CN108329467B - Preparation method of hyperbranched antibacterial peptide polymer - Google Patents
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Abstract
The invention discloses a preparation method of a hyperbranched antibacterial peptide polymer, which comprises the following steps: 1) and synthesis of amino-terminated hyperbranched polyamidoamine: preparing amino-terminated low molecular weight hyperbranched polyamidoamine by Michael addition reaction of methyl acrylate and diethylenetriamine; 2) lysine-N-carboxylic anhydride and valine-N-carboxylic anhydride synthesis: using hyperbranched polyamidoamine as a core, and respectively reacting benzyloxycarbonyl lysine and valine with triphosgene to obtain lysine-N-carboxylic anhydride and valine-N-carboxylic anhydride; 3) and (3) synthesizing a hyperbranched antibacterial peptide polymer: the lysine-N-carboxylic anhydride and the valine-N-carboxylic anhydride are grafted to the terminal amino group of the hyperbranched polyamidoamine through a polypeptide chain formed by ring-opening polymerization, and the antibacterial peptide polymer with the hyperbranched structure is prepared after amino deprotection and dialysis. The invention has the advantages of simple preparation method, easily obtained raw materials, lower synthesis cost and convenient large-scale production.
Description
Technical Field
The invention belongs to the technical field of preparation of antibacterial biomaterials, and particularly relates to a preparation method of a hyperbranched antibacterial peptide polymer.
Background
The generation of drug-resistant superbacteria due to the abuse of antibiotics in recent years has seriously threatened the life safety of human beings. The antibacterial peptide is a kind of antimicrobial and some malignant cell short peptide produced by organism in the defense reaction against pathogenic microorganism. Because the natural biological antibacterial peptide is a part of an immune system of eukaryote, the natural biological antibacterial peptide has broad-spectrum antibacterial property, and especially has a killing effect on multiple drug-resistant bacteria, and the antibacterial activity has the characteristics of high efficiency, stability, rapidness, difficult generation of resistance and the like, the development and utilization of the natural biological antibacterial peptide is expected to become a new way for people to get rid of the drug-resistant bacteria crisis and the anti-tumor treatment, and therefore, the potential application value of the antibacterial peptide is widely concerned by scholars. Although the biological antibacterial peptide has a wide prospect in controlling infection, the practical clinical application is greatly limited. The production cost of bio-antimicrobial peptides is extremely high compared to traditional antibiotics, which is a major obstacle limiting their widespread use. In addition, larger concentrations of the biological antimicrobial peptides induce toxicity in mammalian cells.
The cationic polymer can show higher antibacterial activity through selective design, is relatively cheap to synthesize, and can realize large-scale production. These antimicrobial polymers are typically designed with cationic and hydrophobic groups dispersed in the polymer backbone or with alkylated quaternary ammonium groups on long hydrophobic chains. It has been found that a polymerized peptide having N-carboxyanhydride cation (lysine) and hydrophobic (alanine, phenylalanine, leucine) amino acid residues, which has biodegradability and antibacterial activity similar to those of a biological antibacterial peptide, can be synthesized by ring-opening polymerization. However, at higher concentrations, these polymers are more hemolytic and are not suitable for systemic administration.
The hyperbranched antibacterial peptide polymer prepared by the invention has excellent inhibition effect on drug-resistant escherichia coli, and still has significant antibacterial effect at lower concentration. In addition, the hemolysis is also greatly improved, and the hemolysis rate is very low. On one hand, the production cost is reduced, on the other hand, the biocompatibility is improved, and the main obstacle is removed for the practical application of the hydrogel.
Disclosure of Invention
Aiming at the defects in the prior art, the invention aims to provide a preparation method of a hyperbranched antibacterial peptide polymer, which is reasonable in design, low in antibacterial concentration, good in sterilization effect and low in hemolysis rate, simple and convenient in synthesis method, low in cost, convenient for large-scale production and high in potential application value.
In order to achieve the purpose, the invention provides the following technical scheme:
a preparation method of hyperbranched antibacterial peptide polymer comprises the following steps:
1) and synthesis of amino-terminated hyperbranched polyamidoamine: preparing amino-terminated low molecular weight hyperbranched polyamidoamine by Michael addition reaction of methyl acrylate and diethylenetriamine;
2) lysine-N-carboxylic anhydride and valine-N-carboxylic anhydride synthesis: using hyperbranched polyamidoamine as a core, and respectively reacting benzyloxycarbonyl lysine and valine with triphosgene to obtain lysine-N-carboxylic anhydride and valine-N-carboxylic anhydride;
3) and (3) synthesizing a hyperbranched antibacterial peptide polymer: the lysine-N-carboxylic anhydride and the valine-N-carboxylic anhydride are grafted to the terminal amino group of the hyperbranched polyamidoamine through a polypeptide chain formed by ring-opening polymerization, and the antibacterial peptide polymer with the hyperbranched structure is prepared after amino deprotection and dialysis.
