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A highly potent and stable pan-coronavirus fusion inhibitor as a candidate prophylactic and therapeutic for COVID-19 and other coronavirus diseases

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Acta Pharmaceutica Sinica B 2022年第4期12卷 1652-1661页

作者： Jie Zhou Wei Xu zezhong liu Chao Wang Shuai Xia Qiaoshuai Lan Yanxing Cai Shan Su Jing Pu Lixiao Xing Youhua Xie Lu Lu Shibo Jiang Qian Wang Key Laboratory of Medical Molecular Virology(MOE/NHC/CAMS) School of Basic Medical Sciences and BSL-3 FacilityShanghai Institute of Infectious Diseases and BiosecurityFudan UniversityShanghai 200032China State Key Laboratory of Toxicology and Medical Countermeasures Beijing Institute of Pharmacology and ToxicologyBeijing 100850China

The development of broad-spectrum antivirals against human coronaviruses(HCoVs)is critical to combat the current coronavirus disease 2019(COVID-19)pandemic caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)and its variants,as well as future outbreaks of emerging CoVs.We have previously identified a polyethylene glycol-conjugated(PEGylated)lipopeptide,EK1C4,with potent pan-CoV fusion inhibitory activity.However,PEG linkers in peptide or protein drugs may reduce stability or induce anti-PEG antibodies in vivo.Therefore,we herein report the design and synthesis of a series of dePEGylated lipopeptide-based pan-CoV fusion inhibitors featuring the replacement of the PEG linker with amino acids in the heptad repeat 2 C-terminal fragment(HR2-CF)of HCoV-OC43.Among these lipopeptides,EKL1C showed the most potent inhibitory activity against infection by SARS-CoV-2 and its spike(S)mutants,as well as other HCoVs and some bat SARS-related coronaviruses(SARSr-CoVs)tested.The dePEGylated lipopeptide EKL1C exhibited significantly stronger resistance to proteolytic enzymes,better metabolic stability in mouse serum,higher thermostability than the PEGylated lipopeptide EK1C4,suggesting that EKL1C could be further developed as a candidate prophylactic and therapeutic for COVID-19 and other coronavirus diseases.

关键词： Coronavirus Lipopeptide SARS-CoV-2 Polyethylene glycol Fusion inhibitor

Inefficiency of Sera from Mice Treated with Pseudotyped SARS-CoV to Neutralize 2019-nCoV Infection

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Virologica Sinica 2020年第3期35卷 340-343页

作者： zezhong liu Shuai Xia Xinling Wang Qiaoshuai Lan Wei Xu Qian Wang Shibo Jiang Lu Lu Key Laboratory of Medical Molecular Virology(MOE/NHC/CAMS) School of Basic Medical SciencesFudan UniversityShanghai 200032China

Dear Editor,An outbreak of unusual pneumonia in Wuhan, China recently was caused by infection of a novel type of coronavirus. The virus and disease were denoted as 2019-nCoV and COVID-19, respectively, by the World Health Organization(WHO). Most recently, 2019-nCoV was renamed SARS-CoV-2 by Coronaviridae Study Group(CSG) of the International Committee on Taxonomy of Viruses(ICTV)(Gorbalenya et al. 2020)。

关键词： al. infection Wuhan

基于重组人Ⅲ型胶原蛋白的三聚体抗原疫苗策略在新冠和流感疫苗中的应用

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合成生物学 2024年第2期5卷 385-395页

作者：刘泽众周洁朱赟陆路姜世勃复旦大学基础医学院上海200032 复旦大学药学院药理系上海201203 中国科学院生物物理研究所北京100101

新冠病毒等多种包膜病毒的糖蛋白在天然状态下呈现三聚体形式。三聚体蛋白通常较单体蛋白具有更优的免疫原性。目前使用非人蛋白来源的Foldon等三聚体基序可促使目的蛋白形成稳定三聚体,但这些基序的强免疫原性会削弱目的蛋白的免疫应答... 详细信息

