Intracellular Binding/Unbinding Kinetics of Approved Drugs to Carbonic Anhydrase II Observed by in-Cell NMR
| Autor: | Enrico Luchinat, Letizia Barbieri, Lucia Banci, Alessio Nocentini, Claudiu T. Supuran, Matteo Cremonini |
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| Přispěvatelé: | Luchinat, Enrico, Barbieri, Letizia, Cremonini, Matteo, Nocentini, Alessio, Supuran, Claudiu T, Banci, Lucia |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Drug Cell Membrane Permeability Proton Magnetic Resonance Spectroscopy Carbonic anhydrase II media_common.quotation_subject Drug Evaluation Preclinical Drug design Context (language use) Carbonic Anhydrase II 01 natural sciences Biochemistry Carbonic Anhydrase 03 medical and health sciences In vivo Carbonic anhydrase Drug Discovery Humans Carbonic Anhydrase Inhibitors Nuclear Magnetic Resonance Biomolecular media_common Dose-Response Relationship Drug biology in-cell NMR 010405 organic chemistry Chemistry Cell Membrane Articles General Medicine In vitro 0104 chemical sciences 3. Good health Kinetics HEK293 Cells 030104 developmental biology biology.protein Biophysics Molecular Medicine Intracellular Protein Binding |
| Zdroj: | ACS Chemical Biology |
| ISSN: | 1554-8937 1554-8929 |
| DOI: | 10.1021/acschembio.0c00590 |
| Popis: | Candidate drugs rationally designed in vitro often fail due to low efficacy in vivo caused by low tissue availability or because of unwanted side effects. To overcome the limitations of in vitro rational drug design, the binding of candidate drugs to their target needs to be evaluated in the cellular context. Here, we applied in-cell NMR to investigate the binding of a set of approved drugs to the isoform II of carbonic anhydrase (CA) in living human cells. Some compounds were originally developed toward other targets and were later found to inhibit CAs. We observed strikingly different dose- and time-dependent binding, wherein some drugs exhibited a more complex behavior than others. Specifically, some compounds were shown to gradually unbind from intracellular CA II, even in the presence of free compound in the external medium, therefore preventing the quantitative formation of a stable protein-ligand complex. Such observations could be correlated to the known off-target binding activity of these compounds, suggesting that this approach could provide information on the pharmacokinetic profiles of lead candidates at the early stages of multitarget drug design. |
| Databáze: | OpenAIRE |
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