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]

[0001] The present invention relates to the pharmaceutical field of traditional Chinese medicine, specifically to the flavonoid active site of Abelmoschus maniihot flower and its uses. [Background technology]

[0002] Medicinal Abelmoschus manihot flower is the dried flower crown of the mallow plant Abelmoschus manihot (L.) Medicinal, which has a sweet and cool taste, belongs to the kidney and bladder meridians, has the effects of removing dampness and heat, soothing swelling, and removing toxins, and is used for carbuncles, gangrene, and burns caused by water or fire. This medicine was first listed in the Jiayou Bencao, and was listed in the Jiayou Bencao as follows: Abelmoschus manihot flower can be applied externally to treat those with malignant sores and pus that have not healed for a long time. It was listed in the Compendium of Materia Medica as follows: Abelmoschus manihot flower, its flowers are sweet-smelling, cold, slippery, non-toxic, can be applied externally to treat those with malignant sores and pus that have not healed for a long time, and is the main remedy for patients with chronic sores.

[0003] Flavonoid compounds are not only one of the main components of Abelmoschus manii flower, but also one of its pharmacologically active ingredients. Abelmoschus capsules have been on the market for many years, and its main active ingredient is Abelmoschus manii flower extract. Research has confirmed that the flavonoid components in Abelmoschus manii flower extract have significant therapeutic effects on sores and pus, and also have significant protective effects on the damage to the heart, brain, tissues, etc. caused by ischemia. In addition, it has been reported in many literatures that the total flavonoids in Abelmoschus manii flower may have better effects in the fields of anti-inflammation, antipyretic analgesia, protection against cardiac and cerebral ischemia damage, hypoglycemic, anti-virus, etc. For example, Chinese Patent Application No. 201210082553.7 discloses a total flavonoid extract of Abelmoschus maniifolia flower, which contains quercetin-3'-glucoside, quercetin-3'-glucoside and isoquercitrin with a weight ratio of (11-16):(2.5-6):(4-6.5), and is used for preparing a medicine for treating nephropathy diseases. Chinese Patent Application No. 200610097615.6 discloses a total flavonoid extract from Abelmoschus manii flower, which has a total flavonoid content of 50-90% by weight, and includes 1.0-5.0% quercetin-3-robinobioside, 8-24.0% hypericin, 7.0-20.0% isoquercitrin, 5.0-15.0% quercetin-3'-glucoside, 3.0-10.0% quercetin-3'-glucoside, 0.5-5.0% myricetin, 0.5-5.0% quercetin, 2.0-8.0% quercetin, and several other flavonoid components. The total flavonoid extract from Abelmoschus manii flower is used to prepare a medicine for treating nephropathy, and the quality and quantity of the flavonoid components are clear and the treatment effect is reliable.

[0004] Despite the various therapeutic effects of bioflavonoids, there are numerous types of flavonoids, which can currently be classified into seven different subtypes, and different compositions of flavonoid components have different therapeutic effects. For example, it has been reported in the literature that bioflavonoids are effective in preventing and treating diabetic retinopathy, but compounds with five hydroxyl groups, such as quercetin, have a negative effect on ocular blood flow, whereas dehydrogenating flavonoids to flavanones results in significant improvement of ocular blood flow. Furthermore, compounds that increase blood flow significantly increase retinal function recovery after ischemic damage. How to develop and effectively utilize such compounds to treat ocular diseases such as macular degeneration, visual fatigue, and cataracts, or to enhance their effectiveness against diabetic retinopathy, is an important research issue.

[0005] Choroidal neovascularization (CNV) is seen in many ocular diseases such as age-related macular degeneration (AMD), central exudative chorioretinitis, and macular degeneration due to high myopia, among which CNV secondary to AMD is the most common and is one of the main causes of irreversible visual dysfunction in elderly people. Blood flow insufficiency is also a factor that affects visual fatigue, and promoting ocular blood flow and acting on ocular smooth muscle contributes to the recovery of visual fatigue.

[0006] Proximal renal tubule glucose reabsorption is mediated by sodium-glucose cotransporter protein (SGLT) 1 and SGLT2 transporters, where approximately 90% of glucose reabsorption is mediated by SGLT2 and the remaining 10% is mediated by SGLT1. SGLT2 inhibitors selectively bind to the SGLT2 receptor, thereby inhibiting renal tubule glucose reabsorption, lowering the renal glucose threshold, increasing urinary glucose excretion, and achieving significant blood glucose lowering purposes. In recent years, clinical studies have found that SGLT2 inhibitors have renal protective effects on non-diabetic chronic kidney disease (CKD). SGLT2 inhibitors can reduce the expression of collagen, fibronectin, and reduce the accumulation of extracellular matrix by inhibiting the AGEs-RAGE axis, and alleviate the progression of DN renal fibrosis. SGLT2 inhibitors can regulate the feedback of renal tubules, reduce the process of proteinuria, and in addition to their anti-inflammatory and anti-fibrotic effects, they can also reduce endogenous uric acid (SUA) in diabetic patients. For example, compared with the control group, SGLT2 inhibitors (empagliflozin, cargliflozin, dagliflozin, togliflozin, lugliflozin, and igliflozin) significantly reduced serum uric acid levels. SGLT2 inhibitors may be beneficial in the prevention and treatment of heart failure by reducing plasma volume, reducing pre- and post-load, improving myocardial energy metabolism, and improving myocardial remodeling.

[0007] The methods of extracting Abelmoschus manii flower reported in literature include ethanol reflux extraction, ultrasonic extraction, room temperature soaking extraction, etc., but the components of Abelmoschus manii flower extract are complex, the content of active ingredients varies widely, the extraction transfer rate is low, the energy consumption of extraction is high, and the solvent consumption is high.Therefore, it is necessary to explore Abelmoschus manii flower therapeutic drug and its extraction method that can be combined to simultaneously treat various diseases such as nephropathy, especially diabetic nephropathy and ophthalmology disease.

[0008] The applicant has conducted a statistical study of side effects of Abelmoscu Capsules, and found that there were 194 cases of side effects in the past year. According to the statistics of side effects, nausea, itching, rash, vomiting, diarrhea, epigastric fullness, discomfort, etc. are all known side effects with more than 10 occurrences. Recently emerging ADR symptoms include dizziness, headache, and loss of appetite. In addition, one serious side effect reported was hepatocellular damage, which may suggest hepatocellular damage caused by Abelmoscu Capsules. Therefore, there is clinical significance in developing a new flavonoid extract of Abelmoscu Manihot flower to reduce side effects and prepare drugs for disease treatment. Summary of the Invention [Problem to be solved by the invention]

[0009] In view of the above-mentioned shortcomings, the present invention provides the flavonoid effective part of Abelmoschus manii flower and its use. The present invention uses extract-binding resin method to prepare high-purity Abelmoschus manii flower flavonoid effective part, which has good therapeutic activity in nephropathy, such as diabetic nephropathy, contrast agent-induced renal damage or lupus erythematosus nephritis, and also has the effect of treating ophthalmological diseases. At the same time, the flavonoid effective part or extract or composition of Abelmoschus manii flower described in the present invention has less side effects and can provide patients with better therapeutic drugs. [Means for solving the problem]

[0010] In order to solve the above technical problems, the present invention provides the following technical solutions:

[0011] In a first aspect, the present invention provides a flavonoid active site of Abelmoschus maniihot flower containing flavonoid components in the following mass ratio, the mass ratio of quercetin-8-O-β-D-glucuronide:hypericin:isoquercitrin:myricetin:quercetin-3'-O-glucoside being 10:3.0-20:4.0-18:1.9-6.0:6.0-20.

[0012] Specifically, 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, preferably 10:6.0-15:5.0-13:2.0-5.0:8.0-14, and further contains quercetin, where the ratio of quercetin-8-O-β-D-glucuronide:quercetin is 10:1.0-10.0, preferably 10:1.0-6.0, and preferably It is preferably 10:0.1-0.4, and more preferably 10:0.1-0.3, and further preferably contains quercetin-3-O-robinobioside, where the ratio of quercetin-8-O-β-D-glucuronide:quercetin-3-O-robinobioside is 10:0.1-2.5 or 10:0.1-0.9, and preferably 10:0.15-0.5.

[0013] In a second aspect, the present invention provides a flavonoid active site of Abelmoschus maniihot flower containing flavonoid components in the following mass ratios, wherein the mass ratios of isoquercitrin:quercetin:quercetin-3'-O-glucoside:myricetin:quercetin-8-O-β-D-glucuronide:hypericin 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 effective portion of the Abelmoschus maniihot flower contains flavonoid components in the following mass ratio, and the mass ratio of isoquercitrin: quercetin: quercetin-3'-O-glucoside: myricetin: quercetin-8-O-β-D-glucuronide: hypericin is 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, then 0.8~1.2:0.2~0.8:0.8~2.1:0.18~0.6:0.8~2.0:0.7~2.0, then 0.8~1.2:0.2~0.7:0.8~1.6:0.2~0.5:0.8~1.8:0.8~1.4, then 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 effective portion of the Abelmoschus manihot flower contains flavonoid components in the following mass ratios of isoquercitrin:quercetin:quercetin-3'-O-glucoside:myricetin:quercetin-8-O-β-D-glucuronide:hypericin: 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 effective portion of the Abelmoschus maniihot flower contains flavonoid components in the following mass ratios of isoquercitrin:quercetin:quercetin-3'-O-glucoside:myricetin:quercetin-8-O-β-D-glucuronide:hypericin: 0.8-1.2:0.12-0.7:0.8-1.6:0.2-0.5:0.85-1.7:1.0-1.4.

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

[0018] More specifically, the flavonoid effective portion of Abelmoschus maniihot flower 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, and further 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, preferably 10:0.15-0.5.

[0019] More specifically, the flavonoid effective site of Abelmoschus maniihot flower contains quercetin, and the total content of 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%, preferably 69-82%, and further, the content of quercetin-3'-o-glucoside is 12% or more. 0.1-25%, and further the quercetin-3'-o-glucoside content is 12.6-23% or 13-20%, 13-20%, or 14-25%, or further the quercetin content is greater than 0.7%, further greater than 1.0%, further 1.0%-10%, further 1.5%-5%, or further the quercetin-8-O-β-D-glucuronide is 8.5-30%, further 12%-23%.

