VP35

摘要非分段负链RNA病毒（nsNSV），包括埃博拉病毒（EBOV）、狂犬病病毒、人呼吸道合胞病毒和肺病毒，可引起人类和动物的呼吸道感染、出血热和脑炎，并被视为全球重大的健康和经济负担。病毒基因组的复制和转录由大L聚合酶执行，L聚合酶是开发抗病毒药物的有希望的靶标。2023年9月12日，中国科学院微生物研究所施一、高福及齐建勋共同通讯在Nature 在线发表题为“Molecular mechanism of de novo replication by the Ebola virus polymerase”的研究论文，该研究揭示了埃博拉病毒聚合酶从头复制的分子机制。该研究以EBOV L聚合酶为代表，在酶促实验中发现，L聚合酶的novo复制受EBOV基因组特异性3′先导序列控制，至少3个碱基可以有效地驱动RNA合成的延伸过程，而不依赖于特异性RNA序列。该研究确定了EBOV L-VP35-RNA复合物的高分辨率结构，并发现3 '先导RNA以独特的稳定弯曲构象结合在模板进入通道中。进一步的突变研究证实了RNA的弯曲构象是RNA从头复制活性所必需的，并揭示了L蛋白中稳定RNA构象的关键残基。这些发现为nsNSV聚合酶的RNA合成提供了新的机制，并揭示了抗病毒药物开发的重要靶点。负链RNA病毒(NSVs)包括许多高度流行的人类和动物病原体，如埃博拉病毒(EBOV)、拉沙病毒(LASV)、尼帕病毒、狂犬病病毒(RABV)、流感病毒和人类呼吸道合胞病毒(HRSV)。NSVs被认为是对全球经济和公共卫生的重大威胁。因此，更好地了解NSV复制的分子机制和开发新的抗NSV感染的抗病毒药物对于对抗这些病毒至关重要。根据病毒基因组的组织结构，NSVs可进一步分为包括EBOV在内的非节段NSVs (nsNSVs)和包括流感病毒在内的节段NSVs (sNSVs)。nsNSVs和sNSVs的单链RNA基因组都与病毒核蛋白组装成螺旋构型的病毒核糖核蛋白(vRNP)复合物。在sNSVs中，每个基因组病毒RNA (vRNA) 片段可以通过在同一病毒群中高度保守的部分互补的5 '和3 '末端形成一个“伪环”。而对于nsNSVs, vRNA的5 ' 和3 '端通常分别位于相反的两端。所有NSVs编码含有RNA依赖性RNA 聚合酶(RdRP)结构域的大RNA聚合酶，并在vRNPs的背景下复制或转录vRNA基因组。在过去的几年中，作者对sNSVs的复制和转录机制获得了实质性的结构见解，包括正黏液病毒A病毒、B病毒、C病毒和D病毒。许多sNSV RNA聚合酶的结构表明，RNA聚合酶通常具有一个保守的RdRP核心，表现为具有拇指、手指和手掌亚结构域的右手状褶皱，以及其他几个功能结构域，包括核酸内切酶结构域和帽结合结构域。EBOV L-VP35-RNA复合物的整体结构（图源自Nature ）nsNSVs的聚合酶在基因组3′端启动转录，先导区和内部基因依次转录成先导RNA和具有5′-cap和3′-poly (A)尾结构的单顺反子mRNA。据推测，nsNSVs的转录遵循一种启停机制，通过识别每个病毒基因侧面的基因起始(GS) 和基因结束(GE)信号。每个转录本在各自的GE信号处终止后，聚合酶要么与模板分离，要么在其GS信号处继续下游转录本的合成。对于基因组复制，聚合酶也在基因组的3 '端启动，产生全长补体基因组RNA，忽略所有GS和GE信号。关于聚合酶如何从转录转换到复制的分子机制尚不清楚，这种转换可能受到P蛋白磷酸化或其他转录激活因子状态的调节。目前，在nsNSV RNA聚合酶复合物的可用结构中未观察到RNA结合。这些聚合酶如何识别基因组和抗基因组RNA中的末端启动子元件仍然是难以捉摸的。先前的研究表明，VSV和RABV L蛋白的N端残基以及P蛋白的C端结构域可能参与病毒RNA的结合，因为它们位于模板RNA进入通道附近。然而，准确的聚合酶-RNA相互作用尚不清楚，这极大地阻碍了人们对nsNSVs RNA复制和转录事件的起始机制的分子理解。EBOV作为非nsNSV的代表，对人类具有高致病性，可引起出血热和多器官衰竭等严重临床症状。EBOV RNP复合体负责病毒RNA合成，而L-VP35复合体是有效RNA聚合活性的基本功能单元。最近，利用冷冻电镜(cryo-EM)方法解决了EBOV L-VP复合物的整体结构，并清楚地显示了L蛋白中与辅因子VP35四聚体紧密相互作用的活性位点的结构特征。然而，对于识别其vRNA启动子的L蛋白的功能和结构特征知之甚少，这阻碍了对EBOV复制从头起始过程的分子理解。总之，该研究提供了EBOV L聚合酶识别基因组RNA的第一个结构细节，并揭示了nsNSV复制起始的独特机制。这些发现将极大地增强人们对nsNSV RNA聚合酶合成RNA的机制的理解，并为开发针对nsNSV聚合酶的新型抗病毒药物开辟道路。原文链接：https://www-nature-com.libproxy1.nus.edu.sg/articles/s41586-023-06608-1今年是COVID-19大流行趋于稳定后的第一年，人民生活逐渐走向正轨，经济也在缓慢但有力的复苏之中，而各抗体企业也依然在不断奋进、不断突破、不断超越，使抗体领域蓬勃发展，硕果累累。为促进抗体行业的交流与创新，2023年10月14-15日第六届金秋十月抗体产业发展大会如约而至。会议旨在为研究人员提供一个互动交流的平台，有助于推动抗体产业的进一步发展。会议内容时间：2023年10月14-15日地点：苏州（酒店定向通知）规模：600-800人主办单位：生物制品圈、抗体圈指导单位：康维讯生物、夏尔巴生物会议费用：免费FREE!（仅收取50元报名定金，含参会学习、茶歇、会议手册，定金概不退还），9月15日报名定金提高到100元，先到先得，报完即止，超过9月20日预登记将收取会议费！报名方式：扫描下方二维码或点击文章最底部“阅读原文”→ 填写表格 → 报名成功（报名志愿者，承担一定工作，请慎重考虑，免交定金）！组委会获得报名信息后，根据报名信息进行初筛，并进一步与报名者沟通确认，实现精准邀请。最终有机会进入大会微信群（严格审核通过）。日程安排更多嘉宾仍在邀请中。。。。。。识别微信二维码，添加生物制品圈小编，符合条件者即可加入生物制品微信群！请注明：姓名+研究方向！版权声明本公众号所有转载文章系出于传递更多信息之目的，且明确注明来源和作者，不希望被转载的媒体或个人可与我们联系(cbplib@163.com)，我们将立即进行删除处理。所有文章仅代表作者观点，不代表本站立场。

