CN110330656A - Amphipathic graft copolymer and its preparation method and application containing antibacterial peptide - Google Patents
Amphipathic graft copolymer and its preparation method and application containing antibacterial peptide Download PDFInfo
- Publication number
- CN110330656A CN110330656A CN201910608080.1A CN201910608080A CN110330656A CN 110330656 A CN110330656 A CN 110330656A CN 201910608080 A CN201910608080 A CN 201910608080A CN 110330656 A CN110330656 A CN 110330656A
- Authority
- CN
- China
- Prior art keywords
- antibacterial peptide
- graft copolymer
- hydrophobic polymer
- caprolactone
- amphipathic graft
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/74—Synthetic polymeric materials
- A61K31/785—Polymers containing nitrogen
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/34—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
- A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
- A61K47/60—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G63/00—Macromolecular compounds obtained by reactions forming a carboxylic ester link in the main chain of the macromolecule
- C08G63/02—Polyesters derived from hydroxycarboxylic acids or from polycarboxylic acids and polyhydroxy compounds
- C08G63/06—Polyesters derived from hydroxycarboxylic acids or from polycarboxylic acids and polyhydroxy compounds derived from hydroxycarboxylic acids
- C08G63/08—Lactones or lactides
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G63/00—Macromolecular compounds obtained by reactions forming a carboxylic ester link in the main chain of the macromolecule
- C08G63/78—Preparation processes
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G63/00—Macromolecular compounds obtained by reactions forming a carboxylic ester link in the main chain of the macromolecule
- C08G63/91—Polymers modified by chemical after-treatment
- C08G63/912—Polymers modified by chemical after-treatment derived from hydroxycarboxylic acids
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G81/00—Macromolecular compounds obtained by interreacting polymers in the absence of monomers, e.g. block polymers
Abstract
The present invention provides a kind of amphipathic graft copolymer and its preparation method and application containing antibacterial peptide, the amphipathic graft copolymer is connected with antibacterial peptide by covalent bond by the segment of hydrophobic polymer to be made, as antibacterial agent or pharmaceutical carrier in terms of clinical anti-infective, the anticancer drug Targeting delivery or nanosecond medical science in applied；Amphipathic graft copolymer of the invention has selected the good hydrophobic polymer segment of biocompatibility instead of the hydrophobic amino acid segment in antibacterial peptide, therefore does not have cytotoxicity, and has excellent biocompatibility and biodegradability；In addition, it is with excellent broad spectrum antibacterial performance, it is identical that Antibacterial Mechanism is destroyed with the similar film of natural antibacterial peptide, Induction of bacterial is not easy and generates drug resistance；Structure-controllable, the synthesis step of amphipathic graft copolymer containing antibacterial peptide of the invention are easy, low in cost, and can be realized industrialization production.
Description
Technical field
The invention belongs to medical macromolecular materials technical fields, and in particular to a kind of amphipathic graft copolymerization containing antibacterial peptide
Object and its preparation method and application.
Background technique
From the discovery of penicillin, antibiotic is on antibacterial therapy it may be said that bringing Gospel to the mankind.However in recent years by
In antibiotic abuse and abuse, lead to the generation of drug-fast bacteria and superbacteria, therefore find and be different from conventional antibiotic antibacterial
The excellent antibacterial agent of mechanism is extremely urgent.Natural antibacterial peptide is found in the intracorporal immune system of biology, is not only had wide
Efficient sterilizing activity is composed, and the film failure mechanism of antibacterial peptide is different from conventional antibiotic, i.e., positively charged antibacterial peptide passes through quiet
Electro ultrafiltration is adsorbed onto the surface of negatively charged bacterial cell membrane, and then hydrophobic polymeric segment is inserted into bacterial cell membrane,
Cause to generate hole on bacterial cell membrane, bacterium content outflow, to kill bacterium.This special film destroys Antibacterial Mechanism
Bacterium is set to be difficult to generate drug resistance to antibacterial peptide.However the extraction process of natural antibacterial peptide is complicated, low output and at high cost, because
This chemically synthesized mode, which synthesizes antibacterial agent similar with natural antibacterial peptide structure, far-reaching research significance.At the same time, portion
Dividing the hydrophobic amino acid residues such as phenyl ring etc. of antibacterial peptide has larger toxicity to human normal cell, constrains antibacterial peptide in clinic
The development in the fields such as medical treatment, synthesis no cytotoxicity, good biocompatibility and degradable antibacterial agent are always that anti-biotic material is ground
The common objective for the person of studying carefully.
Summary of the invention
In view of the deficiencies of the prior art, primary and foremost purpose of the invention is to provide a kind of amphipathic graft copolymerization containing antibacterial peptide
Object.
Second object of the present invention is to provide a kind of preparation method of amphipathic graft copolymer containing antibacterial peptide.
Third object of the present invention is to provide a kind of application of amphipathic graft copolymer containing antibacterial peptide.
In order to achieve the above objectives, solution of the invention is:
A kind of amphipathic graft copolymer containing antibacterial peptide, structural formula are as follows:
Wherein, integer of the J in 1-50, integer of the K in 1-50, integer of the L in 1-50.J, K and L difference
The degree of polymerization of random copolymerization block in graft copolymer is represented, Pep indicates antibacterial peptide.
Preferably, antibacterial peptide is selected from one or more of melittin, epsilon-polylysine, LL-37, bacitracin and cecropin.
Preferably, hydrophobic polymer is poly- (6-caprolactone) of the carboxyl functional group of second containing side.
A kind of preparation method of the above-mentioned amphipathic graft copolymer containing antibacterial peptide comprising following steps:
(1), with methoxy poly (ethylene glycol) (mPEG) for macromole evocating agent, stannous iso caprylate is catalyst, by ε-in oneself
The 6-caprolactone monomer 6- (acetic acid benzyl ester base -6-caprolactone) (BCL) of ester and function dough ring opening copolymer in organic solvent
Synthesize 6-caprolactone/6- (acetic acid benzyl ester base)-epsilon-caprolactone copolymer (PCL-co- containing side acetic acid benzyl ester base
PBCL benzyl protection group), and under palladium carbon catalytic hydrogenation conditions is sloughed, a series of the poly- of main chain carboxylic groups containing second has been obtained
(6-caprolactone) (mPEG-b-P (2-CCL-co-6-CCL)), as hydrophobic polymer, are denoted as G, structural formula are as follows:
Wherein, J and K respectively represents the degree of polymerization of random copolymerization block in hydrophobic polymer (G)；
(2), by hydrophobic polymer, N, N '-dicyclohexylcarbodiimide and n-hydroxysuccinimide are in organic solvent
Interior reaction obtains the hydrophobic polymer segment that carboxyl is activated, structural formula are as follows:
Wherein, J and K respectively represents the degree of polymerization of random copolymerization block in hydrophobic polymer (G)；
(3), the hydrophobic polymer segment that carboxyl is activated is dissolved in organic solvent, and antibacterial peptide is added and carries out instead
It answers, after washed, precipitating, drying, obtains the amphipathic graft copolymer containing antibacterial peptide, structural formula are as follows:
Wherein, integer of the J in 1-50, integer of the K in 1-50, integer of the L in 1-50.J, K and L difference
The degree of polymerization of random copolymerization block in graft copolymer is represented, Pep indicates antibacterial peptide.
Preferably, antibacterial peptide is selected from one or more of melittin, epsilon-polylysine, LL-37, bacitracin and cecropin.
Preferably, hydrophobic polymer is poly- (6-caprolactone) of the carboxyl functional group of second containing side.
Preferably, in step (1), step (2) and step (3), organic solvent is selected from tetrahydrofuran, N, N- dimethyl formyl
Any one or a few in amine, methylene chloride, toluene, p-methyl benzenesulfonic acid, morpholine, acetone and dimethyl sulfoxide.
Preferably, in step (2), the molar ratio of hydrophobic polymer segment and antibacterial peptide that carboxyl is activated is 1:(1-
100)。
Preferably, the temperature of the reaction of step (2) is -10-60 DEG C, and the time of reaction is 1-24h.
Preferably, the temperature of the reaction of step (3) is -10-60 DEG C, and the time of reaction is 1-24h.
A kind of above-mentioned amphipathic graft copolymer containing antibacterial peptide as antibacterial agent or pharmaceutical carrier it is clinical it is anti-infective,
It is applied in terms of anticancer drug Targeting delivery or nanosecond medical science.
By adopting the above scheme, the beneficial effects of the present invention are:
The first, the amphipathic graft copolymer of the invention containing antibacterial peptide has selected the good hydrophobicity of biocompatibility high
Molecule segment does not have cytotoxicity instead of the hydrophobic amino acid segment in antibacterial peptide, and has excellent biofacies
Capacitive and biodegradability.
The second, the amphipathic graft copolymer of the invention containing antibacterial peptide has excellent broad spectrum antibacterial performance, antibacterial
Mechanism is identical as the film of natural antibacterial peptide destruction Antibacterial Mechanism, is not easy Induction of bacterial and generates drug resistance.
Third, the structure-controllable of the amphipathic graft copolymer of the invention containing antibacterial peptide, synthesis step are easy, at low cost
It is honest and clean, and can be realized industrialization production.
Detailed description of the invention
Fig. 1 is the structural formula of the amphipathic graft copolymer of the invention containing antibacterial peptide.
Fig. 2 is the schematic diagram of the amphipathic graft copolymer of the invention containing antibacterial peptide.
Specific embodiment
The present invention provides a kind of amphipathic graft copolymer and its preparation method and application containing antibacterial peptide.
<amphipathic graft copolymer containing antibacterial peptide>
As depicted in figs. 1 and 2, the amphipathic graft copolymer of the invention containing antibacterial peptide, by hydrophobic polymer
Segment, which is connected with antibacterial peptide by covalent bond, to be made, and structural formula is as follows:
Wherein, integer of the J in 1-50, integer of the K in 1-50, integer of the L in 1-50.J, K and L difference
The degree of polymerization of random copolymerization block in graft copolymer is represented, Pep indicates natural or artificial synthesized antibacterial peptide.
