CN103784968A - Chitosan-antibiotic covalent compound and preparation method and application thereof - Google Patents
Chitosan-antibiotic covalent compound and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a chitosan-antibiotic covalent compound and a preparation method and application thereof. The preparation method comprises the following steps: (1) preparing chitosan and an antibiotic in the molar ratio of 1:0.1-1:10, dissolving into water, adding NaCNBH3, stirring away from light, and reacting at 0-90 DEG C for 0.5-72 hours; (2) dialyzing an obtained mixed solution for 1-5 days; (3) drying the obtained dialysate to obtain the chitosan-antibiotic covalent compound. Raw materials for preparing the chitosan-antibiotic covalent compound are readily available, and a synthetic method is simple and convenient. The covalent compound has a strong destructive effect on formed bacterial biofilms, is wide in sterilizing range and definite in effective components, and has a good inhibiting effect on the formation of biofilms.
Description
Technical field
The invention belongs to pharmaceutical preparations technology field, be specifically related to a kind of chitosan-antibiotic covalent complex and its preparation method and application.
Background technology
Biomembrane (biofilms, BF) is the special group of one that the extracellular macromole polymer of microbial cell (as antibacterial, fungus, protozoon) and generation thereof is formed, and is highly organized multi-cellular structure.Bacterial biof iotalm is antibacterial is adsorbed in life or the formation of non-life body surface in growth course a kind of growth pattern for adapting to living environment, is made up of the extracellular matrix of antibacterial and self secretion.Research worker can form the biomembranous antibacterial of this class by these and be called biomembrane antibacterial (BF antibacterial).BF can see the impact of bacterial biof iotalm in nature, some industrial environment (as fields such as water-carriage system, food processing, shippings), humans and animals body.In addition, the relation of BF and multiple human body related infection disease is also very close.In the time that antibacterial exists with BF form, drug resistance obviously strengthens (10～1000 times), and antibiotic is become to insensitive, and inducible resistance produces; Can also resist host's defense system, the surface that grows in tooth, gingiva, skin, lung, urethra and other organs at premorbid reaches even several years several months, causes refractory infection, brings and has a strong impact on to clinical treatment.According to estimates, 65% human infection is relevant with biomembrane.The resistance mechanism of BF is different from the bacterium that swims, and the antibacterials of valid density can be killed rapidly the swim antibacterial of growth and the antibacterial on BF surface, but the antibacterial of BF depths is difficult to effectively kill.Because traditional antibiotic is as not good enough in the infection curative effect that streptomycin, gentamycin, penicillin cause antibacterial BF, therefore, be badly in need of clinically finding the medicine that can kill BF depths antibacterial.
Chitosan (chitosan) is by the nature complex that extensively chitin of existence obtains through deacetylation, extensive use in medicine, food, chemical industry, cosmetics, water treatment, METAL EXTRACTION and recovery, biochemistry and biomedical engineering.Streptomycin (streptomycin) be after penicillin second produce and for clinical a kind of glucosamine type antibiotic, streptomycin application is clinically very extensive.
Summary of the invention
Therefore, technical problem to be solved by this invention is for lacking clinically the medicine that can effectively kill biomembrane depths antibacterial, the infection curative effect that tradition antibiotic causes bacterial biof iotalm is not good enough, and can not stop the film formed problem of bacterium living beings, a kind of chitosan-antibiotic covalent complex and its preparation method and application is provided.
For solving the problems of the technologies described above, one of technical scheme that the present invention takes is: a kind of preparation method of chitosan-antibiotic covalent complex, and described preparation method comprises the following steps:
(1) preparing chitosan and antibiotic aqueous solution, is that 1:0.1～1:10 mixes by chitosan and antibiotic according to molar ratio, adds NaCNBH
3, lucifuge stirs, and 0 ℃～90 ℃ are reacted 0.5～72 hour;
(2) aqueous solution of step (1) gained is moved in the bag filter that molecular cut off is 1000～14000, dialyse 1～5 day, obtain dialysis solution;
(3) dialysis solution of step (2) gained is dry, to obtain final product.
