BH4 stabilized peptides and uses thereof

• US 10,106,590 B2
• Filed: 03/14/2014
• Issued: 10/23/2018
• Est. Priority Date: 03/15/2013
• Status: Active Grant

• Current Assignee: Dana-Farber Cancer Institute
• Original Assignee: Dana-Farber Cancer Institute
• Inventors: Walensky, Loren D., Stewart, Michelle L., Barclay, Lauren
• Primary Examiner(s): Alstrum-Acevedo, James H
• Assistant Examiner(s): Dabkowski, Erinne R
• Application Number:
  US14/777,391
• Publication Number:
  US 20160031959A1
• Time in Patent Office: 1,684 Days
• 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 38/1761 Apoptosis related proteins, e.g. Apoptotic protease-activating factor-1 (APAF-1), Bax, Bax-inhibitory protein(s)(BI; bax-I), Myeloid cell leukemia associated protein (MCL-1), Inhibitor of apoptosis [IAP] or Bcl-2
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61K 49/0008 Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61K 9/08 Solutions composition of solutions A61K47/00
• CPC Class Codes: C07K 14/4747 Apoptosis related proteins
• CPC Class Codes: G01N 2333/47 Assays involving proteins of known structure or function as defined in the subgroups
• CPC Class Codes: G01N 2500/04 Screening involving studying the effect of compounds C directly on molecule A (e.g. C are potential ligands for a receptor A, or potential substrates for an enzyme A)
• CPC Class Codes: G01N 2500/20 cell-free systems
• CPC Class Codes: G01N 33/68 involving proteins, peptides or amino acids involving lipoproteins G01N33/92

Abstract

Provided herein are polypeptides containing stabilized BH4 domains of BCL-2 family proteins that are capable of binding and/or inactivating and/or modulating BAX protein, and/or its close homologs BAK and BOK, and/or other physiological BH4 targets. Also provided are compositions containing these polypeptides and methods of treating cytotoxic diseases that include administering to a subject one of the polypeptides.

Claims

1. An internally cross-linked polypeptide comprising the amino acid sequence of EIVMKYIHYKLSQRGYEWDA (SEQ ID NO:

1),

wherein;

the side chains of amino acid residues at positions 8 and 12 of SEQ ID NO;

1 or the side chains of amino acid residues at positions 13 and 17 of SEQ ID NO;

1 are replaced by an internal staple; and

the internally cross-linked polypeptide is capable of specifically binding to a BAX protein; and

optionally, the amino acid residue at position 4 of SEQ ID NO;

1 is substituted with another amino acid.

2. The cross-linked polypeptide of claim 1, wherein the side chains of amino acid residues 13 and 17 of SEQ ID NO:

1 are replaced by an internal staple.

3. A pharmaceutical composition comprising one or more of the cross-linked polypeptides of claim 1.

4. A kit for identifying agents that interact with and/or modulate the activity of BAX protein comprising one or more of the cross-linked polypeptides of claim 1 and a polypeptide comprising the amino acid sequence of a BAX polypeptide.

5. A compound comprising one or more of the cross-linked polypeptides of claim 1.

6. The cross-linked polypeptide of claim 1, wherein the side chains of the amino acid residues at positions 8 and 12 of SEQ ID NO:

1 are replaced by an internal staple.