CN102091082A - Ginsenoside Rb3 composition and application thereof in preparing medicament for treating diabetes - Google Patents
Ginsenoside Rb3 composition and application thereof in preparing medicament for treating diabetes Download PDFInfo
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- CN102091082A CN102091082A CN 201110005851 CN201110005851A CN102091082A CN 102091082 A CN102091082 A CN 102091082A CN 201110005851 CN201110005851 CN 201110005851 CN 201110005851 A CN201110005851 A CN 201110005851A CN 102091082 A CN102091082 A CN 102091082A
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- TXEWRVNOAJOINC-UHFFFAOYSA-N ginsenoside Rb2 Natural products CC(=CCCC(OC1OC(COC2OCC(O)C(O)C2O)C(O)C(O)C1O)C3CCC4(C)C3C(O)CC5C6(C)CCC(OC7OC(CO)C(O)C(O)C7OC8OC(CO)C(O)C(O)C8O)C(C)(C)C6CCC45C)C TXEWRVNOAJOINC-UHFFFAOYSA-N 0.000 title claims abstract description 71
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Abstract
The invention relates to a ginsenoside Rb3 composition and an application thereof in preparing a medicament for treating diabetes. The ginsenoside Rb3 composition is prepared from ginsenoside Rb3 as a main component and 1-5 of a carrier, an excipient, a sweetening agent, an adhesive, a flavoring agent, a preservative, a dispersing agent, a surfactant and a colorant as adjuvants, and the medicament is prepared into tablets, pills, granules, capsules and powder. The method for extracting the ginsenoside Rb3 comprises the following steps: dissolving crude extracts of stems and leaves of American ginseng, ginseng or pseudo-ginseng in water, then separating by NKA-12 macroporous resin, and performing gradient elution by ethanol water to obtain panaxatriol saponin and panaxadiol saponin. Repeated chromatography is preformed on the panaxadiol saponin by silicagel columns to obtain pure ginsenoside Rb3 with the purity of 98%. Proven by animal experiments, the ginsenoside Rb3 has notable effects of symptom improvement and treatment on diabetes and adiposis.
Description
Technical field
The invention belongs to pharmaceutical product field category, relate to the extract that derives from the plant Radix Ginseng as the application of medicine to the treatment diabetes.
Background technology
Relevant Radix Ginseng The Chemical Constituents is the history in existing over one hundred year so far.The chemical constituent more complicated of Radix Ginseng comprises saponin, saccharide, protein, polypeptide, aminoacid, organic acid, vitamin, volatilization wet goods composition.Wherein the ginsenoside is the main active of Radix Ginseng.Many Chinese scholars have been studied the effect of Radix Ginseng aspect anti-diabetic and fat-reducing, have wherein both comprised Radix Ginseng, Radix Ginseng extract, panoxadiol's group and triol saponins, also comprise the monomer ginsenoside.The effect of ginsenoside Rb3 aspect anti-diabetic and antiobesity action in vivo do not have report as yet.
Summary of the invention
The object of the present invention is to provide a kind of Panaxsaponin composition, it is to improving glucose tolerance, improve kidney of diabetic patients, heart, nervous system and retinal complication, improving the effective product of diabetics hyperinsulinemia.
Pharmaceutical composition among the present invention wherein contains the pure product 1%-20% of ginsenoside Rb3, also contains other ginsenosides, polysaccharide, protein, aminoacid, trace element etc.It will be appreciated by those skilled in the art that the effect of pharmaceutical composition of the present invention can be kept or strengthen to the existence of these materials.
Ginsenoside Rb3 method of purification: Radix Panacis Quinquefolii (perhaps Radix Ginseng, Radix Notoginseng) stem and leaf crude extract, separate with the NKA-12 macroporous resin water-soluble back, ethanol water gradient elution, obtain panaxatriol's saponins and panoxadiol's saponins, panoxadiol's saponins usefulness silicagel column (chloroform-methanol-water, n-butyl alcohol-ethyl acetate-water system) chromatography repeatedly obtains the pure product of ginsenoside Rb3 (purity 98%).
