CN100998865A - Modicine and food dual-purpose composition using forest frog active peptide component as active constituent and its use - Google Patents
Modicine and food dual-purpose composition using forest frog active peptide component as active constituent and its use Download PDFInfo
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Abstract
A medical composition containing the active peptide of forest frog as its active component and used a both medicine and food for preventing and treating constipation, anxiety, hyperlipemia, hyperglycemia and climacteric syndrome, delaying sanility, beautifying skin and relieving fatigue is prepared from said active peptide of forest frog and the pharmacologically and bromatologically acceptable auxiliary.
Description
Technical field
The present invention relates to wood frog bioactive peptide component is dietotherapeutic preparation method of composition of active component and uses thereof, belongs to medicine and health food technology field.
Background technology
Along with society and expanding economy, variation has taken place in people's work and life style, cause thereupon such as senilism, climacteric in advance, incidence rates such as gastrointestinal dysfunction, fatigue syndrome, immunity degradation, anxiety, blood glucose and dyslipidemia increase.Modern Chinese medicine and biomedical research find that some natural bioactive ingredients plays key effect aspect preventing and curing diseases.The wood frog bioactive peptide is to be organized through chromatography purification or the synthetic peptide matters of application modern molecular biology technique by Oviductus Ranae, Rana sylvatica Le conte skin etc., finds that recently it has biological effect widely.Wood frog has multiple medicinal and dietetic therapy effect, the dietotherapeutic tonic of Chang Zuowei strengthening by means of tonics among the people, modern pharmacological research confirms that some extract of wood frog has the cell of enhancing and humoral immunization, blood fat reducing and blood glucose, relieving cough and expelling phlegm, defying age, fatigue-resisting function, and experiment shows that the peptide matters that is obtained through conventional chromatography purification by tissues such as Oviductus Ranae, Rana sylvatica Le conte skins has slow down aging, improves looks, delays and alleviate the function of climacteric syndrome, prophylactic treatment constipation, resisting fatigue, raising immunity, anxiety, blood biochemical adjusting.Therefore, be active component with wood frog bioactive peptide component, mix the dietotherapeutic compositions that forms with acceptable auxiliary on pharmacy and the Food technology, pharmacology and health-care effect are clear and definite, and preparation technology is simple, is easy to promote.
Publication number is the Chinese patent of CN1216768, discloses the preparation method of wood frog active antibacterial peptide, antibacterial peptide is not prepared into medicine or health food; Publication number is the Chinese patent of CN153165A, CN153166A, discloses the gel and the spray of wood frog active antibacterial peptide respectively, only is used for clinical scald, skin infection external and the sterilization of daily household.
Summary of the invention
We studies confirm that the wood frog bioactive peptide at slow down aging, improve looks, delay to alleviate the potential of aspects such as climacteric syndrome, prophylactic treatment constipation, resisting fatigue, raising immunity, anxiety, blood glucose and blood lipid regulation.
Promote the experiment of skin epithelial cell proliferation: the rat skin epithelial cell of cultivating to go down to posterity is a cell model research wood frog bioactive peptide to the active influence of rising in value of skin epithelial cell.Result of study is found to cultivate the rat skin epithelial cell 12-48 hour with containing 0.01% through the culture fluid through the wood frog bioactive peptide component of screening of aseptic sucking filtration, experimental group cultured cell form is significantly better than matched group, mtt assay test experiments group cells survival activity is apparently higher than matched group and have statistical significant difference (p＜0.05), sees Table 1.