As an optimized technical scheme, the structure composition of the method is as follows: a core consisting of hyperbranched polymers and an external polypeptide polymer long chain.
As an optimized technical scheme, the terminal group of the hyperbranched polyamidoamine in the step 1) is amino, the number average relative molecular weight is 1000-8000, and the molecular weight distribution range is 1.2-4.0.
As an optimized technical scheme, the feeding molar ratio range of the reaction of the benzyloxycarbonyl lysine and the valine with the triphosgene in the step 2) is 1.2:1-2.0: 1; the reaction temperature is 50-60 ℃; the reaction time is 20-60 min.
As an optimized technical scheme, the polymerization mode of the lysine-N-carboxylic anhydride and the valine-N-carboxylic anhydride in the step 3) is ring-opening polymerization, and the molar ratio of the lysine-N-carboxylic anhydride to the valine-N-carboxylic anhydride in the copolymer is 1:1-5: 1.
As an optimized technical solution, the hyperbranched polyamidoamine in step 3): lysine-N-carboxylic acid anhydride: the feeding molar ratio of the valine-N-carboxylic anhydride is in the range of 1:10:5-1:100:50, wherein the ratio of the valine-N-carboxylic anhydride to the lysine-N-carboxylic anhydride is: the feed molar ratio of valine-N-carboxylic anhydride was 2: 1.
As an optimized technical scheme, the dialysis molecular weight cut-off in the step 3) is 3500Da, the dialysis time is 4d, and water is changed for many times.
As an optimized technical scheme, the relative molecular weight range of the hyperbranched antibacterial peptide polymer is 10000-100000, and the molecular weight distribution is 1.1-5.0.
By adopting the technical scheme, compared with the prior art, the hyperbranched antibacterial peptide polymer has the advantages of simple and convenient preparation method, easily obtained raw materials, lower synthesis cost and convenience for large-scale production, and meanwhile, the hyperbranched antibacterial peptide polymer prepared by the invention has good antibacterial effect and biocompatibility, and also lays a foundation for practical application and potential wide application in the fields of medicine transmission, wound treatment, medical implant materials and the like.
The invention is further illustrated with reference to the figures and examples.
Drawings
FIG. 1 is a route for the preparation of a hyperbranched antimicrobial peptide polymer according to one embodiment of the invention;
FIG. 2 is a test chart of the zone of inhibition of the hyperbranched antimicrobial peptide polymer prepared in example 1 by the Oxford cup method according to the present invention;
FIG. 3 is a comparison graph of hemolysis experiments of hyperbranched antibacterial peptide polymers prepared in the present invention.
Detailed Description
Example 1
As shown in fig. 1, a method for preparing a hyperbranched antibacterial peptide polymer comprises the following steps:
1) and (3) synthesizing amino-terminated hyperbranched polyamidoamine (H-PAMAM): dissolving a certain amount of Diethylenetriamine (DETA) in anhydrous methanol, introducing nitrogen for protection, and stirring under magnetic force. To the above solution was added dropwise a quantity of Methacrylate (MA) via syringe, wherein the molar ratio of feeds was DETA: MA ═ 1.2: 1. the reaction apparatus was then transferred to an ice-water bath, stirred for 2h and then transferred to room temperature for 48 h. And finally, transferring the system into an oil bath pot, sequentially reacting for 1h/1h/1h/1.5h at the temperature of 60 ℃/80 ℃/100 ℃/120 ℃ along with vacuumizing, and removing the solvent and unreacted monomers. After the reaction is finished, a large amount of ethyl glacial ether is used for precipitating the product for three times to obtain a yellow viscous product, namely H-PAMAM.
