SSB-2548 Attenuates C-X-C chemokine receptor type 4 (CXCR-4) Activation to Induce Apoptosis; An in silico Guided invitro Validation in Acute Myeloid Leukemia cells

Abstract

Background and Aim: The role of C-X-C chemokine receptor type 4 (CXCR-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. Methods Diversity-based virtual screening process using Autodock-vina was employed to screen approximately 8,50,000 compounds from the ChemBridge-small molecule database. The binding stability and dynamics was investigated through GROMACS-based molecular dynamics simulations and RMSD. AML cells (THP-1, HL-60 and SKM-1cell cell lines) was used to assess proliferation CXCR-4 expression and apoptosis induction were measured using flow cytometry and trans-endothelial migration was assessed using calorimetric method in AML cells. The ADME properties were predicted using SwissADME server. Results 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-1cell proliferations with GI50 values of 84.57 nM, 41.30 nM 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. Conclusion Computational assessments identified SSB-2548 as a novel CXCR-4 inhibitor. Invitro 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.