CN102626438B - Antidiabetic medicament and preparation method - Google Patents
Antidiabetic medicament and preparation method Download PDFInfo
- Publication number
- CN102626438B CN102626438B CN201110175246.9A CN201110175246A CN102626438B CN 102626438 B CN102626438 B CN 102626438B CN 201110175246 A CN201110175246 A CN 201110175246A CN 102626438 B CN102626438 B CN 102626438B
- Authority
- CN
- China
- Prior art keywords
- extract
- caragana
- mas
- group
- preparation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Landscapes
- Medicines Containing Plant Substances (AREA)
Abstract
Disclosed are a diabetic-treating medicament and a preparation method, which comprises the following steps of: extracting Caragana opulens medicinal material by an ethyl acetate thermal reflux extraction method to obtain an extract product, and further purifying the obtained medicinal active site and a pharmaceutically acceptable conventional excipient by an alkali-solution and acid-isolation method and HPD-100 macroporous resin separation process to prepare an oral preparation. The content of an effective component total organic acid contained in the medicinal active site reaches up to more than 50%. It is proved through pharmacological experiments that the antidiabetic medicament has a significant hyperglycemic function and can be used as a medicament to treat or prevent type 2 diabetes and its complication; the medicament takes action in gastrointestinal tract and is convenient to take; By the adoption of the ethyl acetate thermal reflux extraction method and the alkali-solution and acid-isolation method and with the combination of the macroporous resin separation technology, total organic acid of Caragana opulens with high content of effective component can be obtained, and the preparation method has advantages of good curative effect, small drug dosage and little side effect, convenient production, little pollution and the like.
Description
Technical field
The present invention relates to treat the Chinese medicine of diabetes.
Background technology
In recent years, diabetes prevalence and mortality rate have obvious ascendant trend, and according to estimates, there is diabetic 1.5 hundred million in the whole world at present, within 2025, will reach 300,000,000, and the existing diabetics 3,000 ten thousand of China, will reach 5,000 ten thousand for 2025.Along with the increase of diabetics prevalence, complication, if scheming infarction, renal failure, uremia, blind etc. sickness rate are also along with raising, will seriously jeopardize patient's life, and Ci Bing China has become the third commonly encountered diseases after cardiovascular disease, tumor.And the medicine of diabetes is chemical synthetic drug mostly, although there is certain effect, exist action target spot single, easily bounce-back after drug withdrawal, the shortcoming that side effect is many.Yet much Chinese medicine has good hypoglycemic activity equally, its mechanism of action is many-sided.Special significant be to have much blood sugar lowering Chinese medicine to there is dual regulation, and Chinese medicine is hypoglycemic also has blood fat reducing concurrently simultaneously, anticoagulant and reduce viscosity of blood improves the multiple effects such as blood circulation.Therefore, Chinese medicine can not only blood sugar lowering and its complication is had to significant specific aim and preventive and therapeutic effect, filters out efficient, low toxicity, can prevent and treat again the medicine of complication from Chinese medicine, has very large advantage and exploitation is worth.
Sweet illiteracy caragana (Caragana opulens) is pulse family Caragana shrub, extensively originate in the ground such as Gansu, the Inner Mongol, that Tibetan medicine is looked into one of former plant of agate, cool in nature, bitter in the mouth, can be clear various muscle heat and arteries and veins are warm, can arrange disorders of meridian and have emetic action, its stem branch micromicro expelling wind and activating blood circulation, pain relieving diuresis, QI invigorating kidney tonifying.Up to the present, have no the modern study to this medical material chemistry and pharmacology.
Summary of the invention
The object of this invention is to provide a kind of medicine for the treatment of diabetes.
Another object of the present invention is to provide the preparation method of this antidiabetic medicine.
The present invention is a kind of antidiabetic medicine and preparation method, getting the sweet illiteracy caragana of pulse family population pattern change extracts and obtains extract with organic solvent water bath reflux method, further with the medical active position of alkali extraction and acid precipitation method and macroporous resin partition method purification gained and the oral formulations that pharmaceutically acceptable conventional adjuvant is made, in the dry product of said medical active position, by weight percentage, containing effective ingredient total organic acids, be greater than 50%;
The step of its preparation method is:
(1) getting the sweet illiteracy caragana of pulse family population pattern change plant is raw material, adopt organic solvent water bath reflux method, extraction solvent is ethyl acetate, extracts solid-to-liquid ratio 1: 15, extract temperature and be under the condition of 80 ℃ and extract 2～4 hours, evaporated under reduced pressure reclaims organic solvent and obtains extract;
(2) by extract obtained, further by alkali extraction and acid precipitation method, purify, be first dissolved in the 0.5M sodium hydroxide solution of 10 times of amounts, after standing over night, filter, filtrate is used 1M hydrochloric acid solution adjust pH to 2～3, then by the ethyl acetate of 2 times of amounts, extract 2 times, evaporated under reduced pressure obtains extractum
(3) by macroporous resin partition method, purify, first with washing with 30% alcoholic solution after HPD-100 type macroporous resin adsorption, then with 70% ethanol, resolve, evaporated under reduced pressure, obtain medical active position (being called for short MAS), get it filled and add pharmaceutically acceptable conventional adjuvant by conventional method of Chinese medicinal, make oral formulations or medical active position is undertaken adding pharmaceutically acceptable conventional adjuvant to make oral formulations by conventional method of Chinese medicinal after enclose by inclusion agents again with active site.
