WO2023085956A1 - Nouveaux vaccins thérapeutiques - Google Patents

Nouveaux vaccins thérapeutiques Download PDF

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Publication number
WO2023085956A1
WO2023085956A1 PCT/NZ2022/050141 NZ2022050141W WO2023085956A1 WO 2023085956 A1 WO2023085956 A1 WO 2023085956A1 NZ 2022050141 W NZ2022050141 W NZ 2022050141W WO 2023085956 A1 WO2023085956 A1 WO 2023085956A1
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seq
immunogenic peptide
cluster
peptide defined
hbv
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PCT/NZ2022/050141
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English (en)
Inventor
Alessandro Sette
John Sidney
Francis Chisari
Shivali GULAB
Gavin Painter
Ian Hermans
John Steemson
Peter Dunbar
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Avalia Immunotherapies Limited
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Publication of WO2023085956A1 publication Critical patent/WO2023085956A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/20—Antivirals for DNA viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/12—Viral antigens
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/515—Animal cells
    • A61K2039/5154—Antigen presenting cells [APCs], e.g. dendritic cells or macrophages
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/70—Multivalent vaccine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/46—Cellular immunotherapy
    • A61K39/462—Cellular immunotherapy characterized by the effect or the function of the cells
    • A61K39/4622—Antigen presenting cells
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2730/00—Reverse transcribing DNA viruses
    • C12N2730/00011—Details
    • C12N2730/10011—Hepadnaviridae
    • C12N2730/10111—Orthohepadnavirus, e.g. hepatitis B virus
    • C12N2730/10122—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2730/00—Reverse transcribing DNA viruses
    • C12N2730/00011—Details
    • C12N2730/10011—Hepadnaviridae
    • C12N2730/10111—Orthohepadnavirus, e.g. hepatitis B virus
    • C12N2730/10134—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein

Definitions

  • the present invention is concerned with novel therapeutic vaccines for infectious disease.
  • the present invention provides immunogenic peptides, vaccine compositions and methods of therapy targeting chronic hepatitis B virus (HBV) infection in humans.
  • the immunogenic peptides include a unique selection/combinatorial assembly of human leukocyte antigen (HLA) class I restricted T-cell epitopes derived from hepatitis B virus antigens to yield vaccine compositions that achieve extensive population coverage, for example, by geography and/or ethnicity.
  • HLA human leukocyte antigen
  • Hepatitis B is a potentially life-threatening liver infection caused by the hepatitis B virus (HBV). It is a major and growing global health problem and significant cause of morbidity and mortality worldwide.
  • HBV hepatitis B virus
  • HBV hepatitis B surface antigen positive
  • HBV is most prevalent in the Western Pacific Region and the African Region, where 6.2% and 6.1% of the adult population is infected, respectively. In highly endemic areas, HBV is commonly transmitted from mother to child at birth (perinatal transmission), or through horizontal transmission (by needlestick injury, tattooing, piercing and exposure to infected blood and body fluids, such as saliva, vaginal, and seminal fluids).
  • Hepatitis B can be prevented by vaccination, and vaccination campaigns for HBV are central to WHO's ambition to eradicate HBV globally.
  • the oral treatments tenofovir or entecavir are considered the most potent drugs to suppress HBV. Compared with other drugs, they rarely lead to drug resistance, are simple to take, and require only limited monitoring. Treatment can slow the progression of cirrhosis, reduce incidence of liver cancer and improve long term survival.
  • the required life-long therapy is associated wide side-effects and high cost, limiting patient access in low-income settings.
  • the present invention seeks to address these limitations by providing novel therapeutic vaccines for the treatment of chronic hepatitis B virus infection.
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three, at least four, at least five or at least six hepatitis B virus (HBV) epitopes selected from FLPSDFFPSV (SEQ ID NO: 12), LPSDFFPSV (SEQ ID NO: 13), DFFPSIRDL (SEQ ID NO: 14), FFPSIRDLL (SEQ ID NO: 15), DLLDTASALY (SEQ ID NO: 16) and LLDTASALY (SEQ ID NO: 17), or a variant sequence which shares at least 75%, 80%, 85%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 12-17.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising at least three, at least four, at least five or at least six hepatitis B virus (HBV) epitopes selected from FLPSDFFPSV (SEQ ID NO: 12), LPSDFFPSV (SEQ ID NO: 13), DFFPSIRDL (SEQ ID NO: 14), FFPSIRDLL (SEQ ID NO: 15), DLLDTASALY (SEQ ID NO: 16) and LLDTASALY (SEQ ID NO: 17), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 12- 17.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising three, four, five or six hepatitis B virus (HBV) epitopes selected from FLPSDFFPSV (SEQ ID NO: 12), LPSDFFPSV (SEQ ID NO: 13), DFFPSIRDL (SEQ ID NO: 14), FFPSIRDLL (SEQ ID NO: 15), DLLDTASALY (SEQ ID NO: 16) and LLDTASALY (SEQ ID NO: 17), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 12-17, provided that the immunogenic peptide or immunogenic peptide group comprises at least one epitope restricted by human leukocyte antigen (HLA) A*02:01 and/or HLA A*24:02.
  • HLA human leukocyte antigen
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising three, four, five or six hepatitis B virus (HBV) epitopes selected from SEQ ID Nos: FLPSDFFPSV (SEQ ID NO: 12), LPSDFFPSV (SEQ ID NO: 13), DFFPSIRDL (SEQ ID NO: 14), FFPSIRDLL (SEQ ID NO: 15), DLLDTASALY (SEQ ID NO: 16) and LLDTASALY (SEQ ID NO: 17), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 12-17, provided that the immunogenic peptide or immunogenic peptide group comprises FLPSDFFPSV (SEQ ID NO: 12).
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence LSFLPSDFFPSVRDLLDTASALY (Cluster 1; SEQ ID NO: 1) or FLPSDFFPSVRDLLDTASALY (Cluster IB; SEQ ID NO: 10), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 1 or SEQ ID NO: 10, respectively.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three, at least four, at least five or at least six hepatitis B virus (HBV) epitopes selected from YVNTNMGLK (SEQ ID NO: 18), GLKILQLL (SEQ ID NO: 19), LWFHISCLTF (SEQ ID NO: 20), HISCLTFGR (SEQ ID NO: 21), CLTFGRETV (SEQ ID NO: 22) and LTFGRETVLEN (SEQ ID NO: 23), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 18-23.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising at least three, at least four, at least five or at least six hepatitis B virus (HBV) epitopes selected from YVNTNMGLK (SEQ ID NO: 18), GLKILQLL (SEQ ID NO: 19), LWFHISCLTF (SEQ ID NO: 20), HISCLTFGR (SEQ ID NO: 21), CLTFGRETV (SEQ ID NO: 22) and LTFGRETVLEN (SEQ ID NO: 23), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 18- 23.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three, at least four, at least five or at least six hepatitis B virus (HBV) epitopes selected from YVNTNMGLK (SEQ ID NO: 18), GLKILQLL (SEQ ID NO: 19), LWFHISCLTF (SEQ ID NO: 20), HISCLTFGR (SEQ ID NO: 21), CLTFGRETV (SEQ ID NO: 22) and LTFGRETVLEN (SEQ ID NO: 23), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 18-23, provided that the immunogenic peptide or immunogenic peptide group comprises at least one epitope restricted by human leukocyte antigen (HLA) A*02:01 and/or HLA A*24:02.
  • HLA human le
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising three, four, five or six hepatitis B virus (HBV) epitopes selected from YVNTNMGLK (SEQ ID NO: 18), GLKILQLL (SEQ ID NO: 19), LWFHISCLTF (SEQ ID NO: 20), HISCLTFGR (SEQ ID NO: 21), CLTFGRETV (SEQ ID NO: 22) and LTFGRETVLEN (SEQ ID NO: 23), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 18-23, provided that the immunogenic peptide or immunogenic peptide group comprises at least one sequence selected from CLTFGRETV (SEQ ID NO: 22) and LTFGRETVLEN (SEQ ID NO: 23).
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence YVNTNMGLKILQLLWFHISCLTFGRETVLENL (Cluster 3; SEQ ID NO: 2), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 2.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three, at least four, at least five or at least six hepatitis B virus (HBV) epitopes selected from SAICSVVRR (SEQ ID NO: 24), FPHCLAFSYM (SEQ ID NO: 25), YMDDVVLGV (SEQ ID NO: 26), YMDDVVLGA (SEQ ID NO: 27), YIDDVVLGA (SEQ ID NO: 28) and YVDDVVLGA (SEQ ID NO: 29), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 24-29.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising at least three, at least four, at least five or at least six hepatitis B virus (HBV) epitopes selected from SAICSVVRR (SEQ ID NO: 24), FPHCLAFSYM (SEQ ID NO: 25), YMDDVVLGV (SEQ ID NO: 26), YMDDVVLGA (SEQ ID NO: 27), YIDDVVLGA (SEQ ID NO: 28) and YVDDVVLGA (SEQ ID NO: 29), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 24- 29.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three, at least four, at least five or at least six hepatitis B virus (HBV) epitopes selected from SAICSVVRR (SEQ ID NO: 24), FPHCLAFSYM (SEQ ID NO: 25), YMDDVVLGV (SEQ ID NO: 26), YMDDVVLGA (SEQ ID NO: 27), YIDDVVLGA (SEQ ID NO: 28) and YVDDVVLGA (SEQ ID NO: 29), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 24-29, provided that the immunogenic peptide or immunogenic peptide group comprises at least one epitope restricted by human leukocyte antigen (HLA) A*02:01 and/or HLA A*24:02.
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three, at least four, at least five or at least six hepatitis B virus (HBV) epitopes selected from SAICSVVRR (SEQ ID NO: 24), FPHCLAFSYM (SEQ ID NO: 25), YMDDVVLGV (SEQ ID NO: 26), YMDDVVLGA (SEQ ID NO: 27), YIDDVVLGA (SEQ ID NO: 28) and YVDDVVLGA (SEQ ID NO: 29), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 24-29, provided that the immunogenic peptide or immunogenic peptide group comprises at least one sequence selected from YMDDVVLGV (SEQ ID No: 26), YMDDVVLGA (SEQ ID NO: 26
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence SAICSVVRRAFPHCLAFSYMDDVVLGV (Cluster 25; SEQ ID NO: 3), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 3.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three hepatitis B virus (HBV) epitopes selected from QAFTFSPTYK (SEQ ID NO: 30), FLKQQYMNL (SEQ ID NO: 31) and FLSKQYMDL (SEQ ID NO: 32), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 30-32.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising at least three hepatitis B virus (HBV) epitopes selected from QAFTFSPTYK (SEQ ID NO: 30), FLKQQYMNL (SEQ ID NO: 31) and FLSKQYMDL (SEQ ID NO: 32), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 30-32.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence QAFTFSPTYKAFLKQQYMNL (Cluster 28; SEQ ID NO: 4), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 4.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three hepatitis B virus (HBV) epitopes selected from FLLTRILTI (SEQ ID NO: 33), SWWTSLNFL (SEQ ID NO: 34) and FLGGPPVCL (SEQ ID NO: 35), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 33-35.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising at least three hepatitis B virus (HBV) epitopes selected from FLLTRILTI (SEQ ID NO: 33), SWWTSLNFL (SEQ ID NO: 34) and FLGGPPVCL (SEQ ID NO: 35), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 33-35.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence FFLLTRILTIPQSLDSWWTSLNFLGGAPTCP (Cluster 8; SEQ ID NO: 5), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 5.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three or at least four hepatitis B virus (HBV) epitopes selected from FLGPLLVLQA (SEQ ID NO: 36), VLQAGFFLL (SEQ ID NO: 37), FLLTRILTI (SEQ ID NO: 33) and SWWTSLNFL (SEQ ID NO: 34), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 36, 37, 33 and 34.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising at least three or at least four hepatitis B virus (HBV) epitopes selected from FLGPLLVLQA (SEQ ID NO: 36), VLQAGFFLL (SEQ ID NO: 37), FLLTRILTI (SEQ ID NO: 33) and SWWTSLNFL (SEQ ID NO: 34), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 36, 37, 33 and 34.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three or at least four hepatitis B virus (HBV) epitopes selected from FLGPLLVLQA (SEQ ID NO: 36), VLQAGFFLL (SEQ ID NO: 37), FLLTRILTI (SEQ ID NO: 33) and SWWTSLNFL (SEQ ID NO: 34), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 36, 37, 33 and 34, provided that the immunogenic peptide or immunogenic peptide group comprises at least one epitope restricted by human leukocyte antigen (HLA) A*02:01 and/or HLA A*24:02.
