Antiviral research最新文献_第7页

Efficient eukaryotic expression and potent antiviral activity of a long-acting recombinant feline interferon-ω2-Fc fusion protein against major feline viruses 一种长效重组猫干扰素ω2- fc融合蛋白的高效真核表达及抗病毒活性研究。

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-09-03 DOI: 10.1016/j.antiviral.2025.106272

Yuwei Yang , Hetao Song , Ke Zhang , Siyin Wang , Ya Zhao , Qiang Zhang , Meilin Jin

Feline interferon-ω2 (FeIFN-ω2) holds potential as a therapeutic agent against feline viral infections. However, its clinical application is limited by rapid clearance and suboptimal antiviral effectiveness. Thus, in this study, an Fc-fused construct, FeIFN-ω2-Fc, was engineered to improve antiviral potency and pharmacokinetic properties both in vitro and in vivo. Three recombinant constructs—native FeIFN-ω2, FeIFN-ω2-Dimer, and FeIFN-ω2-Fc—were expressed in Chinese Hamster Ovary cells. All showed strong antiviral activity (10^6.74–10^8.42 IU/mg) and effectively activated downstream interferon signaling. Functional assays, including 50 % tissue culture infectious dose assay, quantitative polymerase chain reaction, and immunofluorescence, confirmed their ability to inhibit feline calicivirus (FCV), feline herpesvirus type 1 (FHV-1), and feline parvovirus (FPV). Glycosylation analysis revealed two sites (S102 and T128) in the ω2 domain of FeIFN-ω2-Fc that contributed to its structural stability and functional enhancement. Among the three candidates, FeIFN-ω2-Fc demonstrated the best overall profile, with higher expression levels, simplified purification, and favorable pharmacokinetics. In animal models, it was well tolerated and significantly alleviated clinical symptoms, reduced viral loads, and preserved tissue integrity following FHV-1 infection. Pharmacokinetic studies showed a marked increase in plasma half-life, from 5.80 ± 1.75 h for the native protein to 34.05 ± 6.36 h for the Fc-fused form. Further extension to 40.55 ± 6.61 h was achieved by introducing YTE mutations (S250Y/S252T/T254E) within the Fc region. Based on these findings, a dosing regimen of 4 × 10⁵ IU/kg every other day is proposed, supporting FeIFN-ω2-Fc as a strong candidate for feline antiviral therapy.

猫干扰素-ω - 2 （FeIFN-ω - 2）具有治疗猫病毒感染的潜力。然而，其临床应用受到快速清除和非最佳抗病毒效果的限制。因此，在本研究中，我们设计了一种fc融合构建物FeIFN-ω2-Fc，以提高体外和体内的抗病毒效力和药代动力学特性。在中国仓鼠卵巢细胞中表达了FeIFN-ω - 2、FeIFN-ω - 2-二聚体和FeIFN-ω - 2- fc三种重组结构。所有药物均表现出较强的抗病毒活性（106.74 ~ 108.42 IU/mg），并有效激活下游干扰素信号。功能试验，包括50%组织培养感染剂量试验、定量聚合酶链反应和免疫荧光，证实了它们抑制猫杯状病毒（FCV）、猫疱疹病毒1型（FHV-1）和猫细小病毒（FPV）的能力。糖基化分析发现FeIFN-ω2-Fc的ω2域有两个位点（S102和T128），这两个位点有助于其结构稳定性和功能增强。其中，FeIFN-ω - 2- fc具有较高的表达水平、纯化过程简化、良好的药代动力学等特点。在动物模型中，它耐受性良好，显著缓解了FHV-1感染后的临床症状，降低了病毒载量，并保持了组织完整性。药代动力学研究显示血浆半衰期明显增加，从天然蛋白的5.80±1.75小时增加到fc融合形式的34.05±6.36小时。通过在Fc区引入YTE突变（S250Y/S252T/T254E），进一步延长至40.55±6.61 h。基于这些发现，建议每隔一天4 × 105 IU/kg的给药方案，支持FeIFN-ω - 2- fc作为猫抗病毒治疗的强有力候选者。

