Antiviral research最新文献_第4页

Inhibition of influenza virus replication by artificial proteins (αReps) targeting its RNA-polymerase 靶向流感病毒rna聚合酶的人工蛋白（α - reps）对流感病毒复制的抑制作用

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-10-21 DOI: 10.1016/j.antiviral.2025.106300

Mélissa Bessonne , Jessica Morel , Quentin Nevers , Julie Groutsch , Agathe Urvoas , Marie Valerio-Lepiniec , Thibaut Crépin , Philippe Minard , Bernard Delmas

Seasonal epidemics and pandemics caused by influenza A viruses still represent a main public health burden in the world. Influenza viruses replicate and transcribe their genome in the nucleus of the infected cells, two functions that are supported by the viral RNA-dependent RNA-polymerase (FluPol) through extensive structural rearrangements and differential interactions with host cell factors. To get insights into its functioning, we screened a phage-display library of biosynthetic proteins (named αReps and built on a rigid alpha-helicoidal HEAT-like scaffold) against the structurally invariant FluPol core and several flexibly-linked domains of the FluPol PB2 subunit. Several αReps specific of the cap binding domain [CBD], the 627-domain and the NLS domain of PB2 displayed FluPol inhibitory and virus neutralization activities when transiently expressed in the cytosol. Furthermore, intracellular ectopic inducible expression of the αReps C3 and F3 (specific of the CBD and the 627-domain, respectively) in influenza virus permissive cells blocked multiplication of viruses representative of the H1N1, H3N2 and H7N1 subtypes, even when induced at late times post-infection. Bispecific αReps constructs (C3-F3 and F3-C3) display a higher FluPol inhibitory activity than their monomeric counterparts. These results suggest that interfering with FluPol structural rearrangements may represent a promising strategy to block virus multiplication and to design new types of antivirals such as dual binders targeting distant sites on FluPol. Furthermore, we found that the 627-domain constitutes a new possible target for engineering influenza antivirals.

季节性流行病和甲型流感病毒引起的大流行仍然是世界上一个主要的公共卫生负担。流感病毒在感染细胞的细胞核中复制和转录其基因组，这两种功能通过广泛的结构重排和与宿主细胞因子的差异相互作用得到病毒rna依赖rna聚合酶（FluPol）的支持。为了深入了解其功能，我们针对结构不变的FluPol核心和FluPol PB2亚基的几个柔性连接结构域筛选了一个生物合成蛋白的噬菌体展示文库（命名为αReps，建立在刚性的α螺旋状热样支架上）。PB2的帽结合域（CBD）、627结构域和NLS结构域特异性α - rep在细胞质中短暂表达时，表现出FluPol抑制和病毒中和活性。此外，在流感病毒允许细胞中，细胞内异位诱导α - reps C3和F3（分别是CBD和627结构域的特异性）的表达阻断了H1N1、H3N2和H7N1亚型代表病毒的增殖，即使在感染后后期诱导也是如此。双特异性α - reps结构体（C3-F3和F3-C3）比它们的单体对应物表现出更高的FluPol抑制活性。这些结果表明，干扰FluPol结构重排可能是阻断病毒增殖和设计新型抗病毒药物（如针对FluPol上远端位点的双结合物）的一种有前途的策略。此外，我们发现627结构域可能成为工程流感抗病毒药物的新靶点。

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引用次数: 0

Pretreatment viral quasispecies characteristics and evolutionary phases correlate with HBsAg seroconversion in peginterferon-alfa-2a-treated children with HBeAg-positive chronic hepatitis B 聚乙二醇干扰素- α -2a治疗的hbeag阳性慢性乙型肝炎患儿中预处理病毒准种特征和进化阶段与HBsAg血清转化相关

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-10-12 DOI: 10.1016/j.antiviral.2025.106291

Juncheng Yang , Guifeng Yang , Haitang He , Hai Liu , Qunfang Fu , Xiaoting Wu , Ran Meng , Zhuoyue Li , Qian Zhao , Kangxian Luo , Zhihua Liu

Background & aims

To evaluate baseline HBV quasispecies (QS) characteristics as a predictor of HBsAg seroconversion in peginterferon-alfa-2a-treated HBeAg-positive pediatric chronic hepatitis B (CHB).

