CN101062062A - Method for preparing anti-cancer effective component of marine animals - Google Patents
Method for preparing anti-cancer effective component of marine animals Download PDFInfo
- Publication number
- CN101062062A CN101062062A CNA2007100224695A CN200710022469A CN101062062A CN 101062062 A CN101062062 A CN 101062062A CN A2007100224695 A CNA2007100224695 A CN A2007100224695A CN 200710022469 A CN200710022469 A CN 200710022469A CN 101062062 A CN101062062 A CN 101062062A
- Authority
- CN
- China
- Prior art keywords
- alcoholic solution
- eluent
- extractum
- sample
- effective component
- 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 Containing Plant Substances (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses a preparing method of antineoplastic effective component of sea animal, which comprises the following steps: choosing stone larva as raw material; fore-treating medicinal materials; adding into 30%-60% alcohol solution; stirring for several minuet; hypersonic-extracting for 30-60min; filtering; adding 30%-60% alcohol solution; hypersonic-extracting for 30-60min; cold-soaking for 10-12h; leaching extract; washing residue for 2-3 times with 30%-60% alcohol solution; merging twice extract and washing liquid; centrifuging for 3-5 min with centrifugal machine; reclaiming the alcohol solution with supernatant fluid; condensing; getting concrete; diluting the concrete with water; eluting with up macroporous resin; keeping the sample for 1-2h; proceeding elution condensation. This invention possesses manifest antineoplastic curative effect.
Description
Technical field
The present invention relates to the preparation method of anti-cancer effective component, refer in particular to the preparation method of the anti-cancer effective component of a kind of marine animal.
Background technology
Tumor has now become the healthy and disease more widely of serious threat human life, and there are New Development malignant tumor 9,000,000 people in the whole world every year, and 7,000,000 people that reach that die from this disease.In state-owned 1,300,000,000 populations, annual nearly 1,600,000 New Development malignant tumor patients, annual nearly 1,300,000 people die from cancer, so cancer is positive excess, and to cross that cardiovascular and cerebrovascular disease forms be first cause of death.The control of tumor has become the problem of extensively paying attention in the world wide, countries in the world have been dropped into a large amount of human and material resources and studied every year, but up to now, still not fully aware of to its cause of disease and pathogeny, Therapeutic Method does not still have important breakthrough, and many people have begun sight is turned to wide ocean.Marine organisms have the special living environment that is different from terrestrial life, are wherein containing a large amount of novel structures, special, the extremely intensive material of biological activity of chemical property, have shown great potential at anti-tumor aspect.Research at present mainly concentrate on Mollusca (Jiang Qingfeng, Zhou Youjun, Wang Jinzheng. ocean antitumor drug progress .[J] Chinese marine drug, 2004, (06) .) and seaweeds (patent CN1690076A), antitumor mechanism and action effect have nothing in common with each other.
Mitellae Pollicipes mitella (Linnaeus, 1758) claims the Testudinis foot again, belongs to Arthropoda (Arthropoda) Crustachia (Crustacea) Cirripedia (Cirripedia).All on the books in Compendium of Material Medica, " detailed outline is picked up any lost article from the road ", " language is separated in Hainan ".Its property is flat, sweet in the mouth, and is nontoxic, has effects such as tonification, diuresis, removing food stagnancy, to the also expansion of research of anti-tumor aspect.
Summary of the invention
Order of the present invention is to find a kind of marine animal to have the preparation method of the active component of antitumor action.
Described preparation method is: be raw material with the Mitellae, through the processing procedures such as separation of medical material pre-treatment, extractum extraction, effective ingredient.Be specially:
(1) medical material pre-treatment: get Mitellae and thaw, clean, get meat one by one, smash refiner to pieces with tissue and handle 5-8min, the elimination supernatant gets the pre-treatment medical material;
(2) extractum extracts: the pre-treatment medical material is added the 30%-60% alcoholic solution, and stirred for several minute, supersound extraction 30-60min filters, and extracting solution is standby.Filtering medium adds the 30%-60% alcoholic solution again, supersound extraction 30-60min, and merceration 10-12h leaches extracting solution, and, merge extracted twice liquid and cleaning mixture, the centrifugal 3-5min of usefulness centrifuge with 30%-60% alcoholic solution washing residue 2-3 time, get supernatant and reclaim alcoholic solution, concentrate, get extractum.
(3) separation of effective ingredient: behind extractum dilute with water (the 7-10 times of water yield), last macroporous resin column eluting, keeping sample 1-2h behind the last sample.Carry out eluting with pure water, 30% alcoholic solution, 70% alcoholic solution successively, treat that eluent is limpid, use down a kind of eluent instead behind the odorlessness, collect respectively to merge and respectively organize eluent, concentrate, get final product.
Advantage of the present invention is:
1, found a kind of marine animal with antitumor action: present tumor disease becomes gradually to one of disease of the threat maximum of human health, and it does not find the method for fine this disease of treatment as yet, and constantly seek new species, new chemical compound is resisted tumor.The present invention has found a kind of marine animal with antitumor action---Mitellae.
2, found the preparation method of this marine animal antineoplastic effective composition:, found the preparation method of Mitellae extract anti-tumor active ingredient through the anti-tumor biological screening test.
The specific embodiment
Embodiment one:
Get Mitellae and thaw, clean, get meat one by one, smash refiner to pieces with tissue and handle 5min, the elimination supernatant gets the pre-treatment medical material.Get 200g pre-treatment medical material and add 30% alcoholic solution 300ml, stir 1min, supersound extraction 30min filters, and extracting solution is standby.Filtering medium adds 30% alcoholic solution 200ml again, supersound extraction 30min, and merceration 10h leaches extracting solution, and, merge extracted twice liquid and cleaning mixture with 30% alcoholic solution washing residue 2 times, (2500 change/min), get supernatant with the centrifugal 3min of centrifuge, reclaim alcoholic solution, concentrate, get extractum 5g.
After the dilution of 5g extractum usefulness 50ml water, last macroporous resin column eluting, keeping sample 1h behind the last sample.Earlier use the pure water eluting, be eluted to limpid, the odorlessness of eluent, collect eluent 400ml altogether, be concentrated into 10ml, get sample A1.Reuse 30% alcoholic solution eluting is eluted to limpid, the odorlessness of eluent, collects eluent 280ml altogether, is concentrated into 10ml, gets sample B1.Reuse 70% alcoholic solution eluting is eluted to limpid, the odorlessness of eluent, collects eluent 250ml altogether, is concentrated into 10ml, gets sample C1.Use the ethanol solution eluting at last, be eluted to limpid, the odorlessness of eluent, collect eluent 250ml altogether, be concentrated into 10ml, concentrated solution is pure clear, gives up through the no solute event of thin layer chromatography point sample proof.
Embodiment two:
Get Mitellae and thaw, clean, get meat one by one, smash refiner to pieces with tissue and handle 6min, the elimination supernatant gets the pre-treatment medical material.Get 200g pre-treatment medical material and add 40% alcoholic solution 300ml, stir 1min, supersound extraction 40min filters, and extracting solution is standby.Filtering medium adds 40% alcoholic solution 200ml again, supersound extraction 40min, and merceration 12h leaches extracting solution, and, merge extracted twice liquid and cleaning mixture with 40% alcoholic solution washing residue 2 times, (2500 change/min), get supernatant with the centrifugal 5min of centrifuge, reclaim alcoholic solution, concentrate, get extractum 8g.
After the dilution of 8g extractum usefulness 60ml water, last macroporous resin column eluting, keeping sample 1h behind the last sample.Earlier use the pure water eluting, be eluted to limpid, the odorlessness of eluent, collect eluent 450ml altogether, be concentrated into 10ml, get sample A2.Reuse 30% alcoholic solution eluting is eluted to limpid, the odorlessness of eluent, collects eluent 300ml altogether, is concentrated into 10ml, gets sample B2.Reuse 70% alcoholic solution eluting is eluted to limpid, the odorlessness of eluent, collects eluent 300ml altogether, is concentrated into 10ml, gets sample C2.Use the ethanol solution eluting at last, be eluted to limpid, the odorlessness of eluent, collect eluent 300ml altogether, be concentrated into 10ml, concentrated solution is pure clear, gives up through the no solute event of thin layer chromatography point sample proof.
Embodiment three:
Get Mitellae and thaw, clean, get meat one by one, smash refiner to pieces with tissue and handle 8min, the elimination supernatant gets the pre-treatment medical material.Get 200g pre-treatment medical material and add 60% alcoholic solution 300ml, stir 1min, supersound extraction 60min filters, and extracting solution is standby.Filtering medium adds 60% alcoholic solution 200ml again, supersound extraction 60min, and merceration 12h leaches extracting solution, and, merge extracted twice liquid and cleaning mixture with 40% alcoholic solution washing residue 3 times, (2500 change/min), get supernatant with the centrifugal 5min of centrifuge, reclaim alcoholic solution, concentrate, get extractum 6g.
After the dilution of 6g extractum usefulness 60ml water, last macroporous resin column eluting, keeping sample 1h behind the last sample.Earlier use the pure water eluting, be eluted to limpid, the odorlessness of eluent, collect eluent 450ml altogether, be concentrated into 10ml, get sample A3.Reuse 30% alcoholic solution eluting is eluted to limpid, the odorlessness of eluent, collects eluent 300ml altogether, is concentrated into 10ml, gets sample B3.Reuse 70% alcoholic solution eluting is eluted to limpid, the odorlessness of eluent, collects eluent 300ml altogether, is concentrated into 10ml, gets sample C3.Use the ethanol solution eluting at last, be eluted to limpid, the odorlessness of eluent, collect eluent 300ml altogether, be concentrated into 10ml, concentrated solution is pure clear, gives up through the no solute event of thin layer chromatography point sample proof.
Embodiment four:
The anti-tumor biological screening test:
1, screening technique: tetrazolium (microculture tetrozolium, MTT) reducing process
2, operating procedure
The every hole of (1) 96 orifice plate adds single cell suspension 90 μ l.Blank group only adds complete culture medium, does not contain cell suspension.Behind the cell inoculation, in 37 ℃, 5%CO
2Incubator is cultivated 24h.
(2) positive drug (methotrexate) and testing compound respectively are made into 5 Concentraton gradient (5 times of dilutions), every hole adds 10 μ l, each concentration establish four parallel; Negative control group adds normal saline.Continuous culture 48h.The 5mg/ml MTT solution (Sigma, normal saline preparation) that every then hole adds 20 μ l continues to cultivate 4h.
(3) every hole adds three liquid (the 10%SDS-5% isobutanol-0.1%HCl), spend the night in 37 ℃ of incubators of 50 μ l.
(4) measure the OD value in the 570nm place with microplate reader.
(5) calculate suppression ratio according to following formula, ask IC
50
3, sample and cell strain
Reagent: methotrexate, sample A1, A2, A3 (sample that the pure water eluting obtains), sample B1, B2, B3 (sample that 30% pure eluting obtains), sample C1, C2, C3 (sample that 70% pure eluting obtains).
Cell strain: human leukemia cell line CCRF-CEM, lung cancer cell line A549.
4, result of the test
| IC 50(μg/ml) | Embodiment one sample | Embodiment two samples | Embodiment example three samples | ||||||
| A1 | B1 | C1 | A2 | B2 | C2 | A3 | B3 | C3 | |
| CCRF-CEM A549 | - 102.8 | 94.6 90.5 | 36.2 33.1 | 101.2 98.6 | 92.8 84.4 | 35.1 32.8 | 102.0 98.7 | 96.3 86.2 | 36.7 33.0 |
Reference substance: methotrexate is to the IC of human leukemia cell line CCRF-CEM and lung cancer cell line A549
50Be respectively 0.41 μ g/ml and 0.29 μ g/ml
5, conclusion
In three kinds of embodiment, the IC of Mitellae extract C 1, C2, C3 component
50All be better than A and B component respectively, and embodiment two resulting extractum are maximum, the C2 component is also the highest to the growth inhibitory activity of human leukemia cell line CCRF-CEM and human lung carcinoma cell line A549.
Claims (2)
1. the preparation method of the anti-cancer effective component of a marine animal is characterized in that: be raw material with the SHIJIE, through medical material pre-treatment, extractum extract, the isolating processing procedure of effective ingredient and prepare anti-cancer effective component.
2. the preparation method of the anti-cancer effective component of a kind of marine animal according to claim 1 is characterized in that:
(1) medical material pre-treatment: get SHIJIE and thaw, clean, get meat one by one, smash refiner to pieces with tissue and handle 5-8min, the elimination supernatant gets the pre-treatment medical material;
(2) extractum extracts: the pre-treatment medical material is added the 30%-60% alcoholic solution, stir, supersound extraction 30-60min filters, and extracting solution is standby; The material that obtains after the filtration adds the 30%-60% alcoholic solution again, supersound extraction 30-60min, merceration 10-12h, leach extracting solution, and wash residue 2-3 time, merge extracted twice liquid and cleaning mixture with the 30%-60% alcoholic solution, with the centrifugal 3-5min of centrifuge, get supernatant and reclaim alcoholic solution, concentrate, get extractum;
(3) separation of effective ingredient: after extractum dilute with water 7-10 times, last macroporous resin column eluting, keeping sample 1-2h behind the last sample; Carry out eluting with pure water, 30% alcoholic solution, 70% alcoholic solution successively, treat that eluent is limpid, use down a kind of eluent instead behind the odorlessness, collect respectively to merge and respectively organize eluent, concentrate, get final product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007100224695A CN101062062A (en) | 2007-05-21 | 2007-05-21 | Method for preparing anti-cancer effective component of marine animals |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007100224695A CN101062062A (en) | 2007-05-21 | 2007-05-21 | Method for preparing anti-cancer effective component of marine animals |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101062062A true CN101062062A (en) | 2007-10-31 |
Family
ID=38963476
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2007100224695A Pending CN101062062A (en) | 2007-05-21 | 2007-05-21 | Method for preparing anti-cancer effective component of marine animals |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101062062A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107375338A (en) * | 2017-07-31 | 2017-11-24 | 中国医学科学院药用植物研究所海南分所 | A kind of preparation method and application of Onchidium struma extract |
-
2007
- 2007-05-21 CN CNA2007100224695A patent/CN101062062A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107375338A (en) * | 2017-07-31 | 2017-11-24 | 中国医学科学院药用植物研究所海南分所 | A kind of preparation method and application of Onchidium struma extract |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101747446B (en) | Method for extracting anti-fatigue acidic ginseng polysaccharide | |
| CN101049328A (en) | Technique for producing extractive of olive growing leaves | |
| CN1246325C (en) | Preparation method of hongjintian effective component extract | |
| CN1129572C (en) | Preparation and application of tanshinpolyphenolic salt | |
| CN109287665B (en) | Ginkgolic acid-abamectin composite pesticide and preparation method thereof | |
| CN102659802B (en) | Application in terms of the preparation method of coumarin Neolignans and anti-marime fouling thereof | |
| CN101028322A (en) | Use of Maoliefengdou extract for preparing anti-cancer medicine | |
| CN101062062A (en) | Method for preparing anti-cancer effective component of marine animals | |
| CN104491048B (en) | A kind of loquat leaf total sesquiterpene glucoside extract and preparation method and application | |
| CN1955173A (en) | High purity cortex fraxini and its preparation method | |
| CN115286606A (en) | Gastrodia elata flavonoid compound and preparation method and application thereof | |
| CN112159440B (en) | Phenolic glycoside compound and preparation method and application thereof | |
| CN1864725A (en) | Method for extracting active ingredient of starofbethlehem and use thereof | |
| CN104262448B (en) | Method for extracting glycyrrhizic acid for licorice | |
| CN109929006B (en) | Extraction method and application of ergosterol peroxide in pleurotus ferulae | |
| CN105920065A (en) | Separation and purification method of begonia fimbritipula saponin and application of begonia fimbritipula saponin | |
| CN102295677A (en) | New nortriterpenoid saponin of Salicornia bigelovii Torr., preparation method and application thereof | |
| CN104664324A (en) | Preparation method and use of nucleoside effective part of sporocarp of artificial cultivation cordyceps militaris (L.) Link | |
| CN104523851A (en) | Method for extracting polygonum orientale pericarp general flavone extract product | |
| CN110680847A (en) | Method for extracting and purifying cannabinoids | |
| CN1597690A (en) | Enrichment method of rhodiola root glycoside compound | |
| CN1850816A (en) | Compound (2S)-5, 7.2', 6'-tetrahydroxy-6,8-diiso pentenyl-dihydro flavone extracting process and its use | |
| Formicki et al. | Effects of trigonella foenum graecum on the leukocytes in diabetes mice | |
| CN104398532B (en) | Application of cardiac glycoside compound 12beta-hydroxycalotropin | |
| CN102349937B (en) | Preparation method of ixeris sonchifolia hance extractive |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |