Novel statin derivatives as non-competitive inhibitors of human adenylate kinase 1: synthesis, enzyme inhibition, molecular docking, and biological studies.

Adenylate kinases (AKs) are a family of conserved phosphotransferases essential for maintaining cellular energy homeostasis through the interconversion of adenine nucleotides. AKs also contribute to diverse pathophysiological processes beyond energy metabolism, particularly in inflammation, cardiovascular diseases, and neurodegeneration. Despite their emerging therapeutic relevance, efforts to develop selective AK inhibitors remain limited. Existing inhibitors, such as dinucleoside polyphosphates (e.g., Ap5A), act competitively but display low selectivity, limited bioavailability, and potential off-target effects due to their structural similarity to natural nucleotides. These limitations highlight the need for novel AK inhibitors with improved pharmacological profiles and alternative mechanisms of action. In this study, we synthesized novel statin derivatives and confirmed their non-competitive inhibition of human adenylate kinase 1 (hAK1). Molecular docking and molecular dynamics simulations elucidate the stable binding within the LID domain, providing a structural rationale for the mechanism of non-competitive inhibition. While the derivatives showed micromolar potency against hAK1, they demonstrated a favorable safety and selectivity profile. The effects of statin derivatives on HMG-CoA reductase activity were assessed, showing markedly weaker inhibition than pravastatin, underscoring their enhanced selectivity toward hAK1. Fluorescence spectroscopy revealed strong binding affinities to human and bovine serum albumin. Furthermore, their impact on HDL uptake was evaluated in the human hepatocellular carcinoma cell line (HepG2), while potential cytotoxicity and potency to inhibit membrane-bound adenylate kinase were assessed across multiple cellular models, including cancer (HepG2, A549, SH-SY5Y) and non-cancerous (HUVEC) cells. These findings establish these derivatives as promising scaffolds for the development of selective, non-competitive AK1 inhibitors.