CN101347495A - Preparation of Glabridin dispersible tablets and use of the tablets in reducing blood sugar as medicament active composition - Google Patents
Preparation of Glabridin dispersible tablets and use of the tablets in reducing blood sugar as medicament active composition Download PDFInfo
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Abstract
The present invention discloses a preparation process of glycyrrhiza flavonoid dispersible tablet and the application of the tablet as the active component in decreasing blood sugar. The activity test proves three dosage groups of the glycyrrhiza flavonoid dispersible tablet all play a significant role in decreasing blood sugar of hyperglycemic model mice caused by alloxan, adrenalin and dextrose; the role in decreasing blood sugar is equal to or more significant than similar sugar-reducing medicines in clinical use. At the same time, the 'three more and one less' symptom of the diabetic model mouse is obviously relieved and especially the drinking amount is obviously reduced. The mouse acute toxicity test proves that under the maximum dosage, the mouse has no toxic side effect and the weight growth is normal. The source of raw materials is wide, the main sugar-reducing component is taken from natural medicines and the sugar-reducing effect is obvious; therefore, the present invention is suitable for long-term use. The present invention with simple preparation technology and low production cost is suitable for large-scale industrial production.
Description
Technical field
The present invention relates to isolation and purification method and the preparation of licoflavone dispersible tablet and the experimental study of licoflavone dispersible tablet hypoglycemic activity of the effective sugar-lowering components of licoflavone dispersible tablet.
Background technology
Diabetes be a kind of because of insulin secretion absolute or relative deficiency, causing with the hyperglycemia is the endocrine metabolism disease of feature.If body is in hyperglycemia state for a long time, can cause losing one's sight, multiple complications such as renal failure, neuropathy, cardiovascular and cerebrovascular disease, extremity gangrene.Now become the third-largest disease of the serious harm human health after cardiovascular and cerebrovascular vessel, cancer.For the treatment of type i diabetes, research direction is exploitation convenient drug administration, effective insulin preparation and succedaneum at present.And for the treatment of type ii diabetes, then mainly be to rely on existing orally-taken blood sugar reducing medicine to carry out limited treatment.
Alpha-glucosidase inhibitor is a class new oral hypoglycemic drug of developing the seventies in last century, can the competitive inhibition small intestinal in the activity of alpha-glucosidase, delay or suppress glucose in intestinal absorption, effectively reduce postprandial hyperglycemia.Succeeding in developing of such medicine is for the diabetes oral hypoglycemic drug therapy has been opened up a new approach.Alpha-glucosidase inhibitor is mainly used in treatment of diabetes clinically, can use the treatment type ii diabetes separately or with sulphanylureas, biguanides and insulin combination, also can in the treatment of type i diabetes, share with insulin, reduce the consumption and the stable blood glucose of insulin, and alleviate hyperinsulinemia.
At present, clinical alpha-glucosidase inhibitor commonly used has acarbose (acarbose), voglibose (voglibose), miglitol (miglitol) etc.But these medicines not only cost an arm and a leg, and find in the clinical practice that bigger toxic and side effects is arranged, can not life-time service.As recent discovery acarbose (acarbose) side effect that damages liver is arranged, might cause serious hepatic insufficiency (" Chinese diabetes magazine " 2002,10 (6): 326-329).Be badly in need of the developmental research natural origin, toxic and side effects is little, but the alpha-glucosidase inhibitor of life-time service.
The Chinese medicine Radix Glycyrrhizae is glycyrrhizic legume (Glycyrrhiza uralensis Fisch.), the dry root and rhizome of Glycyrrhiza inflata Bat. (Glycyrrhiza inflata Bat.) or Glycyrrhiza glabra L. (Glycyrrhiza glabra L.).China is familiar with in the world and studies Radix Glycyrrhizae country the earliest, and is promptly on the books in the Shennong's Herbal of the Eastern Han Dynasty.Its sweet in the mouth, property is flat, goes into the heart, lung, spleen, stomach warp.Disease such as be used for the treatment of in weakness of the spleen and stomach, deficiency of QI in middle-JIAO, cough and asthma, skin sore, the abdomen contraction and have a pain and relax medicine strong, separate poison of drug.Scientific research experiment shows: be rich in compositions such as glycyrrhizic acid, enoxolone, flavone, saponin, alkaloid and aminoacid in the Radix Glycyrrhizae, effect such as have antiviral, antibiotic, antiulcer, antioxidation, antitumor, detoxify, protect the liver.In recent years, along with further investigation, show that Radix Glycyrrhizae has effects such as antiviral, antibiotic, antiulcer, antiinflammatory, blood fat reducing, analgesia to the Radix Glycyrrhizae pharmacological action.
