CN108070045A - L- cationic chiral amino acid methacrylate copolymers and preparation and antibacterial applications - Google Patents
L- cationic chiral amino acid methacrylate copolymers and preparation and antibacterial applications Download PDFInfo
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Abstract
The present invention discloses L cationic chiral amino acid methacrylate copolymers and preparation and antibacterial applications, block copolymerization preparation is carried out with monomer L lysines methacrylate and L leucine methyls acrylate, the first L-type amino acid monomer is subjected to homopolymerization first, using 4 cyanopentanoic acid of dithiobenzoic acid as chain-transferring agent, azodiisobutyronitrile carries out RAFT polymerizations for initiator, again by L-configuration amino acid monomer polymer macromolecule chain-transferring agent, azodiisobutyronitrile is initiator, it adds in the second L-type amino acid monomer and carries out RAFT polymerizations to obtain L lysines methacrylate and L leucine methyl acrylate block copolymers.The polymer of the present invention shows excellent anti-microbial property, can significantly destroy the death that bacterium membrane structure causes bacterium；Hemolytic experiment and external smooth muscle cell compatibility experiments show that it keeps significant biocompatibility.
Description
Technical field
The invention belongs to field of biomedical polymer materials, it is related to the design synthesis side of Novel imitation antibacterial peptide polymer
Method, and in particular to a kind of preparation of cationic chiral amino acid methacrylate polymers and its antibacterial applications.
Background technology
At present, the appearance of antibiotic resistant bacteria brings great threat to publilc health health, the adherency of bacterium,
Multiplication and the biomembrane formed can cause patient to infect and a series of complication triggered even to jeopardize patient vitals.It is although alternative
The research of novel antibacterial polymer have been achieved with centainly being in progress, but it still has pole on bio-medical and clinical practice
Big challenge.Alternative novel antibacterial polymer will not only have efficient anti-microbial property, while cannot influence body group
Knit normal physiological action, in simple terms can the killing bacterium of selectivity and it is non-toxic to normal mammalian cell.
Therefore, have both high-efficiency antimicrobial activity and the antibacterial polymer of selectivity still requires study.
The content of the invention
The purpose of the present invention is preparing a kind of cationic polymethacrylate polymer based on chiral amino acid, and
Its application in antibiosis is studied, to overcome the shortcomings that existing antibacterial polymer antibacterial activity is weaker, biocompatibility is poor.
Present invention employs following technical schemes：
L-Leu methacrylate homopolymer, i.e. P (L-Leu-HEMA), with monomer D-Leu methacrylate
Homopolymerization forms, and molecular formula schematic construction is as follows：
N is the degree of polymerization, and the number-average molecular weight of polymer is 10-23kDa, molecular weight distribution index 1.10-1.25.
The preparation method of above-mentioned homopolymer forms monomer L-Leu methacrylate homopolymerization, with two thio phenyl first
Acid -4- cyanopentanoic acids are chain-transferring agent, and azodiisobutyronitrile carries out RAFT polymerizations, monomer L-Leu metering system for initiator
The molar ratio of acid esters, chain-transferring agent and initiator is (45-60)：1：0.2, preferably (50-60)：1：0.2.
In the above-mentioned technical solutions, carried out under inert gas (nitrogen, helium or argon gas) protection, reaction temperature is
60-80 degrees Celsius, when the reaction time is 6-10 small, solvent n,N-Dimethylformamide provides reaction atmosphere and environment.
In the above-mentioned technical solutions, monomer L-Leu methacrylate is prepared as steps described below：L- is bright
Propylhomoserin and hydroxyethyl methacrylate are using 4-dimethylaminopyridine as catalyst, N, N '-dicyclohexylcarbodiimide for dehydration contract
Mixture reacts 30-60 minutes by solvent of dichloromethane in ice-water bath, is stirred to react under 20-25 degrees Celsius of room temperature
When 24-48 is small, wherein L-Leu, 4-dimethylaminopyridine, N, N '-dicyclohexylcarbodiimide and hydroxyethyl methacrylate
Mass ratio be 5：(0.2—0.25)：(4—5)：(2-3), L-Leu are the L-Leu of tertbutyloxycarbonyl protection.
In the above-mentioned technical solutions, the homopolymer of preparation is deprotected under trifluoroacetic acid effect, preferably by homopolymer
It is dissolved in dichloromethane, under trifluoroacetic acid effect, when progress deprotection reaction 2-4 is small under 20-25 degrees Celsius, after purification
Dissolving dialysis, freeze-drying.
Due to being acted on using trifluoroacetic acid, with trifluoroacetic acid group in the polymer after deprotection reaction, due to Leu's
Isoelectric point is pH 6.01, and the isoelectric point of Lys is pH 9.60, when adjusting pH is 7.2 ,-the NH of leu3 +Generation deprotonation, it is raw
Into-NH2, shown in following chemical formula：
D-Leu methacrylate homopolymer, i.e. P (D-Leu-HEMA), with monomer D-Leu methacrylate
Homopolymerization forms, and molecular formula schematic construction is as follows, and n is the degree of polymerization, and the number-average molecular weight of polymer is 10-23kDa, molecular weight distribution
Coefficient is 1.10-1.25.
The preparation method of above-mentioned homopolymer forms monomer D-Leu methacrylate homopolymerization, with two thio phenyl first
Acid -4- cyanopentanoic acids are chain-transferring agent, and azodiisobutyronitrile carries out RAFT polymerizations, monomer D-Leu metering system for initiator
The molar ratio of acid esters, chain-transferring agent and initiator is (45-60)：1：0.2, preferably (50-60)：1：0.2.
In the above-mentioned technical solutions, carried out under inert gas (nitrogen, helium or argon gas) protection, reaction temperature is
60-80 degrees Celsius, when the reaction time is 6-10 small, solvent n,N-Dimethylformamide provides reaction atmosphere and environment.
In the above-mentioned technical solutions, monomer D-Leu methacrylate is prepared as steps described below：D- is bright
Propylhomoserin and hydroxyethyl methacrylate are using 4-dimethylaminopyridine as catalyst, N, N '-dicyclohexylcarbodiimide for dehydration contract
Mixture reacts 30-60 minutes by solvent of dichloromethane in ice-water bath, is stirred to react under 20-25 degrees Celsius of room temperature
When 24-48 is small, wherein D-Leu, 4-dimethylaminopyridine, N, N '-dicyclohexylcarbodiimide and hydroxyethyl methacrylate
Mass ratio be 5：(0.2—0.25)：(4—5)：(2-3), D-Leu are the D-Leu of tertbutyloxycarbonyl protection.
