2. Academic Validation
  3. Discovery of a Potent and Selective CDKL5/GSK3 Chemical Probe That Is Neuroprotective

Discovery of a Potent and Selective CDKL5/GSK3 Chemical Probe That Is Neuroprotective

  • ACS Chem Neurosci. 2023 May 3;14(9):1672-1685. doi: 10.1021/acschemneuro.3c00135.
Han Wee Ong 1,  Yi Liang 1,  William Richardson 2,  Emily R Lowry 3 4,  Carrow I Wells 1,  Xiangrong Chen 2,  Margaux Silvestre 5,  Kelvin Dempster 5,  Josie A Silvaroli 6,  Jeffery L Smith 1,  Hynek Wichterle 3 4 7 8 9,  Navjot S Pabla 6,  Sila K Ultanir 5,  Alex N Bullock 2,  David H Drewry 1 10,  Alison D Axtman 1
Affiliations

Affiliations

  • 1 Structural Genomics Consortium, UNC Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, United States.
  • 2 Centre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford, Oxford OX3 7FZ, U.K.
  • 3 Department of Pathology and Cell Biology, Columbia University Irving Medical Center, New York, New York 10032, United States.
  • 4 The Project ALS Therapeutics Core, Columbia University Irving Medical Center, New York, New York 10032, United States.
  • 5 Kinases and Brain Development Laboratory, The Francis Crick Institute, London NW1 1AT, U.K.
  • 6 Division of Pharmaceutics and Pharmacology, College of Pharmacy and Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio 43210, United States.
  • 7 Departments of Neurology, Neuroscience, Rehabilitation and Regenerative Medicine, Columbia University Irving Medical Center, New York, New York 10032, United States.
  • 8 Center for Motor Neuron Biology and Disease, Columbia University Irving Medical Center, New York, New York 10032, United States.
  • 9 Columbia Stem Cell Initiative, Columbia University Irving Medical Center, New York, New York 10032, United States.
  • 10 UNC Lineberger Comprehensive Cancer Center, School of Medicine, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, United States.
PMID: 37084253 DOI: 10.1021/acschemneuro.3c00135
Abstract

Despite mediating several essential processes in the brain, including during development, cyclin-dependent kinase-like 5 (CDKL5) remains a poorly characterized human protein kinase. Accordingly, its substrates, functions, and regulatory mechanisms have not been fully described. We realized that availability of a potent and selective small molecule probe targeting CDKL5 could enable illumination of its roles in normal development as well as in diseases where it has become aberrant due to mutation. We prepared analogs of AT-7519, a compound that has advanced to phase II clinical trials and is a known inhibitor of several cyclin-dependent kinases (CDKs) and cyclin-dependent kinase-like kinases (CDKLs). We identified analog 2 as a highly potent and cell-active chemical probe for CDKL5/GSK3 (glycogen synthase kinase 3). Evaluation of its kinome-wide selectivity confirmed that analog 2 demonstrates excellent selectivity and only retains GSK3α/β affinity. We next demonstrated the inhibition of downstream CDKL5 and GSK3α/β signaling and solved a co-crystal structure of analog 2 bound to human CDKL5. A structurally similar analog (4) proved to lack CDKL5 affinity and maintain potent and selective inhibition of GSK3α/β, making it a suitable negative control. Finally, we used our chemical probe pair (2 and 4) to demonstrate that inhibition of CDKL5 and/or GSK3α/β promotes the survival of human motor neurons exposed to endoplasmic reticulum stress. We have demonstrated a neuroprotective phenotype elicited by our chemical probe pair and exemplified the utility of our compounds to characterize the role of CDKL5/GSK3 in neurons and beyond.

Keywords

CDKL5; GSK3α; GSK3β; chemical probe; crystal structure; kinase; neuroprotective.

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