(E)-2(R)-[1(S)-(Hydroxycarbamoyl)-4-phenyl-3-butenyl]-2′-isobutyl-2′-(methanesulfonyl)-4-methylvalerohydrazide (Ro 32-7315), a Selective and Orally Active Inhibitor of Tumor Necrosis Factor-α Convertase

Abstract

Tumor necrosis factor-␣ (TNF-␣), a cytokine secreted by inflammatory cells, has been implicated in several inflammatory disease, is a potent, orally active inhibitor of the TNF-␣ convertase (TACE), an enzyme responsible for proteolytic cleavage of the membrane bound precursor, pro-TNF-␣. Ro 32-7315 inhibited a recombinant form of TACE (IC 50 ϭ 5.2 nM) with selectivity over related matrix metalloproteinases. In a cellular assay system, THP-1 cell line, and in human and rat whole blood, Ro 32-7315 significantly reduced lipopolysaccharide (LPS)-induced TNF-␣ release with IC 50 values of 350 Ϯ 14 nM (n ϭ 5), 2.4 Ϯ 0.5 M (n ϭ 5), and 110 Ϯ 18 nM (n ϭ 5), respectively. Oral administration of Ro 32-7315 to Wistar rats caused a dose-dependent inhibition of LPS-induced release of systemic TNF-␣ with an ED 50 of 25 mg/kg. Treatment (days 0 -14) of Allen and Hamburys hooded rats with Ro 32-7315 (2.5, 5, 10, and 20 mg/kg, i.p., twice daily) significantly reduced adjuvant-induced secondary paw swelling (42, 71, 83, and 93%, respectively) as compared with the vehicle group. In the Ro 32-7315-treated group, the reduced paw swelling was associated with improved lesion score and joint mobility. Furthermore, in a placebocontrolled, single-dose study, Ro 32-7315 given orally (450 mg) significantly suppressed ex vivo, LPS-induced TNF-␣ release in the whole-blood samples taken from healthy male and female volunteers (mean inhibition of 42% over a 4-h duration, n ϭ 6). These data collectively support the potential use of such a compound for the oral treatment of inflammatory disorders.

Modulation of autocrine TNF-α-stimulated matrix metalloproteinase 9 (MMP-9) expression by mitogen-activated protein kinases in THP-1 monocytic cells

“…The importance of released TNF-a in this context was also shown by others using the unspecific metalloproteinase inhibitor Marimastat (Robinson et al, 2002). Applying the specific TACE inhibitor Ro32-7315 (Beck et al, 2002), we demonstrated that TACE activity is essential for autocrine TNF-a-stimulated MMP-9 expression. Slight impairment of MMP-9 gene expression was also observed upon blocking of TNF-R2, which may be caused by 'ligand passing' from TNF-R2 to TNF-R1.…”

“…To investigate whether TACE, which is responsible for the release of TNF-a from the plasma membrane, is involved in basal MMP-9 secretion, THP-1 cells were treated with Ro32-7315, a highly specific inhibitor of TACE activity (Beck et al, 2002). Zymographic analysis of secreted enzymes showed that increasing concentrations of Ro32-7315 led to a decrease in MMP-9 release from the cells ( Figure 1E).…”

TNF-α-converting enzyme (TACE/ADAM17)-dependent loss of CD30 induced by proteasome inhibition through reactive oxygen species

“…Although 100 ng/ml of bortezomib doubled the sCD30 release (707.5-1569 U/ml, Po0.0001) in ADAM10-defective cells with functional ADAM17, it failed to stimulate CD30 shedding in ADAM17-defective cells. We also applied the ADAM17-selective inhibitor, Ro32-7315 13 ( Figure 2b). It blocked bortezomib-stimulated sCD30 release from L540 cells with an IC 50 of approximately 230 nM.…”

“…Dot blot analysis shows the percentage of viable (lower lift) and apoptotic (upper and lower right) cells. 13.31% (upper right gate). These data confirm that induction of CD30 shedding by bortezomib substantially perturbs anti-CD30-targeted immunotherapy.…”

“…GI254023X (GlaxoSmithKline, Stevenage, UK) was kindly provided by Dr A Ludwig, Aachen, Germany. The ADAM17-selective inhibitor, Ro32-7315, 13 and the gelatinase-selective inhibitor, Ro28-2653, were from Roche Diagnostics GmbH, Pharma Research (Penzberg, Germany).…”