Stabilized anti-microbial peptides for the treatment of antibiotic-resistant bacterial infections
Methods of generating and optimizing stabilized (e.g., stapled and/or stitched) anti-microbial peptides (StAMPs) for the prophylaxis and treatment of antibiotic-resistant (e.g., colistin-resistant, methicillin resistant, meropenem-resistant) bacterial infections (e.g., Gram-negative, Gram-positive), and methods for using such peptides for experimental investigation, livestock management, management of crops/trees/plants, and/or therapeutic benefit. Also featured are methods for reducing renal toxicity of a StAMP.
1. A stapled peptide or pharmaceutically acceptable salt thereof comprising the amino acid sequence: GX 1 GKFX 2 HSKKKFGKAX 3 VGEX 4 AKK (SEQ ID NO:43), wherein each of X 1 , X 2 , X 3 , and X 4 is a non-natural amino acid; wherein X 1 and X 2 are cross-linked to each other; and wherein X 3 and X 4 are cross-linked to each other.
2. The stapled peptide or pharmaceutically acceptable salt thereof of claim 1 , wherein each of X 1 , X 2 , X 3 , and X 4 is independently an α, α-disubstituted non-natural amino acid comprising an olefinic side chain.
3. The stapled peptide or pharmaceutically acceptable salt thereof of claim 1 , wherein each of X 1 , X 2 , X 3 , and X 4 is (S)-2-(4-pentenyl)Ala-OH.
4. A pharmaceutical composition comprising the stapled peptide or pharmaceutically acceptable salt thereof of claim 3 , and a pharmaceutically acceptable carrier.
5. The stapled peptide or pharmaceutically acceptable salt thereof of claim 1 , wherein the stapled peptide is between 23 and 30 amino acids in length, inclusive.
6. A pharmaceutical composition comprising the stapled peptide or pharmaceutically acceptable salt thereof of claim 1 , and a pharmaceutically acceptable carrier.
7. A stapled peptide or pharmaceutically acceptable salt thereof consisting of the amino acid sequence: GX 1 GKFX 2 HSKKKFGKAX 3 VGEX 4 AKK (SEQ ID NO:43), wherein each of X 1 , X 2 , X 3 , and X 4 is (S)-2-(4-pentenyl)Ala-OH; wherein X 1 and X 2 are cross-linked to each other; and wherein X 3 and X 4 are cross-linked to each other.
8. The stapled peptide or pharmaceutically acceptable salt thereof of claim 1 , wherein the stapled peptide is modified by acetylation, PEGylation, amidation, and/or modification with a fatty acid chain.
9. A pharmaceutical composition comprising the stapled peptide or pharmaceutically acceptable salt thereof of claim 7 , and a pharmaceutically acceptable carrier.
10. A method of treating a bacterial infection in a subject in need thereof, the method comprising: administering a therapeutically effective amount of the stapled peptide or pharmaceutically acceptable salt thereof of claim 1 to the subject.
11. The method of claim 10 , wherein the bacterial infection is a Gram-positive bacterial infection.
12. The method of claim 10 , wherein the bacterial infection is a Gram-negative bacterial infection.
13. The method of claim 10 , wherein the bacterial infection is an E. coli infection, a K. pneumonia infection, a Salmonella infection, an Enterobacter cloacae infection, a Shigella infection, a B. cereus infection, a P. aeruginosa infection, a S. aureus infection, or an A. baumannii infection.
14. The method of claim 10 , wherein the bacterial infection is an antibiotic-resistant bacterial infection.
15. The method of claim 14 , wherein the antibiotic-resistant bacterial infection is a Gram-negative colistin-resistant bacterial infection or a Gram-positive carbapenem-resistant bacterial infection.
16. The method of claim 14 , wherein the antibiotic-resistant bacterial infection is an Actinomyces israelii infection, an Actinomyces gerencseriae infection, a Propionibacterium propionicus infection, a Bacillus anthracis infection, an Arcanobacterium haemolyticum infection, a Bacillus cereus infection, a bacterial vaginosis microbiota infection, a Clostridium botulinum infection, a Mycobacterium ulcerans infection, a Group A Streptococcus or Staphylococcus infection, a Clostridium species infection, a Corynebacterium diphtheria infection, an Enterococcus species infection, a Streptococcus pyogenes infection, a Streptococcus agalactiae infection, a Mycobacterium leprae infection, a Mycobacterium lepromatosis infection, a Listeria monocytogenes infection, a Nocardia species infection, a Streptococcus pneumonia infection, a Staphylococcus species infection, a Mycobacterium tuberculosis infection, a Yersinia pseudotuberculosis infection, or a Yersinia enterocolitica infection.
17. The method of claim 14 , wherein the antibiotic-resistant bacterial infection is a Pseudomonas aeruginosa infection, a Neisseria gonorrhoeae infection, a Chlamydia trachomatis infection, a Yersinia pestis infection, a Proteus mirabilis infection, an Enterobacter cloacae infection, a Serratia marcescens infection, a Helicobacter pylori infection, a Salmonella enteritidis infection, a Salmonella typhi infection, a Shigella infection, a Moraxella infection, a Stenotrophomonas infection, a Bdellovibrio infection, an acetic acid bacterial infection, a Klebsiella pneumoniae infection, a Legionella pneumophila infection, an A. baumannii infection, an E. coli infection, or an Enterobacter infection.
18. The method of claim 10 , wherein the subject is a human.
19. The method of claim 10 , wherein the subject is an animal.
20. A method of killing or inhibiting the growth of a bacterium, the method comprising contacting the bacterium with the stapled peptide or pharmaceutically acceptable salt thereof of claim 1 .
21. The method of claim 20 , wherein the bacterium is an antibiotic-resistant bacterium.
22. The method of claim 20 , wherein the contacting is in vitro.
23. The method of claim 20 , wherein the contacting is in vivo.
24. A method of treating a bacterial infection in a subject in need thereof, the method comprising: administering a therapeutically effective amount of the stapled peptide or pharmaceutically acceptable salt thereof of claim 7 to the subject.
25. A method of making a stapled peptide or pharmaceutically acceptable salt thereof, the method comprising:
(a) providing a peptide comprising the amino acid sequence: GX 1 GKFX 2 HSKKKFGKAX 3 VGEX 4 AKK (SEQ ID NO:43), or pharmaceutically acceptable salt thereof, wherein each of X 1 , X 2 , X 3 , and X 4 is an α, α-disubstituted non-natural amino acid comprising an olefinic side chain; and
(b) performing a ring-closing metathesis reaction on the peptide, thereby making the stapled peptide or pharmaceutically acceptable salt thereof.
26. The method of claim 25 , further comprising formulating the stapled peptide or pharmaceutically acceptable salt thereof as a sterile pharmaceutical composition.
27. The method of claim 24 , wherein the subject is an animal.
28. The method of claim 27 , wherein the subject is a human.
29. A method of killing or inhibiting the growth of a bacterium, the method comprising contacting the bacterium with the stapled peptide or pharmaceutically acceptable salt thereof of claim 7 .
30. The method of claim 29 , wherein the subject is a human.
31. The method of claim 29 , wherein the subject is an animal.