Codelivery of doxorubicin and JIP1 siRNA with novel EphA2-targeted PEGylated cationic nanoliposomes to overcome osteosarcoma multidrug resistance

Fateme Haghiralsadat,1–3 Ghasem Amoabediny,3–5 Samira Naderinezhad,4 Behrouz Zandieh-Doulabi,6 Tymour Forouzanfar,5 Marco N Helder5 1Department of Life Science Engineering, Faculty of New Sciences and Technologies, University of Tehran, Tehran, Iran; 2Department of Orthopaedic Surgery, VU University Medical Center, MOVE Research Institute Amsterdam, Amsterdam, the Netherlands; 3Department of Nano Biotechnology, Research Center for New Technologies in Life Science Engineering, 4Department of Biotechnology and Pharmaceutical Engineering, School of Chemical Engineering, College of Engineering, University of Tehran, Tehran, Iran; 5Department of Oral and Maxillofacial Surgery, VU University Medical Center, MOVE Research Institute Amsterdam, 6Department of Oral Cell Biology and Functional Anatomy, Academic Center for Dentistry Amsterdam (ACTA), Vrije Universiteit Amsterdam and University of Amsterdam, MOVE Research Institute, Amsterdam, the Netherlands Purpose: Osteosarcoma (OS) mostly affects children and young adults, and has only a 20%–30% 5-year survival rate when metastasized. We aimed to create dual-targeted (extracellular against EphA2 and intracellular against JNK-interacting protein 1 [JIP1]), doxorubicin (DOX)-loaded liposomes to treat OS metastatic disease. Materials and methods: Cationic liposomes contained N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium methyl-sulfate (DOTAP), cholesterol, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), and distearoyl-phosphatidylethanolamine–methyl-poly(ethylene glycol) (DSPE–mPEG) conjugate. EphA2 targeting was accomplished by conjugating YSA peptide to DSPE–mPEG. Vesicles were subsequently loaded with DOX and JIP1 siRNA. Results: Characteristics assessment showed that 1) size of the bilayered particles was 109 nm; 2) DOX loading efficiency was 87%; 3) siRNA could be successfully loaded at a liposome:siRNA ratio of >24:1; and 4) the zeta potential was 18.47 mV. Tumor-mimicking pH conditions exhibited 80% siRNA and 50.7% DOX sustained release from the particles. Stability studies ensured the protection of siRNA against degradation in serum. OS cell lines showed increased and more pericellular/nuclear localizations when using targeted vesicles. Nontargeted and targeted codelivery caused 70.5% and 78.6% cytotoxicity in OS cells, respectively (free DOX: 50%). Targeted codelivery resulted in 42% reduction in the siRNA target, JIP1 mRNA, and 46% decrease in JIP1 levels. Conclusion: Our dual-targeted, DOX-loaded liposomes enhance toxicity toward OS cells and may be effective for the treatment of metastatic OS. Keywords: MAP kinase 8 interacting protein 1, MAPK8IP1, functionalization, cationic liposome, intracellular targeting, extracellular targetin

A comprehensive mathematical model of drug release kinetics from nano-liposomes, derived from optimization studies of cationic PEGylated liposomal doxorubicin formulations for drug-gene delivery

This study focuses on the development of a universal mathematical model for drug release kinetics from liposomes to allow in silico prediction of optimal conditions for fine-tuned controlled drug release. As a prelude for combined siRNA-drug delivery, nanoliposome formulations were optimized using various mole percentages of a cationic lipid (1,2-dioleoyl-3-trimethylammonium-propane, DOTAP) in the presence or absence of 3 mol% distearoyl phosphoethanolamine, polyethylene glycol (PEG–2000mDSPE). Outcome parameters were particle size, zeta potential, entrapment efficiency, in vitro drug release, and tumor cell kill efficiency. The optimized formula (containing 20% DOTAP with 3% DSPE-mPEG(2000) was found to be stable for six months, with round-shaped particles without aggregate formation, an average diameter of 71 nm, a suitable positive charge, and 89% drug encapsulation efficiency (EE). The 41% drug release during 6 h confirmed controlled release. Furthermore, the release profiles as functions of pH and temperature were investigated and the kinetics of the drug release could adequately be fitted to Korsmeyer–Peppas’ model by multiple regression analysis. The statistical parameters confirmed good conformity of final models. Functionality of the novel cationic liposome formulations (± DOX) was tested on osteosarcoma (OS) cell lines. Increased OS cell toxicity (1.3-fold) was observed by the DOX-loaded vs. the free DOX. A feasibility pilot showed that siRNA could be loaded efficiently as well. In conclusion, we have established a predictive mathematical model for the fine-tuning of controlled drug release from liposomal formulations, while creating functional drug-delivery liposomes with potential for siRNA co-delivery to increase specificity and efficacy. (Figure presented.

New liposomal doxorubicin nanoformulation for osteosarcoma:Drug release kinetic study based on thermo and pH sensitivity

A novel approach was developed for the preparation of stealth controlled-release liposomal doxorubicin. Various liposomal formulations were prepared by employing both thin film and pH gradient hydration techniques. The optimum formulation contained phospholipid and cholesterol in 1:0.43 molar ratios in the presence of 3% DSPE-mPEG (2000). The liposomal formulation was evaluated by determining mean size of vesicle, encapsulation efficiency, polydispersity index, zeta potentials, carrier's functionalization, and surface morphology. The vesicle size, encapsulation efficiency, polydispersity index, and zeta potentials of purposed formula were 93.61 nm, 82.8%, 0.14, and −23, respectively. Vesicles were round-shaped and smooth-surfaced entities with sharp boundaries. In addition, two colorimetric methods for cytotoxicity assay were compared and the IC50 (the half maximal inhibitory concentration) of both methods for encapsulated doxorubicin was determined to be 0.1 μg/ml. The results of kinetic drug release were investigated at several different temperatures and pH levels, which showed that purposed formulation was thermo and pH sensitive

New Approach for the Synthesis, Entrapment of Hydrophilic Drugs and Evaluation of Physico-Chemical Characteristics of PH-sensitive Nano-Liposome: Improving Therapeutic Efficacy of Doxorubicin in Order to Treatment Bone Tumor and Reducing the Side Effects of Doxorubicin

Introduction: Cancer is one of the most harmful disease throughout the world. Doxorubicin is an anti-cancer agent, used in the treatment of various types of the cancer such as bone cancer. There are several adverse effects related to clinical usage of Doxorubicin for long time. The present study aimed to investigate the reducing side effects and enhancing the therapeutic effect by liposomal carrier. Methods: Liposomes containing DPPG and cholesterol with the molar ratio of 70:30 with the Doxorubicin were synthesized by pH- gradient method. The average diameter of nanoparticles and surface charge was determined by Zeta-Sizer instrument. The amount of drug loaded and drug-released was determined using dialysis. The surface morphology and internal lamella was evaluated by TEM and SEM. Results: The average size of liposomal Doxorubicin obtained using Zeta-Sizer was 126 nm. The encapsulation efficacy of liposomal Doxorubicin was 89%. The total amount of drug release during 48 hours in acidic medium studied by dialysis technique was 46%. Conclusion: In this study, investigation of loading Doxorubicin into nano-liposome with the slow- released kinetic was carried out to improve the solubility and bioavailability of Doxorubicin in order to delivery to osteosarcoma cell line