(antibacterial peptide)
Antibacterial peptide can be selected from melittin (melittin), epsilon-polylysine (EPL), LL-37, bacitracin and cecropin etc.
Natural or artificial synthesized one or more of antibacterial peptide.
(hydrophobic polymer)
Hydrophobic polymer is poly- (6-caprolactone) of the carboxyl functional group of second containing side.
In fact, the above-mentioned amphipathic graft copolymer containing antibacterial peptide is similar to the structure of natural cationic type antibacterial peptide,
Therefore its Antibacterial Mechanism is also similar to natural cationic type antibacterial peptide, is film damage type Antibacterial Mechanism.Above-mentioned amphipathic grafting is altogether
Positively charged antibacterial peptide generates electrostatic adsorption with the bacterial cell membrane with negative electrical charge on polymers branch, to be adsorbed onto
On bacterium, then inside hydrophobic polymer segment insertion cell membrane, the integrality of film is destroyed, is caused intracellular in cell membrane
It is tolerant to leak, finally cause bacterial death.Therefore, when above-mentioned amphipathic graft copolymer is used as antibacterial agent, it is thin to be also not easy induction
Bacterium generates drug resistance.
<preparation method of the amphipathic graft copolymer containing antibacterial peptide>
The preparation method of amphipathic graft copolymer containing antibacterial peptide of the invention includes the following steps:
(1), with methoxy poly (ethylene glycol) (mPEG) for macromole evocating agent, stannous iso caprylate is catalyst, by ε-in oneself
The 6-caprolactone monomer 6- (acetic acid benzyl ester base -6-caprolactone) (BCL) of ester and function dough ring opening copolymer in organic solvent
Synthesize 6-caprolactone/6- (acetic acid benzyl ester base)-epsilon-caprolactone copolymer (PCL-co- containing side acetic acid benzyl ester base
PBCL benzyl protection group), and under palladium carbon catalytic hydrogenation conditions is sloughed, a series of the poly- of main chain carboxylic groups containing second has been obtained
(ε-caprolactone) (mPEG-b-P (2-CCL-co-6-CCL)), as hydrophobic polymer, are denoted as G, structural formula are as follows:
Wherein, J and K respectively represents the degree of polymerization of random copolymerization block in hydrophobic polymer (G)；
(2), by hydrophobic polymer, N, N '-dicyclohexylcarbodiimide and n-hydroxysuccinimide are in organic solvent
Interior reaction obtains the hydrophobic polymer segment that carboxyl is activated, structural formula are as follows:
Wherein, J and K respectively represents the degree of polymerization of random copolymerization block in hydrophobic polymer (G)；
(3), the hydrophobic polymer segment that carboxyl is activated is dissolved in organic solvent, and antibacterial peptide is added and carries out instead
It answers, after washed, precipitating, drying, obtains the amphipathic graft copolymer containing antibacterial peptide, structural formula are as follows:
Wherein, integer of the J in 1-50, integer of the K in 1-50, integer of the L in 1-50.J, K and L difference
The degree of polymerization of random copolymerization block in graft copolymer is represented, Pep indicates antibacterial peptide.
(antibacterial peptide)
Antibacterial peptide is selected from natural or artificial synthesized anti-such as melittin, epsilon-polylysine, LL-37, bacitracin and cecropin
One or more of bacterium peptide.
(hydrophobic polymer)
Hydrophobic polymer is poly- (6-caprolactone) of the carboxyl functional group of second containing side.
(organic solvent)
In step (1), step (2) and step (3), organic solvent is selected from tetrahydrofuran (THF), N, N- dimethyl formyl
It is any in amine (DMF), methylene chloride (DCM), toluene, p-methyl benzenesulfonic acid, morpholine, acetone and dimethyl sulfoxide (DMSO)
It is one or more of.
(hydrophobic polymer segment and antibacterial peptide that carboxyl is activated)
In step (2), the molar ratio of hydrophobic polymer segment and antibacterial peptide that carboxyl is activated can be 1:(1-
100), preferably 1:100, more preferably 1:10.
In step (2), the temperature of reaction can be -10-60 DEG C, preferably 25 DEG C；The time of reaction can be 1-
For 24 hours, preferably 8h.
In step (3), the temperature of reaction can be -10-60 DEG C, preferably 25 DEG C；The time of reaction can be 1-
For 24 hours, preferably 8h.
<purposes of the amphipathic graft copolymer containing antibacterial peptide>
The above-mentioned amphipathic graft copolymer containing antibacterial peptide can be used as antibacterial agent in clinical anti-infective, anticancer drug target
It is applied to release or nanosecond medical science etc..
The above-mentioned amphipathic graft copolymer containing antibacterial peptide can also be used as pharmaceutical carrier in clinical anti-infective, anticarcinogen
Object Targeting delivery or nanosecond medical science etc. are applied.
