CN103860638A - Preparation method of sophora alopecuroide flavonoid composition and new medical application - Google Patents
Preparation method of sophora alopecuroide flavonoid composition and new medical application Download PDFInfo
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Abstract
A sophora alopecuroide flavonoid composition is extracted and separated from dry mature seeds of (i) Sophora alopecuroide L. for the first time. The composition disclosed by the invention is proved to have the effects of inhibiting alpha-glucosaccharase activity and inhibiting lipase activity, and can be applied to prevention and treatment of diabetes and hyperlipidemia for the first time. Meanwhile, the invention relates to a medicament preparation of the medicament, which can be prepared into oral preparations, such as a hard capsule, a soft capsule, a tablet, granula, powder, a pill, a tea bag and a medicinal liquor.
Description
Technical field
The present invention relates to a kind of preparation method of extracting pharmaceutical composition from Herba Sophorae alopecuroidis and this pharmaceutical composition in the application of preparing in alpha-glucosidase inhibitor medicine and lipase inhibitor medicine, can be used for prevention and treatment diabetes and hyperlipidemia effect.The present invention also relates to the pharmaceutical preparation of this medicine, belong to medical technical field.
Background technology
Diabetes, hyperlipidemia are metabolic disease, and diabetes are often with disorders of lipid metabolism and hyperlipemia.At present, the sickness rate of metabolic disease is more and more higher, can relate to the chronic progressive external pathological changes of the tissues such as the heart, brain, kidney, lung, blood vessel, nerve, skin, eye, ear, foot, causes functional defect and exhaustion, and serious threat is to people's orthobiosis.Long-term hyperglycemia, very easily induced Diabetic cardio cerebrovascular affection of hyperlipidemia.Alpha-glucosidase inhibitor is by the activity of competitive inhibition alpha-glucosidase, can block disaccharidase and be hydrolyzed to monosaccharide, delay sugared absorption, make blood glucose steadily also maintain slowly certain level, control postprandial hyperglycemia disease, can also improve carbohydrate tolerance, prevention and treatment of obesity simultaneously, hypertriglyceridemia, is used for the treatment of the disease of answering carbohydrate disorder and cause.Lipase inhibitor is exactly the fat-reducing dietic medicinal being used for specially for lipase.Under the effect of lipase inhibitor, lipase will lose the capacity of decomposition of part, thereby nearly 1/3rd fat do not decompose by lipase, is excreted with food slag.It is external that lipase inhibitor is exactly that its part fat that people can be taken in just discharged before entering body blood, can control from fatty source like this accumulation of fat in human body.Moreover, the natural drug of these two kinds of metabolic diseases for the treatment of of the little determined curative effect of being also free from side effects at present.Diabetes and hyperlipidemia are commonly encountered diseases in recent years, the present invention is based on the drug regimen extracting in Herba Sophorae alopecuroidis and have Inhibiting α-glucosidase and suppress the dual function of lipase, said composition has not only can reduce blood glucose but also effect that can anti-curing hyperlipemia.
The present composition is from leguminous plant Herba Sophorae alopecuroidis
sophora alopecuroidesl. dry mature seed extracts to separate and obtains.Herba Sophorae alopecuroidis is important dimension medicine resource, and main product, in NORTHWEST CHINA area, contains the nutrient substance such as protein, saccharide, organic acid, flavonoid, pigment and alkaloid.At present the research of Herba Sophorae alopecuroidis is mainly concentrated on to Alkaloids From Sophora Alopecuroides L aspect, less to flavone research.The Herba Sophorae alopecuroidis gastric abscess that is used for the treatment of among the people, acid regurgitation, or join Herba Taraxaci, Rhizoma Zingiberis Recens etc. medicinal (" Chinese Herbal Medicine in Xinjiang handbook "), also can treat eczema, stubborn dermatitis; Among the people by radical cure treatment laryngalgia, cough, dysentery and eczema etc.; Yellow magnificent patent report (publication number: CN101982171A) bitter bean bisflavone there is antiviral and anti-tumor activity; Chen Hong patent of invention has reported that the compositions that (publication number: CN101480472B) is made up of Herba Sophorae alopecuroidis, Semen Cassiae, Fructus Crataegi, Radix Pseudostellariae, Rhizoma Gastrodiae has effect for reducing blood fat; A Jiaike Baeyer Chinese mugwort Sa patent of invention has reported that (CN1640429A) Herba Sophorae alopecuroidis seed or aerial parts n-butyl alcohol extract have hypolipidemic activity, n-butyl alcohol extract of the present invention is a kind of mixture, there is no clearly concrete effective substance, simultaneously, there is no clear and definite blood fat reducing mechanism, have significantly different from the present composition yet.
