Stabilized alpha helical peptides and uses thereof

• US 8,198,405 B2
• Filed: 07/30/2008
• Issued: 06/12/2012
• Est. Priority Date: 11/05/2003
• Status: Active Grant

• Current Assignee: Dana-Farber Cancer Institute, President and Fellows of Harvard College (Harvard Corporation)
• Original Assignee: Dana-Farber Cancer Institute, President and Fellows of Harvard College (Harvard Corporation)
• Inventors: Walensky, Loren D., Korsmeyer, Stanley J., Verdine, Gregory
• Primary Examiner(s): Lukton, David
• Application Number:
  US12/182,673
• Publication Number:
  US 20090176964A1
• Time in Patent Office: 1,413 Days
• Field of Search: 514/2, 530/324, 530/317, 530/323
• US Class Current: 530/324
• 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: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/14 for treating abnormal movements, e.g. chorea, dyskinesia
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 35/04 specific for metastasis
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 5/06 of the anterior pituitary hormones, e.g. TSH, ACTH, FSH, LH, PRL, GH
• CPC Class Codes: A61P 5/14 : of the thyroid hormones, e.g. T3, T4
• CPC Class Codes: A61P 5/24 : of the sex hormones
• CPC Class Codes: A61P 7/00 : Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 7/02 : Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: A61P 7/06 : Antianaemics
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/04 : Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• CPC Class Codes: A61P 9/08 : Vasodilators for multiple indications
• CPC Class Codes: A61P 9/10 : for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: A61P 9/12 : Antihypertensives
• CPC Class Codes: A61P 9/14 : Vasoprotectives; Antihaemorrhoidals; Drugs for varicose therapy; Capillary stabilisers
• CPC Class Codes: C07K 1/113 : without change of the primary structure
• CPC Class Codes: C07K 1/1136 : by reversible modification of the secondary, tertiary or quarternary structure, e.g. using denaturating or stabilising agents
• CPC Class Codes: C07K 14/001 : by chemical synthesis
• CPC Class Codes: C07K 14/435 : from animals; from humans
• CPC Class Codes: C07K 14/4747 : Apoptosis related proteins
• CPC Class Codes: C07K 7/08 : having 12 to 20 amino acids gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68
• CPC Class Codes: C07K 7/64 : Cyclic peptides containing only normal peptide links

Abstract

Novel polypeptides and methods of making and using the same are described herein. The polypeptides include cross-linking (“hydrocarbon stapling”) moieties to provide a tether between two amino acid moieties, which constrains the secondary structure of the polypeptide. The polypeptides described herein can be used to treat diseases characterized by excessive or inadequate cellular death.

Claims

1. A cross-linked polypeptide comprising an amino acid sequence which is at least 60% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:

92-111, wherein the polypeptide comprises a crosslinker connecting a first amino acid to a second amino acid, and wherein the first and second amino acids are separated by 3 or 6 amino acids.

10. The cross-linked polypeptide of claim 1, wherein the polypeptide has the Formula:

11. The cross-linked polypeptide of claim 10, wherein the polypeptide has a substantially alpha helical secondary structure in aqueous solution.

12. The cross-linked polypeptide of claim 1, comprising an amino acid sequence which is at least 70% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:

92-111.

13. The cross-linked polypeptide of claim 1, comprising an amino acid sequence which is at least 80% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:

92-111.

14. The cross-linked polypeptide of claim 1, comprising an amino acid sequence which is at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:

92-111.

15. The cross-linked polypeptide of claim 1, wherein the cross-linked polypeptide comprises up to 50 amino acids.

19. The cross-linked polypeptide of claim 1, 2, 3, 4, or 5, wherein the polypeptide has a substantially alpha-helical secondary structure in aqueous solution.

2. A cross-linked polypeptide comprising an amino acid sequence which is at least 80% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:

92-111 and wherein the cross-linked polypeptide comprises up to 50 amino acids.

3. The cross-linked polypeptide of claim 2, wherein the cross-linked polypeptide comprises an amino acid sequence that is at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:

92-111 and wherein the cross-linked polypeptide comprises up to 50 amino acids.

4. The cross-linked polypeptide of claim 2, wherein the cross-linked polypeptide comprises an amino acid sequence that is at least 80% identical to SEQ ID NO:

111 and wherein the cross-linked polypeptide comprises up to 50 amino acids.

5. The cross-linked polypeptide of claim 3, wherein the cross-linked polypeptide comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:

111 and wherein the cross-linked polypeptide comprises up to 50 amino acids.

6. The cross-linked polypeptide of claim 5, wherein the cross-linked polypeptide comprises SEQ ID NO:

111.

7. The cross-linked polypeptide of claim 6, wherein the amino acid sequence of the cross-linked polypeptide is IWIAQELR*IGD*FNAYYARR (SEQ ID NO:

111) and wherein * is a tethered amino acid.

8. The cross-linked polypeptide of claim 7, wherein an alpha carbon of the tethered amino acid is in an S configuration.

9. The cross-linked polypeptide of claim 8, wherein a tether connecting the alpha carbons of the tethered amino acids contains nine consecutive C—

C bonds.

16. The cross-linked polypeptide of claim 2, 3, 4, or 5, wherein the cross-linked polypeptide comprises a crosslinker connecting a first amino acid to a second amino acid.

17. The cross-linked polypeptide of claim 16, wherein the first and second amino acids are separated by 3 or 6 amino acids.

18. The cross-linked polypeptide of claim 16, wherein the crosslinker is a hydrocarbon crosslinker.