Zarytova V F, Sergeyev D S, Godovikova T S
Institute of Bioorganic Chemistry, Siberian Division of the Russian Academy of Sciences, Russia.
Bioconjug Chem. 1993 May-Jun;4(3):189-93. doi: 10.1021/bc00021a001.
A method for coupling bleomycin A5 to oligonucleotides is proposed. The reaction was carried out between an amino group of a spermidine residue of the Cu(II) complex of bleomycin A5 (Cu(II)Blm-RH) and a 5'-phosphate group of the oligonucleotides d(pCCAAACA) (I), d(pCGTCCTC) (II), d(pT)16 (III), d(pCAAACA) (IV), and d(pGCCAAACA) (V) activated with a mixture of triphenylphosphine and 2,2'-dipyridyl disulfide in the presence of 4-(N,N-dimethylamino)pyridine 1-oxide. The yields of the products Cu(II)Blm-R-d(pCCAAACA) (Ia), Cu(II)Blm-R-d(pCGTCCTC) (IIa), Cu(II)Blm-R-d(pT)16 (IIIa), Cu(II)Blm-R-d(pCAAACA) (IVa), and Cu(II)Blm-R-d(pGCCAAACA) (Va) were 60-80%. After removal of the Cu(II) ion from the bleomycin A5 oligonucleotide derivatives Ia-IIIa, compounds Ib-IIIb were obtained. Compounds Ia, IVa, Va, and Ib-IIIb were further used for modification of the target d(pTGTTTGGCGAAGGA) in the presence of Fe(II) ions and 2-mercaptoethanol. Site-specific cleavage of the target by Blm coupled to complementary oligonucleotides was demonstrated. It was shown that efficiency and position of cleavage of the complementary reagents Ia, Ib, IVa, and Va are determined by their oligonucleotide part while the action of thenoncomplementary reagents IIb and IIIb was similar to that of the free antibiotic.
提出了一种将博来霉素A5与寡核苷酸偶联的方法。该反应在博来霉素A5的铜（II）配合物（Cu（II）Blm-RH）的亚精胺残基的氨基与用三苯基膦和2,2'-二吡啶二硫化物的混合物在4-（N，N-二甲基氨基）吡啶1-氧化物存在下活化的寡核苷酸d（pCCAAACA）（I）、d（pCGTCCTC）（II）、d（pT）16（III）、d（pCAAACA）（IV）和d（pGCCAAACA）（V）的5'-磷酸基团之间进行。产物Cu（II）Blm-R-d（pCCAAACA）（Ia）、Cu（II）Blm-R-d（pCGTCCTC）（IIa）、Cu（II）Blm-R-d（pT）16（IIIa）、Cu（II）Blm-R-d（pCAAACA）（IVa）和Cu（II）Blm-R-d（pGCCAAACA）（Va）的产率为60 - 80%。从博来霉素A5寡核苷酸衍生物Ia - IIIa中除去铜（II）离子后，得到化合物Ib - IIIb。化合物Ia、IVa、Va和Ib - IIIb在亚铁离子和2 - 巯基乙醇存在下进一步用于修饰靶标d（pTGTTTGGCGAAGGA）。证明了与互补寡核苷酸偶联的博来霉素对靶标的位点特异性切割。结果表明，互补试剂Ia、Ib、IVa和Va的切割效率和位置由其寡核苷酸部分决定，而非互补试剂IIb和IIIb的作用与游离抗生素相似。
