# US 10,766,930 B2 · Fusion proteins and use thereof for preparing vaccines > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,766,930 B2 - Document ID: US10766930B2 - Status: Granted - Kind: B2 - Application number: 16/310,666 - Filed: 2017-06-16 - Published: 2019-12-26 - Granted: 2020-09-08 - Earliest claimed: 2016-06-17 - Assignees: Université De Tours; Centre Hospitalier Regional Universitaire De Tours; Institut National De La Sante Et De La Recherche Medicale (Inserm) - Applicants: Universite De Tours; Centre Hospitalier Regional Universitaire De Tours; Institut National De La Sante Et De La Recherche Medicale-Inserm - Inventors: Christophe Hourioux; Romuald Patient; Elodie Beaumont; Philippe Roingeard - Web page: https://patentgrep.com/patent/US10766930B2 ## CPC - C07K 14/005: from viruses - A61K 39/12: Viral antigens - A61K 39/292: {Serum hepatitis virus, hepatitis B virus, e.g. Australia antigen} - A61P 31/12: Antivirals - A61P 31/14: for RNA viruses - A61P 31/20: for DNA viruses - C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00) - C12N 15/62: DNA sequences coding for fusion proteins - C12P 21/02: having a known sequence of two or more amino acids, e.g. glutathione - A61K 2039/5258: {Virus-like particles} - A61K 2039/70: {Multivalent vaccine} - C07K 2319/03: containing a transmembrane segment - C07K 2319/33: fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies - C12N 2730/10134: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein - C12N 2770/24134: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein - Y02A 50/30: Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change - 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) - 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) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)} - Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE ## Abstract An immunogenic fusion protein includes at least the following two peptides a) —on the C-terminal side, a first peptide constituted of: —the amino acid sequence of the protein S or the protein M of a human hepatitis B virus (HBV) isolate, which protein S or protein M is optionally deleted at the N-terminal end thereof, and b) —on the N-terminal side, a second peptide constituted of: —the sequence of amino acids of at least one transmembrane domain and the ectodomain of at least one protein of a Zika virus isolate selected from the envelope protein E or a fusion peptide including the envelope protein E and the protein prM. ## Claims ### Claim 1 An immunogenic fusion protein comprising at least two peptides: a) on the C-terminal side, a first peptide which consists of: an amino acid sequence of the protein S or the protein M of a human hepatitis B virus (HBV) isolate, wherein the sequence of the protein S is chosen from the group consisting of SEQ ID NO: 1 and 2, and the sequence of the protein M is chosen from the group consisting of SEQ ID NO: 3 and 4, or an amino acid sequence with a percent identity of at least 95% with said amino acid sequence of the protein S or the protein M, provided that said amino acid sequence maintains the ability to form subviral, non-infectious particles and/or immunogenic properties against the HBV virus, or, and b) on the N-terminal side, a second peptide which consists of: a sequence of amino acids comprising at least one transmembrane domain and the ectodomain of at least one protein of a Zika virus isolate, or an amino acid sequence with a percent identity of at least 95% with said amino acid sequence of at least one transmembrane domain and the ectodomain of at least one protein of a Zika virus isolate, provided that said amino acid sequence maintains the ability to form subviral, non-infectious particles and/or immunogenic properties against the Zika virus, said protein of a Zika virus isolate being chosen from among the envelope protein E represented by SEQ ID NO. 15 or a fusion peptide comprising the envelope protein E and the protein prM represented by SEQ ID NO. 50. ### Claim 2 (depends on 1) An immunogenic fusion protein according to claim 1, wherein the first peptide located on the C-terminal side thereof consists of: the amino acid sequence of the protein S of a human HBV isolate, wherein the sequence of the protein S is represented by of SEQ ID NO: 1, or an amino acid sequence presenting a percent identity of at least 95% with said amino acid sequence of the protein S represented by of SEQ ID NO: 1, provided that said amino acid sequence maintains the ability to form subviral, non-infectious particles and/or the immunogenic properties against the human HBV. ### Claim 3 (depends on 1) An immunogenic fusion protein according to claim 1, wherein the first peptide located on the C-terminal side thereof consists of: the amino acid sequence of the protein S of a human HBV isolate, wherein the sequence of the protein S is represented by of SEQ ID NO: 2, or an amino acid sequence presenting a percent identity of at least 95% with said amino acid sequence of the protein S represented by of SEQ ID NO: 2, provided that said amino acid sequence maintains the ability to form subviral, non-infectious particles and/or the immunogenic properties against the human HBV. ### Claim 4 (depends on 1) An immunogenic fusion protein according to claim 1, wherein the first peptide located on the C-terminal side thereof consists of: the amino acid sequence of the protein M of a human HBV isolate, wherein the sequence of the protein M is represented by of SEQ ID NO: 3, or an amino acid sequence presenting a percent identity of at least 95% with said amino acid sequence of the protein M deleted from a sequence of 1 to 54 