Rational Design of Cholesterol Derivative for Improved Stability of Paclitaxel Cationic Liposomes
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.
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.
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.
The novel cationic ligand (CAE) retains paclitaxel within the phospholipid bilayer and helps in improved drug loading and physical stability.
KEY WORDSCOMET assay molecular dynamic simulation paclitaxel-loaded cationic liposomes trans-well migration assay
Proton nuclear magnetic resonance
Cholesteryl arginine ethylester
Dulbecco’s modified eagle’s medium
Disterene plasticizer xylene
Mouse endothelial cell line
Human dermal microvascular endothelial cells
Concentration at which 50% inhibition seen
Intramolecular hydrogen bonds
Low melting point
- MD Simulation
Molecular dynamic simulation
- MDA-MB 231
Human breast cancer adenocarcinoma cell line
Molecular surface area
Optimized potentials for liquid simulations
Polar surface area
Relative mean square deviation
Solvent accessible surface area
Transferable intermolecular potential with 3 points
Thin layer chromatography
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