CN1605357A - Fenugreek seed extract and its preparing process and application - Google Patents
Fenugreek seed extract and its preparing process and application Download PDFInfo
- Publication number
- CN1605357A CN1605357A CN 200410070104 CN200410070104A CN1605357A CN 1605357 A CN1605357 A CN 1605357A CN 200410070104 CN200410070104 CN 200410070104 CN 200410070104 A CN200410070104 A CN 200410070104A CN 1605357 A CN1605357 A CN 1605357A
- Authority
- CN
- China
- Prior art keywords
- extract
- semen trigonellae
- polysaccharide
- saponin
- trigonellae extract
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
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Landscapes
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention discloses one kind of fenugreek extractive, its preparation process, medicine composition and application. The extractive contains three kinds of active components, including polysaccharide 15-50 wt%, total flavone 5-15 wt% and saponin 10-30 wt%, and has obvious blood sugar and blood fat reducing effect and thus high treating effect on diabetes and its complications.
Description
Technical field
The present invention relates to a kind of Semen Trigonellae extract and preparation method thereof, and the pharmaceutical composition that contains this Semen Trigonellae extract, described extract has the very strong blood sugar lowering and the activity of blood fat reducing, and this extract and pharmaceutical composition thereof are mainly used in treatment diabetes and hyperlipidaemic conditions.
Background technology
Semen Trigonellae is the dry mature seed of pulse family annual herb plant Semen Trigonellae Trigonella foenum-graecum, and this medical material originates in Arabic area, and the Tang Dynasty is imported China into.Warm in nature, bitter in the mouth is nontoxic; Has kidney-replenishing, the effect of dispelling cold and dampness; Normal single or compound recipe is used for the treatment of colic of cold type, the disturbance of lower legs due to pathogenic cold and dampness, the soreness of waist of suffering from a deficiency of the kidney and sexual impotence etc. in the traditional Chinese medical science; Recorded by 2000 editions pharmacopeia.As external medicinal plants, Eastern Mediterranean riparian land district during Semen Trigonellae originates in is in the Middle East, areas such as Africa and India are as food and the existing long history of medicine.The equal edible of seedling of Semen Trigonellae and seed, seed are that the Indian is extensively edible as a kind of curried flavoring agent; As the folk tradition medicine, Semen Trigonellae Chang Zuowei nourishing, strong medicine, aperitive and antipyretic analgesics use.
Different with traditional use habit, studies show that at present fenugreek seed has significant blood sugar lowering and effect for reducing blood fat, the modern study direction also is primarily aimed at the blood sugar lowering of fenugreek seed and the pharmacological action of blood fat reducing aspect.For example, bibliographical information is abroad arranged, the Semen Trigonellae skimmed milk has significantly reduced experimenter's cholesterol, triglyceride and very low density lipoprotein (VLDL) value, its result of the test shows that Semen Trigonellae can effectively regulate the blood lipid level that improves hyperlipemic patients, but more than being the former powder of medicine is used as medicine, dosage is subjected to the crude drug quality control, and effective ingredient is smudgy, is difficult to quantitatively.
Application number is that 03144071.1 patent application has disclosed a kind of Semen Trigonellae extract, and it is to be the correlational study that the unique active component of extract (being effective ingredient) carries out blood sugar lowering and blood fat reducing with the saponin.In fact, have the Semen Trigonellae extract that only limits to contain single saponin absolutely not of blood sugar lowering and effect for reducing blood fat, but contain the effective site of polysaccharide, total flavones and saponin at least.Experiment showed, that its drug effect is had a greatly reduced quality if do not contain or only contain a spot of polysaccharide, total flavonoid in the Semen Trigonellae extract.
At present domestic and international present Research about Semen Trigonellae, has abundant Semen Trigonellae herb resource with respect to China, crude drug is cheap, the huge present situation of while diabetics crowd, the external report that the use skimmed milk is only arranged, therefore further study Semen Trigonellae extract both in depth and in breadth, should have a good application prospect.
Summary of the invention
Through a large amount of research, the inventor has obtained a kind of Semen Trigonellae extract with notable biological activity, and it is little that Semen Trigonellae extract has dosage, and effect is significantly carried, characteristics such as taking convenience.
The purpose of this invention is to provide a kind of Semen Trigonellae extract, it contains polysaccharide, the total flavones of 5-15% and the saponin of 10-30% that percentage by weight is respectively 15-50%, and this extract has the effect of blood sugar lowering and blood fat reducing.
The present invention also aims to provide a kind of method for preparing Semen Trigonellae extract, extract by adopting alcohol/aqueous solvent, concentrating and impurity removing, dry and extract have remarkable blood sugar lowering, effect for reducing fat.
The present invention also aims to provide a kind of pharmaceutical composition that contains above-mentioned Semen Trigonellae extract, it has the effect of blood sugar lowering, blood fat reducing.