  • HLA human leukocyte antigen
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three or at least four hepatitis B virus (HBV) epitopes selected from FLGPLLVLQA (SEQ ID NO: 36), VLQAGFFLL (SEQ ID NO: 37), FLLTRILTI (SEQ ID NO: 33) and SWWTSLNFL (SEQ ID NO: 34), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 36, 37, 33 and 34, provided that the immunogenic peptide or immunogenic peptide group comprises at least one sequence selected from FLGPLLVLQA (SEQ ID No: 36), VLQAGFFLL (SEQ ID NO: 37) and FLLTRILTI (SEQ ID NO: 33).
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence FLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFL (Cluster 8B; SEQ ID NO: 6), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 6.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three, at least four, at least five or at least six hepatitis B virus (HBV) epitopes selected from RFSWLSLLVPF (SEQ ID NO: 38), SWLSLLVPF (SEQ ID NO: 39), WLSLLVPFV (SEQ ID NO: 40), VGLSPTVWL (SEQ ID NO: 41), GLSPTVWLS (SEQ ID NO: 42) and GLSPTVWLSV (SEQ ID NO: 43), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 38-43.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising at least three, at least four, at least five or at least six hepatitis B virus (HBV) epitopes selected from RFSWLSLLVPF (SEQ ID NO: 38), SWLSLLVPF (SEQ ID NO: 39), WLSLLVPFV (SEQ ID NO: 40), VGLSPTVWL (SEQ ID NO: 41), GLSPTVWLS (SEQ ID NO: 42) and GLSPTVWLSV (SEQ ID NO: 43), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 38-43.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three, at least four, at least five or at least six hepatitis B virus (HBV) epitopes selected from RFSWLSLLVPF (SEQ ID NO: 38), SWLSLLVPF (SEQ ID NO: 39), WLSLLVPFV (SEQ ID NO: 40), VGLSPTVWL (SEQ ID NO: 41), GLSPTVWLS (SEQ ID NO: 42) and GLSPTVWLSV (SEQ ID NO: 43), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 38-43, provided that the immunogenic peptide or immunogenic peptide group comprises at least one epitope restricted by human leukocyte antigen (HLA) A*02:01 and/or HLA A*24:02.
  • HLA human leukocyte
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three, at least four, at least five or at least six hepatitis B virus (HBV) epitopes selected from RFSWLSLLVPF (SEQ ID NO: 38), SWLSLLVPF (SEQ ID NO: 39), WLSLLVPFV (SEQ ID NO: 40), VGLSPTVWL (SEQ ID NO: 41), GLSPTVWLS (SEQ ID NO: 42) and GLSPTVWLSV (SEQ ID NO: 43), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 38-43, provided that the immunogenic peptide or immunogenic peptide group comprises at least one sequence selected from WLSLLVPFV (SEQ ID No: 40), GLSPTVWLS (SEQ ID NO: 42) and GLSPTVTV
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence SVRFSWLSLLVPFVQWFVGLSPTVWLSV (Cluster 13; SEQ ID NO: 7), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 7.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three hepatitis B virus (HBV) epitopes selected from VCWGELMNL (SEQ ID NO: 44), ELMTLATWV (SEQ ID NO: 45) and LATWVGSNL (SEQ ID NO: 46), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 44-46.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising at least three hepatitis B virus (HBV) epitopes selected from (SEQ ID NO: 44), ELMTLATWV (SEQ ID NO: 45) and LATWVGSNL (SEQ ID NO: 46), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 44-46.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence VCWGELMTLATWVGSNLEDPA (Cluster 2; SEQ ID NO: 8), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 8.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three hepatitis B virus (HBV) epitopes selected from STLPETTVV (SEQ ID NO: 47), STLPETTVVRR (SEQ ID NO: 48) and PSPRRRRS (SEQ ID NO: 49), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 47-49.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising at least three hepatitis B virus (HBV) epitopes selected from STLPETTVV (SEQ ID NO: 47), STLPETTVVRR (SEQ ID NO: 48) and PSPRRRRS (SEQ ID NO: 49), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 47-49.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence LSTLPETTVVRRRGRSPRRRTPSPRRRRSQ (Cluster 5; SEQ ID NO: 9), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 9.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three, at least four or at least five hepatitis B virus (HBV) epitopes selected from FLPSDFFPSV (SEQ ID NO: 12), LPSDFFPSV (SEQ ID NO: 13), EYLVSFGVW (SEQ ID NO: 50), STLPETTVV (SEQ ID NO: 47) and STLPETTVVRR (SEQ ID NO: 48), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 12, 13, 50, 47 and 48.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising at least three, at least four or at least five hepatitis B virus (HBV) epitopes selected from FLPSDFFPSV (SEQ ID NO: 12), LPSDFFPSV (SEQ ID NO: 13), EYLVSFGVW (SEQ ID NO: 50), STLPETTVV (SEQ ID NO: 47) and STLPETTVVRR (SEQ ID NO: 48), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 12, 13, 50, 47 and 48.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three, at least four or at least five hepatitis B virus (HBV) epitopes selected from FLPSDFFPSV (SEQ ID NO: 12), LPSDFFPSV (SEQ ID NO: 13), EYLVSFGVW (SEQ ID NO: 50), STLPETTVV (SEQ ID NO: 47) and STLPETTVVRR (SEQ ID NO: 48), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 12, 13, 50, 47 and 48, provided that the immunogenic peptide or immunogenic peptide group comprises at least one epitope restricted by human leukocyte antigen (HLA) A*02:01 and/or HLA A*24:02.
  • HLA human leukocyte antigen
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three, at least four or at least five hepatitis B virus (HBV) epitopes selected from FLPSDFFPSV (SEQ ID NO: 12), LPSDFFPSV (SEQ ID NO: 13), EYLVSFGVW (SEQ ID NO: 50), STLPETTVV (SEQ ID NO: 47) and STLPETTVVRR (SEQ ID NO: 48), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 12, 13, 50, 47 and 48, provided that the immunogenic peptide or immunogenic peptide group comprises at least one sequence selected from FLPSDFFPSV (SEQ ID NO: 12) and EYLVSFGVW (SEQ ID NO: 48).
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope sequence FLPSDFFPSVRLSTLPETTVVRRRLEYLVSFGVW (Bead 17; SEQ ID NO: 11), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 11.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three hepatitis B virus (HBV) epitopes selected from RLCCQLDPA (SEQ ID NO: 95), VLCLRPVGA (SEQ ID NO: 96) and RGRPVSGPF (SEQ ID NO: 97), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 95-97.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising at least three hepatitis B virus (HBV) epitopes selected from RLCCQLDPA (SEQ ID NO: 95), VLCLRPVGA (SEQ ID NO: 96) and RGRPVSGPF (SEQ ID NO: 97), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 111-113.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence RLCCQLDPARDVLCLRPVGAESRGRPVSGPF (Cluster 32; SEQ ID NO: 110), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 110.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least two hepatitis B virus (HBV) epitopes selected from FLLTRILTI (SEQ ID NO: 33) and SWWTSLNFL (SEQ ID NO: 34), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 33-34.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising at least two hepatitis B virus (HBV) epitopes selected from FLLTRILTI (SEQ ID NO: 33) and SWWTSLNFL (SEQ ID NO: 34), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 33-34.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence FFLLTRILTIPQSLDSWWTSLNFL (Cluster 8b REV 1; SEQ ID NO: 199), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 199.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least two or at least three hepatitis B virus (HBV) epitopes selected from VLQAGFFLL (SEQ ID NO: 37), FLLTRILTI (SEQ ID NO: 33) and SWWTSLNFL (SEQ ID NO: 34), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 37, 33 and 34.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising at least two or at least three hepatitis B virus (HBV) epitopes selected from VLQAGFFLL (SEQ ID NO: 37), FLLTRILTI (SEQ ID NO: 33) and SWWTSLNFL (SEQ ID NO: 34), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 37, 33 and 34.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence VLQAGFFLLTRILTIPQSLDSWWTSLNFL (Cluster 8b REV 2; SEQ ID NO: 200), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 200.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least two or at least three hepatitis B virus (HBV) epitopes selected from SWLSLLVPF (SEQ ID NO: 39), WLSLLVPFV (SEQ ID NO: 40) and VGLSPTVWL (SEQ ID NO: 41), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 39-41.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising at least two or at least three hepatitis B virus (HBV) epitopes selected from SWLSLLVPF (SEQ ID NO: 39), WLSLLVPFV (SEQ ID NO: 40) and VGLSPTVWL (SEQ ID NO: 41), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 39-41.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence SWLSLLVPFVQWFVGLSPTVWL (Cluster 13 REV 1; SEQ ID NO: 201), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 201.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three or at least four hepatitis B virus (HBV) epitopes selected from FLLTRILTI (SEQ ID NO: 33), SMYPSCCCTK (SEQ ID NO: 58), SWLSLLVPF (SEQ ID NO: 39) and WLSLLVPFV (SEQ ID NO: 40), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 33, 58, 39 and 40.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising at least three or at least four hepatitis B virus (HBV) epitopes selected from FLLTRILTI (SEQ ID NO: 33), SMYPSCCCTK (SEQ ID NO: 58), SWLSLLVPF (SEQ ID NO: 39) and WLSLLVPFV (SEQ ID NO: 40), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 33, 58, 39 and 40.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three or at least four hepatitis B virus (HBV) epitopes selected from FLLTRILTI (SEQ ID NO: 33), SMYPSCCCTK (SEQ ID NO: 58), SWLSLLVPF (SEQ ID NO: 39) and WLSLLVPFV (SEQ ID NO: 40), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 33, 58, 39 and 40, provided that the immunogenic peptide or immunogenic peptide group comprises at least one epitope restricted by human leukocyte antigen (HLA) A*02:01 and/or HLA A*24:02.
  • HLA human leukocyte antigen
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three or at least four hepatitis B virus (HBV) epitopes selected from FLLTRILTI (SEQ ID NO: 33), SMYPSCCCTK (SEQ ID NO: 58), SWLSLLVPF (SEQ ID NO: 39) and WLSLLVPFV (SEQ ID NO: 40), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 33, 58, 39 and 40, provided that the immunogenic peptide or immunogenic peptide group comprises at least one sequence selected from FLLTRILTI (SEQ ID NO: 33), SWLSLLVPF (SEQ ID NO: 39) and WLSLLVPFV (SEQ ID NO: 40).