Sweet illiteracy caragana medical active position involved in the present invention is fat-soluble material, therefore pharmaceutical dosage form be take, prepares soft capsule as best, and the conventional adjuvant while preparing soft capsule has vegetable oil and PEG-4000; While preparing hard capsule, drop pill, tablet, with medical active position, first by inclusion agents, carry out enclose and make micro-rubber powder, then add pharmaceutically acceptable conventional adjuvant to be prepared into oral formulations by conventional method of Chinese medicinal.As the capsule powder of getting after enclose while preparing hard capsule, tablet adds adjuvant micropowder silica gel, starch, lactose, microcrystalline Cellulose etc., while preparing drop pill, add adjuvant PEG-4000, Polyethylene Glycol-600, plant wet goods.Inclusion agents is beta-schardinger dextrin-or modified starch, polishing while adopting beta-cyclodextrin inclusion compound, concrete grammar is: get the beta-schardinger dextrin-that weight is 10 times of amounts of active site, join in the beta-schardinger dextrin-water of 30 times of amounts, insert in colloid mill and grind evenly, then active site is dissolved in to 1 times of amount dehydrated alcohol, adds in colloid mill, grind 30 minutes, incline and lapping liquid, filter, filter cake is washed 3 times with a small amount of dehydrated alcohol, and vacuum drying or spraying are dry.
Oral dose when product provided by the invention is used for the treatment of diabetes, recommends in MAS (sweet illiteracy caragana extract is through the active site of alkali extraction and acid precipitation method and macroporous resin partition method purification gained) to take for each person every day 60-240mg.
The invention has the beneficial effects as follows: 1. the medicine that provided has remarkable anti-diabetic function, can be used for treating diabetes; 2. medicine is with gastrointestinal administration, and product forms is conveniently taken; Preparation method adopt organic solvent water bath reflux method and alkali extraction and acid precipitation method have convenient for production, pollute the advantages such as little.The present invention provides new selection by the medicine for treatment for diabetes.In preparation technology, adopted macroporous resin isolation technics, obtained the high sweet illiteracy caragana total organic acids of active constituent content, had good effect, dosage is little, the advantage that side effect is little.
Accompanying drawing explanation
Fig. 1 is the potentiometric titration curve figure of vanillic acid reference substance, and vertical coordinate is the ratio of potential change amount (Δ E) and volume change (Δ V), and abscissa is the volume averaging value (V) of the sodium hydroxide solution that spends before and after potential change.Potential change while having reacted sodium hydroxide solution titration vanillic acid in figure.
Fig. 2 is the potentiometric titration curve figure of total organic acids in sweet illiteracy caragana, vertical coordinate is the ratio of potential change amount (Δ E) and volume change (Δ V), and abscissa is the volume averaging value (V) of the sodium hydroxide solution that spends before and after potential change.Potential change while having reacted the sweet illiteracy caragana of sodium hydroxide solution titration extract in figure.
Fig. 3 is the curve chart of sweet illiteracy caragana extract MAS on alpha-glucosidase activity impact, and vertical coordinate is inhibition of enzyme activity rate, and abscissa representative sample suppresses the logarithm value of final concentration (μ g/ml).Sweet illiteracy caragana extract MAS and acarbose in figure, have been compared active to the inhibition of alpha-glucosidase when variable concentrations.
The specific embodiment
The preparation of embodiment 1, plant extract
1 potentiometric determination total organic acids content
(1) constant-current titration of vanillic acid reference substance
Vanillic acid 95% alcoholic solution of accurately drawing 25mL 0.0103mol/L, in 100mL beaker, adds 95% ethanol 15mL and shakes up, and connects potentiometer, magnetic stirring apparatus and solution to be measured, with the titration of 25mL burette splendid attire 0.051mol/L sodium hydroxide solution.Drip the sodium hydroxide volume that spends in time recording titration process and corresponding potential changing value thereof, carry out blank correction simultaneously.The ratio of potential change amount (Δ E) and volume change (Δ V) of take is vertical coordinate, and the volume averaging value (V) of the sodium hydroxide solution spending before and after potential change is abscissa, the results are shown in Figure 1.As shown in Figure 1, the maximum of curve is exactly titration end-point, and consumption alkali number is 5.05mL.By formula (V
alkali=C
acidv
acid/ C
alkali) calculation consumption alkali number is 5.05mL, the two matches, and curve display titration end-point is very obvious, illustrates that potentiometric titration surveys total organic acids content and have accuracy and susceptiveness.