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引用次数: 0

Structural determinants of nervous system exposure of adibelivir (IM-250) and related herpes helicase-primase inhibitors across animal species 神经系统暴露的结构决定因素adibelivir （IM-250）和相关的疱疹解旋酶启动酶抑制剂在动物物种。

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-09-03 DOI: 10.1016/j.antiviral.2025.106271

Christian Gege, Thomas Hoffmann, Gerald Kleymann

The high incidence and prevalence of herpes infections pose a significant health burden worldwide. Herpes simplex virus infections are the cause of herpes labialis, genital herpes or herpes keratitis and in rare cases life-threatening herpes encephalitis, meningitis or disseminated disease. After primary infection, herpes simplex viruses (HSVes) establish latency in the trigeminal and sacral ganglia and at least 30 % of patients experience clinically manifestant recurrences for life. For effective treatment of these neurotrophic HSVes, adequate drug exposure in the nervous system is essential.

Here we report the post administration exposure of structurally different helicase-primase inhibitors (HPIs) in plasma, blood, organs and, in particular, the nervous system of animals by HPLC/MS. In diverse animal species, after single or multiple doses of helicase-primase drugs by oral or intravenous administration, only adibelivir (IM-250) achieved concentrations in the nervous system in the range of plasma or blood levels (ratio 0.5 to 4 nervous system/plasma), while other helicase-primase inhibitors with distinct structures, including amenamevir, pritelivir or ABI-5366, showed a low brain/plasma ratio of less than 0.1. The efficient passage of helicase-primase drugs through the blood-brain and blood-nerve barrier is based on their distinct structure and chemical properties. In preclinical studies published so far, adibelivir was efficacious in the herpes encephalitis and neonatal animal model and reduced the reactivation competence of the neuronal latent herpes viral reservoir. Ongoing clinical trials with HPIs will show whether sufficient drug exposure in brain and ganglia will translate into more effective herpes therapies for patients.

疱疹感染的高发病率和流行率在世界范围内构成了重大的健康负担。单纯疱疹病毒感染会引起唇疱疹、生殖器疱疹或角膜炎，在极少数情况下还会引起危及生命的疱疹性脑炎、脑膜炎或播散性疾病。原发性感染后，单纯疱疹病毒（HSVes）在三叉神经节和骶神经节建立潜伏期，至少30%的患者经历临床明显的复发。为了有效治疗这些神经营养性HSVes，在神经系统中充分的药物暴露是必不可少的。在这里，我们报告了结构不同的解旋酶引物酶抑制剂（hpi）在给药后暴露于血浆、血液、器官，特别是神经系统中的HPLC/MS。在多种动物中，单次或多次口服或静脉给药解旋酶引物酶药物后，只有adibelivir （IM-250）在神经系统中的浓度达到血浆或血液水平范围内（神经系统/血浆的比率为0.5至4），而其他结构不同的解旋酶引物酶抑制剂，包括阿莫那韦、priitelivir或ABI-5366，其脑/血浆的比率低于0.1。解旋酶引物酶药物有效通过血脑和血神经屏障是基于它们独特的结构和化学性质。在目前发表的临床前研究中，阿迪贝利韦在疱疹脑炎和新生儿动物模型中有效，并降低了神经元潜伏疱疹病毒库的再激活能力。正在进行的hpi临床试验将显示在大脑和神经节中充分的药物暴露是否会转化为对患者更有效的疱疹治疗。

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引用次数: 0

DMBT1 promotes SARS-CoV-2 infection and its SRCR-derived peptide inhibits SARS-CoV-2 infection DMBT1促进SARS-CoV-2感染，其srcr衍生肽抑制SARS-CoV-2感染。

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-09-03 DOI: 10.1016/j.antiviral.2025.106269

Chenxi Zhu , Ziqiao Wang , Zhendong Pan , Xinjia Mai , Yiyun Chen , Wen Zhang , Ping Zhao , Hailin Tang , Rong Zhang , Dapeng Zhou

