CN1163564A - Enterogenin pharmaceutical form - Google Patents
Enterogenin pharmaceutical form Download PDFInfo
- Publication number
- CN1163564A CN1163564A CN 95195713 CN95195713A CN1163564A CN 1163564 A CN1163564 A CN 1163564A CN 95195713 CN95195713 CN 95195713 CN 95195713 A CN95195713 A CN 95195713A CN 1163564 A CN1163564 A CN 1163564A
- Authority
- CN
- China
- Prior art keywords
- intestinal
- aglycon
- ampoule
- powder
- preparation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention relates to the obtaining of two pharmaceutical forms-pulvis and ampules of the veterinary medical preparation ''Enterogenin''. The bioactive ingredient of the preparation is isolated by fractionation of pig small intestinal mucosa cell, extraction with acetic acid, sedimentation with ethyl alcohol, ion-exchange chromatography and diaflo ultrafiltration in the range of 0.5-2 kDa. The bioactivity of the preparation is related to two nucleopeptides: guanosine tripeptide and adenosine heptapeptide. The basic criterion for the biological action of ''Enterogenin'' pulvis and ampules is the activation of biosynthesis of nucleic acids and specific proteins in the muscles, liver, intestines and spleen.
Description
The present invention relates to the separation method of " intestinal aglycon-powder and ampoule " preparation, its industrial level, the production method of pharmaceutical dosage form and the research method of said preparation pharmacotoxicological effect.The active component of " intestinal aglycon-powder and ampoule " is from the isolating bioactive substance of intestinal mucosa.Existing bibliographical information exists the material of propagation capable of inhibiting cell (1,2,3) in intestinal mucosa.There is not report that the separation of the material of generation stimulation can take place form in the document.We had once applied for a kind of method (4,5) of the intestinal mucosa separating bio pungent from homoiothermic animal.This method exists some shortcomings: complicated technical process makes the production cost of this material relatively costly, and this method can not be produced the dosage form of final pharmacology test.
The objective of the invention is: the method for 1) improving the bioactive ingredients of described separation " intestinal aglycon-powder and ampoule " preparation; 2) pharmaceutical dosage form of development " intestinal aglycon-powder and ampoule " preparation; 3) effect of bioactive ingredients is being carried out after the experimental research pharmacotoxicological effect of test " intestinal aglycon-powder and ampoule " preparation in the practice of veterinary and domestication of animals.
First purpose is finished by the method for separating bio building-up process stimulator from the intestinal mucosa of pig.Production decision mainly may further comprise the steps: 1) separate specific cell mass from the animal intestinal mucosa, this intestinal mucosa is the garbage between the casing cleaning Vehicle of meat Package factory; 2) alcohol acid extraction thing will be obtained after the heavy polymer sedimentation; In this step, should prevent the relevant pollution of any and initial Biomass; 3) carry out the ion exchange fractionated in batches; 4) eluent that fractionated is obtained carries out ultra-filtration.Technical scheme is described in embodiment 1.1.Compare with the method for putting down in writing among the patent BG49927MPK A61K37/02, have following improvement: the consumption of reagent is less; Some steps in the basic technical scheme are changed, reduced number of steps; The parameter of ultra-filtration has been carried out change and carried out further definite.Consequently, isolated two kinds rather than a kind of biological activity nuclear peptide (nucleopeptide), we are with its called after " zest enterocyte aglycon ".
Second purpose finished by preparing two kinds of pharmaceutical dosage forms: the enteric coated particles packing that 1) is used for " the intestinal aglycon powder " of oral administration; 2) contain purification the deposit form biologically active agents " intestinal aglycon ampoule " bottle.When preserving for 2 to 6 ℃, this pharmaceutical dosage form can be in 3 years effectively.The analysis of said medicine dosage form has been obtained the official confirmation (6) of veterinary formulation control laboratory and national veterinary drug office.This analysis comprises the identification of the chemistry and biology of the biological activity nuclear peptide in " intestinal aglycon ".In addition, give the analytical method of carrier components in the preparation.
The 3rd purpose chamber and clinical experiment and should be used in veterinary's practice by experiment finished.Biological activity nuclear peptide contained in " intestinal aglycon-powder and ampoule " works at molecular level; They make regenerated shortening proliferating cycle of physiological of intestinal mucosa cells, thereby cause the increase of small intestinal sorbent surface; For nonproliferating cell, their stimulate the synthetic of specified protein: separate toxalbumin in the actomyosin in the striped muscle, liver, cause immune protein in the spleen; And the digestive enzyme in the gastrointestinal tract.In smooth muscle cell, these nuclear peptides are by stimulating Ca
2+-flow into and induce depolarization (7).
