Synthesis, Characterization and Cytotoxic Effect of a Novel Quinone-based Compound for Cholangiocarcinoma: a Promising Proapoptotic Agent Through Mitochondria

Cholangiocarcinoma patients remain in need of novel therapeutic options due to poor survival. Therefore, a new naphthoquinone derivative, ZP-4, was explored for its cytotoxic potential on CCA cell lines CCLP1 and HUCCT1 after its synthesis and structural characterization. ZP-4 exerted dose-dependent antiproliferative and cytotoxic effects in both cell lines, with CCLP1 cells displaying greater sensitivity than HUCCT1 cells. The reduction in cell viability was accompanied by apoptotic cell death evidenced by pyknotic nuclei and phosphatidylserine translocation. Flow cytometric analyses revealed increased oxidative stress and DNA damage following ZP-4 treatment, supporting a potential association of these cellular responses with the observed apoptotic phenotype. qPCR analysis revealed the upregulation of the pro-apoptotic gene HRK , the stress-responsive gene GADD45A , and the mitophagy-associated gene BNIP3L , further supporting the induction of cellular stress responses by ZP-4. Collectively, these findings identify ZP-4 as a promising candidate for cholangiocarcinoma therapy and support further preclinical evaluation in animal models.