<p>In this paper, an extensive investigation on the molecular structure, spectroscopic and electronic properties of 3,4',7,8-Tetrahydroxyflavone hydrate (7,8THFH) molecule in the ground state and in the excited state optimization has been employed. FT-IR, FT-Raman, UV–vis, ¹³C and ¹H NMR investigations have been applied by developing DFT computations based on the B3LYP and CAM-B3LYP methods with 6–311 +  + G(d,p) basis set, which also supports providing useful information about the molecular structure of 7,8THFH compound. The vibrational assignments of the title compound were investigated by means of quantum chemical calculations at B3LYP and CAM-B3LYP functional level of theories using 6–311 +  + G(d,p) basis sets, and the results were compared with the experimental data. NMR has been supported by CAM-B3LYP method at 6–311 +  + G(d,p) basis set along with the GIAO approach for assigning ¹³C and ¹H chemical shifts. Over the checkpoint file of the molecular structure, HOMO, LUMO and MEP analyses have been carried out. The quantum chemical reactivities were correlated with the biological properties of the molecule. The electronic charge density concentration in the studied compound was calculated by the Mulliken atomic charges, which aid in determining the molecule to bind in the target receptors. NBO analysis suggested that the existence of hyperconjugative interactions was an essential cause for the chemical stability of the molecule. The physicochemical and <i>in-silico</i> ADMET predictions are used to analyze whether the 7,8THFH drug is a possible therapeutic for breast cancer. Molecular docking analysis suggest that 7,8THFH ligand has a good biological activity with values of –10.51 and –8.92&#xa0;kcal/mol for two different protein receptor 1U3G and 3EQM, respectively. The anti-cancer activity indicates that the compound 7,8THFH needed to inhibit this biological process by half (IC50) was found to be 24.8&#xa0;µg, which may act as a good drug candidate. Lastly, the findings of MMP-Rh-123, propidium iodide and Western-blot assays confirmed that compound 7,8THFH possessed good anticancer activity against MCF-7 cancer cell line. The observed and simulated results are highly correlated, confirming the molecular structure, and the findings of in vitro analyses suggest that 7,8THFH acts as an effective inhibitor against the MCF-7 cell lines, highlighting its strong antiproliferative activity.</p>