CN1511576A - Medicinal composition for preventing and curing diabetes and its preparing method - Google Patents
Medicinal composition for preventing and curing diabetes and its preparing method Download PDFInfo
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Abstract
The present invention discloses one medicine composition for treating liver fibrosis. The medicine composition is prepared with 11 kinds of Chinese medicinal materials including astragalus root, kudzu vine root, Chinese yam, atractylodes rhizome, anemarrhena rhizome, etc. The medicine composition has excellent curative effect on diabetes.
Description
Technical field
The present invention relates to a kind of pharmaceutical composition and preparation method thereof, particularly relate to a kind of pharmaceutical composition of preventing and treating diabetes and preparation method thereof.
Background technology
Diabetes are endocrine and metabolic disorders diseases that a class is caused by multi-pathogenesis such as heredity, environment, immunity.Be characterized in chronic hyperglycemia, follow because of insulin secretion and/or effect the defective sugar, fat and the protein metabolism disorder that cause.Basic pathology physiology is absolute or relative property hypoinsulinism and glicentin activity increase caused metabolism disorder.Clinically polydipsia, polyphagia, polyuria occur, become thin, symptom such as fatigue, complication extensively and serious often can cause the heart, cerebrovascular, kidney, optical fundus, retina and nervous system lesion, ketoacidosis can take place, complication such as hyperosmolar coma when serious.Diabetes are chronic lifelong participation disease, in case take place, need take medicine all the life.Diabetes are commonly encountered diseases, frequently-occurring disease, and annual morbidity is 9,30/,100,000, and diabetes have the extension and the tendency of rejuvenation now, and every day is with 3500 people's speed increase with 1,250,000 every year for China's new trouble diabetes number
[1], the growth that this is surprising makes diabetes formally clamp-on the row of ten big representative national diseases.Diabetes are second killers in the modern disease, and its harm to human body is only second to cancer, cause serious body and mind infringement and burden on society.Therefore, actively develop the study on prevention of diabetes, become an important topic of current sanitary work.The treatment of diabetes purpose is to correct carbohydrate metabolism disturbance, promotes islet function to recover, and improves the insulin resistant state, and in time prevents and treats various complication.Doctor trained in Western medicine oral antidiabetic drug curative effect is affirmed, but antibody can appear in life-time service, causes secondary failure, and has side effect in various degree, as hypoglycemia, gastrointestinal upset etc.The insulin determined curative effect, but need drug administration by injection, to use inconvenience, and, its biological activity is reduced for a long time with easily producing antibody, the dosage increase then easily produces hyperinsulinemia and quickens the generation and the development of various chronic complicating diseases.The Chinese medicine document does not have " diabetes " name of disease, and is according to the clinical manifestation of diabetes, as " more than three " diseases such as polyuria, polyphagia, polydipsia, consistent with the Chinese medicine sick clinical manifestation of " quenching one's thirst ".TCM treatment of diabetes is based on organic conception, determination of treatment based on pathogenesis obtained through differentiation of symptoms and signs, at qi and blood prosperity and decline, wane and wax of yin and yang and mechanism of qi, expectorant stasis of blood situation, adjusts the human internal environment, improves patient's metabolism status and reaches therapeutic purposes.Though the Chinese medicine blood sugar decreasing effect than Western medicine a little less than, it is lasting but effect relaxes, can improve symptom, blood sugar lowering effectively, prevent and treat complication, disease controlling development and because many Chinese medicines have dual regulation, generally can not cause untoward reaction such as hypoglycemia, the irreplaceable advantage of Western medicine is arranged, be subjected to the attention of Chinese scholars day by day.Therefore, fully inherit motherland's medical science and prevent and treat the precious legacy of diabetes, outstanding tcm characteristic, bring into play its advantage, in conjunction with research and the Chinese medicine latest developments of modern medicine to diabetes, develop pure natural medical efficient, that have no side effect, have important theoretical meaning and value for clinical application, will produce good economic benefit and social benefit.The object of the invention is to provide a kind of pharmaceutical composition of preventing and treating diabetes and preparation method thereof.The object of the invention is to provide a kind of method of quality control.
Summary of the invention
The present invention seeks to be achieved through the following technical solutions, crude drug is by weight:
Radix Astragali 400-800 weight portion Radix Puerariae 300-500 weight portion Rhizoma Dioscoreae 300-500 weight portion
Rhizoma Atractylodis 200-400 weight portion Rhizoma Anemarrhenae 200-400 weight portion Radix Trichosanthis 200-400 weight portion
Radix Rehmanniae 300-500 weight portion Herba Portulacae 200-400 weight portion Radix Salviae Miltiorrhizae 200-400 weight portion
Ramulus Euonymi 200-400 weight portion Ramulus Mori 200-400 weight portion
The above-mentioned raw materials medicine can be made any clinical acceptable forms, as tablet, and capsule, granule, oral liquid, subcutaneous administration preparation, suppository etc.
Preparation of drug combination method of the present invention is: get Rhizoma Atractylodis, Radix Salviae Miltiorrhizae two flavors and doubly measured the 85-95% alcohol reflux 1-2 hour with 4-8, filter, medicinal residues and Radix Puerariae merge, with 60-80% alcohol reflux 2-4 time, each 4-7 doubly measures, and extracts respectively 0.5-2 hour, filter, merging filtrate, decompression recycling ethanol, water liquid is standby; Get eight flavor medicines such as the Radix Astragali and decoct with water 2-3 time, add 8-12 times of water at every turn, decocted 1-2 hour, collecting decoction filters, and filtrate is concentrated into 1.10 ~ 1.12/60 ℃ of relative densities, be blended into above-mentioned alcohol extraction concentrated solution, spray drying is made clinical acceptable forms, as tablet, capsule, granule, oral liquid, the subcutaneous administration preparation, suppository etc.