The rat skin epithelial cell active influence (mtt assay) of table 1. wood frog bioactive peptide to going down to posterity and cultivating
| Incubation time | The OD value | |
| Matched group (n=8) | Experimental group (n=8) | |
| 12 hours 24 hours 36 hours 48 hours | 0.446±0.012 0.450±0.023 0.489±0.031 0.437±0.024 | 0.439±0.005 0.525±0.041 * 0.561±0.034 * 0.532±0.087 * |
Annotate: * and matched group compare, p＜0.05
Defying age, anxiety and resisting fatigue experiment: 40 of kunming mices, 22-26g, basic forage feed is equally divided into 4 groups, and each is organized and gives 0.00,0.25,0.50 respectively every day, the wood frog bioactive peptide component of 1.0g/kg.bw is irritated 6 weeks of stomach, carries out following experiment:
Mice diving tower experiment (strengthening the ability of learning and memory experiment) diving tower reflective box bottom is 30V continuous voltage electromagnetic pole copper grid, built-in height of reflective box and diameter are the platform of 4.5cr, animal is put into reflective box endoadaptation 3min, energising immediately then, after being shocked by electricity, animal jumps onto platform, most animals may be once more or is repeatedly skipped on the copper grid, is subjected to the platform that snaps back again that shocks by electricity.Training 3min, the number of times that the record mice is shocked by electricity is done memory and is kept test behind the 24h, and the incubation period that record jumps off platform for the first time, the number of times that is shocked by electricity in record incubation period and the 5min is tested learning and memory abilities in aging mice with this.Result's (seeing Table 2) shows that the wood frog bioactive peptide can strengthen learning and memory abilities in aging mice.
Table 2. wood frog bioactive peptide component is to the influence of ability of learning and memory in mice
N=10)
| (g/kg·bw) | Incubation period (s) | Errors number |
| 0.00 0.25 0.50 1.00 | 289.1±55.4 312.8±83.2 322.3±71.5 356.2±95.2 * | 2.1±0.9 1.6±0.8 1.5±0.5 * 1.6±0.4 |
Annotate: * and matched group compare, p＜0.05
The labyrinth promotes experiment (anxiety experiment) experimental provision and is made up of four arm districts that are " ten " font, brachium 50 * 50cm, wherein 2 is airtight arm district, the high 40cm of wall, and 2 is the open arms district, there is the high surrounding edge of 1cm at the edge, hoisting depth 50cm, the behavior of mice is observed by the central authorities that then mice put into cross, time is 5 minutes, and record enters the number of times and the time in two kinds of arm districts respectively.Mice has avoidance to enter the open arms district and is confined to the active habit in airtight arm district, if the activity in the open arms district reduces, comprise entering number of times (OE) and the percentage ratio of the time of staying (OT) that the anxiety reaction of prompting mice reduces, and so weighs the anxiety degree of experimental mice.Result's (seeing Table 3) shows that wood frog bioactive peptide component has calm effect antianxity.
Table 3. wood frog bioactive peptide component is to the influence of mouse anti anxiety ability
N=10)
| g/kg·bw | OE％ | OT％ |
| 0.00 0.25 0.50 1.00 | 30.4±1.8 29.1±2.3 26.7±3.6 * 27.9±2.5 * | 32.1±2.5 30.5±3.6 28.3±2.1 * 27.5±4.2 * |
Annotate: * and matched group compare, p＜0.01
Swimming with a load attached to the body experiment (resisting fatigue experiment) is put mice and is swum in the swimming case that has the depth of water greater than 30cm.The load metal object of 5% body weight of Mus root of the tail portion.Beginning when mice sinks to emerging (motion is exhausted) from swimming with mice is the mice swimming time.The result shows that each experimental group swimming time is 246 ± 78,321 ± 93,356 ± 48,402 ± 61, and the experimental group swimming endurance is than the obvious enhancing of matched group (p＜0.01,0.05).
Regulate female endocrine experiment (regulating the estrogen secretion experiment): 40 of 15 month female kunming mices, basic forage feed, be divided into 4 groups at random, give 0.00,0.25,0.50 respectively, the wood frog bioactive peptide component of 1.0g/kgbw irritates stomach 30d, carry out the test of serum estrogen concentrations, the result shows that the control group mice serum estradiol concentration is 4.06 ± 0.78ng/L, each experimental mice serum estradiol concentration is respectively 4.94 ± 0.98,5.07 ± 0.78,5.32 ± 0.12ng/L, more all has significant difference (p＜0.01,0.05) with matched group.Illustrate that the wood frog bioactive peptide can increase the estrogen secretion of experiment mice, have and delay involutional effect.