The present invention also aims to provide the application of Semen Trigonellae extract in the medicine of preparation treatment diabetes and hyperlipidemia.
The present invention also aims to provide a kind of application of pharmaceutical composition in treatment diabetes and hyperlipidemia that contains Semen Trigonellae extract.
The invention provides a kind of Semen Trigonellae extract, it contains polysaccharide, the total flavones of 5-15% and the saponin of 10-30% that percentage by weight is respectively 15-50%.
The present invention also provides the preparation method of above-mentioned Semen Trigonellae extract, after the Semen Trigonellae pulverizing medicinal materials, is the pure water mixed solvent of 20%-90% with the determining alcohol, extracts.
The present invention also provides the application of Semen Trigonellae extract in the medicine of preparation treatment diabetes and hyperlipidemia.
The present invention also provides a kind of pharmaceutical composition, and it contains the above-mentioned Semen Trigonellae extract and the pharmacy acceptable auxiliary for the treatment of effective dose.
The present invention also provides the application of aforementioned pharmaceutical compositions in treatment diabetes and hyperlipidemia.
Semen Trigonellae extract of the present invention contains polysaccharide, flavone and saponin, these three kinds of main active substances have remarkable blood sugar lowering, effect for reducing fat because of synergism together, extract except the pure water percolation of routine in the preparation process of this extract and remove impurity, also carried out effective enrichment, made above-mentioned three kinds of effective ingredient be present in the extract with certain content.
Semen Trigonellae extract of the present invention, wherein each weight percentages of components is preferably: polysaccharide 40%, total flavones 10%, saponin 20%; Wherein said content of total flavone is 5-15%, and sign property composition is an orientin, and the content of orientin is 0.3-1.2% in this extract, preferred 0.7%; Total saponins is 10-30% in the content of diosgenin.This extract has the effect of blood sugar lowering and blood fat reducing.
The preparation method of Semen Trigonellae extract of the present invention is: after the Semen Trigonellae pulverizing medicinal materials, extract with pure water mixed solvent, this extracting method can be any conventional extracting method, preferred percolation, wherein alcohol can be lower alcohols such as methanol, ethanol, preferred alcohol, and concentration of ethanol is 20%-95%, preferred 50-75% makes with extra care and gets through enrichment again.
Pharmaceutical composition of the present invention, it contains the Semen Trigonellae extract and the pharmacy acceptable auxiliary for the treatment of effective dose, and this Semen Trigonellae extract contains polysaccharide, the total flavones of 5-15% and the saponin of 10-30% that percentage by weight is respectively 15-50%; Preferred each weight percentages of components is preferably: polysaccharide 40%, total flavones 10%, saponin 20%.
Above-mentioned composition of the present invention, it can be any peroral dosage form or injection type, preferred oral dosage form, more preferably tablet, capsule or granule.
Above-mentioned preferred dosage form, tablet for example, it contains the above-mentioned Semen Trigonellae extract and the pharmacy acceptable tablet adjuvant for the treatment of effective dose, wherein the content of this Semen Trigonellae extract in compositions is 0.1～99.9% (weight), be preferably 40～90% (weight), 65～75% (weight) more preferably, with described adjuvant be conventional adjuvant, for example be selected from microcrystalline Cellulose, low-substituted hydroxypropyl cellulose, cross-linking sodium carboxymethyl cellulose, pregelatinized Starch, micropowder silica gel and the magnesium stearate at least a.
Aforementioned pharmaceutical compositions can be used for treating diseases such as diabetes and hyperlipidemia.
The inventor adopts alcohol/aqueous solvent to extract in extraction process at the three class main active substances polysaccharide, flavone and the saponin that contain in the Semen Trigonellae, and concentrating and impurity removing has carried out effective enrichment with this three classes material simultaneously, and concrete enrichment method is as follows:
With the dry medicinal material coarse powder 50-75% ethanol percolate extraction of Semen Trigonellae, percolate concentrate under reduced pressure at low temperature to relative density is about 1.15-1.20, can adopt 3 kinds of distinct methods to make with extra care respectively:
(1) extracting solution extracts with the equivalent water-saturated n-butanol, and extract reclaims the solvent after drying and gets extract.
(2) extracting solution ethyl acetate extraction defat, the mother solution convection drying gets extract.
(3) extracting solution carries out purification with the D101 resin column, washes with water to remove the high polarity polysaccharose substance, uses 95% ethanol elution then, collects eluent, and the decompression and solvent recovery after drying promptly gets extract.
Can obtain solids extraction thing of the present invention by above-mentioned process for purification, drying mode can be any conventional drying method, preferably spray drying.
