Abstract 1828: Development and evaluation of novel gemcitabine analog for the treatment of pancreatic cancer

Abstract

Abstract Purpose: Gemcitabine acts against pancreatic cancer and a wide range of solid tumors and it is known to be rapidly deaminated or metabolized in the blood to an inactive metabolite. To increase its therapeutic levels, gemcitabine is administered at high doses causing severe side effects. To improve its metabolic stability, increase cytotoxic activity, and limit the phenomena of resistance many alternatives have emerged, such as modifying gemcitabine. The purpose of this study was to develop and characterize a novel gemcitabine analog with improved stability and enhanced anticancer activity against pancreatic cancer. Methods: A novel analog of gemcitabine (GemAGY) was designed and synthesized through chemical modification of gemcitabine hydrochloride (GemHCl) at position 4 with hydroxylamine. Gem AGY was characterized using nuclear magnetic resonance (NMR), micro-elemental analysis, and purity determined using High-Performance Liquid Chromatography (HPLC). Pancreatic cancer cell lines (MiaPaCa-2, BxPC3, and PANC-1) were treated with GemHCl and GemAGY. Cytotoxicity, cell migration, clonogenic assay, cell cycle, and apoptosis studies were performed to evaluate the effect of GemAGY against pancreatic cancer cell lines. Results: The percent purity of GemAGY was over 99.6%. GemAGY demonstrated a higher cytotoxic effect against pancreatic cancer cells with a lower half-maximal inhibitory concentration (IC50) value (2.5-fold high) compared with GemHCl. For MiaPaCa-2 cells, the IC50 value of GemAGY was found to be 1.63 ± 0.15 µm compared to GemHCl (4.0 ± 1.7 µm). GemAGY also exhibited higher cytotoxicity in PANC-1 (IC50: 1.7 ± 0.2µm) compared to GemHCl (IC50: 5.6 ±1.3 µm) and the IC50 value of GemAGY (3.2±1.1µm) in BxPC-3 cells was significantly better than GemHCl (5.5±1.0 µm). The cell cycle analysis indicated that GemAGY-treated MiaPaCa-2 cells at 1.50 µm concentration had a higher G0/G1 population (60.24%) compared to GemHCl-treated cells (55%). The cell migration studies showed that GemAGY-treated MiaPaCa-2 cells at 12.5 µm concentration significantly inhibited cell mobility towards the wound area with migrated cells found to be 65 ± 6 compared to Gem HCl (174.3 ± 4.0) (GemAGY vs GemHCl; p<0.0001). Similarly, GemAGY-treated PANC-1 cells at 12.5 µm concentration significantly inhibited cell mobility towards the wound area with (98 ± 5.1) compared to Gem HCl (254 ± 6.6). The quantitative analysis of apoptosis studies showed that GemAGY significantly induced higher apoptosis in a dose-dependent manner in MiaPaCa-2 cells compared to GemHCl as concentration increases (GemAGY vs GemHCl; p <0.0001). Conclusion: This study demonstrates that GemAGY increases the anticancer activity of gemcitabine and may have the potential to improve the chemotherapeutic efficacy of gemcitabine in the treatment of pancreatic cancer. Citation Format: Joy Okoro, Raviteja Bulusu, Esther K. Frimpong, Sherise Rodgers, Bo Han, Xue Zhu, Edward Agyare. Development and evaluation of novel gemcitabine analog 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 1828.