CN1506067A - Application of chitin oligose in anticancer drug - Google Patents
Application of chitin oligose in anticancer drug Download PDFInfo
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- CN1506067A CN1506067A CNA021447063A CN02144706A CN1506067A CN 1506067 A CN1506067 A CN 1506067A CN A021447063 A CNA021447063 A CN A021447063A CN 02144706 A CN02144706 A CN 02144706A CN 1506067 A CN1506067 A CN 1506067A
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Abstract
The present invention relates to anticancer drug, and is especially the application of chitin oligose in anticancer drug. The chitin oligose has a polymerization degree of 2-50, component of polymerization degree of 4-30 not less than 50 %, and purity higher than 98 %. The contrast research of the cancer suppressing rate of the chitin oligose on liver cancer, large intestine cancer, cervical cancer, erythroleukemia, lymph leukemia and other tumor cells shows that the chitin oligose has strongest cancer suppressing rate, 79 %, on liver cancer. The primary observation shows that the cancer suppressing effect of chitin oligose is related with its inducing cell death. Medicine level chitin oligose and chitin oligose medicine have the functions of suppressing cancer cell, suppressing cancer cell metastasis and suppressing cancer toxin.
Description
Technical field
The present invention relates to the preparation and the application of cancer therapy drug, the application of oligochitosan in cancer therapy drug specifically.
Background technology
Cancer is one of important diseases of harm people's life health because its course of disease is short, harm is big, popular wide.Still lack ideal medicine at present; According to the investigation of 29 provinces, cities and autonomous regions' populations more than 800,000,000, China has approximately greater than the hundreds of thousands people die from nine big common cancers every year; Mainly contain surgical operation, chemotherapy, radiotherapy, gene therapy, Chinese medicine and interventional therapy for treatment for cancer; But because cancer often merges severe complication or sends out in vivo, the excision rate only is about 5%, has lost the surgical engine meeting when nearly 95% patient makes a definite diagnosis, relies on Drug therapy mostly; So the medicine of developmental research treatment cancer has the meaning of particular importance; Chinese Academy of Sciences's Dalian chemistry institute proposes an application for a patent for invention on January 5th, 2000, application number is the be coupled method of production oligochitosan of 00110009.2 1 kinds of enzymic degradation chitosans and membrance separation; This invention combines membrane separation technique with the enzymic degradation chitosan, promptly adopt doughnut or flat membrane ultrafiltration device that degradation reaction liquid original position is separated, in time isolate oligochitosan with physiologically active, control the further degraded of active shell oligosaccharide under the enzyme effect effectively, also can prevent the inhibitory action of catabolite to reaction, because realize continued operation, enzyme is reusable, improve the service efficiency of digestive enzyme.The filter liquor of ultrafilter membrane is again through nanofiltration, removes the oligosaccharide of non-activity of low polymerization degree and a large amount of water, and ultrafiltrate is concentrated, and obtains the oligochitosan product of certain high degree of polymerization scope of physiologically active.Nowadays, development has the research and development of oligochitosan (the being amino-oligosacchride) PTS of China's independent intellectual property right; It will be significant to the improvement of human health, economy and social development and environment.
Summary of the invention
The object of the present invention is to provide the application of oligochitosan in cancer therapy drug, its hepatocarcinoma suppression ratio is higher.
For achieving the above object, the technical solution used in the present invention is: utilizing application number is 00110009.2 reaction separation coupling technology, and the nanometer film technology is applied in concentrating of oligochitosan and the purification; By ethanol precipitation, the preparation degree of polymerization is 2～50 pharmaceutical grade oligochitosans then, and wherein the degree of polymerization is that 4～30 content are not less than 50%, does not comprise purity＞98% of the oligochitosan of moisture, is used for cancer therapy drug;
Can be made into multiple dosage forms such as water preparation, powder, tablet and drop pill.
Wherein said oligochitosan preparation process is as follows: at first chitosan is dissolved in 1～10% the acetum, add substrate and weigh 1～10% chitosanase, intensification is stirred to 20～60 ℃ of constant temperature, begin ultrafiltration after 10～60 hours, part less than molecular cut off (300) sees through film, molecule greater than molecular cut off (6000) returns reactor continuation degraded, the ultrafiltration permeate is again by the nanofiltration device, permeation parts is a water, acetic acid and small molecular sugar, to be condensed to not be that 10～50% degree of polymerization are 2～50 oligochitosans to permeation parts, through 70～100% ethanol precipitation, filters then, supernatant forms 2～50 medicine level oligochitosans through spray drying.