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence FLLTRILTISMYPSCCCTKSWLSLLVPFV (E new bead 1; SEQ ID NO: 202), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 202.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least two or at least three hepatitis B virus (HBV) epitopes selected from FLGPLLVLQA (SEQ ID NO: 36), SWWTSLNFL (SEQ ID NO: 34) and GLSPTVWLSV (SEQ ID NO: 43), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 36, 34 and 43.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising at least two or at least three hepatitis B virus (HBV) epitopes selected from FLGPLLVLQA (SEQ ID NO: 36), SWWTSLNFL (SEQ ID NO: 34) and GLSPTVWLSV (SEQ ID NO: 43), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 36, 34 and 43.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence FLGPLLVLQASWWTSLNFLGLSPTVWLSV (E new bead 2; SEQ ID NO: 203), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 203.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least two or at least three hepatitis B virus (HBV) epitopes selected from LVLLDYQGML (SEQ ID NO: 205), SMYPSCCCTK (SEQ ID NO: 58) and SWLSLLVPF (SEQ ID NO: 39), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 205, 58 and 39.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising at least two or at least three hepatitis B virus (HBV) epitopes selected from LVLLDYQGML (SEQ ID NO: 205), SMYPSCCCTK (SEQ ID NO: 58) and SWLSLLVPF (SEQ ID NO: 39), or a variant sequence which shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 205, 58 and 39.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence LVLLDYQGMLSMYPSCCCTKSWLSLLVPF (E new bead 3; SEQ ID NO: 204), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 204.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence TVSTKLCKI (Cluster 31; SEQ ID NO: 143), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 143.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence KVGNFTGLYSSTVPV (Cluster 17; SEQ ID NO: 144), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 144.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence IPIPSSWAFARFLWEWASVR (Cluster 12; SEQ ID NO: 145), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 145.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence SAIWMMWYWGPSLYNILSPFLPLL (Cluster 14; SEQ ID NO: 146), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 146.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence EYLVSFGVWIR (Cluster 4; SEQ ID NO: 147), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 147.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence GYRWMCLRRFII (Cluster 9; SEQ ID NO: 148), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 148.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence YPALMPLYACI (Cluster 27; SEQ ID NO: 149), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 149.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence KYTSFPWLLGCAANWILRGTSFVYVPSAL (Cluster 29; SEQ ID NO: 150), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 150.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence QYVGPLTVN (Cluster 18; SEQ ID NO: 151), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 151.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence GLSRYVARL (Cluster 23; SEQ ID NO: 152), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 152.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence FLLSLGIHL (Cluster 26; SEQ ID NO: 153), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 153.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence VLHKRTLGLSAMSTTDLEAYFKDCLFKDWEEL (Cluster 34; SEQ ID NO: 154), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 154.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence LLLCLIFLLVLLDYQGMLPV (Cluster 10; SEQ ID NO: 155), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 155.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence SLYADSPSV (Cluster 30; SEQ ID NO: 156), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 156.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence FLGPLLVLQAGFFLL (Cluster 7; SEQ ID NO: 157), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 157.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence KLHLYSHPIIL (Cluster 24; SEQ ID NO: 158), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 158.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence HLSLRGLFV (Cluster 33; SEQ ID NO: 159), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 159.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence KLVCSPAPC (Cluster 35; SEQ ID NO: 160), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 160.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence LLGWSPQAQGILTTLPAAPPPA (Cluster 6; SEQ ID NO: 161), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 161.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence AILSKTGDPV (Cluster 15; SEQ ID NO: 162), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 162.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence GGSYFMISR (Cluster 16; SEQ ID NO: 163), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 163.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence NLSWLSLDV (Cluster 22; SEQ ID NO: 164), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 164.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence HTLWKAGILYKRETTRSASFCGSPYSW (Cluster 19; SEQ ID NO: 165), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 165.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence TPARVTGGVF (Cluster 20; SEQ ID NO: 166), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 166.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence LVVDFSQFSR (Cluster 21; SEQ ID NO: 167), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 167.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence SMYPSCCCTK (Cluster 11; SEQ ID NO: 168), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 168.
  • HBV hepatitis B virus
  • the present invention provides a composition comprising an immunogenic peptide or an immunogenic peptide group as described herein.
  • the present invention provides a composition comprising an immunogenic peptide or an immunogenic peptide group as described herein, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides a composition comprising at least one immunogenic peptide defined by LSFLPSDFFPSVRDLLDTASALY (Cluster 1; SEQ ID NO: 1), YVNTNMGLKILQLLWFHISCLTFGRETVLENL (Cluster 3; SEQ ID NO: 2), SAICSVVRRAFPHCLAFSYMDDVVLGV (Cluster 25; SEQ ID NO: 3), QAFTFSPTYKAFLKQQYMNL (Cluster 28; SEQ ID NO: 4), FFLLTRILTIPQSLDSWWTSLNFLGGAPTCP (Cluster 8; SEQ ID NO:
  • VCWGELMTLATWVGSNLEDPA Cluster 2; SEQ ID NO: 8
  • FLPSDFFPSVRLSTLPETTVVRRRLEYLVSFGVW (Bead 17; SEQ ID NO: 11), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 1-11, and optionally a pharmacologically acceptable excipient or carrier, and includes all combinations thereof.
  • the present invention provides a vaccine composition comprising at least three, at least four and at least five immunogenic peptides defined by LSFLPSDFFPSVRDLLDTASALY (Cluster 1; SEQ ID 1),
  • YVNTNMGLKILQLLWFHISCLTFGRETVLENL (Cluster 3; SEQ ID NO: 2), SAICSVVRRAFPHCLAFSYMDDVVLGV (Cluster 25; SEQ ID NO: 3), QAFTFSPTYKAFLKQQYMNL (Cluster 28; SEQ ID NO: 4), FFLLTRILTIPQSLDSWWTSLNFLGGAPTCP (Cluster 8; SEQ ID NO: 5), FLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFL (Cluster 8B; SEQ ID NO: 6), SVRFSWLSLLVPFVQWFVGLSPTVWLSV (Cluster 13; SEQ ID NO: 7), VCWGELMTLATWVGSNLEDPA (Cluster 2; SEQ NO: 8), LSTLPETTVVRRRGRSPRRRTPSPRRRRSQ (Cluster 5; SEQ ID NO: 9), FLPSDFFPSVRDLLDTASALY (Cluster 1A; SEQ ID NO: 10) and
  • FLPSDFFPSVRLSTLPETTVVRRRLEYLVSFGVW (Bead 17; SEQ ID NO: 11), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID NOs: 1-11, and optionally a pharmacologically acceptable excipient or carrier, and includes all combinations thereof.
  • the present invention provides a vaccine composition
  • a vaccine composition comprising at least one immunogenic peptide defined by LSFLPSDFFPSVRDLLDTASALY (Cluster 1; SEQ ID NO: 1), YVNTNMGLKILQLLWFHISCLTFGRETVLENL (Cluster 3; SEQ ID NO: 2), SAICSVVRRAFPHCLAFSYMDDVVLGV (Cluster 25; SEQ ID NO: 3), QAFTFSPTYKAFLKQQYMNL (Cluster 28; SEQ ID NO: 4), FFLLTRILTIPQSLDSWWTSLNFLGGAPTCP (Cluster 8; SEQ ID NO: 5), FLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFL (Cluster 8B; ID NO: 6), SVRFSWLSLLVPFVQWFVGLSPTVWLSV (Cluster 13; SEQ NO: 7), VCWGELMTLATWVGSNLEDPA (Cluster 2; SEQ ID NO: 8), LSTLPETTVVRRR
  • FLPSDFFPSVRLSTLPETTVVRRRLEYLVSFGVW (Bead 17; SEQ ID NO: 11), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID NOs: 1-11, and optionally a pharmacologically acceptable excipient or carrier, and includes all combinations thereof.
  • the present invention provides a vaccine composition comprising at least three, at least four and at least five immunogenic peptides defined by LSFLPSDFFPSVRDLLDTASALY (Cluster 1; SEQ ID NO: 1), YVNTNMGLKILQLLWFHISCLTFGRETVLENL (Cluster 3; SEQ ID NO: 2),
  • SAICSVVRRAFPHCLAFSYMDDVVLGV Cluster 25; SEQ ID NO: 3), QAFTFSPTYKAFLKQQYMNL (Cluster 28; SEQ ID NO: 4), FFLLTRILTIPQSLDSWWTSLNFLGGAPTCP (Cluster 8; SEQ ID NO: 5), FLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFL (Cluster 8B; SEQ ID NO: 6), SVRFSWLSLLVPFVQWFVGLSPTVWLSV (Cluster 13; SEQ ID NO: 7), VCWGELMTLATWVGSNLEDPA (Cluster 2; SEQ ID NO: 8), LSTLPETTVVRRRGRSPRRRTPSPRRRRSQ (Cluster 5; SEQ ID NO: 9), FLPSDFFPSVRDLLDTASALY (Cluster 1A; SEQ ID NO: 10) and FLPSDFFPSVRLSTLPETTVVRRRLEYLVSFGVW (Bead 17; SEQ ID NO: 11), or a variant
  • the present invention provides vaccine composition
  • vaccine composition comprising an immunogenic peptide defined by LSFLPSDFFPSVRDLLDTASALY (Cluster 1; SEQ ID NO: 1), an immunogenic peptide defined by YVNTNMGLKILQLLWFHISCLTFGRETVLENL (Cluster 3; SEQ ID NO: 2) and an immunogenic peptide defined by SAICSVVRRAFPHCLAFSYMDDVVLGV (Cluster 25; SEQ ID NO: 3), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides a vaccine composition
  • a vaccine composition comprising an immunogenic peptide defined by LSFLPSDFFPSVRDLLDTASALY (Cluster 1; SEQ ID NO: 1), an immunogenic peptide defined by YVNTNMGLKILQLLWFHISCLTFGRETVLENL (Cluster 3; SEQ ID NO: 2) and an immunogenic peptide defined by QAFTFSPTYKAFLKQQYMNL (Cluster 28; SEQ ID NO: 4), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 4, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • an immunogenic peptide defined by QAFTFSPTYKAFLKQQYMNL Cluster 28; SEQ ID NO: 4
  • an immunogenic peptide defined by VCWGELMTLATWVGSNLEDPA Cluster 2; SEQ ID NO: 8
  • an immunogenic peptide defined by LSFLPSDFFPSVRDLLDTASALY Cluster 1; SEQ ID NO: 1
  • a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 4, SEQ ID NO: 8 and SEQ ID NO: 1, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • an immunogenic peptide defined by QAFTFSPTYKAFLKQQYMNL Cluster 28; SEQ ID NO: 4
  • an immunogenic peptide defined by VCWGELMTLATWVGSNLEDPA Cluster 2; SEQ ID NO: 8
  • an immunogenic peptide defined by FLPSDFFPSVRLSTLPETTVVRRRLEYLVSFGVW Bead 17; SEQ ID NO: 11
  • a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 4, SEQ ID NO: 8 and SEQ ID NO: 11, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • an immunogenic peptide defined by QAFTFSPTYKAFLKQQYMNL Cluster 28; SEQ ID NO: 4
  • an immunogenic peptide defined by VCWGELMTLATWVGSNLEDPA Cluster 2; SEQ ID NO: 8
  • an immunogenic peptide defined by SVRFSWLSLLVPFVQWFVGLSPTVWLSV Cluster 13; SEQ ID NO: 7
  • a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 4, SEQ ID NO: 8 and SEQ ID NO: 7, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • an immunogenic peptide defined by QAFTFSPTYKAFLKQQYMNL Cluster 28; SEQ ID NO: 4
  • an immunogenic peptide defined by LSFLPSDFFPSVRDLLDTASALY Cluster 1; SEQ ID NO: 1
  • an immunogenic peptide defined by FLPSDFFPSVRLSTLPETTVVRRRLEYLVSFGVW Bead 17; SEQ ID NO: 11
  • a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 4, SEQ ID NO: 1 and SEQ ID NO: 11, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • an immunogenic peptide defined by QAFTFSPTYKAFLKQQYMNL Cluster 28; SEQ ID NO: 4
  • an immunogenic peptide defined by LSFLPSDFFPSVRDLLDTASALY Cluster 1; SEQ ID NO: 1
  • an immunogenic peptide defined by SVRFSWLSLLVPFVQWFVGLSPTVWLSV Cluster 13; SEQ ID NO: 7
