Abstract
Demonstration that IκB kinase 2 (IKK-2) plays a pivotal role in the nuclear factor-κB-regulated production of proinflammatory molecules by stimuli such as tumor necrosis factor (TNF)-α and interleukin (IL)-1 suggests that inhibition of IKK-2 may be beneficial in the treatment of rheumatoid arthritis. In the present study, we demonstrate that a novel, potent (IC50 = 17.9 nM), and selective inhibitor of human IKK-2, 2-[(aminocarbonyl)amino]-5-(4-fluorophenyl)-3-thiophenecarboxamide (TPCA-1), inhibits lipopolysaccharide-induced human monocyte production of TNF-α, IL-6, and IL-8 with an IC50 = 170 to 320 nM. Prophylactic administration of TPCA-1 at 3, 10, or 20 mg/kg, i.p., b.i.d., resulted in a dose-dependent reduction in the severity of murine collagen-induced arthritis (CIA). The significantly reduced disease severity and delay of disease onset resulting from administration of TPCA-1 at 10 mg/kg, i.p., b.i.d. were comparable to the effects of the antirheumatic drug, etanercept, when administered prophylactically at 4 mg/kg, i.p., every other day. Nuclear localization of p65, as well as levels of IL-1β, IL-6, TNF-α, and interferon-γ, were significantly reduced in the paw tissue of TPCA-1- and etanercept-treated mice. In addition, administration of TPCA-1 in vivo resulted in significantly decreased collagen-induced T cell proliferation ex vivo. Therapeutic administration of TPCA-1 at 20 mg/kg, but not at 3 or 10 mg/kg, i.p., b.i.d., significantly reduced the severity of CIA, as did etanercept administration at 12.5 mg/kg, i.p., every other day. These results suggest that reduction of proinflammatory mediators and inhibition of antigen-induced T cell proliferation are mechanisms underlying the attenuation of CIA by the IKK-2 inhibitor, TPCA-1.
Footnotes
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doi:10.1124/jpet.104.074484.
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ABBREVIATIONS: RA, rheumatoid arthritis; IKK, IκB kinase; NF-κB, nuclear factor-κB; TNF, tumor necrosis factor; IL, interleukin; TPCA-1, 2-[(aminocarbonyl)amino]-5-(4-fluorophenyl)-3-thiophenecarboxamide; LPS, lipopolysaccharide; CIA, collagen-induced arthritis; IFN, interferon; COX, cyclooxygenase; MMP, matrix metalloproteinase; CHAPS, 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid; DMSO, dimethyl sulfoxide; DMA, dimethylacetoacetamide; LNC, lymph node cells; ELISA, enzyme-linked immunosorbent assay; JNK3, c-Jun N-terminal kinase 3; AUC, area under the curve; NSAIDs, nonsteroidal anti-inflammatory drugs; DMARDs, disease-modifying antirheumatic drugs; GST, glutathione S-transferase; BSA, bovine serum albumin; DTT, dithiothreitol; HBSS, Hanks' balanced salt solution; APC, antigen-presenting cells; Th1, subset 1 of T helper cells.
- Received July 20, 2004.
- Accepted August 9, 2004.
- The American Society for Pharmacology and Experimental Therapeutics