Wherein the described chitosan of step (1) is the conventional chitosan in this area.Described chitosan (chitosan) is that the chitin (chitin) extensively being existed by nature obtains through deacetylation, and chemical name is Chitosan (1-4)-2-amino-B-D glucose.Wherein said antibiotic is the antibiotic of this area routine.Described antibiotic is preferably aminoglycoside antibiotics.Described aminoglycoside antibiotics is the glucosides class antibiotic being formed by connecting by oxo bridge by amino sugar and aminocyclitol.Described antibiotic is preferably streptomycin, kanamycin or gentamycin.Antibiotic of the present invention is preferably streptomycin.
Wherein the described chitosan of step (1) and antibiotic aqueous solution form mixed liquor, and described chitosan and antibiotic mol ratio are preferably 1:0.1～1:10, are more preferably 1:0.5～1:6, are 1:0.5 best.The time of described reaction is preferably 0.5～72 hour, is more preferably 10～48 hours, is 24 hours best.The temperature of described reaction is preferably 0 ℃～90 ℃, is more preferably 15 ℃～30 ℃, is best 20 ℃.
The wherein described NaCNBH of step (1)
3addition be preferably 30%～50% of described antibiotic content, addition is 30% best, 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 through film, can be used for the solute that isolated molecule amount varies in size, the material of holding back threshold value molecular weight lower than film can diffuse through film, and the material of holding back threshold value molecular weight higher than film is retained in the opposite side of semipermeable membrane.Wherein said bag filter is this area conventional dialysis bag.The molecular cut off of described bag filter is preferably 1000～14000, is more preferably 3000～5000, and molecular cut off is 3500 best.The time of wherein said dialysis is preferably 2～5 days, is more preferably 2～3 days, and dialysis time is 2 days best.
What wherein step (3) was described is dried as this area conventional drying mode.Described dry preferably for vacuum lyophilization, vacuum drying, spraying are dry, oven dry or infrared drying, be vacuum lyophilization best.The parameter of described vacuum lyophilization is preferably: 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: preparation method as above is prepared chitosan-antibiotic covalent complex of gained.
In described chitosan-antibiotic covalent complex, chitosan and antibiotic mol ratio are preferably 1:0.1～1:10, are more preferably 1:0.5～1:6, are 1:0.5 best.
For solving the problems of the technologies described above, three of the technical scheme that the present invention takes is: chitosan-antibiotic covalent complex as above is in the purposes of preparing in anti-infectives.
Purposes of the present invention preferably refers to using chitosan-antibiotic covalent complex as active component, makes the anti-infectives of various dosage forms with pharmaceutically acceptable adjuvant.Wherein said pharmaceutically acceptable adjuvant is the conventional adjuvant using in this area.Described excipient substance refers to excipient and additives used while producing medicine and prescription being dispensed.
Wherein said anti-infectives is more preferably bacterial-infection resisting class medicine.Described antibacterial is the conventional pathogenic bacterium in this area.Described antibacterial is preferably gram positive bacteria and gram negative bacteria.Wherein gram positive bacteria is preferably Listeria monocytogenes, and wherein gram negative bacteria is preferably staphylococcus aureus or Pseudomonas aeruginosa.
Meeting on the basis of this area general knowledge, above-mentioned each optimum condition, can combination in any, obtains the preferred embodiments of the invention.
Agents useful for same of the present invention and raw material be commercially available obtaining all.
