: SD rats, weighing 200-250g, SPF grade, roughly half male and half female. Method of administration: Oral administration Experimental Modeling: Healthy male SD rats were selected and first administered furosemide (10 ml / kg) intramuscularly. Then, every 15 minutes, a prostaglandin synthesis inhibitor (indomethacin, 10 mg / kg, dissolved in dimethyl sulfoxide and diluted with 1.25% NAHCO3 solution), a nitric oxide synthase inhibitor (N-nitro-L-arginine methyl ester, L-NAME, 10 mg / kg, dissolved in 0.9% NaCl solution), and 76% pantethine (10 ml / kg) were administered via tail vein injection. Modeling Success Criteria The diagnostic criteria for acute kidney injury recommended in the 2012 KDIGOAKI clinical guidelines (diagnosis is made if any of the following conditions are met): (1) serum creatinine elevation exceeding 26.5 μmol / L (0.3 mg / dl) within 48 hours, (2) serum creatinine elevation exceeding 50% of baseline, confirmed or estimated to have occurred within 7 days, and (3) decreased urine output <0.5 ml / (kg*h) lasting for 6 hours or more. Here, in animal models, serum creatinine elevation exceeding 50% of baseline is often used as the diagnostic criterion for acute kidney injury, i.e., the modeling success criterion. Drug grouping and dosage: The following groups consisted of 15 animals each: a normal group, a model group, a positive control group (irbesalta 50 mg / kg / d), a sample group from Example 2-1 (69.9 mg / kg), a sample group from Example 3.1 (87.36 mg / kg), a sample group from Example 7 (87.36 mg / kg), a sample group from Example 9.1 (87.36 mg / kg), and a sample group from Example 9.2 (87.36 mg / kg, solid content prepared in Example 9.2). After 4 weeks of administration, the rate of suppression of choroidal blood flow in the eye and the area of ​​choroidal neovascularization were detected. The detection results are shown in Table 14 below.

[0099] [Table 14]

[0100] Conclusion: Serum creatinine and serum urea nitrogen levels in the model rats with contrast-induced renal damage were significantly higher than in each administration group. In the sample groups of Example 9.1, Example 2-A1, Example 3.1, Example 8, and Example 6, serum creatinine and serum urea nitrogen levels decreased to varying degrees.

[0101] Example 15. Effects of flavonoids on lupus erythematosus-related lupus nephritis. Twenty-six male MRL / lpr mice (lupus erythematosus mice) were selected and observed as one group. They were 13 weeks old, weighed 18-22g, and purchased from Nanjing Junke Biotechnology Co., Ltd. Twenty-four male C57BL / 6 mice were selected as the normal group. They were 13 weeks old, weighed 18-22g, and purchased from Wuhan Hualianke Biotechnology Co., Ltd. Drug grouping and dosage: 15 mice were assigned to each of the following groups: normal group, MRL / lpr mouse model group, positive control group (dexamethasone 1 mg / kg), Example 2-B sample group (69.9 mg / kg), Example 3.1 sample group (87.36 mg / kg), Example 7 sample group (87.36 mg / kg), Example 9.1 sample group (87.36 mg / kg), and Example 8 sample group (87.36 mg / kg, solid content prepared in Example 9.2). Each group was administered for 4 weeks. Experimental indicators: After administration, urine samples were collected 24 hours later, and urinary protein was detected. After collecting the urine, the eyeballs of all mice were removed, blood was collected, and serum was obtained by centrifugation and used for detection of antinuclear antibodies. Simultaneously, the kidneys were removed, rapidly frozen in liquid nitrogen, and then stored in a refrigerator at minus 80 degrees Celsius for use in renal immunoassay detection. The detection results are shown in Table 15 below.

[0102] [Table 15]

[0103] Conclusion: The model group of lupus erythematosus mice showed significantly higher levels of urinary protein, serum antinuclear antibody levels, and renal tissue IgG and C3 deposition compared to each treatment group. Samples from Example 9.1, Example 2-B, Example 3.1, Example 8, and Example 7 all showed improvement in urinary protein, serum antinuclear antibody levels, and renal tissue IgG and C3 deposition, with Samples from Example 2-B, Example 3.1, and Example 5-2 being more effective.

[0104] Example 16. Toxic effects of flavonoids Laboratory animals: Rats, weighing 150-180g, SPF grade, roughly half male and half female. Method of administration: Oral administration Drug grouping and dosage: Normal group, Abelmosque capsule group (8 g / kg extract), Example 2-A1 sample group (5 g / kg), Example 9.1 sample group (8 g / kg). 30 animals in each group were administered once daily for 12 consecutive weeks, with the dosage calculated according to the extract.

[0105] Experimental results: After administration, kidney weight, kidney volume, and organ index were elevated in the Abelmosque capsule group compared to the normal group. Pathological examination revealed moderate renal tubular hypertrophy in the Abelmosque capsule group, but no renal tubular hypertrophy was detected pathologically in the Example 9.2 sample group and Example 2-A1 group. The detection results are shown in Table 16 and Figure 1 below.