DMBT1 is a large scavenger receptor cysteine rich (SRCR) B protein that has been reported as a tumor suppressor gene and a co-receptor for HIV-1 infection. Here, we found DMBT1 is a major mucosal protein bound to SARS-CoV-2. Overexpression of DMBT1 in 293T cells may enhanced infection by SARS-CoV-2 in ACE2 dependent manner. Blocking experiments using overlapping peptide library of SRCR domain of DMBT1 showed that peptide 7 (CQGRVEVLYRGSWGTV), which contains bacteria-binding VEVLXXXXW motif, could inhibit SARS-CoV-2 infection. High concentration of peptide 7 can significantly inhibit the replication of SARS-CoV-2 in hamsters. Peptide 7 inhibits SARS-CoV-2 infection by aggregating the spike protein, thereby reducing its binding to and internalization by host cells. The cysteine residue at the N-terminus of peptide 7 is critical for dimerization and antiviral activity. These results indicate that DMBT1 can serve as a candidate target for the development of antiviral drugs.

DMBT1是一种富含半胱氨酸的大清道夫受体（SRCR） B蛋白，已被报道为肿瘤抑制基因和HIV-1感染的共受体。在这里，我们发现DMBT1是与SARS-CoV-2结合的主要粘膜蛋白。293T细胞中DMBT1的过表达可能以ACE2依赖的方式增强SARS-CoV-2感染。利用DMBT1 SRCR结构域重叠肽文库的阻断实验表明，含有细菌结合的VEVLXXXXW基序的肽7 （CQGRVEVLYRGSWGTV）可抑制SARS-CoV-2感染。高浓度肽7能显著抑制SARS-CoV-2在仓鼠体内的复制。肽7通过聚集刺突蛋白抑制SARS-CoV-2感染，从而减少其与宿主细胞的结合和内化。肽7 n端的半胱氨酸残基对二聚化和抗病毒活性至关重要。这些结果表明，DMBT1可以作为开发抗病毒药物的候选靶点。

引用次数: 0

Ensitrelvir suppresses prolonged olfactory abnormalities derived from SARS-CoV-2 infection in hamsters 恩司替韦抑制仓鼠SARS-CoV-2感染引起的长时间嗅觉异常。

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-09-02 DOI: 10.1016/j.antiviral.2025.106270

Ryoichi Tashima, Takayuki Kuroda, Haruaki Nobori, Satoshi Miyagawa, Takuya Yamane, Alice Shimba, Masaaki Nakashima, Keita Fukao

Ensitrelvir, an oral severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) 3CL protease inhibitor, is reportedly effective in suppressing smell disorder onset, a post-coronavirus disease 2019 (COVID-19) condition symptom. However, the pathogenesis of post-COVID-19 condition symptoms and the mechanism underlying the onset-suppressive effect of ensitrelvir are not fully understood. Here, we explored a post-COVID-19 condition model in hamsters 1 month post-SARS-CoV-2 infection and showed that ensitrelvir treatment caused early recovery of body weight, viral RNA suppression, and sense of smell improvement. In the nasal turbinates, SARS-CoV-2 was associated with significantly increased inflammatory markers, many of which were suppressed by ensitrelvir. Significant positive correlations were observed between smell testing and many inflammation-related markers in the nasal turbinates. In conclusion, our study indicates that chronic inflammation may occur in the nasal turbinates over a long period post-SARS-CoV-2 infection, leading to smell disorder onset in a hamster model. Early ensitrelvir treatment post-infection suppressed inflammation in the nasal turbinates and prevented smell disorder onset.

据报道，Ensitrelvir是一种口服严重急性呼吸综合征冠状病毒2 (SARS-CoV-2) 3CL蛋白酶抑制剂，可有效抑制2019年冠状病毒病（COVID-19）后症状嗅觉障碍的发作。然而，covid -19后症状的发病机制和ensitrelvir抑制发病作用的机制尚不完全清楚。在这里，我们探索了感染sars - cov -2后1个月的仓鼠后covid -19状态模型，结果表明，恩司替韦治疗可导致体重早期恢复、病毒RNA抑制和嗅觉改善。在鼻鼻甲中，SARS-CoV-2与炎症标志物显著增加相关，其中许多炎症标志物被恩司替韦抑制。嗅觉测试与鼻甲骨中许多炎症相关标志物之间观察到显著的正相关。总之，我们的研究表明，在sars - cov -2感染后的很长一段时间内，鼻甲可能会发生慢性炎症，导致仓鼠模型出现嗅觉障碍。感染后早期恩司替韦治疗可抑制鼻甲炎症，防止嗅觉障碍发作。

