CN106466475B - Composite formed by combining antibacterial peptide and polymer, preparation method and application thereof - Google Patents

Composite formed by combining antibacterial peptide and polymer, preparation method and application thereof Download PDF

Info

Publication number
CN106466475B
CN106466475B CN201510518175.6A CN201510518175A CN106466475B CN 106466475 B CN106466475 B CN 106466475B CN 201510518175 A CN201510518175 A CN 201510518175A CN 106466475 B CN106466475 B CN 106466475B
Authority
CN
China
Prior art keywords
polymer
poly
polyethylene glycol
acid
complex
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.)
Active
Application number
CN201510518175.6A
Other languages
Chinese (zh)
Other versions
CN106466475A (en
Inventor
刘克良
王晨宏
郄建坤
冯思良
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Institute of Pharmacology and Toxicology of AMMS
Original Assignee
Institute of Pharmacology and Toxicology of AMMS
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Institute of Pharmacology and Toxicology of AMMS filed Critical Institute of Pharmacology and Toxicology of AMMS
Priority to CN201510518175.6A priority Critical patent/CN106466475B/en
Priority to PCT/CN2016/095471 priority patent/WO2017032236A1/en
Publication of CN106466475A publication Critical patent/CN106466475A/en
Application granted granted Critical
Publication of CN106466475B publication Critical patent/CN106466475B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/08—Peptides having 5 to 11 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/10—Peptides having 12 to 20 amino acids
    • 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
    • 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
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Epidemiology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Inorganic Chemistry (AREA)
  • Medicinal Preparation (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Peptides Or Proteins (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)

Abstract

The invention relates to a compound formed by combining antibacterial peptide and polymer, a preparation method of the compound and application of the compound in preparing a medicament for preventing or treating diseases which can be prevented or treated by the antibacterial peptide. The compound of the invention maintains the activity of the antibacterial peptide, simultaneously, the hemolytic toxicity is greatly reduced, and the invention has good application prospect.