Enhancing immunity experiment: mice carbon corridor is tested 40 of kunming mices clearly, 23-26g, and basic forage feed is with 0.00,0.25,0.50, the wood frog bioactive peptide continuous irrigation stomach 30d of 1.0g/kg.bw.Every Mus is pressed 0.1mL/10g.bw injects 4 times of dilutions through the tail vein india ink.Inject behind the prepared Chinese ink 2min and 10min and adjoin venous plexus from ophthalmic respectively and get blood 20 μ L, be added to 2mL0.1%Na
2CO
3In the solution, spectrophotometer 600nm wavelength place's photometry density value.The result shows 0.50, the active Toplink of the wood frog of 1.0g/kg.bw dosage strengthens the immunity (table 4) of mice.
Table 4. wood frog bioactive peptide is cleaned up the influence of effect to mice carbon
N=10)
| g/kg·bw | Absorbance | |
| 2min | 10min | |
| 0.00 0.25 0.50 1.00 | 0.18±0.06 0.20±0.01 0.23±0.04 * 0.28±0.03 * | 0.17±0.04 0.19±0.05 0.21±0.02 * 0.25±0.07 * |
Annotate: * and matched group compare, p＜0.05
With wood frog bioactive peptide component is the composition of active components pharmacodynamic experiment
Compositions is formed: wood frog bioactive peptide component 15%, and Semen Caryae Cathayensis 30%, Fructus Cannabis 20%, Semen pruni armeniacae 20%, all the other are water and other excipient etc.Semen Caryae Cathayensis, Fructus Cannabis, Semen pruni armeniacae are pulverized respectively and decocted with the conventional method water, filter to such an extent that liquid concentration becomes clear paste, mix with wood frog bioactive peptide component again.The sample reference composition does not contain wood frog bioactive peptide component.
Effect of relaxing bowel experiment: compositions and sample reference composition dosage are designed to 3.30g/kgbw, continuous irrigation stomach 10d, experiment contrast 0.00g/kg.bw group gives capacity filling stomaches such as distilled water, every group of 20 animals (each 10 of following 2 two experiments), divide cage basic forage feed, do mouse small intestine respectively and advance experiment and the experiment of mice defecation.
Intestinal propulsion experiment mice fasting 16h overnight, tried thing once to each dosage group inferior morning.Give distilled water to each dosage group, matched group behind the 30min.20min irritates stomach once by every Mus 0.2ml to all animals per os with carbon element prepared Chinese ink more at interval.Put to death animal and the extremely ileocecal whole section small intestinal of taking-up stomach of cutting open the belly behind 20min, measuring and advancing the distance in forward position from pylorus to prepared Chinese ink is the prepared Chinese ink advance distance, calculates each treated animal prepared Chinese ink propelling rate.Experimental result shows that experimental group and each matched group compare, and small intestinal prepared Chinese ink propelling rate is obviously rising (P＜0.05) all.
Defecation experiment mice fasting 24h overnight, each group gives compound recipe ground phenol promise ester, and each dosage group gives compositions and sample reference composition behind the 1h, and behind the 20min, every Mus is given carbon element prepared Chinese ink 0.2ml and irritates stomach once.Normal feed water, melena time and 12h defecation total amount arranged first in record.Experimental result shows that experimental mice defecation time shortening first, 12h defecation total amount increase, and compares with the sample matched group with the experiment contrast group to have significant difference (P＜0.01, P＜0.05).
Conclusion: this compositions can obviously promote mouse small intestine prepared Chinese ink propelling rate, can obviously shorten the defecation time first that causes mice with constipation, increases the mice feces volume, has functions of loosening bowel relieving constipation.
Regulate the mouse blood sugar functional experiment: Male Kunming strain mice, basic forage feed, body weight 18-22g, be divided into 4 groups, every group 10, wherein one group is the blank group, induce the hyperglycemia symptom with alloxan (tail vein injection 70 μ g/g.bw every day) for all the other 3 groups, irritate respectively simultaneously that stomach gives 0.00, the compositions of 6.60g/kgbw and the sample reference composition of 6.60g/kgbw, 21d (1h after the administration) wins eyeball and gets blood, detects fasting glucose (glucoseoxidase and peroxidase coupling method).The result shows, above-mentioned sample composition fasting glucose is 7.21 ± 0.42,5.79 ± 0.67mmol/L, sample matched group (6.60g/kgbw) fasting glucose is 6.87 ± 0.58mmol/L, experimental group is compared with experiment contrast group and sample contrast all has significant difference (p＜0.05), the above-mentioned fasting glucose of respectively organizing is all than blank group (4.50 ± 0.98 mmol/L) high (p＜0.05), there was no significant difference between experiment contrast group and sample matched group (p＞0.05).
