Pregnenolone carbamate derivatives as selective cholinesterase inhibitors: a comprehensive evaluation exploring their interactions with DNA and human serum albumin.

Abstract

Pregnenolone-carbamate derivatives were synthesized and evaluated as potential multifunctional agents with cholinesterase inhibitory and neuroprotective properties. Among them, P3, P5, and P9 exhibited the most promising profiles and were subjected to integrated spectroscopic, biochemical, and computational analyses. Inhibition assays revealed selective acetylcholinesterase (AChE) inhibition by P3 (IC<inf>50</inf> = 0.11 μM), butyrylcholinesterase inhibition (BuChE) selectivity for P5 (IC<inf>50</inf> = 0.47 μM), and dual inhibition by P9. Fluorescence and UV–vis studies indicated minor groove binding to DNA with log K<inf>app</inf> values around 5.85, while static quenching with human serum albumin (HSA) was observed, with log K<inf>A</inf> values up to 2.25. Docking studies supported these findings, showing favorable binding energies for DNA (–8.6 kcal/mol) and HSA (–9.7 kcal/mol), and predicted localization within the DNA minor groove and Sudlow sites on HSA. Cytotoxicity assays on HT22 cells indicated high viability (> 75% at 40 µM), suggesting a favorable safety margin. These results offer a molecular-level understanding of the pharmacodynamic and pharmacokinetic properties of these compounds and support their potential for further in vivo evaluation.