[0020] More specifically, the effective portion contains quercetin, and the total content of quercetin-3'-O-glucoside, myricetin, quercetin-8-O-β-D-glucuronide, isoquercitrin, rutin and hypericin is 55% or more, preferably 60% or more, preferably 65 to 85% or 66 to 84%, preferably 69 to 82%, or 69 to 90%, and further, the quercetin-3'-o-glucoside content is 12.1 to 25%, and further the quercetin-3'-o-glucoside content is 12.6 to 23% or 13 to 20%.

[0021] In a third aspect, the present invention provides a plant extract containing a flavonoid component, the mass contents of which are as follows: hypericin 10-25%, isoquercitrin 8-19%, quercetin-8-O-β-D-glucuronide 5-30%, quercetin-3'-o-glucoside 12.1-25%, further hypericin 12-23%, isoquercitrin 10-17%, quercetin-8-O-β-D-glucuronide 8.5-25%, quercetin-3'-o-glucoside 12.6-23%, further hypericin 13-22%, isoquercitrin 11-17%, quercetin-8-O-β-D-glucuronide 12-21%, quercetin-3'-o-glucoside 13-20%.

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

[0023] In a fourth aspect, the present invention provides an Abelmoschus maniifolia flower extract containing flavonoid components having the following mass contents: hypericin 8-26%, isoquercitrin 12.0-19.8%, quercetin-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 hypericin 11-22%, isoquercitrin 12.0-17%, quercetin-8-O-β-D-glucuronide 12-23%, myricetin 1.6-4.9% or 5.1-9.0%, quercetin-3'-o-glucoside 13-22%, quercetin 1.4-8% or 1.4-7.8%.

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

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

[0026] In a fifth aspect, the present invention provides a composition comprising flavonoid components in the following mass ratios, wherein the mass ratio of isoquercitrin:quercetin:quercetin-3'-O-glucoside:myricetin:quercetin-8-O-β-D-glucuronide:hypericin is 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, then 1: 0.16~1.0: 0.6~2.4: 0.14~0.75: 0.6~2.5: 0.6~2.4, then 1: 0.2~0.8: 0.8~2.1: 0.18~0.6: 0.8~2.0: 0.7~2.0, then 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 or 1.2:0.05~1.6:0.2~4.6:0.05~1.2:0.2~3.5:0.25~3.6, further 1:0.1~1.2:0.4~3.1:0.10~0.9:0.4~3.1:0.5~3.0, further 1:0.16~1.0:0.6~2 .4:0.14~0.75:0.6~2.5:0.6~2.4, then 1:0.2~0.8:0.8~2.1:0.18~0.6:0.8~2.0:0.7~2.0, then 1:0.2~0.7:0.8~1.6:0.2~0.5:0.8~1.8:0.8~1.4, then 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, and the mass ratio of the 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. Furthermore, the content of quercetin-3'-o-glucoside in the flavonoid component is 12.1-25%, and further the content of quercetin-3'-o-glucoside is 12.6-23% or 13-20%, and further contains 0.08-2.5% quercetin-3-O-robinobioside, preferably 0.1-1.5%, further 0.1-0.9%, and further preferably 0.1-0.5%.

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

[0029] In a sixth aspect, the present invention provides a pharmaceutical composition, the pharmaceutical composition comprising the above-mentioned effective part, the above-mentioned Abelmoschus manihot extract or the above-mentioned composition, the pharmaceutical composition further comprising a pharmaceutically acceptable carrier.

[0030] Specifically, the pharma- ceutically acceptable carrier includes a solvent, an emulsifier, a disintegrant, a filler, a solubilizer, an antioxidant, a pH adjuster, an osmotic pressure adjuster, a bacteriostat, a diluent, a lubricant, a binder and / or a film former, and the lubricant does not include magnesium stearate.

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

[0032] In a seventh aspect, the present invention provides a use of the above-mentioned effective site, the above-mentioned Abelmoschus manihot extract, the above-mentioned composition or the above-mentioned pharmaceutical composition in the preparation of a medicament for nephropathy, preferably the nephropathy is diabetic nephropathy or diabetic nephropathy or nephritis accompanied by renal fibrosis.

[0033] In an eighth aspect, the present invention provides a use of the above-mentioned effective part, the above-mentioned Abelmoschus manihot extract, the above-mentioned composition or the above-mentioned pharmaceutical composition in the preparation of a medicament for an ophthalmic disease, wherein the above-mentioned ophthalmic disease is preferably macular degeneration, visual fatigue or cataract, diabetic retinopathy, age-related macular degeneration, central exudative chorioretinitis, macular degeneration due to high myopia.

[0034] In a ninth aspect, the present invention provides a use of the above-mentioned effective site, the above-mentioned Abelmoschus manihot extract, the above-mentioned composition or the above-mentioned pharmaceutical composition in the preparation of a drug for treating lupus erythematosus nephritis, or contrast-induced nephropathy, or pulmonary fibrosis, or heart failure, or lowering uric acid, or an SGLT2 inhibitor.

[0035] In a tenth aspect, the present invention provides a method for preparing the above-mentioned effective part or the above-mentioned Abelmoscus mani hot extract, the method comprising the steps of (1) extracting the Abelmoscus mani hot flowers or the medicinal parts of Abelmoscus mani hot with ethanol to prepare an extract, (2) concentrating the extract and then extracting it to prepare an extract, and (3) removing the solvent from the extract, followed by elution with a macroporous resin to prepare a flavonoid effective part or extract of Abelmoscus mani hot flowers, and preferably the extraction method is percolation.

[0036] Specifically, the amount of ethanol used in step (1) is 10 to 25 times the amount of Abelmoschus manii flowers or the medicinal parts of Abelmoschus manii, the ethanol is a 60 to 95% ethanol solution, the extracting agent for the extract in step (2) is n-butanol, petroleum ether or ethyl acetate, the extracting method is continuous countercurrent extracting, the material liquid ratio of the extracting is 0.8 to 4:1, the number of extracting stages of the extracting 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 of the macroporous resin in step (3) is as follows: the diameter-to-height ratio of the macroporous resin is 1:4-1:9, the loading buffer concentration is 0.10-0.30 g raw material / mL, the loading buffer volume is 4-12 BV, loading is performed at a flow rate of 1-4 BV / h, impurities are removed using 4-8 BV of pure water and 1-5 BV of 3-15% ethanol at a flow rate of 0.5-4 BV / h, and elution is performed using 2-8 BV of 50-80% ethanol at a flow rate of 1-5 BV / h.

[0039] In an eleventh aspect, the present invention provides a method for preparing the above-mentioned effective part or the above-mentioned extract of Abelmoscus manihot, the preparation method including the steps of (1) extracting Abelmoscus manihot flowers or medicinal parts of Abelmoscus manihot with ethanol to prepare an extract, (2) adding a clarifying agent to the extract, treating with water bath, and filtering to remove the supernatant, and (3) eluting the supernatant with polyamide resin to prepare the flavonoid effective part or extract of Abelmoscus manihot flowers, and preferably the extraction method is percolation.

[0040] Specifically, the amount of ethanol used in step (1) is 10 to 25 times the amount of Abelmoschus manii flowers or Abelmoschus manii medicinal parts, 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 diameter to height ratio of the resin 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 aspect, the present invention provides a method for preparing the above-mentioned effective part or the above-mentioned extract of Abelmoscus mani hot, the preparation method including the steps of (1) extracting Abelmoscus mani hot flowers or medicinal parts of Abelmoscus mani hot with ethanol to prepare an extract, (2) adjusting the pH value of the extract to 2.0-3.0, refrigerating, filtering to remove precipitate, and adding water to dissolve, (3) extracting the solution to prepare an extract, and (4) removing the solvent from the extract and then eluting with polyamide resin to prepare a flavonoid effective part or extract of Abelmoscus mani hot flowers, and preferably the extraction method is reflux.

[0042] Specifically, the amount of ethanol used in step (1) is 10-25 times the amount of Abelmoschus manii flowers or Abelmoschus manii medicinal parts, the ethanol is a 60-95% 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 extracting agent for the extract in step (3) is n-butanol, petroleum ether or ethyl acetate, the extracting method is continuous countercurrent extracting, the material liquid ratio of the extracting is 0.8-4:1, the number of extracting stages of the extracting is 1-5, the diameter-to-height ratio of the resin in step (4) is 1:4-1:9, the loading buffer concentration is 0.10-0.60 g raw material / mL, the loading buffer volume is 4-12 BV, and elution is performed using 4-8 BV of pure water and 4-8 BV of 60-95% ethanol.

[0043] In addition to detecting seven components, namely hypericin, rutin, isoquercitrin, quercetin-8-O-β-D-glucuronide (hibifolin), myricetin, quercetin-3'-o-glucoside, and quercetin, the flavonoid effective site of Abelmoschus manihot flower described in the present invention also detects the seven components, namely hypericin, rutin, isoquercitrin, quercetin-8-O-β-D-glucuronide (hibifolin), myricetin, quercetin-3'-o-glucoside, during the detection process, the solid sample prepared in the embodiment of the present invention also simultaneously performs content conversion for the peak area proportional to the content of quercetin-3-O-robinobioside, content conversion calculation formula = hypericin control product peak area / hypericin control product concentration * correction factor * quercetin-3-O-robinobioside peak area * constant volume / weighing volume (after water removal), the content is 2.0% or less, more preferably 0.9% or less, and preferably 0.1% to 0.7%. The content of the total composition of the detected multiple flavonoids is 70% or more, more preferably 75% or more, and even more preferably 80% to 90%. Effect of the Invention

[0044] The present invention has the following advantages.

[0045] (1) The flavonoid effective portion or extract of Abelmoschus maniihot flower described in the present invention not only has the effect of treating various nephropathy such as diabetic nephropathy, contrast agent-induced nephropathy or lupus erythematosus nephritis, but also has the effect of treating ophthalmological diseases, particularly various diseases associated with blood flow obstruction and ocular vascular obstruction, such as age-related macular degeneration, central exudative chorioretinitis and macular degeneration due to severe myopia, and in particular, the flavonoid effective portion or extract of Abelmoschus maniihot flower described in the present invention also has the effect of treating pulmonary fibrosis.

