CN108186691A - Application of the okamura ishige algae extract in alpha-glucosidase restrainer is prepared - Google Patents
Application of the okamura ishige algae extract in alpha-glucosidase restrainer is prepared Download PDFInfo
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- CN108186691A CN108186691A CN201810162191.XA CN201810162191A CN108186691A CN 108186691 A CN108186691 A CN 108186691A CN 201810162191 A CN201810162191 A CN 201810162191A CN 108186691 A CN108186691 A CN 108186691A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/02—Algae
- A61K36/03—Phaeophycota or phaeophyta (brown algae), e.g. Fucus
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
- A61K9/1605—Excipients; Inactive ingredients
- A61K9/1629—Organic macromolecular compounds
- A61K9/1652—Polysaccharides, e.g. alginate, cellulose derivatives; Cyclodextrin
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/15—Preparation or pretreatment of starting material involving mechanical treatment, e.g. chopping up, cutting or grinding
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
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Abstract
The present invention discloses a kind of application of okamura ishige algae extract in α glucosidase inhibitors are prepared, by okamura ishige algae water extract-alcohol precipitation supernatant fraction and soluble starch, honey element and beta-cyclodextrin carry out the preparation of okamura ishige algae electuary in proportion, and the blood sugar reducing function of okamura ishige algae electuary is evaluated, research shows that, okamura ishige algae extract has α glucosidase activities certain inhibiting effect, and exploitation okamura ishige algae extract has feasibility as α glucosidase inhibitors.
Description
Technical field
The present invention relates to pharmaceutical preparations technology fields, and in particular to be that a kind of okamura ishige algae extract is preparing phlorose
Application in glycosides enzyme inhibitor.
Background technology
Diabetes are a kind of common metabolic disease groups characterized by chronic blood glucose level increases, and long-term blood glucose increases,
Easily make vascular injury and jeopardize the organs such as the heart, brain, kidney, eye, seriously affect human life safety.For this purpose, people are dedicated to studying
Inhibit the drug of blood glucose rise.At present, alpha-glucosidase restrainer such as acarbose etc. inhibits starch to decompose due to having
For the rate of glucose, slow down enteron aisle to effectiveness such as the absorptions of carbohydrate, it is considered to be preferred oral hypoglycemic drug.But
Clinical application is likely to result in the side effects such as flatulence, diarrhea, abdominal pain it has also been found that the new orals antidiabetic drug such as acarbose tablet.Cause
This, safer efficient novel alpha-glucosidase restrainer is found from edible, medicinal plant and is constantly subjected to learn extensively
The favor of person.
It is peaceful to be distributed in brown alga okamura ishige algae (Ishige okamurae) system north of southeastern coast middle and high tide in China's with rock reef
The distinctive edible seaweed in madder portion, methanolic extract have the polyphenol content of high level, the petroleum ether of methanolic extract, two
Five extraction parts such as chloromethanes, ethyl acetate, n-butanol and water are respectively provided with certain antioxidant activity and alpha-glucosidase
Inhibitory activity, 80% methanolic extract can by change hepatic glucose metabolic enzyme activity, improve insulin resistance play it is hypoglycemic
Effect, have important nutritive value and potential healthcare function.And okamura ishige algae extract presses down alpha-glucosidase activity
The correlative study of making is not yet reported.
Invention content
The purpose of the present invention is to provide a kind of application of okamura ishige algae extract in alpha-glucosidase restrainer is prepared.
In order to achieve the above objectives, solution of the invention is：
Application of the okamura ishige algae extract in alpha-glucosidase restrainer is prepared.
The okamura ishige algae extract is prepared by following method：
(1) it first removes after miscellaneous fresh okamura ishige algae cleans and drain, okamura ishige algae powder is ground into through tissue pulverizer；
(2) and then the okamura ishige algae powder after crushing is taken, according to okamura ishige algae powder:Distilled water=1g:The ratio mixing of 30mL,
Twice, 1~2h, suction filtration take filtrate every time for water-bath extraction at 90~100 DEG C；
(3) and then by gained filtrate decompression the 23%~43% of filtrate original volume is concentrated into, adding in absolute ethyl alcohol makes mixing
In system the volume fraction of absolute ethyl alcohol be 75%, at 4 DEG C alcohol precipitation for 24 hours after, centrifugation, obtain supernatant and alcohol hypostasis；
(4) after supernatant is taken to be concentrated under reduced pressure, through vacuum freeze drying, okamura ishige algae water extract-alcohol precipitation supernatant fraction, i.e. this hair are obtained
The bright okamura ishige algae extract.