amino acids located at the N-terminal end thereof, provided that said amino acid sequence maintains the ability to form subviral, non-infectious particles and/or the immunogenic properties against the human HBV. ### Claim 5 (depends on 1) An immunogenic fusion protein according to claim 1, wherein the first peptide located on the C-terminal side thereof consists of: the amino acid sequence of the protein M of a human HBV isolate, wherein the sequence of the protein M is represented by of SEQ ID NO: 4, or an amino acid sequence presenting a percent identity of at least 95% with said amino acid sequence of the protein M not deleted from a sequence of 1 to 54 amino acids located at the N-terminal end thereof, provided that said amino acid sequence maintains the ability to form subviral, non-infectious particles and/or the immunogenic properties against the human HBV. ### Claim 6 (depends on 1) An immunogenic fusion protein according to claim 1, wherein the second peptide located on the N-terminal side thereof consists of: the amino acid sequence of at least one transmembrane domain and the ectodomain of the envelope protein E of a Zika virus isolate, or an amino acid sequence presenting an identity percent of at least 95%, with said amino acid sequence of at least one transmembrane domain and the ectodomain of the envelope protein E of a Zika virus isolate, provided that said amino acid sequence maintains the ability to form subviral, non-infectious particles and/or the immunogenic properties against the Zika virus. ### Claim 7 (depends on 1) An immunogenic fusion protein according to claim 1, wherein the second peptide located on the N-terminal side thereof consists of: the amino acid sequence of a fusion peptide comprising at least one transmembrane domain and the ectodomain of the envelope protein E of a Zika virus isolate and the protein prM of a Zika virus isolate, wherein said amino acid sequence is chosen from among the envelope protein E represented by SEQ ID NO. 15 or a fusion peptide comprising the envelope protein E and the protein prM represented by SEQ ID NO. 50, or an amino acid sequence presenting an identity percent of at 95%, with said amino acid sequence of said fusion peptide, provided that said amino acid sequence maintains the ability to form subviral, non-infectious particles and/or the immunogenic properties against the Zika virus. ### Claim 8 (depends on 1) An immunogenic fusion protein according to claim 1, wherein said fusion protein comprises: the amino acid sequence represented by: SEQ ID NO. 7 or SEQ ID NO. 17, or SEQ ID NO. 8 or SEQ ID NO. 18, or SEQ ID NO. 9 or SEQ ID NO. 19, or SEQ ID NO. 10 or SEQ ID NO. 20, or SEQ ID NO. 11 or SEQ ID NO. 21, or SEQ ID NO. 12 or SEQ ID NO. 22, or SEQ ID NO. 13 or SEQ ID NO. 23, or SEQ ID NO. 14 or SEQ ID NO. 24, or an amino acid sequence presenting an identity percent of at least 95% with said SEQ ID NO. 7, SEQ ID NO. 17, SEQ ID NO. 8, SEQ ID NO. 18, SEQ ID NO. 9, SEQ ID NO. 19, SEQ ID NO. 10, SEQ ID NO. 20, SEQ ID NO. 11, SEQ ID NO. 21, SEQ ID NO. 12, SEQ ID NO. 22, SEQ ID NO. 13, SEQ ID NO. 23, SEQ ID NO. 14, SEQ ID NO. 24, provided that said amino acid sequence maintains the ability to form subviral, non-infectious particles and/or the immunogenic properties against the human HBV and/or the Zika virus, or the amino acid sequence of a synthetic variant derived from said SEQ ID NO. 7, SEQ ID NO. 17, SEQ ID NO. 8, SEQ ID NO. 18, SEQ ID NO. 9, SEQ ID NO. 19, SEQ ID NO. 10, SEQ ID NO. 20, SEQ ID NO. 11, SEQ ID NO. 21, SEQ ID NO. 12, SEQ ID NO. 22, SEQ ID NO. 13, SEQ ID NO. 23, SEQ ID NO. 14, SEQ ID NO. 24, provided that said amino acid sequence maintains the ability to form subviral, non-infectious particles and/or the immunogenic properties against the human HBV and/or the Zika virus. ### Claim 9 (depends on 1) Nucleic acid molecules coding a fusion protein according to claim 1. ### Claim 10 (depends on 1) A subviral, non-infectious and immunogenic particle comprising the following proteins: a protein comprising the wild-type domain S of the surface antigen of a hepatitis B virus isolate, and the fusion protein according to claim 1. ### Claim 11 (depends on 1) An immunogenic fusion protein according to claim 1, for its use as a medication, notably as a vaccine. ### Claim 12 (depends on 1) An immunogenic fusion protein according to claim 1, for its use in preventing and/or treating hepatitis B and/or Zika virus infections. ### Claim 13 (depends on 1) An immunogenic fusion protein according to claim 1, wherein the second peptide located on the N-terminal side thereof consists of: the sequence of amino acids of at least one transmembrane domain chosen from the group consisting of the amino acid sequences between positions 456-484 of SEQ ID NO: 15, 485-504 of SEQ ID NO: 15, 124-143 of SEQ ID NO: 50, and 150-164 of SEQ ID NO: 50, and the amino acid sequence of the ectodomain chosen from the group consisting of the amino acid sequences between positions 1-455 of SEQ ID NO: 15 and 1-123 of SEQ ID NO: 50 of at least one protein of a Zika virus isolate, or an amino acid sequence with a percent identity of at least 95% with said amino acid sequence of at least one transmembrane domain and the ectodomain of at least one protein of a Zika virus isolate, provided that said amino acid sequence maintains the ability to form subviral, non-infectious particles and/or immunogenic properties against the Zika virus, said protein of a Zika virus isolate being chosen from among the envelope protein E represented by SEQ ID NO. 15 and a fusion peptide comprising the envelope protein E and the protein prM represented by SEQ ID NO. 50.