Antiviral research最新文献_第8页

SMAD5 phosphorylation by ALK1 is modulated by the interaction of the spike protein of SARS-CoV-2 and angiotensin-converting enzyme 2 SMAD5被ALK1磷酸化是由SARS-CoV-2刺突蛋白和血管紧张素转换酶2相互作用调控的

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-08-18 DOI: 10.1016/j.antiviral.2025.106261

Yoichi Teratake , Tadashi Okamura , Yukihito Ishizaka

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused more than 7 million worldwide deaths, but the mechanisms underlying its severe clinical outcomes remain elusive. Although attachment of the spike (S) protein of SARS-CoV-2 to angiotensin-converting enzyme 2 (ACE2), a pivotal step for infection, induces inflammation, the intracellular signals triggered by this interaction are unclear. Here, we found that S protein induced phosphorylation of SMAD family member 5 which was mediated by the bone morphologenetic protein (BMP) receptor activin receptor-like kinase 1 (ALK1) and BMP receptor type II. SMAD5 activation depended on the interaction of the cytoplasmic domains of ACE2 and ALK1. Notably, repetitive treatment with S protein increased the expression of immune-response genes, including interferon regulatory factor 1, interleukin-17A, and cysteine-cysteine chemokine motif ligand 20 (CCL20), all of which were blocked by an ALK inhibitor. In contrast, the S protein of human coronavirus NL63, a pathogen of acute respiratory infection with less severity than SARS-CoV-2 that also binds to ACE2, did not activate SMAD5. Because CCL20 recruits multiple immune cells positive for CC receptor 6, a specific receptor for CCL20, and was detected in broncho-alveolar lavage fluids of patients with respiratory failures by SARS-Cov-2, data suggest that SMAD5 activation by S protein augments the early immune response, leading to the severe clinical outcomes of SARS-CoV-2.

严重急性呼吸综合征冠状病毒2 （SARS-CoV-2）已在全球造成700多万人死亡，但其严重临床结果背后的机制仍然难以捉摸。尽管SARS-CoV-2的刺突蛋白(S)附着于血管紧张素转换酶2 (ACE2)（感染的关键步骤）诱导炎症，但这种相互作用引发的细胞内信号尚不清楚。在这里，我们发现S蛋白诱导SMAD家族成员5磷酸化，该磷酸化是由骨形态发生蛋白（BMP）受体激活素受体样激酶1 （ALK1）和BMP受体II型介导的。SMAD5的激活依赖于ACE2和ALK1胞质结构域的相互作用。值得注意的是，S蛋白的重复处理增加了免疫应答基因的表达，包括干扰素调节因子1、白细胞介素17a和半胱氨酸-半胱氨酸趋化因子基序配体20 (CCL20)，所有这些基因都被ALK抑制剂阻断。相比之下，人类冠状病毒NL63（一种急性呼吸道感染的病原体，其严重程度低于SARS-CoV-2，也与ACE2结合）的S蛋白没有激活SMAD5。由于CCL20招募了多个CC受体6 （CCL20的特异性受体）阳性的免疫细胞，并在SARS-Cov-2呼吸衰竭患者的支气管肺泡灌洗液中检测到，数据表明，S蛋白激活SMAD5增强了早期免疫反应，导致SARS-Cov-2的严重临床结果。

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引用次数: 0

Dexamethasone treatment does not alter mortality but reduces pulmonary pathology in Nipah virus-infected Syrian hamsters 地塞米松治疗不会改变尼帕病毒感染的叙利亚仓鼠的死亡率，但会减少肺部病理

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-08-18 DOI: 10.1016/j.antiviral.2025.106263

Kerry Goldin , Bridget Brackney , Tessa Lutterman , Brandi N. Williamson , Manmeet Singh , Christopher Winski , Kathleen Cordova , Meaghan Flagg , Emmie de Wit

