title: Preparation of monoPEGylated Cyanovirin-N's derivative and its anti-influenza A virus bioactivity in vitro and in vivo.
abstract: Influenza A virus (IAV) has been raising public health and safety concerns worldwide. Cyanovirin-N (CVN) is a prominent anti-IAV candidate, but both cytotoxicity and immunogenicity have hindered the development of this protein as a viable therapy. In this article, linker-CVN (LCVN) with a flexible and hydrophilic polypeptide at the N-terminus was efficiently produced from the cytoplasm of Escherichia coli at a >15-l scale. PEGylation at the N-terminal α-amine of LCVN was also reformed as 20 kDa PEGylated linkered Cyanovirin-N (PEG20k-LCVN). The 50% effective concentrations of PEG20k-LCVN were 0.43 ± 0.11 µM for influenza A/HK/8/68 (H3N2) and 0.04 ± 0.02 µM for A/Swan/Hokkaido/51/96 (H5N3), dramatically lower than that of the positive control, Ribavirin (2.88 ± 0.66 × 10(3) µM and 1.79 ± 0.62 × 10(3) µM, respectively). A total of 12.5 µM PEG20k-LCVN effectively inactivate the propagation of H3N2 in chicken embryos. About 2.0 mg/kg/day PEG20k-LCVN increased double the survival rate (66.67%, P = 0.0378) of H3N2 infected mice, prolonged the median survival period, downregulated the mRNA level of viral nuclear protein and decreased (attenuated) the pathology lesion in mice lung. A novel PEGylated CVN derivative, PEG20k-LCVN, exhibited potent and strain-dependent anti-IAV activity in nanomolar concentrations in vitro, as well as in micromolar concentration in vivo.
authors.affiliation: Institute of Biomedicine & National Engineering Research Center of Genetic Medicine, Department of Cellular Biology, Jinan University, Guangzhou 510632, Guangdong, People's Republic of China; Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Centre for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China; Department of Pharmacy, College of Food and Pharmacy & Medical, Zhejiang Ocean University, Zhoushan 316002, People's Republic of China; Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, People's Republic of China; Division of Molecular Pharmacology of Infectious Agents, Department of Molecular Microbiology and Immunology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki City, Nagasaki Prefecture 852-8521, Japan; and Department of Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA Institute of Biomedicine & National Engineering Research Center of Genetic Medicine, Department of Cellular Biology, Jinan University, Guangzhou 510632, Guangdong, People's Republic of China; Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Centre for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China; Department of Pharmacy, College of Food and Pharmacy & Medical, Zhejiang Ocean University, Zhoushan 316002, People's Republic of China; Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, People's Republic of China; Division of Molecular Pharmacology of Infectious Agents, Department of Molecular Microbiology and Immunology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki City, Nagasaki Prefecture 852-8521, Japan; and Department of Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA.
authors.forename: Chongchao
authors.initials: C
authors.lastname: Wu
authors.affiliation: Institute of Biomedicine & National Engineering Research Center of Genetic Medicine, Department of Cellular Biology, Jinan University, Guangzhou 510632, Guangdong, People's Republic of China; Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Centre for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China; Department of Pharmacy, College of Food and Pharmacy & Medical, Zhejiang Ocean University, Zhoushan 316002, People's Republic of China; Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, People's Republic of China; Division of Molecular Pharmacology of Infectious Agents, Department of Molecular Microbiology and Immunology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki City, Nagasaki Prefecture 852-8521, Japan; and Department of Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA.
authors.forename: Wei
authors.initials: W
authors.lastname: Chen
authors.affiliation: Institute of Biomedicine & National Engineering Research Center of Genetic Medicine, Department of Cellular Biology, Jinan University, Guangzhou 510632, Guangdong, People's Republic of China; Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Centre for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China; Department of Pharmacy, College of Food and Pharmacy & Medical, Zhejiang Ocean University, Zhoushan 316002, People's Republic of China; Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, People's Republic of China; Division of Molecular Pharmacology of Infectious Agents, Department of Molecular Microbiology and Immunology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki City, Nagasaki Prefecture 852-8521, Japan; and Department of Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA.
