title: Discovery of novel, dual mechanism ERK inhibitors by affinity selection screening of an inactive kinase.
abstract: An affinity-based mass spectrometry screening technology was used to identify novel binders to both nonphosphorylated and phosphorylated ERK2. Screening of inactive ERK2 identified a pyrrolidine analogue 1 that bound to both nonphosphorylated and phosphorylated ERK2 and inhibited ERK2 kinase activity. Chemical optimization identified compound 4 as a novel, potent, and highly selective ERK1,2 inhibitor which not only demonstrated inhibition of phosphorylation of ERK substrate p90RSK but also demonstrated inhibition of ERK1,2 phosphorylation on the activation loop. X-ray cocrystallography revealed that upon binding of compound 4 to ERK2, Tyr34 undergoes a rotation (flip) along with a shift in the poly-Gly rich loop to create a new binding pocket into which 4 can bind. This new binding mode represents a novel mechanism by which high affinity ATP-competitive compounds may achieve excellent kinase selectivity.
authors.affiliation: Merck Research Laboratories , 33 Avenue Louis Pasteur, Boston, Massachusetts 02115, United States.
authors.forename: Yongqi
authors.initials: Y
authors.lastname: Deng
authors.forename: Gerald W
authors.initials: GW
authors.lastname: Shipps
authors.forename: Alan
authors.initials: A
authors.lastname: Cooper
authors.forename: Jessie M
authors.initials: JM
authors.lastname: English
authors.forename: D Allen
authors.initials: DA
authors.lastname: Annis
authors.forename: Donna
authors.initials: D
authors.lastname: Carr
authors.forename: Yang
authors.initials: Y
authors.lastname: Nan
authors.forename: Tong
authors.initials: T
authors.lastname: Wang
authors.forename: Hugh Y
authors.initials: HY
authors.lastname: Zhu
authors.forename: Cheng-Chi
authors.initials: CC
authors.lastname: Chuang
authors.forename: Priya
authors.initials: P
authors.lastname: Dayananth
authors.forename: Alan W
authors.initials: AW
authors.lastname: Hruza
authors.forename: Li
authors.initials: L
authors.lastname: Xiao
authors.forename: Weihong
authors.initials: W
authors.lastname: Jin
authors.forename: Paul
authors.initials: P
authors.lastname: Kirschmeier
authors.forename: William T
authors.initials: WT
authors.lastname: Windsor
authors.forename: Ahmed A
authors.initials: AA
authors.lastname: Samatar
mesh_terms: D000345:Affinity Labels; D000813:Anilides / Q000378:metabolism* / Q000494:pharmacology; D018360:Crystallography, X-Ray; D020128:Inhibitory Concentration 50; D020935:MAP Kinase Signaling System / Q000187:drug effects*; D013058:Mass Spectrometry / Q000379:methods; D019950:Mitogen-Activated Protein Kinase 1 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors / Q000138:chemical synthesis* / Q000494:pharmacology; D011759:Pyrrolidines / Q000378:metabolism* / Q000494:pharmacology; D013329:Structure-Activity Relationship
chemical_list: C000595643:1-(2-(4-(4-acetylphenyl)piperazin-1-yl)-2-oxoethyl)-N-(3-chloro-4-hydroxyphenyl)pyrrolidine-3-carboxamide; D000345:Affinity Labels; D000813:Anilides; D047428:Protein Kinase Inhibitors; D011759:Pyrrolidines; D019950:Mitogen-Activated Protein Kinase 1