Through a large amount of experiments, above-mentioned three kinds of Semen Trigonellae extracts have been obtained, wherein polysaccharide content is few in the extract that obtains of method (1) and (3), (2) extract that method obtains then contains more polysaccharide, total flavones and saponin, in conjunction with pharmacological testing, the result proves that the method for (2) acquires extract than other two kinds of methods more significant blood sugar lowering is arranged, the blood fat reducing activity, the polysaccharide compound that high polarity in the Semen Trigonellae is described also is its indispensable effective ingredient, experimental results show that: polysaccharide, total flavones and saponin synergism are only Semen Trigonellae extract and bring into play its blood sugar lowering, the active key point of blood fat reducing.
Preferably, enrichment method employing method of the present invention (2).
In fact, various theories are also instructed, polysaccharide has significant blood sugar reducing function, saponins then has significant effect for reducing fat, flavonoid can microcirculation improvement, the multiple complication of diabetics is had regulating action, therefore, the therapeutical effect of Semen Trigonellae is the result of the collaborative many target spot regulation and control of this three constituents, and is indispensable.
By the systematic study to this extract quality, the feature of Semen Trigonellae extract is as described below:
1, polysaccharose substance
The assay of polysaccharose substance adopts the ultraviolet derivatization method, and concrete operations are as follows:
The preparation of reference substance solution: get the about 36mg of glucose of 80 ℃ of dry constant weights, the accurate title, decide, and puts in the 100ml measuring bottle, and thin up is to scale; Shake up, precision is measured 10ml, puts in 100 measuring bottles, and thin up shakes up to scale, promptly gets (every 1ml contains glucose 36 μ g).
The preparation of need testing solution: get Semen Trigonellae extract, porphyrize, precision claims to conclude a contract or treaty puts 450mg in the 50ml measuring bottle, adds suitable quantity of water, and ultrasonic 5min is chilled to room temperature, and thin up shakes up to scale; Precision is measured subsequent filtrate 2ml, crosses macroporous resin DIAION HP20 post (diameter 1.0cm, height 20cm), the water eluting, and 100ml eluent before collecting shakes up, promptly.
Algoscopy: precision is measured reference substance solution, sample solution 2ml respectively, puts in the 10ml tool plug test tube, adds 5% phenol solution of 1.0ml, mixing; Add the 5.0ml concentrated sulphuric acid then, mixing, static, survey its trap (operate with water simultaneously in accordance with the law and do blank) at the 490nm place, calculate content.
Conclusion: the content of polysaccharide is 15-50% in the middle of the Semen Trigonellae extract.
2, saponins material
The mensuration of saponins material adopts the ultraviolet derivatization method, the saponins content assaying method is from relevant references (Pierre R.Petit etc., Yves D. Sauvaire, et al.Steroidsaponinsfrom fenugreek seeds:Extraction, purification, and pharmacologicalinvestigation on feeding behavior and plasma cholesterol.Steroids.1995,60:674-680), specific as follows:
The preparation of reference substance solution: precision takes by weighing diosgenin reference substance 20mg, puts in the 100ml measuring bottle, adds ethanol and dissolves in right amount and be diluted to scale, shake up, precision is measured 10ml, puts in the 100ml measuring bottle, add ethanol dilution to scale, shake up, promptly get (every 1ml contains diosgenin 0.02mg).
The preparation of need testing solution: get Semen Trigonellae extract, porphyrize, precision takes by weighing about 20mg, puts in the 100ml measuring bottle, adds water 10ml, and ultrasonic 5 minutes, add ethanol dilution to scale after the placement room temperature, shake up promptly.
The preparation of test solution: 0.5% anisaldehyde ethyl acetate solution; 50% sulphuric acid ethyl acetate solution.
Algoscopy: precision is measured test sample and reference substance solution 2ml respectively, puts in the 10ml tool plug test tube evaporate to dryness, after treating that test tube is chilled to room temperature, the accurate Glacial acetic acid ethyl ester 2ml that adds adds 1ml 0.5% anisaldehyde ethyl acetate solution then, and 1ml 50% sulphuric acid ethyl acetate solution; Behind the solution mixing, put in 60 ℃ of water-baths and reacted 30 minutes, take out and be cooled to room temperature, add 5ml anhydrous acid ethyl acetate, shake up.Survey the trap at 430nm place, calculate content.
Conclusion: the content of saponin is 10-30% in the middle of the Semen Trigonellae extract.
3, total flavonoids substance
The determination of total flavonoid composition is as follows:
The preparation of reference substance solution: the accurate title, decided the about 10mg of orientin reference substance, puts in the 100ml measuring bottle, adds methanol to scale, shakes up, and precision is measured 10ml, puts in the 100ml measuring bottle, adds methanol to scale, shakes up, and promptly gets (every 1ml contains orientin 0.01mg).