Nipah virus (NiV) is an emerging zoonotic pathogen that causes severe respiratory and neurologic disease, and there are currently no licensed vaccines or approved treatments. The acute respiratory disease caused by NiV is associated with severe inflammation, similar to severe COVID-19. Dexamethasone is an affordable and widely available synthetic glucocorticoid, that improved outcomes when administered to patients with severe COVID-19. To determine whether a similar beneficial effect could be achieved during NiV infection, we tested the effect of an anti-inflammatory or immunosuppressive dose of dexamethasone on NiV in the Syrian hamster model. We found that dexamethasone treatment produced the expected hematologic changes in uninfected animals in a dose-dependent manner. In NiV-infected animals, the anti-inflammatory dose of dexamethasone reduced pulmonary pathology, while the immunosuppressive dose had no effect. The anti-inflammatory dose did not increase virus replication in tissues or virus shedding from the respiratory tract, indicating the anti-inflammatory dose of dexamethasone does not result in increased virus replication. Despite reduced lung pathology, dexamethasone treatment did not increase survival after NiV challenge. When dexamethasone treatment was combined with the antiviral remdesivir, dexamethasone negated the increased survival observed in hamsters treated with remdesivir alone. Our study provides critical information on the effect of dexamethasone administration on the outcome of NiV infection and cautions against the use of dexamethasone in combination with other antivirals like remdesivir without preclinical validation.

尼帕病毒是一种新出现的人畜共患病原体，可引起严重的呼吸道和神经系统疾病，目前没有获得许可的疫苗或批准的治疗方法。NiV引起的急性呼吸道疾病与严重的炎症有关，类似于严重的COVID-19。地塞米松是一种价格合理且可广泛获得的合成糖皮质激素，可改善重症COVID-19患者的预后。为了确定在NiV感染期间是否可以达到类似的有益效果，我们在叙利亚仓鼠模型中测试了抗炎或免疫抑制剂量的地塞米松对NiV的影响。我们发现，在未感染的动物中，地塞米松治疗以剂量依赖的方式产生了预期的血液学变化。在niv感染的动物中，地塞米松的抗炎剂量减轻了肺部病理，而免疫抑制剂量没有作用。抗炎剂量没有增加病毒在组织中的复制或病毒从呼吸道脱落，表明抗炎剂量地塞米松不会导致病毒复制增加。尽管肺部病理减少，地塞米松治疗并没有增加NiV攻击后的生存率。当地塞米松治疗与抗病毒药物瑞德西韦联合使用时，地塞米松抵消了单独使用瑞德西韦治疗的仓鼠的存活率增加。我们的研究提供了关于地塞米松给药对NiV感染结果影响的关键信息，并警告在未经临床前验证的情况下，不要将地塞米松与瑞德西韦等其他抗病毒药物联合使用。

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引用次数: 0

Viral determinants of cis- and trans-cleavage by SARS-CoV-2 Nsp3 and an on-off reporter for monitoring intracellular protease activity SARS-CoV-2 Nsp3顺式和反式切割的病毒决定因素和监测细胞内蛋白酶活性的开关报告基因

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-08-18 DOI: 10.1016/j.antiviral.2025.106262

Fuzhi Lei , Jun Shu , Chengjie Xia , Yahan Lei , Yajie Zhang , Yang Zhang , Zhenghong Yuan , Zhigang Yi

Non-structural protein 3 (nsp3), the largest multifunctional membrane-associated protein encoded by coronaviruses, contains a papain-like protease (PLpro) domain that cleaves viral polypeptides at the nsp1/nsp2, nsp2/nsp3 and nsp3/nsp4 junctions, as well as host substrates such as polyubiquitin chains and ISG15. While host substrate recognition by PLpro is well characterized, the processes by which PLpro identifies and interacts with membrane-associated viral substrates remains poorly defined. In this study, we developed full-length nsp3-containg reporter constructs to investigate PLpro-mediated cleavage in a membrane context. Our results show that differential cleavage sensitivity at nsp2/3 nsp3/4 junction in response to protein expression levels: nsp2/3 cleavage occurred even at low expression levels, whereas efficient nsp3/4 cleavage required high protein dosage, suggesting a reliance on trans-cleavage for the latter. Mutagenesis and structural analysis based on the recently resolved nsp3-4 complex structure suggest that membrane constraints on the substrate, along with positional and conformational requirements of the PLpro domain for cis-cleavage, contribute to the trans-cleavage preference at the nsp3/4 site. Membrane localization and specific protein-protein interactions further facilitate this trans-cleavage process. To assess nsp3/4 cleavage in a more physiologically relevant context, we developed a cleavage-induced degron (CID) system that mimics authentic viral polyprotein processing. This system generates a binary signal upon cleavage at the nsp3/4 junction and response to inhibition by the PLpro inhibitor GRL-0617. Our findings reveal that the domain arrangement of nsp3 dictates trans-dependence of the nsp3/4 cleavage, involving membrane association and protein interactions that may facilitate the subsequent double-membrane vesicle assembly.