Methods

25 patients achieved HBsAg seroconversion (SS group) and another 25 did not (SN group) were enrolled, matched for baseline profiles. Full-length HBV genomes were obtained from pretreatment serum via clone sequencing. Twelve untreated carriers (CA group) served as controls.

Results

SN group demonstrated significantly higher genetic distance (GD) and mutation frequency index (MFI) in the preCore/Core (preC/C) gene versus SS and CA groups (p < 0.05). Treatment failure exclusively correlated with G1896A/C1913A/C2078G mutations. Jonckheere-Terpstra test confirmed a CA→SS→SN diversity gradient in preC/C (p < 0.05). Non-invasive fibrosis indices increased across CA→SS→SN (p < 0.05) and correlated positively with QS diversity. Evolutionary analyses revealed divergent haplotype clustering in SN versus star-like radiation patterns in CA/SS. Phylogenetic analysis positioned CA/SS haplotypes at earlier evolutionary phases, while SN haplotypes at a later phase. Notably, haplotypes harboring the three aforementioned mutations were also located at the later phase. Positive selection analysis identified two SN-specific mutations (codon 34[C1913A] and 89[C2078G]). A LASSO-derived predictive model (age, C1913A/C2078G, preC/C MFI) yielded the area under the receiver operating characteristic curve (AUROC) 0.814 (sensitivity 92 %, specificity 68 %).

Conclusions

Lower baseline HBV quasispecies diversity predicts Peginterferon-alfa-2a-induced HBsAg seroconversion in pediatric CHB. The observed diversity continuum, association with fibrosis indices, suggests seroconversion occurs preferentially during earlier hepatitis phases. Pretreatment viral genetic profiling may optimize therapeutic timing, highlighting its prognostic value for childhood HBV management.

背景与目的：评估基线HBV准种（QS）特征作为聚乙二醇干扰素- α -2a治疗的hbeag阳性儿童慢性乙型肝炎（CHB） HBsAg血清转化的预测因子。方法：25例患者实现HBsAg血清转化（SS组），另外25例患者未实现HBsAg血清转化（SN组），基线资料匹配。通过克隆测序从预处理血清中获得HBV全长基因组。12例未经治疗的携带者（CA组）作为对照组。结果：与SS和CA组相比，SN组的preore /Core （preC/C）基因遗传距离（GD）和突变频率指数（MFI）显著高于SS和CA组(结论：较低的基线HBV准种多样性预测聚干扰素- α -2a诱导的儿童CHB HBsAg血清转化。观察到的多样性连续体与纤维化指数相关，表明血清转化优先发生在早期肝炎阶段。预处理病毒遗传谱分析可以优化治疗时机，突出其对儿童HBV管理的预后价值。

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引用次数: 0

Antiviral susceptibility monitoring: testing algorithm, methods, and findings for influenza season, 2023–2024 2023-2024年流感季节抗病毒药物敏感性监测：检测算法、方法和结果

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-10-11 DOI: 10.1016/j.antiviral.2025.106299

Mira C. Patel , Ha T. Nguyen , Vasiliy P. Mishin , Philippe Noriel Q. Pascua , Chloe Champion , Mercedes Lopez-Esteva , Angiezel Merced-Morales , Alicia Budd , Marie K. Kirby , Benjamin Rambo-Martin , Jennifer Laplante , Allen Bateman , Kirsten St. George , Maureen Sullivan , John Steel , Rebecca J. Kondor , Larisa V. Gubareva