(2) constant-current titration of total organic acids in sweet illiteracy caragana
In sweet illiteracy caragana, total organic acids content is in vanillic acid.Accurately take the about 240mg of sweet illiteracy caragana total acid extract in 100mL beaker, with 95% dissolve with ethanol solution, make test liquid, the making of titration curve is identical with the making of vanillic acid titration curve, the results are shown in Figure 2.As shown in Figure 2, the maximum of curve is exactly titration end-point, and consumption alkali number is 4.35mL.Curve display titration end-point is very obvious, illustrates that it is feasible that potentiometric titration is surveyed total organic acids content in sweet illiteracy caragana.
(3) calculating of total organic acids (vanillic acid meter) yield in sweet illiteracy caragana:
The volume of the sodium hydroxide that V---titration consumes, unit is: L
C---the concentration of titration sodium hydroxide used, unit is: mol/L
W---organic acid yield, unit is: %
The quality of M---medical material, unit is: g
2, the preparation of sweet illiteracy caragana crude extract
Choosing ethyl acetate is solvent, according to the influence factor who extracts, selected solvent extraction consumption 12-18 doubly, extraction time 2.5-3.5 hour, 1-3 3 factors of extraction time, each factor is got 3 levels, fixed extraction temperature is 80 ℃, employing L
9(3
4) orthogonal table Optimized Extraction Process condition, gauge outfit designs as table 1, and experimental data is in Table 2.Calculate by analysis the process conditions of selected optimum extraction.
Table 1 factor level table
Table 2 orthogonal experiments table
Experimental result by upper (table 2) can be found out, at L
9(3
4) total organic acids extraction time in orthogonal experiment three factors is more obvious on experimental result impact, it is the principal element that affects total organic acids yield, organic acid extraction ratio is had a significant effect, and other factors are relatively little on the impact of total organic acids yield in test. therefore each factor affects the primary and secondary relation of total organic acids extraction ratio, be C>A>B, optimised process is A
2b
3c
3.Optimised process concrete operations: sweet illiteracy caragana coarse powder 10Kg, with 15 times of volumes of acetic acid ethyl esters reflux, extract, at 80 ℃, extraction time is 3.5 hours, and extraction time is 3 times, and merge extractive liquid, is evaporated to the dry extractum 1 that obtains with Rotary Evaporators.
3, the preparation of sweet illiteracy caragana total organic acids
The sodium hydroxide solution that is 0.5M by the molar concentration of 20 times of amounts of gained extractum 1 use dissolves, after standing over night, filter, filtrate, with 1M hydrochloric acid solution adjust pH to 2～3, then extracts 2 times by the ethyl acetate of 2 times of amounts, and evaporated under reduced pressure obtains extractum 2, HPD-100 macroporous resin will be crossed after gained extractum 2 use 30% dissolve with ethanols, ethanol with 70% after absorption is completely resolved, and by desorbed solution evaporated under reduced pressure, obtains extractum 3, be medical active position (being called for short MAS), yield is 0.21%.By potentiometric titration, detect, wherein total organic acids content is 64.5%.
(1) enclose of MAS
1, in 1Kg beta-schardinger dextrin-, add 30L water, insert in colloid mill and grind evenly.Then MAS100g is dissolved in to 100ml dehydrated alcohol, adds in colloid mill, grind 30 minutes.Incline and lapping liquid, sucking filtration.Filter cake is washed 3 times with a small amount of dehydrated alcohol, and vacuum drying or spraying are dried to obtain 1098g MAS clathrate.
2, in 500g modified starch N-LOK, add 15L water, insert in colloid mill and grind evenly.Then MAS50g is dissolved in to 50ml dehydrated alcohol, adds in colloid mill, grind 30 minutes.Incline and lapping liquid, sucking filtration.Filter cake is washed 3 times with a small amount of dehydrated alcohol, and vacuum drying or spraying are dried to obtain 547g MAS clathrate.
(2) preparation of medicine
1, the preparation of soft capsule
Content: get MAS60g, vegetable oil: 300g, glycerol: 20g, Tween 80: 0.1g mix homogeneously.
Get content and be pressed into 1000 soft capsules with rotation rolling capsule machine.
2, the preparation of hard capsule
Get MAS clathrate 220g, cross 80 mesh sieves, add appropriate 10% starch slurry to make soft material, cross 14 order nylon mesh and granulate, 60 ℃ are dried to obtain granule, insert capsulae vacuus, make altogether 1000 of hard capsules.
3, the preparation of tablet
Get MAS clathrate 220g and starch 85g, dextrin 66g, sucrose 10g mix granulation.60 ℃ following dry, adds Pulvis Talci 5g and magnesium stearate 1g to mix, and is pressed into 1000.
4, the preparation of drop pill
Get 180g PEG-4000 heating and make into molten condition, add 60g MAS, mix, splash in cooling liquid paraffin, routine is made 1000 drop pill.
Below use according to the sweet illiteracy caragana of embodiment 1 gained extract MAS and carry out in vitro tests, pharmacodynamics test and toxicologic study
One, sweet illiteracy caragana extract MAS is to the inhibiting in vitro tests of alpha-glucosidase
1, material
Sweet illiteracy caragana extract MAS (pressing embodiment 1 preparation), acarbose (Bayer Bitterfeld GmbH medicines and health protection company limited), 4-Nitrobenzol-α-D-pyranglucoside (pNPG, Sigma company), alpha-D-glucose glycosides enzyme (Sigma company).
