Synthesis of AZT 5'-triphosphate mimics and their inhibitory effects on HIV-1 reverse transcriptase.
Synthesis of AZT 5'-triphosphate mimics and their inhibitory effects on HIV-1 reverse transcriptase.
复制标题
AZT 5-三磷酸模拟物的合成及其对 HIV-1 逆转录酶的抑制作用。
DOI：
10.1021/jm040116w
复制
发表时间：
2004
影响因子：
7.3
通讯作者：
P. Dan Cook
中科院分区：
文献类型：
--
作者：
Guangyi Wang;N. Boyle;Fu Chen;V. Rajappan;P. Fagan;Jennifer L. Brooks;Tiffany Hurd;Janet M. Leeds;V. Rajwanshi;Yi Jin;M. Prhavc;T. W. Bruice;P. Dan Cook
为了寻找活性核苷5‘-三磷酸模拟物，我们合成了一系列AZT三磷酸模拟物(AZT P3ms)，并评价了它们对HIV-1逆转录酶的抑制作用以及它们在胎牛血清和CEM细胞提取物中的稳定性。AZT与2-氯-4H-1，3，2-苯二氧磷-4-酮反应，然后用焦磷酸盐类似物处理亚磷酸盐中间体2，得到环状三磷酸中间体4b-4f，再经过硼化和随后的水解，以中等到较好的产率得到AZT 5‘-α-硼基-β，伽马桥修饰的三磷酸盐6b-6f。环状中间体4d与碘反应，然后用一系列亲核试剂处理，得到了AZT 5‘-β，伽马-二氟亚甲基-伽马取代三磷酸盐(7b-7i)。合成了几种不同类型的含α-P-硫代(或二硫代)和β-二氟亚甲基(13，14)，α，β-二氟甲基和伽马-对甲基(或苯基)(15，16)，以及α-硼基-β，伽马-二氟甲基和伽马-O-甲基/苯基(11，12)的AZT P3M。以聚(A)均聚物为模板，用荧光法测定了化合物作为HIV-1逆转录酶抑制剂的有效性。许多AZT P3M对HIV-1逆转录酶有很强的抑制作用。在三磷酸的β，伽马桥的修饰使AZT P3MS 6b-6f具有不同的活性(K(I)从9.5到>；>；500 nm)，而在三磷酸的α，β桥的修饰导致弱的AZT P3M抑制剂。结果表明，AZT P3M和AZT三磷酸均为底物抑制剂。最具活性的化合物，AZT 5‘-α-R(P)()-硼基-β-二氟亚甲基三磷酸(AZT 5’-Alphab-BetagammaCF(2)TP)(6d-i)，与AZT三磷酸一样有效，K(I)()值为9.5 NM，在血清和细胞提取物中的稳定性至少是AZT三磷酸的20倍。因此，首次确定了一种活性高、稳定性好的核苷三磷酸模拟物，有望作为一种新型的抗病毒药物。5‘-α-硼基-β，伽马-(二氟亚甲基)三磷酸是一种很有前景的三磷酸盐类化合物，有望在核苷酸类抗病毒药物的开发中发挥重要作用。
In search of active nucleoside 5'-triphosphate mimics, we have synthesized a series of AZT triphosphate mimics (AZT P3Ms) and evaluated their inhibitory effects on HIV-1 reverse transcriptase as well as their stability in fetal calf serum and in CEM cell extracts. Reaction of AZT with 2-chloro-4H-1,3,2-benzodioxaphosphorin-4-one, followed by treatment of the phosphite intermediate 2 with pyrophosphate analogues, yielded the cyclic triphosphate intermediates 4b-4f, which were subjected to boronation and subsequent hydrolysis to give AZT 5'-alpha-borano-beta,gamma-bridge-modified triphosphates 6b-6f in moderate to good yields. Reaction of the cyclic intermediate 4d with iodine, followed by treatment with a series of nucleophiles, afforded the AZT 5'-beta,gamma-difluoromethylene-gamma-substituted triphosphates (7b-7i). Several different types of AZT P3Ms containing alpha-P-thio (or dithio) and beta,gamma-difluoromethylene (13,14), alpha,beta-difluoromethylene and gamma-P-methyl(or phenyl) (15,16), and alpha-borano-beta,gamma-difluoromethylene and gamma-O-methyl/phenyl (11,12) were also synthesized. The effectiveness of the compounds as inhibitors of HIV-1 reverse transcriptase was determined using a fluorometric assay and a poly(A) homopolymer as a template. A number of AZT P3Ms exhibited very potent inhibition of HIV-1 reverse transcriptase. Modifications at the beta,gamma-bridge of triphosphate rendered the AZT P3Ms 6b-6f with varied activities (K(i) from 9.5 to >>500 nM) while modification at the alpha,beta-bridge of triphosphate led to weak AZT P3M inhibitors. The results imply that the AZT P3Ms were substrate inhibitors, as is AZT triphosphate. The most active compound, AZT 5'-alpha-R(p)()-borano-beta,gamma-(difluoromethylene)triphosphate (AZT 5'-alphaB-betagammaCF(2)TP) (6d-I), is as potent as AZT triphosphate with a K(i)() value of 9.5 nM and at least 20-fold more stable than AZT triphosphate in the serum and cell extracts. Therefore, for the first time, a highly active and stable nucleoside triphosphate mimic has been identified, which is potentially useful as a new type of antiviral drug. The promising triphosphate mimic, 5'-alpha-borano-beta,gamma-(difluoromethylene)triphosphate, is expected to be valuable to the discovery of nucleotide mimic antiviral drugs.