CN101120954A - Medicinal composition containing GTF active extract and its preparation technology - Google Patents
Medicinal composition containing GTF active extract and its preparation technology Download PDFInfo
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- CN101120954A CN101120954A CNA2006101042774A CN200610104277A CN101120954A CN 101120954 A CN101120954 A CN 101120954A CN A2006101042774 A CNA2006101042774 A CN A2006101042774A CN 200610104277 A CN200610104277 A CN 200610104277A CN 101120954 A CN101120954 A CN 101120954A
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Abstract
The present invention discloses a medical compound containing the GTF active extracts and the production art as well as the new function of adjusting the leptin gene and the leptin receptor gene expression. The material of the compound in the present invention comprises the GTF active extracts and the chitin. The medicine of the present invention is applied in preparation of medicine for treatment of the insufficient glucose tolerance (IGT) and the hyperlipidemia.
Description
Technical field
The present invention relates to the new purposes of a kind of pharmaceutical composition and preparation technology and effect thereof, particularly a kind of pharmaceutical composition and preparation technology thereof who contains GTF activity extract (glucose tolerance factor activity extract).
Background technology
The GTF activity extract is a kind of active component that promotes the body glucose utilization that has that makes of separating from the special beer yeast.Studies show that in a large number up to now, the GTF activity extract is having important effect aspect the sugar of regulating body and the lipid metabolism, has potential clinical value for the treatment of diabetes and hyperlipemia.This research is on the basis of these researchs, further improve the extraction process of GTF activity extract, and adopted the GTF active extractive combination of other being combined to form property of active polymeric substance, be used for the control of carbohydrate tolerance low (IGT), diabetes and hyperlipemia.
Summary of the invention
One object of the present invention is to disclose a kind of GTF of containing activity extract pharmaceutical composition; Another object of the present invention is to disclose a kind of extracting method of GTF activity extract; The 3rd purpose of the present invention is to disclose this preparation of drug combination technology; The 4th purpose of the present invention is to disclose the new purposes of this pharmaceutical composition.
The present invention seeks to be achieved through the following technical solutions:
The raw material of pharmaceutical composition of the present invention consists of: GTF activity extract 10-50 weight portion, chitin 30-60 weight portion, dextrin 20-35 weight portion.
The raw material composition of pharmaceutical composition of the present invention is preferably: GTF activity extract 30-47 weight portion, chitin 33-43 weight portion, dextrin: 20-27 weight portion.
The crude drug composition of pharmaceutical composition of the present invention is preferably: GTF activity extract 47 weight portions, chitin 33 weight portions, dextrin 20 weight portions.
The raw material composition of pharmaceutical composition of the present invention is preferably: GTF activity extract 24-34 weight portion, chitin 41-47 weight portion, dextrin 25-29 weight portion.
The crude drug composition of pharmaceutical composition of the present invention is preferably: GTF activity extract 35 weight portions, chitin 40 weight portions, dextrin 25 weight portions.
The raw material composition of pharmaceutical composition of the present invention is preferably: GTF activity extract 12-17 weight portion, chitin 53-55 weight portion, dextrin 30-33 weight portion.
The raw material composition of pharmaceutical composition of the present invention is preferably: GTF activity extract 17 weight portions, chitin 53 weight portions, dextrin 30 weight portions.
Get the aforementioned pharmaceutical compositions raw material, according to common process, make clinical acceptance but be not limited to preparations such as tablet, granule, capsule, oral liquid, drop pill, injection.
Wherein said GTF activity extract can prepare according to following process: take by weighing 20-40 weight portion weak-base ion-exchange resin (available DEAE-11, DEAE-22, DEAE-23, DEAE-32 or DEAE51-53 model are replaced) dry powder, join earlier in the 0.2-1NNaOH solution of 130-170 parts by volume, the limit edged stirs, place and soak after 0.5-1.0 hour in the beaker, use the distilled water eluting, be washed till neutrality, join again in the 0.2-2NHCL solution of 130-170 parts by volume, be washed till neutrality with quadrat method; Get strong acidic ion resin (available Dowex50W-X2, X4, X8 or X12 model are replaced) dry powder 20-40 weight portion, processing method is with weak-base ion-exchange resin dry powder;
Take by weighing the Rich chromium yeast powder of 20-40 weight portion, dose volume adds yeast powder in the above-mentioned mixed liquor than being the n-butanol-water mixed liquor 400-800 parts by volume of 0.5-1.5: 0.5-1.5, stirs 2-4 hour, left standstill 12-20 hour, the supernatant of getting aqueous phase is at 50 ℃ of-60 ℃ of following vacuum concentration; The distilled water that concentrated solution is added 2-4 times of parts by volume is dialysed, and merges dialysis solution, vacuum concentration; Dialysis solution after will concentrating adds to be crossed post and separates in the weak-base ion-exchange resin chromatographic column, and use the distilled water eluting, collect eluent up to ultraviolet absorption peak till 262nm place does not have absorption, concentrate eluant; Concentrated solution is separated through the strong acidic ion resin post, use 0.25M NH
4OH eluting, collection eluent till the 262nm place does not absorb, are concentrated into residue 10-30 parts by volume with this eluent up to ultraviolet absorption peak, and lyophilization obtains the GTF activity extract.
Above-mentioned described weak-base ion-exchange resin is meant: DEAE cellulose-23 (DEAE-cellulose), and also can select DEAE-11, DEAE-22, DEAE-23, DEAE-32 or DEAE51-53 model to replace; Wherein strong acidic ion resin is meant: Dowex50WX8 (732 strong acidic ion resin), also can select Dowex50W-X2, X4, X8 or X12 model to replace.
Above-mentioned described cryodesiccated freezing conditions is: condition is-40 ℃ to-35 ℃, 12-15Pa, 28-32h.
The preparation technology of GTF activity extract is preferably: take by weighing 30 weight portion DEAE-23 celluosic resin dry powder, join in the 0.2NNaOH solution of about 150 parts by volume, the limit edged stirs, place and soak after 0.6 hour in the beaker, use the distilled water eluting, till washing neutrality, join again in the 0.2NHCL solution of 150 parts by volume, be washed till neutrality with quadrat method; Get Dowex50WX8 strong acidic ion resin dry powder 20 weight portions, processing method is with DEAE-23 celluosic resin dry powder;
Take by weighing the Rich chromium yeast powder of 30 weight portions, the dose volume ratio is 1: 1 n-butanol-water mixture 600 parts by volume, and yeast powder is added in the above-mentioned mixed liquor, stirs 3 hours, leaves standstill 20h, and the supernatant of getting aqueous phase is at 55 ℃ of following vacuum concentration; The distilled water that concentrated solution is added 2 times of parts by volume is respectively dialysed, and merges dialysis solution, vacuum concentration; Dialysis solution added in the DEAE-23 celluosic resin chromatography post crosses post and separate, use the distilled water eluting, collect eluent up to ultraviolet absorption peak till 262nm place does not have absorption, concentrate eluant; Concentrated solution is separated through the Dowex50WX8 cationic resin column, use 0.25MNH
4The OH eluting, collect eluent up to ultraviolet absorption peak at 262nm place not till the absorption, this eluent is concentrated into residue 20 parts by volume, put into the lyophilization of vacuum lyophilization instrument, condition is-40 ℃ to-35 ℃, 12-15Pa, 28-32h, obtains the sticking slightly GTF activity extract of yellowish-brown.
