Design and Development of a Chemical Probe for Pseudokinase Ca2+/calmodulin-Dependent Ser/Thr Kinase

CASK (Ca2+/calmodulin-dependent Ser/Thr kinase) is a member of the MAGUK (membrane-associated guanylate kinase) family that functions as neurexin kinases with roles implicated in neuronal synapses and trafficking. The lack of a canonical DFG motif, which is altered to GFG in CASK, led to the classification as a pseudokinase. However, functional studies revealed that CASK can still phosphorylate substrates in the absence of divalent metals. CASK dysfunction has been linked to many diseases, including colorectal cancer, Parkinson's disease, and X-linked mental retardation, suggesting CASK as a potential drug target. Here, we exploited structure-based design for the development of highly potent and selective CASK inhibitors based on 2,4-diaminopyrimidine-5-carboxamides targeting an unusual pocket created by the GFG motif. The presented inhibitor design offers a more general strategy for the development of pseudokinase ligands that harbor unusual sequence motifs. It also provides a first chemical probe for studying the biological roles of CASK.

Automated summary

CASK, a member of the MAGUK family, functions as a neurexin kinase and is classified as a pseudokinase due to the lack of a canonical DFG motif. Despite this classification, CASK has been found to phosphorylate substrates in the absence of divalent metals. Its dysfunction has been linked to various diseases, making it a potential drug target. The development of highly potent and selective CASK inhibitors based on an unusual pocket created by the GFG motif offers a general strategy for developing pseudokinase ligands and provides a chemical probe for studying the biological roles of CASK.