Antiviral research最新文献_第4页

Poly (allylamine hydrochloride)-selenium nanoparticles inhibit porcine reproductive and respiratory syndrome virus by targeting DDX5 and reactive oxygen species 聚丙烯胺-硒纳米粒子对猪繁殖与呼吸综合征病毒DDX5和活性氧的抑制作用

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2026-01-01 Epub Date: 2025-12-05 DOI: 10.1016/j.antiviral.2025.106324

Wenqiao Yang , Yuan Xu , Runmin Kang , Min Zhang , Yuting Zhang , Zeng Yang , Huiyuan Jing , Yilin Liu , Jiaxiang Li , Yongjie Ge , Haibo Feng , Long Zhou , Jiang Pi , Jie Liu

Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) remains a critical threat to the swine industry worldwide due to its rapid evolution and limited vaccine efficacy. In this study, we synthesized and characterized poly (allylamine hydrochloride)-selenium nanoparticles (PAH-Se NPs) and evaluated their antiviral activity against PRRSV in vitro and the underlying mechanism. PAH-Se NPs exhibited uniform spherical morphology with average hydrodynamic diameter of 118 nm and excellent colloidal stability with a zeta potential of +50.7 mV. Cytotoxicity assays demonstrated low toxicity of PAH-Se NPs in MARC-145 cells with a CC₅₀ of 40.24 μg/mL, indicating favorable biocompatibility. Multiple complementary assays, including viral RNA quantification, cell viability restoration, plaque assays, N protein expression analyses, and indirect immunofluorescence assay, showed that PAH-Se NPs could significantly inhibit PRRSV replication in a dose-dependent manner, with an IC₅₀ of 0.076 μg/mL. Moreover, this inhibition was equally effective against other sub-genotypes of PRRSV, including VR2332 and NADC30-like variants, as well as the DNA virus and bacteria, indicating the broad-spectrum anti-microbial properties of PAH-Se NPs. Mechanistic studies indicated that PAH-Se NPs target viral invasion and replication by down-regulating the host factor DDX5, which interacts with PRRSV Nsp9 to facilitate viral RNA synthesis, and suppressed PRRSV-induced reactive oxygen species (ROS) accumulation. This work emphasized PAH-Se NPs as a potent, low-toxicity antiviral candidate, offering promising potential for the development of alternative therapeutics against PRRSV and related viral infections.

猪繁殖与呼吸综合征病毒（PRRSV）由于其快速演变和有限的疫苗效力，仍然是全球养猪业的严重威胁。本研究合成并表征了聚丙烯胺-硒纳米粒子（PAH-Se NPs），并对其体外抗PRRSV的活性及机制进行了研究。PAH-Se NPs具有均匀的球形形貌，平均水动力直径为118 nm，具有良好的胶体稳定性，zeta电位为+50.7 mV。细胞毒性实验表明，PAH-Se NPs对MARC-145细胞的毒性较低，CC50为40.24 μg/mL，具有良好的生物相容性。包括病毒RNA定量、细胞活力恢复、斑块分析、N蛋白表达分析和间接免疫荧光分析在内的多项互补分析表明，PAH-Se NPs能够以剂量依赖性方式显著抑制PRRSV的复制，IC50为0.076 μg/mL。此外，这种抑制对PRRSV的其他亚基因型（包括VR2332和nadc30样变体）以及DNA病毒和细菌同样有效，表明PAH-Se NPs具有广谱抗微生物特性。机制研究表明，PAH-Se NPs通过下调与PRRSV Nsp9相互作用促进病毒RNA合成的宿主因子DDX5，抑制PRRSV诱导的活性氧（ROS）积累，从而靶向病毒的入侵和复制。这项工作强调了PAH-Se NPs是一种有效的、低毒的抗病毒候选药物，为开发抗PRRSV和相关病毒感染的替代疗法提供了有希望的潜力。

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引用次数: 0

Update on combination therapies against HBV in clinical investigations 针对HBV的联合治疗在临床研究中的最新进展。

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2026-01-01 Epub Date: 2025-11-29 DOI: 10.1016/j.antiviral.2025.106321