[0106] [Table 16]

[0107] Example 17: Action of total flavonoid active sites on idiopathic pulmonary fibrosis Laboratory animals: 60 Kunming mice, half male and half female, weighing 18-22 kg, clean grade. Animal grouping: The mice were randomly divided into six groups: a normal group, a model group, a control group (rosiglitazone 5 mg / kg), a sample group prepared by the method of Example 2-A3 (70 mg / kg), a sample group prepared by the method of Example 9.2 (70 mg / kg), and an Abelmosque capsule sample group (180 mg / kg in extract equivalent). Ten mice were used in each group. Experimental Modeling: After acclimatizing experimental mice for 3 days, they were anesthetized by intraperitoneal injection of 4% chloral hydrate (0.01 ml / g). The mice were fixed in a supine position, routine disinfection was performed, an incision was made in the midline of the neck, and the trachea was exposed by blunt dissection. Bleomycin (5 mg / kg) was slowly injected into the model group, control group, and treatment group via tracheal cartilage ring gap puncture, while the normal control group received the same amount of saline. Immediately after drug injection, the mice were rotated in an upright position for 3-5 minutes to ensure uniform distribution of the drug solution to both lungs, the skin was sutured, the suture site was disinfected, the mice were awakened, and they were housed in a clean-grade observation room. Administration method: Administration was started on day 2 of the modeling study. The treatment group received the above dosage once daily, while the normal group and the model group received the same amount of distilled water (10 ml / kg) in the same manner for 28 consecutive days. Lung histopathological staining: After administration, the right lung was excised, fixed, embedded, and sectioned, and stained with HE according to standard pathological methods. The degree of alveolitis and pulmonary fibrosis was classified into four stages.

[0108] The experimental results are shown in Table 17 below.

[0109] [Table 17]

[0110] As the experimental results show, the degree of alveolitis and pulmonary fibrosis in the lung tissue of the model group of mice significantly increased, exhibiting typical parenchymal lesions of pulmonary fibrosis. After 28 days of administration, the lung index of mice in each dose group significantly decreased, and the degree of alveolitis and pulmonary fibrosis in the lung tissue significantly decreased. The total flavonoid active site significantly reduced the degree of alveolitis and pulmonary fibrosis in mice with pulmonary fibrosis, and has a protective effect on the lungs of mice with pulmonary fibrosis, reducing damage to the lung tissue in fibrotic mice.

[0111] Example 18: Study of the mechanism of action of SGLT2 Sodium-glucose contransporter 2 (SLC5A2, also known as SGLT2) is a gene that encodes a sodium-glucose cotransporter that reabsorbs more than 90% of glucose filtered in the glomeruli. SLC5A2 is expressed only at the brush border within the proximal tubular epithelial cell membrane of the kidney and is considered a marker of the proximal tubule. Its main function is the recycling of filtered sodium and glucose. Inhibiting the expression of the SLC5A2 protein in the renal tissue of patients with diabetes and diabetic nephropathy can protect the renal tissue of patients with diabetes and diabetic nephropathy because it can play a role in the excretion of sodium and glucose. Animal model: db / db mouse, 18 weeks old Method of administration: Oral administration Experimental group division: db / db mice with UACR > 200 mg / g were divided into three groups: a diabetic nephropathy (DKD) group, an Abelmosque capsule group (DKD + HK, 0.84 g / kg / d), and a total flavonoid group (DKD + HT, sample from Example 2-A2, 0.0755 g / kg / d). The Abelmosque capsule group and the total flavonoid group were administered for four weeks (28 days). Microalbumin (UACR) was detected in the urine of db / db mice using the enzyme-linked immunosorbent assay (ELISA), SLC5A2 mRNA expression levels were detected using the reverse transcription polymerase chain reaction (RT-PCR), and SLC5A2 protein expression levels were detected using the immunohistochemistry (IHC) method.

[0112] Experimental results: Compared to the DKD group, there were no significant differences in pre-group body weight for DKD (N=5), HK (N=5), and HT (N=6), or in body weight after 1 week and 4 weeks of administration for DKD (N=5), HK (N=5), and HT (N=6). There were no significant differences in blood glucose after 1 week and 4 weeks of administration for DKD (N=5), HK (N=5), and HT (N=6). There were no significant differences in UACR values ​​after 1 week of administration for DKD (N=5), HK (N=5), and HT (N=6). After 4 weeks of administration, the HT group showed a significant decrease in UACR values ​​compared to the DKD group (*P<0.05), indicating a statistically significant difference. There were no significant differences in renal index for DKD (N=5), HK (N=5), and HT (N=6). Renal index was divided into DKD (N=5), HK (N=5), and HT (N=6). A significant increase in renal index was observed when *P<0.05, indicating a statistically significant difference. As shown in Figure 2, in the DKD group, some glomeruli were lobulated, the basement membrane was significantly thickened, and the glomerular capillaries were significantly compressed. In the mesangial region, proliferation of mesangial cells was observed, and there was a clear expansion of the mesangial stroma. In the HK group, the thickening of the glomerular basement membrane was significantly improved, the glomerular capillaries remained intact, and the proliferation of mesangial cells and mesangial matrix was significantly improved. In the HT group, the glomerular basement membrane was significantly thinned, the glomeruli did not proliferate, the glomerular capillary stroma did not proliferate, and the proliferation of mesangial cells and mesangial matrix did not increase. As shown in Figure 3, quantitative results using IHC-P confirmed that the expression level of SLC5A2 protein was significantly lower in the Abelmosque HK group (N=5) and HT group (N=6) compared to the DKD group (N=5). RT-PCR and immunohistochemistry results also showed that the mRNA expression level of SLC5A2 was significantly lower in the Abelmosque HK group (N=5) and HT group (N=6) compared to the DKD group (N=5). Compared to the DKD group (N=5), the ratio of SLC5A2 IHC-P quantitative expression level to glomerular number was significantly lower in the Abelmosque HK group (N=5) and HT group (N=6), which was consistent with the IHC-P quantitative results.