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引用次数: 0

Ubiquitinated hepatitis D antigen-induced CD8+ T-cell responses inhibit HDV replication in HDV-infected liver organoids 泛素化丁型肝炎抗原诱导的CD8+ t细胞反应抑制丙型肝炎病毒感染的肝类器官中的丙型肝炎病毒复制

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-09-02 DOI: 10.1016/j.antiviral.2025.106266

Leer Shen , Luying Tian , Qingxin Guo , Rongli He , Yi Zhang , Siyuan Ma , Weiwei Hu , Jie Chen , Soon Seng Ng , Jinmei Chen , Xiaohua Chen

Background

Hepatitis D virus (HDV) infection is the most severe form of human viral hepatitis. A poor virus-specific CD8⁺T cell response may result in persistent HDV infection. We investigated anti-viral effect and mechanisms of ubiquitinated small hepatitis D antigen (Ub-S-HDAg) in HBV/HDV superinfected liver organoids.

Methods

Induced pluripotent stem cells were differentiated into hepatocyte-like cells and seeded onto inverted colloidal crystal scaffolds. A Ub-S-HDAg recombinant lentiviral plasmid was constructed and transduced into dendritic cells (DCs), which were then co-cultured with T cells. Cytokine levels secreted by CD8⁺T cells and their antiviral efficacy were evaluated, and transcriptomic sequencing was conducted to elucidate immunoregulatory mechanisms of Ub-S-HDAg.

Results

We successfully established a model that stably expressed HDV. Ub-S-HDAg significantly promoted DC maturation and reduced HDV RNA viral load and HDAg levels. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis and gene set enrichment analyses suggested that DCs transduced with Ub-S-HDAg promoted CD8⁺ T cell activation via Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway. JAK1, JAK2, STAT1, and STAT4 expressions in Ub-S-HDAg group increased and was blocked by JAK2 inhibitor AG490. Additionally, Ub-S-HDAg induced DC maturation-enhanced cytotoxic T lymphocyte responses and increased IFN-γ, IL-2, IL-12, and TNF-α secretion.

Conclusion

We successfully developed HBV/HDV superinfected liver organoids and suggest that Ub-S-HDAg facilitates DC maturation, inducing CD8⁺ T cell proliferation through the JAK/STAT signalling pathway, leading to a stronger antiviral immune response.

丁型肝炎病毒（HDV）感染是人类病毒性肝炎最严重的形式。病毒特异性CD8+T细胞反应差可能导致持续的HDV感染。我们研究了泛素化丁型肝炎小抗原（Ub-S-HDAg）在HBV/HDV重复感染的肝类器官中的抗病毒作用及其机制。方法将诱导多能干细胞分化为肝细胞样细胞，植入倒置胶体晶体支架。构建Ub-S-HDAg重组慢病毒质粒，转染树突状细胞（dc），与T细胞共培养。评估CD8+T细胞分泌的细胞因子水平及其抗病毒效果，并通过转录组测序阐明Ub-S-HDAg的免疫调节机制。结果成功建立了稳定表达HDV的模型。Ub-S-HDAg显著促进DC成熟，降低HDV RNA病毒载量和HDAg水平。京都基因和基因组百科全书通路富集分析和基因集富集分析表明，用Ub-S-HDAg转导的dc通过Janus kinase (JAK)/signal transducator and activator of transcription （STAT）通路促进CD8+ T细胞活化。JAK1、JAK2、STAT1和STAT4在Ub-S-HDAg组的表达升高，并被JAK2抑制剂AG490阻断。此外，Ub-S-HDAg诱导DC成熟增强细胞毒性T淋巴细胞反应，增加IFN-γ、IL-2、IL-12和TNF-α分泌。我们成功开发了HBV/HDV超感染的肝类器官，并提示Ub-S-HDAg促进DC成熟，通过JAK/STAT信号通路诱导CD8+ T细胞增殖，导致更强的抗病毒免疫应答。

