Protease inhibitors targeting coronavirus and filovirus entry.
Protease inhibitors targeting coronavirus and filovirus entry.
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DOI：
10.1016/j.antiviral.2015.01.011
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发表时间：
2015-04
影响因子：
7.6
通讯作者：
Simmons G
中科院分区：
文献类型：
--
作者：
Zhou Y;Vedantham P;Lu K;Agudelo J;Carrion R Jr;Nunneley JW;Barnard D;Pöhlmann S;McKerrow JH;Renslo AR;Simmons G
我们确定乙烯砜作为埃博拉病毒和SARS冠状病毒的主要候选抑制剂。K11777在亚纳摩尔范围内抑制埃博拉病毒和SARS-CoV进入。强效抑制与P3处存在碱性哌嗪环相关。丝氨酸蛋白酶抑制剂和K11777阻断冠状病毒进入caco-2细胞。卡莫司他保护了10只小鼠中的6只免受SARS-CoV致死感染。为了进入细胞，不同的病毒，包括埃博拉病毒，SARS冠状病毒和新兴的MERS冠状病毒，依赖于宿主细胞蛋白酶对其包膜糖蛋白的激活。因此，相应的酶是抗病毒干预的极好靶标。在细胞培养中，埃博拉病毒以及SARS和MERS冠状病毒的活化可以通过内体半胱氨酸蛋白酶、组织蛋白酶L（CTSL）和组织蛋白酶B（CTSB）来完成。此外，SARS和MERS冠状病毒可以使用位于细胞表面的丝氨酸蛋白酶来激活它们。然而，目前尚不清楚哪些蛋白酶促进病毒在感染宿主中的传播。我们在这里报告，半胱氨酸蛋白酶抑制剂K11777，（（2S）-N-[（1 E，3S）-1-（苯磺酰基）-5-苯基-1-烯-3-基]-2-{[（E）-4-甲基哌嗪-1-羰基]氨基}-3-苯基丙酰胺）和密切相关的乙烯基砜作为广谱抗病毒药物靶向组织蛋白酶介导的细胞进入。K11777已经处于许多寄生虫病（如恰加斯病）的高级开发阶段，并已在一系列动物模型中证明安全有效。K11777对SARS-CoV和埃博拉病毒进入的抑制作用在亚纳摩尔范围内。为了评估半胱氨酸或丝氨酸蛋白酶是否促进病毒在宿主中的传播，我们比较了优化的K11777衍生物与卡莫司他（一种TMPRSS 2和相关丝氨酸蛋白酶的抑制剂）的抗病毒活性。采用SARS-CoV感染的致病性动物模型，我们证明了SARS-CoV的病毒传播和发病机制是由丝氨酸蛋白酶而不是半胱氨酸蛋白酶驱动的，并且可以通过卡莫司他有效地预防。卡莫司他已在临床上用于治疗慢性胰腺炎，因此代表了呼吸道冠状病毒感染的令人兴奋的潜在治疗方法。我们的研究结果表明，卡莫司他或类似的丝氨酸蛋白酶抑制剂可能是治疗SARS和潜在的MERS的有效选择，而乙烯基砜基抑制剂是埃博拉病毒治疗的优秀候选药物。
We identify vinylsulfones as lead candidate inhibitors of Ebola virus and SARS-CoV. K11777 inhibited Ebola virus and SARS-CoV entry in the sub-nanomolar range. Potent inhibition correlated with the presence of a basic piperazine ring at P3. Serine protease inhibitor and K11777 blocked coronavirus entry into caco-2 cells. Camostat protected 6 out of ten mice from lethal infection with SARS-CoV. In order to gain entry into cells, diverse viruses, including Ebola virus, SARS-coronavirus and the emerging MERS-coronavirus, depend on activation of their envelope glycoproteins by host cell proteases. The respective enzymes are thus excellent targets for antiviral intervention. In cell culture, activation of Ebola virus, as well as SARS- and MERS-coronavirus can be accomplished by the endosomal cysteine proteases, cathepsin L (CTSL) and cathepsin B (CTSB). In addition, SARS- and MERS-coronavirus can use serine proteases localized at the cell surface, for their activation. However, it is currently unclear which protease(s) facilitate viral spread in the infected host. We report here that the cysteine protease inhibitor K11777, ((2S)-N-[(1E,3S)-1-(benzenesulfonyl)-5-phenylpent-1-en-3-yl]-2-{[(E)-4-methylpiperazine-1-carbonyl]amino}-3-phenylpropanamide) and closely-related vinylsulfones act as broad-spectrum antivirals by targeting cathepsin-mediated cell entry. K11777 is already in advanced stages of development for a number of parasitic diseases, such as Chagas disease, and has proven to be safe and effective in a range of animal models. K11777 inhibition of SARS-CoV and Ebola virus entry was observed in the sub-nanomolar range. In order to assess whether cysteine or serine proteases promote viral spread in the host, we compared the antiviral activity of an optimized K11777-derivative with that of camostat, an inhibitor of TMPRSS2 and related serine proteases. Employing a pathogenic animal model of SARS-CoV infection, we demonstrated that viral spread and pathogenesis of SARS-CoV is driven by serine rather than cysteine proteases and can be effectively prevented by camostat. Camostat has been clinically used to treat chronic pancreatitis, and thus represents an exciting potential therapeutic for respiratory coronavirus infections. Our results indicate that camostat, or similar serine protease inhibitors, might be an effective option for treatment of SARS and potentially MERS, while vinyl sulfone-based inhibitors are excellent lead candidates for Ebola virus therapeutics.