Summary of the invention
Radix Glycyrrhizae is as a kind of conventional Chinese medicine and spice, and aboundresources is cheap, uses very extensively, has the value and the research background of further development and use.Extraction separation goes out the blood sugar lowering effective ingredient from the Chinese medicine Radix Glycyrrhizae through studying for many years in our seminar.Purpose of the present invention aims to provide a kind of licoflavone dispersible tablet preparation method.
In addition then, in test, the licoflavone dispersible tablet is made medicament active composition and is manifested day by day in pharmacology's effect, and the diabetic mice experimental study shows that the said preparation hypoglycemic effect is obvious, and another object of the present invention is intended to above-mentioned tablet and does the application of medicament active composition in blood sugar lowering.
Purpose of the present invention can be achieved through the following technical solutions:
The preparation of licoflavone dispersible tablet, its step is the extracting liquorice decoction pieces, in 1: 4 ratio reflux, extract, of Radix Glycyrrhizae and 95% ethanol 3 times, each 2h, merging filtrate, reclaim solvent, get extractum, gained extractum is passed through 30-60 order polyamide column chromatography, water, 95% ethanol elution successively, with hydrochloric acid-magnesium powder reaction detection merge total flavones, again through concentrating and lyophilization gets the total flavones powder; Get 4-12% carboxymethyl starch sodium (CMS-Na), 2-6%L-low substituted hydroxy-propyl fiber (HPC), 0.5-1.5% sodium lauryl sulphate (SLS) and 20-85% licoflavone powder, add an amount of microcrystalline Cellulose (MCC), cross 100 mesh sieve mix homogeneously, after 60 ℃ of dryings, direct compression.
Another object of the present invention is all to have significant hypoglycemic activity to realize by the hyperglycemia model mice that three dosage groups of licoflavone dispersible tablet cause alloxan, epinephrine, glucose.
The beneficial effect of advantage of the present invention and generation:
1, preparation of the present invention, main sugar-lowering components is taken from natural drug, the crude drug wide material sources, preparation technology is simple, and the required cost of production is lower, suitability for mass industrialized production.
2, preparation of the present invention, hypoglycemic effect is obvious, and toxic and side effects is low, is suitable for life-time service.The hyperglycemia mouse model that preparation of the present invention causes alloxan, epinephrine and glucose all has significant hypoglycemic activity (see Table 4, table 5, table 6).The similar hypoglycemic medicine acarbose with clinical use of its blood sugar reducing function quite or more remarkable." three-many-one-little " symptom of diabetic mice is obviously alleviated simultaneously, and especially amount of drinking water obviously reduces.With the serious liver injury of acarbose with might cause that serious side effect such as hepatic insufficiency compare, this preparation acute toxicity test in mice studies show that said preparation toxicity is very low, does not see toxicity under the maximum dosage, weight of mice is normal.
3, the every quality of preparation of the present invention all meets the prescription of pharmacopeia dispersible tablet.Is to investigate index with the disintegration time, the prescription of licoflavone dispersible tablet is formed and preparation technology has carried out primary election and orthogonal test preferably (seeing Table 1 factor level table).The optimum prescription of determining the licoflavone dispersible tablet is 4%CMS-N
a, 4%L-HPC, 1.5%SLS, 25% licoflavone and 65.5%MCC, dry method direct compression (250mg/ sheet) (sees Table 2L
9(3
3) test card).Dispersible tablet disintegration time, dispersing uniformity and the dissolution (seeing Fig. 1 licoflavone dispersible tablet dissolution curve chart) that makes by this technology all meets the prescription of pharmacopeia dispersible tablet.
4, preparation stability of the present invention is good.The present invention studies the preparation stability of licoflavone dispersible tablet, and the licoflavone dispersible tablet is at 40 ℃, and illumination is 4000Lx, and relative humidity is to place 3 months under 75% condition, and the content of licoflavone does not have significant change (the results are shown in Table 3).
