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A Novel small molecule Inhibitor to C‐Reactive Protein attenuates CRP's Pro‐Inflammatory Effects In‐Vivo
Author(s) -
Jialal Ishwarlal,
Kumaresan Pappanaicken R,
Lam Kit S,
Devaraj Sridevi
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.379.12
Subject(s) - in vivo , c reactive protein , chemistry , pharmacology , in vitro , superoxide , human serum albumin , reactive oxygen species , inflammation , biochemistry , immunology , medicine , biology , enzyme , microbiology and biotechnology
Elevated C‐reactive protein (CRP) levels predict cardiovascular disease and augur a poor prognosis in patients with acute coronary syndromes. Much in vitro and in vivo data support a role for CRP in atherogenesis. There is an urgent need to develop inhibitors that specifically block the biological effects of CRP in vivo. Using the one‐bead‐one‐compound (OBOC) combinatorial library method, we recently used a novel fluorescent based screening method to screen OBOC combinatorial library for the discovery of peptides against human CRP and wished to examine effects in vitro and in vivo. Human CRP was labeled with FITC and Human serum albumin (HuSA) was labeled with PE and used for screening. The OBOC library LWH‐01 was synthesized on TentaGel resin beads using standard solid phase “split/mix” approach. By subtraction screening, 8 peptides that bind specifically to CRP and not to human serum albumin (HuSA) were identified. In HAEC incubated with CRP, inhibitors CRPi‐2, CRPi‐3 and CRPi‐6 significantly inhibited CRP‐induced superoxide, cytokine release and nuclear NFκb activity (p<0.05). Molecular docking studies demonstrate that CRPi‐2 interacts with the two Ca2+ ions in the single subunit of CRP. Treatment of CRP‐injected Wistar rats for 4 days with CRPi‐2 resulted in significant reduction in superoxide release, cytokines (IL‐1, TNF, IL‐8 and MCP‐1) release and NFκb activity compared to rats treated with CRP alone (p<0.05). There were no toxic effects as evidenced by routine clinical laboratory tests. In conclusion, we identify a novel inhibitor to CRP which attenuates its biological effects both in‐vitro and in‐vivo and in future studies will examine the utility of this inhibitor in longer term studies in animal models.