Compositions and uses of materials with high antimicrobial activity and low toxicity

• US 9,017,730 B2
• Filed: 02/25/2013
• Issued: 04/28/2015
• Est. Priority Date: 08/23/2010
• Status: Active Grant

• Current Assignee: Amicrobe, Inc., Regents of the University of California (University of California)
• Original Assignee: Amicrobe, Inc., Regents of the University of California (University of California)
• Inventors: Bevilacqua, Michael P., Benitez, Diego, Deming, Timothy J., Hanson, Jarrod A., Koziol, Lucas
• Primary Examiner(s): Witz, Jean
• Assistant Examiner(s): NEWMAN, MINDY B
• Application Number:
  US13/776,221
• Publication Number:
  US 20130267458A1
• Time in Patent Office: 792 Days
• Field of Search: None
• US Class Current: 424/491
• CPC Class Codes: A01N 37/18 containing the group —CO—N<, e.g. carboxylic acid amides or imides; Thio analogues thereof
• CPC Class Codes: A23L 3/3526 Organic compounds containing nitrogen
• CPC Class Codes: A61K 38/02 Peptides of undefined number of amino acids; Derivatives thereof
• CPC Class Codes: A61K 38/16 Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof enzyme inhibitors A61K38/005
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61K 47/02 Inorganic compounds
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61K 9/107 Emulsions ; Emulsion preconcentrates; Micelles composition of emulsions A61K47/00
• CPC Class Codes: A61P 13/02 of urine or of the urinary tract, e.g. urine acidifiers
• CPC Class Codes: A61P 15/02 for disorders of the vagina
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/02 Local antiseptics
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/001 by chemical synthesis
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

Improved synthetic copolypeptide antimicrobials contain cationic amino acid residues and may be based on a blocky sequence. These antimicrobials show low mammalian toxicity and may undergo directed self-assembly. The inventive synthetic copolypeptides are useful in treatment of wounds and other infections.

Claims

1. An antimicrobial composition comprising:

at least one species of synthetic copolypeptide of at least forty amino acid residues comprising;

at least one hydrophilic segment containing at least five contiguous cationic amino acid residues; and

at least one hydrophobic segment containing at least five contiguous hydrophobic amino acid residues;

wherein the hydrophilic segment contains a larger number of amino acid residues than the hydrophobic segment; and

water;

wherein said at least one species of synthetic copolypeptide forms multimeric structures in aqueous media;

wherein said at least one species of synthetic copolypeptide inhibits or kills microbes in aqueous media; and

wherein the composition inhibits or kills microbes.

2. The composition as described in claim 1, wherein said at least one species of synthetic copolypeptide comprises substantially only natural amino acids.

3. The composition as described in claim 1, wherein the ratio of the number of amino acids in said hydrophilic segment to the number of amino acids in said hydrophobic segment is at least 1.8 to 1.

4. The composition as described in claim 1, wherein the structures formed in aqueous media are selected from the group consisting of multimers in solution, micelles, sheets, vesicles, and fibrils.

5. The composition as described in claim 1, wherein said at least one species of synthetic copolypeptide is characterized by the ability to form mixtures in water without visible precipitate at room temperature at concentrations up to 10 fold above the critical aggregation concentration (CAC).

6. The composition as described in claim 1, wherein said at least one species of synthetic copolypeptide is characterized by the ability to form mixtures in water without visible precipitate at room temperature at concentrations up to 100 fold above the critical aggregation concentration (CAC).

7. The composition as described in claim 1, wherein said at least one species of synthetic copolypeptide has a critical aggregation concentration (CAC) in water that is at least 1 log lower than that of a random sequence copolypeptide of the same amino acid composition.

8. The composition as described in claim 1 characterized in that it kills or inhibits microbes in vitro at a lower concentration than it kills mammalian cells in vitro.

9. The composition as described in claim 1 characterized in that it kills or inhibits microbes in or on mammalian tissues in vivo at concentrations that show low toxicity for those tissues.

10. The composition as described in claim 1 characterized in that said at least one species of synthetic copolypeptide forms mixtures in water without visible precipitate at room temperature at concentrations of at least 1000 μ

g/mL.

11. The composition as described in claim 1 characterized in that it kills or inhibits microbes in vitro as measured by greater than 3 logs killing of Staphylococcus epidermidis and Escherichia coli in standard 60 minute time-kill assays at concentrations of said at least one species of synthetic copolypeptide of 100 μ

g/mL or less.

12. The composition as described in claim 1 characterized in that said at least one species of synthetic copolypeptide forms mixtures in water without visible precipitate at room temperature at concentrations at least 10 fold above the concentration required to inhibit or kill microbes in vitro as measured by greater than 3 logs killing of Staphylococcus epidermidis and Escherichia coli in standard 60 minute time-kill assays.

13. The composition as described in claim 1 having a storage modulus of at least 50 Pa at a concentration of said at least one species of synthetic copolypeptide of less than 40 mg/mL.

14. The composition as described in claim 1, wherein said at least one species of synthetic copolypeptide promotes platelet aggregation.

15. The composition as described in claim 1, wherein said at least one species of synthetic copolypeptide inhibits fibrinolysis.

16. The composition as described in claim 1 further comprising a combination of immiscible phases in a dispersed mixture or emulsion.

17. The composition as described in claim 1 formulated as a solution, a gel, a cream, a foam, or a dressing.

18. The composition as described in claim 1, further comprising an added active pharmaceutical ingredient (API) selected from steroids, pro-inflammatory agents, anti-inflammatory agents, anti-acne agents, preservatives, hemostatic agents, angiogenic agents, wound healing agents, anti-cancer agents and other antimicrobial agents.

19. The use of the composition as described in claim 1 for prevention or treatment of infection, for topical anti-infection, for microbial decolonization, for wound treatment, for surgical site treatment, for trauma treatment, for burn treatment, for treatment of diabetic foot ulcers, for eye treatment, for vaginal infections, for urinary tract infections, for hand sanitization, for coating prosthetic devices and implants, for food preservation and for solution preservation.

20. A method for the prevention or treatment of infections in a mammal comprising the step of:

applying to a site an antimicrobial composition comprising;

at least one species of synthetic copolypeptide of at least forty amino acid residues comprising;

at least one hydrophilic segment containing at least five contiguous cationic amino acid residues; and

at least one hydrophobic segment containing at least five contiguous hydrophobic amino acid residues;

wherein the hydrophilic segment contains a larger number of amino acid residues than the hydrophobic segment; and

water;

wherein said at least one species of synthetic copolypeptide forms multimeric structures in aqueous media;

wherein said at least one species of synthetic copolypeptide inhibits or kills microbes in aqueous media; and

wherein the composition inhibits or kills microbes.