In little laboratory animal, use every day weight increase that said preparation can make them more than 200%.Use the secular effect of stimulation that " intestinal aglycon " can produce lasting six months in 20 days pig and cattle breeding field.The characteristics of this effect are, can make the weight of pig increase 125g every day, and cattle increases 110g every day.For chicken, made an addition to the drinking water in the 3rd day to the 45th day after hatching, " intestinal aglycon " stimulates DNA and proteinic biosynthesis, makes their weight increase 118%." intestinal aglycon " also shows therapeutic effect in animal model: in experimental gastric ulcer, after the liver poisoning, come prophylaxis against infection diseases by immune stimulatory.Through national tumor center and national drug management research test confirm " intestinal aglycon " nontoxic, no teratogenesis, nontoxic, and hereditary-less toxicity to the embryo.
The present invention will be described by following examples now.
Embodiment 1. tasks 1 (Fig. 1). on the biterling machine in slaughterhouse's casing cleaning shop, the material of the enrichment of cell that collection is extruded from No. 3 casing squeegee rollers, and the cold acetic acid of while adding 20% under constant stirring, the ultimate density that makes acetic acid is 2% (with respect to cumulative volume).Processing procedure to Biomass can be proceeded, and also Biomass can be stored in-40 ℃ the refrigerator.If take a kind of method in back, need to carry out partially thawed subsequently.Then handle with meat grinder.The ethanol (0.2 times of volume of total amount) of adding 95% in the material that rubs.Then Biomass is heated to 70 ℃ rapidly and also is cooled to room temperature immediately.Tell liquid extraction liquid by separation and filtration.Discard high-molecular weight chip.Acid ph value is transferred to 6.5-7.0.Alcohol and lipid mixture are transferred in the ether of glass reactor.Be adsorbed onto extract on the DEAE cellulose under the constant salinity and using base brine solution eluting in batches.Activated part is carried out purification by penetrating amicon, collects the part in the 0.5-2.0kDa scope.Filter for the second time and under aseptic condition, carry out.Measure the concentration of nuclear peptide, this concentration is essential by the dosage of determining pharmaceutical dosage form.The preparation of embodiment 1. tasks 2. " intestinal aglycon-powder and ampoule " pharmaceutical dosage form is by carrying out for the designed chemistry and biology method of evaluation nuclear peptide: a) ultra-violet absorption spectrum-maximum 260nm, minima 237nm; A
260/ A
237=1.21-1.19; B) characteristic reaction of peptide, for example, the ninhydrin reaction of Reichelt; C) mensuration of molecular weight: by using the Sephadex G-25 buttress shaft of crossing by the known peptide calibration in advance of molecular weight; For K
Av, the reference limit of two kinds of active nucleus peptides is respectively 1.1-1.3kDa and 0.6-0.65kDa; D) aminoacid is formed: analyze by " fingerprint " on the paper chromatography, or more accurately, by the quantitative assay on amino-acid analyzer.In low-molecular-weight nuclear peptide, glycine and tryptophan have been recorded; Contain arginine, serine, leucine, histidine, glycine and tyrosine in another nuclear peptide; E) quantitative analysis: spectrophotometric analysis A
260, be contrast with the 0.01mmol adenine sulfate of standard; Computing formula:
F) biological activity is by measuring
3The incorporation of H-leucine (peritoneal injection, dosage are 0.04MBp/20g mice body weight) is measured.Add 0.02mg " intestinal aglycon " (experimental group) after 18 hours at injection isotope (matched group) and isotope, measure the isotopic incorporation of taking from the Adductorius 50mg muscular tissue of mice back leg.
Flow chart
The separation of zest intestinal aglycon
Embodiment 1. tasks 3. are to measure DNA, RNA and proteinic biosynthesis by the incorporation of corresponding isotope-labeled precursor to the rapid evaluation methodology of the biologic activity of " intestinal aglycon-powder and ampoule " preparation.Average activation is greater than 200% (table 1).With occurring secular morphological change (table 2) in the animal of " intestinal aglycon " repeated treatments.The sorbent surface of intestinal increases, thereby makes the usefulness of food increase by 2.56 times when the picked-up small amounts of food, and makes proteinic usefulness coefficient ratio matched group increase by 2.5 times.Morphologic change has also taken place in striped muscle, and compressor protein increases relatively.To 6 said preparations of calf injection; Added said preparation 20 days every day in the food of pig and fresh-water fishes; And in the drinking water of chicken, added said preparation 45 days, their growth kinetics is improved.
In rat, observe " intestinal aglycon " notable therapeutic effect to two kinds of experimental ulcer models (reserpine and anxiety-histamine model).The metabolism of hepatorenal syndrome and the kinetics of recovery process after the rat carbon tetrachloride poisoning, have been studied." intestinal aglycon " preparation shows protection and repair.