In the above-mentioned technical solutions, the homopolymer of preparation is deprotected under trifluoroacetic acid effect, preferably by homopolymer
It is dissolved in dichloromethane, under trifluoroacetic acid effect, when progress deprotection reaction 2-4 is small under 20-25 degrees Celsius, after purification
Dissolving dialysis, freeze-drying.
Due to being acted on using trifluoroacetic acid, with trifluoroacetic acid group in the polymer after deprotection reaction, due to Leu's
Isoelectric point is pH 6.01, and the isoelectric point of Lys is pH 9.60, when adjusting pH is 7.2 ,-the NH of leu3 +Generation deprotonation, it is raw
Into-NH2, shown in following chemical formula：
L-lysine methacrylate homopolymer, i.e. P (L-Lys-HEMA), with monomer L-lysine methacrylate
Homopolymerization forms, and molecular formula schematic construction is as follows, and n is the degree of polymerization, and the number-average molecular weight of polymer is 10-23kDa, molecular weight distribution
Coefficient is 1.10-1.25.
The preparation method of above-mentioned homopolymer forms monomer L-lysine methacrylate homopolymerization, with two thio phenyl first
Acid -4- cyanopentanoic acids are chain-transferring agent, and azodiisobutyronitrile carries out RAFT polymerizations, monomer L-lysine metering system for initiator
The molar ratio of acid esters, chain-transferring agent and initiator is (45-60)：1：0.2, preferably (50-60)：1：0.2.
In the above-mentioned technical solutions, carried out under inert gas (nitrogen, helium or argon gas) protection, reaction temperature is
60-80 degrees Celsius, when the reaction time is 6-10 small, solvent n,N-Dimethylformamide provides reaction atmosphere and environment.
In the above-mentioned technical solutions, monomer L-lysine methacrylate is prepared as steps described below：L- is relied
Propylhomoserin and hydroxyethyl methacrylate are using 4-dimethylaminopyridine as catalyst, N, N '-dicyclohexylcarbodiimide for dehydration contract
Mixture reacts 30-60 minutes by solvent of dichloromethane in ice-water bath, is stirred to react under 20-25 degrees Celsius of room temperature
When 24-48 is small, wherein L-lysine, 4-dimethylaminopyridine, N, N '-dicyclohexylcarbodiimide and hydroxyethyl methacrylate
Mass ratio be 5：(0.2—0.25)：(4—5)：(2-3), L-lysine are the L-lysine of tertbutyloxycarbonyl protection.
In the above-mentioned technical solutions, the homopolymer of preparation is deprotected under trifluoroacetic acid effect, preferably by homopolymer
It is dissolved in dichloromethane, under trifluoroacetic acid effect, when progress deprotection reaction 2-4 is small under 20-25 degrees Celsius, after purification
Dissolving dialysis, freeze-drying.
Due to being acted on using trifluoroacetic acid, with trifluoroacetic acid group in the polymer after deprotection reaction, adjusting pH is
7.2, lys is unaffected, shown in following chemical formula：
D-Lys methacrylate homopolymer, i.e. P (D-Leu-HEMA), with monomer D-Lys methacrylate
Homopolymerization forms, and molecular formula schematic construction is as follows, and n is the degree of polymerization, and the number-average molecular weight of polymer is 10-23kDa, molecular weight distribution
Coefficient is 1.10-1.25.
The preparation method of above-mentioned homopolymer forms monomer D-Lys methacrylate homopolymerization, with two thio phenyl first
Acid -4- cyanopentanoic acids are chain-transferring agent, and azodiisobutyronitrile carries out RAFT polymerizations, monomer D-Lys metering system for initiator
The molar ratio of acid esters, chain-transferring agent and initiator is (45-60)：1：0.2, preferably (50-60)：1：0.2.
In the above-mentioned technical solutions, carried out under inert gas (nitrogen, helium or argon gas) protection, reaction temperature is
60-80 degrees Celsius, when the reaction time is 6-10 small, solvent n,N-Dimethylformamide provides reaction atmosphere and environment.
In the above-mentioned technical solutions, monomer D-Lys methacrylate is prepared as steps described below：D- is relied
Propylhomoserin and hydroxyethyl methacrylate are using 4-dimethylaminopyridine as catalyst, N, N '-dicyclohexylcarbodiimide for dehydration contract
Mixture reacts 30-60 minutes by solvent of dichloromethane in ice-water bath, is stirred to react under 20-25 degrees Celsius of room temperature
When 24-48 is small, wherein D-Lys, 4-dimethylaminopyridine, N, N '-dicyclohexylcarbodiimide and hydroxyethyl methacrylate
Mass ratio be 5：(0.2—0.25)：(4—5)：(2-3), D-Lys are the D-Lys of tertbutyloxycarbonyl protection.
In the above-mentioned technical solutions, the homopolymer of preparation is deprotected under trifluoroacetic acid effect, preferably by homopolymer
It is dissolved in dichloromethane, under trifluoroacetic acid effect, when progress deprotection reaction 2-4 is small under 20-25 degrees Celsius, after purification
Dissolving dialysis, freeze-drying.
Due to being acted on using trifluoroacetic acid, with trifluoroacetic acid group in the polymer after deprotection reaction, adjusting pH is
7.2, lys is unaffected, shown in following chemical formula：
D- cationic chiral amino acid methacrylate copolymers, it is bright with monomer D-Lys methacrylate and D-
Propylhomoserin methacrylate carry out block copolymerization preparation, molecular formula schematic construction is as follows, n, m be respective monomer the degree of polymerization, m/n
For (0.8-1.2), the consistent m=n of the degree of polymerization of preferably two kinds monomers, the number-average molecular weight of polymer is 10-23kDa, molecular weight
Breadth coefficient is 1.10-1.25.
The preparation method of above-mentioned copolymer, is prepared as steps described below：
First D types amino acid monomer is carried out homopolymerization by step 1., using dithiobenzoic acid -4- cyanopentanoic acids as chain tra nsfer
Agent, azodiisobutyronitrile carry out RAFT polymerizations to obtain D amino acids monomer-polymers, the first D type ammonia for initiator
The molar ratio of base acid monomers, chain-transferring agent and initiator is (45-60)：1：0.2, preferably (50-60)：1：0.2；
Step 2, the D amino acids monomer-polymer Macromolecular chain transfer agents that prepared by step 1, azodiisobutyronitrile are to draw
Agent is sent out, the 2nd D types amino acid monomer is added in and carries out RAFT polymerizations to obtain D-Lys methacrylate and D-Leu first
Base acrylate block copolymer, the molar ratio of the 2nd D types amino acid monomer, Macromolecular chain transfer agent and initiator are
(80—100)：1：0.2, preferably (85-90)：1：0.2.