[0113] Therefore, Abelmosque capsules and total flavonoids can effectively reduce microalbuminuria in db / db mice. Abelmosque capsules and total flavonoids can inhibit the expression of SLC5A2 protein on the cell membrane of proximal tubular cells, thereby reducing the expression of SLC5A2 protein in the kidneys of DKD mice, effectively inhibiting the activity of SLC5A2 protein, and thus effectively reducing glucose reabsorption in the kidneys of DKD mice.

[0114] The above embodiments are merely specific and detailed representations of some embodiments of the present invention and should not be construed as limiting the scope of the patent of the present invention. Those skilled in the art should note that various modifications and improvements are possible without departing from the concept of the present invention, and all such modifications and improvements are included within the scope of protection of the present invention. Therefore, the scope of patent protection of the present invention shall be as defined by the appended claims.

[0115] (Note) (Note 1) It contains flavonoid components in the following mass ratios: The mass ratio of quercetin-8-O-β-D-glucuronide:hypericin:isoquercitrin:myricetin:quercetin-3'-O-glucoside is 10:3.0~20:4.0~18:1.9~6.0:6.0~20. The flavonoid active part of the Abelmoscus manihot flower is characterized by the following features.

[0116] (Note 2) The mass ratio of quercetin-8-O-β-D-glucuronide:hypericin:isoquercitrin:myricetin:quercetin-3'-O-glucoside is 10:4.0~17:4.50~16:1.9~5.5:7.0~16, and preferably 10:6.0~15:5.0~13:2.0~5.0:8.0~14. Furthermore, it contains quercetin, with the ratio of quercetin-8-O-β-D-glucuronide:quercetin being 10:1.0 to 10.0, preferably 10:1.0 to 6.0, and preferably 10:1.5 to 5.0. Furthermore, it contains rutin, with quercetin-8-O-β-D-glucuronide:rutin being 10:0.05~0.6, preferably 10:0.1~0.4, and more preferably 10:0.1~0.3. More preferably, it contains quercetin-3-O-robinobioside, and the ratio of quercetin-8-O-β-D-glucuronide to quercetin-3-O-robinobioside is 10:0.1~2.5, 10:0.1~0.9, and preferably 10:0.15~0.5. The effective site as described in Appendix 1, characterized by the features described herein.

[0117] (Note 3) The following mass ratios [Table 18] A flavonoid active site characterized by containing flavonoid components.

[0118] (Note 4) The following mass ratios [Table 19] Preferably, [Table 20] , moreover, [Table 21] , moreover, [Table 22] , moreover, [Table 23] ,or [Table 24] ,or [Table 25] The effective site as described in Appendix 3, characterized by containing flavonoid components.

[0119] (Note 5) The active site according to any one of appendices 3 to 5, characterized in that the mass ratio value of isoquercitrin is 1.2 or 1.0.

[0120] (Note 6) It further contains rutin, and the mass ratio of isoquercitrin to rutin is 1:0.001~0.08, preferably 1:0.005~0.06, preferably 1:0.006~0.05, preferably 1:0.007~0.04, preferably 1:0.009~0.04, or preferably 1:0.008~0.03, preferably 1:0.009~0.03. Furthermore, it contains quercetin-3-O-robinobioside, and the mass ratio of isoquercitrin to quercetin-3-O-robinobioside is 10:0.1~2.5, 10:0.1~0.9, and preferably 10:0.15~0.5. The effective site described in any one of the appendices 3 to 5, characterized by the above.

[0121] (Note 7) The total content of quercetin, quercetin-3'-O-glucoside, myricetin, quercetin-8-O-β-D-glucuronide, isoquercitrin, and hypericin is 55% or more, preferably 60% or more, preferably 65-85%, and preferably 69-82%. Furthermore, the quercetin-3'-o-glucoside content is 12.1-25%, and also 12.6-23%, 13-20%, 13-20%, or 14-25%. Alternatively, the quercetin content may exceed 0.7%, exceed 1.0%, be between 1.0% and 10%, be between 1.5% and 5%, Furthermore, quercetin-8-O-β-D-glucuronide is present in 8.5-30% and 12-23%. The effective site described in any one of the appendices 1 to 5, characterized by the above.

[0122] (Note 8) The total content of quercetin, quercetin-3'-O-glucoside, myricetin, quercetin-8-O-β-D-glucuronide, isoquercitrin, rutin, and hypericin contained in the aforementioned active site is 55% or more, preferably 60% or more, preferably 65-85% or 66-84%, preferably 69-82%, or 69-90%. Furthermore, the quercetin-3'-o-glucoside content is 12.1-25%, and also 12.6-23% or 13-20%. The effective site described in Appendix 7, characterized by the features described herein.

[0123] (Note 9) A plant extract containing flavonoid components, comprising the following components in the following mass amounts: The composition is 10-25% hypericin, 8-19% isoquercitrin, 5-30% quercetin-8-O-β-D-glucuronide, and 12.1-25% quercetin-3'-o-glucoside. Furthermore, it is 12-23% hypericin, 10-17% isoquercitrin, 8.5-25% quercetin-8-O-β-D-glucuronide, and 12.6-23% quercetin-3'-o-glucoside. Furthermore, it is 13-22% hypericin, 11-17% isoquercitrin, 12-21% quercetin-8-O-β-D-glucuronide, and 13-20% quercetin-3'-o-glucoside. A plant extract characterized by the following features.

[0124] (Note 10) The extract according to Appendix 9, characterized in that it contains 2-10% quercetin, preferably 2.5-9% quercetin, preferably 3-8.5% quercetin, more preferably 1.5-5% quercetin, and further contains 2-11% myricetin, preferably 3-7% myricetin, more preferably 3-5% myricetin, and further contains 0.08-2.5% quercetin-3-O-robinobioside, preferably 0.1-1.5%, more preferably 0.1-0.9%, and more preferably 0.1-0.5%, and further, the plant is Abelmoscus manihot or okra flowers, preferably Abelmoscus manihot flowers.

