ABSTRACT
Improved non-viral vector systems are needed for efficient delivery of DNA to target cell nuclei in gene therapy. A series of linear polyamine poly(ethylene glycol) (PEG) constructs has been synthesised by reaction of appropriately Boc-protected thermine derivatives with omega-methoxyPEG oxiranylmethyl ethers. Constructs carrying 1-3 MeOPEG units and 0, 2 or 4 N-methyl groups have been prepared by this method. H2N(CH2)3NBoc(CH2)3NBoc(CH2)3NHBoc was prepared efficiently by mono-trifluoroacetylation of thermine, attachment of Boc and removal of the trifluoroacetyl group in one pot. A similar process gave H2N(CH2)3NBoc(CH2)3NBoc(CH2)3NH2. BocMeN(CH2)3NHMe was alkylated by 1,3-dibromopropane to give BocMeN(CH2)3NMe(CH2)3NMe(CH2)3NMeBoc. A cyanoethylation/reduction sequence extended H2N(CH2)3NBoc(CH2)3NBoc(CH2)3NH2 to give H2N(CH2)3NBoc(CH2)3NBoc(CH2)3NBoc(CH2)3NBoc(CH2) 3NH2, which was converted to its mono- and di-MeOPEG550 derivatives. Deprotection gave the linear polyamine MeOPEG constructs. A branched triamine-poly(ethylene glycol) construct was prepared by acylation of (BocHN(CH2)3)2N(CH2)3NH2 with omega-methoxyPEG 550 chloroformate, followed by deprotection. A cyanoethylation/reduction/protection sequence from (H2N(CH2)3)2 N(CH2)3NHBoc gave a protected pentamine. Alkylation with Br(CH2)5CONH(CH2)2NHBoc, deprotection, acylation with MeOPEG chloroformate and deprotection gave a pentamine MeOPEG construct in which the MeOPEG is attached through a linker to the central amine. The linear hexamine construct carrying MeOPEG550 at only one terminus was the most effective DNA-interactive member of the two series in an ethidium displacement assay and was effective in delivering a reporter gene to RIF-1 tumours.
Subject(s)
DNA/metabolism , Drug Delivery Systems , Polyamines/metabolism , Polyethylene Glycols/metabolism , Animals , Binding, Competitive , Cell Nucleus/metabolism , Chloramphenicol O-Acetyltransferase/genetics , Chloramphenicol O-Acetyltransferase/metabolism , DNA/administration & dosage , DNA/ultrastructure , Drug Compounding , Ethidium/metabolism , Female , Genes, Reporter/genetics , Humans , Mice , Mice, Inbred C3H , Polyamines/chemical synthesis , Polyethylene Glycols/chemical synthesis , Sarcoma, Experimental/genetics , Sarcoma, Experimental/metabolism , Surface-Active Agents/chemical synthesis , Surface-Active Agents/chemistry , Surface-Active Agents/metabolism , Transfection/methodsABSTRACT
Acid-catalysed condensation of 5-nitrofuran-2-carboxaldehyde with 1-phenylethane-1,2-diol and with 2,2-dimethyl-1-phenylproane-1,3-diol in boiling benzene gave the expected cyclic nitrofuranyl acetals. Biomimetic reduction of these acetals with sodium borohydride in the presence of palladium triggered release of the parent diols. Thus these nitrofuran acetals may have potential for applications as prodrugs for selective release of diol-containing drugs in hypoxic solid tumours.
