Maruenda H, Tomasz M
Department of Chemistry, Hunter College, City University of New York, New York 10021, USA.
Bioconjug Chem. 1996 Sep-Oct;7(5):541-4. doi: 10.1021/bc960054r.
Oligodeoxyribonucleotides (ODNs) conjugated with mitomycin C (MC) via (-CH2-)n tethers of different lengths (n = 6, 12) to their terminal 5'-phosphate were synthesized, and their interaction with target complementary single-stranded DNA oligonucleotides was investigated. MC, a clinically used natural anticancer drug, is known to act as a bioreductive alkylating agent of duplex DNA with a remarkable preference for 5'-d(CG) sequences. The usual enzymatic bioreductive techniques known to trigger MC to alkylate DNA were employed in the reaction between the MC-oligonucleotide conjugates and their targets radiolabeled by 32P at their 5'-phosphate. A slow-moving radiolabeled product, detected by polyacrylamide gel electrophoresis using phosphorimaging techniques, was obtained in 15-25% yield with complementary DNA as target. Formation of these products was dependent upon complementary duplex formation. Evidence is presented that the DNA target is alkylated by the mitomycin C moiety of the ODN conjugate at the 2-amino group of a guanine base. These findings suggest that the MC-ODN conjugates may be useful specific inhibitors of cellular or viral gene expression. To our knowledge this is the first report on ODN conjugates of a reductively activated drug of known therapeutic value.
合成了通过不同长度（n = 6, 12）的(-CH2-)n连接链与丝裂霉素C（MC）连接至其末端5'-磷酸的寡脱氧核糖核苷酸（ODN），并研究了它们与靶标互补单链DNA寡核苷酸的相互作用。MC是一种临床使用的天然抗癌药物，已知其作为双链DNA的生物还原烷基化剂，对5'-d(CG)序列有显著偏好。在MC-寡核苷酸缀合物与其在5'-磷酸处用32P进行放射性标记的靶标之间的反应中，采用了已知能触发MC使DNA烷基化的常规酶促生物还原技术。以互补DNA为靶标，通过使用磷光成像技术的聚丙烯酰胺凝胶电泳检测到一种迁移缓慢的放射性标记产物，产率为15 - 25%。这些产物的形成取决于互补双链的形成。有证据表明，DNA靶标在鸟嘌呤碱基的2-氨基处被ODN缀合物的丝裂霉素C部分烷基化。这些发现表明，MC-ODN缀合物可能是细胞或病毒基因表达的有用特异性抑制剂。据我们所知，这是关于具有已知治疗价值的还原激活药物的ODN缀合物的首次报道。