authors.forename: Jia
authors.initials: J
authors.lastname: Chen
authors.affiliation: Institute of Biomedicine & National Engineering Research Center of Genetic Medicine, Department of Cellular Biology, Jinan University, Guangzhou 510632, Guangdong, People's Republic of China; Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Centre for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China; Department of Pharmacy, College of Food and Pharmacy & Medical, Zhejiang Ocean University, Zhoushan 316002, People's Republic of China; Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, People's Republic of China; Division of Molecular Pharmacology of Infectious Agents, Department of Molecular Microbiology and Immunology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki City, Nagasaki Prefecture 852-8521, Japan; and Department of Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA Institute of Biomedicine & National Engineering Research Center of Genetic Medicine, Department of Cellular Biology, Jinan University, Guangzhou 510632, Guangdong, People's Republic of China; Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Centre for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China; Department of Pharmacy, College of Food and Pharmacy & Medical, Zhejiang Ocean University, Zhoushan 316002, People's Republic of China; Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, People's Republic of China; Division of Molecular Pharmacology of Infectious Agents, Department of Molecular Microbiology and Immunology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki City, Nagasaki Prefecture 852-8521, Japan; and Department of Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA.
authors.forename: Bo
authors.initials: B
authors.lastname: Han
authors.affiliation: Institute of Biomedicine & National Engineering Research Center of Genetic Medicine, Department of Cellular Biology, Jinan University, Guangzhou 510632, Guangdong, People's Republic of China; Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Centre for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China; Department of Pharmacy, College of Food and Pharmacy & Medical, Zhejiang Ocean University, Zhoushan 316002, People's Republic of China; Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, People's Republic of China; Division of Molecular Pharmacology of Infectious Agents, Department of Molecular Microbiology and Immunology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki City, Nagasaki Prefecture 852-8521, Japan; and Department of Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA.
authors.forename: Zhou
authors.initials: Z
authors.lastname: Peng
authors.affiliation: Institute of Biomedicine & National Engineering Research Center of Genetic Medicine, Department of Cellular Biology, Jinan University, Guangzhou 510632, Guangdong, People's Republic of China; Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Centre for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China; Department of Pharmacy, College of Food and Pharmacy & Medical, Zhejiang Ocean University, Zhoushan 316002, People's Republic of China; Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, People's Republic of China; Division of Molecular Pharmacology of Infectious Agents, Department of Molecular Microbiology and Immunology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki City, Nagasaki Prefecture 852-8521, Japan; and Department of Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA Institute of Biomedicine & National Engineering Research Center of Genetic Medicine, Department of Cellular Biology, Jinan University, Guangzhou 510632, Guangdong, People's Republic of China; Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Centre for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China; Department of Pharmacy, College of Food and Pharmacy & Medical, Zhejiang Ocean University, Zhoushan 316002, People's Republic of China; Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, People's Republic of China; Division of Molecular Pharmacology of Infectious Agents, Department of Molecular Microbiology and Immunology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki City, Nagasaki Prefecture 852-8521, Japan; and Department of Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA.
authors.forename: Feng
authors.initials: F
authors.lastname: Ge
authors.affiliation: Institute of Biomedicine & National Engineering Research Center of Genetic Medicine, Department of Cellular Biology, Jinan University, Guangzhou 510632, Guangdong, People's Republic of China; Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Centre for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China; Department of Pharmacy, College of Food and Pharmacy & Medical, Zhejiang Ocean University, Zhoushan 316002, People's Republic of China; Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, People's Republic of China; Division of Molecular Pharmacology of Infectious Agents, Department of Molecular Microbiology and Immunology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki City, Nagasaki Prefecture 852-8521, Japan; and Department of Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA.