Antiviral susceptibility monitoring is integral to influenza surveillance conducted by CDC in collaboration with partners. Here, we outlined the algorithm and methods used for assessing antiviral susceptibility of viruses collected during 2023–2024 season. Virus specimens were provided by public health laboratories in the United States (US) and by laboratories in other countries that belong to the Pan American Health Organization. In the US, antiviral susceptibility surveillance conducted nationally is strengthened by sequence-only analysis of additional viruses collected at a state level. Viral genome sequence analysis was the primary approach to assess susceptibility to M2 blockers (n = 5123), neuraminidase (NA) inhibitors (n = 6874), and a polymerase acidic protein (PA) inhibitor (baloxavir, n = 6567). Over 99 % of type A viruses had M2-S31N that confers resistance to M2 blockers. Although oseltamivir-resistant viruses carrying NA-H275Y (N1 numbering) were rare (0.35 %), a cluster of four such viruses was identified in Haiti. Viruses with other NA mutations conferring reduced inhibition by NA inhibitor(s) were also detected sporadically. This includes a cluster of three influenza B viruses in Texas that shared a new mutation, NA-A245G conferring reduced inhibition by peramivir. Three viruses with reduced baloxavir susceptibility were identified, which had PA-I38T, PA-Y24C or PA-V122A; the latter two new mutations identified through augmented approach to sequence analysis. To monitor baseline susceptibility, supplementary in vitro testing was conducted on approximately 7 % of viruses using NA inhibition assay and cell culture-based assay IRINA. Implementation of Sequence First approach provided comprehensive and high throughput methodology for antiviral susceptibility assessment and reduced redundant phenotypic testing.

抗病毒药物敏感性监测是疾病预防控制中心与合作伙伴合作开展的流感监测的组成部分。在此，我们概述了用于评估2023-2024季节收集的病毒抗病毒敏感性的算法和方法。病毒标本由美国的公共卫生实验室和泛美卫生组织其他国家的实验室提供。在美国，通过对州一级收集的其他病毒进行仅序列分析，加强了在全国范围内进行的抗病毒药物敏感性监测。病毒基因组序列分析是评估M2阻滞剂（n=5123）、神经氨酸酶（NA）抑制剂（n=6874）和聚合酶酸性蛋白（PA）抑制剂（baloxavir, n=6567）易感性的主要方法。超过99%的A型病毒含有M2- s31n，这种病毒对M2阻滞剂有抵抗力。虽然携带NA-H275Y （N1编号）的奥司他韦耐药病毒很少见（0.35%），但在海地发现了四种此类病毒。具有其他NA突变的病毒对NA抑制剂的抑制作用降低，也偶有发现。这包括德克萨斯州的三种乙型流感病毒群，它们共有一种新的突变，NA-A245G，使帕拉米韦的抑制作用降低。鉴定出3种降低巴洛韦敏感性的病毒，分别为PA-I38T、PA-Y24C和PA-V122A；后两种新突变是通过扩增序列分析方法鉴定出来的。为了监测基线敏感性，对大约7%的病毒进行了补充体外测试，使用NA抑制试验和基于细胞培养的IRINA试验。序列优先方法的实施为抗病毒药物敏感性评估提供了全面和高通量的方法，减少了冗余的表型检测。

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引用次数: 0

Mapping the impact of 1′-, 2′- and 4′-nucleotide modifications on the Respiratory Syncytial Virus RNA-dependent RNA polymerase 绘制1'-、2'-和4'-核苷酸修饰对呼吸道合胞病毒RNA依赖RNA聚合酶的影响

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-10-11 DOI: 10.1016/j.antiviral.2025.106298

Priscila Sutto-Ortiz , Barbara Selisko , François Ferron , Jean-Pierre Sommadossi , Adel Moussa , Steven Good , Bruno Canard , Etienne Decroly