2, method
Sweet illiteracy caragana extract MAS and acarbose are dissolved in respectively to DMSO, being made into concentration is the sample liquid of 5mg/ml, then use phosphate buffer (10mmol/L, pH=6.8) dilution is the liquid to be measured of variable concentrations (10-500 μ g/ml), getting 120 μ l phosphate buffers and 20 μ l sample liquid and 0.75U/ml glucosidase 10 μ l mixes, in 4 ℃, cultivate 20min, add 10mmol/L pNPG (phosphate buffer dissolving) 50 μ l, in 37 ℃ of incubation 20min, under 405nm wavelength, measure OD value, more than react on 96 orifice plates and complete, reaction cumulative volume is 200 μ l.Each test sample is done 3 multiple holes simultaneously, averages, and 3 repeated experiments.Acarbose, as the positive control of this law, is set blank and negative control simultaneously.Calculate the suppression ratio of enzymatic activity: suppression ratio=((sample-feminine gender)/(blank-feminine gender)) * 100%.Logarithm with sample final concentration (μ g/ml) is done abscissa, and suppression ratio is done vertical coordinate mapping, and result as shown in Figure 3.
By Fig. 3 result, can be found out, sweet illiteracy caragana extract MAS has remarkable inhibitory action to alpha-glucosidase in vitro, and effect is suitable with acarbose.
Two, the Pharmacodynamic test of active extract of sweet illiteracy caragana extract MAS
(1) impact of sweet illiteracy caragana extract MAS on normal mouse starch-bearing carbohydrate tolerance
1, material
Sweet illiteracy caragana extract MAS (pressing embodiment 1 preparation), acarbose (production of Bayer Bitterfeld GmbH medicines and health protection limit company), Kunming mouse (preclinical medicine institute of Lanzhou University Experimental Animal Center provides), blood glucose monitoring system (Beijing Yi Cheng Bioisystech Co., Ltd), blood glucose examination bar (Beijing Yi Cheng Bioisystech Co., Ltd).
2, method
Get 50 of healthy adult male mices, fasting 2 hours, is divided into 5 groups at random: high, normal, basic three dosage group (50mg/Kg of Normal group, acarbose group and sweet illiteracy caragana extract MAS, 100mg/Kg, 200mg/Kg), 10 every group, the blood glucose there was no significant difference that each is organized.Respectively give corresponding dosage once after, with 5g/kg dosage starch gavage, give starch after 3 hours, mouse blood sugar is surveyed in eye socket blood sampling.Result is as shown in table 3:
The impact of the sweet illiteracy caragana of table 3 extract MAS on normal mouse blood sugar
| Group | Dosage (mg/Kg) | Blood glucose value (mmol/L) |
| Normal group | - | 6.92±1.13 |
| MAS group (low) | 50 | 6.76±1.32 |
| MAS group (in) | 100 | 5.98±0.82* |
| MAS group (height) | 200 | 5.72±0.53** |
| |
20 | 5.16±0.64** |
Note: with Normal group comparison, * p<0.05, * * p<0.01.
As can be seen from Table 3: the post-prandial glycemia that high, the middle dosage group of sweet illiteracy caragana extract MAS all can significantly suppress normal mouse raises (p<0.05 or p<0.01), and effect and the acarbose of its high dose group are suitable.
(2) sweet illiteracy caragana extract MAS is on alloxan induction diabetic mice starch-bearing carbohydrate tolerance impact test
1, material
Sweet illiteracy caragana extract MAS (pressing embodiment 1 preparation), Kunming mouse (preclinical medicine institute of Lanzhou University Experimental Animal Center provides), alloxan (production of Sigma company), glibenclamide (Shaanxi Sen Fu Bioisystech Co., Ltd), blood glucose monitoring system (Beijing Yi Cheng Bioisystech Co., Ltd), blood glucose examination bar (Beijing Yi Cheng Bioisystech Co., Ltd).
2, method
Get body weight 22-26g Kunming mouse, lumbar injection 200mg/kg alloxan, after 72 hours, measure blood glucose, choose the mice of blood glucose value more than 11.1mmol/L, by 10 every group, male and female half and half are divided into 6 groups, be diabetic model group, acarbose group, sweet illiteracy caragana extract MAS high dose group, middle dosage group, low dose group, acarbose dosage is 20mg/kg (body weight), and sweet illiteracy caragana extract MAS high dose group is that 200mg/kg, middle dosage group are that 100mg/kg, low dose group are 50mg/kg; Measure fasting glucose (0min), administration is (model group gavage is with normal saline) after ten minutes, and each organizes equal gavage with starch 5g/kg, respectively at 30min, 60min, 90min, 120min, gets eye socket rear vein beard blood, with blood glucose examination bar, measure blood sugar content, the results are shown in Table 4.
The impact of the sweet illiteracy caragana of table 4 extract MAS on alloxan induction diabetic mice starch-bearing carbohydrate tolerance
Note: with model group comparison, * p<0.05, * * p<0.01.