[0125] (Note 11) A flavonoid component-containing Abelmoscus manihot flower extract, comprising the following components in the following mass content: The composition is 8-26% hypericin, 12.0-19.8% isoquercitrin, 8.5-30% quercetin-8-O-β-D-glucuronide, 3.0-4.9% or 5.1-6.0% myricetin, 14-25% quercetin-3'-o-glucoside, and 1.6-4.9% quercetin. Furthermore, it is 11-22% hypericin, 12.0-17% isoquercitrin, 12-23% quercetin-8-O-β-D-glucuronide, 1.6-4.9% or 5.1-9.0% myricetin, 13-22% quercetin-3'-o-glucoside, and 1.4-8% or 1.4-7.8% quercetin. Abelmoscus mani flower extract characterized by the following features.

[0126] (Note 12) It further contains rutin at a mass content of 0.01-1.0%, and then 0.05-0.8%, 0.09-0.8%, and 0.1-0.6%. Furthermore, it contains 0.08-2.5% of quercetin-3-O-robinobioside, preferably 0.1-1.5%, more preferably 0.1-0.9%, and even more preferably 0.1-0.5%. The abelmoscus mani flower extract as described in Appendix 11, characterized by the features described herein.

[0127] (Note 13) The abelmoscus manihot flower extract according to Appendix 12, characterized in that the mass content of flavonoid components in the abelmoscus manihot flower extract is 55% or more, preferably 60% or more, preferably 65-85% or 66-84%, and preferably 69-82% or 69-90%.

[0128] (Note 14) It contains flavonoid components in the following mass ratios: The mass ratios of isoquercitrin:quercetin:quercetin-3'-O-glucoside:myricetin:quercetin-8-O-β-D-glucuronide:hypericin are 1:0.05~1.6:0.2~4.6:0.05~1.2:0.2~3.5:0.25~3.6, and further 1:0.1~1.2:0.4~3.1:0.10~0.9:0.4~3.1:0.5 ~3.0, further 1:0.16~1.0:0.6~2.4:0.14~0.75:0.6~2.5:0.6~2.4, further 1:0.2~0.8:0.8~2.1:0.18~0.6:0.8~2.0:0.7~2.0, further 1:0.2~0.7:0.8~1.6:0.2~0.5:0.8~1.8:0.8~1.4, further 1:0.2~0.6:1 0.0~1.2:0.2~0.4:0.9~1.7:1.0~1.3 or 1.2:0.05~1.6:0.2~4.6:0.05~1.2:0.2~3.5:0.25~3.6, and further 1:0.1~1.2:0.4~3.1:0.10~0.9:0.4~3.1:0.5~3.0, and further 1:0.16~1.0:0.6~2.4:0.14~0.75: A composition with the following ratios: 0.6~2.5:0.6~2.4, further 1:0.2~0.8:0.8~2.1:0.18~0.6:0.8~2.0:0.7~2.0, further 1:0.2~0.7:0.8~1.6:0.2~0.5:0.8~1.8:0.8~1.4, and further 1:0.2~0.6:1.0~1.2:0.2~0.4:0.9~1.7:1.0~1.3.

[0129] (Note 15) The composition further contains rutin, and the mass ratio of isoquercitrin to rutin is 1:0.001 to 0.08, preferably 1:0.005 to 0.06, preferably 1:0.006 to 0.05, preferably 1:0.007 to 0.04, preferably 1:0.008 to 0.03, preferably 1:0.009 to 0.03. Furthermore, the quercetin-3'-o-glucoside content in the flavonoid components is 12.1-25%, and even further, the quercetin-3'-o-glucoside content is 12.6-23% or 13-20%. Furthermore, it contains 0.08-2.5% of quercetin-3-O-robinobioside, preferably 0.1-1.5%, more preferably 0.1-0.9%, and even more preferably 0.1-0.5%. The composition described in Appendix 14, characterized by the features described herein.

[0130] (Note 16) The composition according to Appendix 15, characterized in that the total content of quercetin, quercetin-3'-O-glucoside, myricetin, quercetin-8-O-β-D-glucuronide, isoquercitrin, rutin, and hypericin contained in the composition is 55% or more, preferably 60% or more, preferably 65-85%, and preferably 69-82%.

[0131] (Note 17) A pharmaceutical composition comprising an active site described in any one of Appendix 1 to 8, an Abelmoscus manihot extract described in any one of Appendix 9 to 13, or a composition described in any one of Appendix 14 to 16, and further comprising a pharmaceutically acceptable carrier.

[0132] (Note 18) The pharmaceutical composition according to Appendix 17, characterized in that the pharmaceutically acceptable carrier comprises a solvent, an emulsifier, a disintegrant, a filler, a solubilizer, an antioxidant, a pH adjuster, an osmotic pressure adjuster, a bacteriostatic agent, a diluent, a lubricant, a binder and / or a film-forming agent, wherein the lubricant does not contain magnesium stearate.

[0133] (Note 19) The pharmaceutical composition according to Appendix 18, characterized in that the dosage form of the pharmaceutical composition is an injection, a tablet, a suppository, an ointment, a gel, a pill, a tablet, a granule, a capsule, or a compound, a suspension, or a powder.