2) lysine-N-carboxylic anhydride (lysnca) and valine-N-carboxylic anhydride (Val NCA): the compound is obtained by respectively reacting benzyloxycarbonyl lysine and valine with triphosgene, wherein the charging molar ratio of the benzyloxycarbonyl lysine to the valine to the triphosgene is 1.5: 1. The preparation of both was similar and is illustrated by the synthesis of Lys NCA. Dispersing certain amount of benzyloxycarbonyl lysine in anhydrous tetrahydrofuran, stirring under magnetic force, and introducing nitrogen for protection. Dissolving triphosgene in a small amount of anhydrous tetrahydrofuran in a corresponding proportion, dropwise adding into the reaction system by using a syringe, heating to 50 ℃, and reacting for 0.5h until the system becomes clear. The product was precipitated using copious amounts of glacial n-hexane and recrystallized three times using n-hexane/tetrahydrofuran (v/v ═ 15:1) mixed solvent, and finally filtered off with suction to give the white product lysnca. Val NCA transparent flaky crystals were also synthesized in the same manner.
3) Synthesis of Hyperbranched Antibacterial Peptide Polymer (HAPP): H-PAMAM: lys NCA: the feeding molar ratio of Val NCA to the raw materials is 1: 20:10. Weighing a certain amount of Lys NCA and Val NCA, dissolving in anhydrous N, N-dimethylformamide, adding magnetic force for stirring, and introducing nitrogen for protection. Dissolving the H-PAMAM in a corresponding proportion in a small amount of anhydrous N, N-dimethylformamide, dropwise adding the solution into a reaction system by using a syringe, and reacting for 24 hours at room temperature. Then dropwise adding a small amount of n-butanol, continuously reacting for 1h, precipitating a product by using anhydrous ether, centrifuging and drying in vacuum. The dried product was subjected to deprotection treatment, and the product was dissolved in a mixed solution of trifluoroacetic acid (200mg/mL) and 33% HBr in glacial acetic acid (20mL/g precipitate), stirred at room temperature for 24h, and precipitated with ten-fold volume of ether. After centrifugation and drying, the product was dissolved in HCl solution (0.2M,0.2mL/mg precipitate), dialyzed against 3500Da cut-off dialysis bag for 4 days, and then lyophilized to give the final product.
Example 2
A preparation method of hyperbranched antibacterial peptide polymer comprises the following steps:
1) and (3) synthesizing amino-terminated hyperbranched polyamidoamine (H-PAMAM): dissolving a certain amount of Diethylenetriamine (DETA) in anhydrous methanol, introducing nitrogen for protection, and stirring under magnetic force. To the above solution was added dropwise a quantity of Methacrylate (MA) via syringe, wherein the molar ratio of feeds was DETA: MA ═ 1.2: 1. the reaction apparatus was then transferred to an ice-water bath, stirred for 2h and then transferred to room temperature for 48 h. And finally, transferring the system into an oil bath pot, sequentially reacting for 1h/1h/1h/1.5h/2h at 60 ℃/80 ℃/100 ℃/120 ℃/140 ℃ along with vacuumizing, and removing the solvent and unreacted monomers. After the reaction is finished, a large amount of ethyl glacial ether is used for precipitating the product for three times to obtain a yellow viscous product, namely H-PAMAM.
2) lysine-N-carboxylic anhydride (lysnca) and valine-N-carboxylic anhydride (Val NCA): the compound is obtained by respectively reacting benzyloxycarbonyl lysine and valine with triphosgene, wherein the charging molar ratio of the benzyloxycarbonyl lysine to the valine to the triphosgene is 1.5: 1. The preparation of both was similar and is illustrated by the synthesis of Lys NCA. Dispersing certain amount of benzyloxycarbonyl lysine in anhydrous tetrahydrofuran, stirring under magnetic force, and introducing nitrogen for protection. Dissolving triphosgene in a small amount of anhydrous tetrahydrofuran in a corresponding proportion, dropwise adding into the reaction system by using a syringe, heating to 50 ℃, and reacting for 0.5h until the system becomes clear. The product was precipitated using copious amounts of glacial n-hexane and recrystallized three times using n-hexane/tetrahydrofuran (v/v ═ 15:1) mixed solvent, and finally filtered off with suction to give the white product lysnca. Val NCA transparent flaky crystals were also synthesized in the same manner.
