# US 9,434,959 B2 · Virus-like particles for treatment of viral infections > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 9,434,959 B2 - Document ID: US9434959B2 - Status: Granted - Kind: B2 - Application number: 14/249,864 - Filed: 2014-04-10 - Published: 2014-07-31 - Granted: 2016-09-06 - Earliest claimed: 2006-06-15 - Assignees: Ben-Gurion University of the Negev Research and Development Authority - Applicants: Ben Gurion University of the Negev Research and Development Authority - Inventors: Leslie Lobel; Guy Gubi - Web page: https://patentgrep.com/patent/US9434959B2 ## CPC - C12N 15/86: Viral vectors - A61K 39/12: Viral antigens - A61K 39/145: Orthomyxoviridae, e.g. influenza virus - C07K 14/005: from viruses - C07K 16/108: Orthomyxoviridae (F), e.g. influenza virus - C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00) - A61K 2039/5258: {Virus-like particles} - C07H 21/04: with deoxyribosyl as saccharide radical - C07K 2317/622: Single chain antibody (scFv) - C07K 2317/80: remaining in the (producing) cell, i.e. intracellular antibodies or intrabodies - C12N 7/02: Recovery or purification - C12N 7/045: {Pseudoviral particles; Non infectious pseudovirions, e.g. genetically engineered} - C12N 2760/14134: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein - C12N 2760/16122: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes - C12N 2760/16123: Virus like particles [VLP] - C12N 2760/16143: viral genome or elements thereof as genetic vector - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - C07H: SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS (derivatives of aldonic or saccharic acids C07C, C07D; aldonic acids, saccharic acids C07C59/105, C07C59/285; cyanohydrins C07C255/16; glycals C07D; compounds of unknown constitution C07G; polysaccharides, derivatives thereof C08B; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; sugar industry C13) ## Abstract The invention provides virus-like particles for treatment of viral infections based on the virus causing the infection. The virus-like particles comprise the virus recombinant proteins that form a capsid, recombinant virus membrane proteins attached to the capsid and vRNA packaged within said capsid. The vRNA is generated from a DNA sequence encoding a polypeptide capable of specifically binding to a constant region of a nonstructural protein of the virus that is essential for propagation of the virus. ## Claims ### Claim 1 A virus-like particle adapted for treating a viral infection, wherein said virus-like particle is based on an infecting virus causing the viral infection, with the proviso that said infecting virus is not a hepatitis C virus, and said virus-like particle comprises: recombinant viral proteins which are of the same species as said infecting virus and form a capsid, recombinant viral membrane proteins which are of the same species as said infecting virus and are attached to the surface of said capsid, and vRNA packaged within said capsid, wherein said vRNA is generated by intracellular transcription of a DNA sequence encoding a polypeptide capable of specifically binding to a constant region of a nonstructural protein of said infecting virus, said nonstructural protein being essential for propagation of said infecting virus, whereby said binding interferes with the activity of said nonstructural viral protein and inhibits the propagation of said virus. ### Claim 2 (depends on 1) The virus-like particle according to claim 1, wherein said infecting virus is selected from the group consisting of an RNA negative sense virus, an RNA ambisense virus, and an RNA positive sense virus. ### Claim 3 (depends on 2) The virus-like particle according to claim 2, wherein said negative sense virus is selected from the group consisting of orthomyxoviridae, paramyxoviridae, filoviridae, rhabdoviridae, arenaviridae and bunyaviridae families of virus and said positive sense virus is a flaviviridae family of virus. ### Claim 4 (depends on 3) The virus-like particle according to claim 3, wherein said orthomyxoviridae virus is an influenza virus, said paramyxoviridae virus is human respiratory syncytial virus, said filoviridae virus is ebola virus or Marburg virus, said rhabdoviridae virus is rabies virus, said arenaviridae virus is lassa virus, said bunyaviridae virus is hanta virus. ### Claim 5 (depends on 4) The virus-like particle according to claim 4, wherein said virus is influenza A virus. ### Claim 6 (depends on 1) The virus-like particle according to claim 1, wherein said virus nonstructural protein is selected from the group consisting of: the PB1, PB2, or PA subunit of the RNA dependent RNA polymerase (RDRP), the nucleoprotein (NP) or the M protein of an influenza virus; the NP or RDRP of a human respiratory syncytial virus; the L, VP35, NP, or VP30 protein of an ebola or Marburg virus; the NP or RDRP protein of a rabies virus; the L (RDRP) or N (Nucleoprotein) protein of a lassa virus; and the L (RDRP)or N (Nucleoprotein) protein of a hanta virus. ### Claim 7 (depends on 6) The virus-like particle according to claim 6, wherein said nonstructural protein is selected from the group consisting of the PB1, PB2, and PA subunits of the RNA dependent RNA polymerase (RDRP), the nucleoprotein (NP), and the M protein of influenza virus. ### Claim 8 (depends on 6) The virus-like particle according to claim 6, wherein said virus nonstructural protein is involved in a cellular process essential for propagation of the virus selected from the group consisting of replication of the viral genetic material, packaging of said genetic material into the viral capsid and budding of the capsid from the host cell. ### Claim 9 (depends on 8) The virus-like particle according to claim 8, wherein said virus is influenza A virus, said cellular process is replication of the viral genetic material, and said nonstructural viral protein is the PB1 subunit of influenza A virus RDRP. ### Claim 10 (depends on 9) The virus-like particle according to claim 9, wherein said DNA sequence encodes (i) a single chain variable fragment antibody (ScFv) polypeptide capable of specifically binding to the PA binding domain of the PB1 subunit of the influenza A virus RDRP, or (ii) a peptide fragment comprising the N-terminal amino acid sequence of the PB1 subunit, which peptide fragment is capable of binding to the PB1 binding domain of the PA subunit so as to compete with the PB1 subunit of influenza A virus RDRP for binding to the PA subunit and inhibit viral propagation. ### Claim 11 (depends on 10) The virus-like particle according to claim 10, wherein said DNA sequence encodes a scFv polypeptide. ### Claim 12 (depends on 11) The virus-like particle according to claim 11, wherein said scFv polypeptide specifically binds to the PB1 N-terminus amino acid sequence of SEQ ID NO: 1. ### Claim 13 (depends on 12) The virus-like particle according to claim 12, wherein said scFv is encoded by a DNA sequence represented by SEQ ID NO: 2. ### Claim 14 (depends on 10) The virus-like particle according to claim 10, wherein said peptide fragment is represented by SEQ ID NO: 1. ### Claim 15 (depends on 1) The virus-like particle according to claim 1, wherein said recombinant virus membrane proteins are hemagglutinin and neuraminidase, and said vRNA is generated by intracellular transcription of the DNA sequence of an scFv. ### Claim 16 (depends on 1) A method for producing a virus-like particle according to claim 1, comprising introducing into an eukaryotic host cell: (i) a plasmid comprising a DNA sequence encoding a polypeptide capable of specifically binding to a constant region of a virus nonstructural protein that is essential for propagation of said infecting virus, whereby the binding interferes with the activity of said nonstructural viral protein and inhibits the propagation of said virus, wherein said DNA sequence is operably linked to a promoter selected from the group consisting of an RNA polymerase I promoter, a T3 RNA polymerase promoter and a T7 RNA polymerase promoter and is flanked by a terminator selected from the group consisting of an RNA polymerase I transcription terminator, a T3 RNA polymerase transcription terminator and a T7 RNA transcription terminator sequence and by viral transcription and packaging signal sequences; (ii) one or more plasmids comprising DNA sequences encoding said virus proteins that form a capsid; and (iii) one or more plasmids comprising DNA sequences encoding said virus membrane proteins; whereby the virus proteins expressed by said plasmids (ii) in said host cell form a capsid, and the intracellular transcription of the DNA of plasmid (i) generates a vRNA that is packaged within said capsid, which during the budding process acquires cell membrane lipids along with the virus membrane proteins expressed by plasmids (iii), thus producing said virus-like particle, which are released from the host cell. ### Claim 17 (depends on 1) A pharmaceutical composition comprising the virus-like particle of claim 1 and a pharmaceutically acceptable carrier. ### Claim 18 (depends on 1) A virus-like particle according to claim 1, wherein said vRNA contains a sequence encoding a polypeptide capable of specifically binding to a constant region of a nonstructural protein of said infecting virus instead of containing the genetic material of the wild-type virus.