GTF activity extract of the present invention is meant glucose tolerance factor, and wherein Huo Chrome content is not less than 1360mg/Kg.
The ratio of weight portion of the present invention and parts by volume is g/ml.
Pharmaceutical composition of the present invention is used for the medicine of preparation treatment carbohydrate tolerance low (IGT), diabetes or hyperlipemia.
Description of drawings:
Fig. 1: DEAE-23 cellulose column water elution figure (along with the increase of eluting bottle number, light absorption value descends thereupon)
Fig. 2: Dowex50WX8 resin ammonia elution profile (along with the increase of eluting bottle number, light absorption value descends thereupon)
Fig. 3: the infrared spectrogram of GTF activity extract
Pharmaceutical composition of the present invention is proved through clinical trial, has the glucose utilization that promotes yeast cells; Regulate blood pressure and blood lipoid; Raise test-type diabetes rat insulin receptor gene and express, improve sugar tolerance; The obesity of effectively preventing high fat diet to induce; The leptin level that adjusting is induced by high fat diet and the effect of OB-RmRNA thereof.
Following experiment and embodiment are used for further specifying but are not limited to the present invention.
The bioactive mensuration of experimental example 1 GTF
This experiment adopts the brewer's yeast cultivation that the activity of extracting the pharmaceutical composition group (embodiment of the invention 1 Capsule content) of gained GTF activity extract (embodiment of the invention 7 is obtained) and GTF activity extract by technical solution of the present invention is measured, and now is reported as follows:
The cultivation of 1 bacterial classification: configuration culture medium: glucose 60g, dusty yeast 10g, (NH4)
2SO
410g、
MgSO
41g、KH
2PO
42g、K
2HPO
42g. Each 200ml of packing, autoclaving 20min is inoculated in brewer's yeast in the fluid nutrient medium, puts shaking table cultivation 48h (30 ℃) in the incubator
2 preparation yeast resting cells: will cultivate gained centrifugal (4000r/min, 15min), abandoning supernatant, again centrifugal with aqua sterilisa flushing one time, obtain the yeast thalline. Be inoculated in respectively and cultivate 48h (30 ℃) in the mid-shaking table of aqua sterilisa, centrifugal, abandoning supernatant, with the aqua sterilisa flushing, centrifugal. Repeat again this step, obtain the yeast resting cell.
3 application of samples: get the 500ml conical flask, every packing 200ml distilled water adds respectively 6g glucose, autoclaving 20min. Inoculation 5g yeast resting cell in every conical flask. Above-mentioned conical flask is divided into four groups: it is 70 μ g/ml that the pharmaceutical composition group of control group (not adding any sample), Cr-enriched yeast powder group, GTF activity extract group and GTF activity extract all contains chromium ion concentration), all place shaking table to cultivate (30 ℃).
4 sampling and measuring: respectively at cultivating 4h, 6h, 7h, 8h, 9h, 10h, the 11h 5ml that takes a sample, centrifugal (4000r/min, 15min) gets supernatant, survey wherein remaining concentration of glucose with glucose oxidase method, be used for reacting not on the same group between the degree of glucose metabolism.
(seeing Table 1)
The comparison of remaining concentration of glucose in the table 1 different time reaction system
| 4h | 6h | 7h | 8h | 9h | 10h | 11h | |
| Reference substance (blank) Cr-enriched yeast powder GTF activity extract GTF pharmaceutical composition | 88.37±0.56 87.73±1.36 88.19± 0.82 88.54± 0.54 | 78.58±1.64 80.36±1.35 76.27± 1.40 ***△△△ 73.86± 2.12 ***△△△# | 73.59±1.73 73.32±1.34 68.45± 1.87 ***△△△ 66.34± 1.22 ***△△△# | 74.75±1.85 65.56±1.46 49.57± 1.56 ***△△△ 47.18± 1.88 ***△△△# | 43.88±1.23 50.12±1.59 28.09± 1.49 ***△△△ 26.04± 1.38 ***△△△# | 33.82±1.67 34.34±1.89 13.95± 1.35 ***△△△ 13.15± 1.25 ***△△△# | 21.65±1.38 24.42±1.59 6.55± 0.89 ***△△△ 2.43± 0.45 ***△△△# |
Compare * P＜0.05 with the blank group, * * P＜0.01, △ P＜0.05 is compared with Cr-enriched yeast powder group in * * * P＜0.001, △ △ P＜0.01, △ △ △ P＜0.001, composition and GTF group be #P＜0.05 relatively
Experimental result shows, the pharmaceutical composition of the GTF activity extract that is made by this experiment and the effect that the GTF activity extract all has significant promotion yeast cells glucose utilization, compare with blank group, Cr-enriched yeast powder group, after the 6th hour, remaining concentration of glucose all has significant difference (P＜0.001) in the reaction system. Pharmaceutical composition group and the GTF activity extract group of GTF activity extract more variant (P＜0.005).
The pharmaceutical composition group of experimental example 2GTF activity extract is on the impact of Experimental Mice fasting blood-glucose and sugar tolerance
The alloxan diabetes mouse is divided into model group, model adds GTF activity extract group (embodiment of the invention 8 is obtained), model adds the chitin group, model add the pharmaceutical composition group (embodiment of the invention 2 is obtained) that contains the GTF activity extract little, in, heavy dose of (be equivalent to clinical dosage 5,10,20 times), model adds glibenclamide group (glibenclamide 5mg/kg), Normal group and model control group gavage give equal-volume distilled water, every day 1 time, continuous 10 days. Carry out respectively the mensuration of blood sugar and sugar tolerance
The pharmaceutical composition of table 2GTF activity extract is on the impact of alloxan mouse fasting blood-glucose
| Group | n | Blood glucose value | |
| 0 day | 11 days | ||
| The heavy dose of group of dosage group GTF pharmaceutical composition in the Normal group model group positive drug group GTF activity extract group chitin group GTF pharmaceutical composition small dose group GTF pharmaceutical composition | 10 10 10 10 10 10 10 10 | 4.83±0.75 21.89±3.40 *** 21.79±3.20 *** 21.47±3.56 *** 21.59±3.9 *** 21.03±3.26 *** 21.12±3.51 *** 21.59±3.71 *** | 4.72±0.73 18.62±3.43 *** 9.82+3.0 △△△ 16.35±2.38 △△ 16.89±2.67 △△ 15.62±2.59 △△ 14.88±2.30 △△ 11.62±2.98 △△ |
Compare with Normal group: * * * P＜0.001; Compare with model group: △ △ P＜0.001, △ △ △ P＜0.001
The pharmaceutical composition of table 3GTF activity extract is on the impact of alloxan glucose tolerance in mice
| Group | n | Blood glucose value | |||
| 0min | 30min | 60min | 120min | ||
| The heavy dose of group of dosage group GTF pharmaceutical composition in the Normal group model group GTF activity extract group chitin group GTF pharmaceutical composition small dose group GTF pharmaceutical composition | 10 10 10 10 10 10 10 | 5.16±0.97 16.76±2.88 *** 14.26±2.69 △△ 15.60±2.78 △ 12.03±2.36 △△△ 12.90±2.50 △△ 11.60±2.03 △△△ | 5.74±0.92 20.03±3.25 *** 16.38±2.59 △△ 17.69±2.68 △ 14.78±2.16 △△ 15.12±3.09 △△ 14.36±1.85 △△△ | 5.27±0.82 17.37±2.55 *** 15.49±2.37 △△ 16.31±2.29 △ 13.35±2.89 △△ 13.21±3.15 △△ 12.04±2.61 △△△ | 5.29±0.62 14.58±2.92 *** 12.78±2.35 △△ 13.36±2.95 △ 11.27±2.30 △△ 11.09±2.29 △△ 11.36±1.96 △△ |
Compare with the normal control group: * * * P＜0.001; Compare with model group: △ △ △ P＜0.001, △ △ P＜0.01, △ P＜0.05
Experiment finds that the pharmaceutical composition group, GTF activity extract group, the chitin group that contain the GTF activity extract all can reduce alloxan mice fasting glucose content, relatively have significant difference (P＜0.01 or P＜0.001) with model group.Can significantly improve simultaneously the carbohydrate tolerance level of diabetic mice, with model group significant difference (P＜0.01 or P＜0.001) is arranged relatively, the blood glucose value of the pharmaceutical composition group of GTF activity extract significantly is lower than GTF activity extract group, chitin group (P＜0.05, P＜0.01).