The present invention has following advantage:
1. the oligochitosan of the present invention's employing is to the various human tumor cell line (finding through a large amount of experiments) of In vitro culture, show as the tumour inhibiting rate comparative study of oligochitosan to tumor cells such as people's hepatocarcinoma, colorectal cancer, cervical cancer, erythroleukemia, leukemic lymphoblastoids: its inhibitory action to hepatocarcinoma is the strongest, and tumour inhibiting rate is 79% (0.1mg/ml).Preliminary observation may be relevant with inducing apoptosis of tumour cell to the oligochitosan antitumaous effect.
2. pharmaceutical grade oligochitosan of the present invention and oligochitosan medicine have anticancer, anticancer transfer, suppress the effect of carcinomycin.
3. the present invention is under the support of the Ministry of Science and Technology " 95 " brainstorm project, adopts enzyme reaction separation coupling technology and alcohol precipitation method to prepare the new technology that the degree of polymerization is 2～50 usefulness oligochitosans.
Description of drawings
Fig. 1. the process chart of medicinal oligochitosan preparation.
The specific embodiment
Below by example technology of the present invention is further specified:
Embodiment 1 medicinal oligochitosan preparation
At first 5% chitosan is dissolved in 4% acetum, add substrate (with respect to chitosan) and weigh 5% chitosanase, intensification is stirred to 30 ℃ of reactions down, begin ultrafiltration after 60 hours, part less than molecular cut off (300) sees through film, molecule greater than molecular cut off (6000) returns reactor continuation degraded, the ultrafiltration permeate is again by the nanofiltration device, permeation parts is a water, acetic acid and small molecular sugar, to be condensed to not be that 50% degree of polymerization is 2～50 oligochitosans to permeation parts, through 75% ethanol precipitation, filters then, supernatant forms 2～50 medicine level oligochitosans through spray drying; Wherein the degree of polymerization is 4～30 content 65%, does not comprise the purity 98.5% of the oligochitosan of moisture.Its technological process as shown in Figure 1.
The acquisition product is applied to following anticancer therapeutic test
Embodiment 2 oligomeric chitosans are to the synthetic inhibitory action of cancerous cell DNA
The oligomeric chitosan of table 1. is to the synthetic inhibitory action of cancerous cell DNA
| Sample | Concentration | Suppression ratio (%) |
| Oligochitosan | Stock solution (12g/100ml) | ????87.5 |
| Dilute 10 times | ????81.3 | |
| Dilute 100 times | ????79.7 | |
| Contrast | ????13.3 | |
| ????13.5 | ||
| ????43.7 |
As can be seen from Table 1, oligochitosan to the comparison of the synthetic inhibitory action of cancerous cell DNA according to (clear water) high many.
Embodiment 3 oligochitosans are to the kinds of tumor cells extracorporeal anti-tumor function
Table 2. oligochitosan is to the kinds of tumor cells extracorporeal anti-tumor function
| The oligochitosan group | The cisplatin group | The normal control group | |||||
| ????1mg/ml | ????400 | ????100 | ???10 | ????10(μg/ml) | (not dosing) | ||
| Hepatocarcinoma | ??OD | ????0.33 | ????0.27 | ????0.24 | ????0.49 | ????0.28 | ????1.12 |
| ??％ | ????73 | ????76 | ????79 | ????56 | ????75 | ||
| Hela | ??OD | ????0.49 | ????0.51 | ????0.65 | ????0.73 | ????0.15 | ????0.96 |
| ??％ | ????49 | ????47 | ????32 | ????24 | ????84 | ||
| K562 | ??OD | ????0.48 | ????0.51 | ????117 | ????1.31 | ????0.65 | ????1.50 |
| ??％ | ????68 | ????66 | ????22 | ????13 | ????57 | ||
| Raji | ??OD | ????024 | ????0.54 | ????0.76 | ????0.69 | ????0.15 | ????0.81 |
| ??％ | ????70 | ????33 | ????6 | ????15 | ????82 | ||
| Hepa | ??OD | ????0.33 | ????0.48 | — | — | ????0.05 | ????0.98 |
| ??％ | ????66 | ????51 | ????95 |
Cervical cancer (Hela), leukemia (K562), people cling to leukemia (Raji), rat liver cancer ascites (Hepa)
As can be seen from Table 2, oligochitosan has obvious inhibitory action to hepatoma carcinoma cell, simultaneously leukemia and Hepar Mus cancer ascites cells is also had certain inhibitory action.