Human respiratory syncytial virus (RSV) causes pediatric bronchiolitis and severe respiratory illness in the elderly. Despite recent advancements in vaccines and antibody therapies, the search for antiviral agents remains a significant public health challenge. We designed nucleotide analogs (NAs) with ribose modifications to assess their incorporation by the RSV polymerase. Biochemical assays and structural modelling revealed that these NAs effectively disrupt RNA synthesis elongation. They act as chain-terminators via a unique mechanism mediated by the 4′-modification, whereas 2′-F alone has no effect and 1′-modification slows-down RNA synthesis. We evaluated the ability of the polymerase to discriminate between natural nucleotides and NAs through incorporation efficiency/competitive assays, correlating these findings with RSV replication inhibition in infected cell cultures. Our ranking of compounds indicates that cytidine analogs demonstrate the strongest antiviral activity, due to their phosphorylation efficiency and intracellular concentration relative to natural nucleotides as well as their ready incorporation into the growing RNA chain. 4′-modifications are accepted by the RSV polymerase due to structural differences between the active sites of (+) and (−) RNA virus polymerases.

人呼吸道合胞病毒（RSV）引起小儿细支气管炎和老年人严重呼吸道疾病。尽管最近在疫苗和抗体疗法方面取得了进展，但寻找抗病毒药物仍然是一项重大的公共卫生挑战。我们设计了带有核糖修饰的核苷酸类似物（NAs）来评估它们与RSV聚合酶的结合。生化分析和结构建模显示，这些NAs有效地破坏RNA合成延伸。它们通过一种由4'-修饰介导的独特机制作为链终止子，而2'-F单独不起作用，1'-修饰会减慢RNA合成。我们通过整合效率/竞争试验评估了聚合酶区分天然核苷酸和NAs的能力，并将这些发现与感染细胞培养中RSV复制抑制联系起来。我们对化合物的排序表明，胞苷类似物表现出最强的抗病毒活性，这是由于它们的磷酸化效率和相对于天然核苷酸的细胞内浓度，以及它们随时融入生长的RNA链。由于（+）和（-）RNA病毒聚合酶活性位点的结构差异，RSV聚合酶接受4'-修饰。

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引用次数: 0

Small molecule HBV RNA destabilizing drugs: Drugs of the future or compounds from the past? 破坏HBV RNA稳定的小分子药物：未来的药物还是过去的化合物？

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-10-09 DOI: 10.1016/j.antiviral.2025.106288

Timothy M. Block, Dimitar Gotchev, Yanming Du

Small-molecule HBV RNA destabilizing agents, such as the dihydroquinolizinones (DHQs), were first disclosed in a patent filing in 2015 and in peer reviewed literature in 2018. These compounds inhibit Poly-adenylating Polymerases 5 and 7 (PAPD5/7) and represent a novel antiviral strategy and their ability to degrade hepatitis B surface antigen (HBsAg) in cell culture and animal models generated considerable excitement and commercial interest. However, extrahepatic toxicity observed in preclinical and Phase I studies led to the discontinuation of several development programs. The subsequent emergence of liver-targeted PAPD5/7 inhibitors with improved safety profiles has rekindled interest in this therapeutic approach. Yet, with the apparent success of other investigational antivirals in reducing HBsAg levels, such as siRNAs, antisense oligonucleotides, and in at least one example, capsid assembly modulators (CAMs), questions remain as to whether RNA destabilizers still have a role in managing chronic hepatitis B (CHB). This review describes the current status of PAPD5/7 inhibitor development, evaluates the advantages and limitations of the approach, and considers potential strategies for integrating this class of molecules with other HBV therapies.

小分子HBV RNA不稳定剂，如二氢喹啉酮（dhq），于2015年在专利申请中首次披露，2018年在同行评审文献中首次披露。这些化合物抑制聚腺苷酸聚合酶5和7 (PAPD5/7)，代表了一种新的抗病毒策略，它们在细胞培养和动物模型中降解乙型肝炎表面抗原（HBsAg）的能力引起了相当大的兴奋和商业兴趣。然而，在临床前和I期研究中观察到的肝外毒性导致了几个开发项目的中断。随后出现的肝靶向PAPD5/7抑制剂具有更好的安全性，重新引起了人们对这种治疗方法的兴趣。然而，随着其他研究抗病毒药物在降低HBsAg水平方面的明显成功，如sirna，反义寡核苷酸，以及至少一个例子，衣壳组装调节剂（CAMs）， RNA不稳定剂是否仍然在慢性乙型肝炎（CHB）的治疗中发挥作用仍然存在疑问。这篇综述描述了PAPD5/7抑制剂的发展现状，评估了该方法的优势和局限性，并考虑了将这类分子与其他HBV治疗相结合的潜在策略。

