CN103212087A - Antibiotic-chitosan covalent conjugate and preparation method and application thereof in preparation of anti-tumor medicines - Google Patents
Antibiotic-chitosan covalent conjugate and preparation method and application thereof in preparation of anti-tumor medicines Download PDFInfo
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Abstract
The invention discloses an antibiotic-chitosan covalent conjugate and a preparation method and application thereof in preparation of anti-tumor medicines. The preparation method comprises the following steps of: (1) preparing a chitosan water solution and an antibiotic water solution, mixing in a mole ratio of chitosan to antibiotic being 1:0.5-1:10, adding NaCNBH3, stirring by shielding sunlight, and reacting for 15-72 hours at 15-30 DEG C; (2) transferring the water solution obtained by the step (1) to a dialysis bag with molecular cut off of 1400-10000, and performing dialysis for 1-5 days to obtain a dialysis liquid; and (3) drying the dialysis liquid obtained by the step (2). The obtained antibiotic-chitosan covalent conjugate has stronger damaging effect to tumor cells, treats tumors in wide range, and is exact in effective components.
Description
Technical field
The present invention relates to medical technical field, in particular a kind of antibiotic-chitosan covalent complex and preparation method thereof and the application in the preparation antitumor drug.
Background technology
Human life in the malignant tumor serious threat.According to national resident's coroner's inquest result demonstration for the third time, the mortality rate that China's tumor causes occupies second in all causes of disease, account for 17.9%, and sickness rate is in rising trend.Over nearly 20 years, China's tumor mortality rate has risen 29.42%.In 35～59 years old middle prime of life crowd, tumor has been listed as and has occupied first of all kinds of causes of the death.At present in the whole world and even China, cancer has become and has caused human dead second largest reason.
In recent years, chemotherapy of tumors had been obtained suitable progress, and the tumor patient life span obviously prolongs, and the chemical classes antineoplastic agent plays an important role therein.The chemical classes antitumor drug is meant the medicine that acts on the DNA chemical constitution, belongs to cytotoxic drug, comprises the antibiotic of alkylating agent, metal platinum complex, anthracene ring antitumor medicinal and destruction DNA etc.Common antitumor antibiotics has polypeptide antibiotics (as actinomycin D and bleomycin) and anthraquinone class antibiotic (as amycin and mitoxantrone hydrochloride), yet above these traditional tumour medicines all exist poor specificity, side effect is obvious, the shortcoming that price is high.
Chitosan (chitosan) is the chemical compound that is obtained through deacetylation by the chitin that nature extensively exists, in medicine, food, chemical industry, cosmetics, water treatment, METAL EXTRACTION and recovery, biochemistry and biomedical engineering extensive use, streptomycin (streptomycin) be behind penicillin second produce and be used for clinical a kind of glucosamine type antibiotic, cheap and easy to get.
Summary of the invention
Technical problem to be solved by this invention is all to exist poor specificity at present traditional clinically tumour medicine, side effect is obvious, the shortcoming that price is high provides a kind of antibiotic-chitosan covalent complex and preparation method thereof and the application in the preparation antitumor drug.
For solving the problems of the technologies described above, one of technical scheme that the present invention takes is: the preparation method of a kind of chitosan-antibiotic covalent complex, and described preparation method may further comprise the steps:
(1) preparation chitosan and antibiotic aqueous solution are mixing in 1: 0.5～1: 10 with chitosan and antibiotic according to molar ratio, add NaCNBH
3, lucifuge stirs, and 15 ℃～30 ℃ were reacted 15～72 hours;
(2) aqueous solution of step (1) gained being moved into molecular cut off is in 1400～10000 the bag filter, to dialyse 1～5 day, obtains dialysis solution;
(3) with the dialysis solution drying of step (2) gained, promptly.