The experiment of embodiment 4 oligochitosan vitro inhibition mice HCa-F tumors
The experiment of table 3. oligochitosan vitro inhibition mice HCa-F tumor
| Oligochitosan | ?10μg/ml | ?50μg/ml | ?100μg/ml | ?400μg/ml | Cisplatin 10 μ g/ml | 1% acetic acid |
| 3 | ?5±0 | ?32±3.53 | ?77.5±10.6 | ?100±0 | ?20±0 | ?5±0 |
| 6 | ?7.5±3.53 | ?45±7.07 | ?100±0 | ?100±0 | ?32.5±3.53 | ?5±0 |
| 24 | ?12.5±6.45 | ?65±7.07 | ?100±0 | ?100±0 | ?95±0 | ?10±0 |
As can be seen from Table 3, oligochitosan has stronger killing in vitro ability to high lymphatic metastasis cancerous cell line Hca-F, and the tumor ability that presses down of 100 μ g/ml oligochitosans is better than 10 μ g/ml cisplatin, and the tumor ability that presses down of 50 μ g/ml oligochitosans is weaker than 10 μ g/ml cisplatin slightly.The solvent acetic acid of oligochitosan does not have the tumor ability of pressing down.
Embodiment 5 oligochitosans to the inhibition of S-180 and with the mannooligo saccharide specific activity
Table 4. oligochitosan to the inhibition of S-180 and with the mannooligo saccharide specific activity
| Oligosaccharide | Concentration | Suppression ratio (%) |
| Mannooligo saccharide | ????5mg/ml | ????59.5 |
| ????3mg/ml | ????60.5 | |
| ????2.5mg/ml | ????69.4 | |
| ????1.5mg/ml | ????12.9 | |
| The sulfonation mannooligo saccharide | ????5mg/ml | ????18.8 |
| ????2.5mg/ml | ????32.6 | |
| ????1.5mg/ml | ????34.8 | |
| Oligochitosan | ????9.5mg/ml | ????90.5 |
| ????7.14mg/ml | ????98.9 | |
| ????4.76mg/ml | ????99.5 |
Mouse ascites cancer (S-180)
As can be seen from Table 4, three kinds of oligosaccharide comparative test results show: the effect of oligochitosan inhibition S-180 is best, secondly is mannooligo saccharide and sulfonation mannooligo saccharide.
Embodiment 6 oligochitosans are to the liver cancer cell growth inhibitory action
Table 5. oligochitosan is to the liver cancer cell growth inhibitory action
| Handle | Concentration (μ g/m1) | A595nm light absorption value (X) | |
| Non-oxide shell oligosaccharide solution | Oxidized form oligochitosan solution | ||
| The oligochitosan group | ?????30.0 | ??????0.553 | ??????0.274 |
| ?????18.0 | ??????1.032 | ??????0.269 | |
| ?????10.8 | ??????1.327 | ??????0.317 | |
| ?????6.5 | ??????1.415 | ??????0.382 | |
| The cisplatin group | ?????10.0 | ??????0.083 | ??????0.080 |
| Matched group | ??????1.621 | ??????1.468 | |
As can be seen from Table 5, this experiment confirms that further oligochitosan has obvious inhibitory action to hepatoma carcinoma cell, tests used non-oxide type and compares with matched group with oxidized form oligochitosan solution, all hepatoma carcinoma cell is had obvious inhibitory action, and be dose-effect relationship, especially more obvious with the effect of oxidized form oligochitosan.
