Kramata P, Votruba I, Otová B, Holý A
Institute of Organic Chemistry and Biochemistry, Academy of Sciences of The Czech Republic, Prague, The Czech Republic.
Mol Pharmacol. 1996 Jun;49(6):1005-11.
Based on the powerful virostatic potency and cytostatic activity of adenine, 2,6-diaminopurine, and guanine derivatives of acyclic phosphonate nucleotide analog (S)-1-(3-hydroxy-2-phosphonomethoxypropyl) and 9-(2-phosphonomethoxyethyl) series, we examined the inhibitory potencies of their diphosphates [(S)-9-(3-hydroxy-2-phosphonomethoxypropyl)adenine diphosphate (HPMPApp), 9-(2-phosphonomethoxyethyl)adenine diphosphate, 9-(2-phosphonomethoxyethyl)-2,6-diaminopurine diphosphate (PMEDAPpp), and 9-(2-phosphonomethoxyethyl)guanine diphosphate, analogs of nucleoside 5'-triphosphates] toward cellular DNA polymerases alpha, delta, and epsilon (isolated from tumors of T cell spontaneous acute lymphoblastic leukemia in Sprague-Dawley inbred rats). Kinetic measurements (K(m), K(i), and V(max)) of synthetic homopolymeric template primers have shown that HPMPApp is a selective and potent inhibitor of polymerase epsilon, whereas PMEDAPpp strongly inhibits polymerase delta. These two compounds may be useful for elucidating the roles of polymerases delta and epsilon. Of the nucleotide analogs tested, 9-(2-phosphonomethoxyethyl) guanine diphosphate is the most efficient inhibitor of polymerases alpha and epsilon, whereas the diphosphate of 9-(2-phosphonomethoxyethyl) adenine, the therapeutically important agent adefovir, inhibits polymerases alpha and epsilon relatively poorly and exerts only moderate inhibition of polymerase delta. These data are quite consistent with previously reported cytostatic activity of these nucleotide analogs. All of the enzymes studied catalyze the incorporation of 9-(2-phosphonomethoxyethyl)adenine, 9-(2-phosphonomethoxyethyl)-2, 6-diaminopurine, and (S)-9-(3-hydroxy-2-phosphonomethoxypropyl)adenine into DNA chain. 9-(2-Phosphonomethoxyethyl)adenine diphosphate and PMEDAPpp were confirmed to be DNA chain terminators. On the other hand, HPMPApp formed poly(dT)/oligo(dA(18)-[(S)-9-(3-hydroxy-2-phosphonomethoxypropyl)a denine]2-4 structures.
基于无环膦酸核苷酸类似物（S）-1-（3-羟基-2-膦酰甲氧基丙基）和9-（2-膦酰甲氧基乙基）系列的腺嘌呤、2,6-二氨基嘌呤和鸟嘌呤衍生物强大的病毒静止效力和细胞生长抑制活性，我们检测了它们的二磷酸酯[（S）-9-（3-羟基-2-膦酰甲氧基丙基）腺嘌呤二磷酸（HPMPApp）、9-（2-膦酰甲氧基乙基）腺嘌呤二磷酸、9-（2-膦酰甲氧基乙基）-2,6-二氨基嘌呤二磷酸（PMEDAPpp）和9-（2-膦酰甲氧基乙基）鸟嘌呤二磷酸，核苷5'-三磷酸类似物]对细胞DNA聚合酶α、δ和ε（从斯普拉格-道利近交系大鼠的T细胞自发性急性淋巴细胞白血病肿瘤中分离）的抑制效力。对合成的均聚模板引物的动力学测量（K(m)、K(i)和V(max)）表明，HPMPApp是聚合酶ε的选择性强效抑制剂，而PMEDAPpp强烈抑制聚合酶δ。这两种化合物可能有助于阐明聚合酶δ和ε的作用。在所测试的核苷酸类似物中，9-（2-膦酰甲氧基乙基）鸟嘌呤二磷酸是聚合酶α和ε最有效的抑制剂，而治疗上重要的药物阿德福韦的9-（2-膦酰甲氧基乙基）腺嘌呤二磷酸对聚合酶α和ε的抑制相对较弱，对聚合酶δ仅产生中度抑制。这些数据与先前报道的这些核苷酸类似物的细胞生长抑制活性相当一致。所有研究的酶都催化将9-（2-膦酰甲氧基乙基）腺嘌呤、9-（2-膦酰甲氧基乙基）-2,6-二氨基嘌呤和（S）-9-（3-羟基-2-膦酰甲氧基丙基）腺嘌呤掺入DNA链。9-（2-膦酰甲氧基乙基）腺嘌呤二磷酸和PMEDAPpp被证实是DNA链终止剂。另一方面，HPMPApp形成了聚（dT）/寡聚（dA(18)-[(S)-9-（3-羟基-2-膦酰甲氧基丙基）腺嘌呤]2-4结构。
