Molecular designing of nucleoside analogues for inhibitor of telomerase

核苷和相应的5′-三磷酸作为人类端粒酶的潜在选择性抑制剂的分子设计已被考虑。首先，我们详细研究了拉伸PCR法测定HeLa细胞端粒酶。所得结果可用于后续的酶动力学分析，包括Lineweaver-Burk图。与HIV-1逆转录酶相比，天然底物dGTP和dTTP的对映体L-dGTP和L-dTIP对端粒酶的抑制作用较弱。这表明端粒酶可以严格识别底物的对映体，另一方面，一个5-苯基取代的dUTP类似物对端粒酶表现出较强的亲和力，因此该酶更倾向于疏水和无碱基取代基。在此基础上，合成了5-叠氮多甲基-、氨基多甲基-、羟甲基-和甲氧基甲基-等dTTP衍生物，并制备了具有不同糖基的糖基取代基脲基核苷酸。作为端粒酶的另一来源，对樱桃鲑鱼进行了研究，发现端粒酶活性很强。这些发现可用于进一步研究该酶在蛋白质结构和功能水平上的作用。

Molecular designing of nucleosides and corresponding 5'-triphosphates as potential selective inhibitor for human telomerase have been considered.First, we have examined the stretch PCR method for determination of HeLa cell telomerase in detail. The results obtained could be used in following experiments including enzyme kinetic analysis by Lineweaver-Burk plots. L-dGTP and L-dTIP, enantiomers of natural substrates dGTP and dTTP, showed inhibitory effect on telomerase in lesser extent when compared with HIV-1 reverse transcriptase. This result clearly indicated that telomerase can strictly recognize enantiomer for its substrate, On the other hand, a 5-styryl-substituted dUTP analogue has shown strong affinity to the telomerase, thus this enzyme was prefer hydrophobic and steric substituent no base moieties. Based on these findings, several dTTP analogues such as 5-azidomethyl-, aminomethyl-, hydroxymethyl- and methoxymethly-derivatives were synthesized, additionally, sugar replacement lyxofuranosyl nucleotides having various aglycon moieties were also prepared. As the other source for telomerase, cherry salmon tests were investigated and found strong telomerase activity. These findings should be applicable to further studies of the enzyme on the level of protein structure and function.