Pushpa V H Jayanthi M K ^* N Akshaya Simha Siva Dallavalasa Sapna Patel M C Ramith Ramu SubbaRao V. Madhunapantula

• JSS Academy of Higher Education and Research, Mysore, Karnataka, India

Shivagutika, a polyherbal drug mentioned in Ayurveda, has been shown to have anti-breast cancer potential in vitro and in silico using breast cancer cell lines MCF-7, MDA-MB-231, and MDA-MB-468 & Caspase 3 as a target respectively from the previous study. The current study aims to further explore its anti-breast cancer potential under in vivo and in silico conditions with different targets. The current objective of the current study's was to examine in vivo anti-breast cancer potential of Shivagutika Dichloromethane (DCM) extract by utilising an EAC solid tumour model in Swiss albino mice. Six animals were studied for acute oral toxicity, with doses of 300 mg/kg body weight (b.wt.) and 2000 mg/kg body weight (b.wt.). The EAC model was designed, and EAC cells from ascitic fluid of mice were injected into left thigh tissue of swiss albino mice. Anti-tumor assays, physical, biochemical, haematological, histopathological studies and cell cycle studies were performed. Molecular docking and simulation studies were conducted with DCM extract's phytochemicals targeting p53 and Ki67. The DCM extract showed no deaths or behavioural changes in acute oral toxicity studies at concentrations of 300 mg/kg b.wt. and 2000 mg/kg b.wt. At 500 mg/kg b.wt., it showed potent anti-tumor activity, increasing survival time, reducing body weight, and normalizing biochemical parameters. DCM extract also arrested the cell cycle at G2/M and increase in the number of cells in G0/G1 phase of cell cycle. It inhibited Ki67 expression and activated p53. In-silico analysis revealed that Sciadopitysin, as a potent compound, interacted with p53 and Ki67 with binding affinities of -8.4 kcal/mol and -8.0 kcal/mol, respectively by forming stable complexes during simulation.