3) Synthesis of Hyperbranched Antibacterial Peptide Polymer (HAPP): H-PAMAM: lys NCA: the feeding molar ratio of Val NCA to the raw materials is 1: 20:10. Weighing a certain amount of Lys NCA and Val NCA, dissolving in anhydrous N, N-dimethylformamide, adding magnetic force for stirring, and introducing nitrogen for protection. Dissolving the H-PAMAM in a corresponding proportion in a small amount of anhydrous N, N-dimethylformamide, dropwise adding the solution into a reaction system by using a syringe, and reacting for 24 hours at room temperature. Then dropwise adding a small amount of n-butanol, continuously reacting for 1h, precipitating a product by using anhydrous ether, centrifuging and drying in vacuum. The dried product was subjected to deprotection treatment, and the product was dissolved in a mixed solution of trifluoroacetic acid (200mg/mL) and 33% HBr in glacial acetic acid (20mL/g precipitate), stirred at room temperature for 24h, and precipitated with ten-fold volume of ether. After centrifugation and drying, the product was dissolved in HCl solution (0.2M,0.2mL/mg precipitate), dialyzed against 3500Da cut-off dialysis bag for 4 days, and then lyophilized to give the final product.
Example 3
A preparation method of hyperbranched antibacterial peptide polymer comprises the following steps:
1) and (3) synthesizing amino-terminated hyperbranched polyamidoamine (H-PAMAM): dissolving a certain amount of Diethylenetriamine (DETA) in anhydrous methanol, introducing nitrogen for protection, and stirring under magnetic force. To the above solution was added dropwise a quantity of Methacrylate (MA) via syringe, wherein the molar ratio of feeds was DETA: MA ═ 1.2: 1. the reaction apparatus was then transferred to an ice-water bath, stirred for 2h and then transferred to room temperature for 48 h. And finally, transferring the system into an oil bath pot, sequentially reacting for 1h/1h/1h/1.5h at the temperature of 60 ℃/80 ℃/100 ℃/120 ℃ along with vacuumizing, and removing the solvent and unreacted monomers. After the reaction is finished, a large amount of ethyl glacial ether is used for precipitating the product for three times to obtain a yellow viscous product, namely H-PAMAM.
2) lysine-N-carboxylic anhydride (lysnca) and valine-N-carboxylic anhydride (Val NCA): the compound is obtained by respectively reacting benzyloxycarbonyl lysine and valine with triphosgene, wherein the charging molar ratio of the benzyloxycarbonyl lysine to the valine to the triphosgene is 1.5: 1. The preparation of both was similar and is illustrated by the synthesis of Lys NCA. Dispersing certain amount of benzyloxycarbonyl lysine in anhydrous tetrahydrofuran, stirring under magnetic force, and introducing nitrogen for protection. Dissolving triphosgene in a small amount of anhydrous tetrahydrofuran in a corresponding proportion, dropwise adding into the reaction system by using a syringe, heating to 50 ℃, and reacting for 0.5h until the system becomes clear. The product was precipitated using copious amounts of glacial n-hexane and recrystallized three times using n-hexane/tetrahydrofuran (v/v ═ 15:1) mixed solvent, and finally filtered off with suction to give the white product lysnca. Val NCA transparent flaky crystals were also synthesized in the same manner.
3) Synthesis of Hyperbranched Antibacterial Peptide Polymer (HAPP): H-PAMAM: lys NCA: the feeding molar ratio of Val NCA to the raw materials is 1: 50:25. Weighing a certain amount of Lys NCA and Val NCA, dissolving in anhydrous N, N-dimethylformamide, adding magnetic force for stirring, and introducing nitrogen for protection. Dissolving the H-PAMAM in a corresponding proportion in a small amount of anhydrous N, N-dimethylformamide, dropwise adding the solution into a reaction system by using a syringe, and reacting for 24 hours at room temperature. Then dropwise adding a small amount of n-butanol, continuously reacting for 1h, precipitating a product by using anhydrous ether, centrifuging and drying in vacuum. The dried product was subjected to deprotection treatment, and the product was dissolved in a mixed solution of trifluoroacetic acid (200mg/mL) and 33% HBr in glacial acetic acid (20mL/g precipitate), stirred at room temperature for 24h, and precipitated with ten-fold volume of ether. After centrifugation and drying, the product was dissolved in HCl solution (0.2M,0.2mL/mg precipitate), dialyzed against 3500Da cut-off dialysis bag for 4 days, and then lyophilized to give the final product.
Example 4
A preparation method of hyperbranched antibacterial peptide polymer comprises the following steps:
1) and (3) synthesizing amino-terminated hyperbranched polyamidoamine (H-PAMAM): dissolving a certain amount of Diethylenetriamine (DETA) in anhydrous methanol, introducing nitrogen for protection, and stirring under magnetic force. To the above solution was added dropwise a quantity of Methacrylate (MA) via syringe, wherein the molar ratio of feeds was DETA: MA ═ 1.2: 1. the reaction apparatus was then transferred to an ice-water bath, stirred for 2h and then transferred to room temperature for 48 h. And finally, transferring the system into an oil bath pot, sequentially reacting for 1h/1h/1h/1.5h/2h at 60 ℃/80 ℃/100 ℃/120 ℃/140 ℃ along with vacuumizing, and removing the solvent and unreacted monomers. After the reaction is finished, a large amount of ethyl glacial ether is used for precipitating the product for three times to obtain a yellow viscous product, namely H-PAMAM.
2) lysine-N-carboxylic anhydride (lysnca) and valine-N-carboxylic anhydride (Val NCA): the compound is obtained by respectively reacting benzyloxycarbonyl lysine and valine with triphosgene, wherein the charging molar ratio of the benzyloxycarbonyl lysine to the valine to the triphosgene is 1.5: 1. The preparation of both was similar and is illustrated by the synthesis of Lys NCA. Dispersing certain amount of benzyloxycarbonyl lysine in anhydrous tetrahydrofuran, stirring under magnetic force, and introducing nitrogen for protection. Dissolving triphosgene in a small amount of anhydrous tetrahydrofuran in a corresponding proportion, dropwise adding into the reaction system by using a syringe, heating to 50 ℃, and reacting for 0.5h until the system becomes clear. The product was precipitated using copious amounts of glacial n-hexane and recrystallized three times using n-hexane/tetrahydrofuran (v/v ═ 15:1) mixed solvent, and finally filtered off with suction to give the white product lysnca. Val NCA transparent flaky crystals were also synthesized in the same manner.
3) Synthesis of Hyperbranched Antibacterial Peptide Polymer (HAPP): H-PAMAM: lys NCA: the feeding molar ratio of Val NCA to the raw materials is 1: 50:25. Weighing a certain amount of Lys NCA and Val NCA, dissolving in anhydrous N, N-dimethylformamide, adding magnetic force for stirring, and introducing nitrogen for protection. Dissolving the H-PAMAM in a corresponding proportion in a small amount of anhydrous N, N-dimethylformamide, dropwise adding the solution into a reaction system by using a syringe, and reacting for 24 hours at room temperature. Then dropwise adding a small amount of n-butanol, continuously reacting for 1h, precipitating a product by using anhydrous ether, centrifuging and drying in vacuum. The dried product was subjected to deprotection treatment, and the product was dissolved in a mixed solution of trifluoroacetic acid (200mg/mL) and 33% HBr in glacial acetic acid (20mL/g precipitate), stirred at room temperature for 24h, and precipitated with ten-fold volume of ether. After centrifugation and drying, the product was dissolved in HCl solution (0.2M,0.2mL/mg precipitate), dialyzed against 3500Da cut-off dialysis bag for 4 days, and then lyophilized to give the final product.
Test run 1:
the hyperbranched antibacterial peptide polymer prepared in example 1 was prepared into water solutions with concentrations of 512/256/128/64/32/16/8/4/2/0 μ g/mL, and the size of the bacteriostatic region of the hyperbranched antibacterial peptide polymer on the agar medium was tested by the Oxford cup method as shown in FIG. 2. And (3) dispersedly placing the oxford cups on an agar culture medium inoculated with escherichia coli, respectively adding the hyperbranched antibacterial peptide polymer solutions with different concentrations, culturing at the constant temperature of 37 ℃ for 18 hours, and then measuring the diameter of each sample inhibition zone by using a ruler.
Test run 2:
10mg of the hyperbranched antimicrobial peptide polymer prepared in example 1 was dissolved in 10mL of physiological saline, and 0.2mL of fresh anticoagulated rabbit blood was added thereto as an experimental group. 0.2mL of fresh anticoagulated rabbit blood is added into 10mL of physiological saline to serve as a negative control group, and 0.2mL of fresh anticoagulated rabbit blood is added into 10mL of distilled water to serve as a positive control group. The sample is gently shaken evenly and then transferred to a constant temperature incubator at 37 ℃ to be kept stand for 1 h. Subsequently, the samples were centrifuged at 3000rpm for 5min, the supernatant of each sample was detected using an ultraviolet-visible spectrophotometer, and the absorbance of each sample at a wavelength of 570nm was measured, as shown in fig. 3, and the hemolysis rate was calculated according to the following formula:
wherein: ODnIs the OD value of the negative control group
ODpIs the OD value of the positive control group
ODsIs the OD value of the experimental sample group.
