20.500.12556/RUL-179045 Expanding the reactive chemistry toolbox – warhead screening for covalent butyrylcholinesterase inhibitors Many covalent inhibitors of butyryl- and acetylcholinesterase, typically featuring electrophilic carbamoyl, phosphonyl, and sulfonyl groups, were reported in the literature. In this study, we screened these and several other electrophilic moieties for covalent cholinesterase inhibition. The electrophilic warheads were either installed at different positions on a reversible, selective human butyrylcholinesterase inhibitor scaffold or featured as small molecular weight, fragment-sized compounds. Time-dependency of the enzyme inhibition served as an indicator of covalent binding. The 7-indolyl substitution pattern proved optimal for generating time-dependent inhibitors, and carbamate chemotype produced most of the time-dependent hit compounds. Interestingly, the carbamates' reactivity was strongly influenced by their leaving groups’ nucleofugality and not by steric hindrance, as sufficiently acidic leaving groups (pKa <10) enabled carbamoylation of the catalytic serine. Notably, an exception to this rule of a thumb was found in a series of chalcogen carbachol analogues. The gathered insights highlight some key structure–reactivity relationships for covalent hBChE inhibitors and may assist in development of novel serine hydrolase inhibitors. cholinesterase inhibitors warhead irreversible inhibition butyrylcholinesterase carbamate zaviralci holinesteraze bojna glava ireverzibilna inhibicija butirilholinesteraza karbamat farmacevtska kemija analizna kemija true false true Angleški jezik Slovenski jezik Članek v reviji 2026-02-04 08:30:53 2026-02-04 08:30:57 2026-06-18 11:54:55 0000-00-00 00:00:00 2026 0 0 26 str. art. 111841 Vol. 423 Jan. 2026 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 615.4:54:616.894 1872-7786 10.1016/j.cbi.2025.111841 260611587 1-s2.0-S0009279725004715-main.pdf 1-s2.0-S0009279725004715-main.pdf 1 C21F3AA0C2E550E36873308A0CFDB757 f24b80ac22e87d1881d6e5c723b585b30f1445e667fddbcc3ebbbedb852e6723 bd5ec693-019b-11f1-a1ba-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=227146 https://www.sciencedirect.com/science/article/pii/S0009279725004715 1 5cde68f0-019b-11f1-a1ba-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=227145 Fakulteta za farmacijo Medicinska fakulteta 0 0 0