The influence that the pharmaceutical composition group of experimental example 3GTF activity extract is expressed experimental mouse liver insulin receptor gene
The alloxan diabetes mice is divided into model group, model adds the pharmaceutical composition group that contains the GTF activity extract, and ((embodiment of the invention 5 is obtained, be equivalent to 20 times of clinical dosages), model adds GTF activity extract group ((embodiment of the invention 9 is obtained) is equivalent to 20 times of clinical dosages), model adds chitin group (being equivalent to 20 times of clinical dosages), model adds glyburide group (being equivalent to 20 times of clinical dosages), normal control group and model control group are irritated stomach and are given the equal-volume distilled water, every day 1 time, after continuous 10 days, take out the animal livers tissue rapidly, adopt the RT-PCR method to detect the situation that insulin receptor gene is expressed.
The influence that the pharmaceutical composition group of table 4GTF activity extract is expressed experimental mouse liver insulin receptor gene
| The crest volume | The blank group | The glyburide matched group | The GTF pharmaceutical composition | The GTF activity extract | The chitin group | Normal group |
| β-actin (1) insulin receptor gene (2) (2)/(1) | 54761 35648 0.65 | 59106 38817 0.66 | 56511 39137 0.69 | 57634 39191 0.68 | 58349 38510 0.66 | 56785 41548 0.73 |
Table 4 shows, through PT-PCR, compare with matched group behind the GDS-8000 densitometric scan, contain the drug regimen group of GTF activity extract and the GTF activity extract group impaired insulin receptor gene mRNA level of diabetic animal that is significantly improved, and the drug regimen group that contains the GTF activity extract is better than GTF activity extract group and glyburide group.
The pharmaceutical composition group of experimental example 4GTF activity extract is to the influence of obese rat body weight, blood fat and insulin, leptin level
After adaptability was fed a week after animal was purchased, wherein 12 were adopted normal diets, and other 84 are adopted high lipid foods, make the obese rat model.(the normal group body weight is 283.75 ± 23.37 in the modeling success, fat model group body weight is 377.17 ± 21.52) the obese rat weight average be normal rats body weight 1.3-1.4 doubly) after, experimental obese rat is divided into 7 groups at random, every group 12: model group, positive drug (Xuezhikang) group, the pharmaceutical composition group of GTF activity extract (embodiment of the invention 3 is obtained) small dose group, middle dosage group, heavy dose of group (5 times of the clinical dosage that is equivalent to respectively to be grown up, 10 times, 20 times), GTF activity extract group (be equivalent to be grown up clinical dosage 10 times), chitin group (be equivalent to be grown up clinical dosage 10 times) beginning gastric infusion, normal group and model group are irritated stomach and are given normal saline.Feed with the normal diet except that normal group simultaneously, all the other groups are still fed high lipid food, in continuous 4 weeks, survey body weight weekly, fasting 12h after the last administration, morning next day, open the breast heart extracting blood under the etherization, standing separation serum ,-20 ℃ of preservations are to be measured, enzymatic assays serum TC, TG, HDL-C, LDL-C, the content of serum measured by radioimmunoassay insulin, leptin.Open cranium rapidly and isolate hypothalamus, place Trizol ,-20 ℃ of preservations are to be measured.Detect the situation that the hypothalamus leptin receptor gene is expressed with fluorescence quantifying PCR method.
(1) the pharmaceutical composition group of GTF activity extract is to the influence (seeing Table 5) of the fat rat model body weight of diet induced
The variation of the fat rat model body weight of table 5 diet induced
| Group | Before the treatment | After the treatment | ||||
| First week | Second week | The 3rd week | Around | Increasing value | ||
| Normal group | 283.75±23.37 * | 292.92±34.21 * | 306.25±43.18 * | 305.42±34.67 * | 310.83±31.97 * | 27.08±16.3 * |
| Fat model group | 376.25±22.58 | 386.67±24.80 | 398.75±21.76 | 407.50±29.12 | 428.33±32.43 | 52.08±18.64 |
| Xuezhikang group GTF activity extract group chitin group | 375.42±22.71 378.37±20.57 379.56±18.06 | 382.92±19.71 385.78±20.61 385.08±19.37 | 388.75±21.96 389.36±20.56 397.57±21.08 | 394.17±22.85 396.77±21.34 405.68±22.08 | 405.42±18.88 * 407.67±19.56 * 427.37±22.78 | 30.00±13.31 * 29.16±11.02 * 49.37±10.89 |
| GTF pharmaceutical composition small dose group | 375.83±19.98 | 383.75±20.13 | 387.92±21.79 | 395.42±24.72 | 404.58±25.62 * | 28.75±9.32 * |
| Dosage group in the GTF pharmaceutical composition | 380.42±21.79 | 374.58±16.44 | 383.75±24.97 | 397.08±23.98 | 408.33±20.93 | 27.92±10.76 * |
| The heavy dose of group of GTF pharmaceutical composition | 377.92±23.78 | 400.00±24.40 | 400.83±22.95 | 401.67±21.25 | 406.25±23.37 * | 28.33±9.61 * |
Compare * P＜0.05 * * P＜0.01 with fat model group
Table 5 as seen, the rat body weight of fat group and 8 experimental grouies does not have difference before treatment, As time goes on, each organizes the weight increase of rat, but fat group of the growth trend of experimental group rat and chitin group are slow.When the 4th week, the pharmaceutical composition group small dose group of GTF activity extract significantly is lower than fat group (P＜005) with the body weight of heavy dose of group, and there were significant differences (P＜005) with fat group for each the dosage group of pharmaceutical composition of treatment back GTF activity extract and the increasing value of GTF activity extract group body weight.
(2) the pharmaceutical composition group of GTF activity extract is to the influence (seeing Table 6) of blood fat
The pharmaceutical composition group of table 6GTF activity extract to the influence of blood fat (X ± SD, mmol/L)
| Group | TG | TC | HDL-C | LDL-C |
| The heavy dose of group of dosage group GTF pharmaceutical composition in the fat model group Xuezhikang of the normal group group GTF activity extract group chitin group GTF pharmaceutical composition small dose group GTF pharmaceutical composition | 1.10±0.56 △ 1.43±0.82 * 1.08±0.21 △ 1.01±0.18 △△ 1.16±0.19 △ 0.94±0.17 △△ 0.84±0.14 △△△ 0.82±0.30 △△△ | 1.33±0.15 △ 1.57±0.21 * 1.44±0.11 1.40±0.18 △ 1.47±0.27 1.38±0.30 △ 1.48±0.11 1.43±0.29 | 0.52±0.14 △ 0.38±0.06 * 0.55±0.14 △△ 0.48±0.08 △ 0.45±0.15 0.50±0.15 △ 0.63±0.09 △△△ 0.50±0.13 △ | 0.43±0.20 0.55±0.21 0.47±0.22 0.45±0.18 0.47±0.15 0.43±0.17 0.49±0.17 0.40±0.26 |
Compare * P＜0.05 with normal group, * * P＜0.01, △ P＜0.05 is compared with fat model group in * * * P＜0.001, △ △ P＜0.01, △ △ △ P＜0.00
See Table 6, each dosage group of the pharmaceutical composition group of GTF activity extract, GTF activity extract group, chitin group all can significantly reduce the TG level, with model group significant difference (P＜0.01 is arranged relatively, P＜0.001), the pharmaceutical composition group small dose group of GTF activity extract and GTF activity extract group can reduce the TC level, with model group more variant (P＜0.05), middle heavy dose of group has reduction trend.Each dosage group of the pharmaceutical composition group of GTF activity extract and GTF activity extract group are to the HDL-C effect of being significantly improved, with model group more variant (P＜0.05, P＜0.001).The horizontal no significant difference of LDL-C between each group.Each dosage group of the pharmaceutical composition group of GTF activity extract all is better than GTF activity extract group, chitin group aspect blood fat reducing.
(3) the pharmaceutical composition group of GTF activity extract is to the influence (seeing Table 7) of serum insulin, leptin content
The pharmaceutical composition of table 7GTF activity extract is to the influence of serum insulin, leptin content (X ± SD)
| Group | Insulin (uIU/ml) | Leptin (ng/ml) |
| The heavy dose of group of dosage group GTF pharmaceutical composition in the fat model group Xuezhikang of the normal group group GTF activity extract chitin group GTF pharmaceutical composition small dose group GTF pharmaceutical composition | 13.21±4.74 △ 16.83±4.30 * 15.39±4.25 15.47±3.78 16.78±4.16 * 15.34±2.16 14.21±2.06 13.78±1.90 △ | 0.83±0.37 △ 1.17±0.25 * 0.99±0.40 0.96±0.35 1.18±0.37 * 0.86±0.22 △ 1.04±0.38 1.08±0.52 |
Compare * P＜0.05 with normal group, * * P＜0.01, △ P＜0.05 is compared with fat model group in * * * P＜0.001, △ △ P＜0.01, △ △ △ P＜0.001
The inductive obese rat serum insulin of high fat diet, leptin level and normal group are more variant.Table 7 shows that the pharmaceutical composition of GTF activity extract can improve obese rat hyperinsulinemia and high leptin mass formed by blood stasis.
(4) the pharmaceutical composition group of GTF activity extract is to the influence (seeing Table 8) of the fat rat model hypothalamus of diet induced OB-R gene expression
The comparison of hypothalamus O B-R gene expression dose is respectively organized in table 8 experiment
| Group | -Δ Δ Ct value |
| The heavy dose of group of dosage group GTF pharmaceutical composition in the fat model group Xuezhikang of the normal group group GTF activity extract chitin group GTF pharmaceutical composition small dose group GTF pharmaceutical composition | 0.313±0.651 ** -0.750±0.677 -0.682±0.815 -0.109±0.378 -0.698±0.534 -0.045±1.011 * -0.100±0.615 -0.300±0.587 |
Compare * P＜0.05**P＜0.01 with fat model group
Table 8 as seen, fat model group hypothalamus OB-R gene expression relative quantification value reduces, with normal group than P＜001, difference has the highly significant meaning, the pharmaceutical composition group small dose group hypothalamus OB-R gene expression relative quantification value of GTF activity extract increases, with matched group than P＜005, difference also has the significance meaning.
Following embodiment all can realize the effect of above-mentioned test example
The specific embodiment
Embodiment 1: the preparation of capsule
Get GTF activity extract 47g and mix with chitin 33g, dextrin 20g respectively, promptly obtain the dried medicated powder that 100g contains GTF activity extract active component, the medicated powder that makes is made every 0.2g of capsule according to conventional method, once a day, each one;
The preparation technology of wherein said GTF activity extract is: take by weighing 30g DEAE-23 celluosic resin (DEAE-cellulose) dry powder, join in the 0.2NNaOH solution of about 150ml, the limit edged stirs, place and soak after 0.6 hour in the beaker, use the distilled water eluting, till washing neutrality, join again to use with quadrat method in the 0.2NHCL solution of 150 parts by volume and wash neutrality; The processing of getting Dowex50WX8 resin (732 strong acidic ion resin) is with the DEAE-23 celluosic resin; Take by weighing the Rich chromium yeast powder of 30g, the dose volume ratio is 1: 1 n-butanol-water mixture 600ml, and yeast powder is added in the above-mentioned mixed liquor, stirs 3 hours, leaves standstill 20h, and the supernatant of getting aqueous phase is at 55 ℃ of following vacuum concentration; Concentrated solution is added respectively in the distilled water of 2 times of parts by volume and dialyse, merge dialysis solution, vacuum concentration; Dialysis solution added in the DEAE-23 cellulose chromatography post crosses post and separate, use the distilled water eluting, collect eluent up to ultraviolet absorption peak till 262nm place does not have absorption, concentrate eluant; Concentrated solution is separated through the Dowex50WX8 resin column, use 0.25MNH
4The OH eluting, collect eluent up to ultraviolet absorption peak at 262nm place not till the absorption, this eluent is concentrated into thick (remaining 20 parts by volume), put into the lyophilization of vacuum lyophilization instrument, condition is-40 ℃ to-35 ℃, 12-15Pa, 28-32h, obtains the sticking slightly material GTF of yellowish-brown activity extract.