[0134] (Note 20) Uses of the active site described in any one of Appendix 1 to 8, the Abelmoscus manihot extract described in any one of Appendix 9 to 13, the composition described in any one of Appendix 14 to 16, or the pharmaceutical composition described in any one of Appendix 17 to 19 in the preparation of drugs for nephropathy, preferably diabetic nephropathy or diabetic nephropathy or nephritis accompanied by renal fibrosis.

[0135] (Note 21) Uses of an active site described in any one of Appendix 1 to 8, an Abelmoscus manihot extract described in any one of Appendix 9 to 13, a composition described in any one of Appendix 14 to 16, or a combination of drugs described in any one of Appendix 17 to 19 in the preparation of ophthalmic disease drugs, preferably for macular degeneration, visual fatigue or cataracts, diabetic retinopathy, age-related macular degeneration, central serous exudative chorioretinitis, and macular degeneration due to high myopia.

[0136] (Note 22) Uses of the active site described in any one of Appendix 1 to 8, the Abelmoscus manihot extract described in any one of Appendix 9 to 13, the composition described in any one of Appendix 14 to 16, or the combination of drugs described in any one of Appendix 17 to 19 in the preparation of drugs for lupus erythematosus nephritis, contrast-induced renal impairment, pulmonary fibrosis, heart failure, or uric acid reduction.

[0137] (Note 23) A method for preparing an Abelmoscus manihot extract described in any one of Appendix 1 to 8 or any one of Appendix 9 to 13, (1) A step of preparing an extract by extracting the Abelmoscus manihot flower or the medicinal part of Abelmoscus manihot with ethanol, (2) After concentrating the extract Extraction The step of preparing the extract solution, (3) After removing the solvent from the extract solution, the flavonoid active part or extract of Abelmoscus manihot flower is eluted with a macroporous resin. A preparation method comprising, preferably, percolation as the extraction method.

[0138] (Note 24) The amount of ethanol used in step (1) is 10 to 25 times the amount of the Abelmoscus manihot flower or the medicinal part of the Abelmoscus manihot, and the ethanol is a 60 to 95% ethanol solution. The extractant for the extract in step (2) is n-butanol, petroleum ether, or ethyl acetate, the extraction method is continuous countercurrent extraction, the liquid-material ratio of the extract is 0.8 to 4:1, and the number of extraction stages is 1 to 5. The type of macroporous resin in step (3) is D101, HPD100, or AB-8. The preparation method described in Appendix 23, characterized by the features described herein.

[0139] (Note 25) The preparation method according to Appendix 24, characterized in that step (2) further comprises performing activated carbon adsorption, alcohol precipitation, or acid precipitation treatment on the extract before the extraction.

[0140] (Note 26) The preparation method described in Appendix 25, characterized in that the elution step for the macroporous resin is as follows: the diameter-to-height ratio of the macroporous resin is 1:4 to 1:9, the loading buffer concentration is 0.10 to 0.30 g raw material / mL, the loading buffer volume is 4 to 12 BV, loading is performed at a flow rate of 1 to 4 BV / h, impurities are removed at a flow rate of 0.5 to 4 BV / h using 4 to 8 BV of pure water and 1 to 5 BV of 3 to 15% ethanol, and elution is performed at a flow rate of 1 to 5 BV / h using 2 to 8 BV of 50 to 80% ethanol.

[0141] (Note 27) A method for preparing an Abelmoscus manihot extract described in any one of Appendix 1 to 8 or any one of Appendix 9 to 13, (1) A step of preparing an extract by extracting the Abelmoscus manihot flower or the medicinal part of Abelmoscus manihot with ethanol, (2) Adding a clarifying agent to the extract, treating it in a water bath, and filtering to remove the supernatant, (3) The supernatant is eluted with a polyamide resin to prepare the flavonoid active site or extract of Abelmoscus manihot flower. A preparation method comprising, preferably, percolation as the extraction method.

[0142] (Note 28) The amount of ethanol used in step (1) is 10 to 25 times the amount of the Abelmoscus manihot flower or the medicinal part of the Abelmoscus manihot, and the ethanol is a 60 to 95% ethanol solution. In step (2), the water bath temperature is 50-70°C, and the bathing time is 30-90 minutes. In step (3), the diameter-to-height ratio of the resin is 1:4 to 1:9, the loading buffer concentration is 0.10 to 0.60 g raw material / mL, the loading buffer volume is 4 to 12 BV, and elution is performed using 4 to 8 BV of pure water and 4 to 8 BV of 60 to 95% ethanol. The preparation method described in Appendix 27, characterized by the features described herein.

[0143] (Note 29) A method for preparing an Abelmoscus manihot extract described in any one of Appendix 1 to 8 or any one of Appendix 9 to 13, (1) A step of preparing an extract by extracting the Abelmoscus manihot flower or the medicinal part of Abelmoscus manihot with ethanol, (2) Adjust the pH of the extract to 2.0 to 3.0, refrigerate it, filter it to remove the precipitate, and add water to dissolve it. (3) Dissolve Extraction The step of preparing the extract solution, (4) After removing the solvent from the extract solution, the flavonoid active part or extract of the Abelmoscus manihot flower is eluted with a polyamide resin. A preparation method comprising, preferably, reflux extraction.