Wherein the described chitosan of step (1) is the conventional chitosan in this area.Described chitosan (chitosan).The preparation method of described chitosan preferably is: the chitin (chitin) that is extensively existed by nature prepares through deacetylation.The chemical name of described chitosan is polydextrose amine (1-4)-2-amino-B-D glucose.Wherein said antibiotic is the antibiotic of this area routine.Described antibiotic preferably is an aminoglycoside antibiotics.Described aminoglycoside antibiotics is the glucosides class antibiotic that is formed by connecting by oxo bridge by amino sugar and aminocyclitol.Described antibiotic preferably is streptomycin, kanamycin or gentamycin.Antibiotic of the present invention is preferably streptomycin.
Described chitosan of step (1) and antibiotic aqueous solution are formed mixed liquor, and described chitosan and antibiotic mol ratio more preferably are 1: 1～1: 6, are 1: 6 best.The time of described reaction more preferably is 10～48 hours, is 24 hours best.The temperature of described reaction more preferably is 15 ℃～30 ℃, is 20 ℃ best.
The described NaCNBH of step (1) wherein
3Be gentle Reducing agent, be used for reducing carbon-to-nitrogen double bon between streptomycin and chitosan, reaction forward is carried out, can not replace with other Reducing agent, addition preferably is 30%～50% of a described antibiotic content, and addition is 30% best, and described percentage ratio is mass percent.
Wherein the described dialysis of step (2) is this area conventional dialysis method.Described dialysis is a kind of selectivity diffusion process of passing film, can be used for the solute that the isolated molecule amount varies in size, be lower than film and hold back the material of threshold value molecular weight and can diffuse through film, be higher than film and hold back the opposite side that the material of threshold value molecular weight then is retained in semipermeable membrane.Wherein said bag filter is this area conventional dialysis bag.The molecular cut off of wherein said bag filter preferably is 3000～5000, more preferably is 3500.The time of wherein said dialysis more preferably is 2～3 days, and dialysis time is 2 days best.
Wherein the described drying of step (3) is this area conventional drying mode.Described drying preferably is vacuum lyophilization, vacuum drying, spray drying, oven dry or infrared drying, is vacuum lyophilization best.The parameter of described vacuum lyophilization preferably is: temperature-35 ℃～-45 ℃, vacuum 20～30Pa, 36～48 hours time.
For solving the problems of the technologies described above, two of the technical scheme that the present invention takes is: aforesaid preparation method prepares the chitosan-antibiotic covalent complex of gained.
Chitosan and antibiotic mol ratio preferably are 1: 1～1: 10 in described chitosan-antibiotic covalent complex, more preferably are 1: 1～1: 6, are 1: 6 best.
For solving the problems of the technologies described above, three of the technical scheme that the present invention takes is: the purposes of aforesaid chitosan-antibiotic covalent complex in the preparation antitumor drug.
Wherein said tumor is the conventional tumor in this area.Described tumor preferably comprises: leukemia, hepatocarcinoma, pulmonary carcinoma, skin carcinoma, gastric cancer, hepatocarcinoma, oral cancer or lymphoma.
Purposes of the present invention preferably is meant chitosan-antibiotic covalent complex as active component, makes the antitumor drug of various dosage forms with acceptable accessories.Wherein said acceptable accessories is the conventional adjuvant that uses in this area.Described excipient substance is meant excipient and additives used when producing medicine and prescription being dispensed.
On the basis that meets this area general knowledge, above-mentioned each optimum condition, but combination in any promptly get the preferred embodiments of the invention.
Agents useful for same of the present invention and raw material are all commercially available to be got.
Positive progressive effect of the present invention is: the raw material for preparing chitosan of the present invention-antibiotic covalent complex is easy to get, and simple synthetic method is easily gone; Prove that through cell experiment gained chitosan-antibiotic covalent complex has stronger destruction to tumor cell, wide at the kind scope of tumor, effective ingredient is clear and definite; Described chitosan-antibiotic covalent complex can effectively improve tumor to the antibiotic sensitivity of tradition, reduces traditional antibiotic use amount, and has expanded traditional antibiotic range of application.
Description of drawings
Fig. 1 is chitosan-streptomycin covalent complex molecular weight determination collection of illustrative plates.