In the above-mentioned technical solutions, in step 1, carried out under inert gas (nitrogen, helium or argon gas) protection, instead
Answering temperature, when the reaction time is 6-10 small, solvent n,N-Dimethylformamide provides reaction atmosphere and ring for 60-80 degrees Celsius
Border.
In the above-mentioned technical solutions, in step 2, carried out under inert gas (nitrogen, helium or argon gas) protection, instead
Answering temperature, when the reaction time is 6-10 small, solvent n,N-Dimethylformamide provides reaction atmosphere and ring for 60-80 degrees Celsius
Border.
In the above-mentioned technical solutions, monomer D-Lys methacrylate is prepared as steps described below：D- is relied
Propylhomoserin and hydroxyethyl methacrylate are using 4-dimethylaminopyridine as catalyst, N, N '-dicyclohexylcarbodiimide for dehydration contract
Mixture reacts 30-60 minutes by solvent of dichloromethane in ice-water bath, is stirred to react under 20-25 degrees Celsius of room temperature
When 24-48 is small, wherein D-Lys, 4-dimethylaminopyridine, N, N '-dicyclohexylcarbodiimide and hydroxyethyl methacrylate
Mass ratio be 5：(0.2—0.25)：(4—5)：(2-3), D-Lys are the D-Lys of tertbutyloxycarbonyl protection.
In the above-mentioned technical solutions, monomer D-Leu methacrylate is prepared as steps described below：D- is bright
Propylhomoserin and hydroxyethyl methacrylate are using 4-dimethylaminopyridine as catalyst, N, N '-dicyclohexylcarbodiimide for dehydration contract
Mixture reacts 30-60 minutes by solvent of dichloromethane in ice-water bath, is stirred to react under 20-25 degrees Celsius of room temperature
When 24-48 is small, wherein D-Leu, 4-dimethylaminopyridine, N, N '-dicyclohexylcarbodiimide and hydroxyethyl methacrylate
Mass ratio be 5：(0.2—0.25)：(4—5)：(2-3), D-Leu are the D-Leu of tertbutyloxycarbonyl protection.
In the above-mentioned technical solutions, the copolymer of preparation is deprotected under trifluoroacetic acid effect, preferably by copolymer
It is dissolved in dichloromethane, under trifluoroacetic acid effect, when progress deprotection reaction 2-4 is small under 20-25 degrees Celsius, after purification
Dissolving dialysis, freeze-drying.
Due to being acted on using trifluoroacetic acid, with trifluoroacetic acid group in the polymer after deprotection reaction, due to Leu's
Isoelectric point is pH 6.01, and the isoelectric point of Lys is pH 9.60, when adjusting pH is 7.2 ,-the NH of leu3 +Generation deprotonation, it is raw
Into-NH2, it is that 7.2, lys is unaffected to adjust pH, shown in following chemical formula：
L- cationic chiral amino acid methacrylate copolymers, it is bright with monomer L-lysine methacrylate and L-
Propylhomoserin methacrylate carry out block copolymerization preparation, molecular formula schematic construction is as follows, n, m be respective monomer the degree of polymerization, m/n
For (0.8-1.2), the consistent m=n of the degree of polymerization of preferably two kinds monomers, the number-average molecular weight of polymer is 10-23kDa, molecular weight
Breadth coefficient is 1.10-1.25.
The preparation method of above-mentioned copolymer, is prepared as steps described below：
First L-type amino acid monomer is carried out homopolymerization by step 1., using dithiobenzoic acid -4- cyanopentanoic acids as chain tra nsfer
Agent, azodiisobutyronitrile carry out RAFT polymerizations to obtain D amino acids monomer-polymers, the first L-type ammonia for initiator
The molar ratio of base acid monomers, chain-transferring agent and initiator is (45-60)：1：0.2, preferably (50-60)：1：0.2；
Step 2, the L-configuration amino acid monomer polymer macromolecule chain-transferring agent that prepared by step 1, azodiisobutyronitrile are to draw
Agent is sent out, the second L-type amino acid monomer is added in and carries out RAFT polymerizations to obtain L-lysine methacrylate and L-Leu first
Base acrylate block copolymer, the molar ratio of the second L-type amino acid monomer, Macromolecular chain transfer agent and initiator are
(80—100)：1：0.2, preferably (85-90)：1：0.2.
In the above-mentioned technical solutions, in step 1, carried out under inert gas (nitrogen, helium or argon gas) protection, instead
Answering temperature, when the reaction time is 6-10 small, solvent n,N-Dimethylformamide provides reaction atmosphere and ring for 60-80 degrees Celsius
Border.
In the above-mentioned technical solutions, in step 2, carried out under inert gas (nitrogen, helium or argon gas) protection, instead
Answering temperature, when the reaction time is 6-10 small, solvent n,N-Dimethylformamide provides reaction atmosphere and ring for 60-80 degrees Celsius
Border.
In the above-mentioned technical solutions, monomer L-lysine methacrylate is prepared as steps described below：L- is relied
Propylhomoserin and hydroxyethyl methacrylate are using 4-dimethylaminopyridine as catalyst, N, N '-dicyclohexylcarbodiimide for dehydration contract
Mixture reacts 30-60 minutes by solvent of dichloromethane in ice-water bath, is stirred to react under 20-25 degrees Celsius of room temperature
When 24-48 is small, wherein L-lysine, 4-dimethylaminopyridine, N, N '-dicyclohexylcarbodiimide and hydroxyethyl methacrylate
Mass ratio be 5：(0.2—0.25)：(4—5)：(2-3), L-lysine are the L-lysine of tertbutyloxycarbonyl protection.
In the above-mentioned technical solutions, monomer L-Leu methacrylate is prepared as steps described below：L- is bright
Propylhomoserin and hydroxyethyl methacrylate are using 4-dimethylaminopyridine as catalyst, N, N '-dicyclohexylcarbodiimide for dehydration contract
Mixture reacts 30-60 minutes by solvent of dichloromethane in ice-water bath, is stirred to react under 20-25 degrees Celsius of room temperature
When 24-48 is small, wherein L-Leu, 4-dimethylaminopyridine, N, N '-dicyclohexylcarbodiimide and hydroxyethyl methacrylate
Mass ratio be 5：(0.2—0.25)：(4—5)：(2-3), L-Leu are the L-Leu of tertbutyloxycarbonyl protection.