The influence that embodiment 7 oligochitosans are heavy to the solid tumor mouse tumor
The influence that table 6. oligochitosan is heavy to the solid tumor mouse tumor
| Group | Dosage (mL/kg.bw) | Number of animals (only) | Solid tumor heavy (g) | The P value |
| The blank group | 0 | ?16 | ?1.8807±0.7513 | — |
| Low dose group | 3.3 | ?16 | ?1.4212±1.1770 | 0.2500 |
| Middle dosage group | 6.7 | ?14 | ?1.2836±1.1200 | 0.1232 |
| High dose group | 20.0 | ?16 | ?0.6110±0.3570 *** | 2.0*10 -6 |
| Positive controls | 0 | ?15 | ?1.2071±0.7301 * | 0.0263 |
*: with matched group significant difference (p＜0.05) μ is arranged relatively
* *: with matched group utmost point significant difference (p＜0.001) is arranged relatively
As can be seen from Table 6, per os gave the oligomeric chitosan of mice various dose after 30 days, compare with matched group, and the lotus ascites tumor mice prolonged survival period 34% of middle dosage group (6.7mL/kg.bw), repeated experiments prolongs 37%; Lotus solid tumor mouse tumor heavily reduces by 63%, and repeated experiments reduces by 30%.The lotus ascites tumor mice prolonged survival period 35% of high dose group (20.0mL/kg.bw), repeated experiments prolongs 36%; Lotus solid tumor mouse tumor heavily reduces by 76%, and repeated experiments reduces by 67%.
The influence of 8 pairs of normal macrophage phagocytosis of mice of embodiment
The influence of table 7. pair normal macrophage phagocytosis of mice
| Group | Dosage (mL/kg.bw) | Number of animals (only) | ???????a | The P value |
| The blank group | ??????0 | ????12 | ??6.55±0.29 | ??— |
| Low dose group | ??????3.3 | ????12 | ??6.51?±0.45 | ??0.7631 |
| Middle dosage group | ??????6.7 | ????12 | ??6.40±0.69 | ??0.4805 |
| High dose group | ?????20.0 | ????12 | ??6.56±0.47 | ??0.9884 |
By table 7 as seen, what per os gave the mice various dose was tried thing after 30 days, with matched group relatively, the carbon of each dosage group is cleaned up index does not all have significant difference (p＞0.05).
Embodiment 9 oligomeric chitosans are to the influence of the active NK cell of mice
The oligomeric chitosan of table 8. is to the influence of the active NK cell of mice
| Group | Dosage (mL/kg.bw) | Number of animals (only) | NK activity (1: 50) (%) |
| The blank group | ??????0 | ????12 | ????32.8±10.6 |
| Low dose group | ??????3.3 | ????12 | ????29.6±12.7 |
| Middle dosage group | ??????6.7 | ????12 | ????28.0±12.0 |
| High dose group | ??????20.0 | ????12 | ????33.4±10.9 |
Lymphocyte (NK cell)
By table 7 as seen, middle dosage group and high dose group and matched group comparison carbon are cleaned up index, the NK cytoactive does not all have significant difference (p＞0.05).Experimental result shows oral middle dosage or the effect of high dose oligochitosan, can not cause damage to body immune system during taking.
Claims (3)
1. the application of oligochitosan in cancer therapy drug is characterized in that: the degree of polymerization of oligochitosan is 2～50, and wherein the degree of polymerization is that 4～30 content are not less than 50%, does not comprise purity＞98% of the oligochitosan of moisture.
2. according to the application of the described oligochitosan of claim 1 in cancer therapy drug, it is characterized in that: wherein said oligochitosan preparation process is as follows: at first chitosan is dissolved in 1～10% the acetum, add substrate and weigh 1～10% chitosanase, intensification is stirred to 20～60 ℃ of constant temperature, begin ultrafiltration after 10～60 hours, the aperture of film is 300～6000, part less than molecular cut off sees through film, molecule greater than molecular cut off returns reactor continuation degraded, the ultrafiltration permeate is again by the nanofiltration device, permeation parts is a water, acetic acid and small molecular sugar, to be condensed to not be that 10～50% degree of polymerization are 2～50 oligochitosans to permeation parts, through 70～100% ethanol precipitation, filters then, supernatant forms 2～50 medicine level oligochitosans through spray drying.