The present invention is the flavone composition extracting from Herba Sophorae alopecuroidis, also there is no bibliographical information about the preparation method of the flavone composition extracting from Herba Sophorae alopecuroidis and had Inhibiting α-glucosidase activity and the report of inhibition lipase active before the present invention completes.
Summary of the invention
Goal of the invention is to provide a kind of Herba Sophorae alopecuroidis flavone composition, for preventing and treat the effect of diabetes and hyperlipidemia.The present invention is studied the effective ingredient of Herba Sophorae alopecuroidis, has searched out active stronger, practical effective site, for prevention and the treatment of diabetes and hyperlipidemia.Further, the present inventor is by further research discovery, and the compositions being mainly made up of flavones ingredient is at Inhibiting α-glucosidase and suppress there is very strong activity aspect lipase.This natural pharmaceutical composition is that inventor's first passage experiment screening obtains, and has outstanding contribution and significant technological progress.
Outstanding contributions of the present invention are: enrichment flavone from dimension medicine resource Herba Sophorae alopecuroidis mature seed, obtain Herba Sophorae alopecuroidis flavone composition, and there is the effect for the treatment of diabetes and hyperlipidemia.A kind of new medicine resource and novel medical use are provided, and this is the most outstanding contribution of the present invention and the most significant novelty.
Two of the object of the invention has been to provide Herba Sophorae alopecuroidis flavone composition extraction separation method, medicinal part utilizes ethanol extraction, and extracting solution directly, by macroporous resin adsorption purification, is removed after the oil-soluble impuritieses such as pigment, extracting solution and eluent are through concentrated, and crystallize obtains micromolecule chromocor compound; Water-soluble flavone, alkaloid and with water-solubility impurity through polyamide adsorption and purification, remove the water-solubility impurities such as alkaloid, obtain macromole chromocor compound, merge two parts flavone, obtain the present composition, preparation method of the present invention is not used organic reagent, workable, is again one of innovative point of the present invention.
Another object of the present invention is to provide a kind of preparation method of the flavone composition extracting from Herba Sophorae alopecuroidis mature seed, it is characterized in that: the fragmentation of Herba Sophorae alopecuroidis seed, first doubly measure alcohol reflux 2-3 time of the 40-75% of medical material weight, each 2-3 hour with 4-10, filter, filtrate is by processed good macroporous resin column or polyamide column, then uses the ethanol elution 4-6 times column volume of 40-75%, collects effluent and eluent, reclaim ethanol, concentrated, centrifugal, precipitation is dried to obtain components I; Supernatant, by polyamide resin column, first washes with water, then uses the ethanol elution 4-6 times column volume of 40-75%, collects ethanol elution, concentrated, is dried to obtain compositionⅱ; Merge components I and compositionⅱ, obtain Herba Sophorae alopecuroidis flavone composition.
Preferred best preparation technology: Herba Sophorae alopecuroidis seed fragmentation, first with 65% alcohol reflux of 6 times of amount medical material weight 2-3 time, each 2-3 hour, filters, and filtrate is by processed good D101 macroporous resin column, use again 65% ethanol elution 4-6 times column volume, collect effluent and eluent, reclaim ethanol, concentrated, centrifugal, precipitation is dried to obtain components I; Supernatant, by polyamide resin column, first washes with water, then uses 4 times of column volumes of ethanol elution of 70%, collects ethanol elution, concentrated, is dried to obtain compositionⅱ; Merge components I and compositionⅱ, obtain Herba Sophorae alopecuroidis flavone composition.
Another outstanding contributions of the present invention are, a kind of pharmaceutical composition that has Inhibiting α-glucosidase activity and suppress lipase active is provided, and it is characterized in that: in compositions, general flavone content is not less than 80%.
For realizing above technical scheme, Herba Sophorae alopecuroidis flavone composition general flavone content is measured by the following method:
The preparation precision of reference substance solution takes control substance of Rutin 10mg, puts in 100ml measuring bottle, with dissolve with methanol, and is diluted to scale, shakes up, and precision measures 5ml and puts in 10ml measuring bottle, is diluted to scale with methanol, shakes up, and to obtain final product.
Herba Sophorae alopecuroidis flavone composition powder is got in the preparation of need testing solution, and accurately weighed 50mg, to 50ml volumetric flask, adds methanol appropriate, supersound process 20min, takes out, and places room temperature, adds methanol constant volume to scale, shake up, get in 2ml to 25ml volumetric flask, add methanol constant volume to scale and get final product.
Algoscopy is got respectively reference substance solution, need testing solution, retinue blank.According to spectrophotography (annex V B of " Chinese Pharmacopoeia " version in 2010), measure respectively trap at wavelength 340nm place, calculate.
Herba Sophorae alopecuroidis flavone composition (abbreviation compositions) has the active of Inhibiting α-glucosidase and suppresses lipase active, is confirmed by following pharmacodynamic experiment.
Experimental raw: the compositions of using in this pharmacodynamics, for the inventor provides, is prepared from according to example 1.
The orlistat capsule of using in this pharmacodynamics test is that (lot number is Haizheng Medicine Stock Co., Ltd., Zhejiang Prov's production: 20130302); Acarbose is that Bayer HealthCare Co produces (lot number: 20130905).
The alpha-glucosidase that this pharmacodynamics test is used is that sigma company produces (G5003-100UN); Mucosa enzyme self-control in rat intestine; Lipase is that Beijing CTA New Century Biotechnology Co. Ltd. produces.
1, the inhibitory action of Herba Sophorae alopecuroidis flavone composition to alpha-glucosidase
1.1 standard reaction system 67mmol/L pH are 6.8 kaliumphosphate buffer 150uL, 1mg/mL glutathione solution 50 μ L, 2.5U/mL alpha-glucosaccharase enzymatic solution 100 μ L, 37 DEG C of insulation 10min, 20mmol/L PNPG solution 100ul, adds 0.2mol/L Na after 37 DEG C of reaction 20min
2cO
3solution 400uL cessation reaction, measures 400nm place absorbance value.The suppression ratio computing formula of sample to enzymatic activity:
The acarbose that suppression ratio (%)=(A contrast-A sample)/A contrast × 100% 1.2 acarbose is got after grinding the impact of alpha-glucosidase activity is mixed with the original solution of 50mg/ml, and is diluted to 10.0,5.0 with this, 2.5,1.0,0.2,0.1,0.05mg/ml variable concentrations.100ul acarbose is joined in enzyme reaction system, be first incubated 10min with enzyme at 37 DEG C, then add substrate reactions 20min, use Na
2cO
3solution cessation reaction, measures 400nm place light absorption value.
1.3 Herba Sophorae alopecuroidis flavone compositions affect compositions dmso solution to alpha-glucosidase activity, are then diluted to 5.0,2.5 with buffer, and 1.0,0.5,0.25,0.1,0.05 mg/ml, dimethyl sulfoxide is whole content ﹤ 1% in reaction system.100ul compositions is joined in reaction system, measure the inhibitory action of compositions to alpha-glucosidase.Experimental result is in table 1.
The inhibitory action of table 1 Herba Sophorae alopecuroidis flavone composition to alpha-glucosidase
Result shows, acarbose has good dose-effect relationship to the inhibitory action of alpha-glucosidase activity.Compositions is 5.0 in concentration, 2.5,1.0,0.5,0.25,0.1, under 0.05 mg/ml condition, the inhibitory action of alpha-glucosidase is existed to good dose-effect relationship, and compositions is the more than 2 times of positive drug acarbose to the inhibitory action of alpha-glucosidase activity, and alpha-glucosidase is shown to very strong inhibitory action.
2, the inhibitory action of Herba Sophorae alopecuroidis flavone composition to mucosa enzyme in rat intestine
The extraction of mucosa enzyme in 2.1 rat intestines
Get 1 of normal rat, after execution, take out immediately small intestinal, clean content, add the 0.1mol/L of 4 DEG C of pre-coolings by mass volume ratio 1:3, pH=6.8 PBS, homogenate, 4 DEG C, 8000 rpm, centrifugal 20 minutes, get supernatant and be distributed in EP pipe ,-20 DEG C are freezing for subsequent use.
The inhibitory action of 2.2 Herba Sophorae alopecuroidis flavone compositions to mucosa enzyme in rat intestine
2.2.1 standard reaction system 67mmol/L pH is 6.8 kaliumphosphate buffer 150uL, 1mg/mL glutathione solution 50 μ L, mucosa enzymatic solution 100 μ L in rat intestine, 37 DEG C of insulation 10min, 20mmol/L PNPG solution 100ul, adds 0.2mol/L Na after 37 DEG C of reaction 20min
2cO
3solution 400uL cessation reaction, measures 400nm place absorbance value.The suppression ratio computing formula of sample to enzymatic activity:
The acarbose that suppression ratio (%)=(A contrast-A sample)/A contrast × 100% 2.2.2 acarbose is got after grinding the impact of mucosa enzymatic activity in rat intestine is mixed with the original solution of 50mg/ml, and is diluted to 10.0,5.0 with this, 2.5,1.0,0.2,0.1,0.05mg/ml variable concentrations.100ul acarbose is joined in enzyme reaction system, be first incubated 10min with enzyme at 37 DEG C, then add substrate reactions 20min, use Na
2cO
3solution cessation reaction, measures 400nm place absorbance value.
2.2.3 Herba Sophorae alopecuroidis flavone composition on mucosa enzymatic activity in rat intestine affect compositions dmso solution, be then diluted to 5.0,2.5 with buffer, 1.0,0.5,0.25,0.1,0.05 mg/ml, dimethyl sulfoxide is whole content ﹤ 1% in reaction system.100ul compositions is joined in reaction system, measure the inhibitory action of compositions to mucosa enzyme in rat intestine.Experimental result is in table 2.
The inhibitory action of table 2 Herba Sophorae alopecuroidis flavone composition to mucosa enzyme in rat intestine
Result shows, acarbose has good dose-effect relationship to the inhibitory action of mucosa enzymatic activity in rat intestine.Compositions is 5.0 in concentration, 2.5,1.0,0.5,0.25,0.1, under 0.05mg/ml condition, there is good dose-effect relationship in the inhibitory action to mucosa enzyme in rat intestine, compositions is the more than 2 times of positive drug acarbose to the inhibitory action of mucosa enzymatic activity in rat intestine, and mucosa enzyme in rat intestine is shown to very strong inhibitory action.
3, the inhibitory action of Herba Sophorae alopecuroidis flavone composition to lipase
The mensuration of 3.1 lipase inhibiting activities
In 100mL conical flask, add 5mL 0.025mol/L phosphate buffer (pH=7.4) and 4mL polyvinyl alcohol glycerol trioleate substrate emulsion (0.229g/mL), be placed in 37 DEG C of water bath heat preservation 10min, then add 1mL enzyme liquid (2mg/mL), react again afterwards 15min, add 95% ethanol 15mL and stop enzyme reaction.Add 2, phenolphthalein, be titrated to blush with 0.025mol/LNaOH standard solution.The enzyme liquid of blank experiment adds after reaction terminating.
3.2 positive control drug orlistats are to lipase inhibitory action
In 100mL conical flask, add 5mL 0.025mol/L phosphate buffer (pH=7.4) and 4mL polyvinyl alcohol glycerol trioleate substrate emulsion (0.229g/mL), adding respectively concentration is 5.0,2.5,1.0,0.5, the positive control drug 1mL of 0.25,0.1,0.05mg/ml, be placed in 37 DEG C of water bath heat preservation 10min, then add 1mL enzyme liquid (2mg/mL), react again afterwards 15min, add 95% ethanol 15mL and stop enzyme reaction.Add 2, phenolphthalein, be titrated to blush with 0.025mol/LNaOH standard solution.