Lung-Yi Mak , Anna SF. Lok

Functional cure has been proposed to be the treatment endpoint of cure therapies in chronic hepatitis B (CHB), yet it is rarely achieved with monotherapy of novel virus-targeting agents or immunomodulators. Although translation inhibitors – small interfering RNAs (siRNAs) and antisense oligonucleotides can produce marked decline in hepatitis B surface antigen (HBsAg) levels, the response is often not sustained and HBsAg seroclearance rarely occur after treatment cessation, suggesting that pharmacological reduction in HBsAg level may be insufficient in restoring HBV-specific immune response. Increasing number of studies have adopted the combination approach with virus-directing agent(s) plus immunomodulator(s). To date, the most effective regimen involves the concurrent or sequential use of siRNA with PEG-IFNα for 48 weeks, with resultant off-therapy HBsAg seroclearance rates approaching 30 %, and functional cure rates of up to 10 %. Other immunomodulators studied in combination with siRNA such as toll-like receptor agonists, therapeutic vaccines, monoclonal hepatitis B surface antibodies, and immune checkpoint inhibitors are less effective. Almost all studies included NUC and only a few evaluated protocolized NUC withdrawal; thus, few studies have truly evaluated functional cure. Low baseline HBsAg level is the most reliable predictor of HBsAg seroclearance, with many trials exclusively enrolling patients with HBsAg level <200 to <1000 IU/mL. While recent studies have shown promise, further research is needed to determine the optimal classes of drugs to combine, duration of use for each drug and whether they should be used concurrently or sequentially, to meet the desired goal of 30 % functional cure rate.

功能性治愈已被认为是慢性乙型肝炎（CHB）治愈治疗的终点，但仅靠新型病毒靶向药物或免疫调节剂的单一治疗很少能实现。虽然翻译抑制剂-小干扰rna （sirna）和反义寡核苷酸可以使乙型肝炎表面抗原（HBsAg）水平显著下降，但这种反应往往不能持续，停止治疗后很少出现HBsAg血清清除，提示HBsAg水平的药理学降低可能不足以恢复hbv特异性免疫反应。越来越多的研究采用病毒导向剂加免疫调节剂的联合方法。迄今为止，最有效的方案包括siRNA与PEG-IFNα同时或顺序使用48周，由此产生的HBsAg停药血清清除率接近30%，功能治愈率高达10%。与siRNA联合研究的其他免疫调节剂，如toll样受体激动剂、治疗性疫苗、单克隆乙型肝炎表面抗体和免疫检查点抑制剂的效果较差。几乎所有的研究都包括NUC，只有少数研究评估了NUC的退出；因此，很少有研究真正评估功能性治愈。低基线HBsAg水平是HBsAg血清清除率最可靠的预测指标，许多试验只招募HBsAg水平的患者

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引用次数: 0

Suppression of HBV replication and expression by CRISPR/Cas9 ribonucleoproteins CRISPR/Cas9核糖核蛋白抑制HBV复制和表达

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2026-01-01 Epub Date: 2025-12-09 DOI: 10.1016/j.antiviral.2025.106326

Addison C. Hill , Madison B. Schank , Yi Zhang , Ning Sun , Ling Wang , Juan Zhao , Puja Banik , Jaeden S. Pyburn , Holly Orfield , Janet W. Lightner , Tabitha O. Leshaodo , Xiao Y. Wu , Shunbin Ning , Mohamed El Gazzar , Jonathan P. Moorman , Haitao Guo , Zhi Q. Yao

HBV infection is a global public health problem. The current treatment using nucleotide analogues (NA) can suppress viral replication but cannot eliminate HBV infection due to the persistence of covalently closed circular DNA (cccDNA), which sustains HBV replication and integration into the host cell genome and is refractory to NA treatment. CRISPR/Cas9 has been used to disrupt integrated HBV DNA and minichromosomal cccDNA for HBV suppression, but its expression and delivery require viral or non-viral vectors, which pose safety concerns for human application. We have previously reported the use of synthetic guide RNA (gRNA)/Cas9 ribonucleoprotein (RNP) as a non-viral formulation for HBV gene editing and viral suppression. To formulate highly effective CRISPR/Cas9 modalities for HBV gene therapy, here we designed additional gRNA/Cas9 RNPs and compared their antiviral efficacy in HBV-transfected as well as -infected cells. We found that two selected gRNA/Cas9 RNPs (gRNA5/Cas9, gRNA9/Cas9, and particularly their combinations) elicited the most potent antiviral efficacy, as evidenced by the significant inhibition of HBV DNA, RNA, and protein productions. DNA sequencing of the treated cells revealed moderate to high rates of insertion and deletion (indel) or knock-out (KO) mutations at the HBV target genes. Gene alignment analysis showed a high level of conservation for both gRNA5 and gRNA9 target sequences across major HBV genotypes, indicating that these CRISPR-based gene editing therapeutics have the potential to target different HBV strains worldwide. Thus, these synthetic gRNA/Cas9 RNPs represent promising novel therapeutics that can be developed and utilized for HBV gene disruption and viral eradication.