[0046] (2) The preparation method described in the present invention has a simple process, a short production cycle, mild processing conditions, low energy consumption, and good isolation effect of the seven flavonoids from Abelmoschus maniifolia flower. The raw materials and reagents used are widely available, low cost, and easy to realize industrialized large-scale production.

[0047] (3) The flavonoid active part or extract or composition of Abelmoschus maniihot flower described in the present invention has fewer side effects and provides a better treatment option for patients. [Brief description of the drawings]

[0048] [Figure 1] 1 is a graph showing the pathological changes detected in rats after repeated administration of different doses for 3 months, where A is the normal group, B is the sample group of Example 9.2, C is the Abel Mosque capsule group, and D is the group of Example 2-A4. [Diagram 2] 1 is a graph showing the pathological results of the DKD group, the HK group, and the HT group. [Diagram 3] 1 is a graph showing the results of detecting SLC5A2 mRNA expression in the DKD, HK, and HT groups. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

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

[0050] The Abelmoschus manii hot flowers in the following examples are dried corollas of Abelmoschus manii hot and, unless otherwise stated, Abelmoschus manii hot flowers from the same batch.

[0051] The content of seven components was measured for the flavonoid effective part of Abelmoschus maniihot flowers prepared in each example. The seven components are hypericin, rutin, isoquercitrin, quercetin-8-O-β-D-glucuronide, myricetin, quercetin-3'-o-glucoside, and quercetin. During the preparation process of the effective part sample, the content of each component was also measured for the solution of the effective part before drying, and there was a slight change in the content of the sample before and after drying. The detection method was the UPLC method reported in "Measurement of Seven Components of Abelmoschus maniihot Flowers by One-Test-Multiple Evaluation Method" ("Drug Analysis Journal", 12th issue, 2013, 2082-2087), and the content of the components was measured.

[0052] The extract liquid ratio (ie extract:extract liquid) is a volume ratio.

[0053] Example 1. Preparation of flavonoid active site from Abelmoschus maniihot flower 100g of Abelmoschus manii flowers were crushed into coarse powder and extracted with 15 times 60% ethanol by percolation to obtain Abelmoschus manii flowers extract. The n-butanol was removed from the Abelmoschus manii flowers extract, diluted with water, and adsorbed with activated carbon, and then continuously countercurrent extracted with n-butanol. The ratio of the extract material liquid (i.e., extract:extract) was 1.5:1, the extract loading speed was 40mL / min, the rotation speed of the extract extractor was 40Hz, and the number of extract stages was 2. The extract liquid was decompressed at 40℃ to recover the solvent, and then treated with D101 macroporous resin to obtain macroporous resin. The fat treatment process is as follows: the diameter to height ratio of the macroporous resin is 1:6, the loading buffer (aqueous solution) concentration is 0.15g raw material / mL, the loading buffer volume is 7BV, loaded at a flow rate of 2BV / h, decontaminated sequentially with 6BV of pure water and 3BV of 5% ethanol at a flow rate of 2BV / h, and eluted with 4BV of 60% ethanol at a flow rate of 2BV / h to obtain an eluate, and then the eluate is decompressed under 50℃ to recover ethanol, and then the flavonoid extract of Abelmoschus manii flower is obtained, the content is measured, and the content of seven components is adjusted by the accession method according to the content measurement result. The content of each component is shown in the table below. The total solid content of the flavonoid effective part of Abelmoschus manii flower is 2.23g, and the seven components are a total of 1769mg, accounting for 79.32% of the total solid content.

[0054] [Table 1]

[0055] Example 2. Preparation of flavonoid active site from Abelmoschus maniihot flower Example 2-1 100g of Abelmoschus manii flowers are crushed into coarse powder and extracted with 18 times 70% ethanol by percolation to obtain Abelmoschus manii flowers extract. The ethanol is removed from the Abelmoschus manii flowers extract, diluted with water, and continuously countercurrent extracted with ethyl acetate. The ratio of the extract material liquid (i.e., extract agent:extract liquid) is 2.5:1, the extract loading speed is 80mL / min, the rotation speed of the extract extractor is 40Hz, the number of extract stages is 4, the extract liquid is decompressed at 40℃ to recover the solvent, and then treated with D101 macroporous resin. The macroporous resin treatment process The diameter to height ratio of macroporous resin is 1:5, loading buffer concentration is 0.15g raw material / mL, loading buffer volume is 6BV, loading at a flow rate of 2BV / h, removing impurities with 6BV of pure water and 3BV of 5% ethanol at a flow rate of 2BV / h, eluting with 4BV of 60% ethanol at a flow rate of 1.5BV / h to obtain an eluate, then decompressing the eluate at 60°C to recover ethanol, and obtaining an Abelmoschus manii flower flavonoid extract, measuring the content, and adjusting the content of seven components according to the content measurement result by the accession method. The content of each component is shown in the table below. The total solid content of the flavonoid effective part of Abelmoschus manii flower is 2.99. The content of quercetin-3-O-robinobioside is 0.28%.

[0056] [Table 2-1]

[0057] Example 2 - A. Preparation of the flavonoid active site of Abelmoschus manihot flower According to the method of Example 2 above, the Abelmoschus manii flower was enlarged 10 times, and another two batches of Abelmoschus manii flower flavonoid effective sites were prepared, and the samples in Table 2-2. Sample content measurement data were obtained. The quercetin-3-O-robinobioside content is 0.20-0.50%.

[0058] [Table 2-2]

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

[0060] [Table 2-3]

[0061] Example 2-A3 Hypericin, isoquercitrin, hibifolin, myricetin, quercetin-3'-o-glucoside, quercetin, and quercetin 3-O-robinobioside were taken as raw materials, and the ingredients 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), and the total flavonoid composition was obtained by preparation.

[0062] Example 2-A4 Hypericin, isoquercitrin, hibifolin, myricetin, quercetin-3'-o-glucoside, quercetin, and quercetin 3-O-robinobioside were taken as raw materials, and the ingredients 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), and the total flavonoid composition was obtained by preparation.

[0063] Example 2-B In order to prepare the effective part of Abelmoschus manii flower that meets the mass uniformity requirements, from the results of the content of each component of the extract of Example 2-1 of multiple batches, in order to investigate the relationship between the content of the drug and the above-mentioned components of the extract, three different batches of Abelmoschus manii flower with large component differences were mixed according to the software calculation method, and 2000g of Abelmoschus manii flower after mixing was taken and extracted with 18 times 70% ethanol by percolation, and the ethanol was removed from the Abelmoschus manii flower extract, diluted with water, and continuously countercurrent extracted with ethyl acetate, the extract material liquid ratio (i.e. extract:extract) was 3:1, the extract loading speed was 120mL / min, and the extract extraction machine was The rotation speed is 60Hz, the number of extraction stages is 4, the extract liquid is decompressed at 50℃ to recover the solvent, then diluted with water, centrifuged, the supernatant is taken and treated with D101 macroporous resin, the treatment process of the macroporous resin is as follows: the diameter to height ratio of the macroporous resin is 1:6, the loading buffer concentration is 0.15g raw material / mL, the loading buffer volume is 7BV, loaded at a flow rate of 2BV / h, 6BV of pure water and 3BV of 5% ethanol are used to remove impurities at a flow rate of 2BV / h, eluted with 4BV of 60% ethanol at a flow rate of 2BV / h to obtain the eluate, concentrated, decompressed and dried at 60℃ to obtain the total solid content. It contains 0.22% rutin, 20.58% hypericin, 17.81% isoquercitrin, 16.07% hibifolin, 5.15% myricetin, 17.37% quercetin-3'-o-glucoside, and 1.77% quercetin, and the peak area equivalent of quercetin-3-O-robinobioside is 0.43%. The mass ratio between each component is shown in Table 2-4 below.

[0064] [Table 2-4]

[0065] Example 3 Preparation of flavonoid active site from Abelmoschus manihot flower Example 3.1 100g of Abelmoschus manii flowers are crushed into coarse powder and extracted with 18 times 60% ethanol by percolation to obtain Abelmoschus manii flowers extract. The ethanol is removed from the Abelmoschus manii flowers extract, diluted with water, and continuously countercurrent extracted with ethyl acetate. The ratio of the extract material liquid (i.e., extract agent:extract liquid) is 3:1, the extract loading speed is 100mL / min, the rotation speed of the extract extractor is 40Hz, the number of extract stages is 5, the extract liquid is decompressed at 40℃ to recover the solvent, and then treated with D101 macroporous resin. The macroporous resin treatment process The process is as follows: the diameter to height ratio of the macroporous resin is 1:8, the loading buffer concentration is 0.15g raw material / mL, the loading buffer volume is 8BV, loading at a flow rate of 2BV / h, removing impurities with 6BV of pure water and 3BV of 5% ethanol at a flow rate of 2BV / h, eluting with 4BV of 60% ethanol at a flow rate of 3BV / h to obtain an eluate, then reducing the pressure of the eluate at 60°C to recover ethanol, and obtaining an Abelmoschus manii flower flavonoid extract, measuring the content, and adjusting the content of seven components by the accession method according to the content measurement result. The content of each component is shown in the table below. The total solid content of the flavonoid effective part of Abelmoschus manii flower is 3.18g. The content of quercetin-3-O-robinobioside is 0.20-0.50%.

[0066] [Table 3]

[0067] 3.2 According to the preparation method of Example 3.1, the experiment was repeated on 500g of Abelmoschus manii flower drug, and the content data of the two batches obtained are shown in Table 4 below. The content of quercetin-3-O-robinobioside is 0.30-0.80%. A indicates "each component / total solid content (%)", and B indicates "each component / isoquercitrin".

[0068] [Table 4]

[0069] Example 4 Preparation of flavonoid active site from Abelmoschus manihot flower Example 4-1 100g of Abelmoschus manii hot flowers are extracted with 18 times 70% ethanol by percolation to obtain Abelmoschus manii hot flowers extract, the ethanol is removed from the Abelmoschus manii hot flowers extract, the extract is diluted with water, and the extract is continuously countercurrent extracted with n-butanol, the extract material liquid ratio (i.e., extract:extract) is 3:1, the extract loading speed is 120mL / min, the rotation speed of the extract extractor is 60Hz, the number of extract stages is 5, the extract liquid is decompressed at 50℃ to recover the solvent, and then treated with D101 macroporous resin, and the macroporous resin treatment process is carried out. The procedure is as follows: the diameter to height ratio of macroporous resin is 1:6, the loading buffer concentration is 0.15g raw material / mL, the loading buffer volume is 7BV, loading at a flow rate of 2BV / h, removing impurities with 6BV of pure water and 3BV of 5% ethanol at a flow rate of 2BV / h, eluting with 4BV of 60% ethanol at a flow rate of 2BV / h to obtain an eluate, then the eluate is decompressed under 60℃ to recover ethanol, and then the Abelmoschus manii flower extract is obtained, the content is measured, and the content of seven components is adjusted according to the content measurement result by the accession method. The content of each component is shown in the table below. The total solid content of the flavonoid effective part of Abelmoschus manii flower is 2.05g.

