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ABSTRACT Ovarian cancer (OvCa) remains the leading cause of gynecological cancer mortality, with most patients developing chemoresistance. Drug repurposing offers promising alternatives, with mebendazole (MBZ) showing anticancer activity. This study evaluates MBZ efficacy using Spectral Domain Optical Coherence Tomography (SD‐OCT). We conducted longitudinal imaging of 40 wild‐type (WT) and cisplatin‐resistant (CPR) OVCAR8 multicellular tumor spheroids over 11 days. Four analyses were performed: volume analysis, optical attenuation analysis, uniformity analysis, and texture feature analysis. Volume analysis showed MBZ reduced spheroid growth in both groups, with greater effects in CPR‐MCTs. Optical attenuation analysis revealed increased necrotic tissue ratios in treated spheroids. Uniformity analysis demonstrated MBZ targets heterogeneous tissues effectively. Texture analysis identified significant structural changes, with 866 altered features in CPR spheroids versus 124 in WT spheroids. Cell viability assays confirmed MBZ's effectiveness against standard and chemo‐resistant OVCAR8 tumors. This study demonstrates SD‐OCT's utility for noninvasive therapy monitoring in 3D cancer models.