Comprise in the method for quality control of granule of the present invention and differentiating and/or assay that discriminating comprises one or more in the following method:
Differentiate that a. gets this product powder 2g, add methanol 40ml supersound extraction 20-40 minute, filter, filtrate evaporate to dryness, residue add water 15ml makes dissolving,
Divide 2-4 extraction with ether, each 15-30ml discards ether solution, the water intaking layer divides 2-4 extraction with water-saturated n-butanol, and each 15-30ml merges n-butanol layer 0.5-2%, sodium hydroxide solution divides 2-3 washing, each 10-20ml discards alkali liquor, n-butyl alcohol with n-butyl alcohol saturated be washed to neutrality, get n-butyl alcohol liquid, be concentrated into driedly, residue adds water 5ml makes dissolving, is added on the D that has handled well
101Macroporous resin column, difference water 40-60ml, 30-50% ethanol 20-40ml, 60-80% ethanol 40-60ml eluting is collected the 60-80% ethanol elution, and evaporate to dryness, residue add methanol 2ml makes dissolving, as need testing solution.Other gets the astragaloside reference substance, adds methanol and makes solution that every 1ml contains 1mg product solution in contrast.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B); Draw need testing solution 8 μ l, astragaloside reference substance 4 μ l, put respectively on same silica gel g thin-layer plate, with 11-14: 6-9: lower floor's solution that 1-2 chloroform-methanol-water is placed below 10 ℃ is developing solvent, launches, and takes out, dry, spray is with 10 ethanol solution of sulfuric acid, and 100-110 ℃ is heated 5-8min, and is clear to the speckle colour developing.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
B. get this product 3g, add ethanol 30ml, supersound extraction 20-40 minute, filter, filtrate is concentrated into 2ml, as need testing solution.Other gets Rhizoma Atractylodis control medicinal material 2g, shines medical material solution in pairs with legal system, according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B) test, draw need testing solution 4 μ 1, control medicinal material solution 2 μ l put respectively on same silica gel G plate, with petroleum ether 60-90 ℃ be developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid of 5% vanillin, and it is clear that hot blast blows to the speckle colour developing, in the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the speckle of same color.
C. get and differentiate that need testing solution is as need testing solution under the b item, other gets Radix Salviae Miltiorrhizae control medicinal material 2g, adds ethanol 30ml, and supersound extraction 20-40 minute, filter, filtrate is concentrated into 2ml, in contrast medical material solution.Get the tanshinone reference substance again, add ethyl acetate and make the solution that every 1ml contains 2mg.According to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2000) test, draw each 4 μ 1 of above-mentioned three kinds of solution, reference substance solution 2 μ l, put respectively on same silica gel g thin-layer plate, with 16-19: 1-4 benzene-ethyl acetate is developing solvent, launches, take out, dry.In the test sample chromatograph, with control medicinal material chromatograph and reference substance chromatograph relevant position on, show the speckle of same color.
D. get this product 2g, use methanol 20ml ultrasonic 20-40 minute, filter, filtrate evaporate to dryness, residue add water 10 ml makes dissolving, adds hydrochloric acid 0.5～2ml again, boiling water bath backflow 1-2 hour, and backflow divides 2-3 extraction with chloroform, each 20ml.Combined chloroform liquid, water bath method, residue add ethanol 2ml makes dissolving, as need testing solution.Other gets Rhizoma Dioscoreae control medicinal material 2g and shines medical material solution in pairs with legal system.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2000), draw above-mentioned two kinds of each 4ul of solution, put respectively on same silica gel g thin-layer plate, with 7-9: 1-3 benzene-acetone, the ammonia saturated with vapor is that developing solvent launches, take out, dry, spray is with the 5-10% ethanol solution of sulfuric acid, and 100-110 ℃ of heating 3-8min is clear to the speckle colour developing.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.
E. get and differentiate that test sample liquid is as test sample liquid under the d item, other gets the Sarsasapogenin reference substance, adds benzene and makes the solution that every 1ml contains 0.5mg, in contrast product solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw above-mentioned two kinds of each 4ul of solution, put respectively on same silica gel g thin-layer plate, with 7-9: 1-3 benzene-acetone, the ammonia saturated with vapor is that developing solvent launches, take out, dry, spray is with 10% sulphuric acid ethanol liquid of 5% vanillin, and 100-110 ℃ of heating 5min is clear to the speckle colour developing.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
Assay: measure according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D), chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; 19-17: 79-82: 1-2 methanol-water-glacial acetic acid is a mobile phase, and the detection wavelength is 250nm.Theoretical cam curve is calculated by puerarin peak should be not less than 4000; The preparation of reference substance solution, precision take by weighing puerarin reference substance 10mg, put in the 25ml measuring bottle, add 30% dissolve with ethanol and are diluted to scale, shake up, precision is measured 2ml, puts in the 10ml measuring bottle, adds 20-40% ethanol to scale, shake up, that is, every 1ml contains puerarin 80 μ g; Get this product granule porphyrize, take by weighing 1.5g, accurate claim surely, put in the tool plug conical flask, the accurate methanol 25ml that adds claims decide weight, and supersound extraction 30-50 minute, put coldly, supply the weight that subtracts mistake with methanol, shake up, filtration is got subsequent filtrate, promptly; Algoscopy, accurate respectively reference substance and each 10 μ l injection chromatograph of liquid of need testing solution drawn measured, promptly; The every 1g of this product contains puerarin (C
21H
20O
9), must not be less than 0.85mg.