Table 1 " intestinal aglycon " to DNA (
3The incorporation of H-thymus pyrimidine), RNA (
14The incorporation of C-uracil) and protein (
3The leucic incorporation of H-) biosynthetic influence [with respect to the percent of the control animals that does not give " intestinal aglycon "]
| Animal | Organ | Incorporation | ||
| Thymus pyrimidine | Uracil | Leucine | ||
| Rat | Duodenum jejunum small intestinal stage casing ileum liver cell nuclear | ????221 ????190 ????284 ????337 ????16 | ||
| Mice | Duodenum jejunum small intestinal stage casing ileum muscle | ????163 ????302 ????212 ????153 | ????360 ????160 | ????212 ????117 |
| Chicken | White muscle small intestinal stage casing red muscle liver | ????103 ????242 ????118 ????183 | ????104 ????265 ????193 ????331 | |
Morphological change [with respect to the percent of the control animals that does not give " intestinal aglycon "] after table 2 is repeatedly handled with " intestinal aglycon "
| Parameter | Organ | Animal | Change % |
| Body weight (g) | Whole body weight liver small intestinal heart spleen kidney | Mice rat rat rat rat rat | ????212 ????159 ????126 ????135 ????112 ????120 |
| Cellularity (* 10 5/lμm 2) intestinal mucosa | Ileum duodenum jejunum small intestine stage casing, duodenum jejunum small intestine stage casing ileum | Rat rat rat rat mouse mouse mouse mouse | ????144 ????173 ????167 ????135 ????134 ????163 ????173 ????139 |
| The quantity of intestinal villus (microscopically section) | The small intestinal stage casing | Mice | ????133 |
| The length of intestinal villus (microscopically section/μ m) | The small intestinal stage casing | Mice | ????125 |
| Single myofibrillar area (μ m 2) | Gastrocnemius | Pig | ????191 |
| Diaphragm | Pig | ????279 | |
| Oblique | Pig | ????252 | |
| Single myofibrillar diameter (μ m is under the ultramicroscope) | Gastrocnemius | Pig | ????154 |
| Diaphragm | Pig | ????170 | |
| Oblique | Pig | ????157 | |
| The thickness of Z stricture of vagina (μ m is under the ultramicroscope) | Gastrocnemius | Pig | ????300 |
List of references
1.Skraastad 0, Reicelt KI. endogenous colon mitotic inhibitor reduces colon cancer cell (HT29) hypertrophy in the serum finite medium.Virchows?Arch?B，50，(1989)，393-390。
2.Skraastad 0, Reichelt KI. endogenous colon mitotic inhibitor and edible calcium suppress by the inductive colon cell hypertrophy of cholic acid.J?Gastroent?23(1986)，801-807。
3.Triffonov B, Roussev GK, Argirov C, Draganov M, the 3T3 mouse fibroblast cell that the biological action of a Kamberov B. new intestinal peptide-inhibition is cultivated and the enterocyte aglucon of L5178Y mouse lymphotactin.Regulatory?Peptides，51(1994)，111-119。
4.Triffonov B, Roussev GK, Boshev H, the method for Tyanev G. separating bio stimulus object.Patent?49927，BG?49927?MPK?A61K37/02，Vol?3，March16，1992，1-3(Bulgarian)。
5.Roussev GK, Triffonov B, Petrov M, boshev H. separates the method that the shaping active substance is arranged.Invention?patent?37396?MPK-A61K35/38，Vol?6，June14，1985，1-6。
6. certificate number XIII-33/10.02.1995, the veterinary of country of the Ministry of Agriculture does the permission of permission at veterinary industry production and use " intestinal aglucon " preparation.
7.Triffonov B, Kristev A, Zaprianov G, Lukanov J, a KostadinovaI. new intestinal peptide (intestinal aglucon) is to the contraction of rat and Cavia porcellus intestinal smooth muscle and the effect of bio-electrical activity.J?Gastrointest?Motil?4，(1992)，193-l99。
Claims (3)
1, the pharmaceutical dosage form of biosynthetic process stimulator, comprise the biological activity nuclear peptide that contains 0.0125% deposit form and 0.2% the sodium alginate normal saline solution " intestinal aglycon ampoule " bottle and contain 0.125% be adsorbed on the starch and with " the intestinal aglycon powder " of the biological activity nuclear peptide of aquation lipogenesis enteric coated particles.
2, the method for the bioactive ingredients in the dosage form of production claim 1, comprise the steps: depleted intestinal Biomass is carried out fractionated, uses acetic acid extraction, uses the ultra-filtration in ethanol sedimentation, ion exchange chromatography and the 0.5-2.0kDa scope of low concentration, separate obtaining two kinds of biological activitys nuclear peptides.