Description of drawings
Fig. 1 licoflavone dispersible tablet dissolution curve chart
The specific embodiment
(1) preparation of licoflavone dispersible tablet
Embodiment 1
Take by weighing licorice piece 2000g, in 1: 4 ratio reflux, extract, of Radix Glycyrrhizae and 95% ethanol 3 times, each 2h, merge 3 times filtrate, vacuum distillation recovered solvent gets extractum 356g, gained extractum is passed through 30-60 order polyamide column chromatography, water, 95% ethanol elution successively, with hydrochloric acid-magnesium powder reaction detection, merge total flavones.Get total flavones powder 82.45g through concentrating with lyophilization again, productive rate is 4.12%.Take by weighing 4%CMS-Na, 4%L-HPC, 1.5%SLS and 20% flavone powder by recipe quantity, add an amount of MCC, cross 100 mesh sieve mix homogeneously, after 60 ℃ of dryings, direct compression (every contains effective composition 250mg).
The preparation technology of licoflavone dispersible tablet (LFT)
(1) preparation technology's is preferred
According to document and trial test, the principal element that influences dispersible tablet preparation technology has disintegrating agent, binding agent, surfactant, pressure etc.But because licoflavone itself has certain moisture and adhesive, thus this test with disintegration time as index, only investigate the consumption of disintegrating agent carboxymethyl base Starch Sodium and low-substituted hydroxypropyl cellulose, surfactant sodium lauryl sulphate.Test arrangement sees Table 1.
The factor level table of table 1. licoflavone dispersible tablet preparation
Tab.1?The?table?of?factor?and?level
By the dispersible tablet that the experimental establishment of table 1 makes, check by dispersible tablet inspection technique disintegration of pharmacopeia regulation respectively, promptly in (20 ± 1) ℃ water, the whole disintegrates of 3min planted agent.
Result of the test (seeing Table 2)
Table 2 L
9(3
3) test card
Tab.2?The?table?of?L
9(3
3)test
Through intuitive analysis, each factor is B＞C＞A to the size order that influences of disintegration time as can be known, and definite optimised process prescription is for the consumption of CMS-Na is 4%, the consumption of L-HPC is 4%, the consumption of SLS is 1.5%.
Demonstration test
Mensuration is (52 ± 4) N according to prepared licoflavone dispersible tablet (n=3) hardness of above-mentioned preferred optimised process, and disintegration time is (72 ± 9) s, and can homodisperse.
(2) dispersing uniformity inspection
Dispersible tablet dispersing uniformity by the pharmacopeia regulation is checked, gets 2 of dispersible tablets that make and places 100mL water, is stirred to fully and disperses, and granule is all by No. 2 sieves.
(3) determination of dissolution rate
(a) preparation of reference substance solution
The accurate title, decided liquirtin reference substance 10mg, uses 50% dissolve with methanol, and standardize solution is in the 50mL volumetric flask, and getting concentration is 0.2mgmL
-1Reference substance solution.
(b) detect wavelength determination
Accurately draw liquirtin reference substance solution 2mL, add 7%NaOH solution 2mL, room temperature is placed 5min, to 10mL, does blank with methanol with methanol constant volume, and in the scanning of 190～800nm wavelength place, the result has absorption maximum at the 409nm place.With same method adjuvant is handled back scanning, find that adjuvant does not have absorption herein, thereby determine that detecting wavelength is 409nm.
(c) making of standard curve
Accurately draw 0.2mgmL
-1Liquirtin reference substance solution 0.05,0.1,0.25,0.5,1mL add 1mL methanol respectively, add 7%NaOH solution 2mL again, and room temperature is placed 5min, is diluted to 10mL with methanol, measure its absorption value at the 409nm place.With trap Y is vertical coordinate, and concentration X is an abscissa, gets equation of linear regression: Y=35.419X+0.0506, r=0.998, and the range of linearity is 1～20 μ gmL
-1
(d) assay method
According to the slurry method, measure the 300mL distilled water as dissolution medium, temperature is (37 ± 0.5) ℃, and rotating speed is (100 ± 1) rpm, respectively at 1,2,5,10,20, the learn from else's experience subsequent filtrate 2mL of 0.8 μ m filtering with microporous membrane of 30min, add 7%NaOH solution 2mL, room temperature is placed 5min, to 10mL, measures its absorption value with methanol constant volume at the 409nm place, and calculate its accumulation stripping percentage rate, the results are shown in Figure 1.
(e) licoflavone dispersible tablet stability study
The licoflavone dispersible tablet is at 40 ℃, and illumination is 4000Lx, and relative humidity is to place 3 months under 75% condition, and the content of licoflavone does not change (seeing Table 3 stability test results).