The preparation of need testing solution: get this product, porphyrize, precision takes by weighing about 100mg, puts in the 100ml measuring bottle, and adding methanol is an amount of earlier, ultrasonic 5min adds methanol then to scale, shakes up, and precision is measured subsequent filtrate 10ml, put in the 100ml measuring bottle, add methanol, shake up, promptly to scale.
Algoscopy: measure reference substance solution and need testing solution trap, calculate at the 334nm place.
Conclusion: content of total flavone is 5-15% in the Semen Trigonellae extract.
4, the discrimination test of the dioscin constituents in the middle of the Semen Trigonellae extract
The saponins material that contains in the middle of the Semen Trigonellae is mainly the derivant of dioscin, and its aglycon is mainly diosgenin, by the description Semen Trigonellae extract of differentiating that dioscin constituents in the middle of the extract can be comparatively clear and definite.The reference substance diosgenin is available from Chinese biological goods calibrating institute.
Discrimination test is as described below:
An amount of porphyrize of Semen Trigonellae extract is got the powder of about 0.5g, puts in the 150ml round-bottomed flask, adds 6% ethanol solution of sulfuric acid 50ml and (gets sulphuric acid 60ml, slowly inject 40% an amount of alcoholic solution, put coldly, add 40% ethanol to 1000ml, shake up), ultrasonic 5 minutes, filter, filtrate is put in the boiling water bath and to be refluxed 5 hours, puts coldly, adds water 100ml, shake up, filter, residue is washed with water to filtrate and does not show acid, 105 ℃ of oven dry, with the dissolving of 5ml ethyl acetate, solution is as need testing solution.Other gets the diosgenin reference substance, adds ethyl acetate and makes the solution that every 1ml contains diosgenin reference substance 2mg, in contrast product solution.According to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia D) test, draw each 5 μ l of above-mentioned two kinds of solution and put respectively on same silica gel g thin-layer plate, with petroleum ether: ethyl acetate (5: 1) is developing solvent, launch, take out, dry, spray 3% phosphomolybdic acid ethanol solution, 105 ℃ were heated several minutes.In the test sample chromatograph,, show the speckle of same color in the corresponding position of reference substance chromatograph.
5, the discriminating of orientin in the middle of the Semen Trigonellae extract
Flavonoid substances in the middle of the Semen Trigonellae is mainly flavonoid glycoside, and orientin is differentiated the orientin in the middle of the Semen Trigonellae extract by the HPLC method for its sign property composition.
Measure with high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia D).With octadecylsilane chemically bonded silica is filler; Acetonitrile-isopropyl alcohol-water (8: 4: 88) (regulating pH value to 3.2 with phosphoric acid) is a mobile phase; The detection wavelength is 334nm.Number of theoretical plate calculates by orientin should be not less than 3000.
The preparation of reference substance solution: precision takes by weighing orientin reference substance (self-control) 10mg, puts in the 100ml measuring bottle, adds methanol to scale, shakes up; Precision is measured 5ml, puts in the 10ml measuring bottle, adds methanol to scale, shakes up, and promptly gets (every 1ml contains orientin 0.05mg).The preparation of need testing solution: get this product under the weight differential item, porphyrize, mixing, precision takes by weighing about 500mg, and to the 100ml measuring bottle, it is an amount of to add entry, and ultrasonic 5 minutes, add methanol to scale after the placement room temperature, shake up, promptly.
Accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing inject chromatograph of liquid promptly.Record orientin assay chromatogram, orientin is consistent with the retention time of reference substance in the sample.
The content of orientin is 0.3-1.2% in the Semen Trigonellae extract of the present invention.
Pharmaceutical composition provided by the invention, it contains the Semen Trigonellae extract and the conventional medicine adjuvant of clinical treatment effective dose.The clinical recommendation effective dose of this medicine is 1.8g/ day.
Aforementioned pharmaceutical compositions comprises peroral dosage form, for example tablet, capsule, drop pill, pill or granule etc.
Above-mentioned preparation can prepare by conventional method.Available additive comprises excipient (for example carbohydrate derivative such as lactose, sucrose, glucose, mannitol and Sorbitol; Starch derivatives such as corn starch, potato starch, dextrin and carboxymethyl starch; Cellulose derivative such as crystalline cellulose, hydroxypropyl cellulose, carboxymethyl cellulose, carboxymethylcellulose calcium, sodium carboxymethyl cellulose; Arabic gum; Dextran; Silicate derivative such as metasilicic acid magnalium; Phosphate derivative such as calcium phosphate; Carbonate derivative such as calcium carbonate; Sulfate-derivatives such as calcium sulfate etc.), binding agent (gelatin for example, ethanol, polyvinylpyrrolidone and Polyethylene Glycol), disintegrating agent (for example cellulose derivative such as sodium carboxymethyl cellulose, polyvinylpyrrolidone), lubricant (Talcum for example, magnesium stearate, spermaceti or boric acid), stabilizing agent (methyl parahydroxybenzoate, propyl p-hydroxybenzoate etc.), correctives (for example sweeting agent and spice etc.), fluidizer (micropowder silica gel), diluent and other are auxilliary with solvents (water for example, ethanol and glycerol etc.).