Positive progressive effect of the present invention is: the raw material of preparing chitosan-antibiotic covalent complex of the present invention is easy to get, and simple synthetic method is easily gone; Prove through microbiological test, this chitosan-antibiotic covalent complex has stronger destruction to the bacterial biof iotalm having formed, and bactericidal range is wide, and effective ingredient is clear and definite; This complex has good inhibitory action to biomembranous formation.Meanwhile, described chitosan-antibiotic covalent complex can effectively reduce the drug resistance producing when antibacterial exists with biomembrane form, improves antibacterial to the antibiotic sensitivity of tradition, reduces traditional antibiotic use amount.
Accompanying drawing explanation
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 destruction design sketch of chitosan-streptomycin covalent complex to bacterial biof iotalm; Note: contrast the blank for not adding any medicine, L is streptomycin, and CS is chitosan, and L+CS is the mixture of streptomycin and chitosan.A represents the single Liszt of increasing antibacterial; B represents staphylococcus aureus; C represents Aerugo bacillus.
Fig. 6 is the inhibitory action design sketch of chitosan-streptomycin covalent complex to antibacterial in biomembrane; Note: contrast the blank for not adding any medicine, L is streptomycin, and CS is chitosan, and L+CS is the mixture of streptomycin and chitosan, A represents the single Liszt of increasing antibacterial; B represents staphylococcus aureus; C represents Aerugo bacillus.
Fig. 7 is the inhibitory action design sketch of chitosan-streptomycin covalent complex to antibacterial in biomembrane; Note: contrast the blank for not adding any medicine, L is streptomycin, and CS is chitosan, and L+CS is the mixture of streptomycin and chitosan.A represents the single Liszt of increasing antibacterial; B represents staphylococcus aureus; C represents Aerugo bacillus.
Fig. 8 is the fungistatic effect figure of chitosan-streptomycin covalent complex in biofilm formation process; Note: contrast the blank for not adding any medicine, L is streptomycin, and CS is chitosan, and L+CS is the mixture of streptomycin and chitosan.A represents the single Liszt of increasing antibacterial; B represents staphylococcus aureus; C represents Aerugo bacillus.
Fig. 9 is the fungistatic effect figure of chitosan-streptomycin covalent complex in biofilm formation process; Note: contrast the blank for not adding any medicine, L is streptomycin, and CS is chitosan, and L+CS is the mixture of streptomycin and chitosan.A represents the single Liszt of increasing antibacterial; B represents staphylococcus aureus; C represents Aerugo bacillus.
The specific embodiment
Mode below by embodiment further illustrates the present invention, but does not therefore limit the present invention among described scope of embodiments.Agents useful for same of the present invention and raw material be commercially available obtaining all.The experimental technique of unreceipted actual conditions in the following example, according to conventional method and condition, or selects according to catalogue.
Get chitosan 50mg, streptomycin 45.3mg is dissolved in respectively in 20ml deionized water, and add 0.372g sodium cyanoborohydride (NaCNBH3), be placed in 300rpm under the 15 ℃ of conditions in dark place, stir 10h, solution is added to the bag filter that interception molecular weight is 3000, in deionized water, dialyse 2 days, vacuum 20pa ,-35 ℃ of lyophilizations 48 hours, obtain chitosan-streptomycin complex 85.8mg of mol ratio 1:0.1.Measure through HPLC gel analysis system (production of Waters company), (25 ℃ of probe temperatures, mobile phase: 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 100mg, streptomycin 452.5mg is dissolved in respectively in 30ml deionized water, and add 0.744g NaCNBH3 to be placed in 20 ℃, dark place, 300rpm, stirs 24h, and solution is added to the bag filter that interception molecular weight is 3500, in deionized water, dialyse 2 days, vacuum 30Pa ,-45 ℃ of lyophilizations 48 hours, obtain chitosan-streptomycin covalent complex 497.3mg of mol ratio 1:0.5.Gained chitosan-antibiotic covalent complex infrared spectrum adopts the infrared instrument that Bruker company produces to detect, and adopts pellet technique.(Dong Yanming; Wang Mian; Wu Yusong; Ruan Yonghong. Cellulose Science technology 6,9 (2), 42-55 (2001)).Gained chitosan-streptomycin covalent complex infrared identification collection of illustrative plates as shown in Figure 2, wherein 2880～2900cm
-1the absworption peak occurring is aldehyde radical remaining on streptomycin, 1650cm
-1n-H, 1360 ~ 1250cm
-1c-N, 1153 ~ 1031cm
-1for C-O.