The reduced pressure carries out under conditions of 65 DEG C, -0.09MPa.
The rotating speed of the centrifugation is 4000r/min, and the time of the centrifugation is 5min.
The temperature of the vacuum freeze drying is -80 DEG C.
The dosage form of the okamura ishige algae extract is okamura ishige algae electuary, and the preparation process of the okamura ishige algae electuary is：By described in
Okamura ishige algae extract, soluble starch, honey element, cycloheptaamylose are according to 20~40:50~70:0.5~2:6.5~8.5
After quality proportioning mixing, add appropriate 95% ethyl alcohol, be made pinch can the agglomerating, softwood that can scatter of pressure, cross 40 mesh medicine sieve series grains, granulation
By 80 DEG C of dryings, whole grain, ultraviolet sterilization, pack to get the okamura ishige algae electuary.
After adopting the above technical scheme, okamura ishige algae extract of the present invention answering in alpha-glucosidase restrainer is prepared
With, research shows that, okamura ishige algae extract has certain inhibiting effect to alpha-glucosidase activity, develops okamura ishige algae extract
There is feasibility as alpha-glucosidase restrainer.
Description of the drawings
Fig. 1 is the result schematic diagram that okamura ishige algae electuary inhibits alpha-glucosidase activity.
Specific embodiment
In order to further explain the technical solution of the present invention, the present invention is explained in detail below by specific embodiment
It states.
Embodiment
1st, the preparation of okamura ishige algae extract
(1) it first removes after miscellaneous fresh okamura ishige algae cleans and drain, okamura ishige algae powder is ground into through tissue pulverizer；
(2) and then the okamura ishige algae powder after crushing is taken, according to okamura ishige algae powder:Distilled water=1g:The ratio mixing of 30mL,
Twice, 1~2h, suction filtration take filtrate every time for water-bath extraction at 90~100 DEG C；
(3) and then under 65 DEG C, -0.09MPa, gained filtrate decompression is concentrated into the 33% of filtrate original volume, adds in nothing
Water-ethanol makes the volume fraction of absolute ethyl alcohol in mixed system be 75%, at 4 DEG C alcohol precipitation for 24 hours after, then with 4000r/min's
Rotating speed centrifuges 5min, obtains supernatant and alcohol hypostasis；
(4) under 65 DEG C, -0.09MPa, after supernatant is taken to be concentrated under reduced pressure, through -80 DEG C of vacuum freeze dryings, okamura ishige algae is obtained
Water extract-alcohol precipitation supernatant fraction, i.e. okamura ishige algae extract of the invention.
2nd, the preparation of okamura ishige algae electuary
By obtained okamura ishige algae extract, soluble starch, honey element, cycloheptaamylose according to 30:60:1:7.5 matter
After amount proportioning mixing, add appropriate 95% ethyl alcohol, be made pinch can the agglomerating, softwood that can scatter of pressure, 40 mesh medicine sieve series grains are crossed, after granulation
Through 80 DEG C of dryings, whole grain, ultraviolet sterilization, pack to get okamura ishige algae electuary.
3rd, the measure that okamura ishige algae electuary inhibits alpha-glucosidase activity
3.1 assay method
With reference to [LI T, ZHANG XD, SONG YW, et al.A microplate-based screening such as Li T
method for alpha-glucosidase inhibitors[J].Chin.J.Clin.Pharm.Ther.,2005,10
(10):1129-1134.] method, 112 μ L 0.1mol/L PBS (pH=6.8), 20 μ L are sequentially added in 96 hole elisa Plates
0.2U/mL alpha-glucosidases, 8 μ L sample liquids (are prepared) with dimethyl sulfoxide (DMSO), and 2.5mmol/ is added in after 37 DEG C of constant temperature 15min
The Na of 80 μ L 0.2mol/L is added in after L 4- nitrobenzene-α-D- glucopyranosides (PNPG) 20 μ L, reacted 15min2CO3It is molten
Liquid terminates reaction, and absorbance value is surveyed under microplate reader 405nm wavelength, repeats experiment 3 times, is averaged.The public affairs of inhibition of enzyme activity rate
Formula：I%=[1- (A1- A2)/(A3- A4)] × 100 ﹪；In formula：A1For the absorbance value of sample liquid group, A2To replace enzyme with PBS
The sample liquid group Background absorbance value that solution measures, A3For the blank control group absorbance value measured with DMSO instead of sample liquid, A4For with
DMSO replaces the blank control group Background absorbance value that sample liquid replaces enzyme solutions to measure with PBS.
