Chemical and computational methods for the characterization of covalent reactive groups for the prospective design of irreversible inhibitors.
Interest in drugs that covalently modify their target is driven by the desire for enhanced efficacy that can result from the silencing of enzymatic activity until protein resynthesis can occur, along with the potential for increased selectivity by targeting uniquely positioned nucleophilic residues in the protein. However, covalent approaches carry additional risk for toxicities or hypersensitivity reactions that can result from covalent modification of unintended targets. Here we describe methods for measuring the reactivity of covalent reactive groups (CRGs) with a biologically relevant nucleophile, glutathione (GSH), along with kinetic data for a broad array of electrophiles. We also describe a computational method for predicting electrophilic reactivity, which taken together can be applied to the prospective design of thiol-reactive covalent inhibitors.
Center of Chemistry Innovation and Excellence, and ‡Department of Pharmacokinetics, Dynamics and Metabolism, Pfizer, Inc. , Eastern Point Road, Groton, Connecticut 06340, United States.
Mark E
ME
Flanagan
Joseph A
JA
Abramite
Dennis P
DP
Anderson
Ann
A
Aulabaugh
Upendra P
UP
Dahal
Adam M
AM
Gilbert
Chao
C
Li
Justin
J
Montgomery
Stacey R
SR
Oppenheimer
Tim
T
Ryder
Brandon P
BP
Schuff
Daniel P
DP
Uccello
Gregory S
GS
Walker
Yan
Y
Wu
Matthew F
MF
Brown
Jinshan M
JM
Chen
Matthew M
MM
Hayward
Mark C
MC
Noe
R Scott
RS
Obach
Laurence
L
Philippe
Veerabahu
V
Shanmugasundaram
Michael J
MJ
Shapiro
Jeremy
J
Starr
Justin
J
Stroh
Ye
Y
Che
D015195:Drug Design; D004791:Enzyme Inhibitors / Q000737:chemistry*; D005978:Glutathione / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D013058:Mass Spectrometry; D019906:Nuclear Magnetic Resonance, Biomolecular; D004364:Pharmaceutical Preparations / Q000737:chemistry
D004791:Enzyme Inhibitors; D004364:Pharmaceutical Preparations; D005978:Glutathione