In fact, it is the poly- of the obtained main chain carboxylic group containing second of primary raw material that the present invention, which is selected with poly- (6-caprolactone),
(6-caprolactone) block copolymer and natural or artificial synthesized antibacterial peptide pass through reaction and above-mentioned two containing antibacterial peptide are prepared
Parent's property graft copolymer.Wherein, hydrophilic antimicrobial peptide segment has a large amount of positive charges, with negatively charged bacterial phospholipid bilayer
Electrostatic interaction is generated to be adsorbed on cell membrane, while in hydrophobic polymer segment insertion phospholipid bilayer, destroying cell
The integrality of film is to achieve the purpose that bactericidal, since poly- (6-caprolactone) is that biocompatibility is good, biodegradable
Property it is excellent and there is the macromolecule through release property to drug, than containing the isostructural hydrophobic amino acid segment toxicity of phenyl ring
It is low.Therefore, the above-mentioned amphipathic graft copolymer no cytotoxicity containing antibacterial peptide prepared by the present invention, biocompatibility it is good and
Biodegradability is good, as antibacterial agent or pharmaceutical carrier in clinical anti-infective, anticancer drug Targeting delivery or nanosecond medical science
Etc. be all with a wide range of applications.
The present invention is further illustrated with reference to embodiments.
Embodiment 1:
The preparation method of the amphipathic graft copolymer G-EPL containing epsilon-polylysine of the present embodiment includes the following steps:
(1) using methoxy poly (ethylene glycol) (mPEG) as initiator, stannous iso caprylate catalysis under, by 6-caprolactone with
6-caprolactone monomer 6- (acetic acid benzyl ester base -6-caprolactone) (BCL) ring-opening copolymerization in tetrahydrofuran of 10g function dough
At 6-caprolactone/6- (acetic acid benzyl ester base)-epsilon-caprolactone copolymer (PCL-co-PBCL) containing side acetic acid benzyl ester base.
Then the benzyl protecting group that PCL-co-PBCL is sloughed by palladium/carbon catalytic hydrogenation finally obtains the poly- of the carboxyl functional group of second containing side
(6-caprolactone) (PCL-co-PCCL), as hydrophobic polymer, are denoted as G, structural formula are as follows:
Wherein, J and K respectively represents the degree of polymerization of random copolymerization block in hydrophobic polymer (G)；
(2) the hydrophobic polymer G of 200mg main chain carboxyl containing second is dissolved in 10mL tetrahydrofuran, is placed in ice-water bath
In.It is dissolved in 5mL tetrahydrofuran, 5mg DCC and 3mg HOSu (1.1 times that take the amount of DCC substance) then in 30min
It is instilled in the tetrahydrofuran solution of hydrophobic polymer of main chain carboxyl group dropwise.Ice-water bath is removed, is stirred at 25 DEG C of room temperature
Mix reaction 8h.After reaction by product washing, precipitating, drying, the hydrophobic polymer segment that carboxyl is activated is obtained, is tied
Structure formula are as follows:
Wherein, J and K respectively represents the degree of polymerization of random copolymerization block in hydrophobic polymer；
(3) 1.0g epsilon-polylysine (EPL) is added in the hydrophobic polymer solution that above-mentioned 200mg carboxyl is activated,
Being added in appropriate tetrahydrofuran dissolves it sufficiently, and regulation system pH value, is stirred to react 8h at 25 DEG C.It revolves after reaction
Turn evaporation of solvent, acetone be added and simultaneously stirs removing impurity, be then collected by filtration solid, evaporation remove be added after acetone go from
Sub- water, stirring dissolve it sufficiently.Dialyse 48h in deionized water, and during which every 2h changes a water, then rotates and steams at 60 DEG C
Hair removes water, and the amphipathic graft copolymer G-EPL containing epsilon-polylysine, structural formula can be obtained are as follows:
Wherein, integer of the J in 1-50, integer of the K in 1-50, integer of the L in 1-50.J, K and L difference
Represent the degree of polymerization of random copolymerization block in graft copolymer.
In fact, the temperature of reaction is within -10-60 DEG C, the time of reaction is within 1-24h in step (2)
Can with.
In step (3), the temperature of reaction is within -10-60 DEG C, the time of reaction is possible within 1-24h.
