The DiPPro Approach: Synthesis, Hydrolysis, and Antiviral Activity of Lipophilic d4T Diphosphate Prodrugs
The DiPPro Approach: Synthesis, Hydrolysis, and Antiviral Activity of Lipophilic d4T Diphosphate Prodrugs
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DOI：
10.1002/cmdc.201300500
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发表时间：
2014-04-01
期刊：
影响因子：
3.4
通讯作者：
Meier, Chris
中科院分区：
文献类型：
--
作者：
Schulz, Tilmann;Balzarini, Jan;Meier, Chris
提出了双（酰氧基苄基）磷酸酯形式的核苷二磷酸（NDP）的β-磷酸基团的生物可逆保护。为了研究这些潜在的 NDP 前药（DiPPro 药物）的结构-活性关系，合成了一系列带有不同长度的脂肪酸和 d4T 作为模型核苷的 DiPPro 化合物。为了合成亲脂修饰的二磷酸基团，使预先形成的亚磷酰胺与核苷酸反应，随后氧化-P-III部分。在不同的介质（如磷酸盐缓冲液 (pH7.3) 或 CEM 细胞提取物）中研究了这些前药的化学和酶稳定性。在所有介质中，水解速率明显取决于酰基部分，并且随着烷基链长度的增加而降低。由于存在酶催化裂解，这些化合物在细胞提取物中的半衰期明显低于在 pH7.3 磷酸盐缓冲液中的半衰期。在所有介质中，DiPPro 化合物释放 d4T 二磷酸盐 (d4TDP) 作为除 d4TMP 之外的主要产物。在抗病毒测定中，这些化合物在野生型 CEM/0 细胞中被证明至少与 d4T 一样有效对抗 HIV-1 和 2。作为概念证明，具有较长酰基残基的化合物在胸苷激酶缺陷细胞（CEM/TK-）中表现出非常好的抗 HIV 活性，表明它们具有穿透细胞膜和递送磷酸化代谢物的能力。
Bioreversible protection of the -phosphate group of nucleoside diphosphates (NDPs) as bis(acyloxybenzyl)phosphate esters is presented. To investigate the structure-activity relationship of these potential NDP prodrugs (DiPPro drugs) a series of DiPPro compounds was synthesized bearing fatty acids of various lengths and d4T as a model nucleoside. For synthesis of the lipophilically modified diphosphate group, preformed phosphoramidites were allowed to react with nucleotides, and the -P-III moiety was subsequently oxidized. The chemical and enzymatic stability of these prodrugs was studied in different media such as phosphate buffer (pH7.3) or CEM cell extracts. In all media, the hydrolysis rate was clearly dependent on the acyl moiety and decreased with increasing alkyl chain length. The compounds showed a markedly lower half-life in cell extracts than in pH7.3 phosphate buffer due to the presence of enzyme-catalyzed cleavage. In all media, the DiPPro compounds released d4T diphosphate (d4TDP) as the main product beside d4TMP. In antiviral assays, the compounds proved to be at least as potent as d4T against HIV-1 and 2 in wild-type CEM/0 cells. As a proof of concept, compounds with longer acyl residues showed very good anti-HIV activities in thymidine-kinase-deficient cells (CEM/TK-), indicating their ability to penetrate cell membranes and the delivery of phosphorylated metabolites.