非结构蛋白3 （nsp3）是冠状病毒编码的最大的多功能膜相关蛋白，含有一个木瓜蛋白酶样结构域（PLpro），可在nsp1/nsp2、nsp2/nsp3和nsp3/nsp4连接处切割病毒多肽，以及多泛素链和ISG15等宿主底物。虽然PLpro对宿主底物的识别已被很好地表征，但PLpro识别膜相关病毒底物并与之相互作用的过程仍不清楚。在这项研究中，我们开发了全长含nsp3的报告基因构建物来研究在膜环境下plpro介导的裂解。我们的研究结果表明，nsp2/3 / nsp3/4连接的不同切割敏感性响应于蛋白质表达水平：nsp2/3切割即使在低表达水平下也能发生，而有效的nsp3/4切割需要高蛋白质剂量，这表明后者依赖于反式切割。基于最近解析的nsp3-4复合物结构的诱变和结构分析表明，膜对底物的约束，以及PLpro结构域对顺式切割的位置和构象要求，导致了nsp3/4位点的反式切割偏好。膜定位和特定的蛋白质相互作用进一步促进了这种反式切割过程。为了评估nsp3/4在生理相关背景下的裂解，我们开发了一个裂解诱导的降解（CID）系统，模拟真实的病毒多蛋白加工。该系统在nsp3/4连接处裂解并响应PLpro抑制剂GRL-0617的抑制时产生二进制信号。我们的研究结果表明，nsp3结构域的排列决定了nsp3/4切割的反依赖性，包括膜结合和蛋白质相互作用，这可能促进随后的双膜囊泡组装。

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引用次数: 0

Optimization of the monoclonal antibody 3E1 through W32I mutation enhances antiviral efficacy against influenza virus subtypes H1N1 and H3N2 通过W32I突变优化单克隆抗体3E1增强抗病毒H1N1和H3N2亚型流感病毒的效果

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-08-15 DOI: 10.1016/j.antiviral.2025.106260

Shanshan Guan , Qingyu Wang , Jiaojiao Nie , Xin Yao , Kunpeng Xie , Weihao Zhao , Yaotian Chang , Lvzhou Zhu , Jiaru Hui , Tieyan Yin , Xiaopan Liu , Yingxin Zhang , Yaming Shan

The influenza virus has caused a global pandemic with significant morbidity and mortality, underscoring the need to optimize antibodies for improved antiviral efficacy. The monoclonal antibody 3E1 effectively neutralizes Group 1 influenza subtypes, H1 and H5, by inhibiting acid-induced conformational changes of hemagglutinin (HA). However, its neutralizing activity is relatively weak against the Group 2 subtype, H3. In order to broaden the neutralizing activity of wild-type 3E1 (3E1-WT) against both Group 1 and Group 2 viral strains, single-point mutants (3E1-L [W32I], 3E1-H [F103I]) and a double mutant (3E1-H + L [F103I, W32I]) were designed and generated in this study. The binding affinity, microneutralizing activity, and antiviral mechanisms of the mutants were evaluated in vitro. Notably, the 3E1-L mutant exhibited significantly enhanced antiviral activity against H1N1 and H3N2 compared to 3E1-WT, inhibiting both viral entry and release. In vivo studies also indicate that 3E1-L significantly enhances both prophylactic and therapeutic efficacy against A/New-York/61/2015-CDC-LV16A (H1N1) and A/Hong Kong/17/2014/8296 (H3N2). Molecular dynamics simulations of the 3E1-L/HA complex revealed that the W32I mutation could reduce steric hindrance between tryptophan at position 32 and the complementarity-determining region L1 loop of HA. In conclusion, the W32I substitution enhances the antiviral activity of 3E1-WT, suggesting that the optimization of 3E1-L a promising strategy for the development of more effective influenza therapy and prevention.

流感病毒已引起全球大流行，发病率和死亡率都很高，因此需要优化抗体以提高抗病毒效果。单克隆抗体3E1通过抑制酸诱导的血凝素（HA）构象变化，有效中和1组流感亚型H1和H5。然而，它对第二组亚型H3的中和活性相对较弱。为了扩大野生型3E1 （3E1- wt）对第1组和第2组病毒株的中和活性，本研究设计并产生了单点突变体3E1-L [W32I]、3E1- h [F103I]和双点突变体3E1- h +L [F103I， W32I]。在体外对突变体的结合亲和力、微中和活性和抗病毒机制进行了评价。值得注意的是，与3E1-WT相比，3E1-L突变体对H1N1和H3N2的抗病毒活性显著增强，抑制了病毒的进入和释放。体内研究也表明，3E1-L对A/New-York/61/2015-CDC-LV16A （H1N1）和A/Hong Kong/17/2014/8296 （H3N2）的预防和治疗效果均有显著增强。3E1-L/HA复合物的分子动力学模拟表明，W32I突变可以降低32位色氨酸与HA互补决定区L1环之间的位阻。综上所述，W32I的替换增强了3E1-WT的抗病毒活性，这表明3E1-L的优化是开发更有效的流感治疗和预防的有希望的策略。

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引用次数: 0

First evidence of efficacy of peptides targeting the pUL56-pUL89 interaction domain of the human cytomegalovirus terminase complex 首个针对人巨细胞病毒终止酶复合物的pUL56-pUL89相互作用结构域的肽的有效性证据。

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-08-14 DOI: 10.1016/j.antiviral.2025.106259

Sarah Mafi , Jean-Luc Poyet , Sophie Alain , Gaëtan Ligat , Sébastien Hantz

Human cytomegalovirus (HCMV) is a major cause of morbidity in immunocompromised patients and the leading viral cause of congenital infection. The toxicity and emergence of resistance associated with current antivirals underscore the need for alternative therapeutic strategies. The viral terminase complex (pUL56-pUL89-pUL51), essential for genome cleavage and packaging and without homologs in mammalian cells, represents a promising antiviral target. This study evaluated the antiviral potential of peptides targeting the pUL56–pUL89 interaction domain. Peptides derived from the minimal interaction domain between pUL56 and pUL89 (WF10: ₆₇₁WMVVKYMGFF₆₈₀) or an extended sequence (PD17: ₆₆₈PSEWMVVKYMGFFNFSD₆₈₄) were synthesized to interfere with this interaction. To optimize intracellular delivery, peptides were conjugated to a cell-penetrating peptide (CPP) derived from either the HIV-1 transactivator of transcription (TAT) or the Antennapedia homeodomain of Drosophila melanogaster (penetratin; PT). Peptide candidates- PT-WF10, TAT-WF10, and PT-PD17- were evaluated in cellular models for cytotoxicity, hemolysis, antiviral activity, and intracellular distribution. TAT-WF10 and PT-PD17 significantly reduced the cytopathic foci in HCMV-infected cells, with IC₅₀ values of 58 μM and 39 μM, respectively. PT-WF10 lacked antiviral activity, induced significant cytotoxicity and hemolysis, and was mainly localized in the cytoplasm, with only minimal nuclear signal. TAT-WF10 showed cytoplasmic and nuclear distribution, no hemolysis, but induced long-term cytotoxicity from 40 μM. PT-PD17 exhibited cytoplasmic and nuclear distribution, with no significant cytotoxicity or hemolysis up to 80 μM. This study provides the first proof of concept that a peptide targeting the pUL56–pUL89 interaction domain can inhibit HCMV replication. PT-PD17 demonstrated antiviral activity, intracellular distribution, and a favorable safety profile.

人巨细胞病毒（HCMV）是免疫功能低下患者发病的主要原因，也是先天性感染的主要病毒原因。与当前抗病毒药物相关的毒性和耐药性的出现强调了寻找替代治疗策略的必要性。病毒末端酶复合体（pUL56-pUL89-pUL51）在哺乳动物细胞中是基因组切割和包装所必需的，没有同源物，是一个很有前途的抗病毒靶点。本研究评估了靶向pUL56-pUL89相互作用结构域的肽的抗病毒潜力。从pUL56和pUL89之间的最小相互作用域（WF10: 671WMVVKYMGFF680）或扩展序列（PD17: 668PSEWMVVKYMGFFNFSD684）衍生的肽被合成来干扰这种相互作用。为了优化细胞内递送，将多肽偶联到细胞穿透肽（CPP）上，CPP来源于HIV-1转录反激活子（TAT）或果蝇触角同源结构域(penetratin；PT)。候选肽- PT-WF10、TAT-WF10和PT-PD17-在细胞模型中评估细胞毒性、溶血、抗病毒活性和细胞内分布。TAT-WF10和PT-PD17显著降低了hcmv感染细胞的病变灶，IC50值分别为58 μM和39 μM。PT-WF10缺乏抗病毒活性，具有明显的细胞毒性和溶血作用，主要存在于细胞质中，只有极少的核信号。TAT-WF10呈细胞质和细胞核分布，无溶血作用，但在40 μM下具有长期细胞毒性。PT-PD17呈细胞质和细胞核分布，在80 μM范围内无明显的细胞毒性和溶血作用。这项研究首次证明了靶向pUL56-pUL89相互作用结构域的肽可以抑制HCMV复制。PT-PD17表现出抗病毒活性、细胞内分布和良好的安全性。

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引用次数: 0

N-myristoyltransferase inhibitors as candidate broad-spectrum antivirals to treat viral infections promoted by immunosuppression associated with JAK inhibitors therapy n -肉豆蔻酰基转移酶抑制剂作为候选广谱抗病毒药物治疗与JAK抑制剂治疗相关的免疫抑制促进的病毒感染。

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-08-11 DOI: 10.1016/j.antiviral.2025.106258

Haydar Witwit, Juan C. de la Torre

The use of Janus kinase (JAK) inhibitors in the clinic has been expanded significantly during the last decade. However, the immunosuppressive effects of JAK inhibitors, via modulation of key innate cellular signaling pathways, can predispose treated patients to infections, and can also result in reduced control of silent infections and increased risk of reactivation of opportunistic infections. Thus, the JAK inhibitor ruxolitinib, approved for the treatment of myelofibrosis and polycythemia vera, has been shown to exerts a proviral activity during infection with different viruses. Therefore, the clinical relevance of developing antiviral treatments that can be effective in the presence of JAK inhibitors. N-terminal myristoyl transferase (NMT) inhibitors have been shown to exhibit potent antiviral activity against different viruses. Here we document that in the presence of ruxolitinib, NMT inhibitors retain their potent antiviral activity against different viruses, including HSV-1. Our findings support that NMT inhibitors should be explored as therapeutics to treat viral infections associated with immunosuppression caused by treatments with JAK inhibitors.

在过去的十年中，Janus激酶（JAK）抑制剂在临床中的应用已经显著扩大。然而，JAK抑制剂的免疫抑制作用，通过调节关键的先天细胞信号通路，可使接受治疗的患者易受感染，也可导致沉默感染的控制减少和机会性感染重新激活的风险增加。因此，被批准用于治疗骨髓纤维化和真性红细胞增多症的JAK抑制剂ruxolitinib已被证明在不同病毒感染期间发挥前病毒活性。因此，开发在存在JAK抑制剂的情况下有效的抗病毒治疗具有临床意义。n端肉豆蔻酰基转移酶（NMT）抑制剂已被证明对不同的病毒具有有效的抗病毒活性。在这里，我们证明在ruxolitinib的存在下，NMT抑制剂保留了对不同病毒（包括HSV-1）的有效抗病毒活性。我们的研究结果支持NMT抑制剂应该作为治疗JAK抑制剂治疗引起的免疫抑制相关病毒感染的治疗方法。

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引用次数: 0

The protease inhibitor Nirmatrelvir synergizes with inhibitors of GRP78 to suppress SARS-CoV-2 replication 蛋白酶抑制剂Nirmatrelvir与GRP78抑制剂协同抑制SARS-CoV-2复制。

