Discovery of Two Novel Pyrazole Derivatives as Anticancer Agents Targeting Tubulin Polymerization and MAPK Signaling Pathways.

Objectives: Drug resistance is the major determinant of chemotherapy failure, leading to relapse and tumor progression, demonstrating the urgent need for novel antineoplastic drugs. This study aimed to evaluate the anticancer potential of two novel pyrazole derivatives, P3C.1 and P3C.2, and to elucidate their mechanism of action in cancer cells. Methods: The cytotoxicity of the compounds was evaluated across 27 different cancer cell lines via a nuclear staining assay. Subsequent flow cytometric and biochemical analyses were performed to assess reactive oxygen species (ROS) generation, apoptosis induction, mitochondrial integrity, and cell cycle progression. Additional studies included transcriptome analyses and immunoassays to characterize the molecular mechanisms underlying drug activity. Results: Two novel pyrazole derivatives, P3C.1 and P3C.2, were identified with potent cytotoxicity on a variety of cancer cell lines. Among the adherent cell lines tested, the triple-negative breast cancer (TNBC) cell line MDA-MB-231 exhibited the highest sensitivity to both compounds and was therefore selected for further experimentation. Conclusions: P3C.1 and P3C.2 emerged as promising anti-breast cancer agents with dual mechanisms of action involving microtubule disruption and altered kinase signaling, leading to induction of apoptosis.