Preclinical evaluation of a mercaptobenzamide and its prodrug for NCp7-targeted inhibition of human immunodeficiency virus.
Preclinical evaluation of a mercaptobenzamide and its prodrug for NCp7-targeted inhibition of human immunodeficiency virus.
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DOI：
10.1016/j.antiviral.2016.08.022
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发表时间：
2016-10
影响因子：
7.6
通讯作者：
Appella DH
中科院分区：
文献类型：
--
作者：
Hartman TL;Yang L;Helfrick AN;Hassink M;Shank NI;George Rosenker K;Scerba MT;Saha M;Hughes E;Wang AQ;Xu X;Gupta P;Buckheit RW Jr;Appella DH
尽管有效使用高效抗逆转录病毒疗法可以抑制感染个体中的病毒产生，但它并不能消除避难所中携带病毒的细胞中的感染和低水平病毒产生。因此，继续寻找具有独特和不同的 HIV 抑制机制的新型抗逆转录病毒药物仍然至关重要，而能够降低已感染 HIV 的细胞产生病毒水平的化合物，而不是预防从头感染，将大有裨益。巯基苯甲酰胺 (MDH-1-38) 及其前药 (NS1040) 正在开发中，作为针对 HIV 核衣壳锌指的潜在治疗化合物。在酯酶存在下，NS1040 被设计为转化为具有抗病毒活性的 MDH-1-38。虽然我们假设 NS1040 在所有实验中都会快速转化为 MDH-1-38，但同时对这两种化合物进行了测试，以确定前药的存在是否会影响抗病毒活性或作用机制。这两种化合物针对人类 PBMC 和单核巨噬细胞中的一组 HIV-1 临床分离株进行了评估，得出的 EC50 值范围为 0.7 至 13 μM，且最高 100 μM 没有毒性。在添加血清蛋白的情况下，MDH-1-38 和 NS1040 在人 PBMC 中保持同样的活性，并且针对对逆转录酶、整合酶或蛋白酶抑制剂具有抗性的 HIV-1 分离株。基于细胞和生化机制的抗病毒作用测定表明，MDH-1-38 和 NS1040 分别在浓度为 15 和 50 μM 时具有杀病毒作用。在 CEM-SS 和人 PBMC 中，通过降低子代病毒感染性（化合物浓度大于 2 μM），多次传代中 HIV 的细胞间传播显着减少。 MDH-1-38 或 NS1040 与 FDA 批准的其他 HIV 药物联合使用可产生协同抗病毒相互作用，且没有抗病毒拮抗或协同毒性的证据。使用连续剂量递增来尝试选择对 MDH-1-38 和 NS1040 具有抗性的 HIV 毒株。病毒在数次传代中未能在以增加的化合物浓度处理的细胞中复制，这与病毒灭活化合物的拟议作用机制一致。经过 14 次传代，尚未实现对化合物的耐药性。大多数具有针对病毒蛋白的抗病毒作用机制的 HIV 抑制剂会在 14 代内选择出耐药病毒。这些研究表明，这些 NCp7 靶向化合物代表了新的有效抗 HIV 候选药物，可以有效地与所有已批准的抗 HIV 药物联合使用。具有抗 HIV-1 活性，在 PBMC 和单核细胞中的 EC50 范围为 0.7 至 16 μM。有效对抗耐药 HIV-1，EC50 与野生型病毒相似。作用机制研究表明 MDH-1-38 是一种杀病毒剂。与 HIV 药物的组合产生了协同作用的加和性，且没有细胞毒性。传代14次后尚未达到耐药性，导致病毒有缺陷。
Although the effective use of highly active antiretroviral therapy results in the suppression of virus production in infected individuals, it does not eliminate the infection and low level virus production in cells harboring virus in sanctuary sites. Thus, the continued search for new antiretroviral agents with unique and different mechanisms of HIV inhibition remains critical, and compounds that can reduce the level of virus production from cells already infected with HIV, as opposed to preventing de novo infection, would be of great benefit. A mercaptobenzamide (MDH-1-38) and its prodrug (NS1040) are being developed as potential therapeutic compounds targeting the zinc finger of HIV nucleocapsid. In the presence of esterase enzymes, NS1040 is designed to be converted to MDH-1-38 which has antiviral activity. While we presume that NS1040 is rapidly converted to MDH-1-38 in all experiments, the two compounds were tested side-by-side to determine whether the presence of a prodrug affects the antiviral activity or mechanism of action. The two compounds were evaluated against a panel of HIV-1 clinical isolates in human PBMCs and monocyte-macrophages and yielded EC50 values ranging from 0.7 to 13 μM with no toxicity up to 100 μM. MDH-1-38 and NS1040 remained equally active in human PBMCs in the presence of added serum proteins as well as against HIV-1 isolates resistant to reverse transcriptase, integrase or protease inhibitors. Cell-based and biochemical mechanism of antiviral action assays demonstrated MDH-1-38 and NS1040 were virucidal at concentrations of 15 and 50 μM, respectively. Cell to cell transmission of HIV in multiple passages was significantly reduced in CEM-SS and human PBMCs by reducing progeny virus infectivity at compound concentrations greater than 2 μM. The combination of either MDH-1-38 or NS1040 with other FDA-approved HIV drugs yielded additive to synergistic antiviral interactions with no evidence of antiviral antagonism or synergistic toxicity. Serial dose escalation was used in attempts to select for HIV strains resistant to MDH-1-38 and NS1040. Virus at several passages failed to replicate in cells treated at increased compound concentrations, which is consistent with the proposed mechanism of action of the virus inactivating compounds. Through 14 passages, resistance to the compounds has not been achieved. Most HIV inhibitors with mechanism of antiviral action targeting a viral protein would have selected for a drug resistant virus within 14 passages. These studies indicate that these NCp7-targeted compounds represent new potent anti-HIV drug candidates which could be effectively used in combination with all approved anti-HIV drugs. Active against HIV-1 with EC50's ranging from 0.7 to 16 μM in PBMC and monocytes. Active against drug resistant HIV-1 with EC50's similar to wild type virus. Mechanism of action studies indicate MDH-1-38 is a virucidal agent. Combinations with HIV drugs yielded additivity to synergy with no cytotoxicity. Drug resistance has not been achieved after 14 passage and results in defective virus.
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影响因子：
14.8
作者：
Jenkins, Lisa M. Miller;Ott, David E.;Hayashi, Ryo;Coren, Lori V.;Wang, Deyun;Xu, Qun;Schito, Marco L.;Inman, John K.;Appella, Daniel H.;Appella, Ettore
通讯作者：
Appella, Ettore
影响因子：
--
作者：
Mori M;Kovalenko L;Lyonnais S;Antaki D;Torbett BE;Botta M;Mirambeau G;Mély Y
通讯作者：
Mély Y
影响因子：
4.9
作者：
Buckheit, Robert W., Jr.;Hartman, Tracy L.;Cho, Eui-Hwan
通讯作者：
Cho, Eui-Hwan
影响因子：
3.7
作者：
Dawson, L;Yu, XF
通讯作者：
Yu, XF
影响因子：
56.9
作者：
BERG, JM
通讯作者：
BERG, JM