3.2 results and analysis
Okamura ishige algae electuary is to influence result such as Fig. 1 of alpha-glucosidase figure activity.As shown in Figure 1, okamura ishige algae electuary pair
Alpha-glucosidase activity shows as stronger inhibiting effect, and during sample liquid concentration 0.50mg/mL, enzyme activity inhibiting rate is reachable
87.95%, in the testing concentration range of 0.17~0.50mg/mL, dose-effect meets equation of linear regression y=269.04x-
50.185(R2=0.977, P<0.01) extremely significant positive correlation (P, is presented<0.01), 503nhibiting concentration IC50For 0.37mg/mL,
Show that the okamura ishige algae electuary has stronger alpha-glucosaccharase enzyme inhibition activity.
4th, okamura ishige algae electuary causes alloxan the blood sugar reducing function of diabetic mouse model
4.1 test method
Male, 22 ± 1g of weight SPF grades health KM mouse (Shanghai Slac Experimental Animal Co., Ltd. offer,
Credit number is SCXK (Shanghai) 2012-0002) it is fed 3 days in mouse room adaptability, appetite is appropriate, and drinking-water is sufficient, keeps mouse room temperature
18~20 DEG C of degree.12h is deprived of food but not water after 3 days, 2% alloxan solution (sterile physiological is injected intraperitoneally by 200mg/kg dosage
Saline), it randomly selects 10 mouse and isometric physiological saline is injected intraperitoneally as normal group (NC).Restore after 1h normal
Diet, after 3d docking take blood (mouse is deprived of food but not water 12h before taking blood), blood glucose meter (the stable YZBI Hunan 004-2012 of three promises, Changsha
Three promise biosensor technique limited companies) blood glucose value is surveyed, it surveys a blood glucose again every other day, with blood glucose value stabilization and is more than
11.1mmol/L and the more drinks of appearance, mostly food, the mouse of diuresis phenomenon, which are considered as, to be modeled successfully.
Successful diabetic mice will be modeled and be divided into 5 groups by weight, blood glucose, each group animal blood glucose mean difference is not more than
0.5mmol/L, every group 10, be denoted as respectively diabetes model control group (MC), positive control Metformin hydrochloride group (MH,
500mg/kg), okamura ishige algae electuary low dose group (LI, 350mg/kg), okamura ishige algae electuary middle dose group (MI, 700mg/kg), iron
Follow closely dish electuary high dose group (HI, 1400mg/kg).According to dosage gastric infusion is primary during administration group every afternoon 5, normal group and mould
The physiological saline of equivalent, continuous 14d is given once daily in type control group.A fasting blood-glucose is surveyed per 7d (to be deprived of food but not water after dinner
12h measures blood glucose value between 7~8 o'clock of the next morning).Experiment end, surveys according to dosage gavage 1 time after fasting blood sugar, 20min
Gavage glucose 2.0g/kg afterwards is measured to the blood glucose value of 0.5h, 1h, 2h after glucose.
4.2 result and analysis
The blood sugar reducing function of diabetic mouse model research okamura ishige algae electuary, experimental result such as table 1 are caused with alloxan.By table
1 it is found that compared with model group, and nail nail each dosage group of dish electuary has certain hypoglycemic effect in observation time-histories 7d and 14d,
Difference has extremely significant statistical significance (P<, and the hypoglycemic of each dosage group blood sugar reducing function and Metformin hydrochloride group 0.01)
Difference between the effects do not have statistical significance (P>0.05), blood sugar reducing function difference does not have statistics meaning yet between each dosage group during 14d
Justice (P>0.05), illustrate that okamura ishige algae electuary can effectively play blood sugar decreasing effect in the dosage of 350mg/kg.
