Stabilized compounds having secondary structure motifs

• US 7,192,713 B1
• Filed: 05/18/2000
• Issued: 03/20/2007
• Est. Priority Date: 05/18/1999
• Status: Expired due to Term

• Current Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Original Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Inventors: Schafmeister, Christian E., Verdine, Gregory L.
• Primary Examiner(s): Tate; Christopher R.
• Assistant Examiner(s): CHISM, BILLY D
• Application Number:
  US09/574,086
• Time in Patent Office: 2,497 Days
• Field of Search: 435/7.1, 514/14, 530/300, 530/326, 530/327, 530/331, 530/332, 530/333, 530/334, 530/335, 530/336, 530/337, 530/338, 556/136
• US Class Current: 435/7.1
• 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 said peptide comprises at least two moieties capable of undergoing reaction to promote carbon-carbon bond formation; and (2) contacting said 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 said crosslinked stabilized α-helical structure with MDM2.

Claims

1. A method for synthesizing stabilized alpha helix structures comprising:

synthesizing a peptide from a selected number of standard or non-standard amino acids, wherein said peptide comprises at least two moieties capable of undergoing reaction to promote carbon-carbon bond formation; and

contacting said peptide with a reagent to generate at least one cross-linker between said at least two moieties and to effect stabilization of a alpha helix structure, wherein the length and geometry of said cross-linker is optimized to improve the yield of helix content and stability of said alpha helix structure.

2. The method of claim 1, wherein synthesizing a peptide comprises synthesizing the peptide fragment that forms the donor helix in the native complex p53 having at least two vinyl amino acids incorporated therein.

3. The method of claim 1, wherein synthesizing a peptide comprises synthesizing the peptide fragment that forms the donor helix in the native complex p53 having at least one divinyl amino acid and at least two vinyl amino acids incorporated therein.

4. The method of claim 3, wherein said cross-linker comprises 11 carbon atoms.

5. The method of claim 1, further comprising hydrogenating the peptide after said contacting step.

6. The method of claim 5, further comprising purifying the peptide.

7. The method of claim 1, further comprising purifying the peptide.

8. The method of claim 1, wherein said method is performed in solution.

9. The method of claim 1, wherein said method is performed on a solid support.

10. The method of claim 1, wherein said moieties in said peptide are unnatural amino acids.

11. The method of claim 1, wherein said peptide comprises two moieties, wherein the first moiety is at a desired position i along the peptide and the second moiety is at position i+7 from the first moiety.

12. The method of claim 11, wherein said cross-linker comprises at least 8 carbon atoms.

13. The method of claim 12, wherein said cross-linker comprises at least 10 carbon atoms.

14. The method of claim 13, wherein said cross-linker comprises 11 carbon atoms.

15. The method of claim 1, wherein synthesizing the peptide comprises synthesizing a portion of the peptide fragment that forms the donor helix in the native complex p53 having at least two vinyl amino acids incorporated therein.

16. The method of claim 1, wherein synthesizing the peptide comprises synthesizing a portion of the peptide fragment that forms the donor helix in the native complex p53 having at least one divinyl amino acid and at least two vinyl amino acids incorporated therein.

17. The method of claim 1, wherein synthesizing the peptide comprises synthesizing a peptide that comprises the MDM2 binding site of the donor helix in the native complex p53.

18. The method of claim 17, wherein synthesizing the peptide comprises synthesizing the peptide that comprises the MDM2 binding site of the donor helix in the native complex p53 having at least two vinyl amino acids incorporated therein.

19. The method of claim 17, wherein synthesizing the peptide comprises synthesizing the peptide that comprises the MDM2 binding site of the donor helix in the native complex p53 having at least one divinyl amino acid and at least two vinyl amino acids incorporated therein.