title: SEC-TID: A Label-Free Method for Small-Molecule Target Identification.
abstract: Bioactive small molecules are an invaluable source of therapeutics and chemical probes for exploring biological pathways. Yet, significant hurdles in drug discovery often come from lacking a comprehensive view of the target(s) for both early tool molecules and even late-stage drugs. To address this challenge, a method is provided that allows for assessing the interactions of small molecules with thousands of targets without any need to modify the small molecule of interest or attach any component to a surface. We describe size-exclusion chromatography for target identification (SEC-TID), a method for accurately and reproducibly detecting ligand-macromolecular interactions for small molecules targeting nucleic acid and several protein classes. We report the use of SEC-TID, with a library consisting of approximately 1000 purified proteins derived from the protein databank (PDB), to identify the efficacy targets tankyrase 1 and 2 for the Wnt inhibitor XAV939. In addition, we report novel interactions for the tumor-vascular disrupting agent vadimezan/ASA404 (interacting with farnesyl pyrophosphate synthase) and the diuretic mefruside (interacting with carbonic anhydrase XIII). We believe this method can dramatically enhance our understanding of the mechanism of action and potential liabilities for small molecules in drug discovery pipelines through comprehensive profiling of candidate druggable targets.
authors.affiliation: Developmental and Molecular Pathways Department, Novartis Institutes for Biomedical Research, Cambridge, MA, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Salcius
authors.affiliation: Developmental and Molecular Pathways Department, Novartis Institutes for Biomedical Research, Cambridge, MA, USA.
authors.forename: Andras J
authors.initials: AJ
authors.lastname: Bauer
authors.affiliation: Analytical Sciences, Novartis Institutes for Biomedical Research, Cambridge, MA, USA.
authors.forename: Qin
authors.initials: Q
authors.lastname: Hao
authors.affiliation: Analytical Sciences, Novartis Institutes for Biomedical Research, Cambridge, MA, USA.
authors.forename: Shu
authors.initials: S
authors.lastname: Li
authors.affiliation: Developmental and Molecular Pathways Department, Novartis Institutes for Biomedical Research, Cambridge, MA, USA.
authors.forename: Antonin
authors.initials: A
authors.lastname: Tutter
authors.affiliation: Developmental and Molecular Pathways Department, Novartis Institutes for Biomedical Research, Cambridge, MA, USA.
authors.forename: Jacob
authors.initials: J
authors.lastname: Raphael
authors.affiliation: Center for Proteomic Chemistry, Novartis Institutes for Biomedical Research, Basel, Switzerland.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Jahnke
authors.affiliation: Center for Proteomic Chemistry, Novartis Institutes for Biomedical Research, Basel, Switzerland.
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Rondeau
authors.affiliation: Center for Proteomic Chemistry, Novartis Institutes for Biomedical Research, Basel, Switzerland.
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Bourgier
authors.affiliation: Developmental and Molecular Pathways Department, Novartis Institutes for Biomedical Research, Cambridge, MA, USA.
authors.forename: John
authors.initials: J
authors.lastname: Tallarico
authors.affiliation: Developmental and Molecular Pathways Department, Novartis Institutes for Biomedical Research, Cambridge, MA, USA gregory.michaud@novartis.com.
authors.forename: Gregory A
authors.initials: GA
authors.lastname: Michaud
mesh_terms: D000818:Animals; D002151:Calorimetry; D020134:Catalytic Domain; D002417:Cattle; D002850:Chromatography, Gel / Q000379:methods*; D002853:Chromatography, Liquid; D018360:Crystallography, X-Ray; D030562:Databases, Protein; D055808:Drug Discovery; D005470:Fluorometry; D006801:Humans; D008024:Ligands; D046911:Macromolecular Substances; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry / Q000379:methods*; D009696:Nucleic Acids / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D040901:Proteomics / Q000379:methods*; D054852:Small Molecule Libraries / Q000737:chemistry; D044004:Xanthones / Q000737:chemistry
chemical_list: D008024:Ligands; D046911:Macromolecular Substances; D009696:Nucleic Acids; D011506:Proteins; D054852:Small Molecule Libraries; D044004:Xanthones; C066668:vadimezan
keywords: liquid chromatography–mass spectrometry; nucleic acids; proteins; size-exclusion chromatography; small molecules; target identification