Unraveling the Therapeutic Potential of a Novel Proquazone Analog (PA-6) to Harness the Anti Leukemia Mechanisms in K562 Cell Lines by Proteomics and Systems Biology Approaches

Leukemia is an aggressive hematologic malignancy that necessitates safer therapies due to the adverse effects of existing treatments. In this study, we evaluated the anti-leukemic potential of PA-6, a Proquazone analog (6-amino-1-isopropyl-7-methyl-4-phenylquinazolin-2(1H)-one), using a comprehensive systems biology approach and global proteomic analysis in K562 leukemia cells via Orbitrap mass spectrometry. PA-6 was synthesized and structurally confirmed through NMR and HRMS (ESI-TOF) analysis. Computational profiling, including pharmacokinetic and molecular docking studies, was performed before treating K562 cells with PA-6 (5/20µM, 48-72h). Proteomic analysis involved protein extraction, digestion, peptide purification, and Orbitrap MS/MS-based identification of differentially expressed proteins, followed by functional annotation. PA-6 exhibited favorable drug-likeness properties, such as optimal molecular weight, absorption, and blood-brain barrier permeability, indicating its potential as a therapeutic agent. Molecular docking identified MTA2 and HNRNPM proteins, which regulate RNA metabolism and chromatin remodeling, as potential targets of PA-6. In-vitro validation via cytotoxicity assays revealed dose-dependent suppression of key signaling pathways, leading to a significant reduction in K562 cell proliferation. Proteomic analysis further demonstrated alterations in apoptosis, RNA processing, and cell cycle regulation pathways, highlighting its molecular and cellular impact. Overall, this study provides valuable insights into PA-6's anti-leukemic potential, emphasizing its ability to disrupt crucial cellular and signaling processes in leukemia cells. Its promising pharmacokinetics, cell proliferation inhibition, and low toxicity suggest that PA-6 could serve as a potential therapeutic candidate for leukemia treatment.