Synthesis of New NNRTLs for the Treatment of AIDS

描述（由申请人提供）： 目前需要新的非核苷HIV-1逆转录酶抑制剂（NNRTI），其：1）具有比现有NNRTI更低的毒性，2）具有独特的抗性突变谱，并且对对现有NNRTI具有抗性的突变型逆转录酶保持活性，3）当与其他抗HIV药物组合使用时，具有抑制抗性病毒株出现的能力，4）与其他抗HIV剂组合具有协同抗HIV活性，5）比现有亚当斯更稳定，因此具有增强的生物利用度，6）对各种HIV-1菌株具有广泛的活性，和7）对RT具有高亲和力，因此具有高抗HIV效力的潜力。我们的研究小组最近报道了一系列新的烯基二芳基甲烷（ADAM）NNRTIs的设计和合成，这些NNRTIs是HIV-1致细胞病变效应的有效抑制剂。尽管一些亚当斯在低纳摩尔浓度下抑制CEM-SS细胞培养物中HIV-1 RF的致细胞病变作用，但亚当斯的潜在治疗效用受到容易被血浆酯酶水解的三个甲基酯部分的存在的损害。因此，本项目的主要目标是找到这三种不稳定酯的合适替代品，这些酯将是代谢稳定的，并且还将保留母体化合物的有效抗HIV活性。初步研究表明，恶唑烷酮和甲基醚取代甲酯产生的活性很有希望，并提出了各种其他代谢稳定的部分。这些包括乙基酮、异丁烯、烯醇醚、醚、乙烯基酮、正丙基、α，α-二氟酮、二氟烯醇醚、异丙基异丁烯、二氟异丁烯、亚氨酰氟、酰胺、硫酯、硫酸酯和二硫酯替代物。合成提出了含有这些结构简单的甲基酯取代的亚当斯。将在人血浆中研究新型亚当斯的水解稳定性。此外，新亚当斯的抗HIV活性将在多种生物系统中测定。将在细胞培养物中测定化合物作为多种HIV-1株致细胞病变效应抑制剂的效力。还将研究化合物在未感染淋巴细胞中的细胞毒性。将使用野生型和突变蛋白在无细胞系统中建立亚当斯的酶抑制活性。作用机制研究将包括添加时间（时程）研究以及在采用代表病毒复制周期各个阶段的靶标的许多测定中对化合物进行检查。将确定化合物与现有抗HIV剂的协同活性。将研究亚当斯与NNRTI耐药病毒的活性。新亚当斯的水溶解度将被准确地测量。

DESCRIPTION (provided by applicant): A current need exists for novel non-nucleoside HIV-1 reverse transcriptase inhibitors (NNRTIs) that: 1) have lower toxicities than the existing NNRTIs, 2) have unique resistance mutation profiles and remain active against mutant reverse transcriptases that are resistant to the existing NNRTIs, 3) have the ability to suppress the emergence of resistant viral strains when used in combination with other anti-HIV agents, 4) have synergistic anti-HIV activity in combination with other anti-HIV agents, 5) are metabolically more stable than the existing ADAMs and therefore have enhanced bioavailabilities, 6) have a wide range of activity vs. various HIV-1 strains, and 7) have high affinities for RT, and therefore have the potential for high anti-HIV potency. Our research group has recently reported the design and synthesis of a novel series of alkenyldiarylmethane (ADAM) NNRTIs that are potent inhibitors of the cytopathic effect of HIV-1. Although some of the ADAMs inhibit the cytopathic effect of HIV-1RF in CEM-SS cell culture at low nanomolar concentrations, the potential therapeutic utility of the ADAMs is compromised by the presence of three methyl ester moieties that are readily hydrolyzed by plasma esterases. Consequently, the main goal of the present project is to find suitable replacements for these three labile esters that will be metabolically stable and will also retain the potent anti-HIV activity of the parent compound. Preliminary studies have shown promising activity resulting from oxazolidinone and methyl ether replacements of the methyl esters, and a variety of additional metabolically stable moieties are proposed. These include ethyl ketone, isobutylene, enol ether, ether, vinyl ketone, n-propyl, alpha, alpha -difluoroketone, difluoroenol ether, tetrafluoroisobutylene, difluoroisobutylene, imidoyl fluoride, amide, thioester, thionoester, and dithioester replacements. Syntheses are proposed for ADAMs containing each of these structurally simple methyl ester replacements. The hydrolytic stabilities of the new ADAMs will be investigated in human plasma. In addition, the anti-HIV activities of the new ADAMs will be determined in a variety of biological systems. The potencies of the compounds as inhibitors of the cytopathic effect of a variety of HIV-1 strains will be determined in cell culture. The cytotoxicities of the compounds in uninfected lymphocytes will also be investigated. The enzyme inhibitory activities of the ADAMs will be established in cell-free systems using both wild type and mutant proteins. Mechanism of action studies will include both time-of-addition (time course) studies as well as the examination of the compounds in a number of assays employing targets that represent various stages in the replication cycle of the virus. The synergistic activities of the compounds with existing anti-HIV agents will be established. The activities of the ADAMs vs. NNRTI resistant viruses will be investigated. The aqueous solubilities of the new ADAMs will be measured accurately.