{"title":"Ubiquitinated hepatitis D antigen-induced CD8+ T-cell responses inhibit HDV replication in HDV-infected liver organoids","authors":"Leer Shen , Luying Tian , Qingxin Guo , Rongli He , Yi Zhang , Siyuan Ma , Weiwei Hu , Jie Chen , Soon Seng Ng , Jinmei Chen , Xiaohua Chen","doi":"10.1016/j.antiviral.2025.106266","DOIUrl":"10.1016/j.antiviral.2025.106266","url":null,"abstract":"<div><h3>Background</h3><div>Hepatitis D virus (HDV) infection is the most severe form of human viral hepatitis. A poor virus-specific CD8<sup>+</sup>T cell response may result in persistent HDV infection. We investigated anti-viral effect and mechanisms of ubiquitinated small hepatitis D antigen (Ub-S-HDAg) in HBV/HDV superinfected liver organoids.</div></div><div><h3>Methods</h3><div>Induced pluripotent stem cells were differentiated into hepatocyte-like cells and seeded onto inverted colloidal crystal scaffolds. A Ub-S-HDAg recombinant lentiviral plasmid was constructed and transduced into dendritic cells (DCs), which were then co-cultured with T cells. Cytokine levels secreted by CD8<sup>+</sup>T cells and their antiviral efficacy were evaluated, and transcriptomic sequencing was conducted to elucidate immunoregulatory mechanisms of Ub-S-HDAg.</div></div><div><h3>Results</h3><div>We successfully established a model that stably expressed HDV. Ub-S-HDAg significantly promoted DC maturation and reduced HDV RNA viral load and HDAg levels. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis and gene set enrichment analyses suggested that DCs transduced with Ub-S-HDAg promoted CD8<sup>+</sup> T cell activation via Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway. JAK1, JAK2, STAT1, and STAT4 expressions in Ub-S-HDAg group increased and was blocked by JAK2 inhibitor AG490. Additionally, Ub-S-HDAg induced DC maturation-enhanced cytotoxic T lymphocyte responses and increased IFN-γ, IL-2, IL-12, and TNF-α secretion.</div></div><div><h3>Conclusion</h3><div>We successfully developed HBV/HDV superinfected liver organoids and suggest that Ub-S-HDAg facilitates DC maturation, inducing CD8<sup>+</sup> T cell proliferation through the JAK/STAT signalling pathway, leading to a stronger antiviral immune response.</div></div>","PeriodicalId":8259,"journal":{"name":"Antiviral research","volume":"243 ","pages":"Article 106266"},"PeriodicalIF":4.0,"publicationDate":"2025-09-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144989912","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

Comprehensive insights into Japanese encephalitis virus: From molecular characterization to advanced detection and vaccine strategies 全面了解日本脑炎病毒：从分子表征到先进的检测和疫苗策略。

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-09-02 DOI: 10.1016/j.antiviral.2025.106268

Pooja Chugh , Subodh Soni , Nisha Ghanghas , Sachin Kumar , Hari Mohan

The Japanese encephalitis virus (JEV) remains a major cause of viral encephalitis in Asia, with significant morbidity and mortality. This review offers a comprehensive overview of the current landscape of JEV research, focusing on its genomic structure, protein composition, and global epidemiology. We highlight the complexity of JEV transmission and pathogenesis, examining the interplay of demographic factors and geographic spread. In particular, we assess the evolution of diagnostic methodologies from traditional molecular and serological techniques to emerging biosensor-based approaches, emphasizing advancements in sensitivity and rapidity. The application of CRISPR/Cas systems for JEV detection marks a promising frontier in molecular diagnostics. Additionally, we review the current status of JEV vaccines, discussing recent innovations in vaccine development aimed at enhancing immunogenicity and accessibility. Beyond prevention, a spectrum of antiviral strategies—including direct-acting antivirals, entry inhibitors, host-directed modulators, neuroprotective agents, and steroidal/synthetic compounds—has demonstrated potent in vitro and in vivo efficacy, targeting viral enzymes, structural proteins, and host pathways. This review underscores the critical role of advanced detection strategies and vaccines in controlling JEV, offering insights into ongoing efforts to mitigate its impact in endemic regions.