Embodiment 2:
The preparation method of the amphipathic graft copolymer G-melittin containing melittin of the present embodiment includes the following steps:
(1) with methoxy poly (ethylene glycol) (mPEG) for initiator, under stannous iso caprylate catalysis, by 6-caprolactone and 10g
The 6-caprolactone monomer 6- (acetic acid benzyl ester base -6-caprolactone) (BCL) of function dough ring opening copolymer in tetrahydrofuran synthesizes
6-caprolactone/6- (acetic acid benzyl ester base)-epsilon-caprolactone copolymer (PCL-co-PBCL) containing side acetic acid benzyl ester base.So
The benzyl protecting group for taking off PCL-co-PBCL by palladium/carbon catalytic hydrogenation afterwards, obtaining the carboxyl functional group of second containing side poly-, (ε-is in oneself
Ester) (PCL-co-PCCL), as hydrophobic polymer, it is denoted as G, structural formula are as follows:
Wherein, J and K respectively represents the degree of polymerization of random copolymerization block in hydrophobic polymer (G)；
(2) hydrophobic polymer of 200mg main chain carboxyl containing second is dissolved in 10mL tetrahydrofuran, is placed in ice-water bath
In.It is dissolved in 5mL tetrahydrofuran, 5mg DCC and 3mg HOSu (1.1 times that take the amount of DCC substance) then in 30min
It is instilled in the tetrahydrofuran solution of hydrophobic polymer of main chain carboxyl group dropwise.Ice-water bath is removed, is stirred at 25 DEG C of room temperature
Mix reaction 8h.After reaction by product washing, precipitating, drying, the hydrophobic polymer segment that carboxyl is activated is obtained, is tied
Structure formula are as follows:
Wherein, J and K respectively represents the degree of polymerization of random copolymerization block in hydrophobic polymer；
(3) the hydrophobic polymer solution that 200mg carboxyl is activated among the above is added in 1.0g melittin (melittin)
In, being added in appropriate tetrahydrofuran dissolves it sufficiently, and regulation system pH value, is stirred to react 8h at 25 DEG C.After reaction
Rotary evaporation removes solvent, and acetone is added and stirs removing impurity, and solid is then collected by filtration, is added and goes after evaporation removing acetone
Ionized water, stirring dissolve it sufficiently.Dialyse 48h in deionized water, and during which every 2h changes a water, then rotates at 60 DEG C
Evaporation removes water, and the amphipathic graft copolymer G-melittin containing epsilon-polylysine, structural formula can be obtained are as follows:
Wherein, integer of the J in 1-50, integer of the K in 1-50, integer of the L in 1-50.J, K and L difference
Represent the degree of polymerization of random copolymerization block in graft copolymer.
Embodiment 3:
The preparation method of the amphipathic graft copolymer G- (LL-37) containing LL-37 of the present embodiment includes the following steps:
(1) with methoxy poly (ethylene glycol) (mPEG) for initiator, under stannous iso caprylate catalysis, by 6-caprolactone and 10g
The 6-caprolactone monomer 6- (acetic acid benzyl ester base -6-caprolactone) (BCL) of function dough ring opening copolymer in tetrahydrofuran synthesizes
6-caprolactone/6- (acetic acid benzyl ester base)-epsilon-caprolactone copolymer (PCL-co-PBCL) containing side acetic acid benzyl ester base.So
The benzyl protecting group for taking off PCL-co-PBCL by palladium/carbon catalytic hydrogenation afterwards, obtaining the carboxyl functional group of second containing side poly-, (ε-is in oneself
Ester) (PCL-co-PCCL), as hydrophobic polymer, it is denoted as G, structural formula are as follows:
Wherein, J and K respectively represents the degree of polymerization of random copolymerization block in hydrophobic polymer；
(2) hydrophobic polymer of 200mg main chain carboxyl containing second is dissolved in 10mL tetrahydrofuran, is placed in ice-water bath
In.It is dissolved in 5mL tetrahydrofuran, 5mg DCC and 3mg HOSu (1.1 times that take the amount of DCC substance) then in 30min
It is instilled in the tetrahydrofuran solution of hydrophobic polymer of main chain carboxyl group dropwise.Ice-water bath is removed, is stirred at 25 DEG C of room temperature
Mix reaction 8h.After reaction by product washing, precipitating, drying, the hydrophobic polymer segment that carboxyl is activated is obtained, is tied
Structure formula are as follows:
Wherein, J and K respectively represents the degree of polymerization of random copolymerization block in hydrophobic polymer；
(3) 1.0g LL-37 is added in the hydrophobic polymer solution that 200mg carboxyl is activated among the above, is added appropriate
It is dissolved sufficiently in tetrahydrofuran, and regulation system pH value, be stirred to react 8h at 25 DEG C.Rotary evaporation removes after reaction
Solvent is added acetone and stirs removing impurity, and solid is then collected by filtration, and deionized water is added after removing acetone in evaporation, stirs
Dissolve it sufficiently.Dialyse 48h in deionized water, and during which every 2h changes a water, and then rotary evaporation removes water at 60 DEG C,
The amphipathic graft copolymer G- (LL-37) containing epsilon-polylysine, structural formula can be obtained are as follows:
Wherein, integer of the J in 1-50, integer of the K in 1-50, integer of the L in 1-50.J, K and L difference
Represent the degree of polymerization of random copolymerization block in graft copolymer.
<experiment>
Distinguished using the good amphipathic graft copolymer containing antibacterial peptide of the biocompatibility of above-described embodiment as product
It is tested as follows.
This experiment is to verify good amphipathic graft copolymer (G-EPL, the G- containing antibacterial peptide of biocompatibility
Melittin and G- (LL-37)) in gram-positive bacteria (staphylococcus aureus) and Gram-negative bacteria (Escherichia coli)
Anti-microbial property.
Wherein, minimal inhibitory concentration (MIC) is to assess the important parameter of antibacterial antiplaque agent performance.This experiment is respectively using leather
Lan Shi negative bacterium (Escherichia coli) and gram-positive bacteria (staphylococcus aureus) are good containing anti-to measure biocompatibility
The anti-microbial property of the amphipathic graft copolymer of bacterium peptide.