[0070] [Table 5]

[0071] Example 4-2 Abelmoschus manii hot flowers were extracted by percolation with 18 times 70% ethanol, and the ethanol was removed from the extract until it was 0.5-1.0 g raw material / mL, and the added amount of ZTC1+1-II clarifier was 4% B component and 2% A component, respectively. The water bath temperature was 60°C, the water bath incubation time was 60 minutes, and the resin 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 raw material / mL, and the loading buffer volume was 8BV. The eluate was obtained by eluting with 5BV of pure water and 5BV of 80% ethanol, concentrated, dried under reduced pressure at 60°C, and 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 effective parts of Abelmoschus maniihot flowers was obtained, and the content of quercetin-3-O-robinobioside was 0.39%.

[0072] [Table 5-1]

[0073] Example 5. Preparation of flavonoid active site from Abelmoschus maniihot flower Example 5-1 100g of Abelmoschus manii hot flowers were extracted with 15 times 70% ethanol by percolation to obtain Abelmoschus manii hot flowers extract, the ethanol was removed from the Abelmoschus manii hot flowers extract, diluted with water, and continuously countercurrent extracted with n-butanol, the extract material liquid ratio (i.e. extract:extract) was 2.5:1, the extract loading speed was 80mL / min, the rotation speed of the extract extractor was 50Hz, the number of extract stages of the extract was 4, the extract liquid was reduced in pressure at 50℃ to recover the solvent, and then treated with D101 macroporous resin, and the macroporous resin The process is as follows: the diameter to height ratio of the macroporous resin is 1:6, the loading buffer concentration is 0.2g raw material / mL, the loading buffer volume is 6BV, loading at a flow rate of 2BV / h, removing impurities with 1BV of pure water and 4BV of 5% ethanol at a flow rate of 2BV / h, eluting with 3BV of 60% ethanol at a flow rate of 2BV / h to obtain an eluate, decompressing the eluate at 60°C to recover ethanol, and then obtaining an Abelmoschus manii flower extract, measuring the content, and adjusting the content of seven components according to the content measurement results by the accession method. The content of each component is shown in the table below. The total solid content of the flavonoid effective part of Abelmoschus manii flower is 2.86g.

[0074] Example 5-2 Abelmoschus manii flower was refluxed and extracted with 15 times 70% ethanol, the ethanol was removed from the extract until the extract was 0.5-1.0g raw material / mL, the pH value was adjusted to 2.0-3.0 by adding 10% hydrochloric acid solution, refrigerated, filtered, washed, precipitated, dissolved by adding water, and extracted with ethyl acetate continuously countercurrently, the ratio of the raw material liquid of the extract (i.e., extract:extract) was 3:1, the number of the extracts of the extract was 4 stages, the extract liquid was reduced pressure under 50℃ to recover the solvent, and then diluted with water and treated with polyamide resin, the resin treatment process is as follows: the diameter to height ratio of the resin was 1:6, the loading buffer concentration was 0.15-0.5g raw material / mL, the loading buffer volume was 6BV, and the eluate was obtained by eluting with 4BV of pure water and 4BV of 70% ethanol, concentrated, and dried under reduced pressure at 60℃ to obtain the eluate. The quercetin-3-O-robinobioside content is 0.47%.

[0075] [Table 5-2]

[0076] Example 6. Preparation of flavonoid active site from Abelmoschus maniihot flower 100g of Abelmoschus manii hot flowers were extracted with 16 times 60% ethanol by percolation to obtain Abelmoschus manii hot flowers extract, the ethanol was removed from the Abelmoschus manii hot flowers extract, diluted with water, and continuously countercurrent extracted with ethyl acetate, the extract material liquid ratio (i.e. extract:extract) was 2:1, the extract loading speed was 100mL / min, the rotation speed of the extract extractor was 40Hz, the number of extract stages of the extract was 4, the extract liquid was decompressed under 50℃ to recover the solvent, and then treated with D101 macroporous resin, the treatment process of the macroporous resin was as follows: the diameter-to-height ratio of the macroporous resin was 100mL / min, ... The ratio is 1:6, the loading buffer concentration is 0.3g raw material / mL, the loading buffer volume is 3BV, the loading is performed at a flow rate of 2BV / h, impurities are removed using 3BV of pure water and 3BV of 5% ethanol at a flow rate of 2BV / h, and the eluate is obtained by eluting with 4BV of 70% ethanol at a flow rate of 2BV / h, the content is measured, and the content of the seven components is adjusted by adding seven specific components, the content of each component is shown in the following table, and then the eluate is decompressed under 60°C to recover ethanol, and the Abelmoschus manii flower extract is obtained, the content is measured, and the content of the seven components is adjusted according to the content measurement result by the accession method. The amount of addition is less than 10% of the total weight. The content of each component is shown in the following table. The total solid content of the flavonoid effective part of Abelmoschus manii flower is 3.16g.

[0077] [Table 6]

[0078] Example 7. Preparation of flavonoid active site of Abelmoschus maniihot flower (Comparative Example) Example 7-1 500g of Abelmoschus manii flower was crushed into coarse powder, extracted with 20 times 70% ethanol by percolation to obtain Abelmoschus manii flower extract, and then treated with D101 macroporous resin after decompression and solvent recovery. The treatment process of the macroporous resin is as follows: the diameter to height ratio of the macroporous resin is 1:4, the loading buffer concentration is 0.15g raw material / mL, the loading buffer volume is 6BV, and the loading is performed at a flow rate of 2BV / h. Impurities are removed using 7BV of pure water and 4BV of 5% ethanol at a flow rate of 2BV / h. The eluate is eluted at a flow rate of 1.5BV / h using 60% ethanol to obtain an eluate. The eluate is decompressed at 60℃ to recover ethanol, and the total solid content of Abelmoschus manii flower is 16.12g. The content of each component is shown in the table below.

[0079] [Table 7-1]

[0080] Example 7-2 100g of Abelmoschus manii flower was crushed into coarse powder and extracted with 18 times 70% ethanol by percolation to obtain Abelmoschus manii flower extract. After reducing pressure to recover the solvent, the extract was treated with D101 macroporous resin. The treatment process of the macroporous resin 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, and the loading was performed at a flow rate of 2BV / h. Impurities were removed using 6BV of pure water and 3BV of 5% ethanol at a flow rate of 2BV / h. The eluate was eluted at a flow rate of 1.5BV / h using 4BV of 60% ethanol. The eluate was reduced pressure at 60℃ to recover the ethanol, and the total solid content of Abelmoschus manii flower was 3.56g.

[0081] [Table 7-2]

[0082] Example 8. Preparation of flavonoid active site of Abelmoschus manihot flower (Comparative Example) 100g of Abelmoschus manii flowers were crushed into coarse powder and extracted using 18 times 70% ethanol percolation to obtain Abelmoschus manii flowers extract, the ethanol was removed from the Abelmoschus manii flowers extract, diluted with water, and continuously countercurrent extracted with ethyl acetate, the extract material liquid ratio (i.e. extract:extract) was 2.5:1, the extract loading speed was 80mL / min, the rotation speed of the extract extractor was 40Hz, the number of extract stages was 4, and the extract liquid was decompressed at 40℃ and the solvent was recovered. , treated with D101 macroporous resin, the treatment process of the macroporous resin is as follows: the diameter to height ratio of the macroporous resin is 1:5, the loading buffer concentration is 0.15g raw material / mL, the loading buffer volume is 6BV, loaded at a flow rate of 2BV / h, impurities are removed using 6BV of pure water and 3BV of 5% ethanol at a flow rate of 2BV / h, eluted with 4BV of 60% ethanol at a flow rate of 1.5BV / h to obtain an eluate, the content of which is measured, and then rutin and hypericin are added to the eluate. The content of each component is shown in the table below. After that, the pressure is reduced at 60℃ to recover the ethanol, and 4.28g of Abelmoschus manii hot flower total solids are obtained.

[0083] [Table 8]

[0084] Example 9. 9.1. Preparation of flavonoid active site from Abelmoschus maniihot flower 25 kg of Abelmoschus manii flowers were crushed into coarse powder and extracted using 18 times 70% ethanol percolation to obtain Abelmoschus manii flowers extract. The ethanol was removed from the Abelmoschus manii flowers extract, diluted with water, and continuously countercurrent extracted with ethyl acetate. The ratio of the extract material liquid (i.e., extract:extract) was 2.5:1, and the number of extract stages was 4. The extract liquid was decompressed under 40℃ to recover the solvent, and then treated with D101 macroporous resin. The treatment process of the macroporous resin was as follows: the diameter of the macroporous resin was 1.2 mm, and the diameter of the macroporous resin was 1.2 mm. The height ratio is 1:5, the loading buffer concentration is 0.15g raw material / mL, the loading buffer volume is 3BV, the loading is performed at a flow rate of 1.5BV / h, impurities are removed using 6BV of pure water and 3BV of 10% ethanol at a flow rate of 2BV / h, and elution is performed using 4BV of 60% ethanol at a flow rate of 1.5BV / h to obtain an eluate, and the eluate is decompressed at 60°C to recover ethanol, and then the total solid content of Abelmoschus manii flower is obtained, and the seven specific components are added to adjust the seven components (the content of the added components is 20% or less of the total weight). The content of each component is shown in the table below. The total solid content of the flavonoid effective part of Abelmoschus manii flower is obtained. The content of quercetin-3-O-robinobioside is 0.20-0.50%.

