CN1175818C - Extractive preparation containing total alkali of mulberry leaves and its preparing method - Google Patents
Extractive preparation containing total alkali of mulberry leaves and its preparing method Download PDFInfo
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- CN1175818C CN1175818C CNB031019889A CN03101988A CN1175818C CN 1175818 C CN1175818 C CN 1175818C CN B031019889 A CNB031019889 A CN B031019889A CN 03101988 A CN03101988 A CN 03101988A CN 1175818 C CN1175818 C CN 1175818C
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Abstract
The present invention relates to a preparation containing mulberry leaf total base extractum and a preparing thereof, wherein the content of mulberry leaf total base in the extractum is not less than 70%, and the content of deoxynojirimycin is not less than 50%. The preparation can obviously lower blood sugar and improve sugar tolerance.
Description
Technical field
The present invention relates to a kind of Folium Mori total alkali extractum that contains, contain its preparation and preparation method thereof, said preparation has obvious functions of blood sugar and improves the carbohydrate tolerance effect.
Background technology
Folium Mori are the dried leaves of moraceae plants Mulberry Morous alba L..Folium Mori are the conventional Chinese medicine material, have dispelling wind and heat pathogens, clearing away lung-heat and moistening for dryness, the effect of liver heat removing and eyesight improving.The medical science man has made number of research projects to Folium Mori both at home and abroad, finds that the chemical constituent in the Folium Mori mainly contains flavonoid, sterols, polyhydroxylated alkaloid class, polysaccharide composition.The crude extract of Folium Mori, Folium Mori sterol (ketone), polysaccharides of Folium Mori and polyhydroxylated alkaloid show hypoglycemic activity respectively, and about the research of Folium Mori polyhydroxy biology total alkali, some reports are arranged both at home and abroad.N.Asano etc. have studied the chemical constituent of total alkali and the influence of monomer fagomine insulin secretion, and the Folium Mori total alkali is as an effective site, and main component content does not appear in the newspapers in the research of its hypoglycemic activity and the total alkali.The objective of the invention is to develop the polyhydroxylated alkaloid in the Folium Mori, develop a kind of high-load Folium Mori total alkali extractum.
Summary of the invention
The invention discloses a kind of high-load Folium Mori total alkali extractum, (English name is: 1-Deoxynojirimycin Hydrochloride) to contain a kind of alkaloid that is called deoxynojirimycin in the Folium Mori total alkali, it is a kind of main active, the Folium Mori total alkali that contains high level in the high-load Folium Mori total alkali extractum of the present invention, content is no less than 70% of extractum gross weight, and the content of the deoxynojirimycin in this Folium Mori total alkali is also higher.Its content is no less than 50% (percentage by weight) of extractum gross weight, and preparation method is fit to suitability for industrialized production.
High-load Folium Mori total alkali extractum of the present invention has obvious functions of blood sugar and improves the carbohydrate tolerance effect through the pharmacological research proof.
Another object of the present invention provides high-load Folium Mori total alkali extract making method of the present invention.
The preparation method of high-load Folium Mori total alkali is not seen bibliographical information, some steps are exhausted in research total alkali chemical constituent luck in the high-load Folium Mori total alkali extract making method of the present invention, the present invention also adopts some important step to improve the content of total alkali in the extractum except using these steps, concrete step is that Folium Mori coarse powder water or moisture hydrophilic solvent are extracted, make high-load Folium Mori total alkali extractum after concentrated, flocculation, column chromatography separation and purification, drying, wherein flocculation step has important function.
