Abstract 4477: Biological evaluation of EGFR- conjugated liposomal drug delivery system for enhanced targeting of pancreatic cancer

Abstract Purpose: The purpose of this study is to entrap and characterize a potential lead compound (heptyl (5-fluoro-2-oxo-1-(tetrahydrofuran-2-yl)-1, 2-dihydropyrimidine-4-yl) carbamate (AGY2) into antibody-conjugated liposomal drug delivery system (AGY2-EGFR LDDS) to target pancreatic cancer. Methods: The monoclonal antibody (Cetuximab) conjugated AGY2 loaded liposome drug delivery system (AGY2-LDDS) using DPPC, MPPC, and PEG-2000 lipids. The conjugation was confirmed by Fourier Transform Infrared Spectroscopy (FTIR) spectra. The particle size (PS), zeta potential (ZP), polydispersity index (PI), and entrapment efficiency (EE) of the formulation were determined. 3D cytotoxicity studies were performed on MiaPaCa-2 (MP-2) and PANC-1 (P-1) cells. Stability study was performed at different temperatures (4°C, 25°C, and 40°C). Pharmacokinetic (PK) and biodistribution studies were conducted. A tumor efficacy study was conducted by inoculating the KPCY PDX mice. Results: The variations in the peaks for AGY2, AGY2 LDDS, and AGY2-EGFR LDDS in the FTIR spectra confirmed the successful conjugation of EGFR to the lipids. The PS of the formulation was 115±7 nm with a ZP of -35 ±8 mV, and the EE was 80%, as analyzed by Stat-Ease360. In MP-2 spheroids, the IC50 values were as follows: AGY2-EGFR LDDS at 7.7 ± 2.3 µM (p < 0.0001), AGY2 LDDS at 16.5 ± 3.7 µM (p < 0.001), and AGY2 at 27.5 ± 4.2 µM (p < 0.0328), compared to 5-Fluorouracil (5-FU) at 36.6 ± 4.8 µM. Similarly, in P-1 cells, the IC50 values were: AGY2-EGFR LDDS at 9.6 ± 0.9 µM (p < 0.0001), AGY2 LDDS at 18.5 ± 5.7 µM (p < 0.0003), and AGY2 at 35.4 ± 5.2 µM, compared to 5-FU at 40.8 ± 5.9 µM. In MP-2 organoids, the IC50 values were: AGY2-EGFR LDDS at 10.8 ± 3.3 µM (p < 0.0001), AGY2 LDDS at 25.1 ± 3.9 µM (p < 0.0004), and AGY2 at 35.2 ± 3.7 µM (p < 0.0154), compared to 5-FU at 47.9 ± 4.4 µM. Similarly, in P-1 organoids, the IC50 values were: AGY2-EGFR LDDS at 12.2 ± 3.9 µM (p < 0.0001), AGY2 LDDS at 29.09 ± 4.9 µM (p < 0.001), and AGY2 at 41.56 ± 5.1 µM, compared to 5-FU at 45.96 ± 4.8 µM. Stability studies showed that at 4°C and 25°C, the formulations remained stable over 30, 60, and 90 days, with no significant changes in PS and EE. However, at 40°C, the formulations were stable for 30 and 60 days, but the 90-day samples showed reduced EE and increased PS. The PK study demonstrated that AGY2-EGFR LDDS had a threefold increase in half-life (1.54±0.06 vs. 0.49±0.035-FU h), AUC (428±10 vs. 125±125-FU µg/L*h), and clearance (0.73 vs. 2.55-FU L/h). The biodistribution study showed a higher AGY2-EGFR LDDS tumor accumulation compared to 5-FU. In the tumor efficacy study, the AGY2-EGFR LDDS treated group showed significant tumor growth inhibition compared to the 5-FU treated group. Conclusion: This study shows that AGY2-EGFR LDDS improves the anticancer activity 5-FU and may have the potential to significantly reduce tumor growth in EGFR-positive pancreatic cancer. Citation Format: Raviteja Bulusu, Joy Okoro, Esther Frimpong, Saunjoo Yoon, Bo Han, Renee Reams, Zhu Xue, Edward Agyare. Biological evaluation of EGFR- conjugated liposomal drug delivery system for enhanced targeting of pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 4477.