Inhibitor Focusing: Direct Selection of Drug Targets from Proteomes Using Activity-Based Probes
| Autor: | Jonathan S. Rosenblum, Jonathan J. Burbaum, Arwin Aban, Tyzoon K. Nomanbhoy |
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| Rok vydání: | 2003 |
| Předmět: |
Drug
Proteome media_common.quotation_subject Dipeptidyl Peptidase 4 Organophosphonates Drug design Computational biology Biology Pharmacology In Vitro Techniques Kidney Binding Competitive Cell Line Mice Structure-Activity Relationship Drug Discovery Animals Humans Protease Inhibitors Binding site Isoleucine Selection (genetic algorithm) Cells Cultured media_common Fluorescent Dyes Binding Sites Drug discovery Drug candidate Membrane Proteins Proteins Small molecule Thiazoles Pharmaceutical Preparations Drug Design Molecular Medicine |
| Zdroj: | Assay and Drug Development Technologies. 1:137-146 |
| ISSN: | 1540-658X |
| Popis: | In the latter stages of drug discovery and development, assays that establish drug selectivity and toxicity are important when side effects, which are often due to lack of specificity, determine drug candidate viability. There has been no comprehensive or systematic methodology to measure these factors outside of whole-animal assays, and such phenomenological assays generally fail to establish the additional targets of a given small molecule, or the molecular origin of toxicity. Consequently, small-molecule development programs destined for failure often reach advanced stages of testing, and the money and time invested in such programs could be saved if information on selectivity were available early in the process. Here, we present a methodology that utilizes chemical ABPs in combination with small-molecule inhibitors to selectively label small-molecule binding sites in whole proteomic samples. In principle, the ABP and small molecule will compete for similar binding sites, such that the small molecule will protect against modification by the ABP. Thus, after removal of the small molecule, the binding site for the ABP will be revealed, and a second probe can then be used to label the small-molecule binding sites selectively. To demonstrate this experimentally, we mapped the binding sites of the DPP4 inhibitor, IT, in a number of different tissue types. |
| Databáze: | OpenAIRE |
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