乙型肝炎病毒感染是一个全球性的公共卫生问题。目前使用核苷酸类似物（NA）的治疗可以抑制病毒复制，但由于共价闭合环状DNA （cccDNA）的持续存在，无法消除HBV感染，cccDNA维持HBV复制并整合到宿主细胞基因组中，并且NA治疗难治性。CRISPR/Cas9已被用于破坏整合的HBV DNA和小染色体cccDNA以抑制HBV，但其表达和递送需要病毒或非病毒载体，这对人类应用存在安全性问题。我们之前报道过使用合成向导RNA (gRNA)/Cas9核糖核蛋白（RNP）作为HBV基因编辑和病毒抑制的非病毒制剂。为了制定高效的CRISPR/Cas9模式用于HBV基因治疗，我们设计了额外的gRNA/Cas9 RNPs，并比较了它们在HBV转染和-感染细胞中的抗病毒效果。我们发现两种选择的gRNA/Cas9 RNPs （gRNA5/Cas9, gRNA9/Cas9，特别是它们的组合）引起了最有效的抗病毒功效，这可以通过显著抑制HBV DNA， RNA和蛋白质的产生来证明。处理细胞的DNA测序显示，HBV靶基因的插入和删除（indel）或敲除（KO）突变率中等至高。基因比对分析显示，gRNA5和gRNA9靶序列在主要HBV基因型中均具有高度保守性，这表明这些基于crispr的基因编辑疗法具有靶向全球不同HBV毒株的潜力。因此，这些合成的gRNA/Cas9 RNPs代表了有希望的新疗法，可以开发和用于HBV基因破坏和病毒根除。

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引用次数: 0

Targeted inhibition of IGF2BP1 effectively suppresses HBV replication via an m6A-dependent manner 靶向抑制IGF2BP1通过m6a依赖的方式有效抑制HBV复制。

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2026-01-01 Epub Date: 2025-11-20 DOI: 10.1016/j.antiviral.2025.106310

Deyao Li , Yuxin Song , Danjuan Lu , Yanhua Zhang , Chenxiao Qu , Guiwen Guan , Yuexi Ma , Tianhao Mao , Liwei Zheng , Yi Wang , Wenhui Song , Ting Zhang , Fengmin Lu , Jing Ning , Xiangmei Chen

Elucidation of the regulation mechanism of hepatitis B virus (HBV) replication will provide potential targets for the development of novel anti-HBV therapeutics. It has been reported that the N6-methyladenosine (m6A) modification of HBV RNA plays a crucial role in the HBV life cycle. However, the mechanisms underlying the regulation of this modification remain incompletely understood. In this study, combining loss- and gain-of-function genetic analyses, we defined the role of IGF2BP1, an m6A reader, in facilitating HBV replication. Mechanistic studies revealed that IGF2BP1 stabilizes HBV RNAs primarily by binding to m6A-modified A1907 sites through its KH3-4 domain, thereby enhancing viral replication. Furthermore, targeted inhibition of IGF2BP1 by Cucurbitacin B, a small molecule inhibitor of IGF2BP1, was shown to inhibit HBV replication in vitro and in vivo. Taken together, these findings identify IGF2BP1 as a critical host regulator of HBV RNA stability through an m6A-dependent manner and targeted inhibition of IGF2BP1 effectively attenuates viral replication, providing a promising strategy for anti-HBV drug development.

