SSB-2548 Inhibits CXCR-4 Activation, Inducing Apoptosis in Acute Myeloid Leukemia Cells

The role of C-X-C Chemokine Receptor type 4 (CXR-4) in chemotherapy resistance remains crucial in promoting proliferation, invasion, and progression in acute myeloid leukemia (AML) cells. This study aims to screen and investigate a potential lead candidate as a therapeutic agent targeting CXCR-4 in AML cells. Diversity-based virtual screening process using AutoDock-Vina was employed to screen approximately 850,000 compounds from the ChemBridge-small molecule database. The binding stability and dynamics were investigated through GROMACS-based molecular dynamics simulations and root mean square deviation (RMSD). AML cells (THP-1, HL-60, and SKM-1 cell lines) were used to assess proliferation CXCR-4 expression, and Apoptosis induction was measured using flow cytometry and trans-endothelial migration was assessed using calorimetric method in AML cells. The absorption, distribution, metabolism, and excretion (ADME) properties were predicted using SwissADME server. The computational evaluations revealed SSB-2548 as a lead candidate that binds stably to CXCR-4. Molecular dynamics simulations provided detailed insights into the conformational changes of the SSB-2548/CXCR-4 complex. The compound inhibited the THP-1, HL-60, and SKM-1 cell proliferations with GI₅₀ values of 84.57, 41.30, and 120.50 nM, respectively. SSB-2548 decreased the trans-endothelial migration and CXCR-4 expression in while inducing early and late phase Apoptosis in all three AML cell types. ADME predictions indicated a favorable lead-likeness, gastrointestinal absorption, and lack of notable toxicity. Computational assessments identified SSB-2548 as a novel CXCR-4 inhibitor. In vitro evaluations proved this lead compound effective against AML cells. These findings lay the groundwork for future, investigations positioning SSB-2548 as a candidate for the development of targeted therapies against AML.

ADME predictions; Acute Myeloid Leukemia (AML); CXCR‐4; apoptosis; computational high throughput screening; molecular dynamic simulation.