新冠病毒等多种包膜病毒的糖蛋白在天然状态下呈现三聚体形式。三聚体蛋白通常较单体蛋白具有更优的免疫原性。目前使用非人蛋白来源的Foldon等三聚体基序可促使目的蛋白形成稳定三聚体,但这些基序的强免疫原性会削弱目的蛋白的免疫应答,限制了其在疫苗抗原设计中的应用。在本研究中,发现并利用重组人源化Ⅲ型胶原蛋白(Rh3C)作为新型三聚体基序,利用合成生物学方法,与新冠病毒刺头(spike)蛋白受体结合域(RBD)融合表达后促使RBD形成三聚体。这种胶原-RBD(Rh3C-RBD)三聚体蛋白在免疫小鼠后,较RBD单体蛋白诱导产生了更高滴度的结合抗体和中和抗体。此外,我们还进一步验证了该Rh3C基序与流感病毒HA1蛋白融合后,较HA1单体蛋白可在小鼠体内诱导更强效的抗体应答。因此,这种新型的Rh3C基序,有望广泛用于三聚体疫苗抗原的设计和优化。

关键词：三聚体抗原人胶原蛋白新冠病毒流感病毒疫苗设计

Repurposing of a clinically used anti-HPV agent to prevent and treat SARS-CoV-2 infection as an intranasal formulation

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Signal Transduction and Targeted Therapy 2021年第9期6卷 2631-2633页

作者： Chen Hua Qinhai Ma Yun Zhu Shuai Xia zezhong liu Lin Li Lu Lu Nanshan Zhong Shuwen liu Zifeng Yang Shibo Jiang Key Laboratory of Medical Molecular Virology(MOE/NHC/CAMS) School of Basic Medical SciencesShanghai Institute of Infectious Disease and BiosecurityFudan UniversityShanghaiChina State Key Laboratory of Respiratory Disease National Clinical Research Center for Respiratory DiseaseGuangzhou Institute of Respiratory Healththe First Affiliated Hospital of Guangzhou Medical UniversityGuangzhouChina National Laboratory of Biomacromolecules Institute of BiophysicsChinese Academy of SciencesBeijingChina State Key Laboratory of Organ Failure Research Guangdong Provincial Key Laboratory of New Drug ScreeningSchool of Pharmaceutical SciencesSouthern Medical UniversityGuangzhouChina Guangzhou Laboratory Bio-IslandGuangzhouChina

Dear Editor,Fan et al.have shown that whey protein,such as that found in human,bovine,and goat milk,could effectively inhibit SARS-CoV-2 infection,but they did not identify the components in whey protein having this activity.1 Since p-lactoglobulin(P-LG)is the main component of whey protein,we herein tested the anti-SARS-CoV-2 activity of p-LG and 3-hydroxyphthalic anhydride(3HP)-modified P-LG(3HP-P-LG),an active component in anti-human papillomavirus(HPV)biological dressing(JB01-BD)that has been used in clinics since 2013 to block cervical infection of HPV.

关键词： infection nasal

An ultrapotent pan-β-coronavirus lineage B(β-CoV-B)neutralizing antibody locks the receptor-binding domain in closed conformation by targeting its conserved epitope

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Protein & Cell 2022年第9期13卷 655-675页

作者： zezhong liu Wei Xu Zhenguo Chen Wangjun Fu Wuqiang Zhan Yidan Gao Jie Zhou Yunjiao Zhou Jianbo Wu Qian Wang Xiang Zhang Aihua Hao Wei Wu Qianqian Zhang Yaming Li Kaiyue Fan Ruihong Chen Qiaochu Jiang Christian TMayer Till Schoofs Youhua Xie Shibo Jiang Yumei Wen Zhenghong Yuan Kang Wang Lu Lu Lei Sun Qiao Wang Key Laboratory of Medical Molecular Virology(MOE/NHC/CAMS) School of Basic Medical SciencesShanghai Institute of Infectious Disease and Biosecuritythe Fifth People’s Hospital of ShanghaiShanghai Key Laboratory of Medical EpigeneticsInternational Co-laboratory of Medical Epigenetics and Metabolism(Ministry of Science and Technology)Institutes of Biomedical SciencesBiosafety Level 3 LaboratoryShanghai Medical CollegeFudan UniversityShanghai 200032China CAS Key Laboratory of Infection and Immunity National Laboratory of MacromoleculesInstitute of BiophysicsChinese Academy of SciencesBeijing 100101China Experimental Immunology Branch Center for Cancer ResearchNational Cancer InstituteNational Institutes of HealthBethesdaMD 20892USA GSK Vaccines 1300 WavreBelgium

