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Cationic cholesterol derivative efficiently delivers the genes: in silico and in vitro studies

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Abstract

The aims of the research work were to synthesize ethyl(cholesteryl carbamoyl)-l-arginate (ECCA), an arginine-conjugated cholesterol derivative, and to evaluate its application as a gene delivery vector. The interactions of ECCA with DNA duplex were studied using molecular dynamics (MD) simulations. It was found that the guanidine group of ECCA could interact with the phosphate group of DNA through ionic interactions as well as hydrogen bonds. The structure of DNA was stable throughout the simulation time. Liposomes were formulated using ECCA and soya phosphatidylcholine (SPC) by a thin-film hydration method. They had the particle size of ~ 150 nm and the zeta potential of + 51 mV. To ensure the efficient binding of DNA to the liposomes, the ratio of DNA to ECCA was optimized using gel retardation assay. Further, serum stability, haemolysis and cytotoxicity studies were carried out to determine the stability and safety of the lipoplexes. Circular dichroism spectroscopy was used to determine the interaction of DNA and cationic liposomes. Cellular uptake pathway was determined by studying the uptake of coumarin-loaded lipoplexes at 4 °C and in the presence of uptake inhibitors, i.e. genistein, chlorpromazine and methyl-β-cyclodextrin. Transfection studies were carried out to evaluate the transfection efficacy of the ECCA-loaded lipoplexes. The binding of DNA and lipoplexes was found to be stable in the presence of serum, and no degradation of DNA was observed. The lipoplexes showed low haemolysis and cytotoxicity. The uptake of coumarin-loaded liposomes was decreased up to ~ 20% in the presence of clathrin- and caveola-mediated uptake inhibitors, indicating a role of both the pathways in the uptake of the inhibitors. Satisfactory transfection efficiency was obtained compared to Lipofectamine®. Thus, cationic cholesterol derivative is a useful tool for gene delivery vector.

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Acknowledgments

Authors are extremely thankful to Dr. Dhanashree Jagtap, National Institute for Research in Reproductive Health (NIRRH), India, for helping with the CD spectrometry study. Ms. Tripti Verma contributed to the manuscript by carrying out the serum stability study.

Funding

The authors are thankful to the University Grant Commission (UGC), Government of India, for the financial assistance and AICTE-NAFETIC for providing facilities to perform the experimental work.

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences and Technology, Institute of Chemical Technology, University Under Section 3 of UGC Act—1956, Elite Status and Center of Excellence—Government of Maharashtra, TEQIP Phase II Funded, Mumbai, 400019, India

    Jasmin Monpara & Pradeep Vavia

  2. Gupta Laboratory, Epigenetics and Chromatin Biology Group, Cancer Research Institute, Tata Memorial Centre, Advanced Center for Treatment, Research and Education in Cancer (ACTREC), Navi Mumbai, 410210, India

    Divya Velga, Tripti Verma & Sanjay Gupta

  3. Homi Bhabha National Institute, Training School Complex, Anushakti Nagar, Mumbai, MH, 400085, India

    Divya Velga, Tripti Verma & Sanjay Gupta

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  1. Jasmin Monpara
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Correspondence to Pradeep Vavia.

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Monpara, J., Velga, D., Verma, T. et al. Cationic cholesterol derivative efficiently delivers the genes: in silico and in vitro studies. Drug Deliv. and Transl. Res. 9, 106–122 (2019). https://doi.org/10.1007/s13346-018-0571-z

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