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Liposome-encapsulated EF24-HPβCD inclusion complex: a preformulation study and biodistribution in a rat model

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Abstract

3,5-Bis(2-fluorobenzylidene)-4-piperidone (EF24) is an anti-proliferative diphenyldifluoroketone analog of curcumin with more potent activity. The authors describe a liposome preparation of EF24 using a “drug-in-CD-in liposome” approach. An aqueous solution of EF24 and hydroxypropyl-β-cyclodextrin (HPβCD) inclusion complex (IC) was used to prepare EF24 liposomes. The liposome size was reduced by a combination of multiple freeze–thaw cycles. Co-encapsulation of glutathione inside the liposomes conferred them with the capability of labeling with imageable radionuclide Tc-99m. Phase solubility analysis of EF24-HPβCD mixture provided k 1:1 value of 9.9 M−1. The enhanced aqueous solubility of EF24 (from 1.64 to 13.8 mg/mL) due to the presence of HPβCD helped in the liposome preparation. About 19% of the EF24 IC was encapsulated inside the liposomes (320.5 ± 2.6 nm) by dehydration–rehydration technique. With extrusion technique, the size of 177 ± 6.5 nm was obtained without any effect on encapsulation efficiency. The EF24-liposomes were evaluated for anti-proliferative activity in lung adenocarcinoma H441 and prostate cancer PC-3 cells. The EF24-liposomes demonstrated anti-proliferative activity superior to that of plain EF24 at 10 μM dose. When injected in rats, the Tc-99m-labeled EF24-liposomes cleared from blood with an α-t 1/2 of 21.4 min and β-t 1/2 of 397 min. Tissue radioactivity counting upon necropsy showed that the majority of clearance was due to the uptake in liver and spleen. The results suggest that using “drug-in-CD-in liposome” approach is a feasible strategy to formulate an effective parenteral preparation of EF24. In vitro studies show that the liposomal EF24 remains anti-proliferative, while presenting an opportunity to image its biodistribution.

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Acknowledgments

The authors acknowledge the help of Ms. Prachi Vilekar (Graduate Student) in cell culture studies. The radiolabeling and biodistribution study was performed in the Small Animal Imaging Facility of the College of Pharmacy. This study was made possible by the financial support under the National Cancer Institute grant 1R03 CA143614-01.

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, University of Oklahoma Health Sciences Center, 1110 N. Stonewall Avenue, Oklahoma City, OK, 73117, USA

    H. Agashe, P. Lagisetty, K. Sahoo, D. Bourne & V. Awasthi

  2. School of Chemical, Biological and Materials Engineering, 100 East Boyd, Norman, OK, 73019, USA

    B. Grady

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  1. H. Agashe
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  2. P. Lagisetty
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Correspondence to V. Awasthi.

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Agashe, H., Lagisetty, P., Sahoo, K. et al. Liposome-encapsulated EF24-HPβCD inclusion complex: a preformulation study and biodistribution in a rat model. J Nanopart Res 13, 2609–2623 (2011). https://doi.org/10.1007/s11051-010-0154-5

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