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Development and In Vitro Evaluation of Long Circulating Liposomes for Targeted Delivery of Gemcitabine and Irinotecan in Pancreatic Ductal Adenocarcinoma

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Abstract

The classically used nontargeted chemotherapeutic approach to pancreatic cancer has a dual drawback of suboptimal drug delivery at the target site and the systemic side effects produced by the unfettered exposure of the drug to healthy tissue. This study has the objective of developing novel poly(2-ethyl-2-oxazoline) (PETOX)–based long circulating liposomes loaded with gemcitabine and irinotecan for the treatment of pancreatic ductal adenocarcinoma, with a juxtaposition to PEGylated and uncoated liposomes. A PETOX−cholesteryl chloroformate lipopolymer conjugate (PETOX–ChC) with a carbonate linkage was prepared and characterized by 1H NMR, FTIR, and DSC. Liposomes were prepared using the thin film hydration technique followed by freeze–thaw and membrane extrusion methods. Liposome characterization includes particle size determination, zeta potential determination using a zetameter, and structural elucidation using 31P NMR and cryo-TEM. The PETOXylated liposomes showed a particle size of 180.1 ± 2.2 nm and a zeta potential of − 33.63 ± 1.23 mV. The liposomal combination therapy of gemcitabine and irinotecan was found to have an IC50 value 39 times lower in comparison to the drug combination in solution, while the PEGylated and PETOXylated liposomes showed IC50 values 1.6 times lower and 2 times lower than that of uncoated liposomes, respectively, against Mia PaCa II pancreatic cancer cell line. The PEGylated and PETOXylated liposomes showed 4.1 and 5.4 times slower macrophagial uptake in vitro in comparison to the uncoated liposomes respectively. The PEGylated liposomes showed 11% higher in vitro macrophagial uptake in comparison to PETOXylated liposomes.

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Abbreviations

ABC:

Accelerated blood clearance

ChC:

Cholesteryl chloroformate

Cryo-TEM:

Transmission electron cryomicroscopy

DCC:

N,N′-Dicyclohexylcarbodiimide

DCM:

Dichloromethane

DMAP:

4-Dimethylaminopyridine

DSC:

Differential scanning calorimetry

DSPE:

1,2-Distearoyl-sn-glycero-3-phosphoethanolamine

FTIR:

Fourier transform infrared spectroscopy

HPLC:

High-performance liquid chromatography

MTT:

(3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide)

NMR:

Nuclear magnetic resonance

PDAC:

Pancreatic ductal adenocarcinoma

PE:

Phosphatidyl ethanolamine

PEG:

Polyethylene glycol

PETOX:

Poly(2-ethyl-2-oxazoline)

RES:

Reticuloendothelial system

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Acknowledgments

The cryo-TEM images were obtained with cooperation from Dr. Wei Zhang at the University of Minnesota, MN, USA. The pancreatic cancer cell lines were obtained from Dr. Surinder Batra and Dr. Rakesh Singh of the University of Nebraska Medical Center, Omaha, NE.

Funding

This work was supported by the Department of Pharmacy Sciences and the Department of Chemistry at Creighton University, Omaha, NE, USA. The confocal imaging research was conducted at the Integrated Biomedical Imaging Facility at Creighton University, Omaha, NE. This facility is supported by the Creighton University School of Medicine and grants GM103427 and GM110768 from the National Institute of General Medical Science (NIGMS), a component of the National Institutes of Health (NIH). The facility was constructed with support from grants from the National Center for Research Resources (RROI 64 69) and the NIGMS (GM103427).

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Authors and Affiliations

  1. Department of Pharmacy Sciences, School of Pharmacy and Health Professions, Creighton University, 2500 California Plaza, Omaha, Nebraska, 68178, USA

    S. Deodhar, A. K. Dash & E. J. North

  2. Department of Chemistry, Creighton University, Omaha, Nebraska, USA

    M. Hulce

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  1. S. Deodhar
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Correspondence to A. K. Dash.

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Deodhar, S., Dash, A.K., North, E.J. et al. Development and In Vitro Evaluation of Long Circulating Liposomes for Targeted Delivery of Gemcitabine and Irinotecan in Pancreatic Ductal Adenocarcinoma. AAPS PharmSciTech 21, 231 (2020). https://doi.org/10.1208/s12249-020-01745-6

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