Abstract 1823: In vitro evaluation of novel pyrimidine nucleosides for the treatment of pancreatic cancer

Abstract

Abstract The purpose of the study is to synthesize and characterize novel 5-FU analogs (AGY1 and AGY2) and investigate their anticancer activity against pancreatic cancer cell lines. Methods: 5-FU derivatives were synthesized and characterized using Nuclear magnetic resonance (NMR), high-performance liquid chromatography (HPLC), and elemental analysis to determine the presence of new bond formation and the purity of the derivatives or analogs. 2D cell viability, 3D models, cell migration, and cell cycle studies were performed on MiaPaCa-2 (MP-2) cells and PANC-1 cells. Apoptosis was performed on PANC-1 cells by staining with Acridine Orange and Ethidium Bromide. The metabolic stability of the analogs was determined by using human liver microsomes. PARP, p53, and BAX expressions in PANC-1 were determined by Western Blot. Results: NMR data showed the conjugation of alkyl and tetrahydrofuran groups to the 5-FU ring confirming the formation of AGY1 and AGY2. The elemental analysis of AGY1 and AGY2 showed 99.6% and 99.5 % purity respectively. The IC50 value of the AGY1 against 2D MP-2 was found to be 12.2 ± 1 µM, while the IC50 value of AGY2 was determined to be 2.9 ± 0.2 µM. Both AGY1 and AGY2 anticancer activities were significantly higher than 5-FU (18.5 ± 1.1 µM). However, AGY2 was more effective compared to AGY1. A similar trend was observed for AGY2 analog where the IC50 value for treated 2D PANC-1 was 3.1 ± 0.29 µM and demonstrated a higher anticancer activity compared to AGY1 (6.4 ± 0.38 µM) or 5-FU (7.5 ± 1.1 µM). For cell migration studies, AGY2 (43 ± 5.5) exposed MP-2 cells showed significant arrest towards the wound gap compared to AGY1 (74 ± 18.5) and 5-FU (86 ± 15.6). A similar result of cell migration arrests AGY2 was observed in PANC-1 treated-AGY1, AGY2, and 5-FU. Organoid and spheroid models demonstrated high and comparable anticancer activity of AGY1 and AGY2 compared to 5-FU treated organoid and spheroid models. The cell-cycle analysis showed that AGY2-treated MP-2 cells at 1μΜ concentration showed a higher G1 population arrest (64.25%) compared to AGY1 (54.2%) and 5-FU (55.75%). Similarly, in PANC-1 cells, AGY2 at 1μΜ concentration showed a higher S-phase arrest (53.86%) compared to AGY1 (50.76%) and 5-FU (36.14%). Metabolic stability data showed higher stability of AGY2 (95% intact) compared to AGY1 (85% intact) compared to 5-FU (55% intact) after 2 hour of human liver microsomal incubation. The expressions of p53 and BAX proteins in PANC-1 cells were significantly upregulated and PARP was downregulated at higher concentration when treated with AGY1 and AGY2 compared to 5-FU. Conclusion: We demonstrate that AGY2 showed more promising anticancer activity compared to AGY1 and 5-FU against pancreatic cancer cells. AGY2 has the potential to improve 5-FU’s metabolic stability and enhance its therapeutic efficacy. Citation Format: Raviteja Bulusu, Joy Okoro, Esther Frimpong, Bo Han, Sherise Rogers, Xue Zhu, Edward Agyare. In vitro evaluation of novel pyrimidine nucleosides for the treatment of pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1823.