New threats posed by the emerging circulating variants of SARS-CoV-2 highlight the need to find conserved neutralizing epitopes for therapeutic antibodies and efficient vaccine design.Here,we identified a receptorbinding domain(RBD)-binding antibody,XG014,which potently neutralizesβ-coronavirus lineage B(β-CoV-B),including SARS-CoV-2,its circulating variants,SARSCoV and bat SARSr-CoV WIV1.Interestingly,antibody family members competing with XG014 binding show reduced levels of cross-reactivity and induce antibodydependent SARS-CoV-2 spike(S)protein-mediated cellcell fusion,suggesting a unique mode of recognition by XG014.Structural analyses reveal that XG014 recognizes a conserved epitope outside the ACE2 binding site and completely locks RBD in the non-functional“down”conformation,while its family member XG005 directly competes with ACE2 binding and position the RBD“up”.Single administration of XG014 is effective in protection against and therapy of SARS-CoV-2 infection in vivo.Our findings suggest the potential to develop XG014 as pan-β-CoV-B therapeutics and the importance of the XG014 conserved antigenic epitope for designing broadly protective vaccines againstβ-CoV-B and newly emerging SARS-CoV-2 variants of concern.

关键词： SARS-CoV-2 neutralizing antibody receptor-binding domain XG014 antibody-dependent cellcell fusion

The role of furin cleavage site in SARS-CoV-2 spike protein-mediated membrane fusion in the presence or absence of trypsin

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Signal Transduction and Targeted Therapy 2020年第1期5卷 1630-1632页

作者： Shuai Xia Qiaoshuai Lan Shan Su Xinling Wang Wei Xu zezhong liu Yun Zhu Qian Wang Lu Lu Shibo Jiang Key Laboratory of Medical Molecular Virology(MOE/NHC/CAMS) School of Basic Medical SciencesFudan University130 Dong An RoadShanghai 200032China National Laboratory of Biomacromolecules Institute of BiophysicsChinese Academy of SciencesBeijing 100101China

Dear Editor,The rapid spread of SARS-CoV-2(also known as 2019-nCoV and HCoV-191),a novel lineage B betacoronavirus(βCoV),has caused a global pandemic of coronavirus disease(COVID-19).It has been speculated that RRAR,a unique furin-like cleavage site(FCS)in the spike protein(S),which is absent in other lineage BβCoVs,such as SARS-CoV,is responsible for its high infectivity and transmissibility.

关键词： cleavage absence trypsin

RBD-Fc-based COVID-19 vaccine candidate induces highly potent SARS-CoV-2 neutralizing antibody response

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Signal Transduction and Targeted Therapy 2020年第1期5卷 196-205页

作者： zezhong liu Wei Xu Shuai Xia Chenjian Gu Xinling Wang Qian Wang Jie Zhou Yanling Wu Xia Cai Di Qu Tianlei Ying Youhua Xie Lu Lu Zhenghong Yuan Shibo Jiang Key Laboratory of Medical Molecular Virology(MOE/NHC/CAMS) School of Basic Medical Sciences and BSL-3 facilityFudan UniversityShanghai 200032China

The pandemic of coronavirus disease 2019(COVID-19)caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)has posed serious threats to global health and economy,thus calling for the development of safe and effective vaccines.The receptorbinding domain(RBD)in the spike protein of SARS-CoV-2 is responsible for its binding to angiotensin-converting enzyme 2(ACE2)receptor.It contains multiple dominant neutralizing epitopes and serves as an important antigen for the development of COVID-19 vaccines.Here,we showed that immunization of mice with a candidate subunit vaccine consisting of SARS-CoV-2 RBD and Fc fragment of human IgG,as an immunopotentiator,elicited high titer of RBD-specific antibodies with robust neutralizing activity against both pseudotyped and live SARS-CoV-2 infections.The mouse antisera could also effectively neutralize infection by pseudotyped SARS-CoV-2 with several natural mutations in RBD and the IgG extracted from the mouse antisera could also show neutralization against pseudotyped SARS-CoV and SARS-related coronavirus(SARSr-CoV).Vaccination of human ACE2 transgenic mice with RBD-Fc could effectively protect mice from the SARS-CoV-2 challenge.These results suggest that SARS-CoV-2 RBD-Fc has good potential to be further developed as an effective and broad-spectrum vaccine to prevent infection of the current SARS-CoV-2 and its mutants,as well as future emerging SARSr-CoVs and re-emerging SARS-CoV.

关键词： ACE2 vaccine neutral

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不同手术方式治疗老年胸腰椎结核疗效对比

潍坊医学院学报 2024年第2期46卷 111-113页

作者：王国良刘湘泽刘泽众孙立磊潍坊市人民医院关节外科山东潍坊261041 潍坊市第二人民医院骨科不详

目的比较前后联合入路一期后路固定、二期前路病灶切除、钛笼植骨融合与一期病灶切除、钛笼植骨融合并椎弓根螺钉治疗老年胸腰椎结核的临床效果及安全性。方法选取2017年1月~2020年12月在潍坊市第二人民医院收治的老年胸腰结核患者30... 详细信息

目的比较前后联合入路一期后路固定、二期前路病灶切除、钛笼植骨融合与一期病灶切除、钛笼植骨融合并椎弓根螺钉治疗老年胸腰椎结核的临床效果及安全性。方法选取2017年1月~2020年12月在潍坊市第二人民医院收治的老年胸腰结核患者30例。A组行一期病灶切除、钛笼植骨融合并椎弓根螺钉内固定(15例);B组行一期后路固定、二期前路病灶切除、钛笼自体骨植骨融合手术(15例)。抗结核治疗贯穿整个治疗过程。分别对两组患者的手术情况、住院恢复情况、并发症发生情况以及生活质量进行比较。结果 (1)B组患者的手术所使用的时间、术中的出血量以及术后引流量均显著少于A组,差异有统计学意义(P0.05);(4)术后两组患者的QOL评分、KPS评分比较,差异均无统计学意义(P>0.05);两组患者随访期间均未见结核复发以及椎弓根螺钉断裂等严重并发症,CT复查均显示骨性融合。结论不同手术方式治疗老年胸腰椎结核均取得较好的临床效果,前后联合入路一期后路固定、二期前路病灶切除、钛笼植骨融合治疗老年胸腰椎结核,在减少手术时间、术中出血量具有显著优势,在老年患者中可进行选择。

关键词：手术方式老年胸腰椎结核治疗效果

RETRACTED ARTICLE:SARS-CoV-2 infects T lymphocytes through its spike protein-mediated membrane fusion

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Cellular & Molecular Immunology 2023年第5期20卷 554-554页

作者： Xinling Wang Wei Xu Gaowei Hu Shuai Xia Zhiping Sun zezhong liu Youhua Xie Rong Zhang Shibo Jiang Lu Lu Key Laboratory of Medical Molecular Virology(MOE/NHC/CAMS) School of Basic Medical Sciences and Biosafety Level 3 LaboratoryFudan UniversityShanghaiChina Lindsley F.Kimball Research Institute New York

The authors have retracted this article [1]. After the publication ofthis article, it came to the authors attention that in order tosupport the conclusions of the study, the authors should haveused primary T cells instead of T cell lines. Additionally, there areconcerns that the flow cytometry methodology applied here wasflawed. These points resulted in the conclusions being consideredinvalid.

关键词： conclusions lymphocytes ART