3.3 Herba Sophorae alopecuroidis flavone compositions are to lipase inhibitory action
In 100mL conical flask, add 5mL 0.025mol/L phosphate buffer (pH=7.4) and 4mL polyvinyl alcohol glycerol trioleate substrate emulsion (0.229g/mL), adding respectively concentration is 5.0,2.5,1.0,0.5, the Herba Sophorae alopecuroidis flavone composition 1mL of 0.25,0.1,0.05mg/ml, be placed in 37 DEG C of water bath heat preservation 10min, then add 1mL enzyme liquid (2mg/mL), react again afterwards 15min, add 95% ethanol 15mL and stop enzyme reaction.Add 2, phenolphthalein, be titrated to blush with 0.025mol/LNaOH standard solution.
Suppression ratio=
× 100%
The inhibitory action of table 3 Herba Sophorae alopecuroidis flavone composition to lipase
Result shows, orlistat has good dose-effect relationship to the inhibitory action of lipase.Compositions is 5.0,2.5,1.0,0.5,0.25,0.1,0.05 in concentration, under condition, the inhibitory action of lipase is existed to good dose-effect relationship, compositions to the inhibitory action of lipase active compared with positive drug orlistat a little less than.
Embodiment
The present invention executes example by following experiment to be achieved (confirmation), but is not limited only to this example.
the preparation method of example 1 Herba Sophorae alopecuroidis flavone composition
Herba Sophorae alopecuroidis seed 10kg fragmentation, first with 65% alcohol reflux of 6 times of amount medical material weight 3 times, each 2 hours, filter, filtrate is by processed good D101 macroporous resin column, use again 5 times of column volumes of ethanol elution of 65%, collect effluent and eluent, reclaim ethanol, concentrated, centrifugal, precipitation is dried to obtain components I; Supernatant, by polyamide resin column, first washes 5 times of column volumes with water, then uses 4 times of column volumes of ethanol elution of 70%, collects ethanol elution, concentrated, is dried to obtain compositionⅱ; Merge components I and compositionⅱ, obtain Herba Sophorae alopecuroidis flavone composition 295g, general flavone content is 86.2%.
the preparation method of example 2 Herba Sophorae alopecuroidis flavone compositions
Herba Sophorae alopecuroidis seed 10kg fragmentation, first with 60% alcohol reflux of 8 times of amount medical material weight 2 times, each 2 hours, filter, filtrate is by processed good D101 macroporous resin column, use again 5 times of column volumes of ethanol elution of 60%, collect effluent and eluent, reclaim ethanol, concentrated, centrifugal, precipitation is dried to obtain components I; Supernatant, by polyamide resin column, first washes 4 times of column volumes with water, then uses 4 times of column volumes of ethanol elution of 80%, collects ethanol elution, concentrated, is dried to obtain compositionⅱ; Merge components I and compositionⅱ, obtain Herba Sophorae alopecuroidis flavone composition 268g, general flavone content is 83.7%.
embodiment 3(capsule)
Composition material 100g, medical starch is appropriate, and mix homogeneously is granulated, and dry, granulate, packs capsule into No. 1, makes 1000, to obtain final product.Each 3, every day 2 times.
embodiment 4(tablet)
Composition material 100g, medical starch is appropriate, and dextrin is appropriate, and mix homogeneously is granulated, dry, and granulate is dry, adds lubricant appropriate, and tabletting, makes 1000, to obtain final product.Each 3, every day 2 times.
Claims (7)
1. the Herba Sophorae alopecuroidis flavone composition extracting from Herba Sophorae alopecuroidis mature seed has the application in the inhibiting medicine of alpha-glucosidase in preparation.
2. the Herba Sophorae alopecuroidis flavone composition extracting from Herba Sophorae alopecuroidis mature seed has the application in the inhibiting medicine of lipase in preparation.