Experimental procedure is as follows:
(1) the activated Escherichia coli of 1mL and staphylococcus aureus are taken respectively, remove LB meat bone with physiological saline
Soup, and the strain of the strain of Escherichia coli and staphylococcus aureus is separately added into and is uniformly mixed in 1mL physiological saline, it is
Facilitate bacterium colony when coated plate to count, bacterium liquid decimal dilution method is diluted 105Times；
(2) configuration concentration is the amphipathic graft copolymer solution of 1000 μ g/mL, obtains various concentration with double of dilution method
Amphipathic graft copolymer solution, concentration is followed successively by 1000 μ g/mL, 500 μ g/mL, 250 μ g/mL, 125 μ g/mL, 64 μ g/
mL,32μg/mL,16μg/m,8μg/mL,4μg/mL；100 μ L are respectively taken to be placed in 1mL centrifuge tube；
(3) bacterium liquid (Escherichia coli or golden yellow after 100 μ L dilution is added in each centrifuge tube in step (2)
Staphylococcus), amphipathic graft copolymer concentration become 500 μ g/mL, 250 μ g/mL, 125 μ g/mL, 64 μ g/mL, 32 μ g/mL,
16 μ g/mL, 8 μ g/mL, 4 μ g/mL, 2 μ g/mL, cultivate 2h under 37 DEG C of constant temperature, and amphiphilic is not added for similarity condition culture
The pure bacterium of property graft copolymer is as control sample；
(4) mixed liquor of 100 μ L various concentrations is added in culture dish, the 10mL agar solution of warm is added, rocks
Culture dish is buckled to agar solidification after even, is placed under 37 DEG C of constant temperatures after cultivating 18h and carries out bacterium colony counting, observation inhibits thin
The minimum graft copolymer concentration of bacterium growth.
By above-mentioned experiment, following experimental result is obtained:
After amphipathic graft copolymer G-EPL and Escherichia coli effect (experimental group), compared to amphipathic grafting is not added
The bacterium colony concentration of the pure Escherichia coli (control group) of copolymer G-EPL is reduced, and illustrates amphipathic graft copolymer G-
EPL can effectively inhibit the growth of Escherichia coli.Within the scope of effective Mlc, with amphipathic graft copolymer G-
The concentration of EPL increases, and experimental group culture dish colony number significantly reduces.
After amphipathic graft copolymer G-melittin and staphylococcus aureus effect (experimental group), compared to not being added
The bacterium colony concentration of the proof gold staphylococcus aureus (control group) of amphipathic graft copolymer G-melittin is reduced, and is said
Bright amphipathic graft copolymer G-melittin can effectively inhibit the growth of staphylococcus aureus.In effective Mlc
In range, as the concentration of amphipathic graft copolymer G-melittin increases, experimental group culture dish colony number is significantly reduced.
It is amphipathic compared to not being added after amphipathic graft copolymer G- (LL-37) and Escherichia coli act on (experimental group)
The bacterium colony concentration of the pure Escherichia coli (control group) of graft copolymer G- (LL-37) is reduced, and illustrates amphipathic grafting
Copolymer G- (LL-37) can effectively inhibit the growth of Escherichia coli.Within the scope of effective Mlc, connect with amphipathic
The concentration of graft copolymer G- (LL-37) increases, and experimental group culture dish colony number significantly reduces.
By antibiotic property test in it can be seen from good amphipathic containing antibacterial peptide of biocompatibility prepared by the present invention connect
Graft copolymer (G-EPL, G-melittin and G- (LL-37)) has excellent restraining and sterilizing bacteria effect, and (leather is blue to Escherichia coli
Family name's negative bacterium) and staphylococcus aureus (gram-positive bacteria) all have good antibacterial effect, therefore, the present invention preparation
The good amphipathic graft copolymer containing antibacterial peptide of biocompatibility have excellent broad spectrum antibacterial performance.
The above-mentioned description to embodiment is that this hair can be understood and used for the ease of those skilled in the art
It is bright.Those skilled in the art obviously readily can make various modifications to these embodiments, and described herein one
As principle be applied in other embodiments, without having to go through creative labor.Therefore, the present invention is not limited to the above embodiments.
Those skilled in the art's principle according to the present invention, not departing from improvement that scope of the invention is made and modification all should be at this
Within the protection scope of invention.
Claims (9)
1. a kind of amphipathic graft copolymer containing antibacterial peptide, it is characterised in that: its structural formula is as follows:
Wherein, integer of the J in 1-50, integer of the K in 1-50, integer of the L in 1-50；J, K and L are respectively represented
The degree of polymerization of random copolymerization block in graft copolymer, Pep indicate antibacterial peptide.
2. the amphipathic graft copolymer according to claim 1 containing antibacterial peptide, it is characterised in that: the antibacterial peptide is selected from
One or more of melittin, epsilon-polylysine, LL-37, bacitracin and cecropin；And/or
The hydrophobic polymer is poly- (6-caprolactone) of the carboxyl functional group of second containing side.