Table 3 licoflavone dispersible tablet stability test result (n=3)
Tab.7?The?results?of?stability(n＝3)
Experimental example 2 licoflavone dispersible tablets are done the application of medicament active composition in blood sugar lowering
(1) licoflavone dispersible tablet (LFT) is to the experimentation of mouse blood sugar influence
(1) the licoflavone dispersible tablet is to the blood sugar influence experiment of model induced by alloxan diabetic mice
Get 85 of Kunming mouses, male and female half and half, fasting 12h, except that normal control, each organizes equal lumbar injection alloxan (200ml/kg), the normal control group is a lumbar injection equivalent normal saline, predicts serum level of glucose behind the 72h, selects for use blood glucose value to test more than 16mmol/L, mice is divided into the normal control group, model group, acarbose group (25mg/kg), LFT high dose group (150ml/kg), dosage group (100mg/kg) among the LFT, LFT low dose group (50mg/kg), every day, gastric infusion was 1 time, and matched group and model group give the equal-volume distilled water, continuous 7 days, 1h after the last administration, the mouse orbit venous plexus is got blood, presses the determination of glucose oxidase blood sugar level.The results are shown in Table 4.
Table 4 LFT causes the influence of hyperglycemia mouse blood sugar to alloxan
Annotate: compare with model group
*P＜0.01, * p＜0.05
As can be seen from Table 4, compare with the alloxan model group, the large, medium and small dosage of LFT all raises to the alloxan diabetes mouse blood sugar remarkable inhibitory action.Compare effect with clinical use hypoglycemic medicine acarbose of the same type quite or better.
Overview: mice polydipsia, polyphagia, polyuria after the modeling, lose weight that promptly " three-many-one-little " symptom is obvious.Along with high, normal, basic three the dosage groups of LFT of carrying out of experiment are all alleviated to some extent to " three-many-one-little " symptom of model mice, especially amount of drinking water obviously reduces.
(2) the blood sugar influence experiment of licoflavone dispersible tablet hyperglycemia mice that epinephrine is caused
60 of Kunming mouses of experiment, male and female half and half, be divided into matched group at random, model group, acarbose group (25mg/kg), LFT high dose group (150ml/k), dosage group (100mg/kg) among the LFT, LFT low dose group (50mg/kg), matched group and model group give the equivalent normal saline, every day, gastric infusion was 1 time, continuous 7 days, 1h after the last administration, each group (except that the isopyknic normal saline of matched group lumbar injection) is lumbar injection epinephrine (240ug/kg) all, respectively at injection back 0.5 and 1h, from mouse orbit venous plexus measuring blood sugar of blood extracting content.The results are shown in Table 5.
Table 5 LFT causes the influence of hyperglycemia mouse blood sugar to epinephrine
Compare * P＜0.05, * * P＜0.01, * * *＜0.001 with the epinephrine group
The result shows, high, medium and low three the dosage groups of LFT have the adrenergic blood glucose increasing effect of obvious inhibition 0.5, during 1h.It is suitable with acarbose that it suppresses effect.
(3) the blood sugar influence experiment of licoflavone dispersible tablet hyperglycemia mice that glucose is caused
60 of Kunming mouses of experiment, male and female half and half, be divided into matched group at random, model group, acarbose group (25mg/kg), LFT high dose group (150ml/kg), dosage group (100mg/kg) among the LFT, LFT low dose group (50mg/kg), matched group and model group give the equal-volume normal saline, and every day, gastric infusion was 1 time, continuous 7 days, 1h after the last administration, remove matched group lumbar injection equal-volume normal saline, all the other respectively organize injectable dextrose monohydrate (2.0g/kg) solution, respectively behind injectable dextrose monohydrate 0.5,1,2h, from mouse orbit venous plexus measuring blood sugar of blood extracting content, the results are shown in Table 6.
Table 6 licoflavone dispersible tablet (LFT) causes the blood sugar influence of hyperglycemia mice to glucose
Compare * P＜0.05, * * P＜0.01, * * *＜0.001 with glucose group
As can be seen from Table 6, LFT has obvious inhibitory action to the blood sugar increasing that causes at the 0.5h glucose, and 1, still have certain blood sugar reducing function during 2h.Its hypoglycemic effect and acarbose are close.
Above-mentioned experimentation shows that the licoflavone dispersible tablet has significant hypoglycemic activity to multiple diabetic mice animal model, and middle dosage (100mg/kg) best results.
(2) safety research
(1) acute toxicity test
Trial test
Body weight 18-22g Kunming kind white mice, male and female half and half, totally 10 are used in test.Licoflavone dispersible tablet (LFT) is dissolved in a certain amount of distilled water, be made into Cmax (0.4g/ml) (not gambling pin with the filling stomach is as the criterion), once irritate stomach for mice and tried thing 0.8ml/20g, observe whether cause dead mouse, if can cause dead mouse, carry out LD by the preliminary experiment result
50Measure.If can not cause dead mouse, be limited to the administration volume, fail to measure its LD
50, carry out the maximum dosage-feeding experiment by pertinent regulations.