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引用次数: 0

Ciclopirox suppresses poxvirus replication by targeting iron metabolism 环匹罗通过靶向铁代谢抑制痘病毒复制。

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-10-09 DOI: 10.1016/j.antiviral.2025.106290

Anil Pant , Djamal Brahim Belhaouari , Lara Dsouza , D.M. Nirosh Udayanga , Zhengqiang Wang , Zhilong Yang

Poxviruses remain a significant global health concern, necessitating the development of novel antiviral strategies. Through high-throughput screening, we previously identified ciclopirox (CPX), an FDA-approved antifungal, as a hit that inhibits vaccinia virus (VACV) replication. Here, we further characterized its antiviral activity and mechanism of action using human primary fibroblasts. CPX significantly reduced VACV titers without reducing host cell viability, with an EC₅₀ in the sub-micromolar range and a CC₅₀ > 500 μM. Rescue experiments demonstrated that CPX inhibits viral replication primarily through chelation of intracellular Fe³⁺ and, to a lesser extent, Fe²⁺, as evidenced by partial restoration of viral replication with ferric ammonium citrate supplementation. Furthermore, overexpression of the iron-dependent enzymes RRM2 and the VACV-encoded F4L reduced the inhibitory effect of CPX, indicating that these host and viral proteins are affected by CPX treatment. Moreover, CPX treatment suppressed cowpox virus and monkeypox (mpox) virus replication in vitro. It also reduced VACV titers in ex vivo mouse lung tissue. These findings highlight host iron metabolism as a critical determinant of poxvirus replication and identify CPX as a promising antiviral candidate against multiple orthopoxviruses.

痘病毒仍然是一个重大的全球健康问题，需要开发新的抗病毒策略。通过高通量筛选，我们先前确定了环匹罗（CPX），一种fda批准的抗真菌药物，作为抑制痘苗病毒（VACV）复制的打击。本研究利用人原代成纤维细胞进一步表征了其抗病毒活性和作用机制。CPX在不降低宿主细胞活力的情况下显著降低了VACV滴度，EC50在亚微摩尔范围内，CC50在500 μM范围内。修复实验表明，CPX主要通过螯合细胞内Fe3+抑制病毒复制，在较小程度上，通过补充柠檬酸铁铵可以部分恢复病毒复制。此外，铁依赖性酶RRM2和vacv编码的F4L的过表达降低了CPX的抑制作用，表明这些宿主和病毒蛋白受到CPX处理的影响。此外，CPX处理能抑制牛痘病毒和猴痘病毒的体外复制。它还降低了离体小鼠肺组织中的VACV滴度。这些发现强调宿主铁代谢是痘病毒复制的关键决定因素，并确定CPX是一种有希望的抗多种痘病毒的抗病毒候选药物。

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引用次数: 0

Deciphering letermovir's mode of action and resistance mutation effects Letermovir的作用方式和抗性突变效应的解读。

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-10-09 DOI: 10.1016/j.antiviral.2025.106289

C. Gourin , T. Flores , C. Lefèvre , S. Alain , G. Ligat , S. Hantz

Human cytomegalovirus is an opportunistic pathogen responsible for severe infections in immunocompromised patients, the leading cause of congenital infections worldwide, and potentially implicated in carcinogenesis. The HCMV terminase complex (pUL56-pUL89-pUL51) has emerged as a key target for antiviral drug development. Letermovir, an antiviral agent targeting this complex, inhibits viral DNA packaging, but resistance-associated mutations have already been identified within subunits. Moreover, the precise mechanism of action of letermovir remains incompletely understood.

We investigated interactions among terminase subunits in presence or absence of letermovir. Wild-type and mutant forms of these proteins (including resistance mutations V236M, L241P, L257I, C325Y, R369M in pUL56 and A95V in pUL51) were cloned into NanoBiT® PPI and pCI-neo vectors. Letermovir was added after transfection in HEK293T cells, and protein-protein interactions were assessed.

Our results show letermovir does not disrupt interactions between wild-type terminase subunits. Resistance-associated mutations modulate the strength of these interactions, with certain mutations (such as pUL56 V236M and L257I) significantly enhancing or reducing binding. Notably, double mutants exhibited synergistic effects. Structural analysis using the AlphaFold3 platform revealed differences between the mutation site of pUL56 and its HSV-1 counterpart pUL28. A hypothetical 3D analysis based on the cryo-EM structure of the HSV-1 terminase complex showed that resistance mutations were oriented outside the complex.

These findings suggest letermovir does not act by directly inhibiting interactions among HCMV terminase subunits. Analysis of resistance-associated mutations provides insight into the molecular basis of HCMV resistance to letermovir and may inform development of novel antiviral strategies targeting the terminase complex.

人类巨细胞病毒是一种机会性病原体，可导致免疫功能低下患者的严重感染，是全世界先天性感染的主要原因，并可能与致癌有关。HCMV末端酶复合物（pUL56-pUL89-pUL51）已成为抗病毒药物开发的关键靶点。Letermovir是一种靶向这种复合体的抗病毒药物，可以抑制病毒DNA包装，但在亚基中已经发现了与耐药性相关的突变。此外，letermovir的确切作用机制尚不完全清楚。我们研究了终端酶亚基之间的相互作用，在存在或不存在的letermovir。这些蛋白的野生型和突变型（包括pUL56中的抗性突变V236M、L241P、L257I、C325Y、R369M和pUL51中的A95V）被克隆到NanoBiT®PPI和pCI-neo载体中。转染HEK293T细胞后加入Letermovir，评估蛋白与蛋白的相互作用。我们的研究结果表明，letermovir不会破坏野生型末端酶亚基之间的相互作用。抗性相关突变调节这些相互作用的强度，某些突变（如pUL56 V236M和L257I）显著增强或减少结合。值得注意的是，双突变体表现出协同效应。利用AlphaFold3平台进行的结构分析显示，pUL56的突变位点与其HSV-1对应的pUL28存在差异。基于HSV-1末端酶复合体的低温电镜结构的假设三维分析表明，抗性突变定向在复合体外。这些发现表明，letermovir不是通过直接抑制HCMV端酶亚基之间的相互作用而起作用的。对耐药相关突变的分析提供了对HCMV对letermovir耐药的分子基础的深入了解，并可能为开发针对末端酶复合物的新型抗病毒策略提供信息。

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引用次数: 0

Composition, three-dimensional structure and formation mechanism of the foot and mouth disease virus replication complexes 口蹄疫病毒复制复合体的组成、三维结构及形成机制。

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-10-08 DOI: 10.1016/j.antiviral.2025.106287

Bonan Lv , Xingran Wang , Ying Zhou , Zihan Su , Yidan Sun , Yingying Yang , Yang Lu , Zishu Pan , Xiao-Feng Tang , Chao Shen

Positive-sense RNA virus infections induce vesicle formation within host cells to support RNA replication. By extracting and purifying foot-and-mouth disease virus (FMDV) replication complexes and comparing host cells and replication complexes through targeted lipidomics and proteomics analyses, we found that FMDV enriches host cell proteins and polyunsaturated fatty acids in viral replication complexes (VRCs) to facilitate their formation. On the basis of these findings, we propose a model in which VRCs progress from single-membrane vesicles to multi-membrane vesicles (MMVs) during FMDV replication, a process that requires coordinated contributions of host cell proteins and organelle membranes derived from multiple organelles. Our study showed that, as infection advances, FMDV converts single-membrane vesicles (SMVs) into MMVs, which aggregate to expand the surface area of the replication platform and enhance replication efficiency. These membrane structures function in FMDV replication; the endoplasmic reticulum undergoes curling and folding to support VRC assembly. Additionally, some VRCs possess outward-facing openings that permit material exchange. These findings reveal unexpected similarities between FMDV and distantly related positive-strand RNA viruses, suggesting that shared host cellular pathways are exploited to construct membrane-bound replication factories.