[0085] [Table 9]

[0086] 9.2. Preparation of flavonoid active site from Abelmoschus maniihot flower Referring to Example 2-B, 30 kg of Abelmoschus manii flower mixture is crushed into coarse powder, and extracted with 18 times 70% ethanol by percolation to obtain Abelmoschus manii flower extract, the ethanol is removed from the Abelmoschus manii flower extract, diluted with water, and continuously countercurrent extracted with ethyl acetate, the extract material liquid ratio (i.e. extract:extract) is 2.5:1, the extract loading speed is 80mL / min, the rotation speed of the extract extractor is 40Hz, the number of extract stages is 4, the extract liquid is decompressed under 40℃ to recover the solvent, and then treated with D101 macroporous resin. The process of the macroporous resin is as follows: the diameter to height ratio of the macroporous resin is 1:5, the loading buffer concentration is 0.15g raw material / mL, the loading buffer volume is 5BV, loading at a flow rate of 1.5BV / h, removing impurities with 3BV of pure water and 4BV of 5% ethanol at a flow rate of 2BV / h, eluting with 5BV of 60% ethanol at a flow rate of 1.5BV / h to obtain the eluate, decompressing the eluate at 65℃ to recover the ethanol, and then obtaining the total solids of Abelmoschus manii flower, which are vacuum dried and crushed, and the flavonoid content (%) of the seven components is detected. The results are shown below, where the content of quercetin-3-O-robinobioside is 0.20-0.50%.

[0087] [Table 10]

[0088] Example 11. Preparation of eye drops containing flavonoids from Abelmoschus maniihot flowers Preparation process: Prepare the active site according to any one of the methods of Examples 1 to 7, add water for injection, dissolve by stirring, adjust the volume to 100 L, adjust the pH to 7, filter, fill, and sterilize to obtain eye drops. The pH adjuster used is sodium hydroxide and / or hydrochloric acid.

[0089] Example 12. Pharmacological experiments in glucose 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 the experimental mice were acclimated for 3 days, they were fed a high-fat, high-carbohydrate diet (homemade in the laboratory, recipe: lard: sucrose: egg yolk: basal diet = 18: 20: 3: 59) for 8 weeks, and then STZ sodium citrate solution was intraperitoneally injected at 100 mg / kg for 7 days. On the 8th day, the blood glucose level of the mice was detected through the tail vein, and the blood glucose level exceeding 16.7 mmol / L was used as the criterion for forming a diabetic model. When the blood glucose level exceeded 13.8 mmol / L, urinary protein appeared, and renal function became abnormal, it indicated the success of the modeling. Experimental grouping: normal group, model group, positive drug group (dagliflozin tablet, 45.5 mg / kg), sample group prepared by the method of Example 2-1 (62.4 mg / kg), sample group prepared by the method of Example 6 (62.4 mg / kg), sample group prepared by the method of Example 8 (62.4 mg / kg), total of 6 groups, 15 mice per group, urinary protein and urinary creatinine were detected after 4 weeks of administration, and daily intake and mouse condition were calculated. The detection results are shown in Table 12-1 below.

[0090] [Table 11]

[0091] Each group of the examples has the effect of reducing urinary protein, and the sample group of Example 2-1 can significantly reduce urinary protein and urinary albumin / urinary creatinine ratio (ACR). As can be seen from the observation results of the mouse condition, the number of mice that suffered from diarrhea and bloating in the sample group of Example 2 was significantly less than that of 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 raising db / db mice for 17 weeks, urinary protein was detected, and when urinary protein appeared and renal function became abnormal, the model was successful and was used for further 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), with a total of 6 groups, 15 mice per group, and 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 of the examples has the effect of reducing urinary protein, and each sample group can reduce urinary protein and urinary albumin / urinary creatinine ratio (ACR). As can be seen from the observation results of the mouse condition, the number of mice that suffered from diarrhea and bloating in the sample groups of Example 3-1 and Example 9-1 was significantly less than that of the other groups.

[0095] Example 13. Effects of flavonoids on ocular blood flow and choroidal neovascularization Experimental animals: rats, weighing 150-180g, SPF grade, half male and half female. Method of administration: Oral administration Experimental modeling: Healthy male Brown-Norway rats were selected, and the animals were anesthetized by intramuscular injection of ketamine (50 mg / kg). The pupils were dilated with a mixture of 0.5% tropicamide and neoforin. Using a krypton yellow laser (Kojima Co., Ltd., USA) (wavelength 568 nm, parameters: spot diameter 100 μm, exposure time 0.1 s, output 150-200 mW), 8 points of photocoagulation were performed on the retina between the optic disc, 2-4 disc diameters from the optic disc, around the optic disc, and between the large blood vessels under a 120D dedicated front lens. If the Bruch's membrane was broken and small bubbles occurred, it was considered a qualified photocoagulation point. Retinal, choroidal, and vitreous hemorrhage were excluded. Experimental Grouping: Drug grouping and dosage: normal group, model group, positive control drug group (bitexofen 1.35mg / kg), Example 2-1 sample group (69.9mg / kg), Example 3.1 sample group (87.36mg / kg), Example 7 sample group (87.36mg / kg), Example 9.1 sample group (87.36mg / kg), Example 9.2 sample group (87.36mg / kg, solid content prepared according to Example 9.2), each group was made up of 15 mice, and the ocular choroidal blood flow inhibition rate and choroidal neovascular area were detected after 4 weeks of administration. The detection results are shown in Table 13 below.

[0096] [Table 13]

[0097] Conclusion: The samples of each Example have an inhibitory effect on choroidal blood flow inhibition rate and choroidal neovascularization area, and the Example 2 sample group, the Example 3 sample group, the Example 9.1 sample, and the Example 9.2 sample can significantly inhibit choroidal neovascularization.

[0098] Example 14. Effects of flavonoids on contrast-induced renal injury Experimental animals: SD rats, weighing 200-250g, SPF grade, half male and half female. Method of administration: Oral administration Experimental modeling: Healthy male SD rats were selected and first, furosemide (10ml / kg) was injected intramuscularly, followed by tail vein injections of a prostaglandin synthesis inhibitor (indomethacin, 10mg / kg, dissolved in dimethylsulfoxide and diluted in 1.25% NaHCO3 solution) and a nitric oxide synthase inhibitor (N-nitro-L-arginine methyl ester, L-NAME, 10mg / kg, dissolved in 0.9% NaCl solution) and 76% pantethine (10ml / kg) every 15 minutes. Modeling success criteria The diagnostic criteria for acute kidney injury recommended in the 2012 KDIGOAKI clinical guidelines (a diagnosis was made if any of the following was met): (1) A rise in serum creatinine of more than 26.5 μmol / L (0.3 mg / dl) within 48 hours, (2) A rise in serum creatinine of more than 50% of baseline confirmed or estimated to have occurred within 7 days, (3) A decrease in urine output of <0.5 ml / (kg*h) lasting for more than 6 hours. Here, in animal models, a rise in serum creatinine of more than 50% of baseline is often used as a diagnostic criterion for acute kidney injury, i.e., a modeling success criterion. Drug grouping and dosage: normal group, model group, positive control drug group (Irbesarta 50mg / kg / d), Example 2-1 sample group (69.9mg / kg), Example 3.1 sample group (87.36mg / kg), Example 7 sample group (87.36mg / kg), Example 9.1 sample group (87.36mg / kg), Example 9.2 sample group (87.36mg / kg, solid content prepared in Example 9.2) were used with 15 mice per group, and the ocular choroidal blood flow inhibition rate and choroidal neovascular area were detected after 4 weeks of administration. The detection results are shown in Table 14 below.

[0099] [Table 14]

[0100] Conclusion: The serum creatinine and serum urea nitrogen of rats with contrast agent-induced kidney damage in the model group were significantly higher than those of each treatment group, and serum creatinine and serum urea nitrogen were all reduced to different degrees in the Example 9.1 sample group, Example 2-A1 sample group, Example 3.1 sample, Example 8 sample and Example 6 sample.

[0101] Example 15. Effects of flavonoids on lupus erythematosus nephritis Twenty-six male MRL / lpr mice (lupus erythematosus nephritis mice) were selected as one group for observation, weighing 18-22g, aged 13 weeks, and purchased from Nanjing Hualianke Biotechnology Co., Ltd. Twenty-four male C57BL / 6 mice were selected as the normal group, weighing 18-22g, aged 13 weeks, and purchased from Wuhan Hualianke Biotechnology Co., Ltd. Drug grouping and dosage: normal group, MRL / lpr mouse model group, positive control drug 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), Example 8 sample group (87.36 mg / kg, solid content prepared in Example 9.2), 15 mice per group, were administered for 4 weeks. Experimental Indicators: After the administration, urine was collected for 24 hours to detect urinary protein. After collecting urine, the eyes of all mice were removed and blood was taken, centrifuged to obtain serum, which was used for detecting antinuclear antibodies. At the same time, the kidneys were removed and quickly frozen in liquid nitrogen, then transferred to a refrigerator at minus 80 degrees for storage, and used for kidney immunoassay detection. The detection results are shown in Table 15 below.

[0102] [Table 15]

[0103] Conclusion: The urinary protein, serum antinuclear antibody levels, and IgG and C3 deposition in renal tissue of lupus erythematosus nephritis mice in the model group were significantly higher than those in each treatment group, and the Example 9.1 sample group, Example 2-B sample group, Example 3.1 sample, Example 8 sample and Example 7 sample were all able to improve urinary protein, serum antinuclear antibody levels, and IgG and C3 deposition in renal tissue, with the Example 2-B sample, Example 3.1 sample and Example 5-2 sample being more effective.

[0104] Example 16. Toxic Effects of Flavonoids Experimental animals: rats, weighing 150-180g, SPF grade, half male and half female. Method of administration: Oral administration Drug grouping and dosage: normal group, Abel Mosque capsule group (extract 8g / kg), Example 2-A1 sample group (5g / kg), Example 9.1 sample group (8g / kg), 30 animals in each group, administered once a day for 12 consecutive weeks, dosage calculated according to the extract.

[0105] Test Results: After the end of administration, the kidney weight, kidney volume and organ index of the Abelmosque capsule group increased compared to the normal group, and pathological examination showed moderate renal tubule hypertrophy in the Abelmosque capsule group, but no renal tubule hypertrophy was found in the Example 9.2 sample group and Example 2-A1 group by pathological detection. The detection results are shown in Table 16 and Figure 1 below.

