Abstract PO-047: Optimizing the efficacy of 5-FU as a chemotherapeutic agent in advanced pancreatic ductal adenocarcinoma (PDAC) using MIAPaCa-2 and PANC-1 cells

Abstract

Abstract Purpose: Pancreatic ductal adenocarcinoma (PDAC) is the most aggressive and scourging soft tissue tumor worldwide. Its current incidence is 13 per 100,000 US population with a very low 5-years survival rate (5%) and estimated to be the second leading cause of cancer-related death by 2030. Most diagnoses are made at advanced disease states warranting combination chemotherapy. Fluorouracil (5-FU) is an effective drug in treating PDAC; however, rapid degradation and systemic instability remain a drawback to its efficacy. The objective of this study was to chemically modify 5-FU to 1,3 bistetrahydrofuran-2yl-5-FU (MFU) to prolong its systemic stability and enhance the anticancer activity of 5-FU. Method: 5-FU was chemically modified to MFU using tetrahydrofuran-2-yl acetate and 1,8-Diazabicyclo (5.4. 0)undec-7-ene (DBU) in Dimethyl Formamide (DMF). MFU was characterized using nuclear magnetic resonance (NMR) to determine new bond formation, mass spectrometer (MS) for molecular weight determination, high-performance liquid chromatography (HPLC) to determine percent purity, and micro-elemental analysis used to ascertain the presence of elemental composition and purity. Percent conversion of MFU to 5FU was determined using PDAC cells. In vitro anticancer activity of MFU was tested using 2D and 3D MiaPaCa-2 and PANC-1 PDAC cultures and cell viability study performed using the alamar blue assay. Apoptotic and cell cycle studies were performed on MFU treated MiaPaCa-2 and PANC-1 cells using flow cytometry. Results: Molecular weight of synthesized MFU (C12H15FN2O4) was determined to be 270.02 using the Electrospray Ionization (ESI) (MS (M+H)+ = 270.08). While percent purity of MFU was found to be greater than 99.6%, melting point was determined to be 115±2 °C and retention time (RT) was 1.85 minutes. For MFU conversion to 5-FU studies, 4.1% of 25µM and 7.6% of 50µM of MFU incubated MiaPaCa-2 cells was converted to 5-FU over a period of 24 hr showing an incremental production of 5-FU from MFU based on concentration. For in vitro study, half-maximal inhibitory concentration (IC50) of 5-FU treated 2D MiaPaCa-2 culture was 3.4 ±1.1µM while that of MFU treated 2D MiaPaCa-2 culture was 2.1± 0.2 µM. The IC50 value of 5-FU treated 3D MiaPaCa-2 culture was found to be (8.5±1.2µM), while IC50 value for MFU treated 3D MiaPaCa-2 culture was (7.2±1.1µM). Put together, IC50 value for MFU for either 2D or 3D was significantly lower compared with IC50 value 5-FU (P=0.021 (MFU vs 5-FU) for 2D and P=0.04 (MFU vs 5-FU) for 3D). A similar trend was noted when PANC-1 cells were used. Apoptotic and cell cycle studies were similar to that of the 5-FU with most of the cells (54%) in late apoptosis after 48 h of treatment and 64% vs 33% of cells arrested at the G1 and S phase of the cell cycle respectively. Conclusion: The studies demonstrated that MFU may be an alternate approach to improve the delivery, systemic stability, and efficacy of 5-FU in the treatment of PDAC tumors. Citation Format: Nkafu Bechem Ndemazie, Andriana Inkoom, Xue Y. Zhu, Edward Agyare. Optimizing the efficacy of 5-FU as a chemotherapeutic agent in advanced pancreatic ductal adenocarcinoma (PDAC) using MIAPaCa-2 and PANC-1 cells [abstract]. In: Proceedings of the AACR Virtual Special Conference on Pancreatic Cancer; 2021 Sep 29-30. Philadelphia (PA): AACR; Cancer Res 2021;81(22 Suppl):Abstract nr PO-047.