  • a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 4, SEQ ID NO: 1 and SEQ ID NO: 7, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • the present invention provides vaccine composition
  • an immunogenic peptide defined by VCWGELMTLATWVGSNLEDPA Cluster 2; SEQ ID NO: 8
  • an immunogenic peptide defined by LSFLPSDFFPSVRDLLDTASALY Cluster 1; SEQ ID NO: 1
  • an immunogenic peptide defined by FLPSDFFPSVRLSTLPETTVVRRRLEYLVSFGVW Bead 17; SEQ ID NO: 11
  • a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 8, SEQ ID NO: 1 and SEQ ID NO: 11, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • an immunogenic peptide defined by VCWGELMTLATWVGSNLEDPA Cluster 2; SEQ ID NO: 8
  • an immunogenic peptide defined by LSFLPSDFFPSVRDLLDTASALY Cluster 1; SEQ ID NO: 1
  • an immunogenic peptide defined by SVRFSWLSLLVPFVQWFVGLSPTVWLSV Cluster 13; SEQ ID NO: 7
  • a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 8, SEQ ID NO: 1 and SEQ ID NO: 7, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • an immunogenic peptide defined by VCWGELMTLATWVGSNLEDPA Cluster 2; SEQ ID NO: 8
  • an immunogenic peptide defined by FLPSDFFPSVRLSTLPETTVVRRRLEYLVSFGVW Bead 17; SEQ ID NO: 11
  • an immunogenic peptide defined by SVRFSWLSLLVPFVQWFVGLSPTVWLSV Cluster 13; SEQ ID NO: 7
  • a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 8, SEQ ID NO: 11 and SEQ ID NO: 7, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • an immunogenic peptide defined by LSFLPSDFFPSVRDLLDTASALY (Cluster 1; SEQ ID NO: 1), an immunogenic peptide defined by FLPSDFFPSVRLSTLPETTVVRRRLEYLVSFGVW (Bead 17; SEQ ID NO: 11) and an immunogenic peptide defined by SVRFSWLSLLVPFVQWFVGLSPTVWLSV (Cluster 13; SEQ ID NO: 7), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 1, SEQ ID NO: 11 and SEQ ID NO: 7, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • an immunogenic peptide defined by QAFTFSPTYKAFLKQQYMNL Cluster 28; SEQ ID NO: 4
  • an immunogenic peptide defined by VCWGELMTLATWVGSNLEDPA Cluster 2; SEQ ID NO: 8
  • an immunogenic peptide defined by LSFLPSDFFPSVRDLLDTASALY Cluster 1; SEQ ID NO: 1
  • an immunogenic peptide defined by FLPSDFFPSVRLSTLPETTVVRRRLEYLVSFGVW Bead 17; SEQ ID NO: 11
  • a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 4, SEQ ID NO: 8, SEQ ID NO: 1 and SEQ ID NO: 11, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • an immunogenic peptide defined by QAFTFSPTYKAFLKQQYMNL Cluster 28; SEQ ID NO: 4
  • an immunogenic peptide defined by VCWGELMTLATWVGSNLEDPA Cluster 2; SEQ ID NO: 8
  • an immunogenic peptide defined by LSFLPSDFFPSVRDLLDTASALY Cluster 1; SEQ ID NO: 1
  • an immunogenic peptide defined by SVRFSWLSLLVPFVQWFVGLSPTVWLSV Cluster 13; SEQ ID NO: 7
  • a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 4, SEQ ID NO: 8, SEQ ID NO: 1 and SEQ ID NO: 7, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • an immunogenic peptide defined by QAFTFSPTYKAFLKQQYMNL Cluster 28; SEQ ID NO: 4
  • an immunogenic peptide defined by VCWGELMTLATWVGSNLEDPA Cluster 2; SEQ ID NO: 8
  • an immunogenic peptide defined by FLPSDFFPSVRLSTLPETTVVRRRLEYLVSFGVW Bead 17; SEQ ID NO: 11
  • an immunogenic peptide defined by SVRFSWLSLLVPFVQWFVGLSPTVWLSV (Cluster 13; SEQ ID NO: 7)
  • a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 4, SEQ ID NO: 8, SEQ ID NO: 11 and SEQ ID NO: 7, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • an immunogenic peptide defined by QAFTFSPTYKAFLKQQYMNL Cluster 28; SEQ ID NO: 4
  • an immunogenic peptide defined by LSFLPSDFFPSVRDLLDTASALY Cluster 1; SEQ ID NO: 1
  • an immunogenic peptide defined by FLPSDFFPSVRLSTLPETTVVRRRLEYLVSFGVW Bead 17; SEQ ID NO: 11
  • an immunogenic peptide defined by SVRFSWLSLLVPFVQWFVGLSPTVWLSV (Cluster 13; SEQ ID NO: 7)
  • a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 4, SEQ ID NO: 1, SEQ ID NO: 11 and SEQ ID NO: 7, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • an immunogenic peptide defined by VCWGELMTLATWVGSNLEDPA Cluster 2; SEQ ID NO: 8
  • an immunogenic peptide defined by LSFLPSDFFPSVRDLLDTASALY Cluster 1; SEQ ID NO: 1
  • an immunogenic peptide defined by FLPSDFFPSVRLSTLPETTVVRRRLEYLVSFGVW Bead 17; SEQ ID NO: 11
  • an immunogenic peptide defined by SVRFSWLSLLVPFVQWFVGLSPTVWLSV (Cluster 13; SEQ ID NO: 7)
  • a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 8, SEQ ID NO: 1, SEQ ID NO: 11 and SEQ ID NO: 7, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • an immunogenic peptide defined by QAFTFSPTYKAFLKQQYMNL Cluster 28; SEQ ID NO: 4
  • an immunogenic peptide defined by VCWGELMTLATWVGSNLEDPA Cluster 2; SEQ ID NO: 8
  • an immunogenic peptide defined by LSFLPSDFFPSVRDLLDTASALY Cluster 1; SEQ ID NO: 1
  • an immunogenic peptide defined by FLPSDFFPSVRLSTLPETTVVRRRLEYLVSFGVW Bead 17; SEQ ID NO: 11
  • an immunogenic peptide defined by SVRFSWLSLLVPFVQWFVGLSPTVWLSV Cluster 13; SEQ ID NO: 7
  • the present invention provides vaccine composition
  • vaccine composition comprising an immunogenic peptide defined by QAFTFSPTYKAFLKQQYMNL (Cluster 28; SEQ ID NO: 4), an immunogenic peptide defined by VCWGELMTLATWVGSNLEDPA (Cluster 2; SEQ ID NO: 8) and an immunogenic peptide defined by LSFLPSDFFPSVRDLLDTASALY (Cluster 1; SEQ ID NO: 1), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 4, SEQ ID NO: 8 and SEQ ID NO: 1, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • vaccine composition comprising an immunogenic peptide defined by QAFTFSPTYKAFLKQQYMNL (Cluster 28; SEQ ID NO: 4), an immunogenic peptide defined by VCWGELMTLATWVGSNLEDPA (Cluster 2; SEQ ID NO: 8) and an immunogenic peptide defined by FLPSDFFPSVRLSTLPETTVVRRRLEYLVSFGVW (Bead 17; SEQ ID NO: 11), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 4, SEQ ID NO: 8 and SEQ ID NO: 11, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • vaccine composition comprising an immunogenic peptide defined by QAFTFSPTYKAFLKQQYMNL (Cluster 28; SEQ ID NO: 4), an immunogenic peptide defined by VCWGELMTLATWVGSNLEDPA (Cluster 2; SEQ ID NO: 8) and an immunogenic peptide defined by FLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFL (Cluster 8B; SEQ ID NO: 6), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 4, SEQ ID NO: 8 and SEQ ID NO: 6, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • vaccine composition comprising an immunogenic peptide defined by QAFTFSPTYKAFLKQQYMNL (Cluster 28; SEQ ID NO: 4), an immunogenic peptide defined by LSFLPSDFFPSVRDLLDTASALY (Cluster 1; SEQ ID NO: 1) and an immunogenic peptide defined by FLPSDFFPSVRLSTLPETTVVRRRLEYLVSFGVW (Bead 17; SEQ ID NO: 11), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 4, SEQ ID NO: 1 and SEQ ID NO: 11, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • vaccine composition comprising an immunogenic peptide defined by QAFTFSPTYKAFLKQQYMNL (Cluster 28; SEQ ID NO: 4), an immunogenic peptide defined by LSFLPSDFFPSVRDLLDTASALY (Cluster 1; SEQ ID NO: 1) and an immunogenic peptide defined by FLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFL (Cluster 8B; SEQ ID NO: 6), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 4, SEQ ID NO: 1 and SEQ ID NO: 6, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • vaccine composition comprising an immunogenic peptide defined by QAFTFSPTYKAFLKQQYMNL (Cluster 28; SEQ ID NO: 4), an immunogenic peptide defined by FLPSDFFPSVRLSTLPETTVVRRRLEYLVSFGVW (Bead 17; SEQ ID NO: 11) and an immunogenic peptide defined by FLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFL (Cluster 8B; SEQ ID NO: 6), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 4, SEQ ID NO: 11 and SEQ ID NO: 6, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • vaccine composition comprising an immunogenic peptide defined by VCWGELMTLATWVGSNLEDPA (Cluster 2; SEQ ID NO: 8), an immunogenic peptide defined by LSFLPSDFFPSVRDLLDTASALY (Cluster 1; SEQ ID NO: 1) and an immunogenic peptide defined by FLPSDFFPSVRLSTLPETTVVRRRLEYLVSFGVW (Bead 17; SEQ ID NO: 11), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 8, SEQ ID NO: 1 and SEQ ID NO: 11, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • vaccine composition comprising an immunogenic peptide defined by VCWGELMTLATWVGSNLEDPA (Cluster 2; SEQ ID NO: 8), an immunogenic peptide defined by LSFLPSDFFPSVRDLLDTASALY (Cluster 1; SEQ ID NO: 1) and an immunogenic peptide defined by FLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFL (Cluster 8B; SEQ ID NO: 6), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 8, SEQ ID NO: 1 and SEQ ID NO: 6, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • vaccine composition comprising an immunogenic peptide defined by VCWGELMTLATWVGSNLEDPA (Cluster 2; SEQ ID NO: 8), an immunogenic peptide defined by FLPSDFFPSVRLSTLPETTVVRRRLEYLVSFGVW (Bead 17; SEQ ID NO: 11) and an immunogenic peptide defined by FLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFL (Cluster 8B; SEQ ID NO: 6), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 8, SEQ ID NO: 11 and SEQ ID NO: 6, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • vaccine composition comprising an immunogenic peptide defined by LSFLPSDFFPSVRDLLDTASALY (Cluster 1; SEQ ID NO: 1), an immunogenic peptide defined by FLPSDFFPSVRLSTLPETTVVRRRLEYLVSFGVW (Bead 17; SEQ ID NO: 11) and an immunogenic peptide defined by FLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFL (Cluster 8B; SEQ ID NO: 6), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 1, SEQ ID NO: 11 and SEQ ID NO: 6, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • vaccine composition comprising an immunogenic peptide defined by QAFTFSPTYKAFLKQQYMNL (Cluster 28; SEQ ID NO: 4), an immunogenic peptide defined by VCWGELMTLATWVGSNLEDPA (Cluster 2; SEQ ID NO: 8), an immunogenic peptide defined by LSFLPSDFFPSVRDLLDTASALY (Cluster 1; SEQ ID NO: 1) and an immunogenic peptide defined by FLPSDFFPSVRLSTLPETTVVRRRLEYLVSFGVW (Bead 17; SEQ ID NO: 11), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 4, SEQ ID NO: 8 and SEQ ID NO: 1 and SEQ ID NO: 11, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • vaccine composition comprising an immunogenic peptide defined by QAFTFSPTYKAFLKQQYMNL (Cluster 28; SEQ ID NO: 4), an immunogenic peptide defined by VCWGELMTLATWVGSNLEDPA (Cluster 2; SEQ ID NO: 8), an immunogenic peptide defined by LSFLPSDFFPSVRDLLDTASALY (Cluster 1; SEQ ID NO: 1) and an immunogenic peptide defined by FLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFL (Cluster 8B; SEQ ID NO: 6), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 4, SEQ ID NO: 8 and SEQ ID NO: 1 and SEQ ID NO: 6, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • vaccine composition comprising an immunogenic peptide defined by QAFTFSPTYKAFLKQQYMNL (Cluster 28; SEQ ID NO: 4), an immunogenic peptide defined by VCWGELMTLATWVGSNLEDPA (Cluster 2; SEQ ID NO: 8), an immunogenic peptide defined by FLPSDFFPSVRLSTLPETTVVRRRLEYLVSFGVW (Bead 17; SEQ ID NO: 11) and an immunogenic peptide defined by FLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFL (Cluster 8B; SEQ ID NO: 6), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 4, SEQ ID NO: 8 and SEQ ID NO: 11 and SEQ ID NO: 6, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • an immunogenic peptide defined by QAFTFSPTYKAFLKQQYMNL Cluster 28; SEQ ID NO: 4
  • an immunogenic peptide defined by LSFLPSDFFPSVRDLLDTASALY Cluster 1; SEQ ID NO: 1
  • an immunogenic peptide defined by FLPSDFFPSVRLSTLPETTVVRRRLEYLVSFGVW Bead 17; SEQ ID NO: 11
  • an immunogenic peptide defined by FLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFL Cluster 8B; SEQ ID NO: 6
  • a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 4, SEQ ID NO: 1 and SEQ ID NO: 11 and SEQ ID NO: 6, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • vaccine composition comprising an immunogenic peptide defined by VCWGELMTLATWVGSNLEDPA (Cluster 2; SEQ ID NO: 8), an immunogenic peptide defined by LSFLPSDFFPSVRDLLDTASALY (Cluster 1; SEQ ID NO: 1), an immunogenic peptide defined by FLPSDFFPSVRLSTLPETTVVRRRLEYLVSFGVW (Bead 17; SEQ ID NO: 11) and an immunogenic peptide defined by FLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFL (Cluster 8B; SEQ ID NO: 6), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 8, SEQ ID NO: 1 and SEQ ID NO: 11 and SEQ ID NO: 6, and optionally a pharmacologically acceptable excipient or carrier.