日本脑炎病毒（JEV）仍然是亚洲病毒性脑炎的主要病因，发病率和死亡率很高。本文综述了目前乙脑病毒的研究概况，重点介绍了乙脑病毒的基因组结构、蛋白质组成和全球流行病学。我们强调乙脑病毒传播和发病机制的复杂性，研究人口因素和地理传播的相互作用。特别是，我们评估了诊断方法的演变，从传统的分子和血清学技术到新兴的基于生物传感器的方法，强调了灵敏度和快速的进步。CRISPR/Cas系统在乙脑病毒检测中的应用是分子诊断领域的一个有前景的前沿。此外，我们回顾了乙脑疫苗的现状，讨论了旨在提高免疫原性和可及性的疫苗开发的最新创新。除了预防之外，一系列抗病毒策略——包括直接作用的抗病毒药物、进入抑制剂、宿主定向调节剂、神经保护剂和甾体/合成化合物——已经证明了有效的体外和体内疗效，靶向病毒酶、结构蛋白和宿主途径。这一综述强调了先进的检测策略和疫苗在控制乙脑病毒方面的关键作用，为减轻其在流行地区影响的持续努力提供了见解。

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引用次数: 0

A novel synthetic bile acid derivative inhibits hepatitis B virus infection at entry step by interfering with the oligomerization of sodium taurocholate co-transporting polypeptide 一种新的合成胆汁酸衍生物通过干扰牛磺胆酸钠共转运多肽的寡聚化抑制乙型肝炎病毒的进入步骤感染

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-08-30 DOI: 10.1016/j.antiviral.2025.106267

Gede Ngurah Rsi Suwardana , Takayuki Abe , Lin Deng , Chieko Matsui , Takashi Okitsu , Takeshi Yamada , Manabu Hatano , Pattama Wiriyasermkul , Shushi Nagamori , Sameh A. Gad , Hussein H. Aly , Hironori Nishitsuji , Kunitada Shimotohno , Ikuo Shoji

Hepatitis B virus (HBV) infection is a major global health burden worldwide despite the availability of an effective vaccine and effective anti-HBV drugs. The currently approved anti-HBV drugs—i.e., nucleos(t)ide analogues and pegylated interferon α—can effectively suppress HBV replication, but rarely achieve a functional cure. Accordingly, new anti-HBV agents targeting different aspects of the HBV life cycle are needed. In this study, we screened for anti-HBV agents using the recombinant HBV expressing NanoLuc (NL) reporter gene (HBV/NL) and our original synthetic heterocyclic compound library. As a result, we identified a synthetic bile acid derivative, SO-145, as a potential novel anti-HBV agent, and investigated its effects in several cellular models of HBV. Treatment of HepG2-NTCP-C4 cells with SO-145 suppressed their NL activity following infection with HBV/NL. SO-145 suppressed HBV replication in PXB-cells infected with HBV genotype D, but did not show any inhibitory effect on HBV replication in Hep38.7-Tet cells. These results suggest that SO-145 specifically inhibits the early phase of the HBV life cycle. In other experiments, SO-145 was also shown to inhibit hepatitis D virus infection. Immunofluorescence analysis using fluorescent-labeled preS1 peptide revealed that SO-145 does not inhibit the preS1 attachment to the NTCP, but does markedly inhibit the HBV/preS1 internalization. Moreover, SO-145 does not inhibit the bile acid uptake facilitated by NTCP. Further mechanistic analysis suggested that SO-145 interferes with the NTCP oligomerization. Taken together, these results suggest that SO-145 inhibits HBV entry into hepatocytes by interfering with the NTCP oligomerization.

