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[Preprint]. 2025 Oct 23:2025.10.22.683941. [Version 2] doi: 10.1101/2025.10.22.683941

Identification and Validation of an inhibitor of the protein kinases PIM and DYRK

Gyula Bencze, Prabhadevi Venkataramani, Elad Elkayam, Keith D Rivera, Ankur Garg, Istvan Szabadkai, Laszlo Orfi, Leemor Joshua-Tor, Darryl Pappin, Nicholas K Tonks
PMCID: PMC12633467  PMID: 41279372

ABSTRACT

Fermented wheat germ extract (FWGE), a nutraceutical with reported anticancer properties, contains numerous biologically active molecules, but its precise therapeutic constituents remain unclear. In this study, we identify and characterize a novel small-molecule inhibitor, CSH-4044, isolated from FWGE. Through preparative HPLC and structural elucidation via X-ray crystallography, CSH-4044 was revealed to be a unique benzothiazole compound. Kinase profiling demonstrated its high specificity toward the PIM and DYRK families of protein kinases. We determined the co-crystal structure of CSH-4044 bound to PIM1, revealing ATP-competitive binding, and critical hydrophobic and hydrogen-bonding interactions. A chemically synthesized version of CSH-4044 mirrored the activity of the natural product, confirming structural integrity and biological equivalence. Functionally, CSH-4044 suppressed PIM3-driven BAD phosphorylation in pancreatic cancer cells and reduced DYRK1A-mediated Tau phosphorylation in neuronal cells. Our findings position CSH-4044 as a promising lead for targeting PIM and DYRK families of kinases and highlight FWGE as a source of potential therapeutic compounds.

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