3. a kind of preparation method of the amphipathic graft copolymer according to claim 1 containing antibacterial peptide, it is characterised in that:
It includes the following steps:
(1), using methoxy poly (ethylene glycol) as initiator, stannous iso caprylate is catalyst, by the ε-of 6-caprolactone and function dough
Ring opening copolymer synthesis contains side acetic acid benzyl ester base to caprolactone monomer 6- (acetic acid benzyl ester base -6-caprolactone) in organic solvent
6-caprolactone/6- (acetic acid benzyl ester base)-epsilon-caprolactone copolymer, and slough benzyl protection under palladium carbon catalytic hydrogenation conditions
Group obtains hydrophobic polymer, structural formula are as follows:
Wherein, J and K respectively represents the degree of polymerization of each random copolymerization block in hydrophobic polymer；
(2), by the hydrophobic polymer, N, N '-dicyclohexylcarbodiimide and n-hydroxysuccinimide are in organic solvent
Interior reaction obtains the hydrophobic polymer segment that carboxyl is activated, structural formula are as follows:
Wherein, J and K respectively represents the degree of polymerization of each random copolymerization block in hydrophobic polymer；
(3), the hydrophobic polymer segment that the carboxyl is activated is dissolved in organic solvent, and antibacterial peptide is added and carries out instead
It answers, after washed, precipitating, drying, obtains the amphipathic graft copolymer containing antibacterial peptide, structural formula are as follows:
Wherein, integer of the J in 1-50, integer of the K in 1-50, integer of the L in 1-50, J, K and L are respectively represented
The degree of polymerization of random copolymerization block in graft copolymer, Pep indicate antibacterial peptide.
4. preparation method according to claim 3, it is characterised in that: the antibacterial peptide be selected from melittin, epsilon-polylysine,
One or more of LL-37, bacitracin and cecropin；And/or
The hydrophobic polymer is poly- (6-caprolactone) of the carboxyl functional group of second containing side.
5. preparation method according to claim 3, it is characterised in that: described in step (1), step (2) and step (3)
Organic solvent is selected from tetrahydrofuran, N,N-dimethylformamide, methylene chloride, toluene, p-methyl benzenesulfonic acid, morpholine, acetone
With any one or a few in dimethyl sulfoxide.
6. preparation method according to claim 3, it is characterised in that: in step (2), hydrophobicity that the carboxyl is activated
The molar ratio of polymer segment and the antibacterial peptide is 1:(1-100).
7. preparation method according to claim 3, it is characterised in that: the temperature of the reaction of the step (2) is -10-60
DEG C, the time of reaction is 1-24h.
8. preparation method according to claim 3, it is characterised in that: the temperature of the reaction of the step (3) is -10-60
DEG C, the time of reaction is 1-24h.
9. a kind of amphipathic graft copolymer containing antibacterial peptide as described in claim 1 is facing as antibacterial agent or pharmaceutical carrier
Application in terms of anti-infective, the anticancer drug Targeting delivery of bed or nanosecond medical science.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910608080.1A CN110330656A (en) | 2019-07-08 | 2019-07-08 | Amphipathic graft copolymer and its preparation method and application containing antibacterial peptide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910608080.1A CN110330656A (en) | 2019-07-08 | 2019-07-08 | Amphipathic graft copolymer and its preparation method and application containing antibacterial peptide |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110330656A true CN110330656A (en) | 2019-10-15 |
Family
ID=68143921
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910608080.1A Pending CN110330656A (en) | 2019-07-08 | 2019-07-08 | Amphipathic graft copolymer and its preparation method and application containing antibacterial peptide |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110330656A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114159461A (en) * | 2021-11-23 | 2022-03-11 | 同济大学 | Penicillin-modified epsilon-polylysine antibacterial peptide and preparation method thereof |
| CN114805821A (en) * | 2022-04-28 | 2022-07-29 | 上海摩漾生物科技有限公司 | Degradable antibacterial peptide based on cellulose and preparation method thereof |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102423294A (en) * | 2011-12-23 | 2012-04-25 | 天津大学 | Thermo sensitive in situ gel loaded with drug by chemical bonds, and preparation method thereof |
| CN103221070A (en) * | 2010-08-30 | 2013-07-24 | 哈佛大学校长及研究员协会 | Shear controlled release for stenotic lesions and thrombolytic therapies |
| CN105147615A (en) * | 2015-08-17 | 2015-12-16 | 上海市肿瘤研究所 | Tumor cell and tumor vessel double-target nanoparticle, building method and application |
| CN106750262A (en) * | 2016-11-30 | 2017-05-31 | 同济大学 | The synthesis of amphipathic block antibacterial peptide and its preparation method and application of assembly |
| CN106943599A (en) * | 2017-02-09 | 2017-07-14 | 南京鼓楼医院 | A kind of preparation method for the nanometer medicine-carried system for carrying chemical anti-tumor drugs and anti Fas antibody altogether |
| CN107141492A (en) * | 2017-06-08 | 2017-09-08 | 华南理工大学 | One kind has target function echovirus structuring polymer vesica and its preparation and application |