Description

Composite formed by combining antibacterial peptide and polymer, preparation method and application thereof
Technical Field
The invention relates to a compound formed by combining antibacterial peptide and polymer, a preparation method of the compound and application of the compound in preparing a medicament for preventing or treating diseases which can be prevented or treated by the antibacterial peptide.
Background
The antibacterial peptide is considered to be developed into a new generation of antibiotic drugs because of its advantages of broad-spectrum antibacterial activity and low susceptibility to drug resistance. This is due to their different bactericidal mechanisms than current antibiotic drugs: most of antibacterial peptide sequences contain more basic amino acids, so that the antibacterial peptide sequences are rich in positive charges and easy to be electrostatically attracted with negatively charged phospholipids on cell membranes, and then cationic polypeptides are inserted into bilayers of the cell membranes to form holes, so that intracellular fluid is leaked, and bacteria are killed.
Despite the high antimicrobial activity, no antimicrobial peptide drug has been approved for sale from the first antimicrobial peptide discovery to date. The biggest obstacle is that the cationic antibacterial peptide has strong hemolytic side effects and other side effects besides the difficulties of common polypeptides in drug development (such as the defects that polypeptide compounds are easy to combine with serum and plasma in vivo and are degraded by proteolytic enzyme, the half-life period in vivo is short, the stability is poor, the antigenicity is strong, and the like). Therefore, the antimicrobial peptides require higher doses for use, which in turn leads to hemolytic toxicity. Therefore, how to obtain stable, safe and effective antibacterial peptide drugs or preparations has become a research hotspot with great challenge and practical value.
Disclosure of Invention
The present inventors have diligently made efforts and extensive experiments, and surprisingly found that a polyion complex (Polyionic complex) prepared by using an electrostatic interaction between an anionic polymer and a cationic antimicrobial peptide can greatly reduce its hemolytic toxicity while maintaining the activity of the antimicrobial peptide, thereby completing the present invention.
The present invention relates in a first aspect to a complex comprising an antimicrobial peptide and a polymer, wherein the antimicrobial peptide is positively charged and the polymer is negatively charged.
In an embodiment of the invention, the antimicrobial peptide and the polymer are bound by electrostatic interaction.
In an embodiment of the invention, the antimicrobial peptide is a cationic antimicrobial peptide.
In an embodiment of the present invention, the antimicrobial peptide is one or more selected from the group consisting of a polypeptide, a derivative thereof, a pharmaceutically acceptable salt thereof, and a D-isomer thereof.
In an embodiment of the present invention, the cationic antibacterial peptide is a cationic antibacterial peptide with 5-20 positive charges, for example, any one selected from pexiganan, omiganan, hLF1-11, P113, XMP629, or a mixture thereof.
Wherein, the polypeptide sequences of the antibacterial peptides are respectively as follows:
pexiganan: GIGKF LKKAK KFGKA FVKIL KK-NH2
Omega plus south: ILRWP WWPWR RK-NH2
LF1-11：GRRRR RSVQW CA
P113：AKRHH GYKRK FH-NH2
XMP629：KLFR-(3-(1-naphthyl)-A-QAK-(3-(1-naphthyl)-A-NH2
In one embodiment of the invention, the cationic antimicrobial peptide is pexiganan.
In another embodiment of the invention, the cationic antimicrobial peptide is omiganan.
In embodiments of the present invention, wherein the polymer has good biocompatibility and is biodegradable, it can be used in vivo as a pharmaceutical carrier, an adjuvant, an excipient, etc.;
in an embodiment of the present invention, the polymer contains a hydrophilic segment or both a hydrophilic segment and a hydrophobic segment, preferably, the hydrophilic segment is located at the N-terminus of the hydrophobic segment,
wherein the hydrophilic chain segment contains a polymer of acidic amino acid or its derivative, or the hydrophilic chain segment contains a copolymer of acidic amino acid and hydrophilic amino acid such as serine and threonine or its derivative, such as a copolymer of acidic amino acid and serine or its derivative, or a copolymer of acidic amino acid and threonine or its derivative, optionally, the N-terminal of the acidic amino acid is also connected with a molecule capable of initiating amino acid polymerization, such as polyethylene glycol, serine, threonine, C1-10Alkyl (e.g. C)1-6Alkyl) or other small molecular weight compounds (molecular weight less than 1000) that can initiate the polymerization of amino acids,
a polymer comprising a hydrophobic amino acid or a derivative thereof in the hydrophobic segment.
In an embodiment of the invention, the acidic amino acid comprises glutamic acid, aspartic acid.
In one embodiment of the present invention, the hydrophilic segment is polyglutamic acid.
In one embodiment of the present invention, the hydrophilic segment is a (glutamic acid-aspartic acid) copolymer.
In one embodiment of the present invention, the hydrophilic segment is polyethylene glycol-polyglutamic acid.
In one embodiment of the present invention, the hydrophilic segment is a polyethylene glycol- (glutamic acid-aspartic acid) copolymer.
In one embodiment of the present invention, the hydrophilic segment is a (glutamic acid-aspartic acid-serine) copolymer.
In one embodiment of the present invention, the hydrophilic segment is a polyethylene glycol- (glutamic acid-aspartic acid-serine) copolymer.
In one embodiment of the invention, the hydrophobic segment is poly-isoleucine.
In one embodiment of the invention, the hydrophobic segment is poly-naphthylalanine.
In an embodiment of the invention, the polyethylene glycol is PEG or mPEG.
In an embodiment of the present invention, the molecular weight of the polyethylene glycol is 600-.
In an embodiment of the invention, the polymer is a compound of formula i:
Figure BDA0000785761470000031
wherein R is1Absent, or a molecule which can initiate the polymerization of an amino acid, for example selected from polyethylene glycol (polyethylene glycol monomethyl ether or polyethylene glycol), serine, threonine, C1-10Alkyl (e.g. C)1-6Alkyl), or other small molecular weight compounds (molecular weight less than 1000) that can initiate amino acid polymerization;
R2is selected from- (CH)2)xCOOH and pharmaceutically acceptable salts thereof, wherein x ═ 0 to 5 (e.g., 0,1, 2, 3, 4, 5);
R3is selected from hydrophobic amino acid residues, e.g. selected from Ile, Leu, Phe, Pro, Val, Trp;
m is 5-100, and R2May be the same or different;
n is 0-100, and when n ≧ 2, R3May be the same or different.
In an embodiment of the present invention, the molecular weight of the polyethylene glycol is 600-.
In an embodiment of the present invention, the hydrophobic amino acid derivative is, for example, naphthylalanine, benzyl glutamate, benzyl aspartate, or the like.
In an embodiment of the invention, said value of m is from 15 to 51.
In an embodiment of the invention, the value of n is 0 or n is 5 to 19.
The composite according to any one of the first aspect of the invention, wherein the polymer is selected from one or several of hydrophilic anionic polymers, amphiphilic diblock anionic polymers, amphiphilic anionic polymers;
