title: Identification of small molecules that disrupt signaling between ABL and its positive regulator RIN1.
abstract: Constitutively active BCR-ABL kinase fusions are causative mutations in the pathogenesis of hematopoietic neoplasias including chronic myelogenous leukemia (CML). Although these fusions have been successfully targeted with kinase inhibitors, drug-resistance and relapse continue to limit long-term survival, highlighting the need for continued innovative drug discovery. We developed a time-resolved Förster resonance energy transfer (TR-FRET) -based assay to identify compounds that disrupt stimulation of the ABL kinase by blocking its ability to bind the positive regulator RIN1. This assay was used in a high throughput screen (HTS) of two small molecule libraries totaling 444,743 compounds. 708 confirmed hits were counter-screened to eliminate off-target inhibitors and reanalyzed to prioritize compounds with IC50 values below 10 μM. The CML cell line K562 was then used to identify five compounds that decrease MAPK1/3 phosphorylation, which we determined to be an indicator of RIN1-dependent ABL signaling. One of these compounds is a thiadiazole, and the other four are structurally related acyl piperidine amides. Notably, these five compounds lower cellular BCR-ABL1 kinase activity by blocking a positive regulatory interaction rather than directly inhibiting ABL catalytic function.
authors: Molecular Biology Institute, Jonsson Comprehensive Cancer Center, Department of Biological Chemistry, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, California, United States of America.
authors: Pamela Y
authors: PY
authors: Ting
authors: California NanoSystems Institute, University of California Los Angeles, Los Angeles, California, United States of America.
authors: Robert
authors: R
authors: Damoiseaux
authors: Department of Molecular and Medical Pharmacology, Crump Institute for Molecular Imaging, University of California at Los Angeles Metabolomics and Proteomics Center, California NanoSystems Institute and Jonsson Comprehensive Cancer Center, University of California Los Angeles, Los Angeles, California, United States of America.
authors: Björn
authors: B
authors: Titz
authors: Department of Microbiology, Immunology and Molecular Genetics, Jonsson Comprehensive Cancer Center, and California NanoSystems Institute, University of California Los Angeles, Los Angeles, California, United States of America.
authors: Kenneth A
authors: KA
authors: Bradley
authors: Department of Molecular and Medical Pharmacology, Crump Institute for Molecular Imaging, University of California at Los Angeles Metabolomics and Proteomics Center, California NanoSystems Institute and Jonsson Comprehensive Cancer Center, University of California Los Angeles, Los Angeles, California, United States of America.
authors: Thomas G
authors: TG
authors: Graeber
authors: The Scripps Research Institute-FL, Lead Identification, Translational Research Institute, Jupiter, Florida, United States of America.
authors: Virneliz
authors: V
authors: Fernández-Vega
authors: The Scripps Research Institute-FL, Department of Chemistry, Translational Research Institute, Jupiter, Florida, United States of America.
authors: Thomas D
authors: TD
authors: Bannister
authors: The Scripps Research Institute-FL, Lead Identification, Translational Research Institute, Jupiter, Florida, United States of America.
authors: Peter
authors: P
authors: Chase
authors: The Scripps Research Institute-FL, Department of Chemistry, Translational Research Institute, Jupiter, Florida, United States of America.
authors: Reji
authors: R
authors: Nair
authors: The Scripps Research Institute-FL, Lead Identification, Translational Research Institute, Jupiter, Florida, United States of America.
authors: Louis
authors: L
authors: Scampavia
authors: The Scripps Research Institute-FL, Lead Identification, Translational Research Institute, Jupiter, Florida, United States of America.
authors: Peter
authors: P
authors: Hodder
authors: The Scripps Research Institute-FL, Lead Identification, Translational Research Institute, Jupiter, Florida, United States of America.
authors: Timothy P
authors: TP
authors: Spicer
authors: Molecular Biology Institute, Jonsson Comprehensive Cancer Center, Department of Biological Chemistry, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, California, United States of America.
authors: John
authors: J
authors: Colicelli
mesh_terms: D044946:Biflavonoids / Q000494:pharmacology; D002392:Catechin / Q000031:analogs & derivatives / Q000494:pharmacology; D031541:Fluorescence Resonance Energy Transfer; D016044:Fusion Proteins, bcr-abl / Q000378:metabolism; D057166:High-Throughput Screening Assays; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism*; D020014:K562 Cells; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D011485:Protein Binding / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D016315:Proto-Oncogene Proteins c-abl / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015203:Reproducibility of Results; D015398:Signal Transduction / Q000187:drug effects*; D054852:Small Molecule Libraries / Q000737:chemistry / Q000494:pharmacology*; D013997:Time Factors
chemical_list: D044946:Biflavonoids; D047908:Intracellular Signaling Peptides and Proteins; D047428:Protein Kinase Inhibitors; C092117:RIN1 protein, human; D054852:Small Molecule Libraries; C585473:theaflavin-3,3'-digallate; D002392:Catechin; D016044:Fusion Proteins, bcr-abl; D016315:Proto-Oncogene Proteins c-abl; D019950:Mitogen-Activated Protein Kinase 1