阐明乙型肝炎病毒（HBV）复制的调控机制将为开发新型抗HBV治疗药物提供潜在靶点。据报道，HBV RNA的n6 -甲基腺苷（m6A）修饰在HBV生命周期中起着至关重要的作用。然而，这种修饰的调控机制仍然不完全清楚。在这项研究中，结合功能丧失和功能获得的遗传分析，我们确定了IGF2BP1（一种m6A读取器）在促进HBV复制中的作用。机制研究表明，IGF2BP1主要通过其KH3-4结构域结合m6a修饰的A1907位点来稳定HBV rna，从而增强病毒复制。此外，IGF2BP1的小分子抑制剂葫芦素B靶向抑制IGF2BP1，在体外和体内均可抑制HBV复制。综上所述，这些发现确定了IGF2BP1是HBV RNA稳定性的关键宿主调节剂，通过m6a依赖的方式和靶向抑制IGF2BP1有效地减弱病毒复制，为抗HBV药物开发提供了一个有希望的策略。在本研究中，我们发现宿主因子IGF2BP1通过其KH结构域，主要通过识别和结合HBV RNA上的m6A修饰位点来增强所有五种HBV RNA的稳定性，从而促进HBV RNA的表达和HBV复制。葫芦素B是IGF2BP1的小分子抑制剂，可通过降低HBV RNA表达有效抑制HBV复制，为抗HBV药物的开发提供了新的途径。

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引用次数: 0

Bulevirtide for chronic hepatitis delta: from clinical trials to real life data: an expert opinion report 布来韦肽治疗慢性丁型肝炎：从临床试验到现实生活数据：专家意见报告。

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2026-01-01 Epub Date: 2025-11-19 DOI: 10.1016/j.antiviral.2025.106311

Tarik Asselah , Homie Razavi , Hélène Fontaine , Kosh Agarwal

Hepatitis D virus (HDV) is a small RNA virus that requires Hepatitis B Surface Antigen (HBsAg) for its envelope. Eight genotypes with more than 80 % sequence homology and many subgenotypes have been described. Worldwide prevalence of chronic hepatitis delta (CHD) is estimated at about 5 % of chronic hepatitis B cases, translating to 15–20 million individuals. The diagnosis of HDV infection involves presence of antibodies to hepatitis D antigen (anti-HDV antibodies). Anti-HDV total antibody indicates HDV exposure (not infection). To document infection; the patient needs to undergo PCR testing and only if PCR is positive should the diagnosis of HDV ongoing infection done. Testing for the antibodies should be performed in all HBsAg-positive persons. CHD is more severe and progressive than HBV mono-infection, with a higher risk of cirrhosis and hepatocellular carcinoma (HCC), transplantation and death. Pegylated interferon-alpha (pegIFN-a) has been used for treating CHD with only limited durable responses. A 48-week course of weekly subcutaneous injections of pegIFN-a suppresses HDV replication in approximately 20–30 % of patients 24 weeks off therapy, with significant side effects. Bulevirtide (BLV) was approved by the European Medicines Agency (EMA) in 2020 for CHD and compensated liver disease. Since its approval, real-life data on the use of BLV have been accumulating, with most treated patients in Europe having advanced fibrosis or cirrhosis. Real life data efficacy is concordant to that seen in clinical trials, with many patients achieving significant reductions in HDV RNA levels and ALT normalization after several months of treatment, and favorable safety. However, HBsAg loss is relatively rare. Finite therapy of BLV, in combination with pegIFN-a, leads to significant durable response, with more than 30 % of patients achieving HDV RNA undetectability off therapy. We need new finite therapies. Further real-world data and newer therapies are required for this severe disease.

丁型肝炎病毒（HDV）是一种小RNA病毒，其包膜需要乙型肝炎表面抗原（HBsAg）。目前已发现8种序列同源性超过80%的基因型和许多亚基因型。据估计，全球慢性丁型肝炎（CHD）患病率约占慢性乙型肝炎病例的5%，相当于1500万至2000万人。丁型肝炎病毒感染的诊断包括存在丁型肝炎抗原抗体（抗丁型肝炎病毒抗体）。抗HDV总抗体表明HDV暴露（不是感染）。记录感染；患者需要进行聚合酶链反应检测，只有当聚合酶链反应呈阳性时，才应诊断为HDV持续感染。应对所有hbsag阳性的人进行抗体检测。冠心病比单HBV感染更严重和进展，肝硬化和肝细胞癌（HCC）、移植和死亡的风险更高。聚乙二醇化干扰素- α (pegIFN-a)已被用于治疗冠心病，但只有有限的持久反应。每周一次皮下注射pegIFN-a的48周疗程可以抑制约20-30%的患者在治疗24周后的HDV复制，但有明显的副作用。Bulevirtide （BLV）于2020年获得欧洲药品管理局（EMA）批准，用于治疗冠心病和代偿性肝病。自批准以来，BLV的实际使用数据一直在积累，欧洲大多数接受治疗的患者患有晚期纤维化或肝硬化。现实生活数据疗效与临床试验一致，许多患者在治疗几个月后HDV RNA水平和ALT正常化显著降低，安全性良好。然而，HBsAg损失相对罕见。有限的BLV治疗，结合pegIFN-a，导致显著的持久反应，超过30%的患者在治疗后达到HDV RNA不可检测。我们需要新的有限疗法。这种严重疾病需要进一步的真实世界数据和更新的治疗方法。