preferably, the polymer is selected from polyglutamic acid, polyaspartic acid, (glutamic acid-aspartic acid) copolymer, polyethylene glycol-polyglutamic acid, polyethylene glycol-polyaspartic acid, polyethylene glycol- (glutamic acid-aspartic acid) copolymer, polyglutamic acid-polyphenylalanine, polyglutamic acid-polylvaline, polyglutamic acid-polyleucine, polyglutamic acid-polyileucin, polyglutamic acid-polynaphthylalanine, polyaspartic acid-polyphenylalanine, polyaspartic acid-polyvaline, polyaspartic acid-polyleucine, polyaspartic acid-polynaphthylalanine, (glutamic acid-aspartic acid) copolymer-polyphenylalanine, (glutamic acid-aspartic acid) copolymer-polyvaline, poly (glutamic acid-aspartic acid) copolymer-poly (glutamic acid-aspartic acid), (glutamic acid-aspartic acid) copolymer-poly leucine, (glutamic acid-aspartic acid) copolymer-poly isoleucine, (glutamic acid-aspartic acid-serine) copolymer-poly isoleucine, (glutamic acid-aspartic acid) copolymer-poly naphthylalanine, polyethylene glycol-poly glutamic acid-poly leucine, polyethylene glycol-poly glutamic acid-poly valine, polyethylene glycol-poly glutamic acid-poly isoleucine, polyethylene glycol-poly glutamic acid-poly phenylalanine, polyethylene glycol-poly glutamic acid-poly naphthylalanine, polyethylene glycol-poly aspartic acid-poly valine, polyethylene glycol-poly aspartic acid-poly leucine, polyethylene glycol-poly aspartic acid-poly isoleucine, poly (glutamic acid-poly (aspartic acid-poly (isoleucine)), (glutamic acid-poly (aspartic acid-poly (naphthylalanine)), (glutamic acid-poly, Polyethylene glycol-polyaspartic acid-poly (phenylalanine), polyethylene glycol-polyaspartic acid-poly (naphthylalanine), polyethylene glycol- (glutamic acid-aspartic acid) copolymer-poly (phenylalanine), polyethylene glycol- (glutamic acid-aspartic acid) copolymer-poly (valine), one or more of polyethylene glycol- (glutamic acid-aspartic acid) copolymer-poly leucine, polyethylene glycol- (glutamic acid-aspartic acid) copolymer-poly isoleucine, polyethylene glycol- (glutamic acid-aspartic acid-serine) copolymer-poly isoleucine, polyethylene glycol- (glutamic acid-aspartic acid) copolymer-poly naphthylalanine, wherein amino acids in the hydrophilic chain segment can be arranged in any order. Preferably, the number of amino acid repeat units in the hydrophilic segment is from 5 to 100, preferably from 15 to 60. Preferably, the number of hydrophobic amino acid repeat units is from 0 to 100, preferably from 5 to 19.
In an embodiment of the present invention, the molecular weight of the polyethylene glycol ranges from 1000-.
In one embodiment of the invention, the polymer is polyethylene glycol-polyglutamic acid; in another embodiment of the present invention, the polymer is polyethylene glycol-polyglutamic acid-poly-isoleucine.
The complex according to any one of the first aspect of the invention, wherein the molar ratio of polymer to antimicrobial peptide is from 0.02:1 to 50:1, such as from 0.04:1 to 15:1, such as from 0.2:1 to 5: 1.
In an embodiment of the invention, wherein the molar ratio of polymer to antimicrobial peptide is 0.04:1, 0.2:1, 0.3:1, 1:1, 3:1, 5: 1.
In an embodiment of the invention, the complex forms a nanomicelle structure in solution; for example, when the polymer is a triblock polymer, such as a polyethylene glycol-acidic amino acid polymer-hydrophobic polymer segment, the hydrophobic amino acid polymer is located inside the micelle, and the acidic amino acid polymer and the antibacterial peptide are outside the hydrophobic amino acid polymer, and the outermost is a molecule capable of initiating amino acid polymerization, such as polyethylene glycol.
In an embodiment of the invention, the positively charged antimicrobial peptide electrostatically interacts with negatively charged groups in the polymer, e.g. amino groups in the antimicrobial peptide electrostatically interact with carboxyl groups in hydrophilic segments (e.g. acidic amino acids) in the polymer.
A second aspect of the invention relates to a pharmaceutical composition comprising a complex according to any one of the first aspects of the invention, together with a pharmaceutically acceptable carrier or excipient.
A third aspect of the present invention relates to the use of a complex according to any one of the first aspect of the present invention in the manufacture of a medicament for the prophylaxis or treatment of a disease for which an antimicrobial peptide is indicated.
The use according to any of the third aspect of the present invention, wherein the disease which the antibacterial peptide can prevent or treat is a disease caused by bacteria (e.g. gram-positive or gram-negative bacteria), fungi or viruses; preferably, the bacteria, fungi or viruses are bacteria, fungi or viruses that the antimicrobial peptide is capable of inhibiting or killing.
For example, pexiganan can be used for preventing or treating infection caused by gram-positive bacteria or gram-negative bacteria, and the compound of the invention can be used for preparing a medicament for preventing or treating infection caused by gram-positive bacteria or gram-negative bacteria.
A fourth aspect of the invention relates to the use of a complex according to any one of the first aspect of the invention for inhibiting or killing bacteria (e.g. gram positive or gram negative), fungi or viruses in vitro/in vivo; preferably, the bacteria, fungi or viruses are bacteria, fungi or viruses that the antimicrobial peptide is capable of inhibiting or killing.
A fifth aspect of the present invention relates to a method for preparing a complex according to any one of the first aspect of the present invention, comprising the steps of:
respectively dissolving the antibacterial peptide and the polymer in water according to a certain proportion, stirring to fully mix the antibacterial peptide and the polymer, then separating (removing substances which are not combined) to obtain the compound, and optionally, further comprising a step of freeze-drying after separation;
preferably, the molar ratio of the polymer to the antibacterial peptide is 0.02: 1-50: 1, such as 0.04: 1-15: 1, such as 0.2: 1-5: 1.
In an embodiment of the invention, the antimicrobial peptide is a cationic antimicrobial peptide.
In an embodiment of the present invention, the antimicrobial peptide is one or more selected from the group consisting of a polypeptide, a derivative thereof, a pharmaceutically acceptable salt thereof, and a D-isomer thereof.
For example, the cationic antibacterial peptide is a cationic antibacterial peptide with 5-20 positive charges, such as any one selected from pexiganan, omiganan, hLF1-11, P113, XMP629, or a mixture of the above.
In an embodiment of the invention, the cationic antimicrobial peptide is pexiganan.
In another embodiment of the invention, the cationic antimicrobial peptide is omiganan.
In an embodiment of the invention, the polymer is an anionic polymer.
In the embodiment of the invention, wherein the polymer has good biocompatibility, can be used as a drug carrier, an auxiliary agent, an excipient and the like in vivo;
in an embodiment of the present invention, the polymer contains a hydrophilic segment or both a hydrophilic segment and a hydrophobic segment, preferably, the hydrophilic segment is located at the N-terminus of the hydrophobic segment,
wherein the hydrophilic chain segment contains a polymer of acidic amino acid or its derivative, or the hydrophilic chain segment contains a copolymer of acidic amino acid and hydrophilic amino acid such as serine and threonine or its derivative, such as a copolymer of acidic amino acid and serine or its derivative, or a copolymer of acidic amino acid and threonine or its derivative, optionally, the N-terminal of the acidic amino acid is also connected with a molecule capable of initiating amino acid polymerization, such as polyethylene glycol, serine, threonine, C1-10Alkyl (e.g. C)1-6Alkyl) or other small molecular weight compounds (molecular weight less than 1000) that can initiate the polymerization of amino acids,
a polymer comprising a hydrophobic amino acid or a derivative thereof in the hydrophobic segment.