The protection scope of the present invention is not limited to the above embodiments, and all technical solutions belonging to the idea of the present invention belong to the protection scope of the present invention. It should be noted that modifications and embellishments within the scope of the invention may occur to those skilled in the art without departing from the principle of the invention, and are considered to be within the scope of the invention.
Claims (8)
1. A preparation method of hyperbranched antibacterial peptide polymer is characterized by comprising the following steps: the method comprises the following specific steps:
1) and synthesis of amino-terminated hyperbranched polyamidoamine: preparing amino-terminated low molecular weight hyperbranched polyamidoamine by Michael addition reaction of methyl acrylate and diethylenetriamine;
2) lysine-N-carboxylic anhydride and valine-N-carboxylic anhydride synthesis: using hyperbranched polyamidoamine as a core, and respectively reacting benzyloxycarbonyl lysine and valine with triphosgene to obtain lysine-N-carboxylic anhydride and valine-N-carboxylic anhydride;
3) and (3) synthesizing a hyperbranched antibacterial peptide polymer: the lysine-N-carboxylic anhydride and the valine-N-carboxylic anhydride are grafted to the terminal amino group of the hyperbranched polyamidoamine through a polypeptide chain formed by ring-opening polymerization, and the antibacterial peptide polymer with the hyperbranched structure is prepared after amino deprotection and dialysis.
2. The method of claim 1, wherein the hyperbranched polymer of antibacterial peptide is prepared by: the structure composition is as follows: a core consisting of hyperbranched polymers and an external polypeptide polymer long chain.
3. The method of claim 2, wherein the hyperbranched polymer of antibacterial peptide is prepared by: the terminal group of the hyperbranched polyamidoamine in the step 1) is amino, the number average relative molecular weight is 1000-8000, and the molecular weight distribution range is 1.2-4.0.
4. The method of claim 3, wherein the hyperbranched antimicrobial peptide polymer is prepared by: in the step 2), the feeding molar ratio range of the carbobenzoxy lysine and the valine reacted with the triphosgene is 1.2:1-2.0:1 respectively; the reaction temperature is 50-60 ℃; the reaction time is 20-60 min.
5. The method of claim 4, wherein the hyperbranched polymer of antibacterial peptide is prepared by: the lysine-N-carboxylic anhydride and the valine-N-carboxylic anhydride in the step 3) are polymerized by ring opening polymerization, and the molar ratio of the lysine-N-carboxylic anhydride to the valine-N-carboxylic anhydride in the copolymer is 1:1-5: 1.
6. The method of claim 5, wherein the hyperbranched polymer of antibacterial peptide is prepared by: hyperbranched polyamidoamine in step 3): lysine-N-carboxylic acid anhydride: the feeding molar ratio of the valine-N-carboxylic anhydride is in the range of 1:10:5-1:100:50, wherein the ratio of the valine-N-carboxylic anhydride to the lysine-N-carboxylic anhydride is: the feed molar ratio of valine-N-carboxylic anhydride was 2: 1.
7. The method of claim 6, wherein the hyperbranched polymer of antibacterial peptide is prepared by: in the step 3), the dialysis molecular weight cutoff is 3500Da, the dialysis time is 4d, and the water is changed for many times.
8. The method of claim 7, wherein the hyperbranched polymer of antibacterial peptide is prepared by: the relative molecular weight range of the hyperbranched antibacterial peptide polymer is 10000-100000, and the molecular weight distribution is 1.1-5.0.
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| CN110066395B (en) * | 2019-04-26 | 2020-03-10 | 山东大学 | Nano assembly based on immune checkpoint inhibitor and preparation method and application thereof |
| CN110447646B (en) * | 2019-07-11 | 2021-06-18 | 哈尔滨工程大学 | Amino acid modified branched polyetheramine cationic bacteriostatic agent and preparation method thereof |
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| CN111454457B (en) * | 2020-06-02 | 2021-03-16 | 四川大学 | Chiral peptide antibacterial polymer with dendrimer as side chain and preparation method thereof |
| CN112250884A (en) * | 2020-08-21 | 2021-01-22 | 江苏省农业科学院 | Dendritic polymeric antibacterial peptide and preparation method and application thereof |
| CN113476651A (en) * | 2021-07-12 | 2021-10-08 | 南通美韦德生命科学有限公司 | Antibacterial resin for bone fixation |
| CN114052318B (en) * | 2021-11-23 | 2024-03-08 | 深圳市北测检测技术有限公司 | Mask with drug-resistant bacteria resisting function and preparation method and application thereof |
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