3. according to the preparation method of the Herba Sophorae alopecuroidis flavone composition described in claim 1 and 2, it is characterized in that: the fragmentation of Herba Sophorae alopecuroidis seed, first doubly measure alcohol reflux 2-3 time of the 40-75% of medical material weight, each 2-3 hour with 4-10, filter, filtrate is by processed good macroporous resin column or polyamide column, then uses the ethanol elution 4-6 times column volume of 40-75%, collects effluent and eluent, reclaim ethanol, concentrated, centrifugal, precipitation is dried to obtain components I; Supernatant, by polyamide resin column, first washes with water, then uses the ethanol elution 4-6 times column volume of 40-75%, collects ethanol elution, concentrated, is dried to obtain compositionⅱ; Merge components I and compositionⅱ, obtain Herba Sophorae alopecuroidis flavone composition.
4. Herba Sophorae alopecuroidis flavone composition preparation method according to claim 3, it is characterized in that: Herba Sophorae alopecuroidis seed 10kg fragmentation, first with 65% alcohol reflux of 6 times of amount medical material weight 3 times, each 2 hours, filter, filtrate is by processed good D101 macroporous resin column, then uses 5 times of column volumes of ethanol elution of 65%, collects effluent and eluent, reclaim ethanol, concentrated, centrifugal, precipitation is dried to obtain components I; Supernatant, by polyamide resin column, first washes 5 times of column volumes with water, then uses 4 times of column volumes of ethanol elution of 70%, collects ethanol elution, concentrated, is dried to obtain compositionⅱ; Merge components I and compositionⅱ, obtain Herba Sophorae alopecuroidis flavone composition.
5. according to the Herba Sophorae alopecuroidis flavone composition described in any one in claim 1-4, it is characterized in that: the content of total flavones is no less than 80%.
6. an oral drug preparation, is characterized in that: the Herba Sophorae alopecuroidis flavone composition that contains the treatment effective dose described in any one in claim 1-5 and one or more pharmaceutically acceptable excipient.
7. pharmaceutical preparation according to claim 6, is characterized in that: described oral formulations is selected from the one in the preparations such as tablet, capsule, granule, pill, suspensoid, drop pill, oral liquid.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105832718A (en) * | 2016-04-26 | 2016-08-10 | 宁夏医科大学 | Application of sophoraflavanone G in preparation of medicine for preventing and treating diabetic mellitus |
| CN105832814A (en) * | 2016-04-26 | 2016-08-10 | 宁夏医科大学 | General flavone with effect of treating diabetic mellitus, as well as preparation method and application thereof |
| CN106243120A (en) * | 2016-06-29 | 2016-12-21 | 塔里木大学 | The preparation of Herba Sophorae alopecuroidis flavone extract and the application in cosmetics thereof |
| CN111202765A (en) * | 2019-11-19 | 2020-05-29 | 宁夏医科大学 | Anti-depression total flavone extract and preparation method and application thereof |
| CN111249271A (en) * | 2020-03-19 | 2020-06-09 | 中南民族大学 | Medicament for treating diabetes and preparation method and application thereof |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105832718A (en) * | 2016-04-26 | 2016-08-10 | 宁夏医科大学 | Application of sophoraflavanone G in preparation of medicine for preventing and treating diabetic mellitus |
| CN105832814A (en) * | 2016-04-26 | 2016-08-10 | 宁夏医科大学 | General flavone with effect of treating diabetic mellitus, as well as preparation method and application thereof |
| CN105832718B (en) * | 2016-04-26 | 2018-08-21 | 宁夏医科大学 | Flavones Sophoraflavanone G are preparing the application in preventing diabetes medicament |
| CN106243120A (en) * | 2016-06-29 | 2016-12-21 | 塔里木大学 | The preparation of Herba Sophorae alopecuroidis flavone extract and the application in cosmetics thereof |
| CN106243120B (en) * | 2016-06-29 | 2019-04-19 | 塔里木大学 | The preparation of Sophora alopecuroide flavones Stilbene class extract and its application in cosmetics |
| CN111202765A (en) * | 2019-11-19 | 2020-05-29 | 宁夏医科大学 | Anti-depression total flavone extract and preparation method and application thereof |
| CN111249271A (en) * | 2020-03-19 | 2020-06-09 | 中南民族大学 | Medicament for treating diabetes and preparation method and application thereof |
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| CN103860638B (en) | 2017-02-15 |
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