Antimicrobial peptides with reduced hemolysis and methods of their use

• US 20060128614A1
• Filed: 12/02/2004
• Published: 06/15/2006
• Est. Priority Date: 09/20/2002
• Status: Abandoned Application

• Current Assignee: Pacgen Biopharmaceuticals Corporation (General Biologicals Corporation)
• Original Assignee: Pacgen Biopharmaceuticals Corporation (General Biologicals Corporation)
• Inventors: Cheng, Jya-Wei, Tzeng, Shiou-Ru
• Application Number:
  US11/000,970
• Publication Number:
  US 20060128614A1
• Time in Patent Office: Days
• US Class Current: 514/2.100
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: C07K 5/1019 with the first amino acid being basic
• CPC Class Codes: C07K 7/02 Linear peptides containing at least one abnormal peptide link
• CPC Class Codes: C07K 7/06 having 5 to 11 amino acids
• CPC Class Codes: C07K 7/64 Cyclic peptides containing only normal peptide links

Abstract

The present invention provides novel cyclic and linear short peptides containing one of the following amino acid residue sequences: Xa₁-Naa-Xa₁-Xa₁-Naa-Xa₂ or Xa₁-Naa-Xa₂-Xa₁-Naa-Xa₁ wherein: Xa₁ represents lysine, arginine, or histidine; Naa represents an unnatural hydrophobic aromatic amino acid moiety selected from the group consisting of (naphtha-1-yl)alanine (1-Nal), (naphtha-2-yl)alanine (2-Nal), (benzothien-3-yl)alanine (Bal), diphenylalanine (Dip), (4,4′-biphen-yl)alanine (Bip), (anthracen-9-yl)alanine (Ath), and (2,5,7-tri-tert-butyl-indol-3-yl)alanine (Tht); and Xa₂ represents valine, leucine, or isoleucine. The novel peptides exhibit broad spectrum antimicrobial activity against Gram-positive and Gram-negative bacteria and fungi by effecting modification of the primary peptide structure. Further, the peptides exhibit improved serum compatibility and reduced hemolysis.

Claims

1. The group of isolated and purified cyclic or linear peptides, each with microbicidal activity, described by one of the following formulae:

Xa₁-Naa-Xa₁-Xa₁-Naa-Xa₂

or

Xa₁-Naa-Xa₂-Xa₁-Naa-Xa₁

wherein;

Xa₁ represents a hydrophilic basic amino acid moiety selected from the group consisting of lysine, arginine, and histidine;

Naa represents an unnatural hydrophobic aromatic amino acid moiety selected from the group consisting of (naphtha-1-yl)alanine (1-Nal), (naphtha-2-yl)alanine (2-Nal), (benzothien-3-yl)alanine (Bal), diphenylalanine (Dip), (4,4′

-biphen-yl)alanine (Bip), (anthracen-9-yl)alanine (Ath), and (2,5,7-tri-tert-butyl-indol-3-yl)alanine (Tht); and

Xa₂ represents a hydrophobic amino acid moiety selected from the group consisting of valine, leucine, and isoleucine.

2. An isolated and purified peptide chosen from the group of peptides of claim 1, which is selected from the group consisting of

3. The isolated and purified peptides of claim 1, wherein the isolated and purified peptides are of linear topology.

4. The isolated and purified peptides of claim 1, wherein the isolated and purified peptides are of cyclic topology.

5. An isolated and purified peptide of claim 1, wherein the isolated and purified peptide is amidated at the C-terminal amino acid.

6. An isolated and purified peptide of claim 1, wherein the peptide is acetylated at the N-terminal amino acid.

7. An isolated and purified peptide of claim 1, wherein the peptide is esterified at the C-terminal amino acid.

8. An isolated and purified peptide of claim 1 which has at least one amino acid altered to a corresponding D-amino acid.

9. A composition comprising an isolated and purified peptide according to claim 1 in a mixture with a carrier or excipient.

10. A pharmaceutical composition which comprises a peptide of claim 1 and a pharmaceutical carrier.

11. A peptide which is the retro-oriented amino acid sequence of a peptide of claim 1.

12. A method to inactivate an endotoxin of Gram-negative bacteria comprising the step of administering an effective amount of an isolated and purified antimicrobial peptide according to claim 1 to a subject in need thereof.

13. A method to treat a microbial or viral infection in a subject comprising the step of administering an effective amount of an isolated and purified antimicrobial peptide according to claim 1 to the subject.

14. A method to inhibit the growth of a microbe comprising the step of administering an effective amount of an isolated and purified antimicrobial peptide according to claim 1 to a subject in need thereof.

15. A peptide of claim 1 which is of the formula Lys-(2-Nal)-Arg-Arg-(2-Nal)-Val-Arg-(2-Nal)-Ile.

16. A pharmaceutical composition which comprises a peptide of claim 15 and a pharmaceutical carrier.