Fig. 2 is chitosan-streptomycin covalent complex infared spectrum.
Fig. 3 is the nuclear magnetic resonance, NMR C spectrum of chitosan-streptomycin covalent complex.
Fig. 4 is the nuclear magnetic resonance, NMR H spectrum of chitosan-streptomycin covalent complex.
Fig. 5 is the inhibitory action design sketch of chitosan-streptomycin covalent complex to growth of tumour cell, wherein A and B distinguish representative stomach cancer cell SGC7901 and BGC823, C representative colon cancer HT29, D representative colorectal cancer Lovo cell, E, F and G be representative's hepatoma carcinoma cell BEL-7402, SMMC-7721 and HepG2 respectively, H representative lung cell A549, I representative esophageal cancer cell Eca-109, J represents the Hela cell, K representative dermal melanin oncocyte A375, L representative osteosarcoma cell MG63.
Fig. 6 is chitosan-streptomycin covalent complex to the inhibitory action design sketch of gastric carcinoma cells SGC7901 growth, and wherein to represent drug treating time be 6h to A, and the B role of delegate time is 12h, and the C role of delegate time is 24h.
Fig. 7 is the inhibitory action of different proportion chitosan-streptomycin covalent complex to human lung cancer cell A549's growth, wherein A represented chitosan-streptomycin covalent complex 1: 1, B represented chitosan-streptomycin covalent complex 1: 2, C represented chitosan-streptomycin covalent complex 1: 4, and D represented chitosan-streptomycin covalent complex 1: 6.
Fig. 8 is the inhibitory action that different chitosan-antibiotic covalent complexes is grown to the human lung cancer cell A549, wherein A represents chitosan-acetylspiramycin covalent complex, B represents chitosan-clarithromycin covalent complex, and C represents chitosan-erythromycin covalent complex.
The specific embodiment
Below in conjunction with specific embodiment, the present invention is described in detail.Agents useful for same of the present invention and raw material are all commercially available to be got.The experimental technique of unreceipted actual conditions in the following example according to conventional method and condition, or is selected according to catalogue.
Embodiment 1
Get chitosan 100mg, streptomycin 906mg is dissolved in respectively in the 30ml deionized water, and adds 0.744g sodium cyanoborohydride (NaCNBH
3), place dark sections temperature to stir 15h, 300rpm, it is that the bag filter of 3500Da was dialysed vacuum 30Pa ,-45 ℃ 2 days in deionized water that solution is added the interception molecular weight, lyophilization 48h promptly gets 1: 1 chitosan of mol ratio-streptomycin covalent complex 121mg.
Measure through HPLC gel analysis system (production of Waters company), (25 ℃ of probe temperatures, mobile phase: the 3mmol sodium acetate, flow rate of mobile phase 0.5ml/min. standard substance adopt glucosan), the relative molecular weight of chitosan-streptomycin covalent complex is 7026Da, and qualification result as shown in Figure 1.
Embodiment 2
Get chitosan 150mg, streptomycin 2.714g is dissolved in respectively in the 40ml deionized water, and adds 1.16gNaCNBH
3, place dark sections temperature to stir 15h, 300rpm, it is that the bag filter of 3500Da was dialysed vacuum 30Pa ,-45 ℃ 2 days in deionized water that solution is added the interception molecular weight, lyophilization 48h promptly gets 1: 2 chitosan of mol ratio-streptomycin covalent complex 206mg.
The infrared instrument that gained chitosan-antibiotic covalent complex infrared spectrum adopts Bruker company to produce detects, and adopts pellet technique.(Dong Yanming; Wang Mian; Wu Yusong; Ruan Yonghong. cellulose science and technology 6,9 (2), 42-55 (2001)).Gained chitosan-streptomycin covalent complex infrared identification collection of illustrative plates as shown in Figure 2,2880～2900cm wherein
-1The absworption peak that occurs is an aldehyde radical remaining on the streptomycin, 1650cm
-1Be N-H, 1360～1250cm
-1Be C-N, 1153～1031cm
-1Be C-O.