Growing body of data supports Ampligen’s potential as both a prophylactic and an early-onset therapy against human Ebola virus disease outbreaksOCALA, Fla., March 16, 2023 (GLOBE NEWSWIRE) -- AIM ImmunoTech Inc. (NYSE American: AIM) (“AIM” or the “Company”), an immuno-pharma company focused on the research and development of therapeutics to treat multiple types of cancers, immune disorders, and viral diseases — including COVID-19, the disease caused by the SARS-CoV-2 virus — today announced a late-breaking presentation at the 36th International Conference on Antiviral Research (ICAR) detailing Ampligen’s mechanism of action in the treatment of Ebola virus disease (EVD). The presentation was given by Angela Corona, PhD, Department of Life and Environmental Sciences, University of Cagliari, Monserrato, Italy, and one of the published authors of a recent manuscript titled “Ebola virus disease: In vivo protection provided by the PAMP restricted TLR3 agonist rintatolimod and its mechanism of action.” Ebola virus (EBOV) is a highly infectious and lethal pathogen responsible for sporadic, self-limiting clusters of EVD in Central Africa capable of reaching epidemic status. Highlights of the recently published Ampligen data that was presented include: As a TLR3 agonist, Ampligen induces and enhances innate immunological responses to EBOV infection.Ampligen appears to inactivate the EBOV lethal factor (EBOV VP35), which is believed to be responsible, in part, for the high mortality rate observed in humans, by acting as a “competitive decoy.” VP35 is understood to sequester the dsRNA produced by EBOV during its replication, which inhibits the normal innate immune responses to viral infection.Ampligen protected mice from a lethal challenge by mouse adapted EBOV-Zaire and its associated weight loss.The 6 mg/kg doses, frequently used in AIM’s clinical trials, used in mouse model is easily achievable and well tolerated in humans.Overall, the data suggest Ampligen’s potential as a viable candidate to protect against exposure to EBOV. The corresponding slides from the presentation at ICAR are now available on the Company’s website (aimimmuno.com) on the “Events and Presentations” page. AIM CEO Thomas K. Equels commented, “In vivo pre-clinical analysis shows that Ampligen has potential as both a prophylactic and an early-onset therapeutic in Ebola virus disease. Animal studies with Ampligen at the United States Army Medical Research Institute of Infectious Diseases Biosafety Level 4 laboratories demonstrated 100% protective survival, as compared to 100% mortality in Ebola virus disease. Those animal studies are indicative of the drug’s high potential for efficacy. The analysis presented at ICAR now provides a clear understanding of why Ampligen is achieving such a high level of efficacy in these animal models, signaling its potential as a human therapy.” Ampligen has been granted Orphan Drug status for Ebola virus disease by the European Medicines Agency and the FDA. Ampligen is not currently approved for the prevention or treatment of EVD. The Company intends to file Investigational New Drug applications to study the use of Ampligen (1) as an early onset therapy for the treatment of EBOV, and (2) for the prevention of Ebola virus reactivation. Ultimately, the Company's goal is to seek a Tropical Disease Priority Review Voucher for new drug applications associated with these indications. About the International Society for Antiviral Research (ISAR) The International Society for Antiviral Research (ISAR) is an internationally recognized