An in situ gelling liquid crystalline system based on monoglycerides and polyethylenimine for local delivery of siRNAs

• Published in: European journal of pharmaceutical sciences 2015-07, Vol.74, p.103-117
• Main Authors: Borgheti-Cardoso, Lívia Neves, Depieri, Lívia Vieira, Kooijmans, Sander A.A., Diniz, Henrique, Calzzani, Ricardo Alexandre Junqueira, Vicentini, Fabiana Testa Moura de Carvalho, van der Meel, Roy, Fantini, Márcia Carvalho de Abreu, Iyomasa, Mamie Mizusaki, Schiffelers, Raymond M., Bentley, Maria Vitória Lopes Badra
• Format: Article
• Language: English
• Subjects: Animals; Cellulitis - chemically induced; Cellulitis - immunology; Cellulitis - pathology; Cellulitis - prevention & control; Female; Gels; Gene Transfer Techniques - adverse effects; Glycerides - adverse effects; Glycerides - chemistry; Injections, Subcutaneous; Liquid crystal; Local release; Mice, Inbred BALB C; Monoglycerides - adverse effects; Monoglycerides - chemistry; Non-viral gene delivery; Polyethyleneimine - adverse effects; Polyethyleneimine - chemistry; Propylene Glycol - adverse effects; Propylene Glycol - chemistry; RNA Interference; RNA, Small Interfering - administration & dosage; RNA, Small Interfering - adverse effects; RNA, Small Interfering - chemistry; RNAi Therapeutics - adverse effects; Self-assembly; siRNA; Skin - drug effects; Skin - immunology; Skin - pathology; Solubility; Subcutaneous Tissue - drug effects; Subcutaneous Tissue - immunology; Subcutaneous Tissue - pathology; Viscoelastic Substances - adverse effects; Viscoelastic Substances - chemistry; Viscosity; Water - analysis
• ISSN: 0928-0987; 1879-0720
• DOI: 10.1016/j.ejps.2015.04.017

[Display omitted] The development of delivery systems able to complex and release siRNA into the cytosol is essential for therapeutic use of siRNA. Among the delivery systems, local delivery has advantages over systemic administration. In this study, we developed and characterized non-viral carriers to deliver siRNA locally, based on polyethylenimine (PEI) as gene carrier, and a self-assembling drug delivery system that forms a gel in situ. Liquid crystalline formulations composed of monoglycerides (MO), PEI, propylene glycol (PG) and 0.1M Tris buffer pH 6.5 were developed and characterized by polarized light microscopy, Small Angle X-ray Scattering (SAXS), for their ability to form inverted type liquid crystalline phases (LC2) in contact with excess water, water absorption capacity, ability to complex with siRNA and siRNA release. In addition, gel formation in vivo was determined by subcutaneous injection of the formulations in mice. In water excess, precursor fluid formulations rapidly transformed into a viscous liquid crystalline phase. The presence of PEI influences the liquid crystalline structure of the LC2 formed and was crucial for complexing siRNA. The siRNA was released from the crystalline phase complexed with PEI. The release rate was dependent on the rate of water uptake. The formulation containing MO/PEI/PG/Tris buffer at 7.85:0.65:76.5:15 (w/w/w/w) complexed with 10μM of siRNA, characterized as a mixture of cubic phase (diamond-type) and inverted hexagonal phase (after contact with excess water), showed sustained release for 7days in vitro. In mice, in situ gel formation occurred after subcutaneous injection of the formulations, and the gels were degraded in 30days. Initially a mild inflammatory process occurred in the tissue surrounding the gel; but after 14days the tissue appeared normal. Taken together, this work demonstrates the rational development of an in situ gelling formulation for local release of siRNA.