The present invention also provides the above-mentioned purposes of pharmaceutical composition in the medicine of preparation treatment diabetes and hyperlipidemia that contains this extract, experimental results show that Semen Trigonellae extract of the present invention also can not add any pharmaceutic adjuvant and directly be administered to the patient with extracting solution or dry back solid form.
In a preferred embodiment of the present invention, pharmaceutical composition is specially tablet, it is characterized by the Semen Trigonellae extract with 300g, add microcrystalline Cellulose 95g, L-HPC 25g, crosslinked CMC-Na 25g, magnesium stearate 4.5g, to be pressed into 1000 behind 95% alcohol granulation, tablet wraps film-coat with 7%LE.
In another preferred embodiment of the present invention, pharmaceutical composition is specially capsule, it is characterized by the Semen Trigonellae extract with 200g, adds pregelatinized Starch 80g, crosslinked CMC-Na 12g, and micropowder silica gel 3g, with 95% alcohol granulation, promptly encapsulated.
The present active substance that has blood sugar lowering, blood fat reducing in the middle of the Semen Trigonellae and improve diabetic complications that studies show that is mainly polysaccharide, total flavonoid and saponins.Polysaccharide has significant blood sugar reducing function, saponins then has significant effect for reducing fat, flavonoid can microcirculation improvement, multiple complication to diabetics has regulating action, therefore, the therapeutical effect of Semen Trigonellae is the result of the collaborative many target spot regulation and control of this three constituents, is equivalent to 1 compound preparation.
The present invention has utilized above-mentioned synergism, by extract, preparation method such as refining, enrichment, obtained containing the Semen Trigonellae extract of these three kinds of effective ingredient, and be effective site and main active with this extract, developed the medicine that contains above-mentioned three kinds of compositions, the medicine that experiment showed, this Semen Trigonellae extract and contain this extract is having significant curative effect aspect treatment diabetes and the hyperlipidemia disease.
Pharmaceutical research:
One, the hypoglycemic activity of Semen Trigonellae extract research
Semen Trigonellae extract is to the influence of rat hyperglycemia test model due to the alloxan (AX):
140 of rats behind the fasting 12h, are prepared alloxan with normal saline before test, inject through the abdominal cavity with 200mg/kg, induce into zoic model with hyperglycemia, and preparation hypertonic glucose solution, so that in time give during the hypoglycemia convulsion at the administration initial stage.After treating 3 days its glucostasis, blood sampling, centrifugal, survey blood glucose value, get the rat of fasting blood sugar more than 180mg/dl and be divided into 7 groups by the blood glucose equilibrium, every group 10, irritate stomach respectively and give Semen Trigonellae extract (140mg/kg, 70mg/kg, 35mg/kg), TANGMAIKANG KELI (1350mg/kg) and insoral (75/kg), blank and model control group are irritated the 0.5%CMC-Na (CMC-sodium carboxymethyl cellulose) that stomach is given suitable volume every day.In 4 weeks of successive administration, once a day, relatively and in last administration (the 28th day) back different time (1,2,4, blood glucose (ginseng of knowing clearly 3.1.1) was surveyed in 24h) blood sampling, compares between fasting glucose of 7 days mensuration of every administration was organized.The 7th day Semen Trigonellae extract (140mg/kg) group begins to occur obvious blood sugar reducing function after the administration, and (140mg/kg, 70mg/kg) obvious blood sugar reducing function all appears to the 14th day Semen Trigonellae extract in group, and with the administration time prolongation, blood sugar reducing function becomes clear day by day after the administration.After the Semen Trigonellae extract drug withdrawal, its blood sugar reducing function still can be kept 4 hours.The results are shown in following table 1, table 2 and table 3.
Table 1 Semen Trigonellae extract successive administration is induced the influence (X ± SD) of hyperglycemic rat blood glucose to alloxan
Group dosage number of animals blood sugar concentration
After the preceding administration of administration
mg/kg (mg/dl) 7d 14d 21d 28d
CMC - 10 75.7±6.9 76.9±8.8 78.7±10 80.2±13.3 1±12.7
CMC+AX - 10 236.5±42.1
ΔΔ 233.5±36.2
ΔΔ 232.9±36.9
ΔΔ 247.4±74.8
ΔΔ 222.6±50.3
ΔΔ
Semen Trigonellae extract 140 10 229.6 ± 39.6 193.7 ± 48.5*, 185.3 ± 49.1*, 172.3 ± 61.7**, 166.1 ± 36.2**
70 10 235.4±56.1 195.5±52.4 185.5±31.3** 177.8±73.1* 172.4±42.3**
35 10 232.1±45.5 201.6±54.1 196.8±44.4 192.4±67.5 177.5±19.2
TANGMAIKANG 1,350 10 241.2 ± 56.5 202.6 ± 44.8 191.1 ± 36.1* 198.5 ± 74.1 176.8 ± 39.3*
Insoral 75 10 230.9 ± 37.8 184.6 ± 30.7**, 168.9 ± 33.8**, 156.5 ± 32.6**, 135.5 ± 32.9**
ΔP＜0.05,
The Δ ΔP＜0.01 contrast CMC, * P＜0.05, * * P＜0.01 contrast CMC+AX
Alloxan is induced around the table 2 Semen Trigonellae extract successive administration time-effect relationship (X ± SD) of hyperglycemic rat blood sugar influence
Dosage number of animals blood sugar concentration (mg/dl)
Group
Mg/kg 1h 2h 4h 24h
CMC - 10 71.1±12.7 76.2±11.7 92.4±13.4 106.4±12.7
CMC+AX - 10 222.6±50.3
ΔΔ 225.1±53.9
ΔΔ 229.9±53.9
ΔΔ 227.4±57.9
ΔΔ
Semen Trigonellae extract 140 10 166.1 ± 36.2** 168.3 ± 41.2* 179.5 ± 57.3* 194.8 ± 63.4
70 10 172.3±42.3* 170.0±41.1* 179.5±35.1* 197.2±43.8
35 10 177.5±59.2 181.9±51.9 186.3±40.2 202.2±65.2
TANGMAIKANG 1,350 10 176.8 ± 39.3*, 175.9 ± 36.8* 183.3 ± 58.2 190.8 ± 43.4
Insoral 75 10 135.5 ± 32.9** 156.2 ± 23.6** 162.2 ± 28.9** 203.2 ± 76.9
ΔP＜0.05,
The Δ ΔP＜0.01 contrast CMC, * P＜0.05, * * P＜0.01 contrast CMC+AX
It is as follows that administration front and back blood glucose difference is organized a paired t-test result:
Table 3 Semen Trigonellae extract successive administration is induced the influence (X ± SD) of hyperglycemic rat blood glucose to alloxan
Group dosage number of animals
mg/kg 7d 14d 21d 28d
CMC - 10 -1.2±12.3 -3.0±20.2 -4.5±16.3 4.6±21.2
CMC+AX - 10 3.0±25.6 3.6±28.7 -10.9±41.6 13.9±47.5
Semen Trigonellae extract 140 10 35.9 ± 32.1** 44.1 ± 35.6** 57.3 ± 41.1** 63.5 ± 47.9**
70 10 39.9±35.4** 50.1±39.8** 57.6±42.5** 63.1±52.1**
35 10 30.5±32.9* 35.3±36.9* 39.7±37.1** 54.6±41.9**
TANGMAIKANG 1,350 10 38.6 ± 40.1*, 50.1 ± 47.8** 42.7 ± 43.6* 64.4 ± 56.2**
Insoral 75 10 46.3 ± 41.4** 62.0 ± 47.9** 74.4 ± 56.6** 95.4 ± 68.9**
Difference compares * P＜0.05 * * P＜0.01 between organizing before and after the administration
Two, the blood fat reducing activity research of Semen Trigonellae extract
60 of mices are divided into 6 groups at random, and each administration group is given Semen Trigonellae extract (200mg/kg respectively, 100mg/kg, 50mg/kg) and zhibituo (800mg/kg), model control group and blank group are given equal-volume 0.5%CMC-Na, once a day, continuous 7 days, after the last administration 1 hour, except that the normal control group, all lumbar injection 75% egg yolk Emulsion (DH) 0.5ml/ only after 20 hours, gets blood by mice socket of the eye vein, separation of serum is surveyed cholesterol level with the T-CHOL test kit.The result show the Semen Trigonellae extract successive administration after seven days its high dose (200mg/kg) can reduce hypercholesterolemia mice serum cholesterol level.The results are shown in Table 4.
