Cross-linked low molecular weight glycopeptide-mediated gene delivery: relationship between DNA metabolic stability and the level of transient gene expression in vivo.
DNA co-condensates were formed by reacting [125I]DNA with an admixture of a high-mannose glycopeptide (Man9-CWK(18)) and either of two poly(ethylene glycol) peptides (PEG-VS-CWK(18) or PEG-SS-CWK(18)) followed by cross-linking with 6-50 mol equiv of glutaraldehyde. [125I]DNA co-condensates were administered intravenously in mice to determine the influence of peptide DNA formulation parameters on specific targeting to Kupffer cells. Optimal targeting to Kupffer cells required the combined use of 50 mol % Man9-CWK(18) and PEG-CWK(18) to mediate specific recognition by the mannose receptor to Kupffer cells. The cellular uptake of cross-linked Man9-CWK(18)/PEG-CWK(18) DNA co-condensates was receptor mediated since Kupffer cell targeting was inhibited by pre-administration of Man-bovine serum albumin (BSA) but not BSA. An optimized formulation targeted 60% of the dose to the liver, with 80% of the liver-targeted DNA localized to Kupffer cells. Cross-linking with either 6, 15, or 50 mol equiv of glutaraldehyde led to a corresponding decrease in the metabolism rate of DNA in liver as measured by half-live- of 4, 6, and 39 h, respectively. Tail vein dosing of 50 microg of DNA co-condensates cross-linked with 6 mol equiv of glutaraldehyde produced detectable levels of human alpha1-antitrypsin in blood after 12 h, which peaked at day six and persisted for 10 days. The level of human alpha1-antitrypsin was elevated two-fold each day when dosing with DNA co-condensates cross-linked with 15 mol equiv of glutaraldehyde, revealing a correlation between the metabolic stability of the DNA in liver and level of gene expression. In addition to possessing greater metabolic stability, DNA co-condensates cross-linked with 50 mol equiv of glutaraldehyde, but lacking a targeting ligand, avoided rapid liver uptake and possessed a prolonged pharmacokinetic half-life, providing insight into a means to target DNA condensates to peripheral tissues.
Yang
Y
Y
Departments of Medicinal Chemistry and Pharmaceutics, College of Pharmacy, University of Michigan, Ann Arbor, Michigan 48109-1065, USA.
Park
Y
Y
Man
S
S
Liu
Y
Y
Rice
K G
KG
D000818:Animals; D002626:Chemistry, Pharmaceutical; D003432:Cross-Linking Reagents / Q000008:administration & dosage / Q000737:chemistry / Q000493:pharmacokinetics*; D004247:DNA / Q000008:administration & dosage / Q000378:metabolism* / Q000493:pharmacokinetics; D004337:Drug Carriers / Q000737:chemistry / Q000378:metabolism / Q000493:pharmacokinetics*; D005786:Gene Expression Regulation; D006020:Glycopeptides / Q000008:administration & dosage / Q000737:chemistry / Q000493:pharmacokinetics*; D007728:Kupffer Cells / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D051379:Mice; D008813:Mice, Inbred ICR; D008970:Molecular Weight
D003432:Cross-Linking Reagents; D004337:Drug Carriers; D006020:Glycopeptides; D004247:DNA