尽管有有效的疫苗和有效的抗HBV药物，但乙型肝炎病毒（HBV）感染仍是全球主要的健康负担。目前已批准的抗hbv药物，即例如，核苷(t)类似物和聚乙二醇化干扰素α -可以有效抑制HBV复制，但很少实现功能性治愈。因此，需要针对HBV生命周期不同方面的新型抗HBV药物。在这项研究中，我们使用表达NanoLuc （NL）报告基因的重组HBV （HBV/NL）和我们的原始合成杂环化合物文库筛选抗HBV药物。因此，我们确定了一种合成胆汁酸衍生物SO-145，作为一种潜在的新型抗HBV药物，并研究了其在几种HBV细胞模型中的作用。用SO-145治疗HepG2-NTCP-C4细胞可抑制HBV/NL感染后的NL活性。SO-145对感染HBV基因型D的pxb细胞的HBV复制有抑制作用，但对Hep38.7-Tet细胞的HBV复制无抑制作用。这些结果表明，SO-145特异性抑制HBV生命周期的早期阶段。在其他实验中，SO-145也显示出抑制丁型肝炎病毒感染的作用。使用荧光标记preS1肽的免疫荧光分析显示，SO-145不抑制preS1附着在NTCP上，但明显抑制HBV/preS1内化。此外，SO-145不抑制NTCP促进的胆汁酸摄取。进一步的机制分析表明，SO-145干扰NTCP寡聚化。综上所述，这些结果表明SO-145通过干扰NTCP寡聚化抑制HBV进入肝细胞。

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引用次数: 0

Characterization of remdesivir resistance mutations in COVID-19 patients with various immunosuppressive diseases 不同免疫抑郁性疾病的COVID-19患者瑞德西韦耐药突变特征

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-08-28 DOI: 10.1016/j.antiviral.2025.106264

Takaya Ichikawa , Tomokazu Tamura , Naganori Nao , Hikoyu Suzuki , Shuhei Maruyama , Daiki Wada , Shuhei Tsujino , Noriyoshi Yoshinaga , Kohsuke Asagoe , Mutsumi Takahata , Takashi Ishio , Makoto Ibata , Tanino Yoko , Yasutaka Kakinoki , Kazuhiro Okubo , Rigel Suzuki , Saori Suzuki , Yasushi Nakamori , Takanori Teshima , Takasuke Fukuhara

Immunocompromised patients (ICPs), such as those who receive certain immunosuppressive therapies, occasionally experience prolonged viral infections even after antiviral treatment. In some cases, antiviral-resistant viruses may eventually emerge. Remdesivir (RDV) is an adenosine nucleoside analog that inhibits the activity of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA polymerase, which is composed of a catalytic subunit known as nsp12. Previous studies have reported that amino acid mutations around loci 790–810 in nsp12 are responsible for RDV resistance. However, the host immune status that promotes the emergence of resistant viruses and their virological features remain unclear. We therefore collected clinical samples from 15 coronavirus disease 2019 (COVID-19) patients with various immunosuppressive conditions who received RDV. Variant analysis identified a total of seven nsp12 mutations—V792I, M794I, E796D, E796K, C799F, C799Y, and T803I—in 80 % (12/15) of the participants, with M794I and V792I the most prevalent. The identified mutations were more frequently observed in severely ICPs (including patients with hematological malignancies or kidney transplantation) compared to non-severely ICPs (including patients with solid cancers or autoimmune diseases). In vitro analysis using recombinant viruses carrying each identified mutation demonstrated that all mutant viruses were less efficient in growth compared to wildtype but with a 1.8-fold (T803I) to 3.6-fold (V792I) increase in the half maximal effective concentration (EC₅₀) of RDV. The present study highlights the importance of monitoring resistance mutations to anti–SARS-CoV-2 drugs in severely ICPs.

免疫功能低下患者（icp），如接受某些免疫抑制治疗的患者，即使在抗病毒治疗后，偶尔也会经历长期的病毒感染。在某些情况下，抗病毒病毒最终可能出现。Remdesivir （RDV）是一种腺苷核苷类似物，可抑制严重急性呼吸综合征冠状病毒2 (SARS-CoV-2) RNA聚合酶的活性，该酶由催化亚基nsp12组成。先前的研究报道了nsp12中790-810位点附近的氨基酸突变与RDV抗性有关。然而，促进耐药病毒出现的宿主免疫状态及其病毒学特征尚不清楚。因此，我们收集了15例接受RDV治疗的不同免疫抑制条件的2019冠状病毒病（COVID-19）患者的临床样本。变异分析在80%（12/15）的参与者中发现了七种nsp12突变——V792I、M794I、E796D、E796K、C799F、C799Y和t803i，其中M794I和V792I最为普遍。与非严重icp患者（包括实体癌或自身免疫性疾病患者）相比，在严重icp患者（包括血液系统恶性肿瘤或肾移植患者）中发现的突变更频繁。体外分析表明，携带每种突变的重组病毒与野生型相比，所有突变病毒的生长效率都较低，但RDV的一半最大有效浓度（EC50）增加了1.8倍（T803I）至3.6倍（V792I）。本研究强调了监测严重icp患者对抗sars - cov -2药物耐药突变的重要性。

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引用次数: 0

An engineered chimeric ACE2-HR2 peptide exhibits potent and broad-spectrum activity against SARS-CoV-2 variants 一种工程嵌合ACE2-HR2肽对SARS-CoV-2变体显示出有效的广谱活性

IF 4 2区医学 Q1 PHARMACOLOGY & PHARMACY

Antiviral research

Pub Date : 2025-08-28 DOI: 10.1016/j.antiviral.2025.106265

Wenwen Bi , Tao Zhu , Yawen Xu , Yihui Chen , Jianmin Li

The persistent circulation of SARS-CoV-2 and limited efficacy of current vaccines in preventing viral transmission underscore the critical need for broad-spectrum antivirals. Here, we report a novel engineering strategy to develop A3M6L35HR2 (FL), a cholesterol-free recombinant peptide derived from our previously reported A1L35HR2, by incorporating an optimized angiotensin-converting enzyme 2 (ACE2)-mimetic peptide A3M6 with six residue mutations. A3M6L35HR2 (FL) exhibited superior inhibitory potency against diverse SARS-CoV-2 variants with IC₅₀ from 1.7 to 16.2 nM, including antigenically distinct Omicron sublineages, achieving 2- to 29-fold enhanced activity compared to A1L35HR2. A3M6L35HR2 (FL) exhibits a higher α-helicity (70 %) and stronger binding affinity (K_D = 0.03 nM) with the viral HR1 domain, thereby effectively suppressing spike-mediated membrane fusion at nanomolar concentrations. Combining A3M6L35HR2 (FL) with RBD-targeting protein T-ACE2 exhibited antagonistic effect, attributed to competitive RBD binding by both agents. These findings establish A3M6L35HR2 (FL) as a promising antiviral candidate against current and emerging SARS-CoV-2 variants.

SARS-CoV-2的持续传播和现有疫苗在预防病毒传播方面的有限效力，强调了对广谱抗病毒药物的迫切需要。在这里，我们报告了一种新的工程策略来开发A3M6L35HR2 (FL)，这是一种无胆固醇重组肽，来源于我们之前报道的A1L35HR2，通过结合优化的血管紧张素转换酶2 (ACE2)-模拟肽A3M6具有六个残基突变。A3M6L35HR2 （FL）对多种SARS-CoV-2变体表现出优异的抑制能力，IC50为1.7至16.2 nM，包括抗原性不同的Omicron亚系，与A1L35HR2相比，活性增强了2至29倍。A3M6L35HR2 （FL）具有较高的α-螺旋度（70%）和与病毒HR1结构域较强的结合亲和力（KD = 0.03 nM），从而在纳摩尔浓度下有效抑制峰介导的膜融合。将A3M6L35HR2 （FL）与RBD靶向蛋白T-ACE2联合使用显示出拮抗作用，这归因于两种药物与RBD的竞争性结合。这些发现确定A3M6L35HR2 （FL）是一种有希望的抗病毒候选药物，可用于对抗当前和新出现的SARS-CoV-2变体。

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引用次数: 0

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