High-load Folium Mori total alkali extractum of the present invention prepares by the following method:
Folium Mori (50kg) powder is become coarse powder, cross 20 mesh sieves, water or moisture hydrophilic solvent heating and refluxing extraction with 500L, or the extraction of merceration percolation, heat is carried 2-3 time, each 1-3 hour, or cold extraction 1 time, 48-96 hour, the extracting solution concentrating under reduced pressure was proportion 1.12-1.20 then, added 1-5 times of water gaging dissolving of primary dose, leave standstill, regulate pH3～5, add flocculating agent chitosan 4L, leave standstill 12h, supernatant is the column chromatography of carrier by strong acid ion exchange resin, water is eluted to colourless, pours out resin, after adding that aqueous slkali is vexed and putting 4h, resin is extracted 2 times with the hydrophilic solvent methanol eddy, each 2h collects extracting solution, in being lower than concentrating under reduced pressure under 60 ℃ the condition, get high-load Folium Mori total alkali extractum 182g after the drying.
Described preparation method can be described with following steps:
A. dried mulberry leaf powder is broken into coarse powder, water or moisture hydrophilic solvent extract,
B. extracting solution concentrates, and is dissolved in water, and leaves standstill, and regulates pH to 3～5, adds flocculating agent, leave standstill,
C. supernatant is the column chromatography of carrier by ion exchange resin, and water is eluted to colourless, pours out resin, after adding that aqueous slkali is vexed and putting 4h, resin with hydrophilic solvent reflux, extract, (2h * 2), is collected extracting solution,
D. concentrating under reduced pressure, extractum.
Hydrophilic solvent described in the high-load Folium Mori total alkali extract making method of the present invention is aliphatic lower alcohol or lower ketones.
Wherein said flocculating agent is gelatin, sodium alginate, agar, chitosan and derivant thereof, ZTC1+1 etc., preferred chitosan.
Alkali in the wherein said aqueous alkali is volatile alkali, can be preferably ammonia for diethylamino, triithylamine, pyridine etc., and the mixed proportion of water and alkali is: water: ammonia=99-1: the arbitrary proportion of 1-99 (volume ratio).
High-load Folium Mori total alkali extract making method of the present invention is easy and simple to handle, does not have harsh equipment requirements, is suitable for the scale suitability for industrialized production.
High-load Folium Mori total alkali extractum of the present invention, can add the medicine acceptable carrier when being prepared into medicament, described medicine acceptable carrier can be: mannitol, silicon derivative, cellulose and derivant thereof, alginate, gelatin, polyvinylpyrrolidone, glycerol, soil temperature 80, agar, calcium carbonate, calcium bicarbonate, surfactant, Polyethylene Glycol, cyclodextrin, beta-schardinger dextrin-, phospholipid material, Kaolin, Pulvis Talci, calcium stearate, magnesium stearate etc.
High-load Folium Mori total alkali extractum of the present invention can be made Chinese medicine preparation by conventional method, said preparation can be any pharmaceutically useful dosage form, and these dosage forms comprise: tablet, sugar coated tablet, film coated tablet, enteric coated tablet, capsule, hard capsule, soft capsule, oral liquid, suck agent, granule, electuary, pill, powder, unguentum, sublimed preparation, suspensoid, powder, solution, injection, suppository, ointment, plaster, cream, spray, drop, patch.Preparation of the present invention, peroral dosage form preferably, as: capsule, tablet, oral liquid, granule, pill, powder, sublimed preparation, unguentum etc., preferably capsule.
Can contain Folium Mori total alkali extractum 10-600mg in every dose in the preparation of the present invention, described every dose of finger, each preparation unit, or each taking dose, if each preparation unit can refer to every capsules, or every, every bag, every ball, each peace bottle etc.
Determine usage and dosage according to patient's situation in use, but as obeying every day three times, each 1-20 agent, as: 1-20 grain or sheet.
Extractum of the present invention is carried out following pharmacodynamic experiment proved its beneficial effect.
One, to the influence of normal mouse blood sugar
Test material
1, medicine: the Folium Mori total alkali with the preparation of embodiment 1 method, is mixed with needed concentration with distilled water before the test.
2, animal: NIH mice, 18-22g, ♀, ♂ half and half.
3, blood sugar detection test kit is produced lot number 20020306 by Sichuan Michelson's thing technology company limited.