  • the present invention provides vaccine composition
  • an immunogenic peptide defined by QAFTFSPTYKAFLKQQYMNL Cluster 28; SEQ ID NO: 4
  • an immunogenic peptide defined by VCWGELMTLATWVGSNLEDPA Cluster 2; SEQ ID NO: 8
  • an immunogenic peptide defined by LSFLPSDFFPSVRDLLDTASALY Cluster 1; SEQ ID NO: 1
  • an immunogenic peptide defined by FLPSDFFPSVRLSTLPETTVVRRRLEYLVSFGVW Bead 17; SEQ ID NO: 11
  • an immunogenic peptide defined by FLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFL Cluster 8B; SEQ ID NO: 6
  • a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 4, SEQ ID NO: 8 and SEQ ID NO:
  • the present invention provides a method for treating acute or chronic hepatitis B virus (HBV) infection in a human subject in need thereof, the method comprising administering to the human subject at least one composition or vaccine composition as described herein.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group as described herein for use in treating acute or chronic hepatitis B virus infection in a human subject requiring treatment.
  • the present invention provides a vaccine composition as described herein for use in treating acute or chronic hepatitis B virus infection in a human subject requiring treatment.
  • the present invention provides an immunogenic peptide, an immunogenic peptide group or a vaccine composition as described herein for the manufacture of a medicament for treating acute or chronic hepatitis B virus infection in a human subject requiring treatment.
  • the present invention provides the use of an immunogenic peptide, an immunogenic peptide group or a vaccine composition as described herein for treating acute or chronic hepatitis B virus infection in a human subject requiring treatment.
  • the present invention provides a method for generating a peptide- loaded antigen-presenting cell, the method comprising contacting an antigen-presenting cell with :
  • the present invention provides a method of autologous cell therapy in a patient infected with a hepatitis B virus, the method comprising : (i) purifying at least one antigen-presenting cell from a sample obtained from the patient;
  • Figure 1 lists the hepatitis B virus HLA class I restricted epitopes considered for inclusion in the immunogenic peptide compositions and immunogenic peptides described herein.
  • the first three columns show the epitope sequence, the corresponding viral antigen it is derived from, and starting position.
  • the next set of columns indicate the number of references in the published literature that reported the epitope, followed by the number of positive responses (over the total number of assays), compiled considering all of the various HLA restrictions associated with each epitope. These HLA restrictions are compiled in the next (“HLA restriction”) column.
  • Figure 2 lists the frequency of restricting alleles for the hepatitis B virus HLA class I restricted epitopes considered for inclusion in the immunogenic peptide/compositions described herein.
  • Figure 3 shows the population coverage (i.e.) number of epitope/HLA binding interactions recognised for the cluster constructs defined by SEQ ID Nos: 2-5 and 7-10 in the Caucasian population.
  • Figure 4 shows the population coverage (i.e.) number of epitope/HLA binding interactions recognised for the cluster constructs defined by SEQ ID Nos: 2-4 in the Caucasian population.
  • Figure 5 shows human blood PBMC assay data for the various constructs defined herein as Cluster 1 (SEQ ID NO: 1), Cluster 3 (SEQ ID NO: 2), Cluster 25 (SEQ ID NO: 3), Cluster 28 (SEQ ID NO: 4), Cluster 8B (SEQ ID NO: 6), Cluster 13 (SEQ ID NO: 7), Cluster 2 (SEQ ID NO: 8), Cluster 32 (SEQ ID NO: 110) and Bead 17 (SEQ ID NO: 11).
  • Cluster 1 SEQ ID NO: 1
  • Cluster 3 SEQ ID NO: 2
  • Cluster 25 SEQ ID NO: 3
  • Cluster 28 SEQ ID NO: 4
  • Cluster 8B SEQ ID NO: 6
  • Cluster 13 SEQ ID NO: 7
  • Cluster 2 SEQ ID NO: 8
  • Cluster 32 SEQ ID NO: 110
  • Bead 17 SEQ ID NO: 11
  • a or “an” refers to one or more than one of the entity specified; for example, "a receptor” or “a nucleic acid molecule” may refer to one or more receptor or nucleic acid molecule, or at least one receptor or nucleic acid molecule. As such, the terms “a” or “an”, “one or more” and “at least one” can be used interchangeably herein.
  • composition of matter, group of steps or group of compositions of matter shall be taken to encompass one and a plurality (i.e. one or more) of those steps, compositions of matter, groups of steps or group of compositions of matter.
  • antigen refers to a molecule capable of inducing an immune response in a host organism, typically a mammalian host organism, such as a human host.
  • epitope refers to a region or part of an antigen, such as a immunogenic peptide disclosed herein, that elicits an immune response when administered to a subject.
  • An epitope may be a T-cell epitope, i.e., an epitope that elicits, stimulates, induces, promotes, increases or enhances a T cell activity, function or response.
  • identity and “identical” and grammatical variations thereof, as used herein, mean that two or more referenced entities are the same (e.g., amino acid sequences). Thus, where two peptides are identical, they have the same amino acid sequence. The identity can be over a defined area, e.g. over at least 22, 23, 24, 25 or 26 contiguous amino acids of the parent peptide sequence or whichever alignment is the best fit with gaps permitted.
  • Identity can be determined by comparing each position in aligned sequences.
  • a degree of identity between amino acid sequences is a function of the number of identical or matching amino acids at positions shared by the sequences, i.e. over a specified region.
  • Optimal alignment of sequences for comparisons of identity may be conducted using a variety of algorithms, as are known in the art, including the Clustal Omega program available at the website location at www.ebi.ac.uk/Tools/mas/clustalo/, the local homology algorithm of Smith and Waterman, 1981, Adv. Appl. Math 2: 482, the homology alignment algorithm of Needleman and Wunsch, 1970, J. Mol. Biol. 48:443, the search for similarity method of Pearson and Lipman, 1988, Proc. Natl.
  • Sequence identity may also be determined using the BLAST algorithm, described in Altschul et al., 1990, J. Mol. Biol. 215:403-10 (using the published default settings). Software for performing BLAST analysis may be available through the National Center for Biotechnology Information (through the internet at the website located at www.ncbi.nlm.nih.gov). Such algorithms that calculate percent sequence identity (homology) generally account for sequence gaps and mismatches over the comparison region or area.
  • a BLAST e.g., BLAST 2.0 search algorithm
  • BLAST 2.0 search algorithm
  • exemplary search parameters as follows: Mismatch -2; gap open 5; gap extension 2.
  • a BLASTP algorithm is typically used in combination with a scoring matrix, such as PAM 100, PAM 250, BLOSUM 62 or BLOSUM 50.
  • FASTA e.g., FASTA2 and FASTA3
  • SSEARCH sequence comparison programs are also used to quantitate the extent of identity (Pearson et al., Proc. Natl. Acad. Sci.
  • peptide or “immunogenic peptide”, as used herein is intended to mean an individual (e.g. isolated) amino acid molecule having a sequence length of between about 10 and about 100 amino acid residues, preferably between about 10 and about 50 amino acid residues and more preferably between about 10 and about 35 amino acid residues, and comprising either discrete or overlapping HBV T-cell epitopes (or both).
  • a peptide as referred to herein may be a linear peptide, which does not comprise a secondary helix structure.
  • a peptide as used herein may be a parent peptide or a variant thereof.
  • immunogenic peptide group as used herein is intended to mean that selected HBV specific epitopes including (e.g.) T-cell epitopes are present on two or more immunogenic peptides as defined herein.
  • parent peptide denotes an individual peptide that represents a region of the hepatitis B virus antigen it originates from, which region contains at least one T-cell epitope that herein is reported to produce an in vitro T-cell response in a high fraction of the donor population and to have broad HLA class I coverage.
  • population coverage is intended to mean the percentage of individuals in any given population (e.g., "worldwide population") or a specific ethnicity population, such as “Caucasian population” or “Asian population”, which express at least one human leukocyte antigen (HLA) A or B allele known to restrict a selected set of T-cell epitopes. Further information concerning the calculations used herein is provided in the Detailed Description.
  • HLA human leukocyte antigen
  • the present invention is concerned with immunogenic peptides and vaccine compositions for the treatment and prevention of hepatitis B virus (HBV) infection.
  • HBV hepatitis B virus
  • the present invention is predicated on selection of specific subsets of human leukocyte antigen (HLA) class I restricted T-cell epitopes, and their combinatorial assembly in immunological peptide constructs and vaccine compositions which allow for maximal worldwide population coverage for the treatment of HBV infection.
  • HLA human leukocyte antigen
  • the present invention is also predicated on selection of specific subsets of human leukocyte antigen (HLA) class I restricted T-cell epitopes, and their combinatorial assembly in immunological peptide constructs and vaccine compositions which allow for targeted population coverage, for example by geography or ethnicity, for the treatment of HBV infection.
  • HLA human leukocyte antigen
  • T-cell epitopes In order to reach maximum population coverage in the human population it is necessary to incorporate several T-cell epitopes to account for high HLA diversity. This can be achieved by combining several epitopes in a single vaccine (i.e. combination of epitopes within the same peptide, separate peptides or any combination thereof) and/or by combining vaccine compositions.
  • HBV epitopes To select HBV epitopes for this purpose, an extensive review was conducted of HBV epitopes that have been defined in HBV patients, and a series of the best-evaluated epitopes were selected that, when appropriately combined, maximise population coverage based on known distribution of class I HLA alleles. The selection criteria governing epitope selection, as well as the rationale for their combinatorial assembly in immunogenic peptides/compositions, is described later in this section.
  • CD4 and CD8 T-cells recognise a complex between specific HLA molecules and pathogen derived (e.g. HBV) peptide epitopes.
  • Each individual expresses HLA molecules encoded by three different class I loci (A, B and C), and four class II loci (DRB1, DRB3/4/5, DP and DQ).
  • Each of these loci are very polymorphic, and when heterozygosity is taken into account each individual express about six different class I and eight different class II variants. It should also be understood that the frequency of the different alleles varies between different ethnicities, a factor that needs to be accounted for when projecting the coverage afforded by various epitope combinations.
  • Figure 1 lists the best evaluated HBV-derived HLA class I restricted T-cell epitopes (86 total) based on selection criteria which is detailed later.