3, " intestinal aglycon-powder and ampoule " application in veterinary practice: a) for pig, be that " the intestinal aglycon powder " of 100g/200kg body weight/day mixes with feedstuff, used 20-30 days with dosage; B) for ruminant, every " the intestinal aglycon ampoule " of three days intramuscular injection 0.5ml/100kg body weight, totally 6 to 8 times; C) for chicken, every day, " the intestinal aglycon ampoule " with the 0.5ml/10kg body weight mixed totally 45 days with drinking water; D), in feeding period, 100g " intestinal aglycon powder " is mixed with the 5kg feedstuff for fresh-water fishes.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 95195713 CN1163564A (en) | 1995-07-19 | 1995-10-09 | Enterogenin pharmaceutical form |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| BG99795 | 1995-07-19 | ||
| CN 95195713 CN1163564A (en) | 1995-07-19 | 1995-10-09 | Enterogenin pharmaceutical form |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1163564A true CN1163564A (en) | 1997-10-29 |
Family
ID=5082951
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 95195713 Pending CN1163564A (en) | 1995-07-19 | 1995-10-09 | Enterogenin pharmaceutical form |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1163564A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11479693B2 (en) | 2018-05-03 | 2022-10-25 | Avery Dennison Corporation | Adhesive laminates and method for making adhesive laminates |
-
1995
- 1995-10-09 CN CN 95195713 patent/CN1163564A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11479693B2 (en) | 2018-05-03 | 2022-10-25 | Avery Dennison Corporation | Adhesive laminates and method for making adhesive laminates |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP5128760B2 (en) | Γ-polyglutamic acid (γ-PGA, H form) and γ-polyglutamate for use as dietary supplements in dietary products | |
| Jankowski et al. | The effect of different dietary ratios of arginine, methionine, and lysine on the performance, carcass traits, and immune status of turkeys | |
| CN107404903A (en) | Improve the method and composition of the food efficiency of animal such as dog using fresh meat or frozen meat | |
| Janczyk et al. | Evaluation of nutritional value and safety of the green microalgae Chlorella vulgaris treated with novel processing methods | |
| Hempe et al. | Intestinal metallothionein gene expression and zinc absorption in rats are zinc-responsive but refractory to dexamethasone and interleukin 1α | |
| CN103826652A (en) | Method of use of activated functional proteins to improve animal health | |
| CN1330517A (en) | Food-induced antisecretory proteins in egg yolk | |
| CN111718974B (en) | Application of mung bean peptide in preparation of medicine for promoting lead excretion, active peptide and application thereof | |
| CN101309695A (en) | A method for preparing pig placental extract to use as a feed additive and use thereof | |
| Xu et al. | Alfalfa silage diet improves meat quality by remodeling the intestinal microbes of fattening pigs | |
| CN104247859A (en) | Mulberry-leaf additive for pig forage | |
| CN1163564A (en) | Enterogenin pharmaceutical form | |
| Shaposhnikov et al. | Influence of L-lysine sulfate on containing of vitamins and minerals in the body of broiler chickens | |
| Li et al. | The effect of dietary addition of a polysaccharide from Atractylodes macrophala Koidz on growth performance, immunoglobulin concentration and IL-1β expression in weaned piglets | |
| OKUMURA et al. | Incorporation of 15N-urea in chicks | |
| Estornell et al. | Protein synthesis in vivo in rats fed on lipid-rich liquid diets | |
| US5948442A (en) | Stimulating ENTEROGENIN compositions and methods for their isolation and use | |
| CN105941912A (en) | Application of alpha-ketoglutaric acid and alpha-ketoglutarate in preparation of feed additive capable of improving pig carcass composition | |
| CN105558350A (en) | Feed containing Chinese herbal medicine compound crude polysaccharide and preparation method of feed | |
| Slozhenkina et al. | Metrological aspects of using probiotics | |
| CN109497287A (en) | A kind of feed addictive can promote growth of animal | |
| CN101785793A (en) | Application of antradiacomphora extract capsule for protecting liver and promoting body immune regulation function | |
| CN109452226A (en) | A method of improving methionine content in weanling pig serum | |
| Kim et al. | Development and application of a multi-step porcine in vitro system to evaluate feedstuffs and feed additives for their efficacy in nutrient digestion, digesta characteristics, and intestinal immune responses | |
| CN107814836A (en) | A kind of biologically active polypeptide FPGPIPNS and its preparation method and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C53 | Correction of patent for invention or patent application | ||
| CB02 | Change of applicant information |
Applicant after: Alexander A Nova Joe Applicant before: C A. Alexandrov. |
|
| COR | Change of bibliographic data |
Free format text: CORRECT: APPLICANT; FROM: C A ALAIKESANGRAUF TO: JURGEN GELUOYIBEIER |
|
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |