CN102772696A - Selenium cyclic peptide and fatty acid mixture and preparation method thereof - Google Patents
Selenium cyclic peptide and fatty acid mixture and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a selenium cyclic peptide and fatty acid mixture which is prepared from the traditional Chinese medicines of ginseng, radix rubiae, fructus lycii, fructus schisandrae chinensis, semen coicis, poria cocos, radix pseudostellariae, fructus gardeniae, selenium and the like by the modern microbial fermentation technology. The histone deacetylase all-natural synergic combined inhibitor with a brand-new chemical structure comprises the combined compounds such as selenium peptide, cyclic peptide, short-chain fatty acid and the like, and belongs to the tumor epigenetic regulation medicine of a novel anti-tumor action mechanism; and different from the traditional tumor treatment medicine, the selenium cyclic peptide and fatty acid mixture disclosed by the invention performs saturation attack by taking the abnormal epigenetic inheritance of tumor cells as a target spot, exerts an overall synergic anti-tumor effect by inducing the immunologic surveillance and immunity killing function of a patient, and is anti-oxidant and all-natural without chemical pollution.
Description
Technical field
The present invention relates to a kind of " mixture that contains selenium cyclic peptide, fatty acid " medicine and preparation method thereof, a kind of specifically selenium cyclic peptide mixture of fatty acid and preparation method thereof with anti-tumor oxidative function.
Background technology
Threatening the healthy great refractory disease of human life is tumor, and the mankind have experienced very long historical upheaval in the tumor treatment process.Antitumor drug is one of important treatment means, and along with going deep into and science and technology development of research, antitumor drug is constantly progressive along with the discovery of novel targets.
Scientist finds the further investigation of tumor, although tumor takes place and reasons of development is very complicated, all closely related unusually with gene expression and gene expression product activity after all.Simultaneously, because formed former the deterioration with the tumor surrounding (like angiogenesis, function of immune system variation abnormality etc.) of secondary of abnormal gene expression promotes pernicious apparent the further developing of tumor cell, and produces body internal diffusion and transfer.And the domestic and international research result clearly shows in recent years, and epigenetic changes in the generation of tumor and development and has general significance.Therefore, small-molecule drug research and development having become one of focus in international neoplasm targeted therapy field of carrying out to the important molecule target that influences epigenetic with this type of histon deacetylase (HDAC) (HDAC).
Because HDAC plays pivotal role in gene expression; So; They and many cell functions are closely related; Comprise cell cycle regulating, cell proliferation, immunoregulation, differentiation and gene program expression, have simultaneously with traditional chemotherapeutics to diverse adjusting pattern of drug resistance of tumor gene expression etc.Test shows that hdac inhibitor can suppress human body and the intravital growth of tumor of animal, comprises pulmonary carcinoma, gastric cancer, breast carcinoma, carcinoma of prostate and lymphoma etc.Hdac inhibitor also is different from typical targeted drug both being different from conventional cell poison type medicine (promptly to when breeding fast tumor cytotoxicity, also involving Normocellular traditional chemotherapeutics) on the mechanism of action.But new problem has appearred in present development work: (one), existing vegetalitas inhibitor content are low, weak effect; (2), the inhibitor performance of chemosynthesis is single, weak effect; (3), most of inhibitor is prone to degraded, half-life weak point, instability in vivo; (4), to the inhibition of transcribing of Hemopoietic factor GATA-1 and 10, can cause untoward reaction such as thrombocytopenia; (5), also have the trace element-selenium anti-tumor function certainly, but plasma selenium precipitates in the neutrality of small intestinal and slight alkali environment, has a strong impact on action effect.
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Summary of the invention
The objective of the invention is to provide a kind of antitumor, antioxidative vegetalitas selenium cyclic peptide mixture of fatty acid.
Another object of the present invention provides the method for preparing of this mixture.
The present invention is pathogenetic understanding and the Therapeutic Principle who " tumor " and concurrent oxidation resistance thereof is reduced according to Chinese medicine and pharmacy; Reach the pharmacological research achievement of " hdac inhibitor " with reference to modern " selenium ", " cyclic peptide ", " SCFA "; From the Chinese crude drug that can be used for health food of China's Ministry of Public Health authorization, filter out supplementing QI and nourishing YIN, blood circulation and channel invigorating, antitumor; Can improve the medical material of blood microcirculation and immunity again; Form raw material by Radix Ginseng, Radix Rubiae, Fructus Lycii, Fructus Schisandrae Chinensis, Semen Coicis, Poria, Radix Pseudostellariae, Fructus Gardeniae and trace element etc.; Extract with modern biotechnology enzyme process or microbe fermentation method: contain pure natural plant hdac inhibitor chemical compounds such as selenium peptide and cyclic peptide, SCFA, have efficient synergistic antitumor and antioxidation and the not synergetic effect of side effect.
Selenium cyclic peptide mixture of fatty acid is the subtype-selective albumen deacetylase pure natural composite restrainer with the brand-new chemical constitution that contains the selenium peptide; The tumor epigenetic regulation and control novel drugs that belongs to the new antitumoral mechanism of action; Different with traditional anti-tumor medicine; It carries out saturation attack with the unusual epigenetic of cancerous cell as target spot, and brings into play the overall coordination antitumor action through the immunologic surveillance and the immunologic cytotoxicity effect of inducing patient self, and pure natural does not have chemical contamination.
Article of the present invention are processed (consumption is a weight) by following component:
Radix Ginseng 2-15 part Radix Rubiae 3-20 part Fructus Lycii 5-25 part
Fructus Schisandrae Chinensis 2-10 part Semen Coicis 7-30 part Poria 5-20 part
Radix Pseudostellariae 8-30 part Fructus Gardeniae 4-20 part selenium 0.000001-0.002 part.
The optimum weight proportioning of article of the present invention is:
11 parts of 10 parts of Fructus Lycii of 5 portions of Radix Rubiaes of Radix Ginseng
8 parts in 15 parts of Poria of 4 parts of Semen Coiciss of Fructus Schisandrae Chinensis
0.0000029 part on 5 parts of selenium of 12 portions of Fructus Gardeniaes of Radix Pseudostellariae.
Can add other trace element in the above-mentioned prescription of article of the present invention,, generate the polypeptide metallo-chelate, improve functional effect as adding zinc, chromium etc.
The production method that above-mentioned each component is formed article of the present invention is: with above-mentioned magistral medicines material is raw material, adopts conventional pharmaceutical processing method to produce and contains selenium peptide, fatty acid Chinese patent medicine mixture, adopts the conventional method for preparing peptide to produce and contains the selenium oligopeptide mixture.Stress at present and select for use microbe fermentation method to prepare peptide, fatty acid cpds.Concrete grammar has two kinds: (one), above-mentioned prescription medical material mixed powder is broken, obtain decocting medicinal liquid (pretreatment) with the decocting method for distilling, add the peptide bacterial strain; Or comprehensive enzyme carries out the microbial fermentation cultivation, accent pH value, heating enzyme denaturing; Cooling; Filter purification, both, pharmaceutical dosage form processed by conventional method again.(2), (1) mix Radix Ginseng, Radix Rubiae, Fructus Schisandrae Chinensis, Poria, Radix Pseudostellariae and pulverize by above-mentioned formula proportion, with 75% ethanol (6 times of amounts) reflux, extract, 1.5 hours, the filtration alcohol extract was subsequent use, medicinal residues are preserved respectively; (2) Fructus Lycii, Semen Coicis, Fructus Gardeniae are mixed pulverizing by above-mentioned formula proportion, the medicinal residues after the adding alcohol extraction carry out decocting in 2 times (8 times of water), handle medical filtration in each 1.5 hours; (3) carry medicinal liquid to alcohol extraction medicinal liquid, water and mix back access microbial protease (millet straw bacillus); Under temperature 25-28 ° C, pH6.5-7, stirring condition, carried out enzymolysis 3-5 hour; Utilize existing little peptide in the plant that extracts; Under the effect of plant cyclase, optionally form cyclic peptide, enzyme denaturing; (4) above-mentioned medicinal liquid is concentrated, transfer ph value 7-10, stir at temperature 20-42 ° C, per minute under 60-80 time the condition and carry out anaerobic fermentation production SCFA, time 7-30 days; (5) cool off, filter purification, add the selenium powder suspension by above-mentioned formula proportion, the powerful stirring 2 hours, generation contains the selenium peptide, processes pharmaceutical dosage form by conventional method again, both gets selenium cyclic peptide mixture of fatty acid.
Antitumor principle of the present invention is through following mechanism: (one) is combined into the hdac inhibitor targeting by cyclic peptide, SCFA, the direct killing tumor cell; (2) contain selenium peptide Synergistic killing tumor cell; (3) promote immunity and oxidation resistance, suppress tumor cell existence; (4) block the blood supply of tumor area, stop tumor to continue to enlarge; (5) differentiation can be that a part of tumour cell transformation is a non-tumor cell.
Main feature of the present invention is: Caryophyllaceae, Rubiaceae, Schisandraceae, Araliaceae medical material contain cyclic peptide amount height in (one) prescription, and the cyclic peptide volume of production is many; (2) mixture contains binder (polysaccharide), pectin, fatty acid, amino acid derivativges, vegetable polysaccharides, polyhydric alcohol etc., is peptide absorption enhancer, dressing agent and stabilizing agent, so performance of the present invention is more stable, to human body safety, pharmacologically active is stronger; (3) bioactive peptide combines plasma selenium (containing the selenium peptide), discharges into blood again after being absorbed by small intestinal intestinal wall cell, thereby has avoided plasma selenium in the neutrality of small intestinal and slight alkali environment, to precipitate raising anti-tumor synergetic effect; (4) contained Chinese medicine and polypeptide have the effect of sticking between activated blood platelet.
In order to confirm the effect of the present invention to antitumor and antioxidation oxygen lack resistant function, we have done following test respectively.
(1) antitumor drug effect test
(1), medicament sources and preparation: selenium cyclic peptide mixture of fatty acid, 20 milliliters of oral liquids are pressed crude drug 1:1 and are extracted.With the preceding suspension that is mixed with the test desired concn with distilled water.
(2), experimental animal: body weight 18-22g kunming mice is adopted in test, male and female dual-purpose, but every batch of test sex is identical, is provided by Chinese Academy of Medical Sciences's animal center.
(3), test method: get well-grown mice transplantable tumor Lewis lung cancer, hepatocarcinoma H22 and sarcoma S180, press the conventional inoculation of anticarcinogen pharmacodynamic experiment specification requirement mice, in inoculation grouping administration in back 24 hours.Test is divided into five groups, and every group of 10 Mus are established matched group, cyclophosphamide positive drug group (conventional method), selenium cyclic peptide mixture of fatty acid 1.25,2.5,3.75g/kg three dose groups.Cyclophosphamide 60mg/kg lumbar injection once, continuous 9 times of selenium cyclic peptide mixture of fatty acid gastric infusion.Put to death animal in 24 hours after the drug withdrawal, the stripping tumor is weighed, and calculates tumour inhibiting rate, and carries out statistical procedures.
(4), result of the test:
1, to the clinical trial of Mice Bearing Lewis Lung Cancer:
Three batches of result of the tests, the tumour inhibiting rate of selenium cyclic peptide mixture of fatty acid 1.25g/kg group is respectively 31.0%, 26.3%, 25.2% (p<0.01, p<0.05, p<0.05); 2.5g/kg the tumour inhibiting rate of group is respectively 36.3%, 31.6%, 41.0% (p<0.01, p<0.01, p<0.001); 3.75g/kg the tumour inhibiting rate of group is respectively 51.0%, 38.3%, 41.2% (p<0.001, p<0.01, p<0.001).
2, selenium cyclic peptide mixture of fatty acid is to the clinical trial of rat liver cancer H22:
Three batches of result of the tests, the rate that presses down of 1.25g/kg group is respectively 16.7%, 36.1%, 23.9% (p<0.05, p<0.001, p<0.01); 2.5g/kg the tumour inhibiting rate of group is respectively 22.5%, 35.6%, 22.4% (p<0.05, p<0.001, p<0.05); 3.75g/kg the tumour inhibiting rate of group is respectively 29.6%, 41.4%, 34.2% (p<0.01, p<0.001, p<0.001).
3, selenium cyclic peptide mixture of fatty acid is to the clinical trial of murine sarcoma S180: three batches of result of the tests, the tumour inhibiting rate of selenium cyclic peptide mixture of fatty acid 1.25g/kg group are respectively 33.7%, 24.1%, 29.7% (p all < 0.05); 2.5g/>kg the tumour inhibiting rate of group is respectively 35.2%, 32.7%, 21.8% (p all < 0.05); 3.75g/>kg the tumour inhibiting rate of group be respectively 45.9%, 40.6%, 33.6% (p 0.05, p<0.01, p 0.05).
(5), conclusion (of pressure testing): selenium cyclic peptide mixture of fatty acid 1.25,2.5,3.75g/kg three dose groups have obvious tumor-inhibiting action to murine sarcoma S180, Mice Bearing Lewis Lung Cancer, hepatocarcinoma H22 have all been showed obvious tumor-inhibiting action, and be dose relationship.Wherein more obvious to the Mice Bearing Lewis Lung Cancer curative effect, the heavy dose of 4.74g/kg of selenium cyclic peptide mixture of fatty acid organizes average inhibitory rate 63.8%.
(2) antioxidation test
Selenium cyclic peptide mixture of fatty acid has the total flavones Chinese medicine ingredients, has anti-oxidation efficacy.We test to this.Method is following: with selenium cyclic peptide mixture of fatty acid (20 milliliters of oral liquids--1:1 crude drug extract), give to measure the biochemical indicator relevant with resisting oxidation and delaying senility behind aged BAL B/C mouse stomach 45 d.The result: selenium cyclic peptide mixture of fatty acid can make liver superoxide dismutase (SOD) activity significantly strengthen; Whole blood glutathion peroxidase (GSH-Px) and catalase (CAT) activity significantly improve.Conclusion: selenium cyclic peptide mixture of fatty acid has significantly antioxidation.
Embodiment: take by weighing raw material by following prescription ratio
11 parts of 10 parts of Fructus Lycii of 5 portions of Radix Rubiaes of Radix Ginseng
8 parts in 15 parts of Poria of 4 parts of Semen Coiciss of Fructus Schisandrae Chinensis
0.0000029 part on 5 parts of selenium of 12 portions of Fructus Gardeniaes of Radix Pseudostellariae.
The embodiment production method:
(1) Radix Ginseng, Radix Rubiae, Fructus Schisandrae Chinensis, Poria, Radix Pseudostellariae are mixed by above-mentioned formula proportion pulverize, with 75% ethanol (6 times of amounts) reflux, extract, 1.5 hours, the filtration alcohol extract was subsequent use, and medicinal residues are preserved respectively; (2) Fructus Lycii, Semen Coicis, Fructus Gardeniae are mixed pulverizing by above-mentioned formula proportion, the medicinal residues after the adding alcohol extraction carry out decocting in 2 times (8 times of water), handle medical filtration in each 1.5 hours; (3) carry medicinal liquid to alcohol extraction medicinal liquid, water and mix back access microbial protease (millet straw bacillus), under temperature 25-28 ° C, pH6.5-7, stirring 200rpm condition, carried out enzymolysis 4 hours, produce cyclic peptide, enzyme denaturing; (4) above-mentioned medicinal liquid is concentrated, transfer ph value 7-10, under the condition of 40 ° of C of temperature, per minute stirring 80 times, carry out anaerobic fermentation and acid production (SCFA), 10 days time; (5) filter purification; Medicinal liquid is concentrated to takes 3 times, each 20 milliliters concentration (medical material: water is 1:0.86) for each person every day, add the selenium powder suspension by above-mentioned formula proportion again, the powerful stirring 2 hours; Generation contains the selenium peptide; Envelope is pricked in fill in time, 105 ° of C flowing steam sterilizations 30 minutes, both selenium cyclic peptide mixture of fatty acid.
Claims (3)
1. antitumor antioxidative selenium cyclic peptide mixture of fatty acid is characterized in that it is the mixture of being processed by following weight proportion raw material and method:
Radix Ginseng 2-15 part Radix Rubiae 3-20 part Fructus Lycii 5-25 part
Fructus Schisandrae Chinensis 2-10 part Semen Coicis 7-30 part Poria 5-20 part
Radix Pseudostellariae 8-30 part Fructus Gardeniae 4-20 part selenium 0.000001-0.002 part
Process coarse powder with pulverizing after the raw materials mix of said ratio, obtain the decocting medicinal liquid, add enzyme and carry out the microbial fermentation cultivation, transfer pH value with conventional method for distilling, the heating enzyme denaturing, purification is filtered in cooling, both gets, and processes pharmaceutical dosage form by conventional method again.
2. mixture according to claim 1; It is characterized in that said method for preparing may further comprise the steps: (1) is mixed pulverizing with Radix Ginseng, Radix Rubiae, Fructus Schisandrae Chinensis, Poria, Radix Pseudostellariae by above-mentioned formula proportion; With 75% ethanol (6 times of amounts) reflux, extract, 1.5 hours; The filtration alcohol extract is subsequent use, and medicinal residues are preserved respectively; (2) Fructus Lycii, Semen Coicis, Fructus Gardeniae are mixed pulverizing by above-mentioned formula proportion, the medicinal residues after the adding alcohol extraction carry out decocting in 2 times (8 times of water), handle medical filtration in each 1.5 hours; (3) carry medicinal liquid to alcohol extraction medicinal liquid, water and mix back access microbial protease, under temperature 25-28 ° C, pH6.5-7, stirring condition, carried out enzymolysis 3-5 hour, produce peptide, enzyme denaturing; (4) above-mentioned medicinal liquid is concentrated, transfer ph value 7-10, stir at temperature 20-42 ° C, per minute under 60-80 time the condition and carry out anaerobic fermentation production SCFA, time 7-30 days; (5) purification is filtered in cooling, adds selenium by above-mentioned formula proportion, processes pharmaceutical dosage form by conventional method again, has both got selenium cyclic peptide mixture of fatty acid.
3. mixture according to claim 1 and 2 is characterized in that said mixture can be processed into health food.
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Cited By (2)
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CN106957848A (en) * | 2016-01-08 | 2017-07-18 | 贵阳中医学院 | One Radix pseudostellariae cyclic peptides HB precursor protein gene and its application |
CN113768833A (en) * | 2021-08-16 | 2021-12-10 | 楚香(上海)生物科技有限公司 | Pure natural deacetylase activator and application thereof |
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CN1344806A (en) * | 2001-09-29 | 2002-04-17 | 邹远东 | Enzyme process of preparing small molecule active coix seed polypeptide |
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Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN106957848A (en) * | 2016-01-08 | 2017-07-18 | 贵阳中医学院 | One Radix pseudostellariae cyclic peptides HB precursor protein gene and its application |
CN106957848B (en) * | 2016-01-08 | 2021-12-17 | 贵州中医药大学 | Heterophylly falsestarwort root cyclic peptide HB precursor protein gene and application thereof |
CN113768833A (en) * | 2021-08-16 | 2021-12-10 | 楚香(上海)生物科技有限公司 | Pure natural deacetylase activator and application thereof |
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