Zareba_2023_Eur.J.Med.Chem_261_115832

Alzheimer's disease (AD) is a global health problem in the medical sector that will increase over time. The limited treatment of AD leads to the search for a new clinical candidate. Considering the multifactorial nature of AD, a strategy targeting number of regulatory proteins involved in the development of the disease is an effective approach. Here, we present a discovery of new multi-target-directed ligands (MTDLs), purposely designed as GABA transporter (GAT) inhibitors, that successfully provide the inhibitory activity against butyrylcholinesterase (BuChE), beta-secretase (BACE1), amyloid beta aggregation and calcium channel blockade activity. The selected GAT inhibitors, 19c and 22a - N-benzylamide derivatives of 4-aminobutyric acid, displayed the most prominent multifunctional profile. Compound 19c (mGAT1 IC(50) = 10 microM, mGAT4 IC(50) = 12 microM and BuChE IC(50) = 559 nM) possessed the highest hBACE1 and Abeta(40) aggregation inhibitory activity (IC(50) = 1.57 microM and 99 % at 10 microM, respectively). Additionally, it showed a decrease in both the elongation and nucleation constants of the amyloid aggregation process. In contrast compound 22a represented the highest activity and a mixed-type of eqBuChE inhibition (IC(50) = 173 nM) with hBACE1 (IC(50) = 9.42 microM), Abeta aggregation (79 % at 10 microM) and mGATs (mGAT1 IC(50) = 30 microM, mGAT4 IC(50) = 25 microM) inhibitory activity. Performed molecular docking studies described the mode of interactions with GATs and enzymatic targets. In ADMET in vitro studies both compounds showed acceptable metabolic stability and low neurotoxicity. Successfully, compounds 19c and 22a at the dose of 30 mg/kg possessed statistically significant antiamnesic properties in a mouse model of amnesia caused by scopolamine and assessed in the novel object recognition (NOR) task or the passive avoidance (PA) task.