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引用次数: 0

Novel computational pipeline to identify target sites for broad spectrum antiviral drugs 新的计算管道确定广谱抗病毒药物的靶点。

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2026-01-01 Epub Date: 2025-12-01 DOI: 10.1016/j.antiviral.2025.106322

John D. Sears , Konstantin I. Popov , Paul A. Sylvester , Rebekah Dickmander , Jennifer Diaz , Che-Kang Chang , Julia Huff , Wes Sanders , Nicholas A. Saba , Madeleine Sorensen , Adam M. Drobish , Nicholas A. May , Kevin Namitz , Julia Fecko , Neela H. Yennawar , Thomas E. Morrison , Alexander Tropsha , Mark T. Heise , Nathaniel J. Moorman

Emerging viruses pose an ongoing threat to human health. While certain viral families are common sources of outbreaks, predicting the specific virus within a family that will cause the next outbreak or pandemic is not possible, creating an urgent need for broad spectrum antiviral drugs that are effective against a wide array of related viral pathogens. However, broad spectrum drug development is hindered by the lack of detailed knowledge of compound binding sites that are structurally and functionally conserved between viral family members and are essential for virus replication. To overcome this limitation, we developed an in silico approach that combines AI-driven protein structure prediction, computational fragment soaking, multiple sequence alignment, and protein stability calculations to identify highly conserved target sites that are both solvent-accessible and conserved. We applied this approach to the Togaviridae family, which includes emerging pandemic disease threats such as chikungunya and Venezuelan equine encephalitis virus for which there are currently no approved antiviral therapies. Our analysis identified multiple solvent accessible and structurally conserved pockets in the alphavirus non-structural protein 2 (nsP2) protease domain, which is essential for processing the viral replicase proteins. Mutagenesis of key solvent accessible and conserved residues identified novel pockets that are essential for the replication of multiple alphaviruses, validating these pockets as potential antiviral target sites for nsP2 inhibitors. These findings highlight the potential of artificial intelligence-informed modeling for revealing functionally conserved, accessible pockets as a means of identifying potential target binding sites for broadly active direct acting antivirals.

新出现的病毒对人类健康构成持续威胁。虽然某些病毒科是爆发的常见来源，但预测一个病毒科中的特定病毒将导致下一次爆发或大流行是不可能的，因此迫切需要对各种相关病毒病原体有效的广谱抗病毒药物。然而，由于缺乏对病毒家族成员之间结构和功能保守且对病毒复制至关重要的化合物结合位点的详细了解，广谱药物的开发受到阻碍。为了克服这一限制，我们开发了一种计算机方法，将人工智能驱动的蛋白质结构预测、计算片段浸泡、多序列比对和蛋白质稳定性计算相结合，以确定既可溶剂访问又保守的高度保守的靶点。我们将这种方法应用于托加病毒科，其中包括基孔肯雅病毒和委内瑞拉马脑炎病毒等新出现的大流行疾病威胁，目前尚无批准的抗病毒疗法。我们的分析在甲病毒非结构蛋白2 （nsP2）蛋白酶结构域中发现了多个溶剂可达和结构保守的口袋，这是处理病毒复制酶蛋白所必需的。对关键溶剂可及性和保守残基的诱变发现了对多种甲病毒复制至关重要的新口袋，验证了这些口袋作为nsP2抑制剂的潜在抗病毒靶点。这些发现突出了人工智能建模的潜力，揭示了功能保守的、可访问的口袋，作为识别广泛活性直接作用抗病毒药物的潜在目标结合位点的一种手段。

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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 : 2025-12-01 Epub 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 : 2025-12-01 Epub 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

The natural polyphenol proanthocyanidin A2 prevents the in vitro infection of Ebola virus and rabies virus by interfering with the early phases of the replication cycle 天然多酚原花青素A2通过干扰复制周期的早期阶段阻止埃博拉病毒和狂犬病毒的体外感染

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-12-01 Epub Date: 2025-11-20 DOI: 10.1016/j.antiviral.2025.106312