[0144] (Note 30) The amount of ethanol used in step (1) is 10 to 25 times the amount of Abelmoscus manihot flower or the medicinal part of Abelmoscus manihot, and the ethanol is a 60 to 95% ethanol solution. The pH adjuster in step (2) is hydrochloric acid, sulfuric acid, acetic acid, phosphoric acid, citric acid, tartaric acid, or maleic acid. The extractant for the extract in step (3) is n-butanol, petroleum ether, or ethyl acetate, the extraction method is continuous countercurrent extraction, the liquid-material ratio of the extract is 0.8 to 4:1, and the number of extraction stages is 1 to 5. In step (4), the diameter-to-height ratio of the resin is 1:4 to 1:9, the loading buffer concentration is 0.10 to 0.60 g raw material / mL, the loading buffer volume is 4 to 12 BV, and elution is performed using 4 to 8 BV of pure water and 4 to 8 BV of 60 to 95% ethanol. The preparation method described in Appendix 29, characterized by the features described herein.

Claims

1. An extract of Abelmoscus manihot flower containing a flavonoid active ingredient, or a composition prepared from the extract of Abelmoscus manihot flower containing the flavonoid active ingredient, wherein the flavonoid active part of Abelmoscus manihot flower is: It contains flavonoid components in the following mass ratios: The mass ratio of goshipetin-8-O-β-D-glucuronide:hyperoside:isoquercitrin:myricetin:quercetin-3'-O-glucoside:quercetin:quercetin-3-O-robinobioside is 10:3.0 to 20:4.0 to 18:1.9 to 6.0:6.0 to 20:1.0 to 10.0:0.1 to 0.9, the quercetin-3'-O-glucoside content is 12.1 to 25%, the goshipetin-8-O-β-D-glucuronide content is 8.5 to 30%, and the quercetin-3-O-robinobioside content is 0.9% or less. The flavonoid active part of the Abelmoscus manihot flower is characterized by the following features.

2. The mass ratio of goshipetin-8-O-β-D-glucuronide:hyperoside:isoquercitrin:myricetin:quercetin-3'-O-glucoside is 10:4.0 to 17:4.50 to 16:1.9 to 5.5:7.0 to 16, the ratio of goshipetin-8-O-β-D-glucuronide:quercetin is 10:1.0 to 6.0, it contains rutin, the ratio of goshipetin-8-O-β-D-glucuronide:rutin is 10:0.05 to 0.6, and the ratio of goshipetin-8-O-β-D-glucuronide:quercetin-3-O-robinobioside is 10:0.15 to 0.

5. The flavonoid active site according to feature 1.

3. The mass ratio of goshipetin-8-O-β-D-glucuronide:hyperoside:isoquercitrin:myricetin:quercetin-3'-O-glucoside:quercetin:rutin:quercetin-3-O-robinobioside is 10:6.0 to 15:5.0 to 13:2.0 to 5.0:8.0 to 14:1.5 to 5.0:0.1 to 0.3:0.1 to 0.9, The total content of quercetin, quercetin-3'-O-glucoside, myricetin, goshipetin-8-O-β-D-glucuronide, isoquercitrin, and hyperoside is 69-82%. The quercetin-3'-O-glucoside content is 13-20%, or 14-25%. The quercetin content is 1.5% to 5%. Goshipetin-8-O-β-D-glucuronide is present in a concentration of 8.5-30%. The quercetin-3-O-robinobioside content is 0.9%. The flavonoid active site according to feature 1.

4. An extract of Abelmoscus manihot flower containing a flavonoid active ingredient, or a composition prepared from the extract of Abelmoscus manihot flower containing the flavonoid active ingredient, wherein the flavonoid active part of Abelmoscus manihot flower is: It contains flavonoid components in the following mass ratios: Table 1 The quercetin-3'-O-glucoside content is 12.1-25%, the quercetin content is higher than 1.0%, and the goshipetin-8-O-β-D-glucuronide content is 8.5-30%. It contains quercetin-3-O-robinobioside, and the mass ratio of isoquercitrin to quercetin-3-O-robinobioside is 10:0.1 to 0.

9. A flavonoid active site characterized by the above.

5. A flavonoid component is contained in the following mass ratio: Table 2 It contains rutin, and the mass ratio of isoquercetin to rutin is 1:0.009 to 0.

04. It contains quercetin-3-O-robinobioside, and the mass ratio of isoquercitrin to quercetin-3-O-robinobioside is 10:0.1 to 0.

9. It contains quercetin, with the total content of quercetin-3'-O-glucoside, myricetin, goshipetin-8-O-β-D-glucuronide, isoquercitrin, and hyperoside being 69-82%. The quercetin-3'-O-glucoside content is 13-20%, or 14-25%. The quercetin content is 1.5% to 5%. Goshipetin-8-O-β-D-glucuronide is present in a concentration of 8.5-30%. The flavonoid active site as described in feature 4.

6. A flavonoid component is contained in the following mass ratio: Table 3 ,or, Table 4 ,or, Table 5 That is, The flavonoid active site as described in feature 4.

7. The mass ratio of isoquercitrin in the flavonoid active ingredient is 1.2 or 1.0, or It further contains rutin, with a mass ratio of isoquercitrin to rutin of 1:0.001 to 0.08, and a mass ratio of isoquercitrin to quercetin-3-O-robinobioside of 10:0.15 to 0.

5. The flavonoid active site as described in feature 4.

8. The total content of quercetin, quercetin-3'-O-glucoside, myricetin, goshipetin-8-O-β-D-glucuronide, isoquercitrin, and hyperoside is 55% or more, and the quercetin-3'-O-glucoside content is 12.6-23%, 13-20%, or 14-25%. The quercetin content is 1.0% to 10%. Goshipetin-8-O-β-D-glucuronide is present in 12% to 23% of the total. The flavonoid active site according to any one of claims 1 to 7.

9. A plant extract containing flavonoid components, comprising the following components in the following mass amounts: The composition is 12-23% hyperoside, 10-17% isoquercitrin, 8.5-25% goshipetin-8-O-β-D-glucuronide, 12.6-23% quercetin-3'-o-glucoside, 12-21% quercetin, and 0.1-0.9% quercetin-3-O-robinobioside. A plant extract characterized by the following features.