As can be seen from Table 4: diabetic mice is after giving starch, and blood glucose value (model group) obviously raises.Sweet illiteracy caragana extract MAS high dose group all significantly suppresses the blood sugar increasing of diabetic mice at each time point, and its effect is suitable with acarbose; Compare with model group, in MAS, low dose group all has certain inhibitory action to the blood sugar increasing of each time point diabetic mice, but effect is not as good as MAS high dose group.
(3) impact of sweet illiteracy caragana extract MAS on the blood glucose of diabetic mice due to streptozotocin
1, material
Sweet illiteracy caragana extract MAS (pressing embodiment 1 preparation), Kunming mouse (preclinical medicine institute of Lanzhou University Experimental Animal Center provides), glibenclamide (Shaanxi Sen Fu Bioisystech Co., Ltd), streptozotocin (production of Sigma company), blood glucose monitoring system (Beijing Yi Cheng Bioisystech Co., Ltd), blood glucose examination bar (Beijing Yi Cheng Bioisystech Co., Ltd).
2, method
Get 80 of healthy male mices, choose at random 10 as Normal group, all the other 70 fasting 12 hours, press the dosage lumbar injection streptozotocin of 60mg/kg body weight, Normal group injection citrate buffer solution, within 1 week after modeling, measure fasting blood sugar, the mice that screening blood glucose value surpasses 11.1mmol/L is experimental model Mus.50 diabetes experimental model Mus are divided into diabetic model group at random, glibenclamide group (gavage 20mg/kg), the basic, normal, high dosage group of sweet illiteracy caragana extract MAS (is respectively 50,100,200mg/kg).Normal group and diabetic model group gavage normal saline, successive administration 3 weeks.After last administration, fasting is 12 hours, and eye socket is got blood, surveys change of blood sugar value, the results are shown in Table 5.
The impact of the sweet illiteracy caragana of table 5 extract MAS on blood glucose in diabetic mice due to streptozotocin
| Group | Dosage (mg/kg) | Blood glucose value (mmol/L) |
| Normal group | - | 4.7±0.9** |
| Model control group | - | 15.4±3.8 |
| |
20 | 9.1±4.9** |
| MAS group (height) | 200 | 9.3±4.5** |
| MAS group (in) | 100 | 10.6±5.3** |
| MAS group (low) | 50 | 12.4±6.1* |
Note: with model control group comparison, * p<0.05, * * p<0.01.
As can be seen from Table 5: compare with model group, the high, medium and low dosage group of sweet illiteracy caragana extract MAS all can significantly reduce the blood glucose value (p<0.05 or p<0.01) of diabetic mice, and effect and glibenclamide high, middle dosage group are suitable.
(4) impact of sweet illiteracy caragana extract MAS on the blood glucose in diabetic mice of alloxan induction
1, material
Sweet illiteracy caragana extract MAS (pressing embodiment 1 preparation), Kunming mouse (preclinical medicine institute of Lanzhou University Experimental Animal Center provides), glibenclamide (Shaanxi Sen Fu Bioisystech Co., Ltd), alloxan (production of Sigma company), blood glucose monitoring system (Beijing Yi Cheng Bioisystech Co., Ltd), blood glucose examination bar (Beijing Yi Cheng Bioisystech Co., Ltd).
2, method
80 of male mice in kunming, before experiment, fasting is 12 hours, chooses at random 10 as Normal group, all the other lumbar injection 200mg/kg alloxan, after 72 hours, survey blood glucose value, choosing wherein 50 blood glucose value >11.1mmol/L persons as diabetic mice.Diabetic mice is divided into high, medium and low three the dosage group (50mg/Kg of sweet illiteracy caragana extract MAS at random, 100mg/Kg, 200mg/Kg), model control group, glibenclamide positive controls (20mg/Kg), make there was no significant difference between each blood glucose value mean of organizing, 10 every group.Each organizes gastric infusion or normal saline every day, and continuous 3 weeks, in last fasting, within 12 hours, after administration, within 1 hour, from mouse orbit, get blood again, with blood glucose examination bar, measure blood sugar content, the results are shown in Table 6.
The impact of the sweet illiteracy caragana of table 6 extract MAS on the blood glucose in diabetic mice of alloxan induction
| Group | Dosage (mg/kg) | Blood glucose value (mmol/L) |
| Normal group | - | 5.4±0.8** |
| Model control group | - | 19.3±2.6 |
| |
20 | 13.1±1.9** |
| MAS group (height) | 200 | 14.3±2.5** |
| MAS group (in) | 100 | 15.6±3.3** |
| MAS group (low) | 50 | 17.4±3.8* |
Note: with model control group comparison, * p<0.05, * * p<0.01.
As shown in Table 6, administration group blood glucose is compared remarkable reduction (P<0.01) with model control group, and blood glucose declines and sweet illiteracy caragana extract MAS is dose dependent, and effect and the glibenclamide of high dose group are suitable.
