Arthritis & Rheumatism, Volume 62,
November 2010 Abstract Supplement
Abstracts of the American College of
Rheumatology/Association of Rheumatology Health Professionals
Annual Scientific Meeting
Atlanta, Georgia November 6-11, 2010.
HMPL-011, a Small Molecule Compound, Modulates Pro-Inflammatory Cytokine Production, in Part, Via Potentiating IL-10 Production.
Gu2, Weihua, He2, Weigang, Qiu2, Yan, Zhou2, Zhenhao, Dai2, Xiaomin, Shen2, Wuzhong, Yang2, Xiaoning
HMPL-011, a small molecular compound, was shown to be effective in reducing clinical arthritis score and joint swelling in collagen induced arthritis (CIA) in mouse (ACR 2010 Abstract). An inhibition on pro-inflammatory cytokines, such as IL-1b, MCP-1 and IL-6, and an increase in anti-inflammatory cytokine IL-10 was found in the arthritic joints. In this study, the mechanism of action of HMPL-011 was investigated.
Cytokine mRNA was analyzed by RT-PCR and protein by ELISA or CBA (Cytometric Bead Array). Phosphorylated MKK3/6 and p38 in RAW264.7 was analyzed by WB. For LPS induced inflammation in vivo, female Balb/C mouse was p.o. treated with HMPL-011 followed by LPS stimulation 30 minutes post HMPL-011 administration. Protein and mRNA levels were assessed 2hr and 8hr after LPS dosing. In IL-10 neutralization assay, IL-10 neutralizing antibody (JES5-2A5) was dosed i.v. and concurrently p.o. treated with HMPL-011 for 30min, followed by LPS treatment. Plasma cytokines were analyzed as indicated above.
HMPL-011 was screened at 10 mM against 48 inflammation related targets, include cytokine release, cytokine, chemokine and glucocorticoid receptor binding, caspases, MMPs, phosphatase and kinase activity, T, B cell proliferation, and NF-kB, NF-AT transcription activity. No significant activity of HMPL-011 was identified, except for the inhibition of IL-1b and IFNg release (IC50=11.0 mM and 9.7 mM, respectively). Furthermore, in a screen against 60 enzymes consisting of receptor tyrosine kinases and other inflammation- related kinases at 10 mM, no significant inhibition was observed. T, B cell activation, macrophage phagocytosis, NK cell cytotoxicity was also not affected by HMPL-011. However, IL-10 expression was found to be elevated by HMPL-011 in LPS stimulated mouse macrophage cells, RAW264.7, and this induction was shown to be inhibited by the p38 inhibitor, SB203580. Moreover, LPS induced MKK3/6 and p38a phosphorylation was shown to be enhanced by HMPL-011 in RAW264.7. In LPS treated mouse, HMPL-011 increased IL-10 at 2hr post LPS treatment, followed by a significant, potent inhibition on IL-6, MCP-1 and IFNg at 8hr post LPS treatment. In IL-10 neutralized mouse, HMPL-011 anti-inflammatory effect was found to be partially reversed, indicating the anti-inflammatory effect of HMPL-011 is only partially mediated by IL-10.
HMPL-011's anti-inflammatory effect was mediated at least in part through enhancement of IL-10. HMPL-011 may represent a novel class of small molecular drug in arthritis treatment.
To cite this abstract, please use the following information:
Gu, Weihua, He, Weigang, Qiu, Yan, Zhou, Zhenhao, Dai, Xiaomin, Shen, Wuzhong, et al; HMPL-011, a Small Molecule Compound, Modulates Pro-Inflammatory Cytokine Production, in Part, Via Potentiating IL-10 Production. [abstract]. Arthritis Rheum 2010;62 Suppl 10 :274