In addition, what should particularly point out is, can be as required in the drug regimen of invention, add one or more natural or synthetic other and have the composition of collaborative or assosting effect with this active substance, the natural or synthetic auxiliary element that these may be added into be well known by persons skilled in the art with can imagine.Said compositions can be mixed with peroral administration dosage form, as tablet, pill, granule, capsule, powder etc.
In order to prepare the tablet that is suitable for oral administration, pill, granule, capsule etc., can use sucrose, galactose, corn starch, gelatin, microcrystalline cellulose, Pulvis Talci etc. as carrier or excipient.In these peroral administration dosage forms, can also contain other suitable additive, such as disintegrating agent, lubricant, filler, adhesive, correctives, antiseptic, dispersant, surfactant, coloring agent etc.
The most preferred route of administration of pharmaceutical composition of the present invention is the dosage form of various oral administrations, for example tablet, granule, capsule or pill, and dosage every day of these oral administration dosage forms generally contains this active substance of 10～1000mg.Following experiment has further proved the pharmacological action of medicine of the present invention.
Ginsenoside Rb3 shows zoopery: to the influence of alloxan type diabetic mice fasting glucose, oral glucose tolerance, body weight, food ration and amount of drinking water, have certain blood sugar lowering and the function of improving glucose tolerance.Simultaneously ginsenoside Rb3 shows the influence experiment of the inductive obesity mice of high fat diet: reduce in the inductive obesity mice serum of high fat diet that T-CHOL contains and the uterus around fat weight, and improve the inductive obesity mice oral glucose of high fat diet tolerance, be that the prescription of primary raw material has the effect because of the fat insulin resistant that produces of certain antiobesity action and prevention or treatment with ginsenoside Rb3.
Description of drawings
Fig. 1: ginsenoside Rb3 is to the figure that influences of alloxan type diabetic mice fasting glucose.
Fig. 2: ginsenoside Rb3 is to the figure that influences of alloxan type diabetic mice carbohydrate tolerance.
Fig. 3: ginsenoside Rb3 is to the figure that influences of alloxan type diabetic mice food ration.
Fig. 4: ginsenoside Rb3 is to the figure that influences of alloxan type diabetic mice amount of drinking water.
Fig. 5: ginsenoside Rb3 is to the figure that influences of total cholesterol level in the high fat diet mice serum.
Fig. 6: ginsenoside Rb3 is to the figure that influences of fat weight around the high fat diet Mouse Uterus.
Fig. 7: ginsenoside Rb3 is to the figure that influences of high fat diet mice carbohydrate tolerance.
The specific embodiment
One. ginsenoside Rb3 is to zoopery:
Experimental example 1 ginsenoside Rb3 is to the hypoglycemic activity of the inductive diabetic mice of alloxan:
1. experiment material, animal, instrument and reagent
1.1 experiment material
Radix Panacis Quinquefolii (perhaps Radix Ginseng, Radix Notoginseng) stem and leaf crude extract, separate with the NKA-12 macroporous resin water-soluble back, ethanol water gradient elution, obtain panaxatriol's saponins and panoxadiol's saponins, panoxadiol's saponins usefulness silicagel column (chloroform-methanol-water, n-butyl alcohol-ethyl acetate-water system) chromatography repeatedly obtains the pure product of ginsenoside Rb3 (purity 98%).
Animal is a Kunming mouse, and body weight 20-22g is female.Available from preclinical medicine institute of Jilin University Experimental Animal Center.
1.2 instrument
Doubly easy type blood glucose meter of steady person of outstanding talent and blood sugar test paper, Johnson ﹠ Johnson (China) medical apparatus and instruments company limited; The BS-400 automatic clinical chemistry analyzer, Mairui Biological Medical Electronic Co., Ltd., Shenzhen; BP211D type electronic balance (100,000/), German Sartorius company.
1.3 medicine and reagent
Ginsenoside Rb3 is made by oneself by this experimental group as described in 1.1.
Alloxan, Sigma company; Glucose, citric acid, sodium citrate, the Beijing Chemical Plant; Chloroform, methanol are analytical pure, the Beijing Chemical Plant; Normal feedstuff, preclinical medicine institute of Jilin University Experimental Animal Center.