[0106] [Table 16]

[0107] Example 17 Effect of total flavonoid effective sites on idiopathic pulmonary fibrosis Experimental animals: 60 Kunming mice, half male and half female, weighing 18-22 kg, clean grade. Animal grouping: The animals were randomly divided into six groups, namely, a normal group, a model group, a control drug 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 Abel Mosque capsule sample group (180 mg / kg in terms of extract), with 10 mice in each group. Experimental modeling: After three days of acclimation, the experimental mice were anesthetized by intraperitoneal injection of 4% chloral hydrate (0.01ml / g). They were fixed in a supine position and disinfected with routine methods. The midline of the neck was incised and the trachea was exposed by blunt dissection. The model group, control drug group, and treatment group were slowly injected with bleomycin (5mg / kg) through tracheal cartilage ring gap puncture, and the normal control group was injected with the same amount of saline. Immediately after drug injection, the mice were rotated in an upright position for 3-5 minutes to ensure that the drug solution was evenly distributed in both lungs. The skin was sutured, the sutured area was disinfected, and the mice were awakened and kept in a clean grade observation room. Administration method: Administration began on the second day of modeling. The treatment group was administered the above-mentioned dosage once a day, while the normal group and model group were administered the same amount of distilled water (10 ml / kg) in the same manner, for 28 consecutive days. Lung histopathological section staining: After administration, the right lung was excised, fixed, embedded, sectioned, and stained with HE according to the usual pathological method. The severity 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 can be seen from the experimental results, the degree of lung tissue alveolitis and pulmonary fibrosis in the model group of mice increased significantly, showing typical parenchymal lesions of pulmonary fibrosis. After 28 days of administration, the lung coefficient of the mice in each dosage group was significantly reduced, and the degree of lung tissue alveolitis and pulmonary fibrosis was significantly reduced. The total flavonoids effective site significantly reduced the degree of alveolitis and pulmonary fibrosis in pulmonary fibrosis mice, and had the effect of protecting the lung tissue of pulmonary fibrosis mice, and could reduce the damage to the lung tissue of fibrosis opponents.

[0111] Example 18 SGLT2 mechanism of action study Sodium-glucose contransporter2 (SLC5A2, also known as SGLT2) is a gene encoding a sodium-glucose cotransporter that reabsorbs more than 90% of glucose filtered in the glomerulus. SLC5A2 is expressed only in the brush border in the proximal tubule 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 SLC5A2 protein in renal tissue of patients with diabetes and diabetic nephropathy can play a role in the excretion of sodium and glucose, and therefore can protect renal tissue of patients with diabetes and diabetic nephropathy. Animal model: db / db mice, 18 weeks old Method of administration: Oral administration Experimental grouping: db / db mice with UACR>200mg / g were divided into three groups: diabetic nephropathy group (DKD), Abelmosque capsule group (DKD+HK, 0.84g / kg / d) and total flavonoid group (DKD+HT, sample of Example 2-A2, 0.0755g / kg / d). The Abelmosque capsule group and total flavonoid group were administered for four weeks (28d). Enzyme-linked immunosorbent assay (ELISA) was used to detect microalbumin (UACR) in urinary fluid of db / db mice, reverse transcription polymerase chain reaction (RT-PCR) was used to detect SLC5A2 mRNA expression level, and immunohistochemistry (IHC) was used to detect SLC5A2 protein expression level.

[0112] Test Results: Compared to the DKD group, there was no significant difference in the weight before grouping DKD (N=5), HK (N=5), HT (N=6), weight after 1 week administration and weight after 4 weeks administration DKD (N=5), HK (N=5), HT (N=6) in the HK group (Abel Mosque Capsule group) and HT group (total flavonoid group). There was no significant difference in blood glucose after 1 week administration and blood glucose after 4 weeks administration DKD (N=5), HK (N=5), HT (N=6). There was no significant difference in UACR value DKD (N=5), HK (N=5), HT (N=6) after 1 week administration. After 4 weeks administration, the HT group showed a significant decrease in UACR value compared to the DKD group (*P<0.05), and there was a significant difference. There was no significant difference in renal index DKD (N=5), HK (N=5), HT (N=6). Renal index was significantly increased in DKD (N=5), HK (N=5), and HT (N=6), *P<0.05, and there was a significant difference. As shown in Figure 2, in the DKD group, some glomeruli were lobulated, the basement membrane was significantly thickened, and the capillaries of the glomeruli were significantly compressed. In the mesangial area, mesangial cell proliferation was observed, and the mesangial interstitium was obviously expanded. In the HK group, the thickening of the glomerular basement membrane was significantly improved, the glomerular capillaries were 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 interstitium did not proliferate, and the proliferation of mesangial cells and mesangial matrix did not increase. As shown in Figure 3, the quantitative results by IHC-P verified that the expression level of SLC5A2 protein was significantly decreased in the Abel Mosque HK group (N = 5) and the HT group (N = 6) compared to the DKD group (N = 5). The RT-PCR results and the immunohistochemistry results showed that the mRNA expression level of SLC5A2 was significantly decreased in the Abel Mosque HK group (N = 5) and the HT group (N = 6) compared to the DKD group (N = 5). The ratio of SLC5A2 IHC-P quantitative expression amount to the number of glomeruli was significantly decreased in the Abel Mosque HK group (N = 5) and the HT group (N = 6) compared to the DKD group (N = 5), which was consistent with the quantitative results of IHC-P.

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

[0114] The above examples are merely specific and detailed representations of some embodiments of the present invention, and are not to be construed as limiting the patent scope of the present invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and all such modifications and improvements are within the scope of protection of the present invention. Therefore, the scope of protection of the patent of the present invention shall be subject to the scope of the attached claims.

[0115] (Additional Note) (Appendix 1) 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 site of Abelmoschus manihot flower is characterized by the above.

[0116] (Appendix 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, preferably 10:6.0-15:5.0-13:2.0-5.0:8.0-14, Further, the composition contains quercetin, and the ratio of quercetin-8-O-β-D-glucuronide:quercetin is 10:1.0 to 10.0, preferably 10:1.0 to 6.0, more preferably 10:1.5 to 5.0; The composition further contains rutin, and the ratio of quercetin-8-O-β-D-glucuronide to rutin is 10:0.05 to 0.6, preferably 10:0.1 to 0.4, and more preferably 10:0.1 to 0.3. More preferably, it contains quercetin-3-O-robinobioside, and the ratio of quercetin-8-O-β-D-glucuronide:quercetin-3-O-robinobioside is 10:0.1 to 2.5, 10:0.1 to 0.9, and preferably 10:0.15 to 0.5. The effective portion of claim 1.

[0117] (Appendix 3) The following mass ratios [Table 18] A flavonoid effective site characterized by containing a flavonoid component.

[0118] (Appendix 4) The following mass ratios [Table 19] , preferably [Table 20] , more preferably [Table 21] , more preferably [Table 22] , more preferably [Table 23] ,or [Table 24] ,or [Table 25] The effective portion according to claim 3, characterized in that it contains a flavonoid component.

[0119] (Appendix 5) 6. The effective site according to any one of claims 3 to 5, wherein the mass ratio of isoquercitrin is 1.2 or 1.0.

[0120] (Appendix 6) further containing rutin, wherein 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; Further, it contains quercetin-3-O-robinobioside, and the mass ratio of isoquercitrin to quercetin-3-O-robinobioside is 10:0.1 to 2.5, 10:0.1 to 0.9, and preferably 10:0.15 to 0.5. 6. The effective site according to any one of claims 3 to 5.

[0121] (Appendix 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 to 85%, preferably 69 to 82%, Further, the quercetin-3'-o-glucoside content is 12.1-25%, and further, the quercetin-3'-o-glucoside content is 12.6-23%, or 13-20%, or 13-20%, or 14-25%, or further, the content of quercetin is greater than 0.7%, further greater than 1.0%, further between 1.0% and 10%, further between 1.5% and 5%; or still further, quercetin-8-O-β-D-glucuronide is 8.5% to 30%, or further, 12% to 23%. 6. The effective site according to any one of claims 1 to 5.

[0122] (Appendix 8) the total content of quercetin, quercetin-3'-O-glucoside, myricetin, quercetin-8-O-β-D-glucuronide, isoquercitrin, rutin and hypericin contained in the effective portion is 55% or more, preferably 60% or more, preferably 65 to 85% or 66 to 84%, preferably 69 to 82%, or 69 to 90%, Further, the quercetin-3'-o-glucoside content is 12.1-25%, and further, the quercetin-3'-o-glucoside content is 12.6-23% or 13-20%. The effective portion of claim 7.

[0123] (Appendix 9) A plant extract containing flavonoid components, the plant extract having the following mass contents of components: Hypericin 10-25%, isoquercitrin 8-19%, quercetin-8-O-β-D-glucuronide 5-30%, quercetin-3'-o-glucoside 12.1-25%, furthermore Hypericin 12-23%, isoquercitrin 10-17%, quercetin-8-O-β-D-glucuronide 8.5-25%, quercetin-3'-o-glucoside 12.6-23%, furthermore Hypericin 13-22%, isoquercitrin 11-17%, quercetin-8-O-β-D-glucuronide 12-21%, quercetin-3'-o-glucoside 13-20%, A plant extract characterized by:

[0124] (Appendix 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, further 1.5%-5%, further 2-11% myricetin, preferably 3-7% myricetin, preferably 3-5% myricetin, further 0.08-2.5% quercetin-3-O-robinobioside, preferably 0.1-1.5%, further 0.1-0.9%, further 0.1-0.5%, and further, that the plant is Abelmoschus maniihot or okra flower, preferably Abelmoschus maniihot flower.

[0125] (Appendix 11) Abelmoschus manii hot flower extract containing flavonoid components, comprising the following mass content of components: Hypericin 8-26%, isoquercitrin 12.0-19.8%, quercetin-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 furthermore hypericin 11-22%, isoquercitrin 12.0-17%, quercetin-8-O-β-D-glucuronide 12-23%, myricetin 1.6-4.9% or 5.1-9.0%, quercetin-3'-o-glucoside 13-22%, quercetin 1.4-8% or 1.4-7.8%. Abelmoschus manii hot flower extract characterized by:

[0126] (Appendix 12) Further containing rutin in a mass content of 0.01 to 1.0%, further 0.05 to 0.8%, further 0.09 to 0.8%, further 0.1 to 0.6%, Further, the composition contains 0.08 to 2.5% of quercetin-3-O-robinobioside, preferably 0.1 to 1.5%, more preferably 0.1 to 0.9%, and even more preferably 0.1 to 0.5%. 12. The Abelmoschus manii hot flower extract according to claim 11,

[0127] (Appendix 13) The Abelmoschus manii hot flower extract described in Appendix 12, characterized in that the mass content of flavonoid components in the Abelmoschus manii hot flower extract is 55% or more, preferably 60% or more, preferably 65 to 85% or 66 to 84%, preferably 69 to 82% or 69 to 90%.