Embodiment 3
Get chitosan 150mg, streptomycin 2.714g is dissolved in respectively in 40ml deionized water, and adds 1.16g NaCNBH3, is placed in 25 ℃, dark place, and 300rpm, stirs 36h, and solution is added to the bag filter that interception molecular weight is 4000, dialyses 3 days in deionized water.At vacuum 20Pa ,-45 ℃ of lyophilizations 48 hours, obtain chitosan-streptomycin covalent complex 2.55g of mol ratio 1:2.Gained chitosan-antibiotic covalent complex is dissolved in to deuterated water, adopts the nuclear magnetic resonance analyser that Bruker company produces to detect.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 methyl, and 30ppm is methylene, is sugar ring between 60 ~ 100ppm, and 160ppm is aldehyde radical remaining on streptomycin.
Embodiment 4
Get chitosan 50mg, streptomycin 1.81g is dissolved in respectively in 20ml deionized water, and add 0.372g NaCNBH3, be placed in 300rpm under the 30 ℃ of conditions in dark place, stir 48h, solution is added to the bag filter that interception molecular weight is 4500, in deionized water, dialyse 4 days, vacuum 30Pa ,-45 ℃ of lyophilizations 48 hours, obtain chitosan-streptomycin covalent complex 1.66g of mol ratio 1:4.Gained chitosan-antibiotic covalent complex is dissolved in to deuterated water, adopts the nuclear magnetic resonance analyser that Bruker company produces to detect.The nuclear magnetic resonance, NMR H of described chitosan-streptomycin covalent complex composes as shown in Figure 4; wherein 1.0ppm is methyl, and 2.0ppm is the carbonyl of deacetylation not on chitosan, and 3.0 ~ 4.0ppm is sugar ring; 4.5 ~ 5.0ppm is water, and 5.0 ~ 5.5ppm is the hydrogen on the carbon on streptomycin.
Embodiment 5
Get chitosan 50mg, streptomycin 2.72g is dissolved in respectively in 20ml deionized water, and add 0.372g NaCNBH3, be placed in 25 ℃, dark place, 300rpm, stir 48h, solution is added to the bag filter that interception molecular weight is 5000, in deionized water, dialyse 5 days, vacuum 30Pa,-40 ℃ of lyophilizations 36 hours, obtain chitosan-streptomycin covalent complex 2.51g of mol ratio 1:6.
Embodiment 6
Get chitosan 150mg, streptomycin 13.75g is dissolved in respectively in 20ml deionized water, and add 6.785g NaCNBH3, be placed in 0 ℃, dark place, 300rpm, stirs 72h, solution is added to the bag filter that interception molecular weight is 14000, in deionized water, dialyse 5 days, dry 36 hours, obtain chitosan-streptomycin covalent complex 12.28g of mol ratio 1:10.
Embodiment 7
Get chitosan 50mg, kanamycin 58.3mg is dissolved in respectively in 20ml deionized water, and add 17.49mg sodium cyanoborohydride (NaCNBH3), be placed in 300rpm under the 90 ℃ of conditions in dark place, stir 72h, solution is added to the bag filter that interception molecular weight is 14000, in deionized water, dialyse 1 day, infrared drying 48 hours, obtains chitosan-kanamycin complex 85.8mg of mol ratio 1:0.1.
Get chitosan 50mg, streptomycin 2.72g is dissolved in respectively in 20ml deionized water, and add 0.816g NaCNBH3, be placed in 25 ℃, dark place, 300rpm, stirs 48h, solution is added to the bag filter that interception molecular weight is 5000, in deionized water, dialyse 5 days, spraying is dry, obtains chitosan-streptomycin covalent complex 2.51g of mol ratio 1:6.