2. test method
2.1 influence to alloxan type blood glucose in diabetic mice
Get 60 of healthy mices, get 8 at random as blank, it is normal group, the equal lumbar injection alloxan of all the other mices 200mg/kg, after 6 days, the mice of modeling success is divided into four groups (8 every group) at random: model control group, ginsenoside Rb3 5mg/kg, 15mg/kg, three dosage treatment groups of 25mg/kg.The grouping back is gastric infusion respectively, and every morning, 8:30-9:30 was administered once, successive administration 14 days.At the 0th day, the 7th day, survey in the 14th day is 12h blood glucose on an empty stomach respectively.
2.2 influence to alloxan type diabetic mice carbohydrate tolerance
Get 60 of healthy mices, get 8 at random as blank, it is normal group, the equal lumbar injection alloxan of all the other mices 200mg/kg, after 6 days, the mice of modeling success is divided into four groups (8 every group) at random: model control group, ginsenoside Rb3 5mg/kg, 15mg/kg, three dosage treatment groups of 25mg/kg.The grouping back is gastric infusion respectively, and every morning, 8:30-9:30 was administered once, successive administration 14 days.The 14th day, to irritate stomach and give mice glucose 2g/kg on an empty stomach, docking is got blood system and is not measured 0min, 30min, 60min, 120min blood glucose.
2.3 influence to alloxan type diabetic mice body weight
Get 60 of healthy mices, get 8 at random as blank, it is normal group, the equal lumbar injection alloxan of all the other mices 200mg/kg, after 6 days, the mice of modeling success is divided into four groups (8 every group) at random: model control group, ginsenoside Rb3 5mg/kg, 15mg/kg, three dosage treatment groups of 25mg/kg.The grouping back is gastric infusion respectively, and every morning, 8:30-9:30 was administered once, successive administration 14 days.Record mice body weight before administration every day.
2.4 influence to alloxan type diabetic mice food ration
Get 60 of healthy mices, get 8 at random as blank, it is normal group, the equal lumbar injection alloxan of all the other mices 200mg/kg, after 6 days, the mice of modeling success is divided into four groups (8 every group) at random: model control group, ginsenoside Rb3 5mg/kg, 15mg/kg, three dosage treatment groups of 25mg/kg.The grouping back is gastric infusion respectively, and every morning, 8:30-9:30 was administered once, successive administration 14 days.Every morning claims Mus grain, calculates food ration.
2.5 influence to alloxan type diabetic mice amount of drinking water
Get 60 of healthy mices, get 8 at random as blank, it is normal group, the equal lumbar injection alloxan of all the other mices 200mg/kg, after 6 days, the mice of modeling success is divided into four groups (8 every group) at random: model control group, ginsenoside Rb3 5mg/kg, 15mg/kg, three dosage treatment groups of 25mg/kg.The grouping back is gastric infusion respectively, and every morning, 8:30-9:30 was administered once, successive administration 14 days.Every morning is write down amount of drinking water.
3. experimental result
3.1 influence to alloxan type diabetic mice fasting blood glucose level
Measured mouse blood sugar respectively at the 0th day, the 7th day, the 14th day.The 0th day, ginsenoside Rb3 treatment group 5mg/kg, 15mg/kg, the blood sugar level of 25mg/kg and model group is all apparently higher than normal group.The 14th day, 15mg/kg group mouse blood sugar (6.58mmol/L) was compared with model group (10.40mmol/L) with 25mg/kg group mouse blood sugar (6.50mmol/L) and is reduced by 36.7% and 37.5% respectively, and the blood glucose of 25mg/kg group mice has recovered normal level.(seeing accompanying drawing 1)
3.2 influence to alloxan type diabetic mice oral glucose tolerance
Through treatment in 14 days, the carbohydrate tolerance of 15mg/kg group mice was compared obvious improvement with model group, and AUC reduces by 34.47% (P＜0.05).During 120min, the blood glucose of 15mg/kg group mice has recovered normal level.Yet the AUC of three dosage groups is still apparently higher than normal group.(seeing accompanying drawing 2)
3.3 influence to alloxan type diabetic mice body weight
Ginsenoside Rb3 is to the not significantly influence of alloxan type diabetic mice body weight.