  • Figure 2 lists the various alleles that have been experimentally documented to present each or any of the selected HBV-derived HLA class I restricted epitopes recognized by human T-cells.
  • Figure 2 further lists the frequency of each allele derived from data available in the Allele Frequency Net Database (AFND; www.3Helefrequencies.net).
  • AFND provides a central source for the storage of allele frequencies from different polymorphic areas in the Human Genome.
  • AFND is part of EUROSTAM (an FP7-HEALTH collaborative project) to facilitate transplantation of highly sensitized patients. Data in the AFND are derived from published literature and/or reviewed submissions.
  • HLA allele frequencies are presented as: (i) phenotype frequency, which is defined as the fraction of individuals expressing a particular allele, and (ii) gene frequency, which is defined as the fraction of all genes at a locus that are represented by the specific allelic variant. As a rule, the phenotype frequency is higher than the genotype frequency. To illustrate this point, assume that a given allele "HLA X" makes up for 10% of the genes.
  • Figure 2 also projects the combined gene and phenotype frequencies based on the alleles listed, meaning that based on the calculations above and the combined gene frequencies, it is expected that about 84.5% of individuals would express at least one of the HLA A alleles known to restrict the selected epitopes listed in Table 1, and that 37.2% of individuals would express at least one of the HLA B alleles known to restrict the selected epitopes also listed in Table 1. In total, 90.2% of individuals would express at least one of the HLA A or B alleles known to restrict the selected epitopes listed in Table 1.
  • the term "population coverage" defines the % of individuals that express at least one of the HLA A or B alleles known to restrict the selected epitopes listed in Figure 1.
  • the calculations described herein use an estimate based on the average frequency of the HLA alleles across 11 different geographic regions worldwide, as described above.
  • the term "worldwide population coverage” as used herein is defined as the average % of individuals across several broad geographic regions that express at least one of the HLA A or B alleles known to restrict a set of selected epitopes, for example, those epitopes listed in Figure 1.
  • the coverage projected in specific populations can be calculated, for example in Caucasian heritage or Asian populations.
  • Caucasian population coverage or “Caucasian coverage” defines the % of individuals in Caucasian populations that express at least one of the HLA A or B alleles known to restrict a set of selected epitopes, for example, those listed in Table 1.
  • the present invention further contemplates calculations for targeted population coverage which, according to the principles described above, defines the coverage afforded by a sub/set of selected epitopes, or the coverage afforded by at least one immunogenic peptide/composition incorporating a selected subset of epitopes which were inventively selected and assembled to maximize coverage while minimizing complexity.
  • Example 6 in respect of cluster 8B (SEQ ID NO: 6) and population coverage in respect of Southeast Asia.
  • the present invention further contemplates the coverage provided by each immunogenic peptide/composition described herein in terms of the percentage distribution of individuals in a population of a given ethnicity (or average worldwide) covered by a different number of epitopes/HLA combinations recognised.
  • depth of coverage refers to what fraction of individuals are expected to recognize one, two, or all three. This can be calculated if it is known that epitope xl binds (or is restricted to) HLA X, Y and/or Z; epitope x2 binds (or is restricted to) HLA X and K (but not Y or Z); and epitope x3 binds (or is restricted to) HLA W; and so on. Knowing the frequency of the HLA alleles in a particular population, the fraction of individuals which will bind (recognize) none of the peptides, how many will bind one or two, and how many will bind all three, can be calculated.
  • the IEDB (www.iedb.org) was queried for linear peptide sequences derived from HBV, with humans specified as the host organism, reported in the published literature as having the capacity to elicit an immune response restricted by human MHC class I molecules.
  • HLA transgenic mice were also included as an alternative source organism.
  • the query was limited to T-cell epitopes associated with positive responses with class I MHC restriction. This query returned 177 unique epitopes, associated with 901 different assays, reported in 118 publications/references.
  • various selection measures were applied to eliminate peptide sequences which did not meet pre-determined selection criteria. For example, excluded from consideration were epitope sequences that did not meet (e.g.) canonical peptide size for HLA class I ligands, assay validation thresholds such as type of assay and number of assays performed, T-cell source/stimulation used to detect the epitope response, as well as the elimination of variant epitope sequences determined to have a weaker immune response.
  • the in/exclusion selection criteria are outlined in more detail in Example 1.
  • epitope "clusters" encompassing stretches of approximately 32 contiguous amino acids.
  • Each epitope was aligned against a corresponding HBV reference protein sequence specific for different antigens and included the external core protein (P0C767, inclusive of the capsid protein), large envelope protein (Q76R62), protein P (Q69028) and protein X (Q69027). These represent the HBV reference sequence set utilised by the IEDB. The starting positions of all epitopes were indexed to the respective reference sequence, and the epitopes sorted accordingly.
  • clusters In total, 35 different clusters were identified. The clusters ranged in inclusiveness from just one epitope to as many six epitopes and ranged in length from eight residues up to 32 residues. Table 1 below lists the 35 clusters, sorted in descending order on the basis of estimated population coverage.
  • Clusters were found to be associated with population coverage >50%.
  • One of these clusters is from Protein X, which has not been associated with robust immunological responses, even in acutely infected patients.
  • Another cluster consists of a single epitope from Protein P which did not map to the Protein P reference sequence. Accordingly, these two clusters were removed from further consideration.
  • the remaining nine clusters includes all but one cluster with four or more unique HLA class I restrictions (i.e. "No. alleles"), and all clusters with more than 30 total responses.
  • Four of the clusters are derived from the capsid/external core protein, three of the clusters are derived from the envelope protein and two clusters are derived from protein P.
  • a dash indicates that the epitope did not align with the corresponding HBV reference sequence.
  • the highly stringent epitope selection protocol described above followed by the combinatorial assembly of selected epitopes into 20-35 contiguous amino acid peptides, has allowed the Applicants to conceive of novel vaccine candidates for the treatment of chronic hepatitis B virus infection.
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three, at least four, at least five or at least six hepatitis B virus (HBV) epitopes selected from the group of epitope sequences derived from Cluster 1 and as set forth in any one of SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16 and SEQ ID NO: 17.
  • HBV hepatitis B virus
  • variant epitope sequences may invoke an identical or similar immunogenic response to the parent epitopes from which they derive.
  • the epitope sequences defined by SEQ ID Nos: 12-17 include either nine or ten amino acid residues, whereas other epitope sequences include as few as eight amino acid residues (e.g. SEQ ID NOs: 19 and 49).
  • the variant epitope sequences disclosed herein may differ, for example, by one, two or as many as three residues.
  • the present invention therefore contemplates variant epitope sequences which share at least 75% sequence identity to the epitope sequences from which they derive.
  • 75% sequence identity to an HBV T-cell epitope sequence having 8 residues means the variant sequence may include up to two residue(s) as either a substitution, insertion or deletion.
  • Modification may arise by substitution of one or more residues at different, possibly selective, sites within any given epitope sequence or indeed immunogenic peptide per se.
  • Such substitutions may be of a conservative nature, for example, where one amino acid is replaced by an amino acid of similar structure and characteristics, such as where a hydrophobic amino acid is replaced by another hydrophobic amino acid. Even more conservative would be replacement of amino acids of the same or similar size and chemical nature, such as where leucine is replaced by isoleucine.
  • Conservative substitutions are herein defined as exchanges within one of the following five groups: Group 1— small aliphatic, nonpolar or slightly polar residues (Ala, Ser, Thr, Pro, Gly); Group 2— polar, negatively charged residues and their amides (Asp, Asn, Glu, Gin); Group 3— polar, positively charged residues (His, Arg, Lys); Group 4— large, aliphatic, nonpolar residues (Met, Leu, lie, Vai, Cys); and Group 4— large, aromatic residues (Phe, Tyr, Trp).
  • An acidic amino acid might also be substituted by a different acidic amino acid or a basic (i.e., alkaline) amino acid by a different basic amino acid.
  • substitutions may involve structures other than the common L-amino acids.
  • D-amino acids might be substituted for the L-amino acids commonly found in the antigenic peptides of the invention and yet still be encompassed by the disclosure herein.
  • amino acids possessing non-standard R groups i.e., R groups other than those found in the common 20 amino acids of natural proteins may also be used for substitution purposes to produce immunogens and immunogenic polypeptides according to the present invention.
  • substitutions at more than one position are found to result in a peptide with substantially equivalent or greater antigenic activity as defined below, then combinations of those substitutions will be tested to determine if the combined substitutions result in additive or syngeneic effects on the antigenicity of the peptide.
  • the present invention further provides an immunogenic peptide or an immunogenic peptide group comprising at least three, at least four, at least five or at least six hepatitis B virus (HBV) epitopes selected from a variant sequence which shares at least 75%, 80%, 85%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 12-17.
  • HBV hepatitis B virus
  • sequence identity includes, without limitation, the side-by-side comparison of two sequences to determine that the overall level of identity is at least 75%, or at least 80%, or at least 81%, or at least 82%, or at least 83%, or at least 84%, or at least 85%, or at least 86%, or at least 87%, or at least 88%, or at least 89%, or at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99% identical.
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three, at least four, at least five or at least six hepatitis B virus (HBV) epitopes selected from the group of epitope sequences derived from Cluster 3 and as set forth in any one of SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22 and SEQ ID NO: 23, or a variant sequence which shares at least 75%, 80%, 85%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 18-23.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three hepatitis B virus (HBV) epitopes derived from Cluster 28 and as set forth in any one of SEQ ID NO: 30, SEQ ID NO: 31 and SEQ ID NO: 32, or a variant sequence which shares at least 75%, 80%, 85%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 30-32.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three, at least four, at least five or at least six hepatitis B virus (HBV) epitopes selected from the group of epitope sequences derived from Cluster 25 and as set forth in any one of SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28 and SEQ ID NO: 29, or a variant sequence which shares at least 75%, 80%, 85%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 24-29.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three hepatitis B virus (HBV) epitopes derived from Cluster 8 and as set forth in any one of SEQ ID NO: 33, SEQ ID NO: 34 and SEQ ID NO: 35, or a variant sequence which shares at least 75%, 80%, 85%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 33-35.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three or at least four hepatitis B virus (HBV) epitopes selected from the group of epitope sequences derived from Cluster 8B and as set forth in any one of SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 33 and SEQ ID NO: 34, or a variant sequence which shares at least 75%, 80%, 85%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 36, 37, 33 and 34.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three, at least four, at least five or at least six hepatitis B virus (HBV) epitopes selected from the group of epitope sequences derived from Cluster 13 and as set forth in any one of SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42 and SEQ ID NO: 43, a variant sequence which shares at least 75%, 80%, 85%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 38-43.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three hepatitis B virus (HBV) epitopes derived from Cluster 2 and as set forth in any one of SEQ ID NO: 44, SEQ ID NO: 45 and SEQ ID NO: 46, or a variant sequence which shares at least 75%, 80%, 85%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 44-46.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three hepatitis B virus (HBV) epitopes derived from Cluster 5 and as set forth in any one of SEQ ID NO: 47, SEQ ID NO: 48 and SEQ ID NO: 49, or a variant sequence which shares at least 75%, 80%, 85%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 47-49.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three, at least four or at least five hepatitis B virus (HBV) epitopes derived from Bead 17 and as set forth in any one of SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 50, SEQ ID NO: 47 and SEQ ID NO: 48, or a variant sequence which shares at least 75%, 80%, 85%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 12, 13, 50, 47 and 48.
  • HBV hepatitis B virus
  • immunogenic peptides or immunogenic peptide group(s) described herein may further comprise at least one CD4 + T-helper cell epitope.