10. A component containing the following mass content: The composition is 13-22% hyperoside, 11-17% isoquercitrin, 12-21% goshipetin-8-O-β-D-glucuronide, and 13-20% quercetin-3'-o-glucoside. Furthermore, the plant in question is either Abelmoscus manihot or the flower of okra. The plant extract according to feature 9.

11. The composition is 13-22% hyperoside, 11-17% isoquercitrin, 12-21% gosypetin-8-O-β-D-glucuronide, 13-20% quercetin-3'-o-glucoside, 1.5-5% quercetin, 3-5% myricetin, and 0.1-0.9% quercetin-3-O-robinobioside, and the plant is Abelmoscus manihot or okra flower. The plant extract according to feature 9.

12. A flavonoid component-containing Abelmoscus manihot flower extract, comprising the following components in the following mass content: The composition is 8-26% hyperoside, 12.0-19.8% isoquercitrin, 8.5-30% goshipetin-8-O-β-D-glucuronide, 3.0-4.9% or 5.1-6.0% myricetin, 14-25% quercetin-3'-o-glucoside, 1.6-4.9% quercetin, and 0.1-0.9% quercetin-3-O-robinobioside. Abelmoscus mani flower extract characterized by the following features.

13. A flavonoid component-containing Abelmoscus manihot flower extract, comprising the following components in the following mass content: The composition is 11-22% hyperoside, 12.0-17% isoquercitrin, 12-23% goshipetin-8-O-β-D-glucuronide, 1.6-4.9% or 5.1-9.0% myricetin, 13-22% quercetin-3'-o-glucoside, 1.4-8% quercetin, and 0.1-0.9% quercetin-3-O-robinobioside. Abelmoscus mani flower extract characterized by the following features.

14. Further containing rutin in a mass content of 0.01 to 1.0%, Furthermore, it contains 0.08-2.5% quercetin-3-O-robinobioside. The mass content of flavonoid components in the Abelmoscus manihot flower extract is 55% or more. The abelmoscus manihot flower extract according to claim 12 or 13, characterized in that it is a feature of the present invention.

15. Further containing the following components in the following mass content: The composition is 11-22% hyperoside, 12.0-17% isoquercitrin, 12-23% goshipetin-8-O-β-D-glucuronide, 1.6-4.9% or 5.1-9.0% myricetin, 13-22% quercetin-3'-o-glucoside, 1.4-8% quercetin, 0.1-0.6% rutin, and 0.1-0.9% quercetin-3-O-robinobioside. The Abelmoscus manihot flower extract according to feature 13.

16. A composition containing flavonoid components in the following mass ratios: The mass ratio of isoquercitrin:quercetin:quercetin-3'-O-glucoside:myricetin:goshipetin-8-O-β-D-glucuronide:hyperoside is 1:0.1-1.2:0.4-3.1:0.10-0.9:0.4-3.1:0.5-3.

0. The total content of quercetin, quercetin-3'-O-glucoside, myricetin, goshipetin-8-O-β-D-glucuronide, isoquercitrin, rutin, and hyperoside contained in the above composition is 69-82%. A composition characterized by the following features.

17. The mass ratio of isoquercitrin:quercetin:quercetin-3'-O-glucoside:myricetin:goshipetin-8-O-β-D-glucuronide:hyperoside is 1:0.16-1.0:0.6-2.4:0.14-0.75:0.6-2.5:0.6-2.4, The composition further contains rutin, and the mass ratio of isoquercitrin to rutin is 1:0.001 to 0.

08. The quercetin-3'-o-glucoside content in the flavonoid components is 12.1-25%. Contains 0.08-2.5% quercetin-3-O-robinobioside. The composition according to claim 16.

18. The mass ratio of isoquercitrin:quercetin:quercetin-3'-O-glucoside:myricetin:goshipetin-8-O-β-D-glucuronide:hyperoside:rutin is 1:0.2-0.6:1.0-1.2:0.2-0.4:0.9-1.7:1.0-1.3:1:0.009-0.03, The quercetin-3'-O-glucoside content is 12.6-23% or 13-20%, and the quercetin-3-O-robinobioside content is 0.1-0.5%. The composition according to claim 16.

19. A pharmaceutical composition comprising a flavonoid active site according to any one of claims 1 to 7, a plant extract according to claims 9 to 11, an Abelmoscus manihot flower extract according to claims 12 and 14, or a composition according to claims 16 to 17, Further comprising a pharmaceutically acceptable carrier, The pharmaceutically acceptable carrier comprises a solvent, emulsifier, disintegrant, filler, solubilizer, antioxidant, pH adjuster, osmotic pressure adjuster, bacteriostatic agent, diluent, lubricant, binder and / or film-forming agent, wherein the lubricant does not contain magnesium stearate. The dosage form of the aforementioned pharmaceutical composition is an injection, tablet, suppository, ointment, gel, pill, tablet, granule, capsule, or compound, suspension, or powder. A pharmaceutical composition characterized by the following features.

20. Use of the pharmaceutical composition according to claim 19, one or more of the following: (1) Use in the preparation of drugs for nephropathy or nephritis, (2) Use in the preparation of ophthalmic drugs, visual fatigue or cataracts, diabetic retinopathy, age-related macular degeneration, central serous chorioretinitis, and macular degeneration due to high myopia. (3) Use in the preparation of drugs for lupus erythematosus nephritis, contrast-induced renal impairment, pulmonary fibrosis, heart failure, or uric acid reduction.