Embodiment 9
Get chitosan 150mg, streptomycin 2.714g is dissolved in respectively in 40ml deionized water, and adds 0.81g NaCNBH3, is placed in 60 ℃, dark place, and 300rpm, stirs 0.5h, and solution is added to the bag filter that interception molecular weight is 1000, dialyses 1 day in deionized water.Gained dialysis solution, at vacuum 20Pa, is dried 48 hours, obtains chitosan-streptomycin covalent complex 2.55g of mol ratio 1:2.
The destruction of effect embodiment 1 chitosan-streptomycin covalent complex to bacterial biof iotalm
Microorganism: Listeria monocytogenes CMCC 54004, staphylococcus aureus ATCC 29213, Pseudomonas aeruginosa ATCC9027.
Medicine: embodiment 1-9 prepares gained chitosan-streptomycin covalent complex, medicine contrast chitosan (Yun Zhou bio tech ltd, Qingdao), streptomycin (Aladdin).
Test method: add 0.1ml containing 5 × 10 in the 96 each holes of well culture plate
8～10
9antibacterial, put into 37 ℃ of incubators and cultivate 24 hours, allow antibacterial note wall form biomembrane.Suck culture fluid, with PBS(0.1mol/L) to wash three times, every hole adds 90 μ l LB culture medium and 10 μ l medicines.Medicine is that (wherein the mol ratio of chitosan and streptomycin is respectively 1:0.1 to chitosan-streptomycin covalent complex, 1:0.5,1:2,1:4,1:6,1:10), chitosan (CS), streptomycin (L), and the mixture of chitosan and streptomycin (CS+L) (wherein chitosan and streptomycin mass ratio are 1:1), described drug level is 0.25mg/ml.Every group of parallel 6 holes.Be placed in 37 ℃ of incubators and cultivate 24h.Suck medicine, with PBS(0.1mol/L) carefully to wash three times, every hole adds 0.1ml 99% methanol solution fixation of bacteria 15 minutes.Suck methanol solution, every hole adds 0.1ml 0.1% crystal violet dye liquor, places 20 minutes in room temperature.Get rid of gently dyeing liquor, with the each hole of distilled water wash, culture plate is inverted in to suck dry moisture in absorbent paper.Natural drying or 37 ℃ of oven dry.Before mensuration, every hole adds 0.1ml 33% acetic acid decolouring, fully after vibration, measures absorbance at 595nm place.Test in triplicate.
Three times repeated trials result shows, the biomembrane that chitosan-streptomycin covalent complex forms above-mentioned three kinds of antibacterials has significant inhibition, destruction, and all has significant effect in 24h, and its result is as shown in Fig. 5 (A, B, C).
The inhibitory action of effect embodiment 2 chitosans-streptomycin covalent complex to biomembrane depths antibacterial
Bacterial strain: with effect embodiment 1.
Medicine: with effect embodiment 1.
Test method: add 0.1ml containing 5 × 10 in the 96 each holes of well culture plate
8～10
9antibacterial, put into 37 ℃ of incubators and cultivate 24 hours, allow antibacterial note wall form biomembrane.Suck culture fluid, with PBS(0.1mol/L) carefully to wash three times, every hole adds the medicine (with effect embodiment 1) that 90 μ l culture medium are different with 10 μ l, every group of parallel 6 holes.Be placed in 37 ℃ of incubators and cultivate 24h.Suck medicine, with PBS(0.1mol/L) carefully wash three times, every hole adds 90 μ l culture medium and 10 μ l MTT solution (5mg/ml PBS prepares, and 0.22 μ m filters), continues to hatch 3h, stop cultivating, the centrifugal 5min of 3500rmp, careful suction abandoned culture supernatant in hole, and every hole adds 100 μ l DMSO, vibration 10min, fully melts crystal.Under 490nm wavelength, measure each hole absorbance value on enzyme linked immunological monitor, record result, take the time as abscissa, light absorption value is that vertical coordinate is drawn cell growth curve.Test in triplicate.
Three times repeated trials result shows, in 24h, chitosan-streptomycin covalent complex has significant inhibition, killing effect to above-mentioned three kinds of antibacterials of biomembrane depths, and its result is as shown in Fig. 6 (A, B, C).
The inhibitory action of effect embodiment 3 chitosans-streptomycin covalent complex to biomembrane depths antibacterial
Bacterial strain: with effect embodiment 1.
Medicine: with effect embodiment 1.
In the 96 each holes of well culture plate, add 0.1ml containing 5 × 10
8～10
9antibacterial, put into 37 ℃ of incubators and cultivate 24 hours, allow antibacterial note wall form biomembrane.Suck culture fluid, with the careful washing of PBS three times, every hole adds 90 μ l culture medium and 10 μ l medicines (with effect embodiment 1), every group of parallel 6 holes.Be placed in 37 ℃ of incubators and cultivate 24h.Suck medicine, with the careful washing of PBS three times, add after 0.1ml0.2%Triton X-100 cracking 1h dilution point plate, counting.
Result of the test shows, chitosan-streptomycin covalent complex of different mol ratio example above-mentioned three kinds of antibacterials to biomembrane depths in 24h have antibacterial, bactericidal effect significantly, and its result is as shown in Fig. 7 (A, B, C).
The bactericidal action of effect embodiment 4 chitosans-streptomycin covalent complex in biofilm formation process
Bacterial strain: with effect embodiment 1.
Medicine: with effect embodiment 1.
Test method: add 90 μ l containing 5 × 10 in the 96 each holes of well culture plate
8～10
9the solution of antibacterial, add the medicine that 10 μ l are different (with effect embodiment 1) simultaneously, every group of parallel 6 holes, put into 37 ℃ of incubators and cultivate 24 hours.Suck medicine, with PBS(0.1mol/L) carefully to wash three times, every hole adds 90 μ l culture medium and 10 μ l MTT solution, and (concentration is 5mg/ml, prepares with PBS, 0.22 μ m filters), continue to hatch 3h, stop cultivating the centrifugal 5min of 3500rmp, careful suction abandoned culture supernatant in hole, every hole adds 100 μ l DMSO, and vibration 10min, fully melts crystal.Under 490nm wavelength, measure each hole absorbance value on enzyme linked immunological monitor, record result, take the time as abscissa, light absorption value is that vertical coordinate is drawn cell growth curve.Test in triplicate.
Result of the test shows, chitosan-streptomycin covalent complex, in the 24h of described three kinds of bacterial biof iotalm forming processes, has significant antibacterial and bactericidal action, and its result is as shown in Fig. 8 (A, B, C).Chitosan-streptomycin covalent complex bactericidal effect that wherein chitosan and streptomycin mol ratio are 1: 0.5 is better, and the use amount of streptomycin is lower.
The bactericidal action of effect embodiment 5 chitosans-streptomycin covalent complex in biofilm formation process
Bacterial strain: with effect embodiment 1.
Medicine: with effect embodiment 1.
Test method: add 90 μ l containing 5 × 10 in the 96 each holes of well culture plate
8～10
9antibacterial, add the medicine that 10 μ l are different (with effect embodiment 1) simultaneously, every group of parallel 6 holes, put into 37 ℃ of incubators and cultivate 24 hours.Suck liquid in 96 orifice plates, with PBS(0.1mol/L) carefully wash three times, add after 0.1ml 0.2%Triton X-100 cracking 1h dilution point plate, counting.
Result of the test shows, chitosan-streptomycin covalent complex, in the 24h of above-mentioned three kinds of bacterial biof iotalm forming processes, has significant antibacterial and bactericidal action, and its result is as shown in Fig. 9 (A, B, C).Chitosan-streptomycin covalent complex bactericidal effect that wherein chitosan and streptomycin mol ratio are 1: 0.5 is good, and the use amount of streptomycin is lower.
Should be understood that, after having read foregoing of the present invention, those skilled in the art can make various changes or modifications the present invention, these equivalent form of values fall within the application's appended claims limited range equally.
Claims (10)
1. a preparation method for chitosan-antibiotic covalent complex, is characterized in that, this preparation method comprises the following steps:
(1) preparing chitosan and antibiotic aqueous solution, is that 1:0.1～1:10 mixes by chitosan and antibiotic according to molar ratio, adds NaCNBH
3, lucifuge stirs, and 0 ℃～90 ℃ are reacted 0.5～72 hour;
(2) aqueous solution of step (1) gained is moved in the bag filter that molecular cut off is 1000～14000, dialyse 1～5 day, obtain dialysis solution;
(3) dialysis solution of step (2) gained is dry, to obtain final product.
2. preparation method as claimed in claim 1, is characterized in that, the described antibiotic of step (1) is gentamycin, streptomycin or kanamycin.
3. preparation method as claimed in claim 1, is characterized in that, the described antibiotic of step (1) is streptomycin.
4. preparation method as claimed in claim 1, is characterized in that, the described chitosan of step (1) and antibiotic molar ratio are 1:0.5～1:6.
5. preparation method as claimed in claim 1, is characterized in that, the described NaCNBH of step (1)
3addition be 30%～50% of described antibiotic content, described percentage ratio is mass percent.
6. preparation method as claimed in claim 1, is characterized in that, the temperature of the described reaction of step (1) is 15 ℃～30 ℃, and the time of described reaction is 10～48 hours.
7. 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.
8. preparation method as claimed in claim 1, is characterized in that, the described dry mode of step (3) is that vacuum lyophilization, vacuum drying, spraying are dry, oven dry or infrared drying.
9. the preparation method as described in claim 1～8 any one is prepared chitosan-antibiotic covalent complex of gained.
10. chitosan-antibiotic covalent complex as claimed in claim 9 is in the purposes of preparing in anti-infectives.
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| CN106596922A (en) * | 2016-12-06 | 2017-04-26 | 湖南大学 | Detection agent used for detecting kanamycin and preparation method and application thereof |
| 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 |
| CN108744014A (en) * | 2018-06-14 | 2018-11-06 | 福州大学 | A kind of preparation method and products thereof with slow releasing function antiseptic dressing |
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| CN101700243A (en) * | 2009-11-20 | 2010-05-05 | 佘振定 | Novel medicine carrying woundplast with treating function |
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| CN101700243A (en) * | 2009-11-20 | 2010-05-05 | 佘振定 | Novel medicine carrying woundplast with treating function |
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| MANSUR YALPANI ET AL.: "Some Chemical and Analytical Aspects of Polysaccharide Modifications.3.Formation of Branched-Chain, Soluble Chitosan Derivatives", 《MACROMOLECULES》, vol. 17, 31 December 1984 (1984-12-31), pages 280 * |
Cited By (5)
| 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 |
| CN106596922A (en) * | 2016-12-06 | 2017-04-26 | 湖南大学 | Detection agent used for detecting kanamycin and preparation method and application thereof |
| CN106596922B (en) * | 2016-12-06 | 2018-06-15 | 湖南大学 | Detection reagent for detecting kanamycins and its preparation method and application |
| CN108744014A (en) * | 2018-06-14 | 2018-11-06 | 福州大学 | A kind of preparation method and products thereof with slow releasing function antiseptic dressing |
| CN108744014B (en) * | 2018-06-14 | 2021-04-27 | 福州大学 | Preparation method of antibacterial dressing with slow release effect and product thereof |
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