3.4 influence to alloxan type diabetic mice food ration
Compare with model group, after the ginsenoside Rb3 treatment, the 15mg/kg group significantly reduces (P＜0.01) with the food ration of 25mg/kg group mice.(seeing accompanying drawing 3)
3.5 influence to alloxan type diabetic mice amount of drinking water
Through 5mg/kg, 15mg/kg, the treatment of three dosage ginsenosides of 25mg/kg Rb3, the amount of drinking water of diabetic mice is compared with model group all and is significantly reduced.(seeing accompanying drawing 4)
4. discuss
Ginsenoside Rb3 can reduce alloxan type blood glucose in diabetic mice, improves mice oral glucose tolerance, reduces diabetic mice food ration and amount of drinking water, yet ginsenoside Rb3 is to the not significantly influence of diabetic mice body weight.To sum up, ginsenoside Rb3 has certain blood sugar lowering and the function of improving glucose tolerance.
Experimental example 2 ginsenoside Rb3 are to the influence of the inductive obesity mice of high fat diet
1 experiment material, instrument and reagent
1.1 material
Ginsenoside Rb3 is prepared by this experimental group as described in the example 1 1.1, and Rb3 content is 20%.
1.2 instrument and reagent
The T-CHOL test kit is available from the middle north control bio tech ltd of giving birth to; The BS-400 automatic clinical chemistry analyzer, Mairui Biological Medical Electronic Co., Ltd., Shenzhen; BP211D type electronic balance (100,000/), German Sartorius company; Doubly easy type blood glucose meter of steady person of outstanding talent and blood sugar test paper, Johnson ﹠ Johnson (China) medical apparatus and instruments company limited.
1.3 laboratory animal
Animal is a Kunming mouse, and is female, and available from preclinical medicine institute of Jilin University Experimental Animal Center, body weight is at 19-22g, Mus 3 weeks of age.
1.4 animal feed
Conventional feed is Jilin University's Experimental Animal Center preparation, and total amount of heat is 5.23J/g.The self-control of high lipid food laboratory, total amount of heat is 14.6J/g.Feedstuff all is kept in the ultra cold storage freezer-30 ℃ and is in store for.
2. test method
2.1 the test of total cholesterol level in the mice serum
Get 24 of healthy mices, sub-cage rearing is in the receptacle of room temperature, and light and shade 12h circulation adapts to new environment of 1 week before the experiment.During this period, the mice normal diet of feeding, animal free choice feeding feedstuff and freely drinking water shaked down for 1 week.Get 8 at random as blank, i.e. normal group, the conventional feed of feeding; All the other mices are divided into two groups at random: model control group, the high lipid food of feeding; Ginsenoside Rb3 prevention group is fed and is added the high lipid food of 0.02% ginsenoside Rb3.Raised for 8 weeks.After 8 weeks, mice is plucked eyeball get blood, 2500r/min is centrifugal, and 10min gets supernatant, makes mice serum, adopts test kit to measure T-CHOL in the serum (TC) respectively by automatic clinical chemistry analyzer.
2.2 the weighing of fat around the Mouse Uterus
Get 24 of healthy mices, sub-cage rearing is in the receptacle of room temperature, and light and shade 12h circulation adapts to new environment of 1 week before the experiment.During this period, the mice normal diet of feeding, animal free choice feeding feedstuff and freely drinking water shaked down for 1 week.Get 8 at random as blank, i.e. normal group, the conventional feed of feeding; All the other mices are divided into two groups at random: model control group, the high lipid food of feeding; Ginsenoside Rb3 prevention group is fed and is added the high lipid food of 0.02% ginsenoside Rb3.Raised for 8 weeks.After 8 weeks, after mice plucked eyeball and get blood, the cervical vertebra dislocation was put to death, and takes out fat around the uterus, weighing.
