title: Development of Novel In Vivo Chemical Probes to Address CNS Protein Kinase Involvement in Synaptic Dysfunction.
abstract: Serine-threonine protein kinases are critical to CNS function, yet there is a dearth of highly selective, CNS-active kinase inhibitors for in vivo investigations. Further, prevailing assumptions raise concerns about whether single kinase inhibitors can show in vivo efficacy for CNS pathologies, and debates over viable approaches to the development of safe and efficacious kinase inhibitors are unsettled. It is critical, therefore, that these scientific challenges be addressed in order to test hypotheses about protein kinases in neuropathology progression and the potential for in vivo modulation of their catalytic activity. Identification of molecular targets whose in vivo modulation can attenuate synaptic dysfunction would provide a foundation for future disease-modifying therapeutic development as well as insight into cellular mechanisms. Clinical and preclinical studies suggest a critical link between synaptic dysfunction in neurodegenerative disorders and the activation of p38αMAPK mediated signaling cascades. Activation in both neurons and glia also offers the unusual potential to generate enhanced responses through targeting a single kinase in two distinct cell types involved in pathology progression. However, target validation has been limited by lack of highly selective inhibitors amenable to in vivo use in the CNS. Therefore, we employed high-resolution co-crystallography and pharmacoinformatics to design and develop a novel synthetic, active site targeted, CNS-active, p38αMAPK inhibitor (MW108). Selectivity was demonstrated by large-scale kinome screens, functional GPCR agonist and antagonist analyses of off-target potential, and evaluation of cellular target engagement. In vitro and in vivo assays demonstrated that MW108 ameliorates beta-amyloid induced synaptic and cognitive dysfunction. A serendipitous discovery during co-crystallographic analyses revised prevailing models about active site targeting of inhibitors, providing insights that will facilitate future kinase inhibitor design. Overall, our studies deliver highly selective in vivo probes appropriate for CNS investigations and demonstrate that modulation of p38αMAPK activity can attenuate synaptic dysfunction.
authors.affiliation: Department of Molecular Pharmacology and Biological Chemistry, Northwestern University, Chicago, Illinois, United States of America.
authors.forename: D Martin
authors.initials: DM
authors.lastname: Watterson
authors.forename: Valerie L
authors.initials: VL
authors.lastname: Grum-Tokars
authors.forename: Saktimayee M
authors.initials: SM
authors.lastname: Roy
authors.forename: James P
authors.initials: JP
authors.lastname: Schavocky
authors.forename: Brinda Desai
authors.initials: BD
authors.lastname: Bradaric
authors.forename: Adam D
authors.initials: AD
authors.lastname: Bachstetter
authors.forename: Bin
authors.initials: B
authors.lastname: Xing
authors.forename: Edgardo
authors.initials: E
authors.lastname: Dimayuga
authors.forename: Faisal
authors.initials: F
authors.lastname: Saeed
authors.forename: Hong
authors.initials: H
authors.lastname: Zhang
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Staniszewski
authors.forename: Jeffrey C
authors.initials: JC
authors.lastname: Pelletier
authors.forename: George
authors.initials: G
authors.lastname: Minasov
authors.forename: Wayne F
authors.initials: WF
authors.lastname: Anderson
authors.forename: Ottavio
authors.initials: O
authors.lastname: Arancio
authors.forename: Linda J
authors.initials: LJ
authors.lastname: Van Eldik
mesh_terms: D016229:Amyloid beta-Peptides / Q000633:toxicity; D000818:Animals; D001921:Brain / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism; D020134:Catalytic Domain; D002460:Cell Line; D015195:Drug Design; D006801:Humans; D017774:Long-Term Potentiation / Q000187:drug effects; D008297:Male; D008810:Mice, Inbred C57BL; D048308:Mitogen-Activated Protein Kinase 14 / Q000037:antagonists & inhibitors* / Q000737:chemistry; D008958:Models, Molecular; D010446:Peptide Fragments / Q000633:toxicity; D047428:Protein Kinase Inhibitors / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology*; D011724:Pyridazines / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology*; D011725:Pyridines / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology*
chemical_list: D016229:Amyloid beta-Peptides; C000608102:N,N-dimethyl-6-(naphthalen-2-yl)-5-(pyridin-4-yl)pyridazin-3-amine; D010446:Peptide Fragments; D047428:Protein Kinase Inhibitors; D011724:Pyridazines; D011725:Pyridines; C075222:amyloid beta-protein (1-42); D048308:Mitogen-Activated Protein Kinase 14