Synthesis and characterisation of polyamine-poly(ethylene glycol) constructs for DNA binding and gene delivery

Shane W. Garrett, Owen R. Davies, David A. Milroy, Pauline J. Wood, Colin W. Pouton, Michael D. Threadgill

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34 Citations (Scopus)

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 ω-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 ω-methoxyPEG 550 chloroformate, followed by deprotection. A cyanoethylation/reduction/protection sequence from (H2N(CH2)3)2N(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. (C) 2000 Elsevier Science Ltd.

Original languageEnglish
Pages (from-to)1779-1797
Number of pages19
JournalBioorganic & Medicinal Chemistry
Volume8
Issue number7
DOIs
Publication statusPublished - 2000
Externally publishedYes

Keywords

  • Animals Binding, Competitive Cell Nucleus/metabolism Chloramphenicol O-Acetyltransferase/genetics/metabolism DNA/administration & dosage/*metabolism/ultrastructure Drug Compounding *Drug Delivery Systems Ethidium/metabolism Female Genes, Reporter/genetics Humans Mice Mice, Inbred C3H Polyamines/chemical synthesis/*metabolism Polyethylene Glycols/chemical synthesis/*metabolism Sarcoma, Experimental/genetics/metabolism Surface-Active Agents/chemical synthesis/chemistry/metabolism Transfection/methods

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