Language

English

Publication Date

1-1-2025

Journal

American Journal of Cancer Research

DOI

10.62347/ARCZ1924

PMID

40948519

PMCID

PMC12432553

PubMedCentral® Posted Date

8-25-2025

PubMedCentral® Full Text Version

Post-print

Abstract

This study explores the synthesis, characterization, and therapeutic efficacy of AGY1, AGY2, AGY3, AGY4, and AGY5, which are novel 5-FU analogs designed to improve metabolic stability, prolong half-life, and anti-tumor activity against pancreatic cancer. The 5-FU molecule was chemically modified to bypass dihydropyrimidine dehydrogenase (DPD)-mediated inactivation, enhancing drug retention and increasing lipophilicity for improved cellular uptake. The analog cytotoxic activity was evaluated in 2D monolayer cultures and 3D pancreatic cancer spheroids and organoid models derived from MiaPaCa-2 and PANC-1 cells to simulate a more complex tumor environment. In the 2D model, AGY1, AGY2, AGY3, AGY4, and AGY5 displayed significantly higher cytotoxicity than 5-FU, with AGY2 achieving up to six-fold higher potency in MiaPaCa-2 cells. In 3D spheroid models, both AGY1 and AGY2 showed dose-dependent reductions in spheroid size, with AGY2 causing the most pronounced shrinkage, suggesting effective disruption of the tumor architecture. In pancreatic organoids, AGY2 demonstrated substantial decreases in cell viability and structural proliferation, inhibiting cell migration and organoid budding that exceeded the effects of 5-FU. Furthermore, cell cycle analysis revealed that AGY2 induces significant cell cycle arrest at the G0/G1 phase in MiaPaCa-2 cells and the S phase in PANC-1 cells. Apoptosis assays showed a higher percentage of apoptotic cells following AGY2 treatment compared to 5-FU, which was supported by Western blot analysis, indicating increased expression of pro-apoptotic proteins p53 and Bax and decreased levels of survival proteins epidermal growth factor receptor (EGFR), human epidermal growth factor receptor 2 (HER-2), and Poly (ADP-ribose) polymerase (PARP). Put together, our findings showed that AGY2 analog was the most effective anti-anticancer analog with significantly improved metabolic stability.

Keywords

Pancreatic cancer, 5-Fluorouracil, analogs, 2D and 3D cell models, apoptosis, Western blot

Published Open-Access

yes

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