21. Any one or more uses of the flavonoid active part according to any one of claims 1 to 7, the plant extract according to claims 9 to 11, the Abelmoscus manihot flower extract according to claims 12 and 14, or the composition according to claim 16 to 17, as follows: (1) Use in the preparation of drugs for nephropathy or nephritis, (2) Use in the preparation of ophthalmic drugs, visual fatigue or cataracts, diabetic retinopathy, age-related macular degeneration, central serous chorioretinitis, and macular degeneration due to high myopia. (3) Use in the preparation of drugs for lupus erythematosus nephritis, contrast-induced renal impairment, pulmonary fibrosis, heart failure, or uric acid reduction.

22. A method for preparing a flavonoid active site according to any one of claims 1 to 7, a plant extract according to claims 9 to 11, and an Abelmoscus manihot flower extract according to claims 12 and 14, the method being selected from any of the following preparation methods 1 to 3. Preparation method 1 consists of the following steps (1) A step of preparing an extract by extracting the Abelmoscus manihot flower or the medicinal part of Abelmoscus manihot with ethanol, (2) After concentrating the extract, the step of preparing the extract by extraction, (3) After removing the solvent from the extract solution, the flavonoid active part or extract of Abelmoscus manihot flower is eluted with a macroporous resin. A preparation method comprising, The amount of ethanol used in step (1) is 10 to 25 times the amount of the abelmoscus manihot flower or the medicinal part of the abelmoscus manihot, and the ethanol is a 60 to 95% ethanol solution. The extractant for the extraction in step (2) is n-butanol, petroleum ether, or ethyl acetate, the extraction method is continuous countercurrent extraction, the liquid-to-material ratio of the extraction is 0.8 to 4:1, and the number of extraction stages is 1 to 5. The type of the macroporous resin in step (3) is D101, HPD100, or AB-8, and / or Step (2) further comprises, prior to the extraction, performing activated carbon adsorption, alcohol precipitation, or acid precipitation treatment on the extract, and / or The elution process for the macroporous resin is as follows: the diameter-to-height ratio of the macroporous resin is 1:4 to 1:9, the loading buffer concentration is 0.10 to 0.30 g raw material / mL, the loading buffer volume is 4 to 12 BV, loading is performed at a flow rate of 1 to 4 BV / h, impurities are removed using 4 to 8 BV of pure water and 1 to 5 BV of 3 to 15% ethanol at a flow rate of 0.5 to 4 BV / h, and elution is performed using 2 to 8 BV of 50 to 80% ethanol at a flow rate of 1 to 5 BV / h. Preparation method, Preparation method 2 involves the following steps: (1) A step of preparing an extract by extracting the Abelmoscus manihot flower or the medicinal part of Abelmoscus manihot with ethanol, (2) Adding a clarifying agent to the extract, treating it in a water bath, filtering it and removing the supernatant, (3) Eluting the supernatant with a polyamide resin to prepare the flavonoid active part or extract of the Abelmoscus manihot flower, A preparation method comprising, The amount of ethanol used in step (1) is 10 to 25 times the amount of the abelmoscus manihot flower or the medicinal part of the abelmoscus manihot, and the ethanol is a 60 to 95% ethanol solution. In step (2), the water bath temperature is 50-70°C, and the bathing time is 30-90 minutes. In step (3), the diameter-to-height ratio of the resin is 1:4 to 1:9, the loading buffer concentration is 0.10 to 0.60 g raw material / mL, the loading buffer volume is 4 to 12 BV, and elution is performed using 4 to 8 BV of pure water and 4 to 8 BV of 60 to 95% ethanol. Preparation method, Preparation method 3 consists of the following steps (a) A step of preparing an extract by extracting the Abelmoscus manihot flower or the medicinal part of Abelmoscus manihot with ethanol, (b) Adjust the pH of the extract to 2.0 to 3.0, refrigerate it, filter it to remove the precipitate, and add water to dissolve it. (c) A step of extracting the solution to prepare the extract solution, (d) After removing the solvent from the extract solution, the extract is eluted with a polyamide resin to prepare the flavonoid active part or extract of the Abelmoscus manihot flower. A preparation method comprising, The amount of ethanol used in step (a) is 10 to 25 times the amount of Abelmoskasmanihot flower or the medicinal part of Abelmoskasmanihot, and the ethanol is a 60 to 95% ethanol solution. The pH adjuster in step (b) is hydrochloric acid, sulfuric acid, acetic acid, phosphoric acid, citric acid, tartaric acid, or maleic acid. The extractant for the extraction in step (c) is n-butanol, petroleum ether, or ethyl acetate, the extraction method is continuous countercurrent extraction, the liquid-to-material ratio of the extraction is 0.8 to 4:1, and the number of extraction stages is 1 to 5. In step (d), the diameter-to-height ratio of the resin is 1:4 to 1:9, the loading buffer concentration is 0.10 to 0.60 g raw material / mL, the loading buffer volume is 4 to 12 BV, and elution is performed using 4 to 8 BV of pure water and 4 to 8 BV of 60 to 95% ethanol. Preparation method.

23. The total content of quercetin, quercetin-3'-O-glucoside, myricetin, goshipetin-8-O-β-D-glucuronide, isoquercitrin, and hyperoside contained in the flavonoid active site is 55% or more. Furthermore, the quercetin-3'-o-glucoside content is 12.1-25%, Alternatively, if the quercetin content exceeds 0.7%, Furthermore, goshipetin-8-O-β-D-glucuronide is present in 8.5-30% of the total. The preparation method according to feature 22.

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