In an embodiment of the invention, the acidic amino acid comprises glutamic acid, aspartic acid.
In one embodiment of the present invention, the hydrophilic segment is polyglutamic acid.
In one embodiment of the present invention, the hydrophilic segment is a (glutamic acid-aspartic acid) copolymer.
In one embodiment of the present invention, the hydrophilic segment is polyethylene glycol-polyglutamic acid.
In one embodiment of the present invention, the hydrophilic segment is a polyethylene glycol- (glutamic acid-aspartic acid) copolymer.
In one embodiment of the present invention, the hydrophilic segment is a (glutamic acid-aspartic acid-serine) copolymer.
In one embodiment of the present invention, the hydrophilic segment is a polyethylene glycol- (glutamic acid-aspartic acid-serine) copolymer.
In one embodiment of the invention, the hydrophobic segment is poly-isoleucine.
In one embodiment of the invention, the hydrophobic segment is poly-naphthylalanine.
In an embodiment of the present invention, the molecular weight of the polyethylene glycol is 600-.
In an embodiment of the invention, the polymer is a compound of formula i:
Figure BDA0000785761470000081
wherein R is1Absent, or a molecule which can initiate the polymerization of an amino acid, for example selected from polyethylene glycol (polyethylene glycol monomethyl ether or polyethylene glycol), serine, threonine, C1-10Alkyl (e.g. C)1-6Alkyl), or other small molecular weight compounds (molecular weight less than 1000) that can initiate amino acid polymerization;
R2is selected from- (CH)2)xCOOH and pharmaceutically acceptable salts thereof, wherein x ═ 0 to 5 (e.g., 0,1, 2, 3, 4, 5);
R3is selected from hydrophobic amino acid residues, e.g. selected from Ile, Leu, Phe, Pro, Val, Trp;
m is 5-100, and R2May be the same or different;
n is 0-100, and when n ≧ 2, R3May be the same or different.
In an embodiment of the present invention, the molecular weight of the polyethylene glycol is 600-.
In an embodiment of the present invention, the hydrophobic amino acid derivative is, for example, naphthylalanine, benzyl glutamate, benzyl aspartate, or the like.
In an embodiment of the invention, said value of m is from 15 to 51.
In an embodiment of the invention, the value of n is 0 or n is 5 to 19.
In an embodiment of the present invention, wherein said polymer is selected from one or several of hydrophilic anionic polymers, amphiphilic diblock anionic polymers, amphiphilic anionic polymers;
preferably, the polymer is selected from polyglutamic acid, polyaspartic acid, (glutamic acid-aspartic acid) copolymer, polyethylene glycol-polyglutamic acid, polyethylene glycol-polyaspartic acid, polyethylene glycol- (glutamic acid-aspartic acid) copolymer, polyglutamic acid-polyphenylalanine, polyglutamic acid-polylvaline, polyglutamic acid-polyleucine, polyglutamic acid-polyileucin, polyglutamic acid-polynaphthylalanine, polyaspartic acid-polyphenylalanine, polyaspartic acid-polyvaline, polyaspartic acid-polyleucine, polyaspartic acid-polynaphthylalanine, (glutamic acid-aspartic acid) copolymer-polyphenylalanine, (glutamic acid-aspartic acid) copolymer-polyvaline, poly (glutamic acid-aspartic acid) copolymer-poly (glutamic acid-aspartic acid), (glutamic acid-aspartic acid) copolymer-poly leucine, (glutamic acid-aspartic acid) copolymer-poly isoleucine, (glutamic acid-aspartic acid) copolymer-poly naphthylalanine, (glutamic acid-aspartic acid-serine) copolymer-poly isoleucine, polyethylene glycol-poly glutamic acid-poly leucine, polyethylene glycol-poly glutamic acid-poly valine, polyethylene glycol-poly glutamic acid-poly isoleucine, polyethylene glycol-poly glutamic acid-poly phenylalanine, polyethylene glycol-poly glutamic acid-poly naphthylalanine, polyethylene glycol-poly aspartic acid-poly valine, polyethylene glycol-poly aspartic acid-poly leucine, polyethylene glycol-poly aspartic acid-poly isoleucine, poly naphthylalanine, poly naphthylamine, Polyethylene glycol-polyaspartic acid-poly (phenylalanine), polyethylene glycol-polyaspartic acid-poly (naphthylalanine), polyethylene glycol- (glutamic acid-aspartic acid) copolymer-poly (phenylalanine), polyethylene glycol- (glutamic acid-aspartic acid) copolymer-poly (valine), one or more of polyethylene glycol- (glutamic acid-aspartic acid) copolymer-poly leucine, polyethylene glycol- (glutamic acid-aspartic acid) copolymer-poly isoleucine, polyethylene glycol- (glutamic acid-aspartic acid-serine) copolymer-poly isoleucine and polyethylene glycol- (glutamic acid-aspartic acid) copolymer-poly naphthylalanine, wherein amino acids in the hydrophilic chain segment can be arranged in any order. Preferably, the number of amino acid repeat units in the hydrophilic segment is from 5 to 100, preferably from 15 to 60. Preferably, the number of hydrophobic amino acid repeat units is from 0 to 100, preferably from 5 to 19.
In an embodiment of the present invention, the molecular weight of the polyethylene glycol ranges from 1000-.
In one embodiment of the invention, the polymer is polyethylene glycol-polyglutamic acid; in another embodiment of the present invention, the polymer is polyethylene glycol-polyglutamic acid-poly-isoleucine.
The invention also relates to the use of a polymer for the preparation of a pharmaceutical formulation of an antimicrobial peptide, wherein the polymer is negatively charged.
In the invention, the antibacterial peptides (antibacterial peptides) refer to a class of small molecular polypeptides with antibacterial activity in various biological natural immune systems, and also include simulated peptides artificially synthesized by taking natural antibacterial peptides as templates.
In the present invention, the cationic antibacterial peptides (cationic antibacterial peptides) refer to amphipathic molecules generally composed of 12-50 amino acid residues, which are generated by plants and animals, exist in organisms, have functions of resisting external microbial invasion and eliminating in-vivo mutant cells, and also include mimic peptides artificially synthesized by using natural cationic antibacterial peptides as templates.
In the present invention, the polymer is an anionic polymer, i.e., a polymer containing a negative charge.
In the present invention, the electrostatic interaction refers to attraction between opposite charges and repulsion between the same charges.
In the present invention, the binding of the antimicrobial peptide to the polymer by electrostatic action means electrostatic attraction between the carboxyl ion of the anionic polymer and the amino ion of the cationic polypeptide.
In the present invention, the polymer of amino acids is also referred to as polyamino acid.
In the present invention, the hydrophobic amino acid residue refers to a portion of the hydrophobic amino acid remaining after removal of a group forming a peptide bond, and for example, the leucine residue is an isobutyl group, and the phenylalanine residue is a benzyl group.
In the present invention, amino acids are all in the L form unless otherwise specified.
Drawings
Gel exclusion chromatogram of the polymer of FIG. 1
FIG. 2 nuclear magnetic spectrum of PEG-PGA (A) and PEG-PGA (B)
FIG. 3 schematic of the preparation of the complexes
FIG. 4 particle size distribution diagram (A) and Transmission Electron micrograph (B) of Complex 1
FIG. 5 gel electrophoresis of complexes
FIG. 6 nuclear magnetic spectra of PEG-PGM (A), PEG-PGM (B), and PEG-PGM (C)
FIG. 7 agarose gel electrophoresis of the complexes
FIGS. 8 to 12 are the results of the hemolytic activity test of pexiganan and each compound thereof
Detailed Description
Embodiments of the present invention will be described in detail below with reference to examples, but those skilled in the art will appreciate that the following examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products commercially available.
