Discovery of an enzyme and substrate selective inhibitor of ADAM10 using an exosite-binding glycosylated substrate
| Autor: | Radleigh G. Santos, Gregg B. Fields, Louis Scampavia, Jean-Phillipe Pettiloud, Andreas Ludwig, Daniela Dreymüller, Dmitriy Minond, Mare Cudic, Christoph Becker-Pauly, Franck Madoux, Thomas D. Bannister, Timothy P. Spicer |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Glycosylation Science ADAM17 Protein Article Substrate Specificity ADAM10 Protein Structure-Activity Relationship 03 medical and health sciences chemistry.chemical_compound In vivo Cell Line Tumor Humans Structure–activity relationship Binding site chemistry.chemical_classification Multidisciplinary biology Active site Substrate (chemistry) High-Throughput Screening Assays 3. Good health 030104 developmental biology Enzyme chemistry Biochemistry Drug development biology.protein Medicine Databases Chemical |
| Zdroj: | Scientific Reports Scientific Reports, Vol 6, Iss 1, Pp 1-17 (2016) Scientific reports 6, 11 (2016). doi:10.1038/s41598-016-0013-4 |
| ISSN: | 2045-2322 |
| DOI: | 10.1038/s41598-016-0013-4 |
| Popis: | ADAM10 and ADAM17 have been shown to contribute to the acquired drug resistance of HER2-positive breast cancer in response to trastuzumab. The majority of ADAM10 and ADAM17 inhibitor development has been focused on the discovery of compounds that bind the active site zinc, however, in recent years, there has been a shift from active site to secondary substrate binding site (exosite) inhibitor discovery in order to identify non-zinc-binding molecules. In the present work a glycosylated, exosite-binding substrate of ADAM10 and ADAM17 was utilized to screen 370,276 compounds from the MLPCN collection. As a result of this uHTS effort, a selective, time-dependent, non-zinc-binding inhibitor of ADAM10 with Ki = 883 nM was discovered. This compound exhibited low cell toxicity and was able to selectively inhibit shedding of known ADAM10 substrates in several cell-based models. We hypothesize that differential glycosylation of these cognate substrates is the source of selectivity of our novel inhibitor. The data indicate that this novel inhibitor can be used as an in vitro and, potentially, in vivo, probe of ADAM10 activity. Additionally, results of the present and prior studies strongly suggest that glycosylated substrate are applicable as screening agents for discovery of selective ADAM probes and therapeutics. |
| Databáze: | OpenAIRE |
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