Method of determining inhibition of binding to TRPM7 protein

• US 7,858,322 B2
• Filed: 12/22/2004
• Issued: 12/28/2010
• Est. Priority Date: 12/23/2003
• Status: Active Grant

• Current Assignee: NoNO, Inc.
• Original Assignee: Arbor Vita Corporation, NoNO, Inc.
• Inventors: Garman, Jonathan David, Belmares, Michael P., Tymianski, Michael
• Primary Examiner(s): Chernyshev; Olga N
• Application Number:
  US11/021,928
• Publication Number:
  US 20050164933A1
• Time in Patent Office: 2,197 Days
• Field of Search: None
• 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: C07K 14/47 from mammals
• CPC Class Codes: C07K 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C07K 2319/10 containing a tag for extracellular membrane crossing, e.g. TAT or VP22
• CPC Class Codes: C12N 15/1138 against receptors or cell surface proteins
• CPC Class Codes: C12N 2799/022 where the vector is derived from an adenovirus
• CPC Class Codes: G01N 2500/00 Screening for compounds of potential therapeutic value

Abstract

This invention relates to methods of reducing the damaging effect of an injury to mammalian cells by treatment with compounds which reduce cell death or dysfunction, including cellular damage following episodes of tissue ischemia, trauma, epilepsy, and acute or chronic degeneration. The invention discloses methods of treating these disorders by administering inhibitors that disrupt protein-protein interactions involved in these disorders, screening methods to identify such inhibitors and specific compositions useful for treating these disorders.

Claims

1. A method for determining whether a test compound inhibits binding between a TRPM7 protein and a PDZ domain-containing polypeptide, comprising:

(i) contacting a TRPM7 protein or a fragment thereof containing at least the four C-terminal amino acids of the protein with a PDZ domain-containing polypeptide in the presence and absence of the test compound; and

(ii) measuring the amount of complex formed between the TRPM7 protein or fragment thereof and the PDZ domain-containing polypeptide in the presence and absence of the test compound,

wherein less complex is formed in the presence relative to the absence of the test compound indicating the test compound is an inhibitor of binding between the TRPM7 protein and the PDZ domain-containing polypeptide.

2. The method of claim 1, wherein the PDZ domain is a PDZ domain selected from the group consisting of RIM-2, Mint1, INADL, Syntrophin 1 alpha,SITAC-18, LIM mystique, ZO-1, PAR3L, MAST2, PAR3, and novel serine protease (SEQ ID NO:

158).

3. The method of claim 1, wherein the test compound is a peptide comprising at least two contiguous amino acids from the C-terminus of SEQ ID NO:

260 or SEQ ID NO;

384.

4. The method of claim 1, wherein the test compound comprises 4 residues from the C-terminus of SEQ ID NO:

260 or SEQ ID NO;

384.

5. The method of claim 1, wherein the test compound further comprises a transmembrane transporter peptide.

6. The method of claim 1, wherein the PDZ domain is a syntrophin-1 alpha PDZ domain.