EXAMPLE 1 anionic Polymer polyethylene glycol5000-polyglutamic acid49(mPEG5000-b-PGlu49) Preparation of
Mixing mPEG5000-NH2(from Kay Bio Inc.) in N, N-Dimethylformamide (DMF) 50 times the molar amount of the mixtureGlutamic acid-5-benzyl ester-N-carboxyanhydride (Glu (OBzl) -NCA, reference N.Nishiyama, et al, Langmuir 15(1999) 377-383), heated to 50 ℃, stirred for 24 hours, then the concentrated solution was poured into ether to precipitate white precipitate, filtered and dried in vacuum to obtain white powder mPEG5000-b-pglu (obzl), molecular weight Mn 12214, PDI 1.25 of the polymer as determined by gel exclusion chromatography (GPC), as shown in fig. 1, mPEG5000-b-PGlu(OBzl)49Curve of (2) and mPEG5000There was a clear difference indicating that a block polymer was obtained. Then, the resulting mixture was added to a 0.5N aqueous solution of sodium hydroxide to remove the protecting group, and the mixture was lyophilized to obtain a white powder. By passing1H NMR, comparison of polyethylene glycol CH2CH2The number of H in O and benzyl ester C in polyglutamic acid6H5The value of H is calculated to obtain that m is 49, and the product is mPEG5000-b-PGlu49。
Figure BDA0000785761470000111
Preparation route of Scheme1 polyethylene glycol monomethyl ether-b-polyglutamic acid
EXAMPLE 2 preparation of antimicrobial peptide and Polymer Complex 1 (Complex 1 for short)
Mixing mPEG5000-b-PGlu49And aqueous solution of pexiganan (synthesized by a microwave polypeptide synthesizer) at a molar ratio of 1:1, stirring at room temperature for 12 hours, dialyzing in a dialysis bag with molecular weight cutoff of 3500 for 24 hours, and lyophilizing to obtain compound 1 (shown in fig. 3). The particle size of composite 1 was tested using a laser particle sizer (DLS) to be 195.7 + -5.6 nm in solution with a particle size distribution of 0.12 + -0.03 (Table 1). Meanwhile, a Transmission Electron Microscope (TEM) as shown in FIG. 4 shows that the core-shell structure is obtained. Indicating that complex 1 is a stable nanomicelle.
Table 1 particle size of the composite and its dispersion index (n ═ 3).
Figure BDA0000785761470000121
EXAMPLE 3 preparation of Compounds of other ratios
Separately adding pexiganan and mPEG5000-b-PGlu49Compound 2, compound 3 and compound 4 were prepared according to the method of example 2 by mixing them at a molar ratio of 1:5, 1:0.2 and 1: 25.
Example 4 anionic Polymer mPEG5000Preparation of (b) -PGlu
Mixing mPEG5000-NH2Dissolving in DMF, adding Glu (OBzl) -NCA at different ratio, heating to 50 deg.C, stirring for 24 hr, pouring the concentrated solution into diethyl ether to precipitate white precipitate, filtering, and vacuum drying to obtain white powder. Then, the resulting mixture was added to a 0.5N aqueous solution of sodium hydroxide to remove the protecting group, and lyophilized. By passing1H NMR calculation determined the number of repeating units of glutamic acid to be m 15 and m 31, respectively, i.e., mPEG5000-b-PGlu15And mPEG5000-b-PGlu31。
EXAMPLE 5 preparation of antimicrobial peptides and Polymer complexes 5, 6 (Complex 5 and Complex 6 for short)
Mixing mPEG5000-b-PGlu15And mPEG5000-b-PGlu31Compound 5 and compound 6 were prepared according to the method of example 2 by mixing with pexiganan at a molar ratio of 1:1, respectively. The particle sizes and their particle size distributions are listed in table 1.
EXAMPLE 6 gel electrophoresis testing of complexes
In order to facilitate the observation of the formation of the complex, the invention uses the fluorescence labeled pexiganan instead of the pexiganan, which has the same number of positive charges and has no influence on the preparation of the complex. Mixing the compound 1-6, fluorescent labeled pexiganan, and fluorescent labeled pexiganan and polyethylene glycol5000The mixture of (1%) was added to a well of 1% agarose gel at a certain concentration, and the direction of movement of the fluorescent substance was observed after applying electricity of 20mV to both sides of the gel plate for 35 minutes. FIG. 5 shows fluorescently labeled pexiganan, fluorescently labeled pexiganan and polyethylene glycol5000The mixture of (a) moves toward the negative electrode and the other compound moves toward the positive electrode. The fluorescence intensity increased with increasing number of anions in the complex, indicating that pexiganan and peg-b-polyGlutamic acid forms a complex, and the stability improves with increasing number of anions.
Example 7 anionic Polymer polyethylene glycol5000-polyglutamic acid49-Poly-isoleucine (mPEG)5000-b-PGlu49Preparation of (b-PIle)
Mixing mPEG5000-b-PGlu49Dissolved in DMF, and isoleucine-N-carboxyanhydride (Ile-NCA, reference N. Nishiyama, et al, Langmuir 15(1999) 377-383) in different proportions was added thereto, heated to 50 ℃ and stirred for 24 hours, and the concentrated solution was poured into ether to precipitate a white precipitate, filtered, and vacuum-dried to obtain a white powder. Then, the resulting mixture was added to a 0.5N aqueous solution of sodium hydroxide to remove the protecting group, and the mixture was lyophilized to obtain a white powder.1H NMR detection, comparison of polyethylene glycol CH2CH2The number of hydrogens in O and CH in poly-isoleucine3CH2CH(CH3) And the numerical value of the hydrogen in the formula is calculated to obtain n-5 and n-13. I.e. mPEG5000-b-PGlu49-b-PIle5And mPEG5000-b-PGlu49-b-PIle13FIG. 6 shows a triblock polymer mPEG5000-b-PGlu49-b-PIle13Nuclear magnetic spectrum of (1).
Figure BDA0000785761470000131
Scheme 2 triblock Polymer mPEG5000-b-PGlu49-b-PIle13Synthetic route of (1)
EXAMPLE 8 preparation of antimicrobial peptides and Polymer complexes 7, 8 (Complex 7 and Complex 8 for short)
Mixing mPEG5000-b-PGlu49-b-PIle5And mPEG5000-b-PGlu49-b-PIle13Compound 7 and compound 8 were prepared according to the method of example 2, mixed with pexiganan at a molar ratio of 1:1, respectively.
Example 9 preparation of antimicrobial peptides and Polymer complexes 9, 10 (Complex 9 and Complex 10 for short)
Mixing mPEG5000-b-PGlu49-b-PIle13Compound 9 and compound 10 were prepared according to the method of example 2, mixed with pexiganan in a molar ratio of 1:3, 3:1, respectively.
Example 10 anionic Polymer mPEG2000-b-PGlu51-b-PIle19Preparation of
Mixing mPEG2000-NH2Dissolving in DMF, adding Glu (OBzl) -NCA, heating to 50 deg.C, stirring for 24 hr, concentrating, adding into diethyl ether to precipitate white precipitate, filtering, vacuum drying to obtain white powder,1h NMR measurement of glutamic acid repeating unit number m 51, i.e., mPEG2000-b-PGlu(OBzl)51. Dissolving in DMF, adding Ile-NCA, heating to 50 deg.C, stirring for 24 hr, concentrating, adding diethyl ether to precipitate white precipitate, filtering, vacuum drying to obtain white powder, and making into powder1H NMR calculated isoleucine repeat unit number n 19, i.e., mPEG2000-b-PGlu(OBzl)51-b-PIle19. Then, the resulting mixture was added to a 0.5N aqueous solution of sodium hydroxide to remove the protecting group, and the resulting mixture was lyophilized to obtain mPEG as a white powder2000-b-PGlu51-b-PIle19。
EXAMPLE 11 preparation of antimicrobial peptide and Polymer Complex 11 (Complex 11 for short)
Mixing mPEG2000-b-PGlu51-b-PIle19And pexiganan at a molar ratio of 1:1, and prepared according to the method of example 2 to give complex 11.
EXAMPLE 12 preparation of antimicrobial peptide and Polymer Complex 12 (Complex 12 for short)
Mixing mPEG2000-b-PGlu51-b-PIle19And pexiganan at a molar ratio of 1:5, and prepared according to the method of example 2 to give complex 12.
EXAMPLE 13 gel electrophoresis testing of complexes