[0128] (Appendix 14) Contains flavonoid components in the following mass ratios: The mass ratio of isoquercitrin:quercetin:quercetin-3'-O-glucoside:myricetin:quercetin-8-O-β-D-glucuronide:hypericin is 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, then 1:0.16~1.0:0.6~2.4:0.14~0.75:0.6~2.5:0.6~2.4, then 1:0.2~0.8:0.8~2.1:0.18~0.6:0.8~2.0:0.7~2.0, then 1:0.2~0.7:0.8~1.6:0.2~0.5:0.8~1.8:0.8~1.4, then 1:0.2~0.6:1 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, even 1: 0.1~1.2: 0.4~3.1: 0.10~0.9: 0.4~3.1: 0.5~3.0, even 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.

[0129] (Appendix 15) The composition further contains rutin, and the mass ratio of the 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, Furthermore, the content of quercetin-3'-o-glucoside in the flavonoid components is 12.1 to 25%, and further the content of quercetin-3'-o-glucoside is 12.6 to 23% or 13 to 20%, Further, the composition contains 0.08 to 2.5% of quercetin-3-O-robinobioside, preferably 0.1 to 1.5%, more preferably 0.1 to 0.9%, and even more preferably 0.1 to 0.5%. 15. The composition according to claim 14,

[0130] (Appendix 16) The composition described in 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 to 85%, and preferably 69 to 82%.

[0131] (Appendix 17) A pharmaceutical composition comprising an effective site described in any one of Appendices 1 to 8, an Abelmoschus manihot extract described in any one of Appendices 9 to 13, or a composition described in any one of Appendices 14 to 16, and further comprising a pharma- ceutically acceptable carrier.

[0132] (Appendix 18) 18. The pharmaceutical composition according to claim 17, wherein the pharma- ceutically acceptable carrier comprises a solvent, an emulsifier, a disintegrant, a filler, a solubilizer, an antioxidant, a pH adjuster, an osmolality adjuster, a bacteriostat, a diluent, a lubricant, a binder and / or a film former, and the lubricant does not comprise magnesium stearate.

[0133] (Appendix 19) The pharmaceutical composition according to claim 18, wherein the pharmaceutical composition is in the form of an injection, tablet, suppository, ointment, gel, pill, tablet, granule, capsule or preparation, suspension, or powder.

[0134] (Appendix 20) Use of an effective site described in any one of Appendices 1 to 8, an Abelmoschus manihot extract described in any one of Appendices 9 to 13, a composition described in any one of Appendices 14 to 16, or a pharmaceutical composition described in any one of Appendices 17 to 19 in the preparation of a drug for nephropathy, preferably diabetic nephropathy or diabetic nephropathy or nephritis accompanied by renal fibrosis.

[0135] (Appendix 21) Use of an effective site described in any one of Appendices 1 to 8, an Abelmoschus manihot extract described in any one of Appendices 9 to 13, a composition described in any one of Appendices 14 to 16, or a combination of drugs described in any one of Appendices 17 to 19 in the preparation of an ophthalmological disease drug, preferably for macular degeneration, visual fatigue or cataract, diabetic retinopathy, age-related macular degeneration, central exudative chorioretinitis, macular degeneration due to high myopia.

[0136] (Appendix 22) Use of an active site described in any one of Appendices 1 to 8, an Abelmoschus manihot extract described in any one of Appendices 9 to 13, a composition described in any one of Appendices 14 to 16, or a combination of drugs described in any one of Appendices 17 to 19 in the preparation of a drug for treating lupus erythematosus nephritis, or contrast-induced nephropathy, or pulmonary fibrosis, or heart failure, or for lowering uric acid.

[0137] (Appendix 23) A method for preparing the effective part according to any one of appendices 1 to 8 or the Abelmoschus maniihot extract according to any one of appendices 9 to 13, (1) extracting the flowers of Abelmoschus manii or the medicinal parts of Abelmoschus manii with ethanol to prepare an extract; (2) concentrating the extract and then extracting the extract to prepare an extract; (3) removing the solvent from the extract and then eluting it with a macroporous resin to prepare a flavonoid active part or extract of Abelmoschus maniihot flower; wherein preferably the extraction method is percolation.

[0138] (Appendix 24) The amount of ethanol used in step (1) is 10 to 25 times the amount of the medicinal site of the Abelmoschus mani hot flower or Abelmoschus mani hot, and the ethanol is a 60 to 95% ethanol solution; The extracting agent for the extract in step (2) is n-butanol, petroleum ether or ethyl acetate, the method of the extract is continuous countercurrent extract, the material liquid ratio of the extract is 0.8-4:1, and the number of extract stages of the extract is 1-5; The type of the macroporous resin in step (3) is D101, HPD100 or AB-8; 24. The method of claim 23,

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

[0140] (Appendix 26) The preparation method according to claim 25, characterized in that the elution process of the macroporous resin is as follows: diameter-to-height ratio of the macroporous resin is 1:4 to 1:9, loading buffer concentration is 0.10 to 0.30 g raw material / mL, loading buffer volume is 4 to 12 BV, loading at a flow rate of 1 to 4 BV / h, removing impurities 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 eluting with 2 to 8 BV of 50 to 80% ethanol at a flow rate of 1 to 5 BV / h.

[0141] (Appendix 27) A method for preparing the effective part according to any one of appendices 1 to 8 or the Abelmoschus maniihot extract according to any one of appendices 9 to 13, (1) extracting the flowers of Abelmoschus manii or the medicinal parts of Abelmoschus manii with ethanol to prepare an extract; (2) adding a clarifying agent to the extract, subjecting it to a water bath treatment, and filtering to remove the supernatant; (3) Eluting the supernatant with a polyamide resin to prepare a flavonoid active part or extract of Abelmoschus manihot flower. wherein preferably the extraction method is percolation.

[0142] (Appendix 28) The amount of ethanol used in step (1) is 10 to 25 times the amount of the medicinal site of the Abelmoschus mani hot flower or Abelmoschus mani hot, and the ethanol is a 60 to 95% ethanol solution; In step (2), the water bath temperature is 50 to 70° C., and the water bath time is 30 to 90 minutes. In step (3), the diameter-to-height ratio of the resin is 1:4-1:9, the loading buffer concentration is 0.10-0.60 g raw material / mL, the loading buffer volume is 4-12 BV, and elution is performed using 4-8 BV of pure water and 4-8 BV of 60-95% ethanol. 28. The method for preparing the compound according to claim 27,

[0143] (Appendix 29) A method for preparing the effective part according to any one of appendices 1 to 8 or the Abelmoschus maniihot extract according to any one of appendices 9 to 13, (1) extracting the flowers of Abelmoschus manii or the medicinal parts of Abelmoschus manii with ethanol to prepare an extract; (2) adjusting the pH value of the extract to 2.0-3.0, refrigerating, filtering to remove precipitate, and adding water to dissolve; (3) extracting the solution to prepare an extract; (4) removing the solvent from the extract and then eluting the extract with a polyamide resin to prepare a flavonoid active site or extract of Abelmoschus maniihot flower; and preferably the extraction method is reflux.

[0144] (Appendix 30) The amount of ethanol used in step (1) is 10 to 25 times the amount of Abelmoschus manii flower or Abelmoschus manii active 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 extracting agent for the extract in step (3) is n-butanol, petroleum ether or ethyl acetate, the method of the extract is continuous countercurrent extract, the material liquid ratio of the extract is 0.8-4:1, and the number of extract stages of the extract is 1-5; In step (4), the diameter-to-height ratio of the resin is 1:4-1:9, the loading buffer concentration is 0.10-0.60 g raw material / mL, the loading buffer volume is 4-12 BV, and elution is performed using 4-8 BV of pure water and 4-8 BV of 60-95% ethanol. 30. The method of claim 29,

Claims

1. 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 site of Abelmoschus manihot flower is characterized by the above.

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, preferably 10: 6.0-15: 5.0-13: 2.0-5.0: 8.0-14; Further containing quercetin, the ratio of quercetin-8-O-β-D-glucuronide:quercetin is 10:1.0 to 10.0, preferably 10:1.0 to 6.0, preferably 10:1.5 to 5.0; Furthermore, it further contains rutin, and the ratio of quercetin-8-O-β-D-glucuronide to rutin is 10:0.05 to 0.6, preferably 10:0.1 to 0.4, and more preferably 10:0.1 to 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 to 2.5, 10:0.1 to 0.9, and preferably 10:0.15 to 0.

5. The effective portion according to claim 1 .

3. The following mass ratios 【Table 1】 A flavonoid effective site characterized by containing a flavonoid component.

4. The following mass ratios 【Table 2】 , preferably 【Table 3】 , more preferably 【Table 4】 , more preferably 【Table 5】 , more preferably 【Table 6】 ,or 【Table 7】 ,or 【Table 8】 The effective portion according to claim 3, characterized in that it contains flavonoid components.

5. The effective site according to any one of claims 3 to 5, characterized in that the mass ratio value of isoquercitrin is 1.2 or 1.

0.

6. further containing rutin, wherein 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; Further, 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 portion according to any one of claims 3 to 5.

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 to 85%, preferably 69 to 82%, Further, the quercetin-3'-o-glucoside content is 12.1-25%, and further, the quercetin-3'-o-glucoside content is 12.6-23%, or 13-20%, 13-20%, or 14-25%, or further, the quercetin content is greater than 0.7%, further greater than 1.0%, further between 1.0% and 10%, further between 1.5% and 5%; or still further, quercetin-8-O-β-D-glucuronide is 8.5-30%, further still, 12%-23%. The effective portion according to any one of claims 1 to 5.