organization for scientists involved in basic, applied, and clinical aspects of antiviral research. The Society main event is the annual International Conference on Antiviral Research (ICAR), a truly interdisciplinary meeting which attracts the interest of chemists, biologists, and clinicians. About Ampligen Ampligen is AIM’s dsRNA product candidate being developed for globally important cancers, viral diseases and disorders of the immune system. Ampligen is currently being evaluated as a combinational therapy for the treatment of a variety of solid tumor types in multiple clinical trials – both underway and planned – at major cancer research centers around the country. Ampligen is being used to treat pancreatic cancer patients in an Early Access Program approved by the Inspectorate of Healthcare in the Netherlands at Erasmus Medical Center. Additionally, Ampligen is also approved in Argentina for the treatment of severe chronic fatigue syndrome and is currently being evaluated in SARS-CoV-2/COVID-19, myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) and Post COVID Conditions. About AIM ImmunoTech Inc. AIM ImmunoTech Inc. is an immuno-pharma company focused on the research and development of therapeutics to treat multiple types of cancers, immune disorders, and viral diseases, including COVID-19. The Company’s lead product, an investigational new drug called Ampligen® (rintatolimod) is an immuno-modulator with broad spectrum activity being investigated for globally important cancers, viral diseases and disorders of the immune system. For more information, please visit aimimmuno.com and connect with the Company on Twitter, LinkedIn, and Facebook. Cautionary Statement This press release contains forward-looking statements within the meaning of the Private Securities Litigation Reform Act of 1995 (the “PSLRA”). Words such as “may,” “will,” “expect,” “plan,” “anticipate” and similar expressions (as well as other words or expressions referencing future events or circumstances) are intended to identify forward-looking statements. Many of these forward-looking statements involve a number of risks and uncertainties. The clinical study and other matters discussed above are subject to a number of unknowns including, but not limited to, that significant additional testing and trials will be required to determine whether Ampligen will be an effective treatment for Ebola virus, comments and additional testing requested by the FDA if and when the Company files the Investigational New Drug application and whether the Company will be able to obtain a Tropical Disease Priority Review Voucher for the new drug application. Among other things, for those statements, the Company claims the protection of safe harbor for forward-looking statements contained in the PSLRA. The Company does not undertake to update any of these forward-looking statements to reflect events or circumstances that occur after the date hereof. Two of the authors of the manuscript are officers or directors of the Company. The information found on our website is not incorporated by reference into this press release and is included for reference purposes only.

OCALA, Fla., March 08, 2023 (GLOBE NEWSWIRE) -- AIM ImmunoTech Inc. (NYSE American: AIM) (“AIM” or the “Company”), an immuno-pharma company focused on the research and development of therapeutics to treat multiple types of cancers, immune disorders, and viral diseases — including COVID-19, the disease caused by the SARS-CoV-2 virus — today announced the publication of a new analysis of the ability of AIM’s drug Ampligen (rintatolimod) to inhibit the spread and replication of Ebola virus disease, which adds to the body of evidence supporting Ampligen’s potential as an early-onset prophylactic therapy against human Ebola outbreaks. The data were published in the peer-reviewed journal Antiviral Research, in a manuscript titled, “Ebola virus disease: In vivo protection provided by the PAMP restricted TLR3 agonist rintatolimod and its mechanism of action” and is now available on the Company’s website on the “Events and Presentations” page. Ebola virus (EBOV) is a highly infectious and lethal pathogen responsible for sporadic, self-limiting clusters of Ebola virus disease (EVD) in Central Africa capable of reaching epidemic status. Previously announced data found that Ampligen provided 100% protection against EVD in a mouse model. The newly published analysis supports a dual mechanism of action when Ampligen is used as a prophylactic therapy against EVD. As a TLR3 agonist, the drug induces and enhances the body’s innate immunological responses to EBOV infection. Additionally, Ampligen appears to inactivate the EBOV lethal factor (EBOV VP35), which is believed to be responsible, in part, for the high mortality rate observed in humans, by acting as a “competitive decoy.” VP35 is understood to sequester the dsRNA produced by EBOV during its replication, which inhibits the normal innate immune responses to viral infection. These analyses provide supporting evidence to help explain Ampligen’s ability to provide protection against EVD in a mouse model at the generally well-tolerated doses frequently used in AIM’s clinical trials. Overall, the study data and new analyses give AIM additional confidence in Ampligen’s potential as a viable candidate to protect against exposure to EBOV. David Strayer, MD, AIM’s Chief Scientific and Medical Officer and one of the published manuscript’s authors, commented “100% prophylactic protection has been shown in an Ebola mouse infection model at Ampligen doses easily achievable in humans. The study also confirms that Ampligen’s activity, unlike other dsRNAs, excludes the activation of the inflammatory RIG-1 cytosolic helicases.” Ampligen is not currently approved for the prevention or treatment of EVD. Ampligen was granted Orphan Drug status for Ebola virus disease by the European Medicines Agency in 2015, which means that, upon commercial approval, AIM will receive benefits including ten years of protection from market competition with similar medicines in the European Union. Additionally, in November of last year, the company announced that the U.S. Food and Drug Administration had also granted Orphan Drug Designation to Ampligen for EVD, which among other incentives includes seven years of market exclusivity after approval. Additionally, AIM intends to New Drug applications to study the use of Ampligen (1) as an early onset therapy for the treatment of EBOV, and (2) for the prevention of Ebola virus reactivation. Ultimately, the Company's goal is to seek a Tropical Disease Priority Review Voucher for new drug applications associated with these indications. About Ampligen Ampligen is AIM’s dsRNA product candidate being developed for globally important cancers, viral diseases and disorders of the immune system. Ampligen is currently being evaluated as a combinational therapy for the treatment of a variety of solid tumor types in multiple clinical trials – both underway and planned – at major cancer research centers around the country. Ampligen is being used to treat pancreatic cancer patients in an Early Access Program approved by the Inspectorate of Healthcare in the Netherlands at Erasmus Medical Center. Additionally, Ampligen is also approved in Argentina for the treatment of severe chronic fatigue syndrome and is currently being evaluated in SARS-CoV-2/COVID-19, myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) and Post COVID Conditions. About AIM ImmunoTech Inc. AIM ImmunoTech Inc. is an immuno-pharma company focused on the research and development of therapeutics to treat multiple types of cancers, immune disorders, and viral diseases, including COVID-19. The Company’s lead product, an investigational new drug called Ampligen® (rintatolimod) is an immuno-modulator with broad spectrum activity being investigated for globally important cancers, viral diseases and disorders of the immune system. For more information, please visit aimimmuno.com and connect with the Company on Twitter, LinkedIn, and Facebook. Cautionary Statement This press release contains forward-looking statements within the meaning of the Private Securities Litigation Reform Act of 1995 (the “PSLRA”). Words such as “may,” “will,” “expect,” “plan,” “anticipate” and similar expressions (as well as other words or expressions referencing future events or circumstances) are intended to identify forward-looking statements. Many of these forward-looking statements involve a number of risks and uncertainties. The clinical study and other matters discussed above are subject to a number of unknowns including, but not limited to, that significant additional testing and trials will be required to determine whether Ampligen will be an effective treatment for Ebola virus, comments and additional testing requested by the FDA if and when the Company files the Investigational New Drug application and whether the Company will be able to obtain a Tropical Disease Priority Review Voucher for the new drug application. Among other things, for those statements, the Company claims the protection of safe harbor for forward-looking statements contained in the PSLRA. The Company does not undertake to update any of these forward-looking statements to reflect events or circumstances that occur after the date hereof. Two of the authors of the manuscript are officers or directors of the Company. The information found on our website is not incorporated by reference into this press release and is included for reference purposes only. Investor Relations Contact JTC Team, LLC Jenene Thomas 833-475-8247 AIM@jtcir.com