Denis Pasqual , Ilaria Artusi , Michele Paccagnella , Giulia Sibille , Mattia Mirandola , Sofia Appelberg , Maria Francesca Priore , Maira Zorzan , Massimo E. Maffei , Paola De Benedictis , Claudia Del Vecchio , Alì Mirazimi , Giorgio Cozza , Giorgio Gribaudo , Cristiano Salata

The Ebola virus (EBOV) and the Rabies virus (RABV) are deadly infectious agents impacting human and animal health. Current prevention and control strategies mainly rely on vaccines and antibodies, highlighting the urgent need for effective, low-cost antivirals suitable for therapeutic options. Plant-derived bioactive compounds offer a promising natural source for such candidate antivirals. As a contribution to this antiviral approach, we have characterized the anti-EBOV and anti-RABV activity of a Cranberry extract (CE) endowed with a very high content of bioactive A2-type proanthocyanidin (PAC-A2). The CE inhibited the in vitro infection of both pseudoviruses expressing EBOV-GP or RABV-G glycoproteins and authentic EBOV and RABV. Attachment and entry assays revealed that the extract targets early phases of infection preventing attachment and entry. Noteworthy, synthetic PAC-A2 reproduced the antiviral activity observed with the whole CE. Mechanistic studies then revealed that the CE interacted directly with the ectodomain of EBOV-GP or the RABV-G, suggesting interference with their functions. In support to this hypothesis, fluorescence spectroscopy analysis showed a reduction in intrinsic fluorescence of both EBOV-GP and RABV-G after incubation with synthetic PAC-A2, thus confirming a direct interaction of the viral glycoproteins with PAC-A2. In silico docking simulations further sustained in vitro results by predicting the binding of PAC-A2 into the binding pocket of EBOV-GP and to the trimeric architecture of RABV-G.

Together, these results suggest this cranberry extract and bioactive PAC-A2 as potential candidates to be further develop as novel antiviral agents for the prevention of EBOV and RABV infections.

埃博拉病毒（EBOV）和狂犬病毒（RABV）是影响人类和动物健康的致命传染病。目前的预防和控制战略主要依靠疫苗和抗体，这突出表明迫切需要适用于治疗方案的有效、低成本抗病毒药物。植物源性生物活性化合物为这类候选抗病毒药物提供了一个很有前途的天然来源。作为对这种抗病毒方法的贡献，我们已经鉴定了具有非常高含量的生物活性a2型原花青素（PAC-A2）的蔓越莓提取物（CE）的抗ebov和抗rabv活性。CE对表达EBOV- gp或RABV- g糖蛋白的假病毒和真EBOV和RABV的体外感染均有抑制作用。附着和进入试验表明，提取物针对感染的早期阶段，防止附着和进入。值得注意的是，合成PAC-A2复制了整个CE所观察到的抗病毒活性。机制研究表明，CE直接与EBOV-GP或RABV-G的外畴相互作用，表明其功能受到干扰。为了支持这一假设，荧光光谱分析显示，EBOV-GP和RABV-G与合成的PAC-A2一起培养后，其固有荧光减少，从而证实了病毒糖蛋白与PAC-A2的直接相互作用。通过预测PAC-A2与EBOV-GP结合口袋和RABV-G三聚体结构的结合，硅对接模拟进一步支持了体外结果。综上所述，这些结果表明蔓越莓提取物和具有生物活性的PAC-A2具有进一步开发的潜力，可作为预防EBOV和RABV感染的新型抗病毒药物。