  • the CD4 + T-helper cell epitope is selected from LSTLPETTVVRRRGRS (SEQ ID NO: 98), LSFLPSDFFPSVRDLLDTA (SEQ ID NO: 99), CWGELMTLATWVGVNLEDPA (SEQ ID NO: 100), LLWFHISCLTFGRETVLEYL (SEQ ID NO: 101), LSTLPETTVVRRRGRSPRRRT (SEQ ID NO: 102), TVVRRRGRSPRRRTP (SEQ ID NO: 103), SPRRRTPSPRRRRSQ (SEQ ID NO: 104), FFLLTRILTIPQSLD (SEQ ID NO: 105), TRILTIPQSLDSWWTSLNF (SEQ ID NO: 106), SVRFSWLSLLVPFVQWF (SEQ ID NO: 107), SWLSLLVPFVQWFVGL (SEQ ID NO: 108) and SLLVPFVQWFVGLSPTVWLSV (SEQ ID NO: 109
  • the immunogenic peptides and immunogenic peptide groups comprise between about 20 and about 35 amino acid residues.
  • the term "between about 20 and about 35 amino acid residues” includes 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34 and 35 amino acids in length.
  • the term “about” is intended to encompass peptides that have, without limitation, 17, 18 or 19 amino acid residues or 36, 37, or 38 amino acid residues. The optimal length, and therefore precise designation of its size, will depend on the epitopes included within the peptide clusters and could be performed by routine experiment, including various assays and/or analyses described herein.
  • a peptide or a variant peptide thereof may be chemically modified or derivatised.
  • a derivative of a peptide refers to a modified form of a peptide disclosed herein, including a modified form of a variant thereof.
  • a derivative is formed by reacting a functional side group of an amino acid (e.g. amino, sulfhydryl or carboxy-group) with another molecule to form a covalent or non-covalent attachment of any type of molecule (naturally occurring or designed), such as a sugar moiety.
  • an amino acid e.g. amino, sulfhydryl or carboxy-group
  • Specific examples of derivatives of a peptide include glycosylation, acylation (e.g.
  • acetylation phosphorylation
  • amidation formylation
  • ubiquitination phosphorylation
  • derivatization by protecting/blocking groups and any of numerous chemical modifications.
  • Additional specific non-limiting examples are tagged peptides, fusion peptides, chimeric peptides including peptides having one or more non-amino acyl groups (q.v., sugar, lipid, etc.) covalently linked to the peptide.
  • a derivative comprises one or more modifications, for example selected from any of: (a) N-terminal acylation (e.g. acetylation or formylation); (b) C-terminal amidation (e.g.
  • reaction with ammonia or an amine (c) one or more hydrogens on the side chain amines of arginine and/or lysine replaced with a methylene group; (d) glycosylation and/or (e) phosphorylation.
  • the immunogenic peptides disclosed herein may also be linked directly to, or through, a spacer or linker to a vaccine adjuvant such as alpha galactose ceramide (o-GC), Toll-like receptors (TLR; including TLR-2, TLR-3 and TLR-9), natural killer cell (NKT) agonists, bacterial cell wall products, Lymphocyte Activation Gene-3 (LAG-3), aluminum phosphate, aluminum hydroxide, alum, and saponin.
  • a vaccine adjuvant such as alpha galactose ceramide (o-GC), Toll-like receptors (TLR; including TLR-2, TLR-3 and TLR-9), natural killer cell (NKT) agonists, bacterial cell wall products, Lymphocyte Activation Gene-3 (LAG-3), aluminum phosphate, aluminum hydroxide, alum, and saponin.
  • Adjuvant effects can also be obtained by administering a variety of cytokines along with the immunogenic peptides of the invention including
  • the immunogenic peptides disclosed herein may also be linked directly to, or through, a spacer or linker to: an immunogenic carrier such as serum albumin, tetanus toxoid, keyhole limpet hemocyanin, dextran, or a recombinant virus particle or any synthetic nanoparticles such as liposomes, polymers and metal such as gold nanoparticles; an immunogenic peptide known to stimulate a T helper cell type immune response; a targeting agent such as an antibody or receptor ligand; a stabilising agent such as a lipid; or a conjugate of a plurality of epitopes to a branched lysine core structure, such as the so-called “multiple antigenic peptide" described in (Posnett (1988) J.
  • an immunogenic carrier such as serum albumin, tetanus toxoid, keyhole limpet hemocyanin, dextran, or a recombinant virus particle or any synthetic nanoparticles
  • Useful spacers and linkers are typically comprised of relatively small, neutral molecules, such as amino acids and which are substantially uncharged under physiological conditions. Such spacers are typically selected from the group of nonpolar or neutral polar amino acids, such as glycine, alanine, serine and other similar amino acids. Such optional spacers or linkers need not be comprised of the same residues and thus may be either homo- or heterooligomers. When present, such linkers will commonly be of length at least one or two, commonly 3, 4, 5, 6, and possibly as much as 10 or even up to 20 residues (in the case of amino acids).
  • linkers need not be composed of amino acids but any oligomeric structures will do as well so long as they provide the correct spacing so as to optimise the desired level of immunogenic activity of the immunogenic peptides according to the present invention.
  • the immunogenic peptide may therefore take any form that is capable of eliciting a T cell response.
  • A*02:01 is the most frequently targeted allele with 39 epitopes with the A2 serological antigen associated with another eight responses.
  • A*24:02 is the next most targeted allele, with 17 epitopes (and the A24 serological antigen with an additional epitope).
  • HLA-B is less covered, with only 14 of the 114 restrictions associated with a B allele.
  • HLA B*07:02 was the most targeted B allele with five epitopes.
  • Figure 2 lists the average phenotypic frequencies of each allele. For serological antigens coverage representative of the most common subtype is assumed.
  • population coverage is defined here as the % of individuals that express at least one of the HLA A or B alleles known to restrict the selected epitopes. This analysis reveals that for this set of alleles, there is almost 85% coverage for the general population at the A locus, and about 37% at the B locus. The combined coverage is estimated at over 90% of all individuals. However, these are conservative estimates, since it is likely that any given epitope will be able to be restricted by additional HLA allelic variants closely related to the ones identified and described herein, but that have not yet been investigated.
  • HLA A*02:01 is the most common class I allele in the general worldwide population, and also in most specific populations.
  • A*24:02 is the next most common allele worldwide and is very prevalent in most Asian populations.
  • Other common A alleles are A*01:01, which is prevalent mostly in Caucasians and North Africa, and the A03-supertype alleles A*03:01 and A*ll:01, which are prevalent in Caucasians and Asia, respectively.
  • HLA B is associated with greater polymorphism than the A locus, and as a result only a couple of alleles (B*07:02 and B*08:01) are present with average worldwide frequency >10%. Accordingly, targeting HLA B restricted responses would entail considering a larger number of epitopes and constructs for design.
  • this information highlights a preference for inclusion of at least one epitope restricted by HLA A*02:01 and/or HLA A*24:02 within the immunogenic peptides/vaccine compositions contemplated by the present invention.
  • This approach would also allow for immunogenicity screening in HLA-A*02:01 transgenic mice to support clinical candidate selection.
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three, at least four, at least five or at least six hepatitis B virus (HBV) epitopes selected from the group of epitope sequences derived from Cluster 1 and as set forth in SEQ ID NOs: 12-17, provided that the immunogenic peptide or immunogenic peptide group comprises at least one epitope restricted by human leukocyte antigen (HLA) A*02:01 and/or HLA A*24:02.
  • HLA human leukocyte antigen
  • the required epitope is restricted by HLA A*02:01 and is selected from SEQ ID NO: 12.
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three, at least four, at least five or at least six hepatitis B virus (HBV) epitopes selected from the group of epitope sequences derived from Cluster 3 and as set forth in SEQ ID NOs: 18-23, provided that the immunogenic peptide or immunogenic peptide group comprises at least one epitope restricted by human leukocyte antigen (HLA) A*02:01 and/or HLA A*24:02.
  • HLA human leukocyte antigen
  • the required epitope is restricted by HLA A*02:01 and is selected from SEQ ID NO: 22 and SEQ ID NO: 23.
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three, at least four, at least five or at least six hepatitis B virus (HBV) epitopes selected from the group of epitope sequences derived from Cluster 25 and as set forth in SEQ ID NOs: 24-29, provided that the immunogenic peptide or immunogenic peptide group comprises at least one epitope restricted by human leukocyte antigen (HLA) A*02:01 and/or HLA A*24:02.
  • HLA human leukocyte antigen
  • the required epitope is restricted by HLA A*02:01 and is selected from SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28 and SEQ ID NO: 29.
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three or at least four hepatitis B virus (HBV) epitopes selected from the group of epitope sequences derived from Cluster 8B and as set forth in SEQ ID NOs: 33, 34, 36 and 37, provided that the immunogenic peptide or immunogenic peptide group comprises at least one epitope restricted by human leukocyte antigen (HLA) A*02:01 and/or HLA A*24:02.
  • HLA human leukocyte antigen
  • the required epitope is restricted by HLA A*02:01 and is selected from SEQ ID NO: 36, SEQ ID NO: 37 and SEQ ID NO: 33.
  • the present invention provides an immunogenic peptide or an immunogenic peptide group comprising at least three, at least four, at least five or at least six hepatitis B virus (HBV) epitopes selected from the group of epitope sequences derived from Cluster 13 and as set forth in SEQ ID NOs: 38-43, provided that the immunogenic peptide or immunogenic peptide group comprises at least one epitope restricted by human leukocyte antigen (HLA) A*02:01 and/or HLA A*24:02.
  • HLA human leukocyte antigen
  • the required epitope is restricted by HLA A*02:01 and is selected from SEQ ID NO: 40, SEQ ID NO: 42 and SEQ ID NO: 43.
  • immunogenic peptides described herein are listed in Table 3, below. As described in Example 2 and elsewhere, these sequences are also referred to herein as “clusters” and include multiple overlapping/nested human leukocyte antigen (HLA) class I restricted T-cell epitopes which have been derived from different hepatitis B viral antigens. This includes, without limitation, HBV core protein (“core”), HBV polymerase (“polymerase”), HBV envelope protein (“envelope”) and HBV protein X (“protein X”) virus antigens.
  • HLA human leukocyte antigen
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence LSFLPSDFFPSVRDLLDTASALY (Cluster 1; SEQ ID NO: 1) or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 1.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence YVNTNMGLKILQLLWFHISCLTFGRETVLENL (Cluster 3; SEQ ID NO: 2), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 2.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence SAICSVVRRAFPHCLAFSYMDDVVLGV (Cluster 25; SEQ ID NO: 3), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 3.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in three hepatitis B virus (HBV) epitope cluster sequence QAFTFSPTYKAFLKQQYMNL (Cluster 28; SEQ ID NO: 4), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 4.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence FFLLTRILTIPQSLDSWWTSLNFLGGAPTCP (Cluster 8; SEQ ID NO: 5), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 5.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence FLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFL (Cluster 8B; SEQ ID NO: 6), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 6.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence SVRFSWLSLLVPFVQWFVGLSPTVWLSV (Cluster 13; SEQ ID NO: 7), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 7.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence VCWGELMTLATWVGSNLEDPA (Cluster 2; SEQ ID NO: 8), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 8.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence LSTLPETTVVRRRGRSPRRRTPSPRRRRSQ (Cluster 5; SEQ ID NO: 9), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 9.
  • HBV hepatitis B virus
  • the present invention provides an immunogenic peptide comprising or consisting in the hepatitis B virus (HBV) epitope cluster sequence FLPSDFFPSVRDLLDTASALY (Cluster IB; SEQ ID NO: 10), or a variant sequence which shares at least 80% sequence identity to SEQ ID NO: 10.
  • HBV hepatitis B virus
  • able 3 Immunogenic 'Cluster' Peptides
  • Applicants assembled discrete epitopes into 20-35 residue immunogenic peptides in which each epitope was separated by synthetic or naturally occurring spacer residue(s). Applicants refer to these peptide constructs as "beads".
  • the number of spacer residues may be (e.g.) one, two, three, four or five amino acid residues, although either one or two synthetic or naturally occurring spacer residue(s) between each epitope is preferred. Further information in relation to the bead constructs contemplated by the present invention is presented in Example 4 which follows.
  • Linkers such as amino acid or peptidomimetic sequences, may be inserted between the epitope sequences and the addition (e.g., heterologous functional domain) so that the two entities maintain, at least in part, a distinct function or activity.
