Objectives: To investigate rotundic acid (RA), a triterpenoid, for its protective role against the cyclophosphamide (CP) induced cardiotoxicity in Swiss albino mice, as CP, although a potent anticancer drug, induces severe cardiotoxicity as a side effect in cancer patients. Thus, this study was a step towards developing an adjuvant that can reduce the toxicity of CP during cancer treatment. Materials and Methods: Animals were randomly assigned to 7 groups. Control; CP 200; RA 10 + CP 200; RA 20 + CP 200; RA 40 + CP 200; NER 400 + CP 200 and RA 40 Results: CP 200 mg/kg, IP, enhanced the level of cardiac troponin T, CK-MB, LDH, NF-κB, TLR4, TNF-α, IL-6, IL-Iβ, cleaved caspase-3, TBARS, nitrite, and reduced the level of CAT, GSH, and SOD, resulting in cardiac injury, nitrative stress, oxidative stress, inflammation, and fibrosis. Administration of RA and nerolidol substantially reversed these pathological modifications to normal. Molecular docking study showed that RA strongly binds to the pocket domains of TLR-4 and cleaves caspase-3. Conclusion: The findings suggest that RA has the potential to reduce the toxicity of CP in the heart and can be utilized as an additional treatment for cancer therapy, along with CP. However, further research using animal models for cancer is required to confirm this.