Embodiment 3
Get chitosan 50mg, streptomycin 1.81g is dissolved in respectively in the 20ml deionized water, and adds 0.372g NaCNBH
3, place dark sections temperature to stir 15h, 300rpm, it is that the bag filter of 3500Da was dialysed vacuum 30Pa ,-45 ℃ 2 days in deionized water that solution is added the interception molecular weight, lyophilization 48h promptly gets 1: 4 chitosan of mol ratio-streptomycin covalent complex 71mg.
Gained chitosan-antibiotic covalent complex is dissolved in deuterium for water, and the nuclear magnetic resonance analyser that adopts Bruker company to produce detects.The nuclear magnetic resonance, NMR C of gained chitosan-streptomycin covalent complex composes as shown in Figure 3, and wherein 10ppm is the carbon atom on the methyl, and 30ppm is a methylene, is the sugar ring between 60～100ppm, and 160ppm is an aldehyde radical remaining on the streptomycin.
Embodiment 4
Get chitosan 50mg, streptomycin 2.72g is dissolved in respectively in the 20ml deionized water, and adds 0.372g NaCNBH
3, place dark sections temperature to stir 15h, 300rpm, it is that the bag filter of 3500Da was dialysed vacuum 30Pa ,-45 ℃ 2 days in deionized water that solution is added the interception molecular weight, lyophilization 48h promptly gets 1: 6 chitosan of mol ratio-streptomycin covalent complex 84mg.
Gained chitosan-antibiotic covalent complex is dissolved in deuterium for water, and the nuclear magnetic resonance analyser that adopts Bruker company to produce detects.The nuclear magnetic resonance, NMR H spectrum of described chitosan-streptomycin covalent complex as shown in Figure 4; wherein 1.0ppm is a methyl, and 2.0ppm is the carbonyl of deacetylation not on the chitosan, and 3.0～4.0ppm is the sugar ring; 4.5～5.0ppm is a water, 5.0～5.5ppm is the hydrogen on the carbon on the streptomycin.
Embodiment 5
Get chitosan 100mg, streptomycin 452.5mg is dissolved in respectively in the 30ml deionized water, and adds 0.744gNaCNBH
3Place dark sections temperature to stir 15h, 300rpm, interception molecular weight are that the bag filter of 3500Da was dialysed in deionized water 2 days, vacuum 30Pa, and-45 ℃, lyophilization 48 hours promptly gets 1: 0.5 chitosan of mol ratio-streptomycin chemical compound 100.3mg.
Embodiment 6
Get chitosan 50mg, streptomycin 4.525g is dissolved in respectively in the 30ml deionized water, and adds 0.372gNaCNBH
3Place dark sections temperature to stir 15h, 300rpm, interception molecular weight are that the bag filter of 3500Da was dialysed in deionized water 2 days, vacuum 30Pa, and-45 ℃, lyophilization 48 hours promptly gets 1: 10 chitosan of mol ratio-streptomycin chemical compound 60.3mg.
Embodiment 7
Get chitosan 50mg, acetylspiramycin 100mg is dissolved in the 40ml50% alcoholic solution, and adds 0.7gNaCNBH
3Place dark sections temperature to stir 15h, 300rpm, interception molecular weight are that the bag filter of 3500Da was dialysed in deionized water 2 days, vacuum 30Pa, and-45 ℃, lyophilization 48 hours promptly gets chitosan-acetylspiramycin chemical compound 46mg.
Embodiment 8
Get chitosan 50mg, clarithromycin 100mg is dissolved in the 60ml deionized water, and adds 0.672g NaCNBH
3Place dark sections temperature to stir 15h, 300rpm, interception molecular weight are that the bag filter of 3500Da was dialysed in deionized water 2 days, vacuum 30Pa, and-45 ℃, lyophilization 48 hours promptly gets chitosan-clarithromycin chemical compound 50.1mg.
