Ede N J, Tregear G W, Haralambidis J
Howard Florey Institute of Experimental Physiology and Medicine, University of Melbourne, Parkville, Victoria, Australia.
Bioconjug Chem. 1994 Jul-Aug;5(4):373-8. doi: 10.1021/bc00028a016.
Oligonucleotide-peptide hybrids have potential for use as antisense inhibitors of gene expression, with the peptide helping to increase the intracellular concentration of the active oligonucleotide. The preparation of such hybrids can be achieved by the coupling of thiol-derivatized oligonucleotides with maleimido-peptides. We have developed reliable methods for preparing 5'-thiol oligonucleotides in good yields using phosphoramidite chemistry and coupling 6-(tritylthio)hexyl phosphoramidite as the 5'-terminal residue. The use of highly pure thiol phosphoramidite as well as a manual iodine treatment after this coupling were found to be important. Oligonucleotide-peptide hybrids were prepared in high yield (85%) by reacting freshly purified 5'-thiol oligonucleotides with peptides derivatized at their N-terminus with a maleimido functionality.
寡核苷酸 - 肽杂合体有潜力用作基因表达的反义抑制剂，其中肽有助于提高活性寡核苷酸的细胞内浓度。此类杂合体的制备可通过将硫醇衍生化的寡核苷酸与马来酰亚胺肽偶联来实现。我们已经开发出可靠的方法，使用亚磷酰胺化学以高收率制备5'-硫醇寡核苷酸，并将6-(三苯甲基硫基)己基亚磷酰胺作为5'-末端残基进行偶联。发现使用高纯度硫醇亚磷酰胺以及偶联后进行手动碘处理很重要。通过使新鲜纯化的5'-硫醇寡核苷酸与在其N-末端衍生有马来酰亚胺官能团的肽反应，以高收率（85%）制备了寡核苷酸 - 肽杂合体。
