CN1478544A - Use of polypeptide protein kind active substance in Arca inflate Roeve and its extraction method - Google Patents
Use of polypeptide protein kind active substance in Arca inflate Roeve and its extraction method Download PDFInfo
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- CN1478544A CN1478544A CNA031397638A CN03139763A CN1478544A CN 1478544 A CN1478544 A CN 1478544A CN A031397638 A CNA031397638 A CN A031397638A CN 03139763 A CN03139763 A CN 03139763A CN 1478544 A CN1478544 A CN 1478544A
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- active substance
- polypeptide protein
- scapharca broughtonii
- class active
- arca
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- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
An application of the polypeptide protein in Arca inflate reeve in preparing antineoplastic medicine is disclosed. A process for extracting said polypeptide includes extracting, chromatography with ion exchange column, desalting, and chromatography with gel column. Said medicine features high curative effect (79.6%) and low poison.
Description
Technical field
The invention discloses a kind of Scapharca broughtonii (Arca inflate Reeve; Scapharcabroughtonii) purposes and the extracting method thereof of polypeptide protein class active substance in.
Background technology
2003 the 2nd phase P37 of " Chinese marine drug " magazine have introduced the proteic separation method of Concha Ostreae.Fresh Concha Ostreae software, with the homogenate of 2 times of amount normal saline, the centrifugal 15min of 3000r/min gets supernatant under 4 ℃, adds respectively (NH)
2SO
4, make (NH) of solution
2SO
4Saturation reach respectively under 50%, 75% and 100%, 4 ℃ and leave standstill 12h.With the centrifugal 15min of 3000r/min, get precipitation again, after the low amounts of water dissolving, pour the bag filter desalination into.Every 2h changes water once.Until 15%CaCl
2Detect in the solution and do not have SO
4 2-Exist, obtain albumen at last.The trophic analysis of Concha Ostreae extract shows, contains abundant sugared source, protein and essential amino acids, particularly content of taurine height in the Concha Ostreae, also contains multiple necessary trace element and multivitamin.Compendium of Material Medica etc. have all been put down in writing the therapeutic efficiency of Concha Ostreae, and Concha Ostreae is used as medicine in the history in existing thousand of China.In addition, the applicant does not find in retrieving about the medicinal report of Scapharca broughtonii polypeptide protein class active substance and the report of extracting method thereof.
(NH) that uses in the method for above-mentioned bibliographical information
2SO
4Settling step extracts albumen, and what obtain is thick total protein, wherein contains plurality of impurities such as polysaccharide, is multiple mixture of ingredients.Extract obtained can not being directly used in the high purposes of purity requirement.
Summary of the invention
Purpose of the present invention is exactly the medical usage that discloses polypeptide protein class active substance in a kind of Scapharca broughtonii, discloses its extraction separation method simultaneously, makes the purity of extract improve greatly, to obtain the higher polypeptide protein class active substance of purity.
The present invention realizes by the following technical solutions.After the cleaning of Scapharca broughtonii content, add distilled water in the certain mass ratio, with 5000r/min homogenate 3min, centrifugal removing slag got supernatant; Supernatant is carried out ion-exchange chromatography, collect each peak respectively after, desalination separates through gel filtration chromatography again, obtains various polypeptide protein class active substances by the molecular weight difference.
With this polypeptide protein class active substance, be equipped with proper additive, make the medicine of certain dosage form, can be used for suppressing tumor.
Antitumor drug with polypeptide protein class active substance of the present invention is made can have 79.60% tumour inhibiting rate, because be the natural biological extraction of substance, has the characteristics of low toxicity simultaneously.The extracting method of Scapharca broughtonii polypeptide protein class active substance of the present invention, directly Scapharca broughtonii albumen is carried out ion-exchange chromatography, after the desalination, carry out gel filtration chromatography again and separate, can reach the effect that the polypeptide protein extract is obtained purification in separation and Extraction, be applicable to medical usage better.
The specific embodiment
The invention will be further described below in conjunction with embodiment.Get Scapharca broughtonii, shell, get its content, after 4 ℃ of ordinary waters cleanings 3 times, the distilled water that reuse is 4 ℃ cleans twice; Add distilled water in certain mass ratio (as 1: 3),, on centrifuge, remove slag, get supernatant behind the centrifugal 10min with 5000r/min homogenate 3min; Supernatant is carried out ion-exchange chromatography, by this step, with extract according to different separation and purification to the Ion Exchange Medium absorbability.After collecting each peak respectively, desalination.Separate through gel filtration chromatography after the desalination, further the extract with identical absorption affinity separates according to different molecular weight again, separates to obtain the protide active substance, and its composition and purity are greatly improved.