Pharmaceutical composition of the present invention (Blood-Sugar lowering particles containing astragalus root and pueraria root etc.) pharmacodynamic study shows: Blood-Sugar lowering particles containing astragalus root and pueraria root etc. has nourishing YIN and clearing away heat, supplementing QI and nourishing YIN, blood circulation promoting and blood stasis dispelling; Be applicable to NIDDM diabetes (deficiency of both QI and YIN hold concurrently syndrome of blood stasis).(1) Blood-Sugar lowering particles containing astragalus root and pueraria root etc. has tangible reduction effect to the blood sugar increasing of ALX-DM rabbit, ALX-DM rabbit carbohydrate metabolism is improved significantly, and the tangible blood sugar increasing that causes of antagonism glucose, and can obviously reduce area under the AUC blood glucose district line.(2) Blood-Sugar lowering particles containing astragalus root and pueraria root etc. has tangible reduction effect to weight loss and the blood sugar increasing of STZ-DM rat, STZ-DM rat carbohydrate metabolism is improved significantly, the blood sugar increasing that glucose is caused has tangible antagonism, and can obviously reduce area under the AUC blood glucose district line.(3) Blood-Sugar lowering particles containing astragalus root and pueraria root etc. improves significantly to the blood lipid dysbolism of STZ-DM rat, shows the reduction of blood CHO, TG level.(4) Blood-Sugar lowering particles containing astragalus root and pueraria root etc. has tangible reduction effect to the blood MDA level of STZ-DM rat, shows that this product has certain antioxidation.(5) Blood-Sugar lowering particles containing astragalus root and pueraria root etc. has certain repair to the islet cells damage that STZ caused, show to the forward and backward serum insulin levels of glucose load that apparently higher than model control group histopathologic examination confirms: Blood-Sugar lowering particles containing astragalus root and pueraria root etc. has certain repair to rat Langerhans islet beta cell damage due to the STZ.(6) Blood-Sugar lowering particles containing astragalus root and pueraria root etc. improves significantly, and can significantly reduce fasting blood glucose level body weight, the endurance decline that alloxan causes diabetic mice; Dynamically 21 days relatively more remarkable (7) Blood-Sugar lowering particles containing astragalus root and pueraria root etc.s of blood sugar reducing function improve significantly to the auricular microcirculation obstacle that alloxan causes diabetic mice behind the blood sugar monitoring prompting medicine.(8) Blood-Sugar lowering particles containing astragalus root and pueraria root etc. does not also have remarkable influence to blood glucose, the blood lipid metabolism of normal rat.
Following experimental example is used to further specify the present invention.
Experimental example one,Blood-Sugar lowering particles containing astragalus root and pueraria root etc. is to the glycometabolic influence of alloxan rabbit
Get 30 of rabbit, the stable raising week taken a sample test fasting glucose twice therebetween, is selected animal standard with 3.6-5.4mmol/L.Press document [1] method: give the injection of alloxan 120mg/kg auricular vein, 48-72h repetition measurement fasting glucose after the modeling, with＞15.0mmol/L as model success animal.
According to the blood sugar increasing amplitude, be divided into water matched group, the high, medium and low dosage group of medicine at random, the insoral group.Press 2.8g/kg, 1.4g/kg, 0.7g/kg, 40mg/kg administration respectively, the administration volume is the 5.0ml/kg body weight, matched group irritates stomach for equal-volume water, successive administration 15 days, fasting 6h before the test, 12h behind the last medicine, ear edge vein exploitating blood do oral glucose tolerance test [2] (the 2.0g/kg glucose is irritated stomach), and, calculate on an empty stomach and glucose is irritated behind the stomach area (Area Under Curve) under 60 minutes blood glucose district lines respectively in 0,30,60,120 minutes repetition measurement blood sugar levels of oral glucose.The result organizes a statistical procedures, sees Table 1, table 2.
AUC=0.5 * 30 minute * [fasting glucose+30 minute blood glucose+30 fens blood glucose+60 minute blood glucose values]
The influence of table 1 pair alloxan model rabbit fasting glucose (mmol/L) (X ± S, n=6)
Blood glucose after the blood glucose administration after the preceding blood glucose modeling of group test
Water group 4.12 ± 0.23 19.45 ± 1.68 17.93 ± 1.82
High dose group 4.20 ± 0.32 18.47 ± 2.26 9.35 ± 1.34***
Middle dosage group 4.25 ± 0.29 19.57 ± 2.07 11.02 ± 2.11***﹠amp;
Low dose group 4.17 ± 0.31 18.38 ± 2.20 11.35 ± 2.60***﹠amp;
Insoral group 4.25 ± 0.34 19.43 ± 1.96 8.63 ± 1.35***
TANGMAIKANG group 4.35 ± 0.34 19.52 ± 2.27 11.08 ± 3.06***
Annotate: compare before testing with self: ### shows P＜0.001.
Compare with the modeling matched group behind the medicine: * shows P＜0.05, and * * * shows P＜0.001.
Table 1 is the result show: Blood-Sugar lowering particles containing astragalus root and pueraria root etc. has tangible reduction effect to the blood sugar increasing of alloxan model rabbit, and high, middle dosage group and model control group more all have utmost point significant difference, and low dose group and model control group more all have significant difference; Blood-Sugar lowering particles containing astragalus root and pueraria root etc. improves significantly to alloxan model rabbit carbohydrate metabolism, and the blood sugar increasing that glucose is caused has tangible reduction effect, and high, medium and low dosage group and model control group more all have significant difference.
Experimental example two,The antioxidation of Blood-Sugar lowering particles containing astragalus root and pueraria root etc.---to the influence of STZ-DM rat serum MDA content:
Get 70 of rats, the stable raising week taken a sample test fasting glucose twice therebetween, is selected animal standard with 3.6-5.4mmol/L.Wherein 10 rats are as the normal control group, other 60 rats are pressed document [3] method: with 0.1mol/LpH4.5 sodium citrate-citrate buffer solution, the STZ solution of fresh configuration 10mg/ml in the ice territory, every rat is pressed the 50mg/kg intravenous injection, 7-10 days repetition measurement fasting glucose after the modeling, with＞13.8mmol/L as model success rat.60 of STZ-DM rats are divided into water matched group, the high, medium and low dosage group of medicine, insoral group, TANGMAIKANG group at random by the blood sugar increasing amplitude.Press 3.24g/kg, 1.62g/kg, 0.81g/kg, 0.009g/kg, 1.35g/kg administration respectively, the administration volume is the 1.0ml/100g body weight, and matched group irritates stomach, successive administration 20 days for equal-volume water; The socket of the eye venous blood collection is pressed the TCA-TBA method, and in wavelength 535nm test sample trap A value, with reflection blood MDA level, the result organizes a statistical procedures, sees Table 2
Table 2. Blood-Sugar lowering particles containing astragalus root and pueraria root etc. to the influence of STZ-DM rat serum MDA content (X ± s, n=10)
Group blood MDA content
Normal control group 0.071 ± 0.023
Model control group 0.148 ± 0.024###
High dose group 0.112 ± 0.027##**
Middle dosage group 0.118 ± 0.027####*
Low dose group 0.124 ± 0.022###*
Insoral group 0.121 ± 0.033###
TANGMAIKANG group 0.116 ± 0.026###*
Table 2 is the result show: MDA organizes in the peroxidating process very important intermediate product, and Blood-Sugar lowering particles containing astragalus root and pueraria root etc. has tangible reduction effect to the blood MDA level of STZ-DM rat, and high, medium and low dosage group and model control group more all have significant difference.Show that this product has certain antioxidation.
Experimental example three,Blood-Sugar lowering particles containing astragalus root and pueraria root etc. is to the influence of STZ-DM rat Langerhans islet beta cell function
Experimental animal grouping, modeling, administration time and dosage is all with experimental example 2, fasting 8h before the test, and 12h behind the last medicine is doing oral glucose tolerance test simultaneously, and oral glucose 0,30,60,120 minutes, the eye socket venous blood collection was surveyed insulin level respectively.The result organizes a statistical procedures, sees Table 3.
The influence of table 3. pair STZ rat model serum insulin levels (X ± S, n=8)
Group gives behind the glucose 0 to give behind the glucose 30 and give behind the glucose 60 and give behind the glucose 120 fens
Normal group 4.99 ± 1.18 4.81 ± 1.09 5.06: 0.59 3.26: 1.86
Model group 1.49 ± 0.93### 1.26 ± 0.33 1.30: 0.40### 1.47: 0.51
High dose 3.19 ± 2.02#* 3.75 ± 1.33*** 2.93: 1.26###** 2.82: 0.71***
Middle dosage 3.76 ± 1.72**﹠amp; 3.03 ± 1.79#* 2.06: 0.42###** 2.19: 0.72*
Low dosage 1.97 ± 0.54### 2.07 ± 0.37###***$$﹠amp; 2.13: 1.01 ###* 1.69: 0.86#$﹠amp;
Insoral 2.23 ± 0.78### 3.51 ± 1.83** 2.49: 0.85###** 2.78: 0.69***
TANGMAIKANG 2.68 ± 0.71###* 2.83 ± 1.53##* 1.57: 0.76###$﹠amp; 1.96: 0.60$﹠amp;
Table 3 is the result show: the model control group blood insulin levels is starkly lower than the normal control group before giving glucose load, illustrates that STZ causes certain damage to beta Cell of islet.Medicine height, middle dosage group insulin level illustrate that apparently higher than model control group medicine has certain repair to the β cell injury that STZ causes.30 minutes insulin secretions increased after normal rat was given glucose load, the insulin secretion peak occurred, and descended gradually thereafter in 60 minutes.Medicine height, middle dosage group 30 minutes serum insulin levels after giving glucose load raise rapidly, and apparently higher than the modeling matched group, have certain effect for the carbohydrate metabolism that improves the STZ-DM rat.
Embodiment 1:
Radix Astragali 600g Radix Puerariae 400g Rhizoma Dioscoreae 400g Rhizoma Atractylodis 300g
Rhizoma Anemarrhenae 300g Radix Trichosanthis 300g Radix Rehmanniae 400g Herba Portulacae 300g
Radix Salviae Miltiorrhizae 300g Ramulus Euonymi 300g Ramulus Mori 300g
More than ten simply, get Rhizoma Atractylodis, Radix Salviae Miltiorrhizae two flavors with 6 times of amount 95% alcohol reflux 1 hour, filtration, medicinal residues and Radix Puerariae merge, with 70%6 alcohol reflux three times, and each 5 times of amounts, extracted respectively 2 hours, 1 hour, 0.5 hour, filter, merging filtrate, decompression recycling ethanol (60 ~ 70 ℃ ,-0.08Mpa), water liquid is standby; Get eight flavor medicines such as the Radix Astragali and decoct with water secondary, add 10 times of decoctings for the first time and boiled 1.5 hours, add 8 times of decoctings for the second time and boiled 1.5 hours, collecting decoction filters, and filtrate is concentrated into relative density 1.10 ~ 1.12 (60 ℃), be blended into above-mentioned alcohol extraction concentrated solution, add an amount of dextrin (about 150g) and stir evenly, spray drying (180 ~ 200 ℃ of air intakes, 80 ~ 85 ℃ of air-out), dry powder adds protein sugar 5g, and adds dextrin to 1000g, granule is made in dry-pressing, and every packed 5g is oral, one time 1 bag, 3 times on the one.