Test method and result
1, single-dose is to the influence of blood glucose: mice, 30, on an empty stomach behind the 12h, by animality not equilibrium be divided into 3 groups at random, 10 every group, ♀, ♂ half and half irritate stomach respectively and give following medicine: (1) normal control group (distilled water); (2) Folium Mori total alkali low dose group (40mg/kg); (3) Folium Mori total alkali high dose group (100mg/kg).The administration volume is pressed 20ml/kg, and animal eyeball is got blood behind the administration 1h, uses the determination of glucose oxidase fasting blood sugar.The results are shown in Table 1.
Table 1 Folium Mori total alkali (single-dose) is to the influence of normal mouse blood sugar value
(n＝10，X±SD)
The dosage blood glucose value
Group
(mg/kg) (mmol/L)
Normal control group-1.797 ± 0.566
Folium Mori total alkali 40 0.655 ± 0.536**
Folium Mori total alkali 100 0.929 ± 0.904*
Compare * P＜0.05, * * P＜0.01 (t check) with the normal control group
2, multiple dosing is to the influence of blood glucose: mice, and 20, ♀, ♂ half and half behind the 12h, measure blood glucose on an empty stomach, are divided into 2 groups at random according to blood glucose value and the equilibrium of animal sex, irritate stomach respectively and give following medicine: (1) normal control group (distilled water); (2) Folium Mori total alkali (80mg/kg).Successive administration 7 days, the administration volume is pressed 20ml/kg, empty stomach 12h before the administration in the 7th day, the 1h animal is got blood after the administration, measures the animal blood glucose value with happy one-tenth blood glucose monitoring system.The results are shown in Table 2.
Table 2 Folium Mori total alkali (multiple dosing) is to the influence of normal mouse blood sugar value
(n＝10，X±SD)
Blood glucose value rate of change behind the blood glucose value medicine before the dosage medicine
Group
(mg/kg) (mmol/L) (mmol/L) (％)
Normal control group-4.74 ± 0.89 4.95 ± 0.85+6.9 ± 22.4
Folium Mori total alkali 80 4.67 ± 1.05 3.56 ± 1.17**-22.9 ± 21.4
Compare * P＜0.05, * * P＜0.01 (t check) with the normal control group.
Conclusion
1, single-dose, and the Folium Mori total alkali (40,100mg/kg), have the effect of tangible blood sugar lowering.
2, multiple dosing, Folium Mori total alkali (80mg/kg) has the effect of blood sugar lowering.
Two, to the influence of spontaneous hyperglycemia mouse blood sugar
Test material
1, medicine: the Folium Mori total alkali, with the preparation of embodiment 1 method.Be mixed with needed concentration with distilled water before the test.
2, animal: the NOD mice, body weight 20-30g, 24, ♀, ♂ half and half are provided by Department Of Medicine, Peking University's Experimental Animal Center, licence numbering: SCXK11-00-0004.
3, determining instrument: happy one-tenth blood glucose monitoring system and mensuration reagent paper, develop by Yicheng Biological Electronic Technology Co., Ltd., Beijing.
Test method and result
Mice, 24, on an empty stomach behind the 12h, measure blood glucose value,, the animal equilibrium is divided into 3 groups at random according to blood sugar level and sex, every group 8, ♀, ♂ half and half irritate stomach respectively and give following medicine: (1) matched group (distilled water), (2) Folium Mori total alkali low dose group (50mg/kg), (3) Folium Mori total alkali high dose group (150mg/kg), the administration volume is pressed 20ml/kg, behind the administration 1h, animal is got blood, measures fasting blood sugar with happy one-tenth blood glucose monitoring system.The results are shown in Table 3.
Table 3 Folium Mori total alkali is to the influence of spontaneous hyperglycemia mouse blood sugar
(n＝8，X±SD)
Blood glucose rate of change behind the blood glucose medicine before the dosage medicine
Group
(mg/kg) (mmol/L) (mmol/L) (％)
Matched group-8.61 ± 1.51 8.99 ± 1.42+5.0 ± 9.5
Folium Mori total alkali 50 8.48 ± 1.38 7.83 ± 1.41-7.1 ± 12.5
Folium Mori total alkali 150 8.79 ± 1.76 8.20 ± 1.20-5.6 ± 8.3
Medicine group and matched group compare, and difference does not have significance meaning P＞0.05 (t check).