Table 4 Semen Trigonellae extract successive administration is to the influence of hypercholesterolemia mice serum cholesterol level (X ± SD)
Group dosage number of animals cholesterol (mmol/l)
(mmg/kg)
CMC - 10 3.14±0.45
CMC+DH - 10 12.07±4.27
ΔΔ
Semen Trigonellae extract 200 10 7.95 ± 2.24*
100 10 10.07±3.18
50 10 10.60±2.15
Zhibituo 800 10 9.94 ± 0.96
ΔP＜0.05,
The Δ ΔP＜0.01 contrast CMC, * P＜0.05, * * P＜0.01 contrast CMC+DH
Three, the biological activity of Semen Trigonellae extract
Semen Trigonellae extract has been carried out following pharmacodynamic study, being subjected to the reagent thing is Semen Trigonellae extract, control drug is glyburide, insoral, TANGMAIKANG (blood sugar lowering model) and zhibituo (blood fat reducing model), and pilot project is followed successively by: Semen Trigonellae extract is to the influence of normal mouse blood sugar; Semen Trigonellae extract causes the influence of rabbit, mice hyperglycemia model blood glucose to glucocorticoid; Semen Trigonellae extract causes the influence of rat hyperglycemia model blood glucose to alloxan; Semen Trigonellae extract is to being tried the influence of mice blood fat; Semen Trigonellae extract is to the influence of type ii diabetes rat model blood fat.
Result of the test shows that giving normal mouse Semen Trigonellae extract (dosage is 200mg/kg, 100mg/kg, 50mg/kg) does not have influence to mouse blood sugar, the then remarkable blood sugar lowering of control drug glyburide, blood glucose to the intact animal does not exert an influence, and is one of main feature of Semen Trigonellae extract.
Give glucocorticoid and cause hyperglycemia rabbit Semen Trigonellae extract (80mg/kg, 40mg/kg, 20mg/kg, 10mg/kg), the obvious rabbit transience blood sugar increasing that causes of antagonism exogenous glucose, middle high dose Semen Trigonellae extract (80mg/kg, 40mg/kg) can obviously promote rabbit to give sugar back serum secretion of insulin.Give glucocorticoid and cause hyperglycemia mice Semen Trigonellae extract, under high dose (200mg/kg) condition, can improve the plain opposing of mouse islets that hormone brings out.
Alloxan is caused the rat hyperglycemia model, give Semen Trigonellae extract, dosage is 140mg/kg, 70mg/kg, can significantly reduce the rat fasting blood-glucose value, obviously reduce the of short duration rising of blood glucose that the type ii diabetes rat causes owing to exogenous glucose, improve the sugared toleration of animal subject.
Semen Trigonellae extract (200mg/kg) can reduce hypercholesterolemia mice serum cholesterol level; Can reduce the serum triglycerides of type ii diabetes rat, but high density lipoprotein is not exerted an influence.
The specific embodiment
Embodiment 1: the preparation of Semen Trigonellae extract
Get dry Semen Trigonellae medical material, pulverize, cross sieve No. 2; According to the percolation under fluid extract and the extractum item (13 pages of appendix of Chinese Pharmacopoeia version in 2000), be extraction solvent with 60% ethanol, the dipping, check transudate with trichloroacetic acid, percolation to transudate to the trichloroacetic acid reaction negative till; Percolation filtrate concentrate under reduced pressure at low temperature to relative density is about 1.15-1.20; With the ethyl acetate defat; The water-soluble portion spray drying obtains Semen Trigonellae extract, after measured, contains polysaccharide 40% in this extract, total flavones 10%, saponin 20%.
Embodiment 2: the prescription and proportioning and the method for making thereof that contain the tablet of Semen Trigonellae extract
Prescription:
Semen Trigonellae extract 300g
Microcrystalline Cellulose 95g
Low-substituted hydroxypropyl cellulose 25g
95% ethanol q.s.
Cross-linking sodium carboxymethyl cellulose 25g
Magnesium stearate 4.5g
Make 1000
According to above-mentioned preparation prescription, get Semen Trigonellae extract, add microcrystalline Cellulose, the low hydroxy methocel (L-HPC) that replaces, as binding agent, cross 20 mesh sieve system soft materials, 60 ℃ of dryings with 95% ethanol, granulate adds crosslinked CMC-Na (sodium carboxymethyl cellulose) and magnesium stearate mixing tabletting; LE film coating with 7% promptly gets (LE: the product code name of Tianjin Ai Leyi medical packaging company limited).
Embodiment 3: the capsular prescription of Semen Trigonellae extract and proportioning and method for making thereof
Prescription:
Semen Trigonellae extract 200g
Pregelatinized Starch 80g
75% ethanol q.s.
Cross-linking sodium carboxymethyl cellulose 12g
Micropowder silica gel 3g
Make 1000
Get Semen Trigonellae extract, add pregelatinized Starch, as binding agent, cross 20 mesh sieve system soft materials, 60 ℃ of dryings, granulate with 95% ethanol; Dried granule added crosslinked CMC-Na of 100 purposes and micropowder silica gel mixing, measured index component content in the granule, and was encapsulated, the about 0.3g of loading amount; Packing promptly.
Embodiment 4: the prescription of Semen Trigonellae extract granule and proportioning and method for making thereof
Prescription:
Semen Trigonellae extract 1000g
Pregelatinized Starch 300g
Low-substituted hydroxypropyl cellulose 200g
95% ethanol q.s.
Make 1000 bags
Get Semen Trigonellae extract, add pregelatinized Starch and low-substituted hydroxypropyl cellulose, as binding agent, cross 20 mesh sieve system soft materials, 60 ℃ of dryings, granulate with 95% ethanol; Pack, the about 1.5g of loading amount; Packing promptly.
More than described the preferred embodiment for the present invention, so it is not in order to limit the present invention.Those skilled in the art can not depart from the improvement and the variation of category of the present invention and spirit to embodiment disclosed herein.
Claims (10)
1, the extract of a kind of Semen Trigonellae is characterized in that containing in this extract polysaccharide, the total flavones of 5-15% and the saponin of 10-30% that percentage by weight is respectively 15-50%.
2, extract as claimed in claim 1 wherein contains percentage by weight and is respectively 40% polysaccharide, 10% total flavones and 20% saponin in this extract.
3, extract as claimed in claim 1 or 2 is characterized in that, the sign composition of described total flavones is an orientin, and orientin content is 0.2-1.2% in this extract.
4, the preparation method of the described Semen Trigonellae extract of claim 1 is characterized in that, after the Semen Trigonellae pulverizing medicinal materials, extracts with pure water mixed solvent, and wherein determining alcohol is 20%-90%, and the extracting solution that obtains gets through enrichment again.
5, preparation method as claimed in claim 4, enrichment method wherein comprise that with extracting solution ethyl acetate extraction defat, the consumption of this ethyl acetate is at least 1/4 of extracting liquid volume.
6, a kind of pharmaceutical composition, it contains the Semen Trigonellae extract and the pharmacy acceptable auxiliary for the treatment of effective dose, and wherein said Semen Trigonellae extract is the mixture that contains the saponin of the total flavones of polysaccharide that percentage by weight is respectively 15-50%, 5-15% and 10-30%.
7, compositions as claimed in claim 6, it is a peroral dosage form, comprises tablet, capsule and granule, contains the Semen Trigonellae extract and the pharmacy acceptable auxiliary for the treatment of effective dose.
8, compositions as claimed in claim 7, described adjuvant is selected from microcrystalline Cellulose, low-substituted hydroxypropyl cellulose, cross-linking sodium carboxymethyl cellulose, pregelatinized Starch, micropowder silica gel and the magnesium stearate at least a.
9, the application of the described extract of claim 1 in the medicine of preparation treatment diabetes and hyperlipidemia.
10, the application of the described pharmaceutical composition of claim 4 in treatment diabetes and hyperlipidemia.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100360145C (en) * | 2005-08-25 | 2008-01-09 | 吉林大学 | Fenugreek extract and its preparation method and application |
| CN103304605A (en) * | 2013-04-27 | 2013-09-18 | 中国科学院西北高原生物研究所 | Method for preparing flavonoid glycoside and stibene glucoside type compound by separating from fenugreek |
| CN103505498A (en) * | 2012-06-29 | 2014-01-15 | 北京星昊医药股份有限公司 | Preparation and application of effective part of total flavonoids in fenugreek |
| CN105726624A (en) * | 2016-05-04 | 2016-07-06 | 吉林大学 | Pharmaceutical composition for treating diabetes |
| CN117243968A (en) * | 2023-11-13 | 2023-12-19 | 济南舜景医药科技有限公司 | Celecoxib compound preparation and preparation method thereof |
-
2004
- 2004-07-30 CN CNB2004100701046A patent/CN100434091C/en not_active Expired - Lifetime
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100360145C (en) * | 2005-08-25 | 2008-01-09 | 吉林大学 | Fenugreek extract and its preparation method and application |
| CN103505498A (en) * | 2012-06-29 | 2014-01-15 | 北京星昊医药股份有限公司 | Preparation and application of effective part of total flavonoids in fenugreek |
| CN103304605A (en) * | 2013-04-27 | 2013-09-18 | 中国科学院西北高原生物研究所 | Method for preparing flavonoid glycoside and stibene glucoside type compound by separating from fenugreek |
| CN103304605B (en) * | 2013-04-27 | 2015-04-08 | 中国科学院西北高原生物研究所 | Method for preparing flavonoid glycoside and stibene glucoside type compound by separating from fenugreek |
| CN105726624A (en) * | 2016-05-04 | 2016-07-06 | 吉林大学 | Pharmaceutical composition for treating diabetes |
| CN117243968A (en) * | 2023-11-13 | 2023-12-19 | 济南舜景医药科技有限公司 | Celecoxib compound preparation and preparation method thereof |
| CN117243968B (en) * | 2023-11-13 | 2024-02-23 | 济南舜景医药科技有限公司 | Celecoxib compound preparation and preparation method thereof |
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