Integrated Biochemical and Computational Evaluation of Bis-Imidazolium Derivatives 3e–3g in Cholinesterase and HIV-1 Reverse Transcriptase Assays

Bis-imidazolium derivatives (3e, 3f, 3g) were evaluated for their inhibitory activity against acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT) using colorimetric assays and molecular docking. For the HIV-1 RT–3e complex, a 100 ns molecular dynamics (MD) simulation and Molecular Mechanics/Generalized Born Surface Area (MM/GBSA) analysis were performed. Compound 3e emerged as the most active derivative, showing the lowest half-maximal inhibitory concentration (IC50) values: 13.36 µg/mL for AChE, 4.95 µg/mL for BChE, and 50.44 µg/mL for HIV-1 RT. Nevirapine produced 95.33% RT inhibition at 20 µM. Redocking validated the binding poses with root-mean-square deviation (RMSD) values below 2.0 Å, though docking scores lacked consistent correlation with experimental activity rankings. In the MD trajectory, 3e remained stable within the RT binding pocket, yielding an MM/GBSA energy of −51.04 ± 8.39 kcal/mol. Absorption, distribution, metabolism, excretion, and toxicity (ADMET) predictions indicated low gastrointestinal absorption, no blood–brain barrier permeation, and a high-confidence human ether-à-go-go-related gene (hERG) potassium channel blockade alert. These findings highlight 3e for further optimization.