Stapled intracellular-targeting antimicrobial peptides to treat infection

Patent metadata

Field	Value
Publication number	US-11945846-B2
Application number	US-202016813528-A
Country	US
Kind code	B2
Filing date	Mar 9, 2020
Priority date	Feb 29, 2016
Publication date	Apr 2, 2024
Grant date	Apr 2, 2024

Abstract

Structurally stabilized, e.g., stapled, peptides with the ability to translocate through microbial cell membranes to the interior of microbial cells and exert a biological activity there are provided, as are methods of designing, making and using such peptides.

First claim

What is claimed is:

1. A stapled peptide or a pharmaceutically acceptable salt thereof comprising the amino acid sequence TRSSRAGLQWPVGRVX 1 RLLX 2 K (SEQ ID NO:38), TRSSRAGLQWPX 1 GRVX 2 RLLRK (SEQ ID NO:42), TRSSRAGLX 1 WPVX 2 RVHRLLRK (SEQ ID NO:43), TRX 1 SRAX 2 LQWPVGRVHRLLRK (SEQ ID NO:47), TRSSRAX 3 LQWPVGX 4 VHRLLRK (SEQ ID NO:55), or TRSSX 3 AGLQWPX 4 GRVHRLLRK (SEQ ID NO:57),
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 where present, and
wherein X 3 and X 4 are cross-linked to each other where present.

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 and X 2 , and X 4 is (S)-2-(4′-pentenyl)alanine, and wherein X 3 is (R)-2-(7′-octenyl)alanine.

4. The stapled peptide or pharmaceutically acceptable salt thereof of claim 1 , comprising the amino acid sequence TRSSRAGLX 1 WPVX 2 RVHRLLRK (SEQ ID NO:43), TRX 1 SRAX 2 LQWPVGRVHRLLRK (SEQ ID NO:47), or TRSSRAX 3 LQWPVGX 4 VHRLLRK (SEQ ID NO:55), wherein each of X 1 and X 2 , and X 4 is (S)-2-(4′-pentenyl)alanine, and wherein X 3 is (R)-2-(7′-octenyl)alanine.

5. The stapled peptide or pharmaceutically acceptable salt thereof of claim 1 , which is 21 to 30 amino acids in length.

6. The stapled peptide or pharmaceutically acceptable salt thereof of claim 1 , wherein the pharmaceutically acceptable salt is an acetate, a citrate, a fumarate, a maleate, a succinate, a sulfate, or a malonate.

7. The stapled peptide or pharmaceutically acceptable salt thereof of claim 1 , consisting of the amino acid sequence
TRSSRAGLQWPVGRVX 1 RLLX 2 K (SEQ ID NO:38), TRSSRAGLQWPX 1 GRVX 2 RLLRK (SEQ ID NO:42), TRSSRAGLX 1 WPVX 2 RVHRLLRK (SEQ ID NO:43), TRX 1 SRAX 2 LQWPVGRVHRLLRK (SEQ ID NO:47), TRSSRAX 3 LQWPVGX 4 VHRLLRK (SEQ ID NO:55), or TRSSX 3 AGLQWPX 4 GRVHRLLRK (SEQ ID NO:57).

8. The stapled peptide or pharmaceutically acceptable salt thereof of claim 7 , 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.

9. The stapled peptide or pharmaceutically acceptable salt thereof of claim 7 , wherein each of X 1 and X 2 , and X 4 is (S)-2-(4′-pentenyl)alanine, and wherein X 3 is (R)-2-(7′-octenyl)alanine.

10. The stapled peptide or pharmaceutically acceptable salt thereof of claim 1 , consisting of the amino acid sequence TRSSRAGLX 1 WPVX 2 RVHRLLRK (SEQ ID NO:43), TRX 1 SRAX 2 LQWPVGRVHRLLRK (SEQ ID NO:47), or TRSSRAX 3 LQWPVGX 4 VHRLLRK (SEQ ID NO:55), wherein each of X 1 and X 2 , and X 4 is (S)-2-(4′-pentenyl)alanine, and wherein X 3 is (R)-2-(7′-octenyl)alanine.

11. The stapled peptide or pharmaceutically acceptable salt thereof of claim 1 , wherein the stapled peptide or pharmaceutically acceptable salt thereof is dihydroxylated.

12. The stapled peptide or pharmaceutically acceptable salt thereof of claim 1 , which is linked to an antibiotic or methotrexate.

13. The stapled peptide or pharmaceutically acceptable salt thereof of claim 12 , wherein the antibiotic is a Gram-positive antimicrobial.

14. The stapled peptide or pharmaceutically acceptable salt thereof of claim 13 , wherein the Gram-positive antimicrobial is a beta-lactam drug.

15. A pharmaceutical composition comprising the stapled peptide or pharmaceutically acceptable salt thereof of claim 1 , and a pharmaceutically acceptable carrier.

16. A pharmaceutical composition comprising the stapled peptide or pharmaceutically acceptable salt thereof of claim 3 , and a pharmaceutically acceptable carrier.

17. A pharmaceutical composition comprising the stapled peptide or pharmaceutically acceptable salt thereof of claim 4 , and a pharmaceutically acceptable carrier.

18. A pharmaceutical composition comprising the stapled peptide or pharmaceutically acceptable salt thereof of claim 7 , and a pharmaceutically acceptable carrier.

19. A pharmaceutical composition comprising the stapled peptide or pharmaceutically acceptable salt thereof of claim 10 , and a pharmaceutically acceptable carrier.

20. A method of treating a bacterial infection in a human 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 human subject.

21. The method of claim 20 , wherein the bacterial infection is caused by a Gram-positive bacterium.

22. The method of claim 20 , wherein the bacterial infection is caused by a Gram-negative bacterium.

23. The method of claim 20 , wherein the bacterial infection is caused by E. coli, B. cereus, P aeruginosa, S. aureus , or methicillin-resistant S. aureus.

24. The method of claim 20 , wherein the stapled peptide or pharmaceutically acceptable salt thereof comprises the amino acid sequence TRSSRAGLX 1 WPVX 2 RVHRLLRK (SEQ ID NO:43), TRX 1 SRAX 2 LQWPVGRVHRLLRK (SEQ ID NO:47), or TRSSRAX 3 LQWPVGX 4 VHRLLRK (SEQ ID NO:55), wherein each of X 1 and X 2 , and X 4 is (S)-2-(4′-pentenyl)alanine, and wherein X 3 is (R)-2-(7′-octenyl)alanine.

25. The method of claim 20 , further comprising administering to the subject a therapeutically-effective amount of an antibiotic.

26. A method of making a stapled peptide or pharmaceutically acceptable salt thereof, the method comprising:
(a) providing a peptide comprising the amino acid sequence:
TRSSRAGLQWPVGRVX 1 RLLX 2 K (SEQ ID NO:38), TRSSRAGLQWPX 1 GRVX 2 RLLRK (SEQ ID NO:42), TRSSRAGLX 1 WPVX 2 RVHRLLRK (SEQ ID NO:43), TRX 1 SRAX 2 LQWPVGRVHRLLRK (SEQ ID NO:47), TRSSRAX 3 LQWPVGX 4 VHRLLRK (SEQ ID NO:55), or TRSSX 3 AGLQWPX 4 GRVHRLLRK (SEQ ID NO:57), 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.

27. The method of claim 26 , further comprising formulating the stapled peptide as a sterile pharmaceutical composition.

Assignees

• Dana Farber Cancer Inst Inc

Inventors

• Walensky Loren D
• Mourtada Rida

Classifications

• C07K14/461 Primary

  from fish · CPC title

• A61K38/08 Primary

  Peptides having 5 to 11 amino acids {(A61K38/043 - A61K38/046 take precedence)} · CPC title

• A61K38/10

  Peptides having 12 to 20 amino acids {(A61K38/043 - A61K38/046 take precedence)} · CPC title

• A61K38/1703

  from vertebrates · CPC title

• A61K38/1706

  from fish · CPC title