authors.forename: Bo
authors.initials: B
authors.lastname: Wei
authors.affiliation: Institute of Biomedicine & National Engineering Research Center of Genetic Medicine, Department of Cellular Biology, Jinan University, Guangzhou 510632, Guangdong, People's Republic of China; Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Centre for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China; Department of Pharmacy, College of Food and Pharmacy & Medical, Zhejiang Ocean University, Zhoushan 316002, People's Republic of China; Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, People's Republic of China; Division of Molecular Pharmacology of Infectious Agents, Department of Molecular Microbiology and Immunology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki City, Nagasaki Prefecture 852-8521, Japan; and Department of Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA.
authors.forename: Mingxian
authors.initials: M
authors.lastname: Liu
authors.affiliation: Institute of Biomedicine & National Engineering Research Center of Genetic Medicine, Department of Cellular Biology, Jinan University, Guangzhou 510632, Guangdong, People's Republic of China; Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Centre for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China; Department of Pharmacy, College of Food and Pharmacy & Medical, Zhejiang Ocean University, Zhoushan 316002, People's Republic of China; Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, People's Republic of China; Division of Molecular Pharmacology of Infectious Agents, Department of Molecular Microbiology and Immunology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki City, Nagasaki Prefecture 852-8521, Japan; and Department of Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA.
authors.forename: Meiying
authors.initials: M
authors.lastname: Zhang
authors.affiliation: Institute of Biomedicine & National Engineering Research Center of Genetic Medicine, Department of Cellular Biology, Jinan University, Guangzhou 510632, Guangdong, People's Republic of China; Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Centre for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China; Department of Pharmacy, College of Food and Pharmacy & Medical, Zhejiang Ocean University, Zhoushan 316002, People's Republic of China; Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, People's Republic of China; Division of Molecular Pharmacology of Infectious Agents, Department of Molecular Microbiology and Immunology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki City, Nagasaki Prefecture 852-8521, Japan; and Department of Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA.
authors.forename: Chuiwen
authors.initials: C
authors.lastname: Qian
authors.affiliation: Institute of Biomedicine & National Engineering Research Center of Genetic Medicine, Department of Cellular Biology, Jinan University, Guangzhou 510632, Guangdong, People's Republic of China; Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Centre for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China; Department of Pharmacy, College of Food and Pharmacy & Medical, Zhejiang Ocean University, Zhoushan 316002, People's Republic of China; Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, People's Republic of China; Division of Molecular Pharmacology of Infectious Agents, Department of Molecular Microbiology and Immunology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki City, Nagasaki Prefecture 852-8521, Japan; and Department of Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA.
authors.forename: Zhibo
authors.initials: Z
authors.lastname: Hou
authors.affiliation: Institute of Biomedicine & National Engineering Research Center of Genetic Medicine, Department of Cellular Biology, Jinan University, Guangzhou 510632, Guangdong, People's Republic of China; Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Centre for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China; Department of Pharmacy, College of Food and Pharmacy & Medical, Zhejiang Ocean University, Zhoushan 316002, People's Republic of China; Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, People's Republic of China; Division of Molecular Pharmacology of Infectious Agents, Department of Molecular Microbiology and Immunology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki City, Nagasaki Prefecture 852-8521, Japan; and Department of Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA.
authors.forename: Ge
authors.initials: G
authors.lastname: Liu
authors.affiliation: Institute of Biomedicine & National Engineering Research Center of Genetic Medicine, Department of Cellular Biology, Jinan University, Guangzhou 510632, Guangdong, People's Republic of China; Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Centre for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China; Department of Pharmacy, College of Food and Pharmacy & Medical, Zhejiang Ocean University, Zhoushan 316002, People's Republic of China; Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, People's Republic of China; Division of Molecular Pharmacology of Infectious Agents, Department of Molecular Microbiology and Immunology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki City, Nagasaki Prefecture 852-8521, Japan; and Department of Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA.
