Abstract
We have already demonstrated that celecoxib, a selective cyclooxygenase (COX)-2 inhibitor, has a proapoptotic effect on synovial fibroblasts obtained from patients with rheumatoid arthritis (RA). Here we report on the development of two novel derivatives of celecoxib, N-(2-aminoethyl)-4-[5-(4-tolyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide (TT101) and 4-[5-(4-aminophenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide (TT201), including whether these compounds have a proapoptotic effect on synovial fibroblasts. Synovial fibroblasts were harvested from the synovial tissues of patients with RA or osteoarthritis (OA). Cell proliferation and cell viability were assessed by the incorporation of 5-bromo-2′-deoxyuridine and by the 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium monosodium salt assay, respectively. Apoptosis was detected by the identification of DNA fragmentation, and activation of caspase-3 was detected by the addition of a caspase-3 substrate to cell lysates. Production of prostaglandin E2 by RA synovial fibroblasts was analyzed by enzyme-linked immunosorbent assay. TT101 inhibited the proliferation of RA and OA synovial fibroblasts in a concentration-dependent manner. It caused a marked decrease of cell viability and induced DNA fragmentation more potently than either celecoxib or SC-236 (4-[5-(4-chlorophenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide). TT101 also increased caspase-3 activity. The order of potency of the COX-2 inhibitory activity of these drugs in RA synovial fibroblasts was celecoxib = SC-236 > rofecoxib > TT201 > TT101. In conclusion, we developed TT101 with about a 5- to 10-fold stronger proapoptotic effect on RA and OA synovial fibroblasts compared with that of celecoxib. Although the mechanism of action of TT101 remains unclear, it may have potential as a novel antirheumatic agent.
Footnotes
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This work was supported in part by research grants from The Japanese Ministry of Education, Culture, Sports, Science and Technology (no. 14572167 to S.K.) and a Research Promotion grant from Toho University Graduate School of Medicine (no. 04-01 to S.K.).
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Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.
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doi:10.1124/jpet.105.086116.
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ABBREVIATIONS: NSAIDs, nonsteroidal anti-inflammatory drugs; COX, cyclooxygenase; RA, rheumatoid arthritis; PG, prostaglandin; TT101, N-(2-aminoethyl)-4-[5-(4-tolyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide; TT201, 4-[5-(4-aminophenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide; DMSO, dimethyl sulfoxide; SC-236, 4-[5-(4-chlorophenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzene sulfonamide; IL-1β, interleukin-1β; FBS, fetal bovine serum; OA, osteoarthritis; BrdU, 5-bromo-2′-deoxyuridine; WST-1,4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzenedisulfonate; ELISA, enzyme-linked immunosorbent assay; PBS, phosphate-buffered saline; Pipes, 1,4-piperazinediethanesulfonic acid; Chaps, 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid; TBS, Tris-buffered saline; TSB-T, TBS containing 0.1% Tween 20; tBID, truncated BID; Z-LEHD-FMK, N-benzyloxycarbonyl-Leu-Glu-His-Asp-fluoromethyl ketone; Z-DEVD-FMK, N-benzyloxycarbonyl-Asp-Glu-Val-Asp-fluoromethyl ketone; Z-IETD-FMK, N-benzyloxycarbonyl-Ile-Glu-Thr-Asp-fluoromethyl ketone.
- Received March 9, 2005.
- Accepted April 29, 2005.
- The American Society for Pharmacology and Experimental Therapeutics
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