正义RNA病毒感染诱导宿主细胞内形成囊泡以支持RNA复制。我们通过提取和纯化口蹄疫病毒（FMDV）复制复合体，并通过靶向脂质组学和蛋白质组学分析对宿主细胞和复制复合体进行比较，发现FMDV可富集宿主细胞蛋白和病毒复制复合体（VRCs）中的多不饱和脂肪酸，促进其形成。基于这些发现，我们提出了一个模型，在FMDV复制过程中，VRCs从单膜囊泡到多膜囊泡（mmv），这一过程需要宿主细胞蛋白和来自多个细胞器的细胞器膜的协调贡献。我们的研究表明，随着感染的进展，FMDV将单膜囊泡（smv）转化为mmv，它们聚集在一起，扩大了复制平台的表面积，提高了复制效率。这些膜结构在FMDV复制中起作用；内质网经历卷曲和折叠以支持VRC组装。此外，一些虚拟现实中心拥有面向外的开口，允许材料交换。这些发现揭示了FMDV与远亲正链RNA病毒之间意想不到的相似性，表明利用共享宿主细胞途径构建膜结合复制工厂。

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引用次数: 0

Inhibitors of pyrimidine synthesis synergize with N4-hydroxycytidine to diminish influenza virus replication 嘧啶合成抑制剂与n4 -羟基胞苷协同作用减少流感病毒复制。

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-10-03 DOI: 10.1016/j.antiviral.2025.106286

Leon Schrell , David Scheibner , Antje Dickmanns , Kim M. Stegmann , Lukas Mathias Michaely , Annika Graaf-Rau , Philip Beer , Andrew Parker , Sandra Diederich , Anne Balkema-Buschmann , Matthias Dobbelstein

Influenza viruses remain a major threat to both human and animal health, with seasonal outbreaks and the risk of pandemics caused by reassortant strains. Antiviral drugs are needed as a complement to vaccines, but resistance often limits their long-term efficacy. N4-hydroxycytidine (NHC), the active form of Molnupiravir, shows potent activity against influenza A viruses (IAVs) in both cell cultures and animal models, with minimal resistance observed.

Building on prior work in SARS-CoV-2, we investigated whether inhibiting pyrimidine biosynthesis could enhance NHC's antiviral activity against IAVs. The combination of NHC with inhibitors of dihydroorotate dehydrogenase (DHODH) or cytidine triphosphate synthases (CTPS1/2) showed strong synergy. This was evident through reduced cytopathic effects, decreased viral RNA and protein, and a marked absence of infectious virus particles. This synergy was consistent across multiple IAV subtypes, including H1N1, H1N2, H3N2, and H5N1. This synergistic effect was reversed by exogenously supplemented pyrimidine nucleosides, confirming nucleotide depletion as a key mechanism. However, some avian IAVs were less sensitive to the treatment in mammalian cells. The PB2-K627E mutation, affecting the interaction with host factor ANP32, modulated NHC efficacy, implicating viral adaptation in drug responsiveness.

In a ferret model of H5N1 infection, NHC combined with the CTPS inhibitor STP938 reduced clinical symptoms and lung pathology, with NHC mostly driving antiviral activity and STP938 contributing to disease mitigation. These findings indicate that combining NHC with pyrimidine biosynthesis inhibitors enhances antiviral efficacy against IAVs, especially in rapidly replicating viruses, and may broaden the utility of nucleoside analogues in influenza therapy.

