The global rise of life-threatening diseases, particularly breast cancer, has emerged as a grave public health challenge. Recognizing the distinct structural demands of anti-breast cancer targets, we have synthesized Thiophene-DHPMs analogs by scaffold hopping approach to exhibit versatility having the ability to target proteins targets of cancer and their interactions contingent upon the various substitutions on them. We have harnessed MCRto synthesize novel Thiophene-DHPMs (4a-4d) derivatives introducing an efficient Biginelli protocol approach. All derivatives are characterized through techniques including FTIR and 1H NMR and evaluated by in vitro SRB assay method on MCF-7 cell line, compared against positive control ADR. Molecular docking studies against kinesin spindle protein Eg5 (1Q0B) revealed superior binding interactions and docking scores (> 8 Kcal) compared to the prototype Eg5 inhibitor Monastrol. Compound 4a binds via Hydrogen bond interaction to target Eg5 with ALA-133, PRO-137, TYR-211 In vitro evaluation of results indicates that compound 4a found to be moderately active (GI50 = -4.41) compared to positive control ADR (GI50 = -7.76) against MCF-7 cells. Compound 4a demonstrates significant activity due to the presence of R=C2H5, X= S, and thiophene ring.