2.3 the mensuration of mice carbohydrate tolerance
Get 24 of healthy mices, sub-cage rearing is in the receptacle of room temperature, and light and shade 12h circulation adapts to new environment of 1 week before the experiment.During this period, the mice normal diet of feeding, animal free choice feeding feedstuff and freely drinking water shaked down for 1 week.Get 8 at random as blank, i.e. normal group, the conventional feed of feeding; All the other mices are divided into two groups at random: model control group, the high lipid food of feeding; Ginsenoside Rb3 prevention group is fed and is added the high lipid food of 0.02% ginsenoside Rb3.Raised for 8 weeks.After 8 weeks, irritate stomach and give mice glucose 2g/kg on an empty stomach, docking is got blood system and is not measured 0min, 30min, 60min, 120min blood glucose.
2.4 statistical disposition
Data are represented with mean+SD.Carry out statistical analysis with SPSS software, use one factor analysis of variance to come analytical data, P＜0.05 is judged to be significant difference on the statistics, and it is extremely remarkable that P＜0.01 is judged to be on the statistics difference.
3. experimental result
3.1 influence to total cholesterol level in the inductive obesity mice serum of high fat diet
Feed after 8 weeks, total cholesterol level is apparently higher than normal group in the model group mice serum.Compare with model group, the TC in the ginsenoside Rb3 prevention group mice serum has reduced by 27.42%, has significant difference (P＜0.05).(seeing accompanying drawing 5)
3.2 influence to fat weight around the inductive obesity mice of the high fat diet uterus
Feed after 8 weeks, the model group Mouse Uterus is obvious height of fat weight and normal group on every side.Compare with model group, fat weight obviously reduces (P＜0.05) around the ginsenoside Rb3 prevention group mice Mouse Uterus.(seeing accompanying drawing 6)
3.3 influence to the inductive obesity mice carbohydrate tolerance of high fat diet
Feed after 8 weeks, ginsenoside Rb3 has obviously improved high fat diet mice carbohydrate tolerance, compares with model group, and AUC reduces by 24.43% (P＜0.05).In addition, during 30min, the blood glucose of ginsenoside Rb3 prevention group mice is compared significant difference (P＜0.05) with model group.(seeing accompanying drawing 7)
4. discuss
Based on the prescription of ginsenoside Rb3 can reduce in the inductive obesity mice serum of high fat diet that T-CHOL contains and the uterus around fat weight, and improve the inductive obesity mice oral glucose of high fat diet tolerance.To sum up, be that the prescription of primary raw material has the effect because of the fat insulin resistant that produces of certain antiobesity action and prevention or treatment with ginsenoside Rb3.
Two. the preparation of medicine
Embodiment 1: the extraction of ginsenoside Rb3
Radix Panacis Quinquefolii (Radix Ginseng, Radix Notoginseng) stem and leaf crude extract 200g, separate with the NKA-12 macroporous resin water-soluble back, and 30%, 40%, 50% ethanol gradient elution obtains panaxatriol's saponins and panoxadiol's saponins.Resulting panoxadiol's saponins with silicagel column repeatedly chromatography (chloroform-methanol-water, n-butyl alcohol-ethyl acetate-water system) obtain the pure product 10.5g of ginsenoside Rb3.
Embodiment 2: the preparation of blood sugar lowering tablet
Get the pure product 2g of embodiment 1 ginsenoside Rb3, cyclodextrin 20g, dry starch is an amount of, and carboxymethyl cellulose is an amount of, and Pulvis Talci is an amount of, and sucrose is an amount of.Sucrose and Pulvis Talci are used for sugar coating, mixing, and pelletizing press sheet makes the heavy 0.50g of tablet, contains ginsenoside Rb3 1%-5% (g/g).Other requirements meet the relevant regulations of 2010 editions relevant tablets of the Pharmacopoeia of the People's Republic of China.
Embodiment 3: improve the capsular preparation of carbohydrate tolerance
Get embodiment 1 active substance 1g, corn starch 20g, cyclodextrin is an amount of, and mixing incapsulates, and every contains ginsenoside Rb3 1-10% (g/g).Other requirements meet the relevant regulations of 2010 editions relevant capsules of the Pharmacopoeia of the People's Republic of China.