Embodiment 2:The method of quality control of medicament composition granule agent of the present invention:
Differentiate that a. gets this product powder 2g, add methanol 40ml supersound extraction 30 minutes, filter, filtrate evaporate to dryness, residue add water 15ml makes dissolving, 30ml, 30ml, 15ml, 15ml divides four extractions with ether, discards ether solution, 30ml, 30ml, 15ml water intaking layer divides three extractions with water-saturated n-butanol, merges n-butanol layer and washs at twice with 1% sodium hydroxide, each 15ml, discard alkali liquor, n-butyl alcohol with n-butyl alcohol saturated be washed to neutrality, get n-butyl alcohol liquid, be concentrated into dried, residue adds water 5ml makes dissolving, is added on internal diameter 1.5cm, the D that high 10cm has handled well
101Macroporous resin column, difference water 50ml, 40% ethanol 30ml, 70% ethanol 50ml eluting is collected 70% ethanol elution, and evaporate to dryness, residue add methanol 2ml makes dissolving, as need testing solution.Other gets the astragaloside reference substance, adds methanol and makes solution that every 1ml contains 1mg product solution in contrast.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B); Draw need testing solution 8 μ l, astragaloside reference substance 4 μ l, put respectively on same silica gel g thin-layer plate, lower floor's solution of placing below 10 ℃ with 13: 7: 2 chloroform-methanol-water is developing solvent, launches, and takes out, dry, spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ are heated 5min, and is clear to the speckle colour developing.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
B. get this product 3g, add ethanol 30ml, supersound extraction 30 minutes filters, and filtrate is concentrated into 2ml, as need testing solution.Other gets Rhizoma Atractylodis control medicinal material 2g, shines medical material solution in pairs with legal system, according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B) test, draw need testing solution 4 μ l, control medicinal material solution 2 μ l put respectively on same silica gel G plate, with 60-90 ℃ of petroleum ether is developing solvent, launches, and takes out, dry, spray is with 10% ethanol solution of sulfuric acid of 5% vanillin, and it is clear that hot blast blows to the speckle colour developing, in the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the speckle of same color.
C. get and differentiate that need testing solution is as need testing solution under the b item, other gets Radix Salviae Miltiorrhizae control medicinal material 2g, adds ethanol 30ml, and supersound extraction 30 minutes filters, and filtrate is concentrated into 2ml, in contrast medical material solution.Get the tanshinone reference substance again, add ethyl acetate and make the solution that every 1ml contains 2mg.According to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2000) test, draw each 4 μ l of above-mentioned three kinds of solution, reference substance solution 2 μ l put respectively on same silica gel g thin-layer plate, are developing solvent with 19: 1 benzene-ethyl acetates, launch, and take out, and dry.In the test sample chromatograph, with control medicinal material chromatograph and reference substance chromatograph relevant position on, show the speckle of same color.
D. get this product 2g, use ultrasonic 30 minutes of methanol 20ml, filter, filtrate evaporate to dryness, residue add water 10ml makes dissolving, adds hydrochloric acid 1ml again, and boiling water bath refluxed 1 hour, and backflow extracts at twice with chloroform, each 20ml.Combined chloroform liquid, water bath method, residue add ethanol 2ml makes dissolving, as need testing solution.Other gets Rhizoma Dioscoreae control medicinal material 2g and shines medical material solution in pairs with legal system.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2000), draw above-mentioned two kinds of each 4ul of solution, put respectively on same silica gel g thin-layer plate, with 9.1 benzene-acetone, the ammonia saturated with vapor is for developing solvent launches, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ of heating 5min are clear to the speckle colour developing.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.
E. get and differentiate that test sample liquid is as test sample liquid under the d item, other gets the Sarsasapogenin reference substance, adds benzene and makes the solution that every 1ml contains 0.5mg, in contrast product solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw above-mentioned two kinds of each 4ul of solution, put respectively on same silica gel g thin-layer plate, with 9: 1 benzene-acetone, the ammonia saturated with vapor was for developing solvent launches, take out, dry, spray is with 10% sulphuric acid ethanol liquid of 5% vanillin, and 105 ℃ of heating 5min are clear to the speckle colour developing.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
Assay: measure according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D), chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; 19: 81: 1 methanol-water-glacial acetic acid are mobile phase, and the detection wavelength is 250nm.Theoretical cam curve is calculated by puerarin peak should be not less than 4000; The preparation of reference substance solution, precision take by weighing puerarin reference substance 10mg, put in the 25ml measuring bottle, add 30% dissolve with ethanol and are diluted to scale, shake up, and precision is measured 2ml, puts in the 10ml measuring bottle, adds 30% ethanol to scale, shake up, that is, every 1ml contains puerarin 80 μ g; Get this product granule porphyrize, take by weighing 1.5g, accurate claim surely, put in the tool plug conical flask, the accurate methanol 25ml that adds claims decide weight, 250w, and 50kHz supersound extraction 40 minutes is put coldly, supplies the weight that subtracts mistake with methanol, shakes up, and filtration is got subsequent filtrate, promptly; Algoscopy, accurate respectively reference substance and each 10 μ l injection chromatograph of liquid of need testing solution drawn measured, promptly; The every 1g of this product contains puerarin (C
21H
20O
9), must not be less than 0.85mg.