8. The total content of quercetin, quercetin-3'-O-glucoside, myricetin, quercetin-8-O-β-D-glucuronide, isoquercitrin, rutin and hypericin contained in the effective portion is 55% or more, preferably 60% or more, preferably 65 to 85% or 66 to 84%, preferably 69 to 82%, or 69 to 90%, Further, the quercetin-3'-o-glucoside content is 12.1-25%, and further, the quercetin-3'-o-glucoside content is 12.6-23% or 13-20%. The effective portion according to claim 7.

9. A plant extract containing flavonoid components, the plant extract having the following mass contents of components: hypericin 10-25%, isoquercitrin 8-19%, quercetin-8-O-β-D-glucuronide 5-30%, quercetin-3'-o-glucoside 12.1-25%, further hypericin 12-23%, isoquercitrin 10-17%, quercetin-8-O-β-D-glucuronide 8.5-25%, quercetin-3'-o-glucoside 12.6-23%, further hypericin 13-22%, isoquercitrin 11-17%, quercetin-8-O-β-D-glucuronide 12-21%, quercetin-3'-o-glucoside 13-20%. A plant extract characterized by:

10. 10. The extract according to claim 9, characterized in that it contains 2-10% quercetin, preferably 2.5-9% quercetin, preferably 3-8.5% quercetin, even 1.5%-5%, and even further contains 2-11% myricetin, preferably 3-7% myricetin, preferably 3-5% myricetin, and further contains 0.08-2.5% quercetin-3-O-robinobioside, preferably 0.1-1.5%, even 0.1-0.9%, even more preferably 0.1-0.5%, and further wherein the plant is Abelmoschus maniihot or Okra flower, preferably Abelmoschus maniihot flower.

11. Abelmoschus manii hot flower extract containing flavonoid components, comprising the following mass content of components: Hypericin 8-26%, isoquercitrin 12.0-19.8%, quercetin-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, hypericin 11-22%, isoquercitrin 12.0-17%, quercetin-8-O-β-D-glucuronide 12-23%, myricetin 1.6-4.9% or 5.1-9.0%, quercetin-3'-o-glucoside 13-22%, quercetin 1.4-8% or 1.4-7.8%. Abelmoschus manii hot flower extract characterized by:

12. further comprising rutin in a mass content of 0.01-1.0%, further 0.05-0.8%, further 0.09-0.8%, further 0.1-0.6%, Further, it contains 0.08 to 2.5% of quercetin-3-O-robinobioside, preferably 0.1 to 1.5%, more preferably 0.1 to 0.9%, and even more preferably 0.1 to 0.5%. The Abelmoschus manii hot flower extract according to claim 11.

13. The Abelmoschus manii hot flower extract according to claim 12, characterized in that the mass content of flavonoid components in the Abelmoschus manii hot flower extract is 55% or more, preferably 60% or more, preferably 65-85% or 66-84%, preferably 69-82% or 69-90%.

14. Contains flavonoid components in the following mass ratios: The mass ratio of isoquercitrin:quercetin:quercetin-3'-O-glucoside:myricetin:quercetin-8-O-β-D-glucuronide:hypericin is 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 to 1.2: 0.2 to 0.4: 0.9 to 1.7: 1.0 to 1.3 or 1.2: 0.05 to 1.6: 0.2 to 4.6: 0.05 to 1.2: 0.2 to 3.5: 0.25 to 3.6, further 1: 0.1 to 1.2: 0.4 to 3.1: 0.10 to 0.9: 0.4 to 3.1: 0.5 to 3.0, further 1: 0.16 to 1.0: 0.6 to 2.4: 0.14 to 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.

15. The composition further contains rutin, and the mass ratio of the 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; Furthermore, the content of quercetin-3'-o-glucoside in the flavonoid component is 12.1 to 25%, and further the content of quercetin-3'-o-glucoside is 12.6 to 23% or 13 to 20%, Further, it contains 0.08 to 2.5% of quercetin-3-O-robinobioside, preferably 0.1 to 1.5%, more preferably 0.1 to 0.9%, and even more preferably 0.1 to 0.5%.

15. The composition of claim 14.

16. The composition according to claim 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 to 85%, preferably 69 to 82%.

17. A pharmaceutical composition comprising an effective portion according to any one of claims 1 to 8, an Abelmoschus manihot extract according to any one of claims 9 to 13, or a composition according to any one of claims 14 to 16, and further comprising a pharma- ceutically acceptable carrier.

18. 18. The pharmaceutical composition of claim 17, wherein the pharma- ceutically acceptable carrier comprises a solvent, an emulsifier, a disintegrant, a filler, a solubilizer, an antioxidant, a pH adjuster, an osmolality adjuster, a bacteriostat, a diluent, a lubricant, a binder and / or a film former, wherein the lubricant does not comprise magnesium stearate.

19. The pharmaceutical composition according to claim 18, wherein the dosage form of the pharmaceutical composition is an injection, tablet, suppository, ointment, gel, pill, tablet, granule, capsule or pharmaceutical preparation, suspension, or powder.

20. Use of the active site according to any one of claims 1 to 8, the Abelmoschus manihot extract according to any one of claims 9 to 13, the composition according to any one of claims 14 to 16, or the pharmaceutical composition according to any one of claims 17 to 19 in the preparation of a drug for nephropathy, preferably diabetic nephropathy or diabetic nephropathy or nephritis accompanied by renal fibrosis.

21. Use of the active site according to any one of claims 1 to 8, the Abelmoschus manihot extract according to any one of claims 9 to 13, the composition according to any one of claims 14 to 16, or the combination of drugs according to any one of claims 17 to 19 in the preparation of a drug for ophthalmological diseases, preferably for macular degeneration, visual fatigue or cataract, diabetic retinopathy, age-related macular degeneration, central exudative chorioretinitis, and macular degeneration due to severe myopia.

22. Use of an active site according to any one of claims 1 to 8, an Abelmoschus manihot extract according to any one of claims 9 to 13, a composition according to any one of claims 14 to 16, or a combination of drugs according to any one of claims 17 to 19 in the preparation of a drug for treating lupus erythematosus nephritis, or contrast-induced nephropathy, or pulmonary fibrosis, or heart failure, or for lowering uric acid.

23. A method for preparing the active site according to any one of claims 1 to 8 or the Abelmoschus maniihot extract according to any one of claims 9 to 13, comprising: (1) extracting Abelmoschus manii flowers or medicinal parts of Abelmoschus manii with ethanol to prepare an extract; (2) concentrating the extract and then extracting the extract to prepare an extract; (3) removing the solvent from the extract, and then eluting the extract with a macroporous resin to prepare a flavonoid active site or extract of Abelmoschus maniihot flowers; wherein preferably the extraction method is percolation.

24. The amount of ethanol used in step (1) is 10 to 25 times the amount of the medicinal site of the Abelmoschus mani hot flower or Abelmoschus mani hot, and the ethanol is a 60 to 95% ethanol solution; The extracting agent for the extract in step (2) is n-butanol, petroleum ether or ethyl acetate, the method of the extracting is continuous countercurrent extracting, the material liquid ratio of the extracting is 0.8-4:1, and the number of extracting stages of the extracting is 1-5; The type of the macroporous resin in step (3) is D101, HPD100 or AB-8; 24. The method of claim 23.

25. The preparation method according to claim 24, characterized in that step (2) further comprises subjecting the extraction liquid to activated carbon adsorption, alcohol precipitation or acid precipitation treatment prior to the extraction.

26. The preparation method according to claim 25, characterized in that the elution process of the macroporous resin is as follows: diameter-to-height ratio of the macroporous resin is 1:4-1:9; loading buffer concentration is 0.10-0.30 g raw material / mL; loading buffer volume is 4-12 BV; loading at a flow rate of 1-4 BV / h; removing impurities with 4-8 BV of pure water and 1-5 BV of 3-15% ethanol at a flow rate of 0.5-4 BV / h; eluting with 2-8 BV of 50-80% ethanol at a flow rate of 1-5 BV / h.

27. A method for preparing the active site according to any one of claims 1 to 8 or the Abelmoschus maniihot extract according to any one of claims 9 to 13, comprising: (1) extracting Abelmoschus manii flowers or medicinal parts of Abelmoschus manii with ethanol to prepare an extract; (2) adding a clarifying agent to the extract, subjecting it to a water bath treatment, and filtering to remove the supernatant; (3) Eluting the supernatant with a polyamide resin to prepare a flavonoid active part or extract of Abelmoschus manihot flower. wherein preferably the extraction method is percolation.

28. The amount of ethanol used in step (1) is 10 to 25 times the amount of the medicinal site of the Abelmoschus mani hot flower or Abelmoschus mani hot, and the ethanol is a 60 to 95% ethanol solution; In step (2), the water bath temperature is 50 to 70° C., and the water bath time is 30 to 90 minutes; In step (3), the diameter-to-height ratio of the resin is 1:4-1:9, the loading buffer concentration is 0.10-0.60 g starting material / mL, the loading buffer volume is 4-12 BV, and elution is performed using 4-8 BV of pure water and 4-8 BV of 60-95% ethanol; 28. The method of claim 27.

29. A method for preparing the active site according to any one of claims 1 to 8 or the Abelmoschus maniihot extract according to any one of claims 9 to 13, comprising: (1) extracting Abelmoschus manii flowers or medicinal parts of Abelmoschus manii with ethanol to prepare an extract; (2) adjusting the pH value of the extract to 2.0-3.0, refrigerating, filtering to remove precipitate, and adding water to dissolve; (3) extracting the solution to prepare an extract; (4) removing the solvent from the extract and then eluting it with a polyamide resin to prepare a flavonoid active part or extract of Abelmoschus maniihot flower; and preferably the extraction method is reflux.

30. The amount of ethanol used in step (1) is 10 to 25 times the amount of Abelmoschus manii flower or Abelmoschus manii active 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 extracting agent for the extract in step (3) is n-butanol, petroleum ether or ethyl acetate, the method of the extracting is continuous countercurrent extracting, the material liquid ratio of the extracting is 0.8-4:1, and the number of extracting stages of the extracting is 1-5; In step (4), the diameter-to-height ratio of the resin is 1:4-1:9, the loading buffer concentration is 0.10-0.60 g starting material / mL, the loading buffer volume is 4-12 BV, and elution is performed using 4-8 BV of pure water and 4-8 BV of 60-95% ethanol; 30. The method of claim 29.

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