Result: behind the mouse stomach Cmax, do not cause that mice has unusual and the phenomena of mortality.
(2) maximum tolerated dose (MTD) experiment
Kunming kind white mice, 18-22g, male and female half and half, totally 20.Fasting is 16 hours before the experiment, can't help water, is tried thing 3 times by the maximum long-pending 0.8ml/20g of body of stomach that irritates of Cmax to mouse stomach in 24 hours, observes situations such as record mice crawler behavior, defecation and diet continuously seven days.Observed seven days, and do not see that mice had situations such as dystropy, death, body weight normal growth.Calculating its maximum filling stomach amount is 12g/kg.
Claims (2)
1, the preparation of licoflavone dispersible tablet, its step is the extracting liquorice decoction pieces, in 1: 4 ratio reflux, extract, of Radix Glycyrrhizae and 95% ethanol 3 times, each 2h, merging filtrate, reclaim solvent, get extractum, gained extractum is passed through 30-60 order polyamide column chromatography, water, 95% ethanol elution successively, with hydrochloric acid-magnesium powder reaction detection merge total flavones, again through concentrating and lyophilization gets the total flavones powder; Get 4-12% carboxymethyl starch sodium, 2-6%L-low substituted hydroxy-propyl fiber, 0.5-1.5% sodium lauryl sulphate and 20-85% licoflavone powder, add an amount of microcrystalline Cellulose, cross 100 mesh sieve mix homogeneously, after 60 ℃ of dryings, direct compression.
2, the licoflavone dispersible tablet is done the application of medicament active composition in blood sugar lowering.
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| CN102151296A (en) * | 2010-02-11 | 2011-08-17 | 兰州大学 | Method for extracting active ingredients for reducing sugar blood from waste residues of liquorice |
| CN104288218A (en) * | 2014-10-30 | 2015-01-21 | 王一杰 | Extracting method for glycyrrhiza flavonoid and application of glycyrrhiza flavonoid in preparation of medicine for treating liver-kidney injury caused by diabetes |
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| CN1203068C (en) * | 2003-07-11 | 2005-05-25 | 上海奥利实业有限公司 | Extraction and purification method of licoflavone |
| CN1879641A (en) * | 2005-06-18 | 2006-12-20 | 安徽省现代中药研究中心 | Dispersible tablet of glycyrrhizin and preparation method thereof |
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| CN102151296A (en) * | 2010-02-11 | 2011-08-17 | 兰州大学 | Method for extracting active ingredients for reducing sugar blood from waste residues of liquorice |
| CN102151296B (en) * | 2010-02-11 | 2012-05-09 | 兰州大学 | Method for extracting active ingredients for reducing sugar blood from waste residues of liquorice |
| CN104288218A (en) * | 2014-10-30 | 2015-01-21 | 王一杰 | Extracting method for glycyrrhiza flavonoid and application of glycyrrhiza flavonoid in preparation of medicine for treating liver-kidney injury caused by diabetes |
| CN105748428A (en) * | 2015-12-30 | 2016-07-13 | 惠州市九惠制药股份有限公司 | Preparation method of Anweiyang dispersible tablets |
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| CN106265572A (en) * | 2016-09-20 | 2017-01-04 | 南方医科大学 | A kind of 3D prints licoflavone dispersible tablet and preparation method thereof |
| CN106265572B (en) * | 2016-09-20 | 2019-03-05 | 南方医科大学 | A kind of 3D printing licoflavone dispersible tablet and preparation method thereof |
| CN106474083A (en) * | 2016-12-09 | 2017-03-08 | 宁夏回族自治区药品检验所 | Peaceful no sugar chawing tablet and its preparation technology and the application of Fructus Lycii sugar |
| CN106474083B (en) * | 2016-12-09 | 2019-05-17 | 宁夏回族自治区药品检验所 | Sweet Qi sugar is rather without sugar chawing tablet and its preparation process and application |
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Effective date of registration: 20190816 Address after: 222047 Jiangning economic and Technological Development Zone, Jiangsu, Lianyungang industrial city Patentee after: Kangyuan Pharmceutical Co., Ltd. Address before: 730000 Tianshui South Road, Gansu, Lanzhou, No. 222 Patentee before: Lanzhou University |