(5) impact of sweet illiteracy caragana extract MAS on the diabetic mice liver glycogen content of alloxan induction
1, material
Sweet illiteracy caragana extract MAS (pressing embodiment 1 preparation), Kunming mouse (preclinical medicine institute of Lanzhou University Experimental Animal Center provides), glibenclamide (Shaanxi Sen Fu Bioisystech Co., Ltd), alloxan (production of Sigma company).
2, method
80 of male mice in kunming, before experiment, fasting is 12 hours, chooses at random 10 as Normal group, all the other lumbar injection 200mg/kg alloxan, after 72 hours, measure blood glucose value, choosing wherein 50 blood glucose value >11.1mmol/L as diabetic mice.Diabetic mice is divided into high, medium and low three the dosage group (50mg/Kg of sweet illiteracy caragana extract MAS at random, 100mg/Kg, 200mg/Kg), model control group, and glibenclamide positive controls (20mg/Kg), make there was no significant difference between each blood glucose value mean of organizing, 10 every group.Each organizes gastric infusion or normal saline every day, continuous 21 days, in last administration after 1 hour disconnected marrow put to death, by sulphuric acid-fear ketone method, measure liver glycogen content, result is as shown in table 7:
The impact of the sweet illiteracy caragana of table 7 extract MAS on the diabetic mice liver glycogen content of alloxan induction
| Group | Dosage (mg/kg) | Liver glycogen content (mg/g) |
| Normal group | - | 35.3±1.8** |
| Model control group | - | 17.6±1.6 |
| |
20 | 25.1±4.7** |
| MAS group (height) | 200 | 24.3±3.5** |
| MAS group (in) | 100 | 23.6±3.3** |
| MAS group (low) | 50 | 21.4±2.1* |
Note: with model control group comparison, * p<0.05, * * p<0.01.
As can be seen from Table 7: compare with model group, the high, medium and low dosage group of sweet illiteracy caragana extract MAS all can significantly reduce the liver glycogen consumption of diabetic mice, and effect and glibenclamide high, middle dosage group suitable.
(6) diabetic mice blood insulin and the islet tissue morphologic impact of sweet illiteracy caragana extract MAS on alloxan induction
1, material
Sweet illiteracy caragana extract MAS (pressing embodiment 1 preparation), Kunming mouse (preclinical medicine institute of Lanzhou University Experimental Animal Center provides), alloxan (production of Sigma company), fears ketone reagent (5-linked chemical plant, Shanghai).
2, method
60 of male mice in kunming, before experiment, fasting is 12 hours, chooses at random 10 as Normal group, all the other lumbar injection 200mg/kg alloxan, after 72 hours, measure blood glucose value, choosing wherein 30 blood glucose value >11.1mmol/L persons as diabetic mice.Diabetic mice is divided into sweet illiteracy caragana extract MAS height two dosage groups (50mg/Kg, 200mg/Kg) and model control group, 10 every group at random.Each organizes gastric infusion or normal saline every day, continuous 21 days, in last administration after 1 hour disconnected marrow put to death, from each group, get at random respectively 3 mice pancreatic afterbody tissues, use Gormori aldehyde-fuchsin and H.E colouring method, observation islet tissue.Observed result is in Table 8:
The morphological change of table 8 islet tissue
As can be seen from Table 8: sweet illiteracy caragana extract MAS high dose group has significant repair to the insulin secreting cells being destroyed by alloxan, and has increased the secretory granule of beta Cell of islet.Compare with model group, MAS low dose group has certain repair to the insulin secreting cells being destroyed by alloxan, but effect is not as good as MAS high dose group.
(7) impact of sweet illiteracy caragana extract MAS on normal mouse blood sugar
1, material
Sweet illiteracy caragana extract MAS (pressing embodiment 1 preparation), Kunming mouse (preclinical medicine institute of Lanzhou University Experimental Animal Center provides), blood glucose monitoring system (Beijing Yi Cheng Bioisystech Co., Ltd), blood glucose examination bar (Beijing Yi Cheng Bioisystech Co., Ltd).
2, method
40 of normal mouses, male and female half and half, are divided into 4 groups at random: Normal group, gavage normal saline; High, medium and low dosage group (50mg/Kg, 100mg/Kg, 200mg/Kg) is the sweet illiteracy caragana of gavage extract MAS respectively, 1 1d, continuous use 3 weeks.3 hours posterior orbits of last administration are got blood, survey change of blood sugar value, the results are shown in Table 9.
The impact of the sweet illiteracy caragana of table 9 extract MAS on normal mouse blood sugar
| Group | Dosage (mg/kg) | Blood glucose value (mg/g) |
| Normal group | - | 5.3±1.8 |
| MAS group (height) | 200 | 4.3±1.5* |
| MAS group (in) | 100 | 4.6±1.3* |
| MAS group (low) | 50 | 5.1±2.1 |
Note: with model control group comparison, * p<0.05, * * p<0.01.
As can be seen from Table 9: with Normal group comparison, the high, medium and low dosage group of sweet illiteracy caragana extract MAS all can make the blood glucose value of normal mouse reduce, and the effect of high, middle dosage group significantly (p<0.05).