流感病毒仍然是对人类和动物健康的主要威胁，存在季节性疫情和由重组毒株引起的大流行风险。需要抗病毒药物作为疫苗的补充，但耐药性往往限制了它们的长期效力。n4 -羟基胞苷（NHC）是Molnupiravir的活性形式，在细胞培养和动物模型中都显示出对甲型流感病毒（iav）的有效活性，观察到的耐药性最小。在前人对SARS-CoV-2的研究基础上，我们研究了抑制嘧啶生物合成是否可以增强NHC对iav的抗病毒活性。NHC与二氢乙酸脱氢酶（DHODH）或胞苷三磷酸合成酶（CTPS1/2）抑制剂联合使用表现出较强的协同作用。这可以通过降低细胞病变效应、减少病毒RNA和蛋白质以及明显缺乏传染性病毒颗粒来证明。这种协同作用在多种IAV亚型中是一致的，包括H1N1、H1N2、H3N2和H5N1。这种协同效应被外源补充的嘧啶核苷逆转，证实核苷酸耗竭是一个关键机制。然而，一些禽类iav对哺乳动物细胞的处理不太敏感。PB2-K627E突变影响了与宿主因子ANP32的相互作用，从而调节了NHC的疗效，暗示病毒适应了药物反应性。在H5N1感染雪貂模型中，NHC与CTPS抑制剂STP938联合可减轻临床症状和肺部病理，NHC主要驱动抗病毒活性，而STP938有助于减轻疾病。这些发现表明，将NHC与嘧啶类生物合成抑制剂联合使用可增强对iav的抗病毒效果，特别是对快速复制的病毒，并可能扩大核苷类似物在流感治疗中的应用。

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引用次数: 0

Design, synthesis, and biological evaluation of novel 6-aminoalkyl- and 7-heteroaryl substituted 7-deazapurine nucleoside analogs against SARS-CoV-2 replication. 新型抗SARS-CoV-2复制的6-氨基烷基和7-杂芳基取代7-去氮杂嘌呤核苷类似物的设计、合成和生物学评价

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-10-01 Epub Date: 2025-08-07 DOI: 10.1016/j.antiviral.2025.106246

Guoqiang Yao, Xue Shi, Hao Jiang, Anna Duan, Jiancun Zhang

Despite the declaration by the WHO on May 5th of 2023 that the COVID-19 epidemic no longer constitutes a Public Health Emergency of International Concern (PHEIC), it continues to pose a significant threat to human health due to ongoing viral mutations. Although multiple vaccines and drugs have received approval for the prevention and treatment of COVID-19, effectively controlling the epidemic continues to be challenging, emphasizing the necessity for the development of new and more effective antivirals. In this work, we report the design and synthesis of a series of novel 7-alkynyl-7-deazapurine nucleoside analogs and some of their corresponding prodrugs as potential inhibitors of SARS-CoV-2 replication. The biological activities were evaluated and a reasonable structure-activity relationship (SAR) was revealed. Among the compounds, compound 54 (EC₅₀ = 0.71 μM) and 30c (EC₅₀ = 0.66 μM) may serve as potential candidates for further development.

尽管世界卫生组织于2023年5月5日宣布COVID-19流行病不再构成国际关注的突发公共卫生事件（PHEIC），但由于病毒持续突变，它继续对人类健康构成重大威胁。尽管已有多种疫苗和药物获得批准用于预防和治疗COVID-19，但有效控制疫情仍然具有挑战性，这强调了开发新的更有效的抗病毒药物的必要性。在这项工作中，我们设计和合成了一系列新的7-烷基基-7-去氮杂嘌呤核苷类似物及其相应的一些前药，作为SARS-CoV-2复制的潜在抑制剂。对其生物活性进行了评价，揭示了合理的构效关系。其中化合物54 （EC50 = 0.71 μM）和30c （EC50 = 0.66 μM）可能是进一步开发的候选化合物。

引用次数: 0