In the above-mentioned technical solutions, the copolymer of preparation is deprotected under trifluoroacetic acid effect, preferably by copolymer
It is dissolved in dichloromethane, under trifluoroacetic acid effect, when progress deprotection reaction 2-4 is small under 20-25 degrees Celsius, after purification
Dissolving dialysis, freeze-drying.
Due to being acted on using trifluoroacetic acid, with trifluoroacetic acid group in the polymer after deprotection reaction, due to Leu's
Isoelectric point is pH 6.01, and the isoelectric point of Lys is pH 9.60, when adjusting pH is 7.2 ,-the NH of leu3 +Generation deprotonation, it is raw
Into-NH2, it is that 7.2, lys is unaffected to adjust pH, shown in following chemical formula：
The present invention provides a kind of method of regulation and control copolymer overall configuration, i.e., is total to using the monomer of D configurations and L-configuration
It is poly-, by the ingredient proportion of D types/L-type monomer, molar ratio between the two is changed, so that copolymer shows as D types or L-type,
Application of the i.e. controllable living polymerization in leucine-lysine methacrylate copolymer chirality is regulated and controled.
Using reversible addion-fragmentation chain transfer polymerization by the ingredient proportion of D types/L-type monomer, change is between the two
Molar ratio so that copolymer shows as D types or L-type, i.e., by D types and the ingredient proportion of L-type monomer, change in copolymer
Molar ratio between D types monomer and L-type monomer, so that copolymer general performance is D types or L-type.
For above-mentioned four kinds of monomers, polymer that homopolymerization obtains shows the D and L-configuration consistent with corresponding monomer, and two
The copolymer that a L-type monomer copolymerization obtains shows as L-type, and the copolymer that two D type monomer copolymerizations obtain shows as D types, copolymerization
Object is copolymerized using D types and L-type monomer, adjusts the relative populations of D types monomer and L-type monomer, you can realizes that copolymer is whole
Configuration shows controllable.
In the technical solution of the present invention, the control of (i.e. monomer molar ratio) of adding materials can be realized using living polymerization,
Reaction condition is mild, and synthon yield is 70%-85%, polymerization conversion 50%-90%, homopolymer and copolymer
Molecular weight it is controllable and breadth coefficient is relatively narrow.
The invention discloses based on chiral amino acid methacrylate polymers and preparation method thereof and antibacterial applications.Circle
Dichroscope spectrum characterization result proves that above-mentioned obtained L-configuration and D configurations polymer have opposite deuterostrophies structure.Microdilution
Antibacterial experiment research show the dialysis of above-mentioned gained it is lyophilized after the methacrylate polymers based on chiral amino acid minimum suppression
Bacteria concentration (MIC) is 100~1000 μ gmL-1, excellent anti-microbial property is shown, field emission scanning electron microscope result figure is intuitively
The death that bacterium membrane structure causes bacterium can significantly be destroyed by showing antibacterial polymer produced by the present invention；Hemolytic experiment and
External smooth muscle cell compatibility experiments show cationic chiral amino acid methacrylate antimicrobial polymerizable prepared by the present invention
Object can still keep significant biocompatibility under concentration needed for antibacterial, and wherein hemolysis rate is less than 10%, and smooth muscle cell is lived
Property can reach more than 80%.
Description of the drawings
Fig. 1 is the mechanism of polymerization schematic diagram of reversible addion-fragmentation chain transfer polymerization (RAFT) in the present invention.
Fig. 2 be in the present invention monomer D-Leu (Boc)-HEMA in CDCl3In1H-NMR spectrum.
Fig. 3 be in the present invention monomer D-Lys (Boc)-HEMA in CDCl3In1H-NMR spectrum.
Fig. 4 is P (D-Leu (Boc)-HEMA) homopolymer (1) not being deprotected in the present invention, P (D-Lys (Boc)-HEMA)
Homopolymer (2) and P (D-Leu (Boc)-HEMA)-b-P (D-Lys (Boc)-HEMA) block copolymer (3) are in CDCl3In1H-
NMR spectra.
Fig. 5 be in the present invention be deprotected after P (D-Leu-HEMA) homopolymer (1), P (D-Lys-HEMA) homopolymer homopolymer
(2) and P (D-Leu-HEMA)-b-P (D-Lys-HEMA) block copolymer homopolymers (3) are in D2In O1H-NMR spectrum.
Fig. 6 is the circular dichroism spectra characterization result figure of homopolymer and copolymer in the present invention.
Fig. 7 is the stereoscan photograph of polymer antibacterial experiment in the present invention.
Fig. 8 is the antibacterial experiment effect curve figure (1) that polymer is directed to Escherichia coli in the present invention.
Fig. 9 is the antibacterial experiment effect curve figure (1) that polymer is directed to staphylococcus aureus in the present invention.
Figure 10 is the biocompatibility in vitro phenogram of Inventive polymers P (L-Leu-HEMA).
Figure 11 is the biocompatibility in vitro phenogram of Inventive polymers P (D-Leu-HEMA).
Figure 12 is the biocompatibility in vitro phenogram of Inventive polymers P (D-Leu-HEMA-b-D-Lys-HEMA).
Figure 13 is the biocompatibility in vitro phenogram of Inventive polymers P (L-Leu-HEMA-b-L-Lys-HEMA).
Specific embodiment
The technical solution further illustrated the present invention with reference to specific embodiment.The experimental raw and instrument that embodiment uses
Device difference is as shown in the table：
(1) experimental raw
(2) laboratory apparatus
Polymerize in embodiments of the present invention using reversible addion-fragmentation chain transfer, RAFT polymerizations be a kind of activity/controllable from
It is polymerize by base, is applicable in the monomer containing double bond functional group.In RAFT polymerizations, conventional initiator is thermally decomposed into primary group of free radicals
I., and trigger monomer polymerization into Propagating Radical Pn ., the C=S keys in Propagating Radical and chain-transferring agent carry out reversible addition shape
Into intermediate dormancy kind, S-R keys are broken in dormancy kind, form new reactive species free radical Rn ., then trigger monomer polymerization, react machine
Reason is illustrated in fig. 1 shown below.Different from conventional free radical polymerization, RAFT polymerization chain tra nsfers are reversible process, intermediate dormancy kind and life
The balanced reaction of reversible addition and reversible fracture is carried out between long-chain free radical, so that it is guaranteed that all chains are given birth to equal probability
It is long, form the polymer of narrow ditribution, number of free radical maintains a relative constant reduced levels in system, in inhibition system
The biradical termination reaction of free radical so that polymerization activity is controllable.Initiator is used in the present invention and triggers the first monomer, with the
One monomer and chain-transferring agent carry out living polymerization, and when adding in second comonomer, supplement initiator is to trigger second comonomer, with activity
It polymerize the living polymerization that the first obtained monomer homopolymers carry out second comonomer for Macromolecular chain transfer agent.
