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Rational Design of Cholesterol Derivative for Improved Stability of Paclitaxel Cationic Liposomes

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Abstract

Purpose

This work explores synthesis of novel cholesterol derivative for the preparation of cationic liposomes and its interaction with Paclitaxel (PTX) within liposome membrane using molecular dynamic (MD) simulation and in-vitro studies.

Methods

Cholesteryl Arginine Ethylester (CAE) was synthesized and characterized. Cationic liposomes were prepared using Soy PC (SPC) at a molar ratio of 77.5:15:7.5 of SPC/CAE/PTX. Conventional liposomes were composed of SPC/cholesterol/PTX (92:5:3 M ratio). The interaction between paclitaxel, ligand and the membrane was studied using 10 ns MD simulation. The interactions were studied using Differential Scanning Calorimetry (DSC) and Small Angle Neutron Scattering analysis. The efficacy of liposomes was evaluated by MTT assay and endothelial cell migration assay on different cell lines. The safety of the ligand was determined using the Comet Assay.

Results

The cationic liposomes improved loading efficiency and stability compared to conventional liposomes. The increased PTX loading could be attributed to the hydrogen bond between CAE and PTX and deeper penetration of PTX in the bilayer. The DSC study suggested that inclusion of CAE in the DPPC bilayer eliminates Tg. SANS data showed that CAE has more pronounced membrane thickening effect as compared to cholesterol. The cationic liposomes showed slightly improved cytotoxicity in three different cell lines and improved endothelial cell migration inhibition compared to conventional liposomes. Furthermore, the COMET assay showed that CAE alone does not show any genotoxicity.

Conclusions

The novel cationic ligand (CAE) retains paclitaxel within the phospholipid bilayer and helps in improved drug loading and physical stability.

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Abbreviations

1HNMR:

Proton nuclear magnetic resonance

CAE:

Cholesteryl arginine ethylester

DMEM:

Dulbecco’s modified eagle’s medium

DPX:

Disterene plasticizer xylene

H5V:

Mouse endothelial cell line

HDMEC:

Human dermal microvascular endothelial cells

IC50:

Concentration at which 50% inhibition seen

IntraHB:

Intramolecular hydrogen bonds

LMP:

Low melting point

MD Simulation:

Molecular dynamic simulation

MDA-MB 231:

Human breast cancer adenocarcinoma cell line

MolSA:

Molecular surface area

MTT:

3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide

OD:

Optical density

OPLS3:

Optimized potentials for liquid simulations

POPC:

1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine

PSA:

Polar surface area

PTX:

Paclitaxel

RMSD:

Relative mean square deviation

SASA:

Solvent accessible surface area

SPC:

Soy phosphatidylcholine

TIP3P:

Transferable intermolecular potential with 3 points

TLC:

Thin layer chromatography

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Authors and Affiliations

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

    Jasmin Monpara & Pradeep R. Vavia

  2. Tumor Microcirculation Group, Department of Oncology & Metabolism School of Medicine, The University of Sheffield, Sheffield, UK

    Chryso Kanthou & Gillian M. Tozer

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  1. Jasmin Monpara
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  2. Chryso Kanthou
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  3. Gillian M. Tozer
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  4. Pradeep R. Vavia
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Correspondence to Pradeep R. Vavia.

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Monpara, J., Kanthou, C., Tozer, G.M. et al. Rational Design of Cholesterol Derivative for Improved Stability of Paclitaxel Cationic Liposomes. Pharm Res 35, 90 (2018). https://doi.org/10.1007/s11095-018-2367-8

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  • DOI: https://doi.org/10.1007/s11095-018-2367-8

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