Embodiment 2: the preparation of tablet
Get GTF activity extract 17g and mix with chitin 53g, dextrin 30g respectively, promptly obtain the dried medicated powder that 100g contains GTF activity extract active component, the medicated powder that makes is made tablet according to conventional method.Every 0.5g, once a day, each a slice;
The preparation technology of wherein said GTF activity extract is: take by weighing 30g DEAE-23 celluosic resin (DEAE-cellulose) dry powder, join in the 0.2NNaOH solution of about 160ml, the limit edged stirs, place and soak after 0.6 hour in the beaker, use the distilled water eluting, till washing neutrality, join again in the 0.2NHCL solution of 160 parts by volume, use with quadrat method and wash neutrality; The processing of getting Dowex50WX8 resin (732 strong acidic ion resin) is with the DEAE-23 celluosic resin; Take by weighing the Rich chromium yeast powder of 20g, the dose volume ratio is 1.: 1 n-butanol-water mixture 400ml, and yeast powder is added in the above-mentioned mixed liquor, stirs 3 hours, leaves standstill 20h, and the supernatant of getting aqueous phase is at 60 ℃ of following vacuum concentration; Concentrated solution is added respectively in the distilled water of 2 times of parts by volume and dialyse, merge dialysis solution, vacuum concentration; Dialysis solution added in the DEAE-23 cellulose chromatography post crosses post and separate, use the distilled water eluting, collect eluent up to ultraviolet absorption peak till 262nm place does not have absorption, concentrate eluant; Concentrated solution is separated through the Dowex50WX8 resin column, use 0.25MNH
4The OH eluting, collect eluent up to ultraviolet absorption peak at 262nm place not till the absorption, this eluent is concentrated into thick (remaining 10 parts by volume), put into the lyophilization of vacuum lyophilization instrument, condition is-40 ℃ to-35 ℃, 12-15Pa, 28-32h, obtains the sticking slightly material GTF of yellowish-brown activity extract.
Embodiment 3: the preparation of oral liquid
Get GTF activity extract 0.9g, be dissolved in the 91ml pure water, make oral liquid according to conventional method, every 10ml, once a day, each one;
The preparation technology of wherein said GTF activity extract is: take by weighing 30g DEAE-23 celluosic resin (DEAE-cellulose) dry powder, join among the 0.2NNaOH and 0.2NHCL solution of about 140ml, the limit edged stirs, place and soak after 0.5 hour in the beaker, use the distilled water eluting, till washing neutrality, join again in the 0.2NHCL solution of 140 parts by volume, use with quadrat method and wash neutrality; The processing of getting Dowex50WX8 resin (732 strong acidic ion resin) is with the DEAE-23 celluosic resin; Take by weighing the Rich chromium yeast powder of 40g, the dose volume ratio is 1: 1 n-butanol-water mixture 800ml, and yeast powder is added in the above-mentioned mixed liquor, stirs 4 hours, leaves standstill 20h, and the supernatant of getting aqueous phase is at 50 ℃ of following vacuum concentration; Concentrated solution is added respectively in the distilled water of 2 times of parts by volume and dialyse, merge dialysis solution, vacuum concentration; Dialysis solution added in the DEAE-23 cellulose chromatography post crosses post and separate, use the distilled water eluting, collect eluent up to ultraviolet absorption peak till 262nm place does not have absorption, concentrate eluant; Concentrated solution is separated through the Dowex50WX8 resin column, use 0.25MNH
4The OH eluting, collect eluent up to ultraviolet absorption peak at 262nm place not till the absorption, this eluent is concentrated into thick (remaining 30 parts by volume), put into the lyophilization of vacuum lyophilization instrument, condition is-40 ℃ to-35 ℃, 12-15Pa, 28-32h, obtains the sticking slightly material GTF of yellowish-brown activity extract.
Embodiment 4: the preparation of drop pill
Get GTF activity extract 47g and mix with chitin 33g, dextrin 20g respectively, promptly obtain the dried medicated powder that 100g contains GTF activity extract active component, the medicated powder that makes is made drop pill according to conventional method;
The preparation technology of wherein said GTF activity extract is: take by weighing 30g DEAE-23 celluosic resin (DEAE-cellulose, available DEAE-11, DEAE-22, DEAE-23, DEAE-32, DEAE51-53 model are replaced) dry powder, join in the 0.2NNaOH solution of about 150ml, the limit edged stirs, place in the beaker and soak after 0.6 hour, use the distilled water eluting, till washing neutrality, join again in the 0.2NHCL solution of 150 parts by volume, use with quadrat method and wash neutrality; The processing of getting Dowex50WX8 resin (732 strong acidic ion resins, available Dowex50W-X2, X4, X8, X12 model are replaced) is with the DEAE-23 celluosic resin; Take by weighing the Rich chromium yeast powder of 30g, the dose volume ratio is 1: 1 n-butanol-water mixture 600ml, and yeast powder is added in the above-mentioned mixed liquor, stirs 3 hours, leaves standstill 20h, and the supernatant of getting aqueous phase is at 55 ℃ of following vacuum concentration; Concentrated solution is added respectively in the distilled water of 2 times of parts by volume and dialyse, merge dialysis solution, vacuum concentration; Dialysis solution added in the DEAE-23 cellulose chromatography post crosses post and separate, use the distilled water eluting, collect eluent up to ultraviolet absorption peak till 262nm place does not have absorption, concentrate eluant; Concentrated solution is separated through the Dowex50WX8 resin column, use 0.25MNH
4The OH eluting, collect eluent up to ultraviolet absorption peak at 262nm place not till the absorption, this eluent is concentrated into thick (remaining 20 parts by volume), put into the lyophilization of vacuum lyophilization instrument, condition is-40 ℃ to-35 ℃, 12-15Pa, 28-32h, obtains the sticking slightly material GTF of yellowish-brown activity extract.
Embodiment 5: the preparation of granule
Get GTF activity extract 25g and mix with chitin 45g, dextrin 25g respectively, promptly obtain the dried medicated powder that 95g contains GTF activity extract active component, the medicated powder that makes is made granule according to conventional method;
The preparation technology of wherein said GTF activity extract is: take by weighing 30g DEAE-23 celluosic resin (DEAE-cellulose, available DEAE-11, DEAE-22, DEAE-23, DEAE-32, DEAE51-53 model are replaced) dry powder, join in the 0.2NNaOH solution of about 140ml, the limit edged stirs, place in the beaker and soak after 0.8 hour, use the distilled water eluting, till washing neutrality, join again in the 0.2NHCL solution of 140 parts by volume, use with quadrat method and wash neutrality; The processing of getting Dowex50WX8 resin (732 strong acidic ion resins, available Dowex50W-X2, X4, X8, X12 model are replaced) is with the DEAE-23 celluosic resin; Take by weighing the Rich chromium yeast powder of 20g, the dose volume ratio is 1: 1 n-butanol-water mixture 400ml, and yeast powder is added in the above-mentioned mixed liquor, stirs 3 hours, leaves standstill 20h, and the supernatant of getting aqueous phase is at 60 ℃ of following vacuum concentration; Concentrated solution is added respectively in the distilled water of 2 times of parts by volume and dialyse, merge dialysis solution, vacuum concentration; Dialysis solution added in the DEAE-23 cellulose chromatography post crosses post and separate, use the distilled water eluting, collect eluent up to ultraviolet absorption peak till 262nm place does not have absorption, concentrate eluant; Concentrated solution is separated through the Dowex50WX8 resin column, use 0.25MNH
4The OH eluting, collect eluent up to ultraviolet absorption peak at 262nm place not till the absorption, this eluent is concentrated into thick (remaining 10 parts by volume), put into the lyophilization of vacuum lyophilization instrument, condition is-40 ℃ to-35 ℃, 12-15Pa, 28-32h, obtains the sticking slightly material GTF of yellowish-brown activity extract.
Embodiment 6: the preparation of injection
Get GTF activity extract 0.9g, be dissolved in the 91ml pure water, the medicated powder that makes is made injection according to conventional method;
The preparation technology of wherein said GTF activity extract is: take by weighing 30g DEAE-23 celluosic resin (DEAE-cellulose) dry powder, join in the 0.2NNaOH solution of about 160ml, the limit edged stirs, place and soak after 0.5 hour in the beaker, use the distilled water eluting, till washing neutrality, join again in the 0.2NHCL solution of 160 parts by volume, use with quadrat method and wash neutrality; The processing of getting Dowex50WX8 resin (732 strong acidic ion resin) is with the DEAE-23 celluosic resin; Take by weighing the Rich chromium yeast powder of 40g, the dose volume ratio is 1: 1 n-butanol-water mixture 800ml, and yeast powder is added in the above-mentioned mixed liquor, stirs 4 hours, leaves standstill 20h, and the supernatant of getting aqueous phase is at 50 ℃ of following vacuum concentration; Concentrated solution is added respectively in the distilled water of 2 times of parts by volume and dialyse, merge dialysis solution, vacuum concentration; Dialysis solution added in the DEAE-23 cellulose chromatography post crosses post and separate, use the distilled water eluting, collect eluent up to ultraviolet absorption peak till 262nm place does not have absorption, concentrate eluant; Concentrated solution is separated through the Dowex50WX8 resin column, use 0.25MNH
4The OH eluting, collect eluent up to ultraviolet absorption peak at 262nm place not till the absorption, this eluent is concentrated into thick (remaining 30 parts by volume), put into the lyophilization of vacuum lyophilization instrument, condition is-40 ℃ to-35 ℃, 12-15Pa, 28-32h, obtains the sticking slightly material GTF of yellowish-brown activity extract.
The preparation of embodiment 7:GTF activity extract
The preparation technology of GTF activity extract is: take by weighing 30g DEAE-23 celluosic resin (DEAE-cellulose) dry powder, join in the 0.2NNaOH solution of about 150ml, the limit edged stirs, place and soak after 0.6 hour in the beaker, use the distilled water eluting, till washing neutrality, join again in the 0.2NHCL solution of 150 parts by volume, use with quadrat method and wash neutrality; The processing of getting Dowex50WX8 resin (732 strong acidic ion resin) is with the DEAE-23 celluosic resin; Take by weighing the Rich chromium yeast powder of 30g, the dose volume ratio is 1: 1 n-butanol-water mixture 600ml, and yeast powder is added in the above-mentioned mixed liquor, stirs 3 hours, leaves standstill 20h, and the supernatant of getting aqueous phase is at 55 ℃ of following vacuum concentration; Concentrated solution is added respectively in the distilled water of 2 times of parts by volume and dialyse, merge dialysis solution, vacuum concentration; Dialysis solution added in the DEAE-23 cellulose chromatography post crosses post and separate, use the distilled water eluting, collect eluent up to ultraviolet absorption peak till 262nm place does not have absorption, concentrate eluant; Concentrated solution is separated through the Dowex50WX8 resin column, use 0.25MNH
4The OH eluting, collect eluent up to ultraviolet absorption peak at 262nm place not till the absorption, this eluent is concentrated into thick (remaining 20 parts by volume), put into the lyophilization of vacuum lyophilization instrument, condition is-40 ℃ to-35 ℃, 12-15Pa, 28-32h, obtains the sticking slightly material GTF of yellowish-brown activity extract.
The preparation of embodiment 8:GTF activity extract
Take by weighing 30g DEAE-23 celluosic resin (DEAE-cellulose) dry powder, join in the 0.2NNaOH solution of about 160ml, the limit edged stirs, place and soak after 0.6 hour in the beaker, use the distilled water eluting, till washing neutrality, join again in the 0.2NHCL solution of 160 parts by volume, use with quadrat method and wash neutrality; The processing of getting Dowex50WX8 resin (732 strong acidic ion resin) is with the DEAE-23 celluosic resin; Take by weighing the Rich chromium yeast powder of 20g, the dose volume ratio is 1: 1 n-butanol-water mixture 400ml, and yeast powder is added in the above-mentioned mixed liquor, stirs 3 hours, leaves standstill 20h, and the supernatant of getting aqueous phase is at 60 ℃ of following vacuum concentration; Concentrated solution is added respectively in the distilled water of 2 times of parts by volume and dialyse, merge dialysis solution, vacuum concentration; Dialysis solution added in the DEAE-23 cellulose chromatography post crosses post and separate, use the distilled water eluting, collect eluent up to ultraviolet absorption peak till 262nm place does not have absorption, concentrate eluant; Concentrated solution is separated through the Dowex50WX8 resin column, use 0.25MNH
4The OH eluting, collect eluent up to ultraviolet absorption peak at 262nm place not till the absorption, this eluent is concentrated into thick (remaining 10 parts by volume), put into the lyophilization of vacuum lyophilization instrument, condition is-40 ℃ to-35 ℃, 12-15Pa, 28-32h, obtains the sticking slightly material GTF of yellowish-brown activity extract.
The preparation of embodiment 9:GTF activity extract
The preparation technology of GTF activity extract is: take by weighing 30g DEAE-23 celluosic resin (DEAE-cellulose) dry powder, join in the 0.2NNaOH solution of about 140ml, the limit edged stirs, place and soak after 0.5 hour in the beaker, use the distilled water eluting, till washing neutrality, join again in the 0.2NHCL solution of 140 parts by volume, use with quadrat method and wash neutrality; The processing of getting Dowex50WX8 resin (732 strong acidic ion resin) is with DEAE cellulose-23; Take by weighing the Rich chromium yeast powder of 40g, the dose volume ratio is 1: 1 n-butanol-water mixture 800ml, and yeast powder is added in the above-mentioned mixed liquor, stirs 4 hours, leaves standstill 20h, and the supernatant of getting aqueous phase is at 50 ℃ of following vacuum concentration; Concentrated solution is added respectively in the distilled water of 2 times of parts by volume and dialyse, merge dialysis solution, vacuum concentration; Dialysis solution added in the DEAE-23 cellulose chromatography post crosses post and separate, use the distilled water eluting, collect eluent up to ultraviolet absorption peak till 262nm place does not have absorption, concentrate eluant; Concentrated solution is separated through the Dowex50WX8 resin column, use 0.25MNH
4The OOH eluting, collect eluent up to ultraviolet absorption peak at 262nm place not till the absorption, this eluent is concentrated into thick (remaining 30 parts by volume), put into the lyophilization of vacuum lyophilization instrument, condition is-40 ℃ to-35 ℃, 12-15Pa, 28-32h, obtains the sticking slightly material GTF of yellowish-brown activity extract.
Embodiment 10: the preparation of capsule
Get GTF activity extract 47g and mix with chitin 33g, dextrin 20g respectively, promptly obtain the dried medicated powder that 100g contains GTF activity extract active component, the medicated powder that makes is made every 0.2g of capsule according to conventional method, once a day, each one.
Embodiment 11: the preparation of tablet
Get GTF activity extract 17g and mix with chitin 53g, dextrin 30g respectively, promptly obtain the dried medicated powder that 100g contains GTF activity extract active component, the medicated powder that makes is made tablet according to conventional method.Every 0.5g, once a day, each a slice.
Embodiment 12: the preparation of oral liquid
Get GTF activity extract 0.9g, be dissolved in the 91ml pure water, make oral liquid according to conventional method, every 10ml, once a day, each one.
Embodiment 13: the preparation of drop pill
Get GTF activity extract 47g and mix with chitin 33g, dextrin 20g respectively, promptly obtain the dried medicated powder that 100g contains GTF activity extract active component, the medicated powder that makes is made drop pill according to conventional method.
Claims (21)
1. a pharmaceutical composition that contains the GTF activity extract is characterized in that the raw material of this pharmaceutical composition consists of: GTF activity extract 10-50 weight portion, chitin 30-60 weight portion, dextrin 20-35 weight portion.
2. pharmaceutical composition as claimed in claim 1 is characterized in that the raw material of this pharmaceutical composition consists of: GTF activity extract 30-47 weight portion, chitin 33-43 weight portion, dextrin: 20-27 weight portion.
3. pharmaceutical composition as claimed in claim 1 is characterized in that the raw material of this pharmaceutical composition consists of: GTF activity extract 24-34 weight portion, chitin 41-47 weight portion, dextrin 25-29 weight portion.
4. pharmaceutical composition as claimed in claim 1 is characterized in that the raw material of this pharmaceutical composition consists of: GTF activity extract 12-17 weight portion, chitin 53-55 weight portion, dextrin 30-33 weight portion.
5. pharmaceutical composition as claimed in claim 1 is characterized in that the raw material of this pharmaceutical composition consists of: GTF activity extract 47 weight portions, chitin 33 weight portions, dextrin 20 weight portions.
6. pharmaceutical composition as claimed in claim 1 is characterized in that the raw material of this pharmaceutical composition consists of: GTF activity extract 35 weight portions, chitin 40 weight portions, dextrin 25 weight portions.
7. pharmaceutical composition as claimed in claim 1 is characterized in that the raw material of this pharmaceutical composition consists of: GTF activity extract 17 weight portions, chitin 53 weight portions, dextrin 30 weight portions.
8. as the arbitrary described pharmaceutical composition of claim 1-7, it is characterized in that said composition, make tablet, granule, capsule, oral liquid, drop pill or the injection of clinical acceptance according to common process.
9. as the arbitrary described pharmaceutical composition of claim 1-7, it is characterized in that the GTF activity extract is made by following method in this pharmaceutical composition:
Take by weighing 20-40 weight portion weak-base ion-exchange resin dry powder, join earlier in the 0.2-1 equivalent NaOH solution of 130-170 parts by volume, the limit edged stirs, place and soak after 0.5-1.0 hour in the beaker, use the distilled water eluting, be washed till neutrality, join again in the 0.2-2 equivalent HCL solution of 130-170 parts by volume, be washed till neutrality with quadrat method; Get strong acidic ion resin dry powder 20-40 weight portion, processing method is with weak-base ion-exchange resin dry powder;
Take by weighing the Rich chromium yeast powder of 20-40 weight portion, dose volume adds yeast powder in the above-mentioned mixed liquor than being the n-butanol-water mixed liquor 400-800 parts by volume of 0.5-1.5: 0.5-1.5, stirs 2-4 hour, left standstill 12-20 hour, the supernatant of getting aqueous phase is at 50 ℃ of-60 ℃ of following vacuum concentration; The distilled water that concentrated solution is added 2-4 times of parts by volume is dialysed, and merges dialysis solution, vacuum concentration; Dialysis solution after will concentrating adds to be crossed post and separates in the weak-base ion-exchange resin chromatographic column, and use the distilled water eluting, collect eluent up to ultraviolet absorption peak till 262nm place does not have absorption, concentrate eluant; Concentrated solution is separated through the strong acidic ion resin post, with 0.25 mole of NH
4The OH eluting, till the 262nm place did not absorb, it was the 10-30 parts by volume that this eluent is concentrated into residual volume to the collection eluent up to ultraviolet absorption peak, lyophilization obtains the GTF activity extract.
10. pharmaceutical composition as claimed in claim 9 is characterized in that wherein cryodesiccated freezing conditions is that temperature is that-40 ℃ to-35 ℃, pressure are that 12-15Pa, time are 28-32h.
11. pharmaceutical composition as claimed in claim 9 is characterized in that weak-base ion-exchange resin wherein is meant the celluosic resin of DEAE-23, DEAE-11, DEAE-22, DEAE-23, DEAE-32 or DEAE51-53 model; Strong acidic ion resin is meant the resin of Dowex50W-X8 Dowex50W-X2, X4, X8, X12 model.
12. as claim 10 or 11 described pharmaceutical compositions, it is characterized in that the GTF activity extract is made by following method in this pharmaceutical composition: take by weighing 30 weight portion DEAE-23 celluosic resin dry powder, join earlier in the 0.2 equivalent NaOH solution of about 150 parts by volume, the limit edged stirs, place in the beaker and soak after 0.6 hour, use the distilled water eluting, till washing neutrality, join again in the 0.2 equivalent HCL solution of 150 parts by volume, be washed till neutrality with quadrat method; Get the Dowex50W-X8 resin, its processing method is with the DEAE-23 celluosic resin; Take by weighing the Rich chromium yeast powder of 30 weight portions, the dose volume ratio is 1: 1 n-butanol-water mixture 600 parts by volume, and yeast powder is added in the above-mentioned mixed liquor, stirs 3 hours, leaves standstill 20h, and the supernatant of getting aqueous phase is at 55 ℃ of following vacuum concentration; The distilled water that concentrated solution is added 2 times of parts by volume is respectively dialysed, and merges dialysis solution, vacuum concentration; Dialysis solution added in the DEAE-23 celluosic resin chromatography post crosses post and separate, use the distilled water eluting, collect eluent up to ultraviolet absorption peak till 262nm place does not have absorption, concentrate eluant; Concentrated solution is separated through the Dowex50WX8 cationic resin column, with 0.25 mole of NH
4The OH eluting, the collection eluent till the 262nm place does not absorb, is concentrated into residue 20 parts by volume with this eluent up to ultraviolet absorption peak, puts into the lyophilization of vacuum lyophilization instrument, condition is-40 ℃ to-35 ℃, 12-15Pa, 28-32h, obtains the GTF activity extract of yellowish-brown viscosity.
13., it is characterized in that Huo Chrome content is not less than 1360mg/Kg in the wherein said GTF activity extract as claim 1,2,3,4,5,6,7,10 or 11 described pharmaceutical compositions.
14. the application in the medicine of, diabetes low at preparation treatment carbohydrate tolerance or hyperlipemia as claim 1,2,3,4,5,6,7,10 or 11 described pharmaceutical compositions.
15. pharmaceutical composition as claimed in claim 12 is low at preparation treatment carbohydrate tolerance, the application in the medicine of diabetes or hyperlipemia.
16. the preparation method of a GTF activity extract is characterized in that this method is:
Take by weighing 20-40 weight portion weak-base ion-exchange resin dry powder, join earlier in the 0.2-1 equivalent NaOH solution of 130-170 parts by volume, the limit edged stirs, place and soak after 0.5-1.0 hour in the beaker, use the distilled water eluting, be washed till neutrality, join again in the 0.2-2 equivalent HCL solution of 130-170 parts by volume, be washed till neutrality with quadrat method; Get strong acidic ion resin dry powder 20-40 weight portion, processing method is with weak-base ion-exchange resin dry powder;
Take by weighing the Rich chromium yeast powder of 20-40 weight portion, dose volume adds yeast powder in the above-mentioned mixed liquor than being the n-butanol-water mixed liquor 400-800 parts by volume of 0.5-1.5: 0.5-1.5, stirs 2-4 hour, left standstill 12-20 hour, the supernatant of getting aqueous phase is at 50 ℃ of-60 ℃ of following vacuum concentration; The distilled water that concentrated solution is added 2-4 times of parts by volume is dialysed, and merges dialysis solution, vacuum concentration; Dialysis solution after will concentrating adds to be crossed post and separates in the weak-base ion-exchange resin chromatographic column, and use the distilled water eluting, collect eluent up to ultraviolet absorption peak till 262nm place does not have absorption, concentrate eluant; Concentrated solution is separated through the strong acidic ion resin post, with 0.25 mole of NH
4The OH eluting, till the 262nm place did not absorb, it was the 10-30 parts by volume that this eluent is concentrated into residual volume to the collection eluent, lyophilization up to ultraviolet absorption peak, obtain the GTF activity extract, active road content is not less than 1360mg/Kg in the GTF activity extract.
17. the preparation method of GTF activity extract as claimed in claim 16 is characterized in that this method is:
Take by weighing 20-40 weight portion weak-base ion-exchange resin dry powder, join earlier in the 0.2-1 equivalent NaOH solution of 130-170 parts by volume, the limit edged stirs, place and soak after 0.5-1.0 hour in the beaker, use the distilled water eluting, be washed till neutrality, join again in the 0.2-2 equivalent HCL solution of 130-170 parts by volume, be washed till neutrality with quadrat method; Get strong acidic ion resin dry powder 20-40 weight portion, processing method is with weak-base ion-exchange resin dry powder;
Take by weighing the Rich chromium yeast powder of 20-40 weight portion, dose volume adds yeast powder in the above-mentioned mixed liquor than being the n-butanol-water mixed liquor 400-800 parts by volume of 0.5-1.5: 0.5-1.5, stirs 2-4 hour, left standstill 12-20 hour, the supernatant of getting aqueous phase is at 50 ℃ of-60 ℃ of following vacuum concentration; The distilled water that concentrated solution is added 2-4 times of parts by volume is dialysed, and merges dialysis solution, vacuum concentration; Dialysis solution after will concentrating adds to be crossed post and separates in the weak-base ion-exchange resin chromatographic column, and use the distilled water eluting, collect eluent up to ultraviolet absorption peak till 262nm place does not have absorption, concentrate eluant; Concentrated solution is separated through the strong acidic ion resin post, with 0.25 mole of NH
4The OH eluting, till the 262nm place did not absorb, it was the 10-30 parts by volume that this eluent is concentrated into residual volume to the collection eluent, lyophilization up to ultraviolet absorption peak, obtain the GTF activity extract, Huo Chrome content is not less than 1360mg/Kg in the GTF activity extract.
18. pharmaceutical composition as claimed in claim 9, it is characterized in that the GTF activity extract is made by following method in this pharmaceutical composition: take by weighing 30 weight portion DEAE-23 celluosic resin dry powder, join earlier in the 0.2 equivalent NaOH solution of about 150 parts by volume, the limit edged stirs, place in the beaker and soak after 0.6 hour, use the distilled water eluting, till washing neutrality, join again in the 0.2 equivalent HCL solution of 150 parts by volume, be washed till neutrality with quadrat method; Get the Dowex50W-X8 resin, its processing method is with the DEAE-23 celluosic resin; Take by weighing the Rich chromium yeast powder of 30 weight portions, the dose volume ratio is 1: 1 n-butanol-water mixture 600 parts by volume, and yeast powder is added in the above-mentioned mixed liquor, stirs 3 hours, leaves standstill 20h, and the supernatant of getting aqueous phase is at 55 ℃ of following vacuum concentration; The distilled water that concentrated solution is added 2 times of parts by volume is respectively dialysed, and merges dialysis solution, vacuum concentration; Dialysis solution added in the DEAE-23 celluosic resin chromatography post crosses post and separate, use the distilled water eluting, collect eluent up to ultraviolet absorption peak till 262nm place does not have absorption, concentrate eluant; Concentrated solution is separated through the Dowex50WX8 cationic resin column, with 0.25 mole of NH
4The OH eluting, the collection eluent till the 262nm place does not absorb, is concentrated into residue 20 parts by volume with this eluent up to ultraviolet absorption peak, puts into the lyophilization of vacuum lyophilization instrument, condition is-40 ℃ to-35 ℃, 12-15Pa, 28-32h, obtains the GTF activity extract of yellowish-brown viscosity.
19. pharmaceutical composition as claimed in claim 9 is characterized in that Huo Chrome content is not less than 1360mg/Kg in the wherein said GTF activity extract.
20. pharmaceutical composition as claimed in claim 9 is low at preparation treatment carbohydrate tolerance, the application in the medicine of diabetes or hyperlipemia.
21. pharmaceutical composition as claimed in claim 9 is low at preparation treatment carbohydrate tolerance, the application in the medicine of diabetes or hyperlipemia.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2006101042774A CN101120954A (en) | 2006-08-09 | 2006-08-09 | Medicinal composition containing GTF active extract and its preparation technology |
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| CNA2006101042774A CN101120954A (en) | 2006-08-09 | 2006-08-09 | Medicinal composition containing GTF active extract and its preparation technology |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102573782A (en) * | 2009-03-27 | 2012-07-11 | 赫尔克里士公司 | Aminated polymers and their use in water-borne compositions |
| CN101637145B (en) * | 2009-08-27 | 2012-08-22 | 山西中医学院 | Biotransformation high GTF activity milk and manufacturing method thereof |
-
2006
- 2006-08-09 CN CNA2006101042774A patent/CN101120954A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102573782A (en) * | 2009-03-27 | 2012-07-11 | 赫尔克里士公司 | Aminated polymers and their use in water-borne compositions |
| CN101637145B (en) * | 2009-08-27 | 2012-08-22 | 山西中医学院 | Biotransformation high GTF activity milk and manufacturing method thereof |
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