Antimicrobial peptides and methods of making and using same
AssigneeThe Johns Hopkins University
InventorsCharles H. Chen, Martin B. Ulmschneider
Life of this patent
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Abstract
The present invention provides a new methodology combining MD simulations and database-guided high-throughput screening to rationally design pore forming membrane-active peptides. The present inventive methodology is able to allow tuning of a range of structural and functional properties such as pore size and selectively targeting membranes with specific lipid compositions. The present inventive methods will ultimately allow de novo design of membrane-active peptides for a wide range of biomedical applications, including for example, antimicrobial agents.
Claims10
A method for designing pore-forming, membrane-active peptides comprising the steps of:
- a) providing a first template membrane-active peptide;
- b) comparing the amino acid sequence of the first template peptide to the amino acid sequences of known membrane-active peptides located in a first membrane-active peptide database;
- c) identifying similarities in hydrophobicity and sequence composition between the first template peptide and the known membrane-active peptides;
- d) identifying amino acid locations in the first template peptide, which can be modified using the information from c);
- e) generating a plurality of modified membrane-active peptide sequences using the information from d);
- f) performing MD simulations on the plurality of modified membrane-active peptide sequences and determining whether the plurality of modified membrane-active peptide sequences have the physical characteristics of self-assembly into a pore-forming structure;
- g) synthesizing the plurality of modified membrane-active peptide sequences of e);
- h) testing the plurality of modified membrane-active peptides in an in vitro cell membrane testing system to determine efficacy of pore formation in cell membranes;
- and
- i) identifying one or more modified membrane-active peptides when the membrane testing system gives a positive result.
A pore-forming, membrane-active peptide having the amino acid sequence GLLDLLKLLLKAAG-COOH (SEQ ID NO:
- 3), GLLDLLHLLLKAAGW-AMIDE (SEQ ID NO:
- 4), GLADLAKLLLKLLGW-AMIDE (SEQ ID NO:
- 5), GLLDLLKLLLKLAGW-AMIDE (SEQ ID NO:
- 6), GLDDLAKLLLKLAGW-AMIDE (SEQ ID NO:
- 7), GLDDLLKALLKAAGW-AMIDE (SEQ ID NO:
- 8), GLLDDAKLLAKLAGW-AMIDE (SEQ ID NO:
- 9), GLLDLPKALAKALGW-AMIDE (SEQ ID NO:
- 10), GLADAAKLLLKAAGW-AMIDE (SEQ ID NO:
- 11), and GLLDAAKLLAKAAGW-AMIDE (SEQ ID NO:
- 12) and a functional homolog of or functional fragment thereof;
- and a fusion polypeptide comprising an amino acid sequence of any of the above peptides.
A pore-forming, membrane-active peptide selected from the group consisting of GLLDLLKLLLKAAG-COOH (SEQ ID NO:
- 3), GLLDLLHLLLKAAGW-AMIDE (SEQ ID NO:
- 4), GLADLAKLLLKLLGW-AMIDE (SEQ ID NO:
- 5), GLLDLLKLLLKLAGW-AMIDE (SEQ ID NO:
- 6), GLDDLAKLLLKLAGW-AMIDE (SEQ ID NO:
- 7), GLDDLLKALLKAAGW-AMIDE (SEQ ID NO:
- 8), GLLDDAKLLAKLAGW-AMIDE (SEQ ID NO:
- 9), GLLDLPKALAKALGW-AMIDE (SEQ ID NO:
- 10), GLADAAKLLLKAAGW-AMIDE (SEQ ID NO:
- 11), and GLLDAAKLLAKAAGW-AMIDE (SEQ ID NO:
- 12) or a functional homolog of or functional fragment thereof, or a fusion polypeptide comprising an amino acid sequence of any of the above peptides, in an effective amount, for use in the treatment of a microbial infection in a subject in need thereof.
A pharmaceutical composition comprising one or more antimicrobial peptides selected from the group consisting of GLLDLLKLLLKAAG-COOH (SEQ ID NO:
- 3), GLLDLLHLLLKAAGW-AMIDE (SEQ ID NO:
- 4), GLADLAKLLLKLLGW-AMIDE (SEQ ID NO:
- 5), GLLDLLKLLLKLAGW-AMIDE (SEQ ID NO:
- 6), GLDDLAKLLLKLAGW-AMIDE (SEQ ID NO:
- 7), GLDDLLKALLKAAGW-AMIDE (SEQ ID NO:
- 8), GLLDDAKLLAKLAGW-AMIDE (SEQ ID NO:
- 9), GLLDLPKALAKALGW-AMIDE (SEQ ID NO:
- 10), GLADAAKLLLKAAGW-AMIDE (SEQ ID NO:
- 11), and GLLDAAKLLAKAAGW-AMIDE (SEQ ID NO:
- 12) or a functional homolog of or functional fragment thereof;
- or a fusion polypeptide comprising an amino acid sequence of any of the above peptides, and a pharmaceutically acceptable carrier in an effective amount, for use in the treatment of a microbial infection in a subject in need thereof.
A pharmaceutical composition comprising one or more antimicrobial peptides selected from the group consisting of GLLDLLKLLLKAAG-COOH (SEQ ID NO:
- 3), GLLDLLHLLLKAAGW-AMIDE (SEQ ID NO:
- 4), GLADLAKLLLKLLGW-AMIDE (SEQ ID NO:
- 5), GLLDLLKLLLKLAGW-AMIDE (SEQ ID NO:
- 6), GLDDLAKLLLKLAGW-AMIDE (SEQ ID NO:
- 7), GLDDLLKALLKAAGW-AMIDE (SEQ ID NO:
- 8), GLLDDAKLLAKLAGW-AMIDE (SEQ ID NO:
- 9), GLLDLPKALAKALGW-AMIDE (SEQ ID NO:
- 10), GLADAAKLLLKAAGW-AMIDE (SEQ ID NO:
- 11), and GLLDAAKLLAKAAGW-AMIDE (SEQ ID NO:
- 12) or a functional homolog of or functional fragment thereof;
- or a fusion polypeptide comprising an amino acid sequence of any of the above peptides, at least one additional biologically active agent, and a pharmaceutically acceptable carrier, in an effective amount, for use in the treatment of a microbial infection in a subject in need thereof.
A method of treating a surface to prevent or remove microbial growth comprising applying to the surface an effective amount of one or more antimicrobial peptides selected from the group consisting of GLLDLLKLLLKAAG-COOH (SEQ ID NO:
- 3), GLLDLLHLLLKAAGW-AMIDE (SEQ ID NO:
- 4), GLADLAKLLLKLLGW-AMIDE (SEQ ID NO:
- 5), GLLDLLKLLLKLAGW-AMIDE (SEQ ID NO:
- 6), GLDDLAKLLLKLAGW-AMIDE (SEQ ID NO:
- 7), GLDDLLKALLKAAGW-AMIDE (SEQ ID NO:
- 8), GLLDDAKLLAKLAGW-AMIDE (SEQ ID NO:
- 9), GLLDLPKALAKALGW-AMIDE (SEQ ID NO:
- 10), GLADAAKLLLKAAGW-AMIDE (SEQ ID NO:
- 11), and GLLDAAKLLAKAAGW-AMIDE (SEQ ID NO:
- 12) or a functional homolog of or functional fragment thereof;
- or a fusion polypeptide comprising an amino acid sequence of any of the above peptides.
A pore-forming, membrane-active peptide having the amino acid sequence of GxxD4xxK7xxxK11xxGW-Amide, where ‘x’ is independently any one of the four amino acids (Ala, Leu, Lys, and Asp) with the proviso that the sequence cannot be SEQ ID NO:
- 22.
Dependencies are inferred from the claim text.