Embodiment 1-prepare monomer D-Leu hydroxyethyl methacrylate, L-Leu hydroxyethyl methacrylate, D-
Lysine hydroxyethyl methacrylate and L-lysine hydroxyethyl methacrylate
The chiral amino acid monomer 5g for being dissolved in 22mL dry methylene chlorides is added in first into dry neck round bottom flask
(it is respectively D-Leu, L-Leu, D-Lys and L-lysine, to ensure the activity of functional group in reaction, selects tertiary fourth
The boc-protected above-mentioned four kinds of amino acid of oxygen carbonyl, sloughs Boc protecting groups using trifluoroacetic acid after reacting), in magnetic agitation
Logical nitrogen purification down, then adds in the catalyst DMAP 0.24g for being dissolved in 1.5mL dry methylene chlorides, reaction bulb is placed in ice water
It in bath, is slowly added to be dissolved in the dehydrating condensation agent DCC 4.53g of 20mL dry methylene chlorides dropwise, and exists under nitrogen protection
2.86g HEMA are added in 20min.30min is reacted under ice-water bath, under subsequent nitrogen atmosphere under 20-25 degrees Celsius of room temperature
The reaction was continued 36h.It filters after reaction and removes white precipitate, obtained organic mixed solution 70mL distilled water and 100mL
Dichloromethane extracts four times, and obtained organic layer is further extracted twice with 60mL0.1mol/L hydrochloric acid solutions successively, and 60mL satisfies
It is washed twice with sodium bicarbonate solution and 60mL saturated nacl aqueous solutions.It is finally dried, is stirred overnight with anhydrous sodium sulfate, filtered
Afterwards, revolving removes solvent.
Four kinds of monomers of preparation are characterized using nuclear magnetic resonance, as a result as shown in Figures 2 and 3, in view of four kinds of monomers
Chemical composition be divided into two kinds (i.e. leucine methyl hydroxy-ethyl acrylates and lysine hydroxyethyl methacrylate), chemical structure
It is divided into D and L, the nuclear magnetic spectrogram of the various configuration of same chemical composition is basically identical, is understood as shown in two attached drawings, nuclear-magnetism is common
The chemical shift shaken corresponds to the hydrogen atom of the different chemical environments marked in chemical formula, exactly proves that four kinds of monomers are successfully prepared.
Embodiment 2-and using four kinds of monomers of the preparation of embodiment 1 as raw material, reversible addion-fragmentation chain transfer is used to polymerize and is made
Standby four kinds of homopolymers
Cationic chiral amino acid hydroxyethyl methacrylate homopolymer is prepared using RAFT polymerizations：With magnetic force
In Schlenk bottles of the 25mL of stirrer, add in monomer made from 1.5g embodiments 1, CPADB 24.4mg, AIBN 2.86mg and
1.5g anhydrous DMF solvents-are taken out-by freezing three times after melting the foreign gas that Xun Huan is removed in reaction system, are placed in 70 DEG C of oil bath
In pot, when reaction 10 is small under nitrogen protection.After the completion of reaction, exposure is placed in being quickly cooled down termination in ice-water bath in air
Reaction, then with acetone/n-hexane repeated precipitation 5 times.Obtained product is placed in 30 DEG C of vacuum drying chamber dry 8h, is obtained
The sample of each homopolymer.Under the conditions of ice-water bath, the ratio of 20mL dichloromethane and 10mL trifluoroacetic acids is added according to every 1g samples
Example, is added in reaction bulb, then in room temperature reaction 3h, rotates to remove solvent.Products therefrom is added dropwise to anhydrous ether, instead
Multiple precipitation is three times.Obtained product is placed in 30 DEG C of vacuum drying chamber dry 8h.Product after purification is dissolved in deionization
In water, dialysis 72h removes remaining impurity, and pure spongy solid sample is obtained after freeze-drying.
Four kinds of homopolymers of preparation are characterized using nuclear magnetic resonance, as a result as shown in figs. 4 and 5, similarly based on bright
There are the difference (D of chemical structure for the two kinds of polymer of propylhomoserin hydroxyethyl methacrylate and lysine hydroxyethyl methacrylate
Type and L-type), the nuclear magnetic spectrogram of the various configuration of same chemical composition is basically identical, is understood as shown in two attached drawings, nuclear-magnetism is common
The chemical shift shaken corresponds to the hydrogen atom of the different chemical environments marked in chemical formula, exactly proves that four kinds of homopolymers are successfully made
It is standby, and block polymer does not generate variation after deprotection.
Embodiment 3-preparation D- cationic chiral amino acid hydroxyethyl methacrylate copolymers
(1) homopolymer of D-leucine is prepared first using the method for embodiment 2：In the 25mL with magnetic stir bar
In Schlenk bottles, add in monomer D-leucine methyl hydroxy-ethyl acrylate prepared by 1.5g embodiments 1, CPADB 24.4mg,
AIBN 2.86mg and 1.5g anhydrous DMF solvents-are taken out-by freezing three times after melting the foreign gas that Xun Huan is removed in reaction system,
It is placed in 70 DEG C of oil bath pan, when reaction 10 is small under nitrogen protection, after treating that reaction is basic, without the end of exposure air
Only react, and what the active group (CPADB) for being to maintain D-leucine homopolymer end was reacted as addition second comonomer
Macromolecular chain transfer agent；
(2) in reaction vessel, lysine methacrylate monomers (the i.e. D-lysine first of 3g identical configurations is added in
Base hydroxy-ethyl acrylate), AIBN 2.15mg and with 3g anhydrous DMF solvents, with the Macromolecular chain transfer agent 0.96g prepared before
It is reacted ,-takes out-after melting the foreign gas that Xun Huan is removed in reaction system, be placed in 70 DEG C of oil bath pan by freezing three times,
When the lower reaction 6 of nitrogen protection is small, obtained product is placed in 30 DEG C of vacuum drying chamber dry 8h after multiple deposition and purification；
(3) under the conditions of ice-water bath, 20mL dichloromethane is added according to every 1g samples (sample that i.e. above-mentioned steps 2 obtain)
It with the ratio of 10mL trifluoroacetic acids, is added in reaction bulb, then in room temperature reaction 3h, rotates to remove solvent.Gained is produced
Object is added dropwise to anhydrous ether, and repeated precipitation is three times.Obtained product is placed in 30 DEG C of vacuum drying chamber dry 8h.It will purifying
Product afterwards is dissolved in deionized water, and dialysis 72h removes remaining impurity, and pure spongy solid sample is obtained after freeze-drying
Product.Obtain the cationic chiral amino acid hydroxyethyl methacrylate copolymer of D configurations.
