RESUMO
Although adenovirus is a popular vector for delivering genes, there are several drawbacks that limit its effectiveness, including tropism and both the innate and adaptive immune responses. One approach that has been used to ameliorate these drawbacks is PEGylation of the virus with subsequent modification to add functional moieties for the purpose of cell targeting or enhancing infection. Here, we describe a general approach for PEGylating adenovirus and conjugating cell-penetrating peptides to the surface of the virus to impart the ability to transduce CAR-negative cells.
Assuntos
Adenoviridae/imunologia , Proteínas do Capsídeo/imunologia , Vetores Genéticos/efeitos adversos , Interações Hospedeiro-Patógeno/imunologia , Adenoviridae/genética , Sequência de Aminoácidos , Animais , Antígenos Virais/química , Antígenos Virais/imunologia , Proteínas do Capsídeo/química , Proteínas do Capsídeo/genética , Peptídeos Penetradores de Células/química , Técnicas de Transferência de Genes , Terapia Genética/efeitos adversos , Vetores Genéticos/genética , Camundongos , Estrutura Molecular , Células NIH 3T3 , Polietilenoglicóis/química , Transdução GenéticaRESUMO
The aim of this study was to investigate the influence of the nanocarrier surface charge on brain delivery of a model hydrophilic drug via the nasal route. Anionic and cationic nanostructured lipid carriers (NLCs) were prepared and optimized for their particle size and zeta potential. The optimum particles were incorporated in poloxamer in situ gels and their in vivo behavior was studied in the plasma and brain after administration to rats. Optimum anionic and cationic NLCs of size <200 nm and absolute zeta potential value of ≈ 34 mV were obtained. Toxicity study revealed mild to moderate reversible inflammation of the nasal epithelium in rats treated with the anionic NLCs (A7), and destruction of the lining mucosal nasal epithelium in rats treated with the cationic NLCs (C7L). The absolute bioavailability of both drug loaded anionic and cationic NLCs in situ gels was enhanced compared to that of the intranasal solution (IN) of the drug with values of 44% and 77.3%, respectively. Cationic NLCs in situ gel showed a non significant higher Cmax (maximum concentration) in the brain compared to the anionic NLCs in situ gel. Anionic NLCs in situ gel gave highest drug targeting efficiency in the brain (DTE%) with a value of 158.5 which is nearly 1.2 times that of the cationic NLCs in situ gel.