Claims (8)
1. pharmaceutical composition for the treatment of diabetes is characterized in that this pharmaceutical composition made by following crude drug:
Radix Astragali 400-800 weight portion Radix Puerariae 300-500 weight portion
Rhizoma Dioscoreae 300-500 weight portion Rhizoma Atractylodis 200-400 weight portion
Rhizoma Anemarrhenae 200-400 weight portion Radix Trichosanthis 200-400 weight portion
Radix Rehmanniae 300-500 weight portion Herba Portulacae 200-400 weight portion
Radix Salviae Miltiorrhizae 200-400 weight portion Ramulus Euonymi 200-400 weight portion
Ramulus Mori 200-400 weight portion.
2. a kind of pharmaceutical composition for the treatment of diabetes as claimed in claim 1 is characterized in that this pharmaceutical composition made by following crude drug:
The Radix Astragali 600 weight portion Radix Puerariaes 400 weight portion Rhizoma Dioscoreaes 400 weight portions
The Rhizoma Atractylodis 300 weight portion Rhizoma Anemarrhenaes 300 weight portion Radix Trichosanthis 300 weight portions
Radix Rehmanniae 400 weight portion bitterroots 300 weight portion Radix Salviae Miltiorrhizaes 300 weight portions
Ramulus Euonymi 300 weight portion Ramulus Moris 300 weight portions.
3. a kind of pharmaceutical composition for the treatment of diabetes as claimed in claim 1 or 2 is characterized in that this pharmaceutical composition crude drug can make any clinical acceptable forms, as tablet, and capsule, granule, oral liquid, subcutaneous administration preparation, suppository.
4. a kind of pharmaceutical composition for the treatment of diabetes as claimed in claim 5 is characterized in that this preparation of drug combination method is:
Get Rhizoma Atractylodis, Radix Salviae Miltiorrhizae two flavors and doubly measured the 85-95% alcohol reflux 1-2 hour with 4-8, filter, medicinal residues merge with Radix Puerariae, usefulness 60-80% alcohol reflux 2-4 time, each 4-7 doubly measures, and extracts respectively 0.5-2 hour, filters, merging filtrate, decompression recycling ethanol, water liquid is standby; Get eight flavor medicines such as the Radix Astragali and decoct with water 2-3 time, add 8-12 times of water at every turn, decocted 1-2 hour, collecting decoction filters, and filtrate is concentrated into 1.10 ~ 1.12/60 ℃ of relative densities, be blended into above-mentioned alcohol extraction concentrated solution, spray drying is made clinical acceptable forms, as tablet, capsule, granule, oral liquid, the subcutaneous administration preparation, suppository.
5. a kind of pharmaceutical composition for the treatment of diabetes as claimed in claim 4 is characterized in that this preparation of drug combination method is:
Get Rhizoma Atractylodis, Radix Salviae Miltiorrhizae two flavors with 6 times of amount 95% alcohol reflux 1 hour, filter, medicinal residues and Radix Puerariae merging, with 70% alcohol reflux three times, each 5 times of amounts, extracted respectively 2 hours, and 1 hour, 0.5 hour, filtered, merging filtrate, 60 ~ 70 ℃ ,-0.08Mpa decompression recycling ethanol, water liquid is standby; Get eight flavor medicines such as the Radix Astragali and decoct with water secondary, add 10 times of decoctings for the first time and boiled 1.5 hours, add 8 times of decoctings for the second time and boiled 1.5 hours, collecting decoction filters, and filtrate is concentrated into 1.10 ~ 1.12/60 ℃ of relative densities, be blended into above-mentioned alcohol extraction concentrated solution, add dextrin and stir evenly spray drying, 180 ~ 200 ℃ of air intakes, 80 ~ 85 ℃ of air-out, dry powder adds protein sugar, and adds dextrin to 1000 weight portion, and granule is made in dry-pressing.
6. as the method for quality control of granule as described in the claim 5, it is characterized in that comprising in this method and differentiate and/or assay, wherein differentiate to comprise in the following method one or more:
Differentiate that a. gets this product powder 2g, add methanol 40ml supersound extraction 20-40 minute, filter, filtrate evaporate to dryness, residue add water 15ml makes dissolving, divides 2-4 extraction with ether, each 15-30ml, discard ether solution, the water intaking layer divides 2-4 extraction with water-saturated n-butanol, each 15-30ml, merge n-butanol layer and divide 2-3 washing with 0.5～2% sodium hydroxide solution, each 10-20ml discards alkali liquor, n-butyl alcohol with n-butyl alcohol saturated be washed to neutrality, get n-butyl alcohol liquid, be concentrated into driedly, residue adds water 5ml makes dissolving, is added on the D that has handled well
101Macroporous resin column, difference water 40-60ml, 30-50% ethanol 20-40ml, 60-80% ethanol 40-60ml eluting is collected the 60-80% ethanol elution, and evaporate to dryness, residue add methanol 2ml makes dissolving, as need testing solution; Other gets the astragaloside reference substance, adds methanol and makes solution that every 1ml contains 1mg product solution in contrast.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B); Draw need testing solution 8 μ l, astragaloside reference substance 4 μ l, put respectively on same silica gel g thin-layer plate, with 11-14: 6-9: lower floor's solution that 1-2 chloroform-methanol-water is placed below 10 ℃ is developing solvent, launches, and takes out, dry, spray is with 10% ethanol solution of sulfuric acid, and 100-110 ℃ is heated 5-8min, and is clear to the speckle colour developing.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
B. get this product 3g, add ethanol 30ml, supersound extraction 20-40 minute, filter, filtrate is concentrated into 2ml, as need testing solution.Other gets Rhizoma Atractylodis control medicinal material 2g, shines medical material solution in pairs with legal system, according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B) test, draw need testing solution 4 μ l, control medicinal material solution 2 μ l put respectively on same silica gel G plate, with 60-90 ℃ of petroleum ether is developing solvent, launches, and takes out, dry, spray is with 10% ethanol solution of sulfuric acid of 5% vanillin, and it is clear that hot blast blows to the speckle colour developing, in the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the speckle of same color;