Embodiment 4: the preparation of blood-sugar reducing tea
Get embodiment 1 ginsenoside Rb3 10g, dextrin 100g, soluble starch 100g, Icing Sugar is an amount of, mixing, the tea making piece divides packing, loading amount 5.0g/ bag.Other requirements meet the relevant regulations of 2010 editions relevant medicinal tea of the Pharmacopoeia of the People's Republic of China.
Embodiment 5: the preparation of blood sugar lowering concentrated pill
Get ginsenoside Rb3 1g among the embodiment 1, cyclodextrin 20g, carboxymethyl cellulose is an amount of, and mixing is pulverized, pure water, the general ball of making, every contains ginsenoside Rb3 1%-10% (g/g).Other requirements meet the relevant regulations of 2010 editions relevant pills of the Pharmacopoeia of the People's Republic of China.
Embodiment 6: the preparation of hypoglycemic granule agent
Get embodiment 1 ginsenoside Rb3 10g, soluble starch 200g, Icing Sugar is an amount of, magnesium stearate 3g, mixing, system granule, the packing of composite aluminium plastic bag, loading amount 5.0g/ bag.Other requirements meet the relevant regulations of 2010 editions relevant granules of the Pharmacopoeia of the People's Republic of China.
Embodiment 7: the preparation of cholesterol reducing tablet
Get the pure product 2g of embodiment 1 ginsenoside Rb3, cyclodextrin 20g, dry starch is an amount of, and carboxymethyl cellulose is an amount of, and Pulvis Talci is an amount of, and sucrose is an amount of.Sucrose and Pulvis Talci are used for sugar coating, mixing, and pelletizing press sheet makes the heavy 0.50g of tablet, contains ginsenoside Rb3 1%-5% (g/g).Other requirements meet the relevant regulations of 2010 editions relevant tablets of the Pharmacopoeia of the People's Republic of China.
Embodiment 8: the preparation of obesity tablet
Get embodiment 1 ginsenoside Rb3 component 10g (wherein contain ginsenoside Rb3 1g-5g, other has other constituent of ginseng), cyclodextrin 200g, dry starch is an amount of, and carboxymethyl cellulose is an amount of, and Pulvis Talci is an amount of, and sucrose is an amount of.Sucrose and Pulvis Talci are used for sugar coating, mixing, and pelletizing press sheet makes the heavy 0.50g of tablet, contains ginsenoside Rb3 1%-5% (g/g).Other requirements meet the relevant regulations of 2010 editions relevant tablets of the Pharmacopoeia of the People's Republic of China.
Embodiment 9: the preparation of obesity powder
Get Radix Ginseng (perhaps Radix Panacis Quinquefolii, Radix Notoginseng) the ginsenoside Rb3 component 10g among the embodiment 1, starch 200g, calcium phosphate 5g, lactose, food coloring are an amount of, and mixing sieves, divided dose, packing.Other requirements meet the relevant regulations of 2010 editions relevant tablets of the Pharmacopoeia of the People's Republic of China.
Claims (2)
1. ginsenoside Rb3 compositions, based on ginsenoside Rb3, be equipped with 1～5 kind of adjuvant in carrier, excipient, sweeting agent, adhesive, correctives, antiseptic, dispersant, surfactant, the coloring agent, the tablet of making, pill, granule, electuary, capsule and powder; Wherein in tablet, pill, the capsule every dose contain ginsenoside RB31～10wt%; Granule, electuary and powder are packed, every bag 5.0 gram;
Described ginsenoside Rb3 use following method extract and:
The stem and leaf crude extract of Radix Panacis Quinquefolii, Radix Ginseng or Radix Notoginseng, separate with the NKA-12 macroporous resin water-soluble back, and ethanol water gradient elution obtains panaxatriol's saponins and panoxadiol's saponins, panoxadiol's saponins with silicagel column repeatedly chromatography obtain the pure product of ginsenoside Rb3, purity 98%.
2. ginsenoside Rb3 compositions is preparing the application for the treatment of diabetes medicament.
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