Four thiosemicarbazones by using tolualdehyde and cuminaldehyde having the formula, R2-(S) C-HN-N--HC-R1[R1 = CH3 & R2 = C4H9N (TAP), R2 = C4H9NO & R1 = CH3 & (TAM), R2 = C4H9N & R1 = (CH3)2CH (CMP), R2 = C4H9NO & R1 = (CH3)2CH (CMM)] have been synthesized. The compound interactions were assessed using their UV-visible, infrared, NMR, and HRMS spectra. Single-crystal X-ray diffraction was employed to know the molecular structure of CMP and TAP. The compounds were assessed for their interactions with Calf-Thymus (CT)-DNA using spectroscopic titrations using both emission and absorption spectra. As per the research findings on DNA binding, the compounds interactively interacted with DNA, as indicated by the hypochromic and slight red shift. TAP exhibited a high binding constant (5.16 pound 105), suggesting a stronger binding to CT-DNA compared to other compounds. The fluorescence titration spectra of BSA binding experiments exhibited a noteworthy hypochromic shift and red shift, displaying a strong interaction of chemicals with BSA. EGFR protein docking examination demonstrated the potential of compounds to treat the targets. TAP displayed the highest binding score (-6.4230 Kcal/mol) to EGFR with the four compounds. To compute density functional theory (DFT), B3LYP/6-311 G (d, p) level theories have been implemented. Generated compounds' computational analyses reveal the structural stability of compound TAP than the rest synthesized ligands. SwissADME investigations indicate that the LogP values for each compound are less than five indicating that they have the right lipophilicity characteristics. All newly synthesized compounds follow Lipinski's rule of drug lines. A good result for this characteristic is indicated by the low degree of synthetic accessibility, which falls between two and three. Each of the compounds (TAP-CMM) can develop into a viable oral medicine. Each compound (TAP-CMM) was tested for its anticancer potential using MCF-7, MCF-10A, A549, and human HepG-2 liver. TAP demonstrated favorable efficacy in HepG-2 liver cancer cells, exhibiting IC50 values of 23.1 mu M. C1 [Singh, Vipin; Sreekanth, Anandaram] Natl Inst Technol, Dept Chem, Tiruchirappalli 620015, Tamil Nadu, India. [Haribabu, Jebiti] Univ Atacama, Fac Med, Los Carreras 1579, Copiapo 1532502, Chile. [Haribabu, Jebiti] Chennai Inst Technol CIT, Chennai 600069, Tamil Nadu, India. [Moraga, Daniel] Univ Tarapaca, Fac Med, Dept Ciencias Biomed, Lab Fisiol, Arica 1000000, Chile. [Santibanez, Juan F.] Univ Belgrade, Inst Med Res, Natl Inst Republ Serbia, Belgrade, Serbia. [Santibanez, Juan F.] Bernardo OHiggins Univ, Integrat Ctr Biol & Appl Chem CIBQA, Santiago, Chile. C3 National Institute of Technology (NIT System); National Institute of Technology Tiruchirappalli; Universidad de Atacama; Universidad de Tarapaca; University of Belgrade; Universidad Bernardo O'Higgins