C. get and differentiate that need testing solution is as need testing solution under the b item, other gets Radix Salviae Miltiorrhizae control medicinal material 2g, adds ethanol 30ml, and supersound extraction 20-40 minute, filter, filtrate is concentrated into 2ml, in contrast medical material solution.Get the tanshinone reference substance again, add ethyl acetate and make the solution that every 1ml contains 2mg; According to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2000) test, draw each 4 μ l of above-mentioned three kinds of solution, reference substance solution 2 μ l, put respectively on same silica gel g thin-layer plate, with 16-19: 1-4 benzene-ethyl acetate is developing solvent, launches, take out, dry; In the test sample chromatograph, with control medicinal material chromatograph and reference substance chromatograph relevant position on, show the speckle of same color;
D. get this product 2g, use methanol 20ml ultrasonic 20-40 minute, filter, filtrate evaporate to dryness, residue add water 10ml makes dissolving, adds hydrochloric acid 0.5～2ml again, boiling water bath backflow 1-2 hour, and backflow divides 2-3 extraction with chloroform, each 20ml.Combined chloroform liquid, water bath method, residue add ethanol 2ml makes dissolving, as need testing solution.Other gets Rhizoma Dioscoreae control medicinal material 2g and shines medical material solution in pairs with legal system; Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2000), draw above-mentioned two kinds of each 4ul of solution, put respectively on same silica gel g thin-layer plate, with 7-9: 1-3 benzene-acetone, the ammonia saturated with vapor is that developing solvent launches, take out, dry, spray is with the 5-10% ethanol solution of sulfuric acid, and 100-110 ℃ of heating 3-8min is clear to the speckle colour developing; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color;
E. get and differentiate that test sample liquid is as test sample liquid under the d item, other gets the Sarsasapogenin reference substance, adds benzene and makes the solution that every 1ml contains 0.5mg, in contrast product solution; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw above-mentioned two kinds of each 4ul of solution, put respectively on same silica gel g thin-layer plate, with 7-9: 1-3 benzene-acetone, the ammonia saturated with vapor is that developing solvent launches, take out, dry, spray is with 10% sulphuric acid ethanol liquid of 5% vanillin, and 100-110 ℃ of heating 5min is clear to the speckle colour developing; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
Assay: measure according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D), chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; 19-17: 79-82: 1-2 methanol-water-glacial acetic acid is a mobile phase, and the detection wavelength is 250nm; Theoretical cam curve is calculated by puerarin peak should be not less than 4000; The preparation of reference substance solution, precision take by weighing puerarin reference substance 10mg, put in the 25ml measuring bottle, add 30% dissolve with ethanol and are diluted to scale, shake up, precision is measured 2ml, puts in the 10ml measuring bottle, adds 20-40% ethanol to scale, shake up, that is, every 1ml contains puerarin 80 μ g; Get this product granule porphyrize, take by weighing 1.5g, accurate claim surely, put in the tool plug conical flask, the accurate methanol 25ml that adds claims decide weight, and supersound extraction 30-50 minute, put coldly, supply the weight that subtracts mistake with methanol, shake up, filtration is got subsequent filtrate, promptly; Algoscopy, accurate respectively reference substance and each 10 μ l injection chromatograph of liquid of need testing solution drawn measured, promptly; The every 1g of this product contains puerarin (C
21H
20O
9), must not be less than 0.85mg.
7, as the method for quality control of granule as described in the claim 6, it is characterized in that this method is: differentiate
A. get this product powder 2g, add methanol 40ml supersound extraction 30 minutes, filter, filtrate evaporate to dryness, residue add water 15ml makes dissolving, 30ml, 30ml, 15ml, 15ml divides four extractions with ether, discards ether solution, 30ml, 30ml, 15ml water intaking layer divides three extractions with water-saturated n-butanol, merges n-butanol layer and washs at twice with 1% sodium hydroxide, each 15ml, discard alkali liquor, n-butyl alcohol with n-butyl alcohol saturated be washed to neutrality, get n-butyl alcohol liquid, be concentrated into dried, residue adds water 5ml makes dissolving, is added on internal diameter 1.5cm, the D that high 10cm has handled well
101Macroporous resin column, difference water 50ml, 40% ethanol 30ml, 70% ethanol 50ml eluting is collected 70% ethanol elution, and evaporate to dryness, residue add methanol 2ml makes dissolving, as need testing solution.Other gets the astragaloside reference substance, adds methanol and makes solution that every 1ml contains 1mg product solution in contrast.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B); Draw need testing solution 8 μ l, astragaloside reference substance 4 μ l, respectively on the same silica gel g thin-layer plate of idea, lower floor's solution of placing below 10 ℃ with 13: 7: 2 chloroform-methanol-water is developing solvent, launches, and takes out, dry, spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ are heated 5min, and is clear to the speckle colour developing.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
B. get this product 3g, add ethanol 30ml, supersound extraction 30 minutes filters, and filtrate is concentrated into 2ml, as need testing solution.Other gets Rhizoma Atractylodis control medicinal material 2g, shines medical material solution in pairs with legal system, according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B) test, draw need testing solution 4 μ l, control medicinal material solution 2 μ l put respectively on same silica gel G plate, with 60-90 ℃ of petroleum ether is developing solvent, launches, and takes out, dry, spray is with 10% ethanol solution of sulfuric acid of 5% vanillin, and it is clear that hot blast blows to the speckle colour developing, in the test sample chromatograph, with control medicinal material chromatograph relevant position on, show the speckle of same color.
