Integration of Affinity Selection–Mass Spectrometry and Functional Cell-Based Assays to Rapidly Triage Druggable Target Space within the NF-κB Pathway. (July 2016)

• Record Type: Journal Article
• Authors: Kutilek, Victoria D.; Andrews, Christine L.; Richards, Matthew P.; Xu, Zangwei; Sun, Tianxiao; Chen, Yiping; Hashke, Andrew; Smotrov, Nadya; Fernandez, Rafael; Nickbarg, Elliott B.; Chamberlin, Chad; Sauvagnat, Berengere; Curran, Patrick J.; Boinay, Ryan; Saradjian, Peter; Allen, Samantha J.; Byrne, Noel; Elsen, Nathaniel L.; Ford, Rachael E.; Hall, Dawn L.; Kornienko, Maria; Rickert, Keith W.; Sharma, Sujata; Shipman, Jennifer M.; Lumb, Kevin J.; Coleman, Kevin; Dandliker, Peter J.; Kariv, Ilona; Beutel, Bruce
• Is Part Of: Journal of biomolecular screening. Volume 21:Number 6(2016)
• Page Start: 608
• Page End: 619
• Publication Date: 2016-07
• Subjects: ALIS -- high-throughput screening -- affinity selection -- mass spectrometry -- phenotypic screening -- NF-κB pathway -- NF-κB1/p105 -- TRAF5; Drugs -- Analysis -- Periodicals; Drugs -- Testing -- Periodicals; Biomolecules -- Analysis -- Periodicals; 572.36
• DOI: 10.1177/1087057116637353
• Languages: English
• ISSNs: 1087-0571

Abstract

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.