The cationic chiral amino acid hydroxyethyl methacrylate copolymer of the D configurations of preparation is carried out using nuclear magnetic resonance
Characterization, as shown in figs. 4 and 5, the chemical shift of nuclear magnetic resonance corresponds to the hydrogen atom of the different chemical environments marked in chemical formula,
Exactly prove that copolymer is successfully prepared, and block polymer does not generate variation after deprotection.
Embodiment 4-preparation L- cationic chiral amino acid methacrylate copolymers
(1) homopolymer of L-leucine is prepared first using the method for embodiment 2：In the 25mL with magnetic stir bar
In Schlenk bottles, add in monomer L-leucine methyl hydroxy-ethyl acrylate prepared by 1.5g embodiments 1, CPADB 24.4mg,
AIBN 2.86mg and 1.5g anhydrous DMF solvents-are taken out-by freezing three times after melting the foreign gas that Xun Huan is removed in reaction system,
It is placed in 70 DEG C of oil bath pan, when reaction 10 is small under nitrogen protection, after treating that reaction is basic, without the end of exposure air
Only react, and what the active group (CPADB) for being to maintain L-leucine homopolymer end was reacted as addition second comonomer
Macromolecular chain transfer agent；
(2) in reaction vessel, lysine methacrylate monomers (the i.e. L-lysine first of 3g identical configurations is added in
Base hydroxy-ethyl acrylate), AIBN 2.15mg and with 3g anhydrous DMF solvents, with the Macromolecular chain transfer agent 0.96g prepared before
It is reacted ,-takes out-after melting the foreign gas that Xun Huan is removed in reaction system, be placed in 70 DEG C of oil bath pan by freezing three times,
When the lower reaction 6 of nitrogen protection is small, obtained product is placed in 30 DEG C of vacuum drying chamber dry 8h after multiple deposition and purification；
(3) under the conditions of ice-water bath, 20mL dichloromethane is added according to every 1g samples (sample that i.e. above-mentioned steps 2 obtain)
It with the ratio of 10mL trifluoroacetic acids, is added in reaction bulb, then in room temperature reaction 3h, rotates to remove solvent.Gained is produced
Object is added dropwise to anhydrous ether, and repeated precipitation is three times.Obtained product is placed in 30 DEG C of vacuum drying chamber dry 8h.It will purifying
Product afterwards is dissolved in deionized water, and dialysis 72h removes remaining impurity, and pure spongy solid sample is obtained after freeze-drying
Product.Obtain the cationic chiral amino acid hydroxyethyl methacrylate copolymer of L-configuration.
The copolymer of preparation is characterized using nuclear magnetic resonance, the cationic chiral ammonia of the D configurations prepared with embodiment 3
Base acids methacrylic acid hydroxyl ethyl ester copolymer is compared, and only there is different (L-types), the chemical environment basic one of hydrogen atom in configuration
It causes, i.e. the chemical shift of D types and L-type is basically identical, and as a result as shown in figs. 4 and 5, the chemical shift of nuclear magnetic resonance corresponds to chemistry
The hydrogen atom of the different chemical environments marked in formula exactly proves that copolymer is successfully prepared, and the bulk polymerization after deprotection
Object does not generate variation.
Embodiment 5-prepare D-Lys/L-Leu methacrylate copolymer
(1) homopolymer of L-leucine is prepared first using the method for embodiment 2：In the 25mL with magnetic stir bar
In Schlenk bottles, add in monomer L-leucine methyl hydroxy-ethyl acrylate prepared by 1.5g embodiments 1, CPADB 24.4mg,
AIBN 2.86mg and 1.5g anhydrous DMF solvents-are taken out-by freezing three times after melting the foreign gas that Xun Huan is removed in reaction system,
It is placed in 70 DEG C of oil bath pan, when reaction 10 is small under nitrogen protection, after treating that reaction is basic, without the end of exposure air
Only react, and what the active group (CPADB) for being to maintain L-leucine homopolymer end was reacted as addition second comonomer
Macromolecular chain transfer agent；
(2) in reaction vessel, lysine methacrylate monomers (the i.e. D-lysine first of 3g various configurations is added in
Base hydroxy-ethyl acrylate), AIBN 2.15mg and with 3g anhydrous DMF solvents, with the Macromolecular chain transfer agent 0.96g prepared before
It is reacted ,-takes out-after melting the foreign gas that Xun Huan is removed in reaction system, be placed in 70 DEG C of oil bath pan by freezing three times,
When the lower reaction 6 of nitrogen protection is small, obtained product is placed in 30 DEG C of vacuum drying chamber dry 8h after multiple deposition and purification；
(3) under the conditions of ice-water bath, 20mL dichloromethane is added according to every 1g samples (sample that i.e. above-mentioned steps 2 obtain)
It with the ratio of 10mL trifluoroacetic acids, is added in reaction bulb, then in room temperature reaction 3h, rotates to remove solvent.Gained is produced
Object is added dropwise to anhydrous ether, and repeated precipitation is three times.Obtained product is placed in 30 DEG C of vacuum drying chamber dry 8h.It will purifying
Product afterwards is dissolved in deionized water, and dialysis 72h removes remaining impurity, and pure spongy solid sample is obtained after freeze-drying
Product obtain D-Lys/L-Leu methacrylate copolymer.