Adjusting rat fat experiment: get 40 of Wistar rats, male, body weight 230-270g is equally divided into 4 groups at random, except that normal blank group gives the basic feedstuff, all the other respectively organize feedstuff below the equal feed, in basic feedstuff, add 2% and go into cholesterol, 10% Adeps Sus domestica, 0.2% methylthiouracil raised for 6 weeks.Gastric infusion was carried out in the 3rd week in experiment beginning back, was respectively 0.00 every day, the compositions of 6.60g/kgbw dosage and the sample reference composition of 6.60g/kgbw dosage, 21d altogether.Get blood and begin fasting 12h preceding 1 day evening, freely drink water, inferior administration in morning 1 time, the 2h broken end is got blood, blood test cholesterol, triglyceride, high density lipoprotein after administration.The result shows, compares with the blank group, and experimental group cholesterol and content of triglyceride obviously increase, and hdl concentration obviously reduces (p＜0.01).Experimental group and experiment contrast and sample blank group more all can make cholesterol and content of triglyceride obviously reduce, and obviously increase hdl concentration (P＜0.05, p＜0.01) simultaneously, see Table 5.
Table 5. wood frog bioactive peptide compositions to the influence of experimental rat blood fat (mmol/L,
N=10)
| Group (dosage g/kgbw) | Cholesterol | Triglyceride | High density lipoprotein |
| Blank experiment contrast (0.00) sample contrast (6.60) experimental group (6.60) | 3.37±0.87 5.26±0.68 * 5.30±0.75 * 3.98±0.83 *△ | 1.1 3±0.49 2.25±0.39 * 2.09±0.43 * 1.79±0.52 *△ | 3.32±0.32 1.55±0.34 * 1.59±0.50 * 2.08±0.29 *△ |
Annotate: * and blank group compare, p＜0.01; △ and experiment contrast and sample matched group be P＜0.05 or P＜0.01 relatively
It is dietotherapeutic preparation method of composition of active component and uses thereof that the present invention aims to provide with wood frog bioactive peptide component, it is characterized in that with the wood frog being that the wood frog bioactive peptide component of raw material separation and purification or applied molecular biology technology synthetic is an active component, mix the dietotherapeutic compositions that forms with acceptable auxiliary on pharmacy and the Food technology.
Described is the dietotherapeutic compositions of active component with wood frog bioactive peptide component, it is characterized in that described wood frog bioactive peptide component is for being the wood frog bioactive peptide component of raw material separation and purification or applied molecular biology technology synthetic with the wood frog.
Described is the dietotherapeutic compositions of active component with wood frog bioactive peptide component, it is characterized in that described wood frog bioactive peptide component is the 1-40% of composition weight ratio.
Described is the dietotherapeutic compositions of active component with wood frog bioactive peptide component, it is characterized in that said composition is the preparation that is fit to dietotherapeutic.
Described is the dietotherapeutic compositions of active component with wood frog bioactive peptide component, it is characterized in that described suitable dietotherapeutic preparation can be made into injection, oral liquid, drop pill, soft capsule, granule, powder, dispersible tablet, capsule, tablet, chewable tablet, buccal tablet, oral cavity disintegration tablet etc.
Described is the dietotherapeutic compositions of active component with wood frog bioactive peptide component, can be used for slow down aging, improves looks, delays to alleviate climacteric syndrome, prophylactic treatment constipation, resisting fatigue, raising immunity, anxiety, blood fat and blood glucose regulation etc.
Below be specific implementation approach of the present invention, but be not limited in following example.
Embodiment 1
Wood frog bioactive peptide component 5-15%, microcrystalline Cellulose 20-30%, soluble starch 10-25%, silica gel 1-3%, Semen pruni armeniacae (peeling is fried) 10-15%, Fructus Cannabis (stir-fry) 5-10%, all the other are quality improver such as essence, sweeting agent etc.Adopt method commonly used at present to carry out micronize, as high velocity air pulverizer, air-flow vortex pulverizing mill, air-flow micronizer and ball mill, can also adopt spray drying to make powder through being dissolved in solvent, perhaps irritate capsule and become capsule, perhaps compacting in flakes.
