Winkler Johannes, Urban Ernst, Noe Christian R
Department of Medicinal Chemistry, Universität Wien, Althanstrasse 14, 1090 Wien, Austria.
Bioconjug Chem. 2005 Jul-Aug;16(4):1038-44. doi: 10.1021/bc049729d.
A new method for synthesizing oligonucleotide peptide conjugates by an in-line approach is presented. A phosphorothioate oligonucleotide with the sequence of bcl-2 targeted Oblimersen by employing a modified 2'-amino-2'-desoxy-uridine nucleotide bearing a succinyl linker at the 2' position was prepared. The carboxyl group was protected for solid-phase synthesis as the benzyl ester. Ester cleavage was afforded by a phase transfer reaction using palladium nanoparticles as catalyst and cyclohexadiene as hydrogen donor. Short tails of up to three lysyl residues were conjugated to the oligonucleotide by an inverse stepwise peptide synthesis. The conjugates were characterized by HPLC, mass spectrometry, and circular dichroism. Influence of lysyl tails on CD spectra were minimal. Melting profiles revealed only minimal destabilizing effects on duplexes by conjugation of peptides.
本文介绍了一种通过在线方法合成寡核苷酸肽缀合物的新方法。制备了一种硫代磷酸酯寡核苷酸，其序列为靶向bcl-2的奥布利默森，采用在2'位带有琥珀酰接头的修饰2'-氨基-2'-脱氧尿苷核苷酸。羧基作为苄酯被保护用于固相合成。使用钯纳米颗粒作为催化剂和环己二烯作为氢供体，通过相转移反应进行酯裂解。通过反向逐步肽合成将最多三个赖氨酰残基的短尾巴缀合到寡核苷酸上。通过高效液相色谱、质谱和圆二色性对缀合物进行了表征。赖氨酰尾巴对圆二色光谱的影响最小。熔解曲线显示，肽的缀合对双链体只有最小的去稳定作用。