C. get and differentiate that need testing solution is as need testing solution under the b item, other gets Radix Salviae Miltiorrhizae control medicinal material 2g, adds ethanol 30ml, and supersound extraction 30 minutes filters, and filtrate is concentrated into 2ml, in contrast medical material solution.Get the tanshinone reference substance again, add ethyl acetate and make the solution that every 1ml contains 2mg.According to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2000) test, draw each 4 μ l of above-mentioned three kinds of solution, reference substance solution 2 μ l put respectively on same silica gel g thin-layer plate, are developing solvent with 19: 1 benzene-ethyl acetates, launch, and take out, and dry.In the test sample chromatograph, with control medicinal material chromatograph and reference substance chromatograph relevant position on, show the speckle of same color.
D. get this product 2g, use ultrasonic 30 minutes of methanol 20ml, filter, filtrate evaporate to dryness, residue add water 10ml makes dissolving, adds hydrochloric acid 1ml again, and boiling water bath refluxed 1 hour, and backflow extracts at twice with chloroform, each 20ml.Combined chloroform liquid, water bath method, residue add ethanol 2ml makes dissolving, as need testing solution.Other gets Rhizoma Dioscoreae control medicinal material 2g and shines medical material solution in pairs with legal system.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2000), draw above-mentioned two kinds of each 4ul of solution, put respectively on same silica gel g thin-layer plate, with 9: 1 benzene-acetone, the ammonia saturated with vapor was for developing solvent launches, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and 105 ℃ of heating 5min are clear to the speckle colour developing.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.
E. get and differentiate that test sample liquid is as test sample liquid under the d item, other gets the Sarsasapogenin reference substance, adds benzene and makes the solution that every 1ml contains 0.5mg, in contrast product solution.Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 B), draw above-mentioned two kinds of each 4ul of solution, put respectively on same silica gel g thin-layer plate, with 9: 1 benzene-acetone, the ammonia saturated with vapor was for developing solvent launches, take out, dry, spray is with 10% sulphuric acid ethanol liquid of 5% vanillin, and 105 ℃ of heating 5min are clear to the speckle colour developing.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
Assay: measure according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2000 D), chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; 19: 81: 1 methanol-water-glacial acetic acid are mobile phase, and the detection wavelength is 250nm.Theoretical cam curve is calculated by puerarin peak should be not less than 4000;
The preparation of reference substance solution, precision take by weighing puerarin reference substance 10mg, put in the 25ml measuring bottle, add 30% dissolve with ethanol and are diluted to scale, shake up, and precision is measured 2ml, puts in the 10ml measuring bottle, adds 30% ethanol to scale, shake up, that is, every 1ml contains puerarin 80 μ g; Get this product granule porphyrize, take by weighing 1.5g, accurate claim surely, put in the tool plug conical flask, the accurate methanol 25ml that adds claims decide weight, 250w, and 50kHz supersound extraction 40 minutes is put coldly, supplies the weight that subtracts mistake with methanol, shakes up, and filtration is got subsequent filtrate, promptly;
Algoscopy, accurate respectively reference substance and each 10 μ l injection chromatograph of liquid of need testing solution drawn measured, promptly; The every 1g of this product contains puerarin (C
21H
20O
9), must not be less than 0.85mg.
10, the application of pharmaceutical composition as claimed in claim 1 or 2 in the medicine of preparation treatment diabetes.
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| Application Number | Priority Date | Filing Date | Title |
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| CNA021594627A CN1511576A (en) | 2002-12-31 | 2002-12-31 | Medicinal composition for preventing and curing diabetes and its preparing method |
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| CNA021594627A CN1511576A (en) | 2002-12-31 | 2002-12-31 | Medicinal composition for preventing and curing diabetes and its preparing method |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1321648C (en) * | 2005-06-30 | 2007-06-20 | 华中科技大学同济医学院附属同济医院 | Insulin sensitive-intensifying agent from plant and its preparing method and use |
| CN101406598B (en) * | 2008-11-13 | 2011-02-16 | 刘严明 | Chinese medicine composite for treating diabetes and preparation method thereof |
| CN103211996A (en) * | 2013-05-06 | 2013-07-24 | 武汉诺贝药业有限公司 | Preparation method of traditional Chinese medicine for treating diabetes mellitus |
| CN104491416A (en) * | 2015-01-20 | 2015-04-08 | 刘壮志 | Traditional Chinese medicine composition used for treating diabetes |
-
2002
- 2002-12-31 CN CNA021594627A patent/CN1511576A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1321648C (en) * | 2005-06-30 | 2007-06-20 | 华中科技大学同济医学院附属同济医院 | Insulin sensitive-intensifying agent from plant and its preparing method and use |
| CN101406598B (en) * | 2008-11-13 | 2011-02-16 | 刘严明 | Chinese medicine composite for treating diabetes and preparation method thereof |
| CN103211996A (en) * | 2013-05-06 | 2013-07-24 | 武汉诺贝药业有限公司 | Preparation method of traditional Chinese medicine for treating diabetes mellitus |
| CN104491416A (en) * | 2015-01-20 | 2015-04-08 | 刘壮志 | Traditional Chinese medicine composition used for treating diabetes |
| CN104491416B (en) * | 2015-01-20 | 2017-12-15 | 刘准 | A kind of Chinese medicine composition for being used to treat diabetes |
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