In order to facilitate the observation of the formation of the complex, the invention uses the fluorescence labeled pexiganan instead of the pexiganan, which has the same number of positive charges and has no influence on the preparation of the complex. Combining the complexes 7-12, fluorescently labeled pexiganan, and fluorescently labeledPexiganan and polyethylene glycol5000The mixture of (1%) was added to a well of 1% agarose gel at a certain concentration, and the direction of movement of the fluorescent substance was observed after applying electricity of 20mV to both sides of the gel plate for 35 minutes. FIG. 7 shows fluorescently labeled pexiganan, fluorescently labeled pexiganan and polyethylene glycol5000The mixture of (a) moves toward the negative electrode and the other compound moves toward the positive electrode. The fluorescence intensity increased with increasing number of anions, indicating that pexiganan forms a complex with polyethylene glycol-b-polyglutamic acid-b-poly-isoleucine and the stability increased with increasing number of anions.
Example 14 determination of bacteriostatic Activity of Pexiganan and its complexes with polymers
Respectively inoculating experimental strains (gram positive bacteria, Staphylococcus aureus and Bacillus subtilis, gram negative bacteria, Escherichia coli) into the nutrient broth, culturing at 37 deg.C for 18-24 hr, and testing OD490The value is about 0.5, and the ratio of the raw materials is 1:10 before use5Diluting, taking 12 bacterial culture tubes and numbering. Preparing stock solution with the concentration of 1mg/mL (calculated by the amount of the pexiganan) of the pexiganan or the compound thereof, diluting by a dilution method, repeating the dilution for 3 times at each concentration, adding 100 mu L of bacterial liquid, gently shaking uniformly, culturing at 37 ℃ for 18-24 hours, and observing. After the culture, the culture tube is clear, and is still clear after shaking, and the tube is considered to grow aseptically; and (3) showing a turbid state to indicate the bacteria growth, and finding out a culture tube with the lowest concentration of the pexiganan from the tubes with the bacteria growth, wherein the concentration of the culture tube is the Minimum Inhibitory Concentration (MIC) for inhibiting the bacteria growth. The antibacterial results are shown in table 2, although the anionic polymer has no antibacterial activity, the complex shows obvious antibacterial activity on three common strains, and is equivalent to the antibacterial activity of the positive control pexiganan.
TABLE 2 Minimum Inhibitory Concentrations (MIC) of pexiganan and its complexes with polymers
Figure BDA0000785761470000161
Example 15 determination of hemolytic Activity of Pexiganan and complexes thereof with polymers
Preparing human blood red blood cells into a red blood cell suspension (prepared by diluting by 10 times), preparing pexiganan or a compound into stock solution with the concentration of 1mg/mL (calculated by the amount of the pexiganan), diluting by a dilution method, repeating the dilution method for three times for each sample, adding the red blood cell suspension, oscillating in an oscillator, centrifuging, taking supernatant, measuring the OD value at the wavelength of 414nm in a microplate reader, and performing 100% hemolysis on the red blood cells in Triton X-100 (positive control) by taking the red blood cells as 0 in PBS (negative control). The percent hemolysis was calculated by the following formula:
Figure BDA0000785761470000162
wherein A is the OD at 414 nm.
Fig. 8 to 12 are comparisons of hemolytic activities of pexiganan and complexes, and the results show that each complex significantly reduces hemolytic activity to human red blood cells.
Table 2 shows the therapeutic index (HC) of pexiganan and its complex10MIC) comparison [ Chen Y, Mant CT, Farmer SW, Hancock REW, Vasil ML, Hodges rs.j Biol Chem 2005; 280: 12316-29. ] Zhu WL, Nan YH, Hahm KS, Shin SY. J Biochem Mol Biol 2007; 1090-4 ], the therapeutic index is an important index for evaluating the safety of the medicine, and the larger the numerical value is, the safer the medicine is. As can be seen in Table 3, the therapeutic index of the complexes is improved, and the more the number of negative charges, the higher the therapeutic index.
Table 3 therapeutic index (HC) of pexiganan and its complexes10MIC) comparison
Figure BDA0000785761470000171
EXAMPLE 16 preparation of antimicrobial peptide and Polymer Complex 13 (Complex 13 for short)
Mixing mPEG5000-b-PGlu15And an aqueous solution of Omeganan (self-synthesized by microwave polypeptide synthesizer) at a molar ratio of 1:1, according to example 2The method prepares compound 13.
EXAMPLE 17 preparation of antimicrobial peptide and Polymer Complex 14 (Complex 14 for short)
Mixing mPEG5000-b-PGlu31And an aqueous solution of Omeganan (self-synthesized by a microwave polypeptide synthesizer) were mixed at a molar ratio of 1:1, and a complex 14 was prepared according to the method of example 2.
EXAMPLE 18 preparation of antimicrobial peptide and Polymer Complex 15 (Complex 15 for short)
Mixing mPEG5000-b-PGlu49And an aqueous solution of Omeganan (self-synthesized by a microwave polypeptide synthesizer) were mixed at a molar ratio of 1:1, and a complex 15 was prepared according to the method of example 2.
Example 19 measurement of the bacteriostatic Activity of Omeganan and complexes formed with polymers
The MIC of each complex was determined according to the method of example 14, and as a result, as shown in table 4, the complex showed significant bacteriostatic activity against three common strains, and was comparable to the positive control pexiomic bacteriostatic activity.
TABLE 4 Minimum Inhibitory Concentrations (MIC) of pexiganan and its complexes with polymers
Figure BDA0000785761470000181
Example 20 mouse acute toxicity test of Complex comprising antimicrobial peptide and Polymer
Taking 18-22 g of healthy balb/c male mice as experimental objects. Mice were randomly grouped by weight according to the principle of random grouping. 8-10 mice were set per group. Different doses of the drug (pexiganan group, 25,30,35,40,45 and 50 mg/kg; compound 1 group, 600,800,1000,1100,1200,1300, and 1400mg/kg, with about 20% of pexiganan) were given intraperitoneally, in a single administration, and death was noted daily for one week after administration. Adopting originPro 8 software to carry out data analysis and calculating respective LD of the positive drug and the tested drug50。
Pexiganan group: depending on the dose administered, the mice generally die within a half hour to 48 hours after administration and are substantially viable thereafter. After intraperitoneal injection, the mice in the high-dose group immediately showed extreme malaise state, curling, piloerection, and bluish eyes, limbs and tails until death. If the mice can recover from the bluish state to the original reddish state, the mice will not die. The low dose group animals appeared normal.
Table 5 pexiganan group mice dose grouping and mortality:
Figure BDA0000785761470000191
the data were analyzed using OriginPro 8 software and the median lethal dose LD50 was calculated to be 38.86 mg/kg.
Composite 1 group:
acute toxic reaction phenomenon: the same as positive control drug.
Table 6 dose grouping and mortality for compound 1 group mice:
Figure BDA0000785761470000192
data were analyzed using OriginPro 8 software and half the lethal dose LD50 was calculated to be 986.6 mg/kg. Calculated according to the actual dosage of pexiganan, LD50 is improved by 5 times.
Although specific embodiments of the invention have been described in detail, those skilled in the art will appreciate. Various modifications and substitutions of those details may be made in light of the overall teachings of the disclosure, and such changes are intended to be within the scope of the present invention. The full scope of the invention is given by the appended claims and any equivalents thereof.
Figure IDA0000785761560000011
Figure IDA0000785761560000021

Claims (13)

1. A complex comprising an antimicrobial peptide and a polymer, wherein the antimicrobial peptide is positively charged and the polymer is negatively charged;
the antibacterial peptide is pexiganan;
the polymer is polyethylene glycol-polyglutamic acid-poly isoleucine; and the number of the first and second electrodes,
the molar ratio of the polymer to the antibacterial peptide is 0.04: 1-15: 1.
2. The complex of claim 1, wherein the molar ratio of the polymer to the antimicrobial peptide is 0.2:1 to 5: 1.
3. The composite of claim 1, wherein the polymer is selected from the group consisting of:
polyethylene glycol5000-polyglutamic acid49-poly-isoleucine5；
Polyethylene glycol5000-polyglutamic acid49-poly-isoleucine13(ii) a And
polyethylene glycol2000-polyglutamic acid51-poly-isoleucine19。
4. A pharmaceutical composition comprising the complex of any one of claims 1-3, and a pharmaceutically acceptable carrier or excipient.
5. Use of a complex according to any one of claims 1 to 3 in the manufacture of a medicament for the prophylaxis or treatment of a disease for which an antimicrobial peptide is indicated.
6. The use of claim 5, wherein the disease to be prevented or treated by said antibacterial peptide is a disease caused by bacteria, fungi or viruses.
7. The use of claim 6, wherein the bacteria, fungi or viruses are bacteria, fungi or viruses that the antimicrobial peptide is capable of inhibiting or killing.
8. Use of a complex according to any one of claims 1 to 3 for inhibiting or killing bacteria, fungi or viruses in vitro for non-therapeutic purposes.
9. The use of claim 8, wherein the bacterium is a gram-positive or gram-negative bacterium.
10. The use of claim 8, wherein the bacteria, fungi or virus is a bacteria, fungi or virus that the antimicrobial peptide is capable of inhibiting or killing.
11. A method for preparing the complex of claim 1, comprising the steps of:
and respectively dissolving the antibacterial peptide and the polymer in water according to a certain proportion, stirring to fully mix the antibacterial peptide and the polymer, and separating to obtain the compound, wherein the molar ratio of the polymer to the antibacterial peptide is 0.04: 1-15: 1.
12. The preparation method of claim 11, wherein the molar ratio of the polymer to the antimicrobial peptide is 0.2:1 to 5: 1.
13. The preparation method of claim 11 or 12, further comprising a step of lyophilization after the separation.
CN201510518175.6A 2015-08-21 2015-08-21 Composite formed by combining antibacterial peptide and polymer, preparation method and application thereof Active CN106466475B (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
CN201510518175.6A CN106466475B (en) 2015-08-21 2015-08-21 Composite formed by combining antibacterial peptide and polymer, preparation method and application thereof
PCT/CN2016/095471 WO2017032236A1 (en) 2015-08-21 2016-08-16 Complex prepared from antimicrobial peptides in combination with polymers, and preparation method and use thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201510518175.6A CN106466475B (en) 2015-08-21 2015-08-21 Composite formed by combining antibacterial peptide and polymer, preparation method and application thereof

Publications (2)

Publication Number Publication Date
CN106466475A CN106466475A (en) 2017-03-01
CN106466475B true CN106466475B (en) 2021-06-08

Family

ID=58099612

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201510518175.6A Active CN106466475B (en) 2015-08-21 2015-08-21 Composite formed by combining antibacterial peptide and polymer, preparation method and application thereof

Country Status (2)

Country Link
CN (1) CN106466475B (en)
WO (1) WO2017032236A1 (en)

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108484904B (en) * 2018-04-20 2020-03-17 电子科技大学 temperature/pH dual-sensitive polypeptide random copolymer, and synthesis method and hydrogel system thereof
CN110092814B (en) * 2019-04-02 2022-08-09 同济大学 Amphiphilic multi-block antibacterial peptide and preparation method and application thereof
CN111110856B (en) * 2020-02-28 2022-12-16 苏州大学 Antibacterial drug based on antibacterial peptide and hydrophilic polymer and preparation method thereof
CN116744974A (en) * 2021-01-01 2023-09-12 普渡研究基金会 Co-formulation of polymyxin for inhalation
CN112920279B (en) * 2021-03-09 2022-07-05 海南大学 Anti-biological fouling type polymer peptide hydrogel material for extracting uranium from seawater and preparation method and application thereof
CN113087759B (en) * 2021-06-09 2021-08-27 南京杰肽生物科技有限公司 Synthetic method of antibacterial polypeptide derivative

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102657843A (en) * 2012-06-05 2012-09-12 云南民族大学 Antibacterial peptide composition and preparation method thereof
CN102665686A (en) * 2009-09-09 2012-09-12 药明公司 Anionic-core composition for delivery of therapeutic agents, and methods of making and using the same

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2012510798A (en) * 2008-12-05 2012-05-17 アンジオケム インコーポレーテッド Peptide therapeutic agent conjugates and uses thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102665686A (en) * 2009-09-09 2012-09-12 药明公司 Anionic-core composition for delivery of therapeutic agents, and methods of making and using the same
CN102657843A (en) * 2012-06-05 2012-09-12 云南民族大学 Antibacterial peptide composition and preparation method thereof

Also Published As

Publication number Publication date
CN106466475A (en) 2017-03-01
WO2017032236A1 (en) 2017-03-02

Similar Documents

Publication Publication Date Title
CN106466475B (en) Composite formed by combining antibacterial peptide and polymer, preparation method and application thereof
Liang et al. Role and modulation of the secondary structure of antimicrobial peptides to improve selectivity
US20200288709A1 (en) Composition with high antimicrobial activity and low toxicity
Cano-Garrido et al. Functional protein-based nanomaterial produced in microorganisms recognized as safe: a new platform for biotechnology
US8377871B2 (en) Dendrimeric peptides, pharmaceutical compositions and methods of using the same
JP6488129B2 (en) Cationic peptide polysaccharides with excellent broad spectrum antimicrobial activity and high selectivity
WO2018081845A1 (en) Antibacterial compositions and methods
KR101666548B1 (en) Bacterial extract for digestive or urinary tract disorders and process for its preparation
Grogg et al. Cell penetration, herbicidal activity, and in‐vivo‐toxicity of oligo‐arginine derivatives and of novel guanidinium‐rich compounds derived from the biopolymer cyanophycin
Lee et al. Cell-selectivity of tryptophan and tyrosine in amphiphilic α-helical antimicrobial peptides against drug-resistant bacteria
Roque-Borda et al. HPMCAS-Coated Alginate Microparticles Loaded with Ctx (Ile21)-Ha as a Promising Antimicrobial Agent against Salmonella Enteritidis in a Chicken Infection Model
US9309298B2 (en) Anti-bacterial polypeptides and pathogen specific synthetic antibodies
KR101983679B1 (en) Alpha-helical peptide having antimicrobial actvity against drug-resistant bacteria and biofilm and antimicrobial composition comprising the same
CN109641938A (en) Derived from Arginine-rich structural domain through modified antimicrobial peptide
JPH04502904A (en) Methods of administering amphipathic peptides and compositions thereof
JP2020528426A (en) Lactofericin and lactoferrampin for the treatment of infectious diseases
EP2152241A1 (en) Method of delivering a protein into a cell
Enright Design, Synthesis and Biological Evaluation of Biocompatible Cationic Graft Polymers as Antibacterial Agents
Li et al. Enzyme-Triggered Polyelectrolyte Complex for Responsive Delivery of α-Helical Polypeptides to Optimize Antibacterial Therapy
Reimer Characterization of outer membrane vesicle production and composition by cationic antimicrobial-adapted Escherichia coli
EP4199907A1 (en) Targeted antigen delivery system and uses thereof
JP2024510908A (en) Carrier proteins for peptide antigens
KR20190056606A (en) Composition for Drug Delivery into Nucleus Comprising Cytidine 5-triphosphate or Polymer thereof, and Immunostimulating Agents Comprising the Same
JPH11286452A (en) Agent for protecting infection of pathogenic bacterium and virus

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
GR01 Patent grant
GR01 Patent grant