MK2 interacting proteins

• US 7,364,870 B2
• Filed: 08/01/2003
• Issued: 04/29/2008
• Est. Priority Date: 08/02/2002
• Status: Expired due to Fees

• Current Assignee: Wyeth (Pfizer Inc.)
• Original Assignee: Wyeth (Pfizer Inc.)
• Inventors: Yannoni, Yvonne M., Lin, Lih-Ling
• Primary Examiner(s): Monshipouri; Maryam
• Application Number:
  US10/523,014
• Publication Number:
  US 20060094101A1
• Time in Patent Office: 1,733 Days
• Field of Search: 435/15, 435/194
• US Class Current: 435/15
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• 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 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/4702 Regulators; Modulating activity
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C12Q 1/485 involving kinase
• CPC Class Codes: G01N 2333/91215 with a definite EC number (2.7.1.-)
• CPC Class Codes: G01N 2500/00 Screening for compounds of potential therapeutic value
• CPC Class Codes: G01N 33/564 for pre-existing immune complex or autoimmune disease , i.e. systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, rheumatoid factors or complement components C1-C9
• CPC Class Codes: G01N 33/6872 Intracellular protein regulatory factors and their receptors, e.g. including ion channels
• CPC Class Codes: G01N 33/6893 related to diseases not provided for elsewhere

Abstract

The present invention relates to uses of proteins that bind MK2 to modulate inflammation. More particularly, the invention relates to uses of proteins that bind MK2 for treating condition that are related to inflammation. The invention is useful for treating inflammatory conditions, particularly those in which a decrease in inflammation would be therapeutically beneficial.

Claims

1. A screening assay to identify compounds that inhibit or promote formation of a protein complex, comprising

(i) providing a two-hybrid assay system comprising:

a) a first fusion protein comprising an MK2 polypeptide, wherein said MK2 polypeptide comprises a proline-rich region, a kinase catalytic domain, a threonine residue that can be phosphorylated by MAP kinase, and a nuclear localization signal; and

b) a second fusion protein comprising a Shc polypeptide set forth as SEQ ID NO;

6, under conditions wherein the two proteins interact in the two hybrid assay system;

(ii) measuring a level of interaction between the fusion proteins in the presence and in the absence of a test compound; and

(iii) comparing the level of interaction of the fusion proteins,

wherein a difference in the amount of complex in the presence of the test compound, relative to the amount of complex in the absence of the test compound indicates that the test compound inhibits or promotes complex formation.

2. The assay of claim 1, wherein an increase in the amount of complex in the presence of the test compound indicates that the test compound promotes complex formation.

3. The assay of claim 1, wherein a decrease in the amount of complex in the presence of the test compound indicates that the test compound inhibits complex formation.

4. The assay of claim 1, wherein the MK2 polypeptide comprises amino acids 1-370.

5. The assay of claim 1, wherein the MK2 polypeptide comprises amino acids 41-338.

6. The assay of claim 1, wherein the MK2 polypeptide has an arginine at position 93 in the ATP-binding pocket.