Virus-like particles and methods of use

Patent metadata

Field	Value
Publication number	US-2017073377-A1
Application number	US-201615279592-A
Country	US
Kind code	A1
Filing date	Sep 29, 2016
Priority date	Jan 31, 2011
Publication date	Mar 16, 2017
Grant date	—

Abstract

The invention features modified alphavirus or flavivirus virus-like particles (VLPs). The invention provides methods, compositions, and kits featuring the modified VLPs. The invention also features methods for enhancing production of modified VLPs for use in the prevention or treatment of alphavirus and flavivirus-mediated diseases. The invention also provides methods for delivering agents to a cell using the modified VLPs.

First claim

1 - 103 . (canceled)

104 . An isolated polynucleotide encoding an altered viral protein selected from the group consisting of:
a. an alphavirus or flavivirus E2 protein comprising at least one alteration, relative to the wild-type amino acid sequence, at one or more amino acid locations corresponding to at least one amino acid position selected from the group consisting of H170, K200, K233, K234, R251, and H256 of Chikungunya E2 protein; and b. an alphavirus or flavivirus capsid protein comprising at least one alteration, relative to the wild type sequence, in the Nuclear Localization Signal (NLS); wherein the altered protein is capable of self-assembling into a virus like particle (VLP); and, wherein the at least one alteration enhances production of VLPs.

105 . The isolated polynucleotide of claim 104 , wherein the alphavirus or flavivirus is selected from the group consisting of Eastern equine encephalitis virus (EEEV), Western equine encephalitis virus (WEEV), Venezuelan equine encephalitis virus (VEEV), Semliki Forest virus (SFV), Chikungunya virus (CHIKV), O'nyong-nyong virus, Sindbis virus, Mayaro virus, Ross River virus, Barmah Forest virus, Ockelbo virus, Yellow Fever Virus (YFV), Dengue Virus (DENV), Japanese Encephalitis Virus (JEV), Tick-Borne Encephalitis Virus (TBEV), and West Nile Virus (WNV).

106 . The isolated polynucleotide of claim 104 , wherein the at least one alteration in the NLS is in at least one amino acid position selected from the group consisting of:
a. amino acids 67-70 of an EEEV capsid protein; b. amino acids 67-70 of an WEEV capsid protein; c. amino acids 64-68 of an VEEV capsid protein; d. amino acids 62-69 of a CHIKV capsid protein; e. amino acids 71-74 of a Ross River virus capsid protein; and f. amino acids 64-68 of a Barmah Forest virus capsid protein.

107 . The isolated polynucleotide of claim 104 , wherein the at least one alteration comprises a substitution in a charged amino acid of the NLS.

108 . A kit comprising the isolated polynucleotide of claim 104 .

109 . A method for producing a VLP, comprising introducing into a cell a polynucleotide encoding an altered viral protein selected from the group consisting of:
a. an alphavirus or flavivirus E2 protein comprising at least one alteration, relative to the wild-type amino acid sequence, at one or more amino acid locations corresponding to at least one amino acid position selected from the group consisting of H170, K200, K233, K234, R251, and H256 of Chikungunya E2 protein; and b. an alphavirus or flavivirus capsid protein comprising at least one alteration, relative to the wild type sequence, in the Nuclear Localization Signal (NLS); wherein the altered protein is capable of self-assembling into a virus like particle (VLP); and, wherein the at least one alteration enhances production of VLPs.

110 . The method of claim 109 , wherein the alphavirus or flavivirus is selected from the group consisting of Eastern equine encephalitis virus (EEEV), Western equine encephalitis virus (WEEV), Venezuelan equine encephalitis virus (VEEV), Semliki Forest virus (SFV), Chikungunya virus (CHIKV), O'nyong-nyong virus, Sindbis virus, Mayaro virus, Ross River virus, Barmah Forest virus, Ockelbo virus, Yellow Fever Virus (YFV), Dengue Virus (DENV), Japanese Encephalitis Virus (JEV), Tick-Borne Encephalitis Virus (TBEV), and West Nile Virus (WNV).

111 . The method of claim 109 , wherein the at least one alteration in the NLS is in at least one amino acid position selected from the group consisting of:
a. amino acids 67-70 of an EEEV capsid protein; b. amino acids 67-70 of an WEEV capsid protein; c. amino acids 64-68 of an VEEV capsid protein; d. amino acids 62-69 of a CHIKV capsid protein; e. amino acids 71-74 of a Ross River virus capsid protein; and f. amino acids 64-68 of a Barmah Forest virus capsid protein.

112 . The method of claim 109 , wherein the at least one alteration comprises a substitution in a charged amino acid of the NLS.

Assignees

• The Usa As Represented By The Secretary Dept Of Health And Human Services

Inventors

• Nabel Gary J
• Rao Srinivas
• Akahata Wataru

Classifications

• A61K39/12 Primary

  Viral antigens · CPC title

• C12N7/00

  Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00) · CPC title

• C12N2770/36122

  New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes · CPC title

• A61K2039/5258

  Virus-like particles · CPC title

• C12N2770/36151

  Methods of production or purification of viral material · CPC title