Embodiment 6-prepare D-Leu/L-lysine methacrylate copolymer
(1) homopolymer of D-leucine is prepared first using the method for embodiment 2：In the 25mL with magnetic stir bar
In Schlenk bottles, add in monomer D-leucine methyl hydroxy-ethyl acrylate prepared by 1.5g embodiments 1, CPADB 24.4mg,
AIBN 2.86mg and 1.5g anhydrous DMF solvents-are taken out-by freezing three times after melting the foreign gas that Xun Huan is removed in reaction system,
It is placed in 70 DEG C of oil bath pan, when reaction 10 is small under nitrogen protection, after treating that reaction is basic, without the end of exposure air
Only react, and what the active group (CPADB) for being to maintain D-leucine homopolymer end was reacted as addition second comonomer
Macromolecular chain transfer agent；
(2) in reaction vessel, lysine methacrylate monomers (the i.e. L-lysine first of 3g various configurations is added in
Base hydroxy-ethyl acrylate), AIBN 2.15mg and with 3g anhydrous DMF solvents, with the Macromolecular chain transfer agent 0.96g prepared before
It is reacted ,-takes out-after melting the foreign gas that Xun Huan is removed in reaction system, be placed in 70 DEG C of oil bath pan by freezing three times,
When the lower reaction 6 of nitrogen protection is small, obtained product is placed in 30 DEG C of vacuum drying chamber dry 8h after multiple deposition and purification；
(3) under the conditions of ice-water bath, 20mL dichloromethane is added according to every 1g samples (sample that i.e. above-mentioned steps 2 obtain)
It with the ratio of 10mL trifluoroacetic acids, is added in reaction bulb, then in room temperature reaction 3h, rotates to remove solvent.Gained is produced
Object is added dropwise to anhydrous ether, and repeated precipitation is three times.Obtained product is placed in 30 DEG C of vacuum drying chamber dry 8h.It will purifying
Product afterwards is dissolved in deionized water, and dialysis 72h removes remaining impurity, and pure spongy solid sample is obtained after freeze-drying
Product.Obtain D-Leu/L-lysine methacrylate copolymer.
The polymer obtained by four kinds of monomer homopolymerizations shows the D and L-configuration consistent with corresponding monomer, two L-type monomers
It is copolymerized obtained copolymer and shows as L-type, the copolymer that two D type monomer copolymerizations obtain shows as D types, prepared by embodiment 5 and 6
Copolymer be copolymerized using D types and L-type monomer, can rubbing between the two be changed by the ingredient proportion of D types/L-type monomer
That ratio, so that copolymer shows as D types or L-type, i.e., passes through D types and L-type using reversible addion-fragmentation chain transfer polymerization
The ingredient proportion of monomer changes the molar ratio between D types monomer and L-type monomer in copolymer, so that copolymer general performance is D
Type or L-type.
7-homopolymer of embodiment, the test of copolymer property
As shown in fig. 6, the D configurations of preparation and L-configuration polymer are subjected to circular dichroism spectra test, in identical wave-length coverage
Interior appearance upward wave crest and downward trough, the corresponding polymer of upward wave crest is L-configuration, to the corresponding polymer of lower wave trough
For D configurations, antipodal helical conformation is shown.
Microdilution antibacterial experiment bibliography：P.Li,C.C.Zhou,S.Rayatpisheh,K.Ye,Y.F.Poon,
P.T.Hammond,H.W.Duan,M.B.Chan-Park,Adv.Mater.,2012,24,4130-4137；External human aorta
Smooth muscle cell compatibility experiments bibliography：S.E.Exley,L.C.Paslay,G.S.Sahukhal,B.A.Abel,
T.D.Brown,C.L.McCormick,S.Heinhorst,V.Koul,V.Choudhary,M.O.Elasri,S.E.Morgan,
Biomacromolecules,2015,16,3845-3852；Hemolytic experiment bibliography：A.Pascual,J.P.K.Tan,
A.Yuen,J.M.W.Chan,D.J.Coady,D.Mecerreyes,J.L.Hedrick,Y.Y.Yang,H.Sardon,
Biomacromolecules,2015,16,1169-1178。
Staphylococcus aureus is selected to be represented for gram-positive bacteria, Escherichia coli represent for Gram-negative bacteria, first
Bacterium bacterial strain is incubated overnight and reaches growth medium, every milliliter 3 × 10 is diluted to PBS buffer solution8A Colony Forming Unit
(CFU), Polymer Solution (aqueous solution of the polymer of preparation) aseptic liquid nutrient medium doubling dilution is secondly taken into 100 μ L
The bacterial suspension diluted is added in 96 orifice plates in the Polymer Solution of 100 μ L, and the ultimate density for obtaining antibacterial polymer is
2-4096μg·mL-1.96 orifice plates are then placed in 37 DEG C of culture 18h, use absorbance of the microplate reader test microvia at 600nm
Value.Using bacterial cell culture solutions of the 200 μ L without antibacterial polymer as positive controls, culture solution pure 200 μ L is feminine gender
Control group.Experiment is respectively provided with 4-6 parallel sample every time.Simultaneously a series of control group of concentration polymer solutions is set to exclude
Influence of the polymer solution to experimental group absorbance.It then, will with the glutaraldehyde phosphate buffer of 2.5% percentage by volume
Bacterium (20-25 degrees Celsius) the fixed 4h at room temperature of polymer solution culture contact 3h by 100 μ L lethal doses, and
With PBS buffer solution rinsing twice, 15min is then dehydrated successively by the ethanol solution of gradient concentration, is centrifuged under 1000rpm
10min, it was evenly coated on silicon chip, natural air drying, metal spraying 50s observes bacterium under field emission scanning electron microscope
Surface breakdown pattern.
As shown in Figure 7, antibacterial polymer produced by the present invention is intuitively shown from field emission scanning electron microscope result figure
The death that bacterium membrane structure causes bacterium can be significantly destroyed, and the polymer according to above-described embodiment preparation is shown and base
This consistent performance.As shown in figs. 8 and 9, P (D-Leu-HEMA) homopolymer, P (D-Lys-HEMA) homopolymers and P (D-Leu-
HEMA)-b-P (D-Lys-HEMA) block copolymer shows excellent anti-microbial property, the experimental results showed that, P (D-Leu-
HEMA) minimum inhibitory concentration (MIC) of homopolymer is 798~820 μ gmL-1, the MIC of P (D-Lys-HEMA) homopolymer is 100
~120 μ gmL-1, the MIC of P (D-Leu-HEMA)-b-P (D-Lys-HEMA) block copolymer is 180~200 μ gmL-1
(showing basically identical situation for Escherichia coli and staphylococcus aureus)；Use the homopolymer based on L-type amino acid instead
And copolymer, show the homopolymer and copolymer with the basically identical anti-microbial property of above-described embodiment, slightly below D type amino acid.
The copolymer being copolymerized using D types and L-type monomer, by adjusting the ingredient proportion of D types/L-type monomer, copolymer shows as D
Type or L-type equally have the anti-microbial property consistent with respective configuration, less than the anti-microbial property of respective configuration.
As shown in attached drawing 10-13, hemolytic experiment and external human aortic smooth muscle cell's compatibility experiments show this hair
The polymer of bright preparation can still keep significant biocompatibility under concentration needed for antibacterial, and wherein hemolysis rate is less than 10%,
Smooth muscle cell activity can reach more than 80%.
The preparation of above-mentioned polymer is carried out according to present invention, polymer shows the property basically identical with embodiment
Energy.Illustrative description is done to the present invention above, it should which explanation in the case where not departing from the core of the present invention, is appointed
What simple deformation, modification or other skilled in the art can not spend the equivalent substitution of creative work to each fall within this
The protection domain of invention.
Claims (10)
1.L- cationic chiral amino acid methacrylate copolymers, which is characterized in that with monomer L-lysine metering system
Acid esters and L-Leu methacrylate carry out block copolymerization preparation, and molecular formula schematic construction is as follows, n, and m is respective monomer
The degree of polymerization.
2. L- cationic chirals amino acid methacrylate copolymer according to claim 1, which is characterized in that m/n
For (0.8-1.2), the consistent m=n of the degree of polymerization of preferably two kinds monomers.
3. L- cationic chirals amino acid methacrylate copolymer according to claim 1, which is characterized in that polymerization
The number-average molecular weight of object is 10-23kDa, molecular weight distribution index 1.10-1.25.
The preparation method of 4.L- cationic chiral amino acid methacrylate copolymers, which is characterized in that as steps described below
It is prepared：
First L-type amino acid monomer is carried out homopolymerization by step 1., even using dithiobenzoic acid -4- cyanopentanoic acids as chain-transferring agent
Nitrogen bis-isobutyronitrile carries out RAFT polymerizations to obtain D amino acids monomer-polymers, the first L-type amino acid list for initiator
The molar ratio of body, chain-transferring agent and initiator is (45-60)：1：0.2；
Step 2, the L-configuration amino acid monomer polymer macromolecule chain-transferring agent that prepared by step 1, azodiisobutyronitrile are initiation
Agent adds in the second L-type amino acid monomer and carries out RAFT polymerizations to obtain L-lysine methacrylate and L-Leu methyl
Acrylate block copolymer, the molar ratio of the second L-type amino acid monomer, Macromolecular chain transfer agent and initiator are
(80—100)：1：0.2.
5. the preparation method of L- cationic chirals amino acid methacrylate copolymer according to claim 4, special
Sign is, in step 1, is carried out under inert gas (nitrogen, helium or argon gas) protection, reaction temperature is Celsius for 60-80
Degree, when the reaction time is 6-10 small, solvent n,N-Dimethylformamide provides reaction atmosphere and environment；First L-type amino acid list
The molar ratio of body, chain-transferring agent and initiator is (50-60)：1：0.2.
6. the preparation method of L- cationic chirals amino acid methacrylate copolymer according to claim 4, special
Sign is, in step 2, is carried out under inert gas (nitrogen, helium or argon gas) protection, reaction temperature is Celsius for 60-80
Degree, when the reaction time is 6-10 small, solvent n,N-Dimethylformamide provides reaction atmosphere and environment；Second L-type amino acid list
The molar ratio of body, Macromolecular chain transfer agent and initiator is (85-90)：1：0.2.
7. the preparation method of L- cationic chirals amino acid methacrylate copolymer according to claim 4, special
Sign is that monomer L-lysine methacrylate is prepared as steps described below：By L-lysine and hydroxyethyl methacrylate
Ethyl ester is using 4-dimethylaminopyridine as catalyst, N, N '-dicyclohexylcarbodiimide be dehydrating condensation agent, using dichloromethane to be molten
Agent is reacted 30-60 minutes in ice-water bath, be stirred to react under 20-25 degrees Celsius of room temperature 24-48 it is small when, wherein L- relies ammonia
The mass ratio of acid, 4-dimethylaminopyridine, N, N '-dicyclohexylcarbodiimide and hydroxyethyl methacrylate is 5：(0.2—
0.25)：(4—5)：(2-3), L-lysine are the L-lysine of tertbutyloxycarbonyl protection；Monomer L-Leu methacrylic acid
Ester is prepared as steps described below：By L-Leu and hydroxyethyl methacrylate using 4-dimethylaminopyridine as catalyst,
N, N '-dicyclohexylcarbodiimide be dehydrating condensation agent, reacted by solvent of dichloromethane in ice-water bath 30-60 minutes, then
Be stirred to react under 20-25 degrees Celsius of room temperature 24-48 it is small when, wherein L-Leu, 4-dimethylaminopyridine, N ,-two hexamethylenes of N '
The mass ratio of base carbodiimide and hydroxyethyl methacrylate is 5：(0.2—0.25)：(4—5)：(2-3), L-Leu are
The L-Leu of tertbutyloxycarbonyl protection.
8. the preparation method of L- cationic chirals amino acid methacrylate copolymer according to claim 4, special
Sign is that the copolymer of preparation is deprotected under trifluoroacetic acid effect, preferably copolymer is dissolved in dichloromethane, three
Under fluoroacetic acid effect, when progress deprotection reaction 2-4 is small under 20-25 degrees Celsius, dissolving dialysis, freeze-drying after purification is
It can.
9. the L- cationic chiral amino acid methacrylate copolymers as described in claims 1 or 2 or 3 are in antibacterial
Application, which is characterized in that 100~1000 μ gmL of minimum inhibitory concentration average out to-1, polymer can destroy bacterium membrane structure
Cause the death of bacterium, and keep significant biocompatibility.
10. application according to claim 9, which is characterized in that bacterium is staphylococcus aureus or Escherichia coli.
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| CN103193926A (en) * | 2013-04-18 | 2013-07-10 | 苏州大学 | Copolymer containing lysine residue on side chain and preparation method thereof as well as fibrinolytic functional material |
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| CN102775530A (en) * | 2012-05-30 | 2012-11-14 | 江南大学 | RAFT (reversible addition fragmentation chain transfer) preparation method of polylysine derivative |
| CN103193926A (en) * | 2013-04-18 | 2013-07-10 | 苏州大学 | Copolymer containing lysine residue on side chain and preparation method thereof as well as fibrinolytic functional material |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113925060A (en) * | 2021-09-13 | 2022-01-14 | 浙江理工大学 | Nano ZnO loaded antibacterial polymer and preparation method and application thereof |
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