Tissue Distribution of the Cytofectin Component of a Plasmid–DNA/Cationic Lipid Complex Following Intravenous Administration in Mice

Abstract

Allovectin-7 is a gene therapy agent that consists of plasmid DNA (pDNA) encoding the human HLA-B7 class I and β ₂-microglobulin genes (VCL-1005), complexed with the cationic lipid DMRIE Br and DOPE. A tritiated version of the cytofectin component, DMRIE Br, was synthesized by regiospecific isotope incorporation to a very high specific activity. The ³H-labeled DMRIE/DOPE mixture was complexed with VCL-1005 to produce a radiolabeled version of Allovectin-7. The VCL-1005/³H-DMRIE/DOPE complex was administered intravenously to mice, and the tissue distribution of radioactivity was analyzed 24 hr later. Excretion of radioisotope was monitored for 96 hr post dosing. At 24 hr post administration, a tissue distribution for the radioisotope of liver ≫ spleen > lung ≫ heart > brain ≈ muscle ≈ blood was observed. During the 96-hr period post dose, very little administered radioactivity (<17%) was excreted and the majority of the isotope (83%) remained in the animal. This is the first report on the biodistribution of the cytofectin component of a pDNA–cationic lipid complex for which the distribution of the plasmid component has also been reported.

Overview summary

The first detailed biodistribution analysis of the cytofectin component of a plasmid DNA–lipid complex is described. Multiple tritium atoms were regiospecifically incorporated into the cytofectin DMRIE Br to afford a very high specific activity material. The tritiated plasmid DNA–lipid complex, VCL-1005/³H-DMRIE/DOPE was prepared and administered intravenously to mice. The radioisotope distribution in tissues was analyzed at 24 hr post dose and the excretion of radioactivity was monitored for 96 hr after administration.