Embodiment 2
Wood frog bioactive peptide component 10-30%, Semen Caryae Cathayensis 15-30%, Fructus Cannabis 10-20%, Semen pruni armeniacae 15-25%, all the other are gelatin and other excipient etc.Semen Caryae Cathayensis, Fructus Cannabis, Semen pruni armeniacae are pulverized respectively and decocted with the conventional method water, filter to such an extent that liquid concentration becomes clear paste, mix with wood frog bioactive peptide component, gelatin excipient etc. again, press soft capsule prepared soft capsule.
Embodiment 3
Wood frog bioactive peptide component 5-15%, Semen pruni armeniacae 0-0.1%, Radix Ginseng 0-0.1%, Fructus Jujubae 0.6-0.8%, Auricularia 0.1-0.3%, beans 10-17%, white sugar 5-10%, thickening agent (carboxymethyl cellulose) 1-8%, Fructus Cannabis 1-5%, all the other are animal milk and water.With Fructus Cannabis, Semen pruni armeniacae, Radix Ginseng is raw material, and through flooding twice, merge extractive liquid, concentrates, serosity made from Fructus Jujubae, Auricularia, beans routine and animal milk merge again, become soy milk slurry through conventional pasteurization again, or the spray-dried soy milk powder of making, or conventional tabletting becomes milk slice.
Claims (6)
1. the present invention is characterized in that for being the dietotherapeutic compositions of active component with wood frog bioactive peptide component with active substance wood frog bioactive peptide component be active component, mixes the dietotherapeutic compositions that forms with acceptable auxiliary on pharmacy and the Food technology.
2. described is the dietotherapeutic compositions of active component with wood frog bioactive peptide component, it is characterized in that described wood frog bioactive peptide component is for being the wood frog bioactive peptide component of raw material separation and purification or applied molecular biology technology synthetic with the wood frog.
3. according to claim 1 is the dietotherapeutic compositions of active component with wood frog bioactive peptide component, it is characterized in that described wood frog bioactive peptide component is the 1-40% of composition weight ratio.
4. according to claim 1 is the dietotherapeutic compositions of active component with wood frog bioactive peptide component, it is characterized in that said composition is the preparation that is fit to dietotherapeutic.
5. according to claim 4 is the dietotherapeutic compositions of active component with wood frog bioactive peptide component, it is characterized in that described suitable dietotherapeutic preparation can be made into injection, drop pill, soft capsule, granule, dispersible tablet, capsule, tablet, chewable tablet, buccal tablet, oral cavity disintegration tablet etc.
6. claim 1-5 is described is the dietotherapeutic compositions of active component with wood frog bioactive peptide component, can be used for slow down aging, improves looks, delays to alleviate climacteric syndrome, prophylactic treatment constipation, resisting fatigue, raising immunity, anxiety, blood glucose and blood lipid regulation etc.
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| CNA2006101450063A CN100998865A (en) | 2006-09-21 | 2006-11-29 | Modicine and food dual-purpose composition using forest frog active peptide component as active constituent and its use |
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| CN200610153060 | 2006-09-21 | ||
| CN200610153060.2 | 2006-09-21 | ||
| CNA2006101450063A CN100998865A (en) | 2006-09-21 | 2006-11-29 | Modicine and food dual-purpose composition using forest frog active peptide component as active constituent and its use |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101933614A (en) * | 2010-08-20 | 2011-01-05 | 林慧 | Method for preparing oviductus ranae granules |
| CN101978884A (en) * | 2010-08-23 | 2011-02-23 | 张雪峰 | Stewed Chinese forest frog buccal tablets and preparation method thereof |
-
2006
- 2006-11-29 CN CNA2006101450063A patent/CN100998865A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101933614A (en) * | 2010-08-20 | 2011-01-05 | 林慧 | Method for preparing oviductus ranae granules |
| CN101978884A (en) * | 2010-08-23 | 2011-02-23 | 张雪峰 | Stewed Chinese forest frog buccal tablets and preparation method thereof |
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