3. according to the application of the described oligochitosan of claim 1 in cancer therapy drug, it is characterized in that: wherein said oligochitosan cancer therapy drug can be made into water preparation, powder, tablet and the drop pill that suppresses hepatocarcinoma, colorectal cancer, cervical cancer, erythroleukemia, leukemic lymphoblastoid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA021447063A CN1506067A (en) | 2002-12-06 | 2002-12-06 | Application of chitin oligose in anticancer drug |
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| Application Number | Priority Date | Filing Date | Title |
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| CNA021447063A CN1506067A (en) | 2002-12-06 | 2002-12-06 | Application of chitin oligose in anticancer drug |
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| CN1506067A true CN1506067A (en) | 2004-06-23 |
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| CNA021447063A Pending CN1506067A (en) | 2002-12-06 | 2002-12-06 | Application of chitin oligose in anticancer drug |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1300158C (en) * | 2005-03-22 | 2007-02-14 | 重庆邮电学院 | Non-natural active oligosaccharide compounds, preparing process and application |
| CN102225198A (en) * | 2011-06-28 | 2011-10-26 | 南京农业大学 | Anti-tumour medicine composition and preparation method and application thereof |
| WO2017107627A1 (en) * | 2015-12-24 | 2017-06-29 | 广东药科大学 | Chitosan oligosaccharide tablet and preparation method therefor |
| CN107158024A (en) * | 2017-06-21 | 2017-09-15 | 哈尔滨工业大学 | A kind of application of chitosan oligosaccharide |
| CN107373674A (en) * | 2017-08-25 | 2017-11-24 | 西宝生物科技（上海）股份有限公司 | Alimentation composition and its application after a kind of oncotherapy |
| CN108553425A (en) * | 2018-06-01 | 2018-09-21 | 广东药科大学 | A kind of chitosan oligosaccharide dripping pill and preparation method thereof |
| CN110279862A (en) * | 2019-07-09 | 2019-09-27 | 上海市第六人民医院 | A kind of anti-cancer composition and its application in the drug of preparation treatment osteosarcoma |
| CN110742897A (en) * | 2019-12-04 | 2020-02-04 | 山东陆海蓝圣生物科技股份有限公司 | Chitosan oligosaccharide and curcumin compound and application thereof |
-
2002
- 2002-12-06 CN CNA021447063A patent/CN1506067A/en active Pending
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1300158C (en) * | 2005-03-22 | 2007-02-14 | 重庆邮电学院 | Non-natural active oligosaccharide compounds, preparing process and application |
| CN102225198A (en) * | 2011-06-28 | 2011-10-26 | 南京农业大学 | Anti-tumour medicine composition and preparation method and application thereof |
| CN102225198B (en) * | 2011-06-28 | 2013-01-23 | 南京农业大学 | Anti-tumour medicine composition and preparation method and application thereof |
| WO2017107627A1 (en) * | 2015-12-24 | 2017-06-29 | 广东药科大学 | Chitosan oligosaccharide tablet and preparation method therefor |
| CN107158024A (en) * | 2017-06-21 | 2017-09-15 | 哈尔滨工业大学 | A kind of application of chitosan oligosaccharide |
| CN107373674A (en) * | 2017-08-25 | 2017-11-24 | 西宝生物科技（上海）股份有限公司 | Alimentation composition and its application after a kind of oncotherapy |
| CN108553425A (en) * | 2018-06-01 | 2018-09-21 | 广东药科大学 | A kind of chitosan oligosaccharide dripping pill and preparation method thereof |
| WO2019227743A1 (en) * | 2018-06-01 | 2019-12-05 | 广东药科大学 | Chitosan oligosaccharide dropping pill and preparation method therefor |
| CN110279862A (en) * | 2019-07-09 | 2019-09-27 | 上海市第六人民医院 | A kind of anti-cancer composition and its application in the drug of preparation treatment osteosarcoma |
| CN110279862B (en) * | 2019-07-09 | 2022-04-08 | 上海市第六人民医院 | Anticancer composition and application thereof in preparing medicine for treating osteosarcoma |
| CN110742897A (en) * | 2019-12-04 | 2020-02-04 | 山东陆海蓝圣生物科技股份有限公司 | Chitosan oligosaccharide and curcumin compound and application thereof |
| CN110742897B (en) * | 2019-12-04 | 2023-07-14 | 山东陆海蓝圣生物科技股份有限公司 | Compound of chitosan oligosaccharide and curcumin and application thereof |
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