Conclusion
The Folium Mori total alkali does not have obvious influence to spontaneous hyperglycemia mouse blood sugar.
Three, streptozotocin is brought out the influence of mice hyperglycemia
Test material
1, medicine: the Folium Mori total alkali with the preparation of embodiment 1 method, is mixed with needed concentration with distilled water before the test.
2, animal: NIH mice, body weight 18-22g, ♀, ♂ half and half.
3, main agents: streptozotocin, U.S. Sigma company product.With citrate buffer solution (PH=4.0), prepare down before the modeling in low temperature.
4, instrument: happy one-tenth blood glucose monitoring system and mensuration reagent paper, develop by Yicheng Biological Electronic Technology Co., Ltd., Beijing.
Test method and result
Mice, 40, divide 42 groups at random, 10 every group, ♀, ♂ half and half, (1) matched group (distilled water); (2) hyperglycemia model group (distilled water); (3) Folium Mori total alkali low dose group (50mg/kg); (4) Folium Mori total alkali high dose group (150mg/kg).Except that the normal control group, all the other respectively organize lumbar injection streptozotocin (200mg/kg), survey blood glucose (12h on an empty stomach) behind the 72h, then with (2)-(4) group, according to the balanced group of adjusting of blood glucose value.Irritate stomach by above dosage simultaneously and give trial drug, continuous 7 days, empty stomach 12h before the administration in the 7th day, 1h measures blood glucose after the last administration.The results are shown in Table 4.
Table 4 Folium Mori total alkali brings out the influence of mice hyperglycemia to streptozotocin
(n＝10，X±SD)
Blood glucose rate of change behind the blood glucose medicine before the dosage medicine
Group
(mg/kg) (mmol/L) (mmol/L) (％)
Normal control group-4.73 ± 1.01 4.86 ± 1.02+3.1 ± 7.8
Hyperglycemia model group-9.34 ± 2.23
##9.49 ± 2.04
##+ 2.1 ± 5.8
Folium Mori total alkali 50 9.60 ± 2.68 7.59 ± 2.41-21.3 ± 5.5
Folium Mori total alkali 150 9.76 ± 2.60 7.39 ± 1.80*-23.8 ± 7.8
Compare with the normal control group
##P＜0.01; Compare * P＜0.05, * * P＜0.01 (t check) with model group.
Conclusion
Mouse peritoneal injection streptozotocin can make blood sugar increasing, and after giving the Folium Mori total alkali, have hypoglycemic effect.Wherein dosage is that (150mg/kg) compares with model group, and difference has the significance meaning.
Four, to the carbohydrate tolerance test of the oral sucrose of mice
Test material
1, medicine: the Folium Mori total alkali with the preparation of embodiment 1 method, is mixed with needed concentration with distilled water before the test.
2, animal: NIH mice, 18-22g, ♀, ♂ half and half.
3, main agents: sucrose, analytical pure, Guangzhou Chemical Reagent Factory production, lot number 960202.
4, instrument: happy one-tenth blood glucose monitoring system and mensuration reagent paper, develop by Yicheng Biological Electronic Technology Co., Ltd., Beijing.
Test method and result
Mice, 20, be divided into 2 groups at random, 10 every group, ♀, ♂ half and half irritate stomach respectively and give following medicine: (1) matched group (distilled water); (2) Folium Mori total alkali group (80mg/kg), the administration volume is 20ml/kg, behind the animal empty stomach 12h, measures blood glucose, then by above dosed administration, and contains sucrose 0.25g/ml in the medicine.Measure 1h, 2h blood glucose value to 0.5h after the sucrose with happy one-tenth blood glucose monitoring system.Result of the test sees Table 5.
Table 5 Folium Mori total alkali is to the carbohydrate tolerance test result
(n＝10，X±SD)
Dosage blood glucose value (mmol/L) rate of change (%)
Group
(mg/kg) 0h 0.5h 1h 2h 0.5h 1h 2h
Normal control-4.66 ± 7.72 ± 8.10 ± 5.08 ± 69.0 ± 78.0 ± 11.3 ±
Organize 1.01 1.05 0.97 0.79 18.4 23.0 16.7
4.50± 6.73± 7.18± 4.68± 56.2± 69.2±
Folium Mori total alkali 80 5.6 ± 9.3
1.18 1.01* 0.75* 1.08 31.7 43.1
With relatively * P＜0.05 (the t check) of normal control group.
Conclusion
Folium Mori total alkali, dosage are the rising that 80mg/kg can suppress blood glucose value after the oral sucrose of mice.
The specific embodiment:
Further specify the present invention by the following examples
Embodiment 1,
The dry coarse powder 1kg of Folium Mori crosses 20 mesh sieves, adds 20% aqueous acetone solution 10kg, heating and refluxing extraction 2h, filter, repeat heat and carry totally three times, the extracting solution concentrating under reduced pressure is a proportion 1.20, add water 3kg dissolving, leave standstill, regulate pH3～5, add an amount of 1% chitosan with hydrochloric acid, leave standstill 12h, supernatant is the column chromatography of carrier by cation exchange resin 1kg, and water 3kg eluting is abandoned or adopted, pour out resin, after adding that aqueous slkali is vexed and putting 4h, with resin with 80% alcohol reflux twice, each two hours, collect extracting solution, in being lower than concentrating under reduced pressure under 60 ℃ the condition, get high-load Folium Mori total alkali extractum 4g (deoxynojirimycin content is 57%) after the drying.
Embodiment 2,
The dry coarse powder 10kg of Folium Mori, add 20% ethanol 80kg, heating and refluxing extraction 1.5h filters, repeating heat carries three times totally, the extracting solution concentrating under reduced pressure is a proportion 1.15, adds water 40kg dissolving, regulates pH3～5 with 10% hydrochloric acid, add an amount of 1% chitosan, leave standstill 12h, supernatant is the column chromatography of carrier by cation exchange resin 10kg, water 30kg eluting, abandon or adopt, supernatant is the column chromatography of carrier by ion exchange resin, and water is eluted to colourless, pours out resin, after adding that aqueous slkali is vexed and putting 4h, resin is extracted 2 times with the hydrophilic solvent methanol eddy, and each 2h collects extracting solution.In be lower than under 60 ℃ the condition after concentrating under reduced pressure, the drying high-load Folium Mori total alkali extractum 37g (deoxynojirimycin content is 55%).
Embodiment 3,
The dry coarse powder 100kg of Folium Mori adds water 1000kg, heating and refluxing extraction 2h, filter, repeat heat and carry totally three times, the extracting solution concentrating under reduced pressure is a proportion 1.2, add water 2500kg dissolving, regulate pH3～5, add an amount of 1% chitosan with 10% hydrochloric acid, leave standstill 12h, supernatant is the column chromatography of carrier by D001 type cation exchange resin 1000kg, water 3000kg eluting, pour out resin, after adding that aqueous slkali is vexed and putting 4h, resin is extracted 2 times with the hydrophilic solvent methanol eddy, each 2h collects extracting solution.In be lower than under 60 ℃ the condition after concentrating under reduced pressure, the drying high-load Folium Mori total alkali extractum 320g (deoxynojirimycin content is 52%).
Embodiment 4
Capsule
[prescription] Folium Mori total alkali extractum 60g
Make 1000 of capsules
[method for making] with Folium Mori total alkali extractum 60g, adds mixings such as pharmaceutic adjuvant microcrystalline Cellulose, granulates, and drying incapsulates and gets final product.
Embodiment 5
Tablet
[prescription] Folium Mori total alkali extractum 60g
Make 1000 in tablet
[method for making] adds mixings such as pharmaceutic adjuvant microcrystalline Cellulose with Folium Mori total alkali extractum 60g, granulates, and drying adds the moderate lubrication agent, and tabletting gets final product.
Embodiment 6
Granule
[prescription] Folium Mori total alkali extractum 100g
Make 1000 bags of granules
[method for making] adds mixings such as pharmaceutic adjuvant dextrin, sweeting agent with Folium Mori total alkali extractum 100g, granulate, and drying, pack gets final product.
Embodiment 7
Pill
[prescription] Folium Mori total alkali extractum 100g
Make 1000 bags of pills
[method for making] adds general balls such as pharmaceutic adjuvant dextrin, microcrystalline Cellulose with Folium Mori total alkali extractum 100g, drying, and packing gets final product.
Embodiment 8
Oral liquid
[prescription] Folium Mori total alkali extractum 100g
Make 1000 of oral liquids
[method for making] is dissolved in water with Folium Mori total alkali extractum 100g, adds an amount of correctives, sterilization, and fill gets final product.
Claims (7)
1, a kind of Folium Mori total alkali extractum contains the effective ingredient deoxynojirimycin, and the content of Folium Mori total alkali is no less than 70% in this extractum, and the content of deoxynojirimycin is no less than 50%, it is characterized in that, this extractum prepares through following steps,
A. dried mulberry leaf powder is broken into coarse powder, water or moisture hydrophilic solvent extract;
B. extracting solution concentrates, and is dissolved in water, and leaves standstill, and regulates pH to 3～5, adds flocculating agent, leaves standstill;
C. supernatant is the column chromatography of carrier by ion exchange resin, and water is eluted to colourless, pours out resin, after adding that aqueous slkali is vexed and putting 4h, resin with hydrophilic solvent reflux, extract, (2h * 2), is collected extracting solution;
D. concentrating under reduced pressure, extractum.
2, extractum according to claim 1 is characterized in that, described hydrophilic solvent is aliphatic lower alcohol or lower ketones.
3, extractum according to claim 1 is characterized in that, described flocculating agent is selected from gelatin, sodium alginate, agar, chitin and derivant thereof, chitosan.
4, extractum according to claim 1 is characterized in that, the alkali in the described aqueous slkali is volatile alkali, is selected from diethylamino, triithylamine, pyridine, ammonia, and the mixed proportion of water and alkali is: 99-1: 1-99.
5, use the pharmaceutical preparation of the extractum preparation of claim 1.
6, the pharmaceutical preparation of claim 5 is any pharmaceutically useful dosage forms.
7, the application of the extractum of claim 1 in a kind of medicine for the treatment of diabetes of preparation.
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| CNB031019889A CN1175818C (en) | 2003-01-30 | 2003-01-30 | Extractive preparation containing total alkali of mulberry leaves and its preparing method |
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| CN1175818C true CN1175818C (en) | 2004-11-17 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100464764C (en) * | 2006-03-08 | 2009-03-04 | 江苏大学 | Composition of white mulberry leaf blood-sugar-reducing effective components and preparing method |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1312117C (en) * | 2004-12-03 | 2007-04-25 | 上海丝绸集团股份有限公司 | Extraction technology of gamma amino butyric acid in folium mori |
| CN100457106C (en) * | 2005-06-15 | 2009-02-04 | 同济大学 | Use of 1-deoxynojirimycin for preparing diabete and disney disease drug |
| CN101671294B (en) * | 2009-09-22 | 2011-12-21 | 广东太阳神集团有限公司 | Method for continuously extracting and separating 1-deoxynojirimycin (DNJ) and flavone from folium mori |
| CN103623059B (en) * | 2013-12-04 | 2016-01-06 | 湖南希尔天然药业有限公司 | A kind of pharmaceutical composition and application thereof, containing its medicament |
| CN104557675A (en) * | 2014-12-08 | 2015-04-29 | 郑森衔 | Method for preparing 1-deoxynojirimycin |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100464764C (en) * | 2006-03-08 | 2009-03-04 | 江苏大学 | Composition of white mulberry leaf blood-sugar-reducing effective components and preparing method |
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