authors.forename: Chaowan
authors.initials: C
authors.lastname: Guo
authors.affiliation: Institute of Biomedicine & National Engineering Research Center of Genetic Medicine, Department of Cellular Biology, Jinan University, Guangzhou 510632, Guangdong, People's Republic of China; Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Centre for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China; Department of Pharmacy, College of Food and Pharmacy & Medical, Zhejiang Ocean University, Zhoushan 316002, People's Republic of China; Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, People's Republic of China; Division of Molecular Pharmacology of Infectious Agents, Department of Molecular Microbiology and Immunology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki City, Nagasaki Prefecture 852-8521, Japan; and Department of Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA.
authors.forename: Yifei
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Institute of Biomedicine & National Engineering Research Center of Genetic Medicine, Department of Cellular Biology, Jinan University, Guangzhou 510632, Guangdong, People's Republic of China; Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Centre for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China; Department of Pharmacy, College of Food and Pharmacy & Medical, Zhejiang Ocean University, Zhoushan 316002, People's Republic of China; Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, People's Republic of China; Division of Molecular Pharmacology of Infectious Agents, Department of Molecular Microbiology and Immunology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki City, Nagasaki Prefecture 852-8521, Japan; and Department of Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA.
authors.forename: Kaio
authors.initials: K
authors.lastname: Kitazato
authors.affiliation: Institute of Biomedicine & National Engineering Research Center of Genetic Medicine, Department of Cellular Biology, Jinan University, Guangzhou 510632, Guangdong, People's Republic of China; Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Centre for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China; Department of Pharmacy, College of Food and Pharmacy & Medical, Zhejiang Ocean University, Zhoushan 316002, People's Republic of China; Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, People's Republic of China; Division of Molecular Pharmacology of Infectious Agents, Department of Molecular Microbiology and Immunology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki City, Nagasaki Prefecture 852-8521, Japan; and Department of Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA.
authors.forename: Guoying
authors.initials: G
authors.lastname: Yu
authors.affiliation: Institute of Biomedicine & National Engineering Research Center of Genetic Medicine, Department of Cellular Biology, Jinan University, Guangzhou 510632, Guangdong, People's Republic of China; Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Centre for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China; Department of Pharmacy, College of Food and Pharmacy & Medical, Zhejiang Ocean University, Zhoushan 316002, People's Republic of China; Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, People's Republic of China; Division of Molecular Pharmacology of Infectious Agents, Department of Molecular Microbiology and Immunology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki City, Nagasaki Prefecture 852-8521, Japan; and Department of Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA.
authors.forename: Chunbin
authors.initials: C
authors.lastname: Zou
authors.affiliation: Institute of Biomedicine & National Engineering Research Center of Genetic Medicine, Department of Cellular Biology, Jinan University, Guangzhou 510632, Guangdong, People's Republic of China; Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Centre for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China; Department of Pharmacy, College of Food and Pharmacy & Medical, Zhejiang Ocean University, Zhoushan 316002, People's Republic of China; Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, People's Republic of China; Division of Molecular Pharmacology of Infectious Agents, Department of Molecular Microbiology and Immunology, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki City, Nagasaki Prefecture 852-8521, Japan; and Department of Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA xiongsheng@jnu.edu.cn.
authors.forename: Sheng
authors.initials: S
authors.lastname: Xiong
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000737:chemistry / Q000494:pharmacology*; D002352:Carrier Proteins / Q000737:chemistry / Q000494:pharmacology*; D002642:Chick Embryo; D004285:Dogs; D005260:Female; D066298:In Vitro Techniques; D009980:Influenza A virus / Q000187:drug effects* / Q000502:physiology; D061985:Madin Darby Canine Kidney Cells; D051379:Mice; D008807:Mice, Inbred BALB C; D008826:Microbial Sensitivity Tests; D011092:Polyethylene Glycols / Q000737:chemistry*; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000998:Antiviral Agents; D001426:Bacterial Proteins; D002352:Carrier Proteins; C107733:cyanovirin N; D011092:Polyethylene Glycols
keywords: Cyanovirin-N; PEGylation; influenza A virus; mice; polypeptide linker