Characterization of C-alkyl amidines as bioavailable covalent reversible inhibitors of human DDAH-1.

ABSTRACT: C-Alkyl amidine analogues of asymmetric N(?),N(?)-dimethyl-L-arginine are dual-targeted inhibitors of both human DDAH-1 and nitric oxide (NO) synthase, and provide a promising scaffold for the development of therapeutics to control NO overproduction in a variety of pathologies including septic shock and some cancers. Using a two-part click-chemistry-mediated activity probe, a homologated series of C-alkyl amidines were ranked for their ability to inhibit DDAH-1 within cultured HEK 293T cells. N?-(1-Iminopentyl)-L-ornithine was determined to be the most potent compound in?vitro (K(d)=7??M) as well as in cultured cells, and the binding conformation and covalent reversible mode of inhibition was investigated by comparison of interactions made with DDAH-1 and a catalytically inactive C274S variant, as gauged by X-ray crystallography and isothermal titration calorimetry. By interrupting the ability of the inhibitor to form a covalent bond, the contribution of this interaction could be estimated. These results suggest that further stabilization of the covalent adduct is a promising strategy for lead optimization in the design of effective reagents to block NO synthesis.

SUBMITTER: Lluis M 

PROVIDER: S-EPMC3251910 | biostudies-literature | 2011 Jan

REPOSITORIES: biostudies-literature