Embodiment 9
Get chitosan 50mg, erythromycin 100mg is dissolved in the 40ml50% methanol, and adds 0.772g NaCNBH
3Place dark sections temperature to stir 15h, 300rpm, interception molecular weight are that the bag filter of 3500Da was dialysed in deionized water 2 days, vacuum 30Pa, and-45 ℃, lyophilization 48 hours promptly gets chitosan-erythromycin compounds 58.7mg.
Effect embodiment 1 chitosan-streptomycin covalent complex is to the inhibitory action of different tumor cells
Cell: gastric carcinoma cells SGC7901 (available from Shanghai medicine institute of the Chinese Academy of Sciences) and BGC823, human colon carcinoma HT29 (available from Shanghai medicine institute of the Chinese Academy of Sciences), human large intestine cancer Lovo cell, human liver cancer cell BEL-7402 (available from Shanghai medicine institute of the Chinese Academy of Sciences), SMMC-7721 and HepG2, human lung cancer cell A549 (available from Shanghai medicine institute of the Chinese Academy of Sciences), human esophagus cancer cell Eca-109, the Hela cell, application on human skin melanoma cell A375, human osteosarcoma cell MG63 (all available from the refined central laboratory in Central South University Hunan).
Medicine: chitosan-streptomycin covalent complex (chitosan-streptomycin mol ratio: 1: 4), medicine contrast chitosan (Qingdao cloud cosmos bio tech ltd), streptomycin (Aladdin).
Test method:
Tumor cell culture: RMPI1640 culture medium (available from Gibco company), 10% hyclone (available from Hyclone company), 100 μ g/ml streptomycins and and 100U/ml penicillin (all available from Gibco company), 5%CO
2, cultivate in 37 ℃ of CO2 gas incubator.
Collect the logarithmic (log) phase cell, adjust cell concentration to 10
5/ ml, every hole adds 100 μ l cell suspension.After placing incubator to cultivate 12h 96 orifice plates, every hole adds 90 μ l culture medium and 10 μ l different pharmaceuticals (drug level 1mg/ml), and every group to establish 6 holes parallel.5%CO
2, behind 37 ℃ of cultivation 24h, inhale and remove culture medium and medicine, add serum-free medium 100 μ l, every hole of while adds 10 μ l MTT solution (available from sigma company), and (5mg/ml is with 0.1mol/L PBS preparation, 0.22 μ m filters), continue to cultivate 4h, stop cultivating the centrifugal 5min of 3500rmp, the careful suction abandoned culture supernatant in the hole, every hole adds 100 μ l DMSO, and vibration 10min fully melts crystal.Under the 570nm wavelength, measure each hole absorbance value on microplate reader, the record result is an abscissa with time, and light absorption value is that vertical coordinate is drawn cell growth curve.The test triplicate.Three times the repeated trials result shows, chitosan-streptomycin covalent complex all has significant inhibition and toxic action to 12 kinds of human tumor cells, and its result is shown in Fig. 5 A-Fig. 5 L.
Effect embodiment 2 chitosans-streptomycin covalent complex different action times are to the inhibitory action of tumor cell
Cell: gastric carcinoma cells SGC7901
Medicine: chitosan-streptomycin covalent complex (chitosan-streptomycin mol ratio: 1: 4), medicine contrast chitosan (Qingdao cloud cosmos bio tech ltd), streptomycin (Aladdin).
Test method:
Stomach cancer cell is cultivated: RMPI1640 culture medium, 10% hyclone, 100 μ g/ml streptomycins and 100U/ml penicillin, 5%CO
2, cultivate in 37 ℃ of CO2 gas incubator.