1 okamura ishige algae electuary of table causes alloxan the influence (n=10 of diabetic mouse model blood glucose value；##P<0.01)
In general, after oral glucose, serum level of glucose raising peaking during 0.5~1h, restore after 2h to
Fasting blood sugar.As seen from Table 2, in each dosage group of okamura ishige algae electuary mouse after oral glucose, each time point
Blood glucose value is substantially less than the blood glucose value (P of model control group (MC)<0.01), illustrate that okamura ishige algae electuary can drop in a short time
Low postprandial blood sugar, and its hypoglycemic effect (P close with positive drug melbine>0.05).
To sum up research shows that：Okamura ishige algae extract has certain inhibiting effect to alpha-glucosidase activity, develops iron
Follow closely dish extract has feasibility as alpha-glucosidase restrainer.
2 okamura ishige algae electuary of table causes alloxan the influence (n=10 of diabetic mouse model sugar tolerance；##P<0.01)
The product form and style of above-described embodiment and schema and the non-limiting present invention, any technical field it is common
The appropriate variation or modification that technical staff does it all should be regarded as not departing from the patent category of the present invention.
Claims (6)
1. application of the okamura ishige algae extract in alpha-glucosidase restrainer is prepared.
2. application of the okamura ishige algae extract according to claim 1 in alpha-glucosidase restrainer is prepared, feature
It is：The okamura ishige algae extract is prepared by following method：
(1) it first removes after miscellaneous fresh okamura ishige algae cleans and drain, okamura ishige algae powder is ground into through tissue pulverizer；
(2) and then the okamura ishige algae powder after crushing is taken, according to okamura ishige algae powder:Distilled water=1g:The ratio mixing of 30mL, 90~
Twice, 1~2h, suction filtration take filtrate every time for water-bath extraction at 100 DEG C；
(3) and then by gained filtrate decompression the 23%~43% of filtrate original volume is concentrated into, adding in absolute ethyl alcohol makes mixed system
The volume fraction of middle absolute ethyl alcohol be 75%, at 4 DEG C alcohol precipitation for 24 hours after, centrifugation, obtain supernatant and alcohol hypostasis；
(4) after supernatant is taken to be concentrated under reduced pressure, through vacuum freeze drying, okamura ishige algae water extract-alcohol precipitation supernatant fraction, i.e., described iron nail are obtained
Dish extract.
3. application of the okamura ishige algae extract according to claim 2 in alpha-glucosidase restrainer is prepared, feature
It is：The reduced pressure carries out under conditions of 65 DEG C, -0.09MPa.
4. application of the okamura ishige algae extract according to claim 2 in alpha-glucosidase restrainer is prepared, feature
It is：In step (3), the rotating speed of the centrifugation is 4000r/min, and the time of the centrifugation is 5min.
5. application of the okamura ishige algae extract according to claim 2 in alpha-glucosidase restrainer is prepared, feature
It is：In step (4), the temperature of the vacuum freeze drying is -80 DEG C.
6. application of the okamura ishige algae extract according to claim 1 in alpha-glucosidase restrainer is prepared, feature
It is：The dosage form of the okamura ishige algae extract is okamura ishige algae electuary, and the preparation process of the okamura ishige algae electuary is by the iron nail
Dish extract is with soluble starch, honey element, cycloheptaamylose according to 20~40:50~70:0.5~2:6.5~8.5 quality
After proportioning mixing, add appropriate 95% ethyl alcohol, be made softwood, cross 40 mesh medicine sieve series grains, through 80 DEG C of dryings, whole grain, ultraviolet sterilization, close
Package is filled to get the okamura ishige algae electuary.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20100039104A (en) * | 2008-10-07 | 2010-04-15 | 부경대학교 산학협력단 | An anti-oxidant active composition containing compounds from ishige okamurae |
| CN106892999A (en) * | 2017-04-24 | 2017-06-27 | 浙江海洋大学 | A kind of brown alga okamura ishige algae extraction method of polysaccharides |
-
2018
- 2018-02-27 CN CN201810162191.XA patent/CN108186691A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20100039104A (en) * | 2008-10-07 | 2010-04-15 | 부경대학교 산학협력단 | An anti-oxidant active composition containing compounds from ishige okamurae |
| CN106892999A (en) * | 2017-04-24 | 2017-06-27 | 浙江海洋大学 | A kind of brown alga okamura ishige algae extraction method of polysaccharides |
Non-Patent Citations (1)
| Title |
|---|
| 黄晓冬等: "海藻铁钉菜抑制α-葡萄糖苷酶活性部位的筛选及其反应动力学研究", 《中国医院药学杂志》 * |
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