| CN107997984A (en) * | 2017-11-27 | 2018-05-08 | 肽源（广州）生物科技有限公司 | A kind of antibacterial peptide-chitosan complexes, its preparation method and its application |
| CN108815552A (en) * | 2018-07-05 | 2018-11-16 | 四川大学 | A kind of drug controllably loads and the bio-medical coating material of long-acting slow-release and preparation method thereof |
-
2019
- 2019-07-08 CN CN201910608080.1A patent/CN110330656A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103221070A (en) * | 2010-08-30 | 2013-07-24 | 哈佛大学校长及研究员协会 | Shear controlled release for stenotic lesions and thrombolytic therapies |
| CN102423294A (en) * | 2011-12-23 | 2012-04-25 | 天津大学 | Thermo sensitive in situ gel loaded with drug by chemical bonds, and preparation method thereof |
| CN105147615A (en) * | 2015-08-17 | 2015-12-16 | 上海市肿瘤研究所 | Tumor cell and tumor vessel double-target nanoparticle, building method and application |
| CN106750262A (en) * | 2016-11-30 | 2017-05-31 | 同济大学 | The synthesis of amphipathic block antibacterial peptide and its preparation method and application of assembly |
| CN106943599A (en) * | 2017-02-09 | 2017-07-14 | 南京鼓楼医院 | A kind of preparation method for the nanometer medicine-carried system for carrying chemical anti-tumor drugs and anti Fas antibody altogether |
| CN107141492A (en) * | 2017-06-08 | 2017-09-08 | 华南理工大学 | One kind has target function echovirus structuring polymer vesica and its preparation and application |
| CN107997984A (en) * | 2017-11-27 | 2018-05-08 | 肽源（广州）生物科技有限公司 | A kind of antibacterial peptide-chitosan complexes, its preparation method and its application |
| CN108815552A (en) * | 2018-07-05 | 2018-11-16 | 四川大学 | A kind of drug controllably loads and the bio-medical coating material of long-acting slow-release and preparation method thereof |
Non-Patent Citations (3)
| Title |
|---|
| ZHENGZHEN DU，等: "Synthesis of functionalized Pluronic-b-poly(e-caprolactone) and the comparative study of their pendant groups on the cellular internalization behavior", 《J MATER SCI: MATER MED》 * |
| ZHOU, XY，等: "trategies from nature: polycaprolactone-based mimetic antimicrobial peptide block copolymers with low cytotoxicity and excellent antibacterial efficiency", 《POLYMER CHEMISTRY》 * |
| 孙琰，等: "含侧羧基官能团的聚(ε-己内酯)降解性能的研究", 《化学学报》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114159461A (en) * | 2021-11-23 | 2022-03-11 | 同济大学 | Penicillin-modified epsilon-polylysine antibacterial peptide and preparation method thereof |
| CN114805821A (en) * | 2022-04-28 | 2022-07-29 | 上海摩漾生物科技有限公司 | Degradable antibacterial peptide based on cellulose and preparation method thereof |
| CN114805821B (en) * | 2022-04-28 | 2023-02-03 | 上海摩漾生物科技有限公司 | Degradable antibacterial peptide based on cellulose and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US10494451B2 (en) | Chitosan-derivative compounds and methods of controlling microbial populations | |
| Hu et al. | Supramolecular hydrogels for antimicrobial therapy | |
| CN101909666B (en) | Antimicrobial coatings for medical devices and methods for making and using them | |
| ES2375524T3 (en) | ANTIMICROBIAL MEDICAL-TECHNICAL PRODUCT, PROCEDURE FOR MANUFACTURING AND USE. | |
| Liu et al. | Antibacterial PCL electrospun membranes containing synthetic polypeptides for biomedical purposes | |
| Qiu et al. | Peptidoglycan-inspired peptide-modified injectable hydrogels with enhanced elimination capability of bacterial biofilm for chronic wound healing | |
| CN107513145A (en) | The synthesis of biodegradable amphiphilic block class antibacterial peptide and the preparation method and application of class antibacterial peptide vesica | |
| CN110330656A (en) | Amphipathic graft copolymer and its preparation method and application containing antibacterial peptide | |
| CN110527080A (en) | Amphipathic multi-block class antibacterial peptide copolymer and its preparation method and application | |
| JP5247144B2 (en) | Polymer coupling agents and pharmaceutically active polymers made therefrom | |
| Liu et al. | A polypeptide coating for preventing biofilm on implants by inhibiting antibiotic resistance genes | |
| CN106279635B (en) | Class antibacterial peptide and vesica and its preparation method and application | |
| AU2018264151B2 (en) | Chitosan-Derivative Compounds and Methods of Controlling Microbial Populations | |
| CN110054713A (en) | Amphipathic graft copolymer and its preparation method and application containing antibacterial peptide | |
| CN111234163B (en) | Nanogel with antibacterial repair performance and preparation method and application thereof | |
| AU2013211460B2 (en) | Chitosan-Derivative Compounds and Methods of Controlling Microbial Populations | |
| US20170202217A1 (en) | Antibacterial polymer, production method therefor, and usage thereof | |
| CN113234215B (en) | Cationic cholic acid polyester antibacterial peptide and preparation method and application thereof | |
| CN113801568B (en) | Antibacterial coating with pH response function, functional material with antibacterial coating and preparation method thereof | |
| Krivkina et al. | Hemocompatibility and cytotoxicity of small-diameter bioabsorbable tissue-engineered vascular grafts depending on anti-thrombogenic and antimicrobial coating | |
| CN105778127A (en) | Polylactic acid-polypeptide polymeric micelle and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20191015 |