Towards the development of selective amine oxidase inhibitors. Mechanism-based inhibition of six copper containing amine oxidases.
Four substrate analogs, 4-(2-naphthyloxy)-2-butyn-1-amine (1), 1,4-diamino-2-chloro-2-butene (2), 1,6-diamino-2,4-hexadiyne (3), and 2-chloro-5-phthalimidopentylamine (4) have been tested as inhibitors against mammalian, plant, bacterial, and fungal copper-containing amine oxidases: bovine plasma amine oxidase (BPAO), equine plasma amine oxidase (EPAO), pea seedling amine oxidase (PSAO), Arthrobacter globiformis amine oxidase (AGAO), Escherichia coli amine oxidase (ECAO), and Pichia pastoris lysyl oxidase (PPLO). Reactions of 1,4-diamino-2-butyne with selected amine oxidases were also examined. Each substrate analog contains a functional group that chemical precedent suggests could produce mechanism-based inactivation. Striking differences in selectivity and rates of inactivation were observed. For example, between two closely related plasma enzymes, BPAO is more sensitive than EPAO to 1 and 3, while the reverse is true for 2 and 4. In general, inactivation appears to arise in some cases from TPQ cofactor modification and in other cases from alkylation of protein residues in a manner that blocks access of substrate to the active site. Notably, 1 completely inhibits AGAO at stoichiometric concentrations and is not a substrate, but is an excellent substrate of PSAO and inhibition is observed only at very high concentrations. Structural models of 1 in Schiff base linkage to the TPQ cofactor in AGAO and PSAO (for which crystal structures are available) reveal substantial differences in the degree of interaction of bound 1 with side-chain residues, consistent with the widely divergent activities. Collectively, these results suggest that the development of highly selective amine oxidase inhibitors is feasible.
Shepard
Eric M
EM
Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT 59717, USA.
Smith
Jennifer
J
Elmore
Bradley O
BO
Kuchar
Jason A
JA
Sayre
Lawrence M
LM
Dooley
David M
DM
D006631:Amine Oxidase (Copper-Containing) / Q000037:antagonists & inhibitors* / Q000737:chemistry; D000818:Animals; D001419:Bacteria / Q000201:enzymology; D003959:Diamines / Q000494:pharmacology; D004295:Dihydroxyphenylalanine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D015195:Drug Design; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D008322:Mammals; D008958:Models, Molecular; D009281:Naphthalenes / Q000737:chemistry / Q000494:pharmacology*; D010659:Phenylhydrazines / Q000737:chemistry; D011487:Protein Conformation; D008249:Protein-Lysine 6-Oxidase / Q000037:antagonists & inhibitors / Q000737:chemistry; D013045:Species Specificity; D013997:Time Factors; D017186:Titrimetry
C086758:2,4,5-trihydroxyphenylalanine quinone; C464879:4-(2-naphthyloxy)-2-butyn-1-amine; D003959:Diamines; D004791:Enzyme Inhibitors; D009281:Naphthalenes; D010659:Phenylhydrazines; C030299:phenylhydrazine; C063335:1,4-diamino-2-butyne; D004295:Dihydroxyphenylalanine; D008249:Protein-Lysine 6-Oxidase; D006631:Amine Oxidase (Copper-Containing)