Collect the logarithmic (log) phase cell, adjust cell concentration to 10
5/ ml, every hole adds 100 μ l cell suspension.After placing incubator to cultivate 12h 96 orifice plates, every hole adds 90 μ l culture medium and 10 μ l different pharmaceuticals (drug level 1mg/ml), and every group to establish 6 holes parallel.5%CO
2, cultivate 6h, 12h respectively for 37 ℃, behind the 24h, inhale and remove culture medium and medicine, add serum-free medium 100 μ l, every hole adds 10 μ l MTT solution (5mg/ml, with 0.1mol/L PBS preparation, 0.22 μ m filters) simultaneously, continue to cultivate 4h, stop cultivating the centrifugal 5min of 3500rmp, the careful suction abandoned culture supernatant in the hole, every hole adds 100 μ l DMSO, and vibration 10min fully melts crystal.Under the 570nm wavelength, measure each hole absorbance value on microplate reader, the record result is an abscissa with time, and light absorption value is that vertical coordinate is drawn cell growth curve.The experiment triplicate, experimental result is shown in Fig. 6 A-Fig. 6 C.
Above-mentioned experimental result shows, the drug effect different time (6h, 12h, 24h) after, chitosan-streptomycin covalent complex (1: 4) has significant inhibition and toxic action to tumor cell (gastric carcinoma cells SGC7901).
Effect embodiment 3 different proportions chitosan-streptomycin covalent complex is to the inhibitory action of tumor cell
Cell: human lung cancer cell A549
Medicine: chitosan-streptomycin covalent complex (chitosan-streptomycin mol ratio: 1: 1,1: 2,1: 4,1: 6), medicine contrast chitosan (Qingdao cloud cosmos bio tech ltd), streptomycin (Aladdin).
Test method:
Lung carcinoma cell is cultivated: RMPI1640 culture medium, 10% hyclone, 100 μ g/ml streptomycins and 100U/ml penicillin, 5%CO
2, cultivate in 37 ℃ of CO2 gas incubator.
Collect the logarithmic (log) phase cell, adjust cell concentration to 10
5/ ml, every hole adds 100 μ l cell suspension.After placing incubator to cultivate 12h 96 orifice plates, every hole adds 90 μ l culture medium and 10 μ l different pharmaceuticals (drug level 1mg/ml), and every group to establish 6 holes parallel.5%CO
2, behind 37 ℃ of cultivation 24h, inhale and remove culture medium and medicine, add serum-free medium 100 μ l, every hole of while adds 10 μ l MTT solution, and (5mg/ml is with 0.1mol/L PBS preparation, 0.22 μ m filters), continue to cultivate 4h, stop cultivating the centrifugal 5min of 3500rmp, the careful suction abandoned culture supernatant in the hole, every hole adds 100 μ l DMSO, and vibration 10min fully melts crystal.Under the 570nm wavelength, measure each hole absorbance value on microplate reader, the record result is an abscissa with time, and light absorption value is that vertical coordinate is drawn cell growth curve.The experiment triplicate, experimental result is shown in Fig. 7 A-Fig. 7 D.Above-mentioned experimental result shows, and the chitosan of different proportion-streptomycin covalent complex (1: 1,1: 2,1: 4,1: 6) tumor cell (human lung cancer cell A549) there are obvious suppression and toxic action, action effect increases along with the increase of streptomycin ratio.
Wherein chitosan and streptomycin mol ratio are that chitosan-streptomycin covalent complex tumor suppression of 1: 4 is effective, and the use amount of streptomycin is lower.
Effect embodiment 4 different chitosans-antibiotic covalent complexes are to the inhibitory action of tumor cell
Cell: human lung cancer cell A549
Medicine: chitosan-acetylspiramycin covalent complex (chitosan-streptomycin mol ratio: 1: 1), chitosan-gentamycin covalent complex, chitosan-kanamycin covalent complex, medicine contrast chitosan (Qingdao cloud cosmos bio tech ltd), acetylspiramycin (Aladdin), clarithromycin, erythromycin.
Test method:
Lung carcinoma cell is cultivated: RMPI1640 culture medium, 10% hyclone, 100 μ g/ml streptomycins and 100U/ml penicillin, 5%CO
2, cultivate in 37 ℃ of CO2 gas incubator.
Collect the logarithmic (log) phase cell, adjust cell concentration to 10
5/ ml, every hole adds 100 μ l cell suspension.After placing incubator to cultivate 12h 96 orifice plates, every hole adds 90 μ l culture medium and 10 μ l different pharmaceuticals (drug level 1mg/ml), and every group to establish 6 holes parallel.5%CO
2, behind 37 ℃ of cultivation 24h, inhale and remove culture medium and medicine, add serum-free medium 100 μ l, every hole of while adds 10 μ l MTT solution, and (5mg/ml prepares with 0.1mol/LPBS, 0.22 μ m filters), continue to cultivate 4h, stop cultivating the centrifugal 5min of 3500rmp, the careful suction abandoned culture supernatant in the hole, every hole adds 100 μ l DMSO, and vibration 10min fully melts crystal.Under the 570nm wavelength, measure each hole absorbance value on microplate reader, the record result is an abscissa with time, and light absorption value is that vertical coordinate is drawn cell growth curve.The experiment triplicate, experimental result as shown in Figure 8.Above-mentioned experimental result shows that different chitosans-antibiotic covalent complex has obvious suppression and toxic action to tumor cell (human lung cancer cell A549).
Should be understood that, for those of ordinary skills, can be improved according to the above description or conversion, and all these improvement and conversion all should belong to the protection domain of claims of the present invention.
Claims (8)
1. the preparation method of antibiotic-chitosan covalent complex is characterized in that, may further comprise the steps:
(1) preparation chitosan and antibiotic aqueous solution are mixing in 1: 0.5～1: 10 with chitosan and antibiotic according to molar ratio, add NaCNBH
3, lucifuge stirs, and 15 ℃～30 ℃ were reacted 15～72 hours;
(2) aqueous solution of step (1) gained being moved into molecular cut off is in 1400～10000 the bag filter, to dialyse 1～5 day, obtains dialysis solution;
(3) with the dialysis solution drying of step (2) gained, promptly.
2. preparation method as claimed in claim 1 is characterized in that, the described antibiotic of step (1) is gentamycin or streptomycin or kanamycin.
3. preparation method as claimed in claim 1 is characterized in that, described chitosan of step (1) and antibiotic molar ratio are 1: 1～1: 6.
4. preparation method as claimed in claim 1 is characterized in that, the described NaCNBH of step (1)
3Addition be 30%～50% of described antibiotic quality.
5. preparation method as claimed in claim 1 is characterized in that, the described bag filter molecular cut off of step (2) is 3000～5000, and the time of described dialysis is 2～5 days.
6. preparation method as claimed in claim 1 is characterized in that, the described exsiccant mode of step (3) is vacuum lyophilization, vacuum drying, spray drying, oven dry or infrared drying.
One kind as claim 1～6 as described in each preparation method prepare the chitosan-antibiotic covalent complex of gained.
8. the application of chitosan as claimed in claim 7-antibiotic covalent complex in the preparation antitumor drug.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105497048B (en) * | 2015-12-17 | 2018-05-04 | 西北农林科技大学 | A kind of preparation and its application of proteoglycans antibiotic complex |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1376735A (en) * | 2001-03-22 | 2002-10-30 | 贺利氏古萨两合有限公司 | Antibiotics-polymer composition |
-
2013
- 2013-04-19 CN CN2013101514421A patent/CN103212087A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1376735A (en) * | 2001-03-22 | 2002-10-30 | 贺利氏古萨两合有限公司 | Antibiotics-polymer composition |
Non-Patent Citations (2)
| Title |
|---|
| V.K.MOURYA,ET AL: "Chitosan-modifications and applications:Opportunities galore", 《REACTIVE & FUNCTIONAL POLYMERS》 * |
| YALPANI,ET AL: "Some Chemical and Analytical Aspects of Polysaccharide Modifications.3.Formation of Branched-Chain,Soluble Chitosan Derivatives", 《MACROMOLECULES》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105497048B (en) * | 2015-12-17 | 2018-05-04 | 西北农林科技大学 | A kind of preparation and its application of proteoglycans antibiotic complex |
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Application publication date: 20130724 |