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-07-29 DOI: 10.1016/j.antiviral.2025.106247

Doha Al Krad , Kim M. Stegmann , Antje Dickmanns , Priya Kumar , Claudia Blaurock , Björn-Patrick Mohl , Sina Jasmin Wille , Angele Breithaupt , Tobias Britzke , Anne Balkema-Buschmann , Matthias Dobbelstein

Nirmatrelvir, the active compound of the drug Paxlovid, inhibits the Main protease of SARS-CoV-2 (M^Pro, 3CL^Pro, NSP5). Its therapeutic application reduces but does not abolish the progression of COVID-19 in humans. Here we report a strong synergy of Nirmatrelvir with inhibitors of the ER chaperone GRP78 (HSPA5, BiP). Combining Nirmatrelvir with the GRP78-antagonizing drug candidate HA15 strongly inhibits the replication of SARS-CoV-2, to a far greater extent than either drug alone, as observed by diminished cytopathic effect, levels of detectable virus RNA, TCID₅₀ titers, and reduced accumulation of the non-structural proteins, as well as Spike and N proteins. The original SARS-CoV-2 strain as well as an Omicron variant were similarly susceptible towards the drug combination. Other GRP78 inhibitors or siRNAs targeting GRP78 also fortified the antiviral effect of Nirmatrelvir. In a hamster model of COVID-19, the combination of Nirmatrelvir with HA15 alleviated pneumonia-induced pulmonary atelectasis more effectively than the single drugs. In conclusion, inhibition of the virus Main protease and cellular GRP78 cooperatively diminishes virus replication and may improve COVID-19 therapy.

药物Paxlovid的活性化合物Nirmatrelvir抑制SARS-CoV-2的主要蛋白酶（MPro, 3CLPro, NSP5）。它的治疗应用减少但不能消除COVID-19在人类中的进展。在这里，我们报告了Nirmatrelvir与ER伴侣GRP78 （HSPA5, BiP）抑制剂的强协同作用。Nirmatrelvir与grp78拮抗候选药物HA15联合使用对SARS-CoV-2复制的抑制作用远远大于单独使用任何一种药物，这可以通过降低细胞病变效应、检测到的病毒RNA水平、TCID50滴度以及减少非结构蛋白、Spike和N蛋白的积累来观察到。最初的SARS-CoV-2菌株以及一种欧米克隆变体对这种药物组合同样敏感。其他GRP78抑制剂或靶向GRP78的sirna也增强了Nirmatrelvir的抗病毒作用。在新冠肺炎仓鼠模型中，Nirmatrelvir与HA15联合使用比单药更有效地缓解肺炎性肺不张。总之，抑制病毒主蛋白酶和细胞GRP78共同减少病毒复制，可能改善COVID-19治疗。

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引用次数: 0

Discovery of broad-spectrum antivirals targeting viral proteases using in silico structural modeling and cellular analysis 利用硅结构建模和细胞分析发现针对病毒蛋白酶的广谱抗病毒药物

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-07-29 DOI: 10.1016/j.antiviral.2025.106245

Dharmeshkumar Patel , Ramyani De , Niloufar Azadi , Sujin Lee , Savannah Shooter , Sarah Amichai , Shaoman Zhou , Danielle Monroe , Cameron Mahanke , Tamara R. McBrayer , Michael Muczynski , Abdullah Al-Homoudi , Joseph Engel , Yury A. Bochkov , James E. Gern , Ladislau C. Kovari , Franck Amblard , Raymond F. Schinazi

The development of broad-spectrum antivirals is a high-priority goal to prevent future global outbreaks. Some antiviral agents developed for specific viral protein targets may exhibit broad-spectrum antiviral activity or provide helpful information for broad-spectrum drug development. In this study, we compared the sequence- and structure-based similarity of SARS-CoV-2 3CL^pro with proteases from other viruses and identified 24 proteases with similar active-site structures. Our in-house lead molecules, NIP-22c and CIP-1 were reported as novel peptidomimetic, reversible covalent inhibitors of SARS-CoV-2 3CL^pro with nanomolar potency. Molecular docking of NIP-22c, CIP-1 and nirmatrelvir were performed with structurally similar proteases of different viruses, norovirus, enterovirus and rhinovirus. The predictions were validated with in vitro enzymatic and cell-based assays. As predicted, NIP-22c and CIP-1 showed broad-spectrum antiviral activity with EC₅₀ values in the nanomolar range against SARS-CoV-2, norovirus, enterovirus and rhinovirus by targeting 3CL/3C^pro. In contrast, nirmatrelvir did not show activity up to 10 μM against all three viruses and the mechanism of inactivity of nirmatrelvir was hypothesized through binding pocket analysis using molecular dynamics simulations.

开发广谱抗病毒药物是预防未来全球疫情的高度优先目标。一些针对特定病毒蛋白靶点开发的抗病毒药物可能表现出广谱抗病毒活性或为广谱药物开发提供有用的信息。在这项研究中，我们比较了SARS-CoV-2 3CLpro与其他病毒蛋白酶的序列和结构相似性，并鉴定出24种具有相似活性位点结构的蛋白酶。据报道，我们的内部铅分子NIP-22c和CIP-1是具有纳米级效力的新型肽类可逆性共价sars - cov - 23clpro抑制剂。将NIP-22c、CIP-1和nirmatrelvir与不同病毒（诺如病毒、肠病毒和鼻病毒）结构相似的蛋白酶进行分子对接。这些预测通过体外酶和细胞实验得到了验证。结果表明，NIP-22c和CIP-1对SARS-CoV-2、诺如病毒、肠道病毒和鼻病毒具有广谱抗病毒活性，EC50值在纳摩尔范围内。相比之下，nirmatrelvir对这三种病毒的活性均不超过10 μM，并通过分子动力学模拟的结合袋分析假设了nirmatrelvir无活性的机制。

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引用次数: 0

Suppression of herpes simplex virus type 1 replication by Pin1 inhibitors: insights from H-77 and novel compounds Pin1抑制剂抑制1型单纯疱疹病毒复制：来自H-77和新化合物的见解

IF 4.5 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-07-25 DOI: 10.1016/j.antiviral.2025.106244

Abeer Mohamed Abdelfattah Elsayed , Miuko Kurose , Akifumi Higashiura , Akima Yamamoto , Toshihito Nomura , Takashi Irie , Masaya Fukushi , Jeffrey Encinas , Hisanaka Ito , Takayoshi Okabe , Tomoichiro Asano , Takemasa Sakaguchi

Herpes simplex virus type 1 (HSV-1) infection is widespread globally, necessitating the development of new therapeutic approaches. Previous studies have demonstrated that peptidyl-prolyl cis/trans isomerase Pin1 is essential for the replication of cytomegalovirus, a member of the herpesvirus family. Our research demonstrated that Pin1 knockdown significantly suppressed HSV-1 replication. Furthermore, we found that our Pin1 inhibitor H-77, along with four novel Pin1 inhibitors, also inhibited HSV-1 replication. The 50 % effective concentration (EC₅₀) of H-77 against HSV-1 replication in VeroE6 cells was 0.75 μM. In HSV-1-infected cells treated with H-77, expression levels of the immediate early viral protein ICP0 and late viral proteins VP5 and glycoprotein C (gC) were significantly reduced, indicating suppression of viral protein expression. Immunofluorescence staining revealed that in H-77-treated cells, viral proteins including VP5 were confined within the nucleus by an intact nuclear lamina. Transmission electron microscopy analysis demonstrated that H-77-treated cells exhibited markedly fewer extracellular viral particles, with nucleocapsid nuclear egress being inhibited. These results demonstrate that H-77 suppresses HSV-1 replication through dual mechanisms: inhibition of viral protein synthesis and blockade of nucleocapsid nuclear egress. These findings indicate that Pin1 represents a promising therapeutic target for HSV-1 inhibition, warranting further development of Pin1 inhibitors as anti-HSV-1 agents.

1型单纯疱疹病毒（HSV-1）感染在全球广泛存在，需要开发新的治疗方法。先前的研究表明，肽基脯氨酸顺/反式异构酶Pin1对巨细胞病毒（疱疹病毒家族的一员）的复制至关重要。我们的研究表明，Pin1敲低可显著抑制HSV-1复制。此外，我们发现我们的Pin1抑制剂H-77以及四种新的Pin1抑制剂也能抑制HSV-1的复制。H-77在VeroE6细胞中抑制HSV-1复制的50%有效浓度（EC50）为0.75 μM。在H-77处理的hsv -1感染细胞中，直接早期病毒蛋白ICP0和晚期病毒蛋白VP5和糖蛋白C （gC）的表达水平显著降低，表明病毒蛋白表达受到抑制。免疫荧光染色显示，在h -77处理的细胞中，包括VP5在内的病毒蛋白被完整的核层限制在细胞核内。透射电镜分析表明，h -77处理的细胞细胞外病毒颗粒明显减少，核衣壳核出口受到抑制。这些结果表明，H-77通过抑制病毒蛋白合成和阻断核衣壳核出口的双重机制抑制HSV-1的复制。这些发现表明，Pin1是抑制HSV-1的一个有希望的治疗靶点，需要进一步开发Pin1抑制剂作为抗HSV-1药物。

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Cerebrospinal fluid concentrations, viral suppression and biomarkers with long-acting cabotegravir and rilpivirine intramuscular injections 长效卡博特重力韦和利匹韦林肌肉注射的脑脊液浓度、病毒抑制和生物标志物。

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-07-23 DOI: 10.1016/j.antiviral.2025.106243

Aylin Yilmaz , Amedeo De Nicolò , Antonio D'Avolio , Magnus Gisslén

Our aim was to determine cerebrospinal fluid (CSF) and plasma concentrations of cabotegravir and rilpivirine (CAB/RPV) when administered as long-acting (LA) intramuscular injections every four (Q4W) or eight (Q8W) weeks, and to study the effect on viral suppression and CSF biomarkers of inflammation and neuronal injury. We included six adults with HIV receiving LA CAB/RPV Q4W or Q8W. CSF and plasma concentrations of CAB/RPV (15 samples) were analyzed by UHPLC-MS/MS. We also measured CSF and plasma HIV RNA, CSF and serum neopterin, CSF and serum β2-microglubulin, IgG index, albumin ratio, and CSF NfL. Median (range) total plasma cabotegravir concentrations were 1375 (963–2422) ng/mL, and in CSF 3.48 (1.47–7.60 ng/mL). For rilpivirine, concentrations were 93 (40–157) ng/mL and 1.21 (0.55–1.67) ng/mL, respectively. All participants hade CSF and plasma HIV RNA <20 copies/mL on every occasion. There were no significant changes in any of the CSF biomarkers in participants after switching to LA CAB/RPV. The combination of LA CAB/RPV can achieve therapeutic CSF concentrations throughout the dosing intervals and may suppress HIV replication in the CNS. No significant changes in CSF biomarkers of inflammation or neuronal injury were observed, indicating a neutral CNS effect.

我们的目的是测定cabotegravir和rilpivirine （CAB/RPV）作为长效（LA）肌肉注射每4周（Q4W）或8周（Q8W）给药时脑脊液（CSF）和血浆浓度，并研究对病毒抑制和脑脊液生物标志物炎症和神经元损伤的影响。我们纳入了6名接受LA CAB/RPV Q4W或Q8W治疗的成年HIV感染者。采用UHPLC-MS/MS对15例患者的脑脊液和血浆CAB/RPV浓度进行分析。同时测定脑脊液和血浆HIV RNA、脑脊液和血清neopterin、脑脊液和血清ß2-微球蛋白、IgG指数、白蛋白比值、脑脊液NfL。中位（范围）血浆cabote重力韦总浓度为1375 (963-2422)ng/mL， CSF中为3.48 （1.47-7.60 ng/mL）。利匹韦林的浓度分别为93 (40-157)ng/mL和1.21 (0.55-1.67)ng/mL。所有参与者每次都有CSF和血浆HIV RNA < 20拷贝/mL。转换到LA CAB/RPV后，参与者的任何CSF生物标志物均无显著变化。LA CAB/RPV联合使用可以在整个给药间隔内达到治疗性CSF浓度，并可能抑制HIV在中枢神经系统中的复制。脑脊液炎症或神经元损伤的生物标志物未见明显变化，表明中性中枢神经系统效应。

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