Get the interior tumor experiment that above-mentioned protein extract carries out tumor-bearing mice, get male mice in kunming, body weight 19～21g, random packet, 10 every group.Every mice left side oxter inoculation 0.2mlS
180The tumor liquid of ehrlich carcinoma.Begin next day with the negative contrast of normal saline, lumbar injection is carried out in the positive contrast of 5-fluorouracil, injects and weighs after 8 days, dissects the subcutaneous tumors piece, weighs.The curative effect of solid tumor heavily suppresses percentage rate with tumor and represents that computing formula is as follows:
The heavy suppression ratio %=(1-T/C) * 100% of tumor
T: it is heavy that average tumor is organized in treatment
C: the average tumor of negative control group is heavy
Medicine is to mice S
180Before and after heavy (g) tumour inhibiting rate administration of the average tumor of the inhibitory action group dosage route of administration the weight of animals (g) of tumor
And (%) body weight change normal saline 20ml/kg ip * 8 20.74 23.65 1.61 sample 250mg/kg ip * 8 20.23 17.47 0.33 79.60-13.64%5-fluorouracil 10ml/kg ip * 8 19.96 15.3 0.377 76.61-23.35% behind the natural law medicine prodrug
Experimental result shows that the tumour inhibiting rate of the activated protein that the present invention extracts is up to 79.60%, and its toxicity is lower than positive control 5-fluorouracil group (weight loss is starkly lower than the 5-fluorouracil group before and after the sample sets mouse experiment, illustrates that its toxicity is lower); Simultaneously, said preparation also has high inhibitory to the cultivation of tumor cell in vitro.Therefore, characteristics such as extract of the present invention has high-efficiency low-toxicity, and manufacturing process is pollution-free, being expected to provides a kind of safe, effective, quality controllable new type antineoplastic medicine for clinical.
Claims (3)
1, a kind of Scapharca broughtonii (Arca inflate Reeve; Scapharca broughtonii) purposes of polypeptide protein class active substance in: it is characterized in that this polypeptide protein class active substance is used to prepare the medicine that suppresses tumor.
2, a kind of Scapharca broughtonii (Arca inflate Reeve; Scapharca broughtonii) extracting method of polypeptide protein class active substance in is characterized in that: after the Scapharca broughtonii content is cleaned, add distilled water in the certain mass ratio, with 5000r/min homogenate 3min, remove slag, get supernatant; Supernatant is carried out ion-exchange chromatography; After collecting each peak respectively, desalination; Separate through gel filtration chromatography again, obtain polypeptide protein class active substance.
3, by the extracting method of polypeptide protein class active substance in the described Scapharca broughtonii of claim 2, it is characterized in that: the content cleaning step of described Scapharca broughtonii is: after cleaning 3 times with 4 ℃ of ordinary waters earlier, the distilled water that reuse is 4 ℃ cleans twice; The mass ratio that described distilled water adds is 1: 3.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA031397638A CN1478544A (en) | 2003-07-09 | 2003-07-09 | Use of polypeptide protein kind active substance in Arca inflate Roeve and its extraction method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA031397638A CN1478544A (en) | 2003-07-09 | 2003-07-09 | Use of polypeptide protein kind active substance in Arca inflate Roeve and its extraction method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1478544A true CN1478544A (en) | 2004-03-03 |
Family
ID=34155085
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA031397638A Pending CN1478544A (en) | 2003-07-09 | 2003-07-09 | Use of polypeptide protein kind active substance in Arca inflate Roeve and its extraction method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1478544A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103739690A (en) * | 2013-11-29 | 2014-04-23 | 于荣敏 | Method for separation preparation of anti-tumor polypeptide compound from ArcainflataReeve, and uses of anti-tumor polypeptide compound |
| CN117802055A (en) * | 2024-01-03 | 2024-04-02 | 暨南大学 | Deoxyarteannuin B synthase and application thereof |
-
2003
- 2003-07-09 CN CNA031397638A patent/CN1478544A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103739690A (en) * | 2013-11-29 | 2014-04-23 | 于荣敏 | Method for separation preparation of anti-tumor polypeptide compound from ArcainflataReeve, and uses of anti-tumor polypeptide compound |
| CN117802055A (en) * | 2024-01-03 | 2024-04-02 | 暨南大学 | Deoxyarteannuin B synthase and application thereof |
| CN117802055B (en) * | 2024-01-03 | 2024-06-11 | 暨南大学 | Deoxyarteannuin B synthase and application thereof |
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