Methods for identification and uses of anti-inflammatory receptors for eicosapentaenoic acid analogs

• US 7,341,840 B2
• Filed: 09/01/2005
• Issued: 03/11/2008
• Est. Priority Date: 03/05/2003
• Status: Active Grant

• Current Assignee: Brigham and Women's Hospital Incorporated
• Original Assignee: Brigham and Women's Hospital Incorporated
• Inventors: Serhan, Charles N., Arita, Makoto
• Primary Examiner(s): MERTZ, PREMA MARIA
• Application Number:
  US11/218,281
• Publication Number:
  US 20060024758A1
• Time in Patent Office: 922 Days
• Field of Search: None
• US Class Current: 435/7.2
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: G01N 2333/4703 Regulators; Modulating activity
• CPC Class Codes: G01N 2333/715 for cytokines; for lymphokines; for interferons
• CPC Class Codes: G01N 2333/726 G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH
• 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 33/5023 on expression patterns
• CPC Class Codes: G01N 33/5032 on intercellular interactions
• CPC Class Codes: G01N 33/5041 involving analysis of members of signalling pathways
• CPC Class Codes: G01N 33/5047 Cells of the immune system
• CPC Class Codes: G01N 33/505 involving T-cells
• CPC Class Codes: G01N 33/566 using specific carrier or receptor proteins as ligand binding reagents where possible specific carrier or receptor proteins are classified with their target compounds

Abstract

The present invention is directed to methods for the identification and uses of receptors that interact with anti-inflammatory compounds derived from eicosapentaenoic acid (EPA). The receptors are of the G-protein coupled receptor (GPCR) family, and are useful to screen candidate substances for anti-inflammatory activity, especially substances that are analogs of EPA. Such analogs are termed “resolvins”; and are typically di- and tri-hydroxy EPA analogs. One analog herein denoted Resolvin E1 was identified in humans and prepared by total synthesis. In nanomolar range Resolvin E1 reduces dermal inflammation, peritonitis, dendritic cells (DCs) migration and IL-12 production. Also described herein is a receptor denoted Reso ER1 that interacts with Resolvin E1 to attenuate cytokine induced activation of inflammatory pathways mediated by transcription factor (NF)-kB. Treatment of DCs with small-interfering RNA specific for ResoE1 eliminated the ligand'"'"'s ability to regulate IL-12. Assays of anti-inflammatory activity based on these discoveries are also described.

Claims

1. A method for screening a candidate substance for anti-inflammatory activity, comprising

contacting a cell that expresses a Reso E receptor with the candidate substance;

and

detecting a biological activity mediated by the Reso E receptor, wherein the Reso E receptor is the murine Reso E receptor according to SEQ. ID NO;

2, a rat Reso E receptor according to SEQ. ID NO;

3 or human Reso ER1 according to SEQ. ID NO;

1, wherein the biological activity is reduced TNF-α

cytokine induced activation of NF-kB transcription factor relative to a cell not contacted with the candidate substance.

2. The method of claim 1 wherein the cell is a leucocyte.

3. The method of claim 2 wherein the leucocyte is a monocyte, neutrophil, or a T-lymphocyte.

4. The method of claim 3 wherein the leukocyte is a monocyte that is an APC macrophage or dendritic cell.

5. The method of claim 1 wherein the Reso E receptor is human Reso ER1.

6. The method of claim 1 wherein the candidate substance comprises a resolvin molecule.

7. The method of claim 6 wherein the resolvin molecule is a poly-hydroxylated eicosapentaenoic acid (pH-EPA).

8. The method of claim 6 wherein the resolvin molecule is Reso-E1.