A Profiling Platform for the Identification of Selective Metalloprotease Inhibitors

Abstract

Although proteases represent an estimated 5% to 10% of potential drug targets, inhibitors for metalloproteases (MPs) account for only a small proportion of all approved drugs, failures of which have typically been associated with lack of selectivity. In this study, the authors describe a novel and universal binding assay based on an actinonin derivative and show its binding activities for several MPs and its lack of activity toward all the non-MPs tested. This newly developed assay would allow for the rapid screening for inhibitors of a given MP and for the selectivity profiling of the resulting hits. The assay has successfully enabled for the first time simultaneous profiling of 8 well-known inhibitors against a panel of selected MPs. Previously published activities for these inhibitors were confirmed, and the authors have also discovered new molecular targets for some of them. The authors conclude that their profiling platform provides a generic assay solution for the identification of novel metalloprotease inhibitors as well as their selectivity profiling using a simple and homogeneous assay. (Journal of Biomolecular Screening 2008:285-294)

ADAM metalloproteases promote a developmental switch in responsiveness to the axonal repellant Sema3A

“…Thus, we repeated the Nrp1 staining analyses with rat proprioceptive and cutaneous axons that were cultured in the presence of the broad-spectrum MP inhibitor, TAPI-1 [46][47][48][49] (Fig. 2 and Supplementary Fig.…”

Upregulation of Mer Receptor Tyrosine Kinase Signaling Attenuated Lipopolysaccharide-Induced Lung Inflammation

“…Moreover, cleavage of Mer was enhanced by treatment of macrophages with LPS and phorbol 12-myristate 13-acetate and was specifically inhibited by the ADAM17 inhibitor TAPI-0 (Sather et al, 2007). TAPI-0 is a peptide-based compound in which the hydroxamic group (a strong chelating moiety) interacts with the catalytic zinc of the ADAM family of proteases and consequently inhibits their activities (Balakrishnan et al, 2006;Antczak et al, 2008). Consistently, we found that in vivo exposure of lungs to LPS enhanced the production of soluble Mer protein (sMer) but decreased membrane-bound Mer expression in alveolar macrophages in spite of increased Mer mRNA expression (Lee et al, 2012a).…”

Acetylenic inhibitors of ADAM10 and ADAM17: In silico analysis of potency and selectivity

“…Surprisingly, given their similar phylogenetic profile [8], TMI-1 remains moderately selective for TACE over ADAM10, as shown by the values for K i in Table 1. A close inspection of the geometry surrounding the bound ligand in both TACE and ADAM10 provides a possible clue as to why this is the case.…”

“…While sharing only a 39% sequence identity phylogenetic profiling clearly identifies ADAM10 and ADAM17 as comprising a distinct and separate subfamily from the other ADAM proteins [8]. Both proteinases function by cleaving their substrates at an extracellular site proximal to the cell membrane, thereby releasing the soluble fragment from the cell surface.…”