{"title":"The natural polyphenol proanthocyanidin A2 prevents the in vitro infection of Ebola virus and rabies virus by interfering with the early phases of the replication cycle","authors":"Denis Pasqual , Ilaria Artusi , Michele Paccagnella , Giulia Sibille , Mattia Mirandola , Sofia Appelberg , Maria Francesca Priore , Maira Zorzan , Massimo E. Maffei , Paola De Benedictis , Claudia Del Vecchio , Alì Mirazimi , Giorgio Cozza , Giorgio Gribaudo , Cristiano Salata","doi":"10.1016/j.antiviral.2025.106312","DOIUrl":"10.1016/j.antiviral.2025.106312","url":null,"abstract":"<div><div>The Ebola virus (EBOV) and the Rabies virus (RABV) are deadly infectious agents impacting human and animal health. Current prevention and control strategies mainly rely on vaccines and antibodies, highlighting the urgent need for effective, low-cost antivirals suitable for therapeutic options. Plant-derived bioactive compounds offer a promising natural source for such candidate antivirals. As a contribution to this antiviral approach, we have characterized the anti-EBOV and anti-RABV activity of a Cranberry extract (CE) endowed with a very high content of bioactive A2-type proanthocyanidin (PAC-A2). The CE inhibited the <em>in vitro</em> infection of both pseudoviruses expressing EBOV-GP or RABV-G glycoproteins and authentic EBOV and RABV. Attachment and entry assays revealed that the extract targets early phases of infection preventing attachment and entry. Noteworthy, synthetic PAC-A2 reproduced the antiviral activity observed with the whole CE. Mechanistic studies then revealed that the CE interacted directly with the ectodomain of EBOV-GP or the RABV-G, suggesting interference with their functions. In support to this hypothesis, fluorescence spectroscopy analysis showed a reduction in intrinsic fluorescence of both EBOV-GP and RABV-G after incubation with synthetic PAC-A2, thus confirming a direct interaction of the viral glycoproteins with PAC-A2. <em>In silico</em> docking simulations further sustained <em>in vitro</em> results by predicting the binding of PAC-A2 into the binding pocket of EBOV-GP and to the trimeric architecture of RABV-G.</div><div>Together, these results suggest this cranberry extract and bioactive PAC-A2 as potential candidates to be further develop as novel antiviral agents for the prevention of EBOV and RABV infections.</div></div>","PeriodicalId":8259,"journal":{"name":"Antiviral research","volume":"244 ","pages":"Article 106312"},"PeriodicalIF":4.0,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145576257","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Genetic determinants of efficacy of antiviral drugs revealed by genome-wide CRISPR screens 全基因组CRISPR筛选揭示抗病毒药物疗效的遗传决定因素。

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-12-01 Epub Date: 2025-11-17 DOI: 10.1016/j.antiviral.2025.106309

Wei Jiang , Ailing Yang , Jingchuan Ma , Dawei Lv , Mingxian Liu , Chao Wang , Shuo Chen , Huiling Fang , Yankai Chu , Zhengjin He , Wenrui Li , Yucheng Liu , Yun Zhao , Zhaocai Zhou , Gang Long , Hai Jiang

Nucleoside and nucleobase analog antiviral drugs are pivotal in antiviral therapy, but comprehensive methods to understand their cellular response mechanisms and genetic regulators are still lacking. Here, we show that Eμ-Myc; Arf^−/− mouse lymphoma cells, which are highly apoptosis-prone, enabled genome-wide CRISPR-Cas9 screening on such drugs to identify genes that modulate their efficacy. Using retroviral sgRNA libraries and MAGeCK analysis, we uncovered key regulators of drug transport, activation, and inactivation for these drugs. For ribavirin, adenosine kinase (ADK) and adenylsuccinate synthase (ADSS) were critical for nucleotide metabolism and bioactivation. Remdesivir uptake and activation depended on the transporter SLC29A3 and phosphoamidase HINT1, whereas favipiravir resistance was linked to NT5C2-mediated dephosphorylation. Viral replication assays in Huh7 cells validated that knockout of SLC29A3, HINT1, or NT5C2 significantly altered antiviral efficacy. This study delineates the genetic network governing nucleotide analog response, providing mechanistic insights and potential biomarkers for personalized antiviral therapy.

核苷和核碱基类似物抗病毒药物是抗病毒治疗的关键，但了解其细胞反应机制和遗传调控因子的综合方法仍然缺乏。这里，我们证明了Eμ-Myc；Arf-/-小鼠淋巴瘤细胞是高度凋亡易感性的细胞，因此可以对这些药物进行全基因组CRISPR-Cas9筛选，以确定调节其疗效的基因。利用逆转录病毒sgRNA文库和MAGeCK分析，我们发现了这些药物转运、激活和失活的关键调控因子。对于利巴韦林，腺苷激酶（ADK）和腺苷琥珀酸合成酶（ADSS）是核苷酸代谢和生物活化的关键。瑞德西韦的摄取和激活依赖于转运体SLC29A3和磷酸化酶HINT1，而法匹拉韦耐药与nt5c2介导的去磷酸化有关。Huh7细胞中的病毒复制实验证实，敲除SLC29A3、HINT1或NT5C2显著改变了抗病毒效果。这项研究描述了控制核苷酸类似物反应的遗传网络，为个性化抗病毒治疗提供了机制见解和潜在的生物标志物。

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引用次数: 0