Stabilized compounds having secondary structure motifs

• US 9,505,801 B2
• Filed: 10/31/2013
• Issued: 11/29/2016
• Est. Priority Date: 05/18/1999
• Status: Expired due to Fees

• Current Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Original Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Inventors: Verdine, Gregory L., Schafmeister, Christian E.
• Primary Examiner(s): Russel, Jeffrey E
• Application Number:
  US14/068,844
• Publication Number:
  US 20140162339A1
• Time in Patent Office: 1,125 Days
• Field of Search: None
• US Class Current: 1/1
• 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: A61K 38/02 Peptides of undefined number of amino acids; Derivatives thereof
• CPC Class Codes: C07K 1/00 General methods for the preparation of peptides , i.e. processes for the organic chemical preparation of peptides or proteins of any length
• CPC Class Codes: C07K 1/04 on carriers C07K1/003, C07K1/006 take precedence
• CPC Class Codes: C07K 1/047 Simultaneous synthesis of different peptide species; Peptide libraries
• CPC Class Codes: C07K 1/10 using coupling agents C07K1/006 takes precedence
• CPC Class Codes: C07K 1/113 without change of the primary structure
• CPC Class Codes: C07K 14/001 by chemical synthesis
• CPC Class Codes: C07K 14/4746 p53
• CPC Class Codes: C12N 15/1093 General methods of preparing gene libraries, not provided for in other subgroups

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.

Claims

1. A method for disrupting the binding of a p53 protein to an MDM2 protein, the method comprising contacting a peptide with the MDM2 protein, wherein the peptide comprises at least two amino acids connected by a hydrocarbon covalent cross-link that stabilizes a secondary structure an α

-helix of the peptide.

2. The method of claim 1, further comprising contacting the peptide with the p53 protein.

3. The method of claim 1, wherein the peptide comprises an MDM2 binding site of a p53 donor helix.

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

5. The method of claim 1, wherein at least one amino acid in the peptide is a non-natural amino acid.

6. The method of claim 1, wherein the at least two amino acids in the peptide comprise olefins.

7. 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.

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

9. The method of claim 7, wherein at least one amino acid that is connected by the hydrocarbon covalent cross-link comprises an alpha-methyl, alpha-alkenyl amino acid.

10. The method of claim 9, wherein the alpha-methyl, alpha-alkenyl amino acid comprises a n-pentenyl or n-octenyl side chain.

11. The method of claim 1, wherein the cross-link spans from one to two turns on an α

-helix.

12. The method of claim 1, wherein the length of the cross-link between the two α

positions is 7 or 8 carbon atoms.

13. The method of claim 1, wherein the length of the cross-link between the two α

positions is 9 to 12 carbon atoms.

14. The method of claim 1, wherein the length of the cross-link between the two α

positions is 11 carbon atoms.

15. The method of claim 1, wherein the cross-linked peptide exhibits increased helicity compared to an uncrosslinked peptide.

16. The method of claim 1, wherein the cross-link is generated by a transition metal catalyzed cross-coupling reaction.