title: Amide-modified prenylcysteine based Icmt inhibitors: Structure-activity relationships, kinetic analysis and cellular characterization.
abstract: Human protein isoprenylcysteine carboxyl methyltransferase (hIcmt) is the enzyme responsible for the α-carboxyl methylation of the C-terminal isoprenylated cysteine of CaaX proteins, including Ras proteins. This specific posttranslational methylation event has been shown to be important for cellular transformation by oncogenic Ras isoforms. This finding led to interest in hIcmt inhibitors as potential anti-cancer agents. Previous analog studies based on N-acetyl-S-farnesylcysteine identified two prenylcysteine-based low micromolar inhibitors (1a and 1b) of hIcmt, each bearing a phenoxyphenyl amide modification. In this study, a focused library of analogs of 1a and 1b was synthesized and screened versus hIcmt, delineating structural features important for inhibition. Kinetic characterization of the most potent analogs 1a and 1b established that both inhibitors exhibited mixed-mode inhibition and that the competitive component predominated. Using the Cheng-Prusoff method, the K(i) values were determined from the IC(50) values. Analog 1a has a K(IC) of 1.4±0.2μM and a K(IU) of 4.8±0.5μM while 1b has a K(IC) of 0.5±0.07μM and a K(IU) of 1.9±0.2μM. Cellular evaluation of 1b revealed that it alters the subcellular localization of GFP-KRas, and also inhibits both Ras activation and Erk phosphorylation in Jurkat cells.
authors.affiliation: Department of Medicinal Chemistry and Molecular Pharmacology, Purdue University, West Lafayette, IN 47907, USA.
authors.forename: Jaimeen D
authors.initials: JD
authors.lastname: Majmudar
authors.forename: Heather B
authors.initials: HB
authors.lastname: Hodges-Loaiza
authors.forename: Kalub
authors.initials: K
authors.lastname: Hahne
authors.forename: James L
authors.initials: JL
authors.lastname: Donelson
authors.forename: Jiao
authors.initials: J
authors.lastname: Song
authors.forename: Liza
authors.initials: L
authors.lastname: Shrestha
authors.forename: Marietta L
authors.initials: ML
authors.lastname: Harrison
authors.forename: Christine A
authors.initials: CA
authors.lastname: Hrycyna
authors.forename: Richard A
authors.initials: RA
authors.lastname: Gibbs
mesh_terms: D000577:Amides / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D003545:Cysteine / Q000031:analogs & derivatives* / Q000737:chemistry; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology*; D006801:Humans; D019169:Jurkat Cells; D007700:Kinetics; D010766:Phosphorylation; D011496:Protein Methyltransferases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D013329:Structure-Activity Relationship; D018631:ras Proteins / Q000378:metabolism
chemical_list: D000577:Amides; D004791:Enzyme Inhibitors; C542732:prenylcysteine; D011496:Protein Methyltransferases; C056501:protein-S-isoprenylcysteine O-methyltransferase; D018631:ras Proteins; D003545:Cysteine