Abstract 839: Characterization and evaluation of modified 5-fluorouracil loaded liposomal nanoparticle on pancreatic and colorectal cancer cell lines
Authors: Bulusu Raviteja , Ndemazie Nkafu B. , Frimpong Esther K. , Inkoom Andriana , Okoro Joy C. , Zhu Xue Y. 
Published: April 2023

The purpose of the study is to characterize modified 5-FU (MFU) and investigate its anticancer activity in liposomal nanoparticles (MFU-Lnps) against colorectal and pancreatic cancer cell lines.

Methods: The synthesized MFU was characterized by NMR and elemental analysis and formulated as MFU-Lnps. The particle size, polydispersity index (PDI), and zeta potential were determined using a particle size analyzer. The in-vitro studies were performed on 2D and 3D models of HCT-116 and MiaPaCa-2 cell lines. Cellular uptake of rhodamine (Rho) (cationic dye) alone or Rho-linked-Lnps (Rho-Lnp) was determined by confocal fluorescence microscopy. In-vitro release of MFU from MFU-Lnps was performed using PBS and methanol (3:1) at 37° C under sink conditions. To evaluate the pharmacokinetics (PK) parameters and tissue biodistribution behavior of Lnp, Lnp was loaded with gadolinium-hexanoate (Gad-Hex-Lnps), and the quantified by inductively coupled plasma mass spectrometry (ICP-MS).

Results: The 1H NMR peaks at 1.02 ppm (-C-N on 2') and 1.04 ppm (-C-N on 2''), represent the amine bond between 4-NH2 group of 5-FU and tetrahydrofuran acetate. This confirmed that the MFU was successfully synthesized. The elemental analysis of MFU showed 99.5% purity by comparing observed and theoretical values (Observed: carbon (C) = 53.57 %, hydrogen (H)= 5.64 %, and nitrogen (N) =10.44 % and, theoretical: C=53.33%, H=5.59%, and N=10.37 %). The MFU-Lnps hydrodynamic diameter was 124.9 ± 3.2 nm, polydispersity was found to be 0.16 ± 0.005, while entrapment efficiency and zeta potential values were 97.2% and -30.3 ± 12 mV, respectively. The MFU treated-2D and 3D MiaPaCa-2 cultures (IC50 values: 3.9 ± 0.9 µM2D and 6.7±1.1µM3D) was more effective than 5-FU-treated 2D and 3D MiaPaCa-2 cultures (IC50 values: 5.4 ± 0.8 µM2D and 8.5 ± 1.0 µM3D). However, MFU-Lnps treated-MiaPaCa-2 3D-organoid was 4-fold more effective compared with MFU (IC50 values: 9.8 ± 1.4 µMMFU-Lnps versus 42.3 ± 2.9 µMMFU). A similar high cytotoxicity effect of MFU-Lnp was observed when MFU-Lnp (IC50 values: 2.0 ± 0.7 µM2D and 3.4 ±0.8 µM3D) was exposed to HCT-116 2D and 3D cultures compared to MFU (3.1± 0.8µM2D and 6.8 ± 0.9 µM3D). For cellular uptake, we observed an increase in the uptake of Rho-linked-Lnps compared with Rho alone. The MFU release MFU-Lnps exhibited Higuchi release kinetics model based on the coefficient of determination (R2). For PK results, the half-life of Gad-Hex-Lnps (32.5 ± 2.6 min) was significantly high compared to that of Gad-Hex (21.5 ± 0.7 min) and, AUC of Gad-Hex-Lnps (1641 ± 499 µg/L*min) was higher than Gad-Hex (932 ± 256 µg/L*min). The accumulation of Gad-Hex-Lnps in the pancreas was remarkably higher than Gad-Hex.

In conclusion, we have demonstrated that MFU-Lnps improves the anticancer efficacy of 5-FU and Lnps delivery system may improve PK parameters and increases the biodistribution of 5-FU.

Citation Format: Raviteja Bulusu, Nkafu B. Ndemazie, Esther K. Frimpong, Andriana Inkoom, Joy C. Okoro, Xue Y. Zhu. Characterization and evaluation of modified 5-fluorouracil loaded liposomal nanoparticle on pancreatic and colorectal cancer cell lines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 839.