  • Linkers may have one or more properties that may include a flexible conformation, an inability to form an ordered secondary structure or a hydrophobic or charged character, which could promote or interact with either domain.
  • Amino acids typically found in flexible protein regions include Gly, Asn and Ser. Other near neutral amino acids, such as Thr and Ala, may also be used in the linker sequence.
  • an immunogenic peptide comprising three hepatitis B virus (HBV) epitopes selected from any one of SEQ ID Nos 12-97, or a variant sequence which shares at least 75%, 80%, 85%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 12-97, wherein a spacer comprising one, two, three, four or more amino acid residues is located between each HBV epitope and wherein the immunogenic peptide comprises at least one HBV epitope restricted by human leukocyte antigen (HLA) A*02:01.
  • HLA human leukocyte antigen
  • an immunogenic peptide comprising three hepatitis B virus (HBV) epitopes selected from SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 43, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 55, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 79, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 97, or a variant sequence which shares at least 75%, 80%, 85%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or
  • an immunogenic peptide comprising hepatitis B virus (HBV) epitopes defined by SEQ ID Nos: 12, 48 and 50, wherein a spacer comprising one, two, three, four or more amino acid residues is located between each HBV epitope.
  • HBV hepatitis B virus
  • an immunogenic peptide comprising hepatitis B virus (HBV) epitopes defined by SEQ ID Nos: 13, 33 and 92, wherein a spacer comprising one, two, three, four or more amino acid residues is located between each HBV epitope.
  • HBV hepatitis B virus
  • an immunogenic peptide comprising hepatitis B virus (HBV) epitopes defined by SEQ ID Nos: 40, 74 and 91, wherein a spacer comprising one, two, three, four or more amino acid residues is located between each HBV epitope.
  • HBV hepatitis B virus
  • an immunogenic peptide comprising hepatitis B virus (HBV) epitopes defined by SEQ ID Nos: 34, 43 and 55, wherein a spacer comprising one, two, three, four or more amino acid residues is located between each HBV epitope.
  • HBV hepatitis B virus
  • an immunogenic peptide comprising hepatitis B virus (HBV) epitopes defined by SEQ ID Nos: 39, 71 and 75, wherein a spacer comprising one, two, three, four or more amino acid residues is located between each HBV epitope.
  • HBV hepatitis B virus
  • an immunogenic peptide comprising hepatitis B virus (HBV) epitopes defined by SEQ ID Nos: 15, 72 and 97, wherein a spacer comprising one, two, three, four or more amino acid residues is located between each HBV epitope.
  • HBV hepatitis B virus
  • an immunogenic peptide comprising hepatitis B virus (HBV) epitopes defined by SEQ ID Nos: 18, 37 and 73, wherein a spacer comprising one, two, three, four or more amino acid residues is located between each HBV epitope.
  • HBV hepatitis B virus
  • an immunogenic peptide comprising hepatitis B virus (HBV) epitopes defined by SEQ ID Nos: 20, 52 and 79, wherein a spacer comprising one, two, three, four or more amino acid residues is located between each HBV epitope.
  • HBV hepatitis B virus
  • an immunogenic peptide comprising hepatitis B virus (HBV) epitopes defined by SEQ ID Nos: 12, 79 and 20, wherein a spacer comprising one, two, three, four or more amino acid residues is located between each HBV epitope.
  • HBV hepatitis B virus
  • an immunogenic peptide comprising hepatitis B virus (HBV) epitopes defined by SEQ ID Nos: 18, 33 and 37, wherein a spacer comprising one, two, three, four or more amino acid residues is located between each HBV epitope.
  • an immunogenic peptide comprising hepatitis B virus (HBV) epitopes defined by SEQ ID Nos: 15, 40 and 97, wherein a spacer comprising one, two, three, four or more amino acid residues is located between each HBV epitope.
  • an immunogenic peptide comprising hepatitis B virus (HBV) epitopes defined by SEQ ID Nos: 39, 43 and 75, wherein a spacer comprising one, two, three, four or more amino acid residues is located between each HBV epitope.
  • HBV hepatitis B virus
  • an immunogenic peptide comprising hepatitis B virus (HBV) epitopes defined by SEQ ID Nos: 34, 55 and 71, wherein a spacer comprising one, two, three, four or more amino acid residues is located between each HBV epitope.
  • HBV hepatitis B virus
  • an immunogenic peptide comprising hepatitis B virus (HBV) epitopes defined by SEQ ID Nos: 72, 74 and 91, wherein a spacer comprising one, two, three, four or more amino acid residues is located between each HBV epitope.
  • HBV hepatitis B virus
  • an immunogenic peptide comprising hepatitis B virus (HBV) epitopes defined by SEQ ID Nos: 13, 73 and 92, wherein a spacer comprising one, two, three, four or more amino acid residues is located between each HBV epitope.
  • HBV hepatitis B virus
  • an immunogenic peptide comprising hepatitis B virus (HBV) epitopes defined by SEQ ID Nos: 48, 50 and 52, wherein a spacer comprising one, two, three, four or more amino acid residues is located between each HBV epitope.
  • HBV hepatitis B virus
  • the presence of three HBV epitopes requires the presence of two discrete amino acid spacers comprising one, two, three, four or more amino acid residues.
  • each spacer comprises two amino acid residues.
  • each spacer comprises two alanine residues (i.e.) 5'-AA-3' (SEQ ID NO: 169).
  • the present invention provides an immunogenic peptide comprising or consisting in FLPSDFFPSVAASTLPETTVVRRAAEYLVSFGVW (Bead 1; SEQ ID NO: 111), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 111.
  • the present invention provides an immunogenic peptide comprising or consisting in FLLTRILTIAALPSDFFPSVAACLFKDWEEL (Bead 2; SEQ ID NO: 112), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 112.
  • the present invention provides an immunogenic peptide comprising or consisting in WLSLLVPFVAAVLHKRTLGLAAKYTSFPWLL (Bead 3; SEQ ID NO:
  • the present invention provides an immunogenic peptide comprising or consisting in GLSPTVWLSVAASWWTSLNFLAALLCLIFLLV (Bead 4; SEQ ID NO:
  • the present invention provides an immunogenic peptide comprising or consisting in GLSRYVARLAASWLSLLVPFAAHTLWKAGILYK (Bead 5; SEQ ID NO:
  • the present invention provides an immunogenic peptide comprising or consisting in FLLSLGIHLAAFFPSIRDLLAAVLCLRPVGA (Bead 6; SEQ ID NO: 116), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 116.
  • the present invention provides an immunogenic peptide comprising or consisting in SLYADSPSVAAVLQAGFFLLAAYVNTNMGLK (Bead 7; SEQ ID NO:
  • the present invention provides an immunogenic peptide comprising or consisting in LLDYQGMLPVAALWFHISCLTFAATPARVTGGVF (Bead 8; SEQ ID NO:
  • the present invention provides an immunogenic peptide comprising or consisting in FLPSDFFPSVAATPARVTGGVFAALWFHISCLTF (Bead 9; SEQ ID NO:
  • the present invention provides an immunogenic peptide comprising or consisting in FLLTRILTIAAYVNTNMGLKAAVLQAGFFLL (Bead 10; SEQ ID NO:
  • the present invention provides an immunogenic peptide comprising or consisting in WLSLLVPFVAAVLCLRPVGAAAFFPSIRDLL (Bead 11; SEQ ID NO:
  • the present invention provides an immunogenic peptide comprising or consisting in GLSPTVWLSVAAHTLWKAGILYKAASWLSLLVPF (Bead 12; SEQ ID NO: 122), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 122.
  • the present invention provides an immunogenic peptide comprising or consisting in GLSRYVARLAALLCLIFLLVAASWWTSLNFL (Bead 13; SEQ ID NO:
  • the present invention provides an immunogenic peptide comprising or consisting in FLLSLGIHLAAKYTSFPWLLAAVLHKRTLGL (Bead 14; SEQ ID NO:
  • the present invention provides an immunogenic peptide comprising or consisting in SLYADSPSVAACLFKDWEELAALPSDFFPSV (Bead 15; SEQ ID NO:
  • the present invention provides an immunogenic peptide comprising or consisting in LLDYQGMLPVAASTLPETTVVRRAAEYLVSFGVW (Bead 16; SEQ ID NO: 126), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 126.
  • each spacer comprises one, two, three, four or more naturally occurring amino acid residues located immediately adjacent to the 5' or 3'-end of the HBV epitope in the human HBV protein reference sequence (e.g. human HBV envelope protein, human HBV capsid protein, human HBV protein P, human HBV protein X etc).
  • human HBV protein reference sequence e.g. human HBV envelope protein, human HBV capsid protein, human HBV protein P, human HBV protein X etc.
  • the amino acid sequences associated with human HBV proteins are readily available via public access databases (e.g.) protein database available at the National Library of Medicine.
  • each spacer in the immunogenic peptide as defined comprises two naturally occurring amino acid residues located immediately adjacent to the 5' or 3'-end of the HBV epitope in the human HBV protein reference sequence (e.g. human HBV envelope protein, human HBV capsid protein, human HBV protein P, human HBV protein X etc).
  • human HBV envelope protein e.g. human HBV capsid protein, human HBV protein P, human HBV protein X etc.
  • this includes the di-amino acid spacers selected from 5'-RL-3' (SEQ ID NO: 170), 5'-PF-3' (SEQ ID NO: 171), 5'-RD-3' (SEQ ID NO: 172), 5'-QK-3' (SEQ ID NO: 173), 5'-SR-3' (SEQ ID NO: 174), 5'-ID-3' (SEQ ID NO: 175), 5'-GL-3' (SEQ ID NO: 176), 5'-SF-3' (SEQ ID NO: 177), 5'-VL-3' (SEQ ID NO: 178), 5'- ND-3' (SEQ ID NO: 179), 5'-DD-3' (SEQ ID NO: 180), 5'-PL-3' (SEQ ID NO: 181), 5'-TS-3' (SEQ ID NO: 182), 5'-CL-3' (SEQ ID NO: 183), 5'-GR-3' (SEQ ID NO: 184), 5'-RR
  • the present invention provides an immunogenic peptide comprising or consisting in FLPSDFFPSVRLSTLPETTVVRRRLEYLVSFGVW (Bead 17; SEQ ID NO: 127), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 127.
  • the present invention provides an immunogenic peptide comprising or consisting in FLLTRILTIPFLPSDFFPSVRDCLFKDWEEL (Bead 18; SEQ ID NO: 128), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 128.
  • the present invention provides an immunogenic peptide comprising or consisting in WLSLLVPFVQKVLHKRTLGLSRKYTSFPWLL (Bead 19; SEQ ID NO: 129), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 129.
  • the present invention provides an immunogenic peptide comprising or consisting in GLSPTVWLSVIDSWWTSLNFLGLLLCLIFLLV (Bead 20; SEQ ID NO: 130), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 130.
  • the present invention provides an immunogenic peptide comprising or consisting in GLSRYVARLSFSWLSLLVPFVLHTLWKAGILYK (Bead 21; SEQ ID NO: 131), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 131.
  • the present invention provides an immunogenic peptide comprising or consisting in FLLSLGIHLNDFFPSIRDLLDDVLCLRPVGA (Bead 22; SEQ ID NO: 132), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 132.
  • the present invention provides an immunogenic peptide comprising or consisting in SLYADSPSVPLVLQAGFFLLTSYVNTNMGLK (Bead 23; SEQ ID NO: 133), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 133.
  • the present invention provides an immunogenic peptide comprising or consisting in LLDYQGMLPVCLLWFHISCLTFGRTPARVTGGVF (Bead 24; SEQ ID NO: 134), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 134.
  • the present invention provides an immunogenic peptide comprising or consisting in FLPSDFFPSVRRTPARVTGGVFLLLWFHISCLTF (Bead 25; SEQ ID NO: 135), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 135.
  • the present invention provides an immunogenic peptide comprising or consisting in FLLTRILTