Stabilized compounds having secondary structure motifs

Patent metadata

Field	Value
Publication number	US-10487110-B2
Application number	US-201815917560-A
Country	US
Kind code	B2
Filing date	Mar 9, 2018
Priority date	May 18, 1999
Publication date	Nov 26, 2019
Grant date	Nov 26, 2019

Abstract

The present invention provides novel stabilized crosslinked compounds having secondary structure motifs, libraries of these novel compounds, and methods for the synthesis of these compounds libraries thereof. The synthesis of these novel stabilized compounds involves (1) synthesizing a peptide from a selected number of natural or non-natural amino acids, wherein the peptide comprises at least two moieties capable of undergoing reaction to promote carbon-carbon bond formation; and (2) contacting the peptide with a reagent to generate at least one crosslinker and to effect stabilization of a secondary structure motif. The present invention, in a preferred embodiment, provides stabilized p53 donor helical peptides. Additionally, the present invention provides methods for disrupting the p53/MDM2 binding interaction comprising (1) providing a crosslinked stabilized α-helical structure; and (2) contacting the crosslinked stabilized α-helical structure with MDM2.

First claim

We claim:

1. A method for disrupting the binding of a first protein to a second protein, the method comprising contacting a peptide with the second protein, wherein the first protein is a native protein; the second protein is another native protein; the second protein comprises a helix acceptor; the first protein binds the helix acceptor of the second protein; the peptide binds the helix acceptor of the second protein; the peptide comprises a stabilized alpha helix; and the peptide comprises at least two amino acids connected by a hydrocarbon covalent cross-link.

2. The method of claim 1 , wherein the hydrocarbon covalent cross-link stabilizes the alpha-helix of the peptide.

3. The method of claim 1 , wherein the peptide comprises a binding site of a donor helix of the first protein.

4. The method of claim 1 , wherein the cross-link comprises a carbon-carbon double bond.

5. The method of claim 1 , wherein the at least two amino acids connected by the hydrocarbon covalent cross-link are positioned at i,i+4 or at i,i+7.

6. The method of claim 1 , wherein the at least two amino acids that are connected by the hydrocarbon covalent cross-link are alpha,alpha-methyl, vinyl; alpha, alpha-methyl, alkylolefin; or divinyl amino acids.

7. The method of claim 6 , wherein the at least two amino acids that are connected by the hydrocarbon covalent cross-link are positioned at i,i+4 or at i,i+7.

8. The method of claim 1 , wherein at least one amino acid that is connected by the hydrocarbon covalent cross-link comprises an alpha-methyl, alpha-alkenyl amino acid.

9. The method of claim 1 , wherein the hydrocarbon covalent cross-link spans from one to two turns on the alpha-helix.

10. A method for synthesizing a peptide comprising a cross-link, the method comprising:
(a) synthesizing an amino acid sequence, wherein the amino acid sequence comprises a first amino acid comprising a first moiety and a second amino acid comprising a second moiety, and wherein the first and second moieties are reactive toward one another in the presence of a catalyst; and
(b) reacting the amino acid sequence under conditions sufficient to promote a reaction between the first and second moieties toward each other in the presence of the catalyst, thereby resulting in formation of the cross-link in the peptide; wherein the peptide comprises a stabilized alpha helix and can disrupt binding of a first protein to a second protein; wherein the first protein is a native protein; the second protein is another native protein; the second protein comprises a helix acceptor; and the first protein binds the helix acceptor of the second protein.

11. The method of claim 10 , wherein the cross-link spans from one to two turns on the alpha-helix.

12. The method of claim 10 , wherein the cross-link comprises a carbon-carbon double bond.

13. The method of claim 10 , wherein the peptide is stabilized in comparison to a corresponding uncross-linked peptide.

14. The method of claim 10 , wherein the second protein is a natural protein.

15. The method of claim 10 , wherein the peptide comprises a binding site of a donor helix of the first protein.

16. The method of claim 10 , wherein at least one amino acid in the amino acid sequence is an alpha-allyl amino acid.

17. The method of claim 10 , wherein the cross-link stabilizes an alpha-helix of the peptide.

18. The method of claim 1 , wherein the peptide is stabilized in comparison to a corresponding uncross-linked peptide.

Assignees

• Harvard College

Inventors

• Verdine Gregory L
• Schafmeister Christian E

Classifications

• C07K1/10

  using coupling agents {(C07K1/006 takes precedence)} · CPC title

• C07K14/4746

  p53 · CPC title

• C07K1/00 Primary

  General methods for the preparation of peptides {, i.e. processes for the organic chemical preparation of peptides or proteins of any length} · CPC title

• C07K1/047 Primary

  Simultaneous synthesis of different peptide species; Peptide libraries · CPC title

• C07K1/113

  without change of the primary structure · CPC title