Three, acute toxicity testing
1, material
Sweet illiteracy caragana extract MAS (press embodiment 1 preparation), Kunming mouse (preclinical medicine institute of Lanzhou University Experimental Animal Center provides), 0.5% methylcellulose is received (medicine dissolution diluent).
2, method
Respectively get 20 of normal mouses, male and female half and half, receive sweet illiteracy caragana extract MAS are made into and can fill with concentration 0.5g/mL with 0.5% methylcellulose, with the administration of every mice 20ml/kg body weight, disposable gavage, the poisoning and death condition of mice in 14 days after observation administration.Toxic reaction primary part observation symptom, degree, toxic reaction initial time, persistent period and recovery time, cut open the dead white mice of inspection, records dead mouse reason.After within the 14th day, putting to death whole white mice, cut open inspection, the main organs such as perusal heart, liver, spleen, lungs, kidney.
3, result
After gastric infusion mice occur in various degree few moving, do not take food or the phenomenon such as few feed, but recovered successively after 2 hours, mice shows no obvious abnormalities reaction, freely, diet, feces and other situations be all without extremely in activity, hair color light next to the skin.After administration, 7 days and 14 days animal subject body weight are showed no significant change, cut open inspection after within the 14th day, putting to death whole mices, and the main organs no abnormality seens such as perusal heart, liver, spleen, lungs, kidney change.Result shows that mouse stomach administration is greater than 10g/Kg body weight to the maximum tolerated dose of sweet illiteracy caragana extract MAS, illustrates that this plant extract toxicity is extremely low or nontoxic.
By above-mentioned in vitro tests and pharmacodynamics test, prove that sweet illiteracy caragana extract MAS has the effect that reduces post-prandial glycemia, the activity of energy Inhibiting α-glucosidase, and basic avirulence, have the features such as evident in efficacy, cheap, safe and convenient to use.
Claims (5)
1. an antidiabetic medicine, it is characterized in that: get the sweet illiteracy caragana of pulse family population pattern change and extract and obtain extract with organic solvent water bath reflux method, further by alkali extraction and acid precipitation method and macroporous resin partition method, purify, the medical active position of gained and pharmaceutically acceptable conventional adjuvant are made oral formulations, in the dry product of said medical active position, by weight percentage, containing effective ingredient total organic acids, be greater than 50%;
The step of its preparation method is:
(1) getting the sweet illiteracy caragana of pulse family population pattern change plant is raw material, adopt organic solvent water bath reflux method, extraction solvent is ethyl acetate, extracts solid-to-liquid ratio 1: 15, extract temperature and be under the condition of 80 ℃ and extract 2～4 hours, evaporated under reduced pressure reclaims organic solvent and obtains extract;
(2) by extract obtained, further by alkali extraction and acid precipitation method, purify, be first dissolved in the 0.5M sodium hydroxide solution of 10 times of amounts, after standing over night, filter, filtrate is used 1M hydrochloric acid solution adjust pH to 2～3, then by the ethyl acetate of 2 times of amounts, extract 2 times, evaporated under reduced pressure obtains extractum
(3) with HPD-100 type macroporous resin, carry out separating-purifying, first with washing with 30% alcoholic solution after HPD-100 type macroporous resin adsorption, then with 70% ethanol, resolve, evaporated under reduced pressure, obtain medical active position, get it filled and add pharmaceutically acceptable conventional adjuvant by conventional method of Chinese medicinal, make oral formulations or medical active position is undertaken adding pharmaceutically acceptable conventional adjuvant to make oral formulations by conventional method of Chinese medicinal after enclose by inclusion agents again with active site.
2. antidiabetic medicine according to claim 1, is characterized in that: said oral formulations is hard capsule, or soft capsule, or drop pill, or tablet.
3. the preparation method of antidiabetic medicine claimed in claim 1, the steps include:
(1) getting the sweet illiteracy caragana of pulse family population pattern change plant is raw material, adopt organic solvent water bath reflux method, extraction solvent is ethyl acetate, extracts solid-to-liquid ratio 1: 15, extract temperature and be under the condition of 80 ℃ and extract 2～4 hours, evaporated under reduced pressure reclaims organic solvent and obtains extract;
(2) by extract obtained, further by alkali extraction and acid precipitation method, purify, be first dissolved in the 0.5M sodium hydroxide solution of 10 times of amounts, after standing over night, filter, filtrate is used 1M hydrochloric acid solution adjust pH to 2～3, then by the ethyl acetate of 2 times of amounts, extract 2 times, evaporated under reduced pressure obtains extractum
(3) with HPD-100 type macroporous resin, carry out separating-purifying, first with washing with 30% alcoholic solution after HPD-100 type macroporous resin adsorption, then with 70% ethanol, resolve, evaporated under reduced pressure, obtain medical active position, get it filled and add pharmaceutically acceptable conventional adjuvant by conventional method of Chinese medicinal, make oral formulations or medical active position is undertaken adding pharmaceutically acceptable conventional adjuvant to make oral formulations by conventional method of Chinese medicinal after enclose by inclusion agents again with active site.
4. the preparation method of antidiabetic medicine according to claim 3, is characterized in that: described inclusion agents is beta-schardinger dextrin-or modified starch N-LOK.
