Integration of Affinity Selection–Mass Spectrometry and Functional Cell-Based Assays to Rapidly Triage Druggable Target Space within the NF-κB Pathway
| Autor: | Nathaniel L. Elsen, Zangwei Xu, Peter J. Dandliker, Peter Saradjian, Keith W. Rickert, Berengere Sauvagnat, Nadya Smotrov, Dawn L. Hall, Elliott B. Nickbarg, Yiping Chen, Sujata Sharma, Kevin J. Lumb, Maria Kornienko, Noel Byrne, Chad Chamberlin, Christine Andrews, Samantha J Allen, Ilona Kariv, Jennifer M. Shipman, Patrick J. Curran, Kevin G. Coleman, Beutel Bruce A, Rachael E. Ford, Victoria Kutilek, Andrew Hashke, Rafael Fernandez, Tianxiao Sun, Matthew Richards, Ryan Boinay |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
High-throughput screening Phenotypic screening Cell Druggability Computational biology Biology Ligands Bioinformatics 01 natural sciences Biochemistry Mass Spectrometry Analytical Chemistry Structure-Activity Relationship 03 medical and health sciences Drug Discovery medicine TNF Receptor-Associated Factor 5 Reverse pharmacology 010405 organic chemistry Drug discovery NF-kappa B Transcription Factor RelA Ligand (biochemistry) Phenotype High-Throughput Screening Assays 0104 chemical sciences 030104 developmental biology medicine.anatomical_structure Molecular Medicine Protein Binding Signal Transduction Biotechnology |
| Zdroj: | SLAS Discovery. 21:608-619 |
| ISSN: | 2472-5552 |
| DOI: | 10.1177/1087057116637353 |
| Popis: | The primary objective of early drug discovery is to associate druggable target space with a desired phenotype. The inability to efficiently associate these often leads to failure early in the drug discovery process. In this proof-of-concept study, the most tractable starting points for drug discovery within the NF-κB pathway model system were identified by integrating affinity selection-mass spectrometry (AS-MS) with functional cellular assays. The AS-MS platform Automated Ligand Identification System (ALIS) was used to rapidly screen 15 NF-κB proteins in parallel against large-compound libraries. ALIS identified 382 target-selective compounds binding to 14 of the 15 proteins. Without any chemical optimization, 22 of the 382 target-selective compounds exhibited a cellular phenotype consistent with the respective target associated in ALIS. Further studies on structurally related compounds distinguished two chemical series that exhibited a preliminary structure-activity relationship and confirmed target-driven cellular activity to NF-κB1/p105 and TRAF5, respectively. These two series represent new drug discovery opportunities for chemical optimization. The results described herein demonstrate the power of combining ALIS with cell functional assays in a high-throughput, target-based approach to determine the most tractable drug discovery opportunities within a pathway. |
| Databáze: | OpenAIRE |
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