5. the preparation method of antidiabetic medicine according to claim 4, it is characterized in that: described active site by the method that inclusion agents is carried out enclose is: get the beta-schardinger dextrin-that weight is 10 times of amounts of active site, join in the water of beta-schardinger dextrin-consumption of 30 times of amounts, insert in colloid mill and grind evenly, then active site is dissolved in to one times of amount dehydrated alcohol, add in colloid mill, grind 30 minutes, incline and lapping liquid, filter, a small amount of absolute ethanol washing 3 times for filter cake, vacuum drying or spraying are dry.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110175246.9A CN102626438B (en) | 2011-06-22 | 2011-06-22 | Antidiabetic medicament and preparation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110175246.9A CN102626438B (en) | 2011-06-22 | 2011-06-22 | Antidiabetic medicament and preparation method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102626438A CN102626438A (en) | 2012-08-08 |
| CN102626438B true CN102626438B (en) | 2014-04-02 |
Family
ID=46584946
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201110175246.9A Expired - Fee Related CN102626438B (en) | 2011-06-22 | 2011-06-22 | Antidiabetic medicament and preparation method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102626438B (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1554626A (en) * | 2003-12-22 | 2004-12-15 | 复旦大学 | Eudesmane compounds, preparing method and use in preparing medicinal composition |
-
2011
- 2011-06-22 CN CN201110175246.9A patent/CN102626438B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1554626A (en) * | 2003-12-22 | 2004-12-15 | 复旦大学 | Eudesmane compounds, preparing method and use in preparing medicinal composition |
Non-Patent Citations (5)
| Title |
|---|
| 杨中铎等.锦鸡儿属植物化学成分及生物活性研究进展.《中成药》.2008,第30卷(第11期),1678-1681. |
| 甘蒙锦鸡儿化学成分研究;肖岸容;《中国优秀硕士学位论文全文数据库医药卫生科技辑》;20091130(第11期);E057-89 * |
| 肖岸容.甘蒙锦鸡儿化学成分研究.《中国优秀硕士学位论文全文数据库医药卫生科技辑》.2009,(第11期),E057-89. |
| 蒙秋霞.小叶锦鸡儿黄酮类化合物研究.《中国优秀博硕士学位论文全文数据库（硕士）农业科技辑》.2006,(第2期),D049-41. * |
| 锦鸡儿属植物化学成分及生物活性研究进展;杨中铎等;《中成药》;20081130;第30卷(第11期);1678-1681 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102626438A (en) | 2012-08-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102274244B (en) | Cassia bark polyphenol extract and preparation method and application thereof | |
| CN101032504B (en) | Application of liquiritin in medicines | |
| CN113143997A (en) | Application of mulberry extract in preparation of medicine for reducing animal weight | |
| CN103349671B (en) | A kind of resveratrol Spirulin composition and preparation thereof and method for making | |
| CN1965873B (en) | Chinese medicinal extract having blood sugar-lowering activity, its preparation process, composition and use | |
| CN103622946A (en) | Medical application of anhydroicaritin | |
| CN102872227A (en) | Gelan Xinning soft capsule for treating coronary disease and angina and preparation method thereof | |
| CN102716135B (en) | Lupenone prevents in preparation or treats the application in the product of diabetes | |
| AU2007204495B2 (en) | An extract of Xanthoceras sorbifolia Bunge and extraction and uses thereof | |
| CN103585192A (en) | Preparation method and application of Aleuritopteris argentea Fee extract | |
| CN103271963A (en) | Calliopsis elegans aqueous extract and medical application thereof | |
| CN1931233B (en) | Medicine composition of red sage and epimedium for treating cardiac and cerebral vascular diseases | |
| CN102626438B (en) | Antidiabetic medicament and preparation method | |
| CN107778340A (en) | (20S, 24R) 20,24 epoxy dammarane 3 β, 12 β, 25 triols and its application | |
| CN100534461C (en) | Pharmaceutical composition for treating diabetes and impaired glucose tolerance and preparation method thereof | |
| CN100563647C (en) | The Pharmaceutical composition of Herba Erigerontis and sodium tanshinon IIA silate injection | |
| CN101099754A (en) | Preparation method and application for pedunculoside II | |
| CN1923228B (en) | Pharmaceutical composition comprising notoginseng extract, Danshen extract and ligustrazine | |
| CN102824423B (en) | Medicinal composition comprising albiflorin and arctiin and application | |
| CN105663053A (en) | Sugar-free granules capable of promoting blood circulation and preparation method thereof | |
| CN105168300A (en) | Pharmaceutical composition for treating diabetes and preparation method thereof | |
| CN101185662A (en) | Method and use for preparing novel medicine for treating diabetes prepared from tuber fern | |
| CN1899410A (en) | Medicine for treating cardiovascular disease and its preparing method and quality control method | |
| CN103405724B (en) | A kind of Chinese medicine composition and preparation method thereof | |
| CN101406576A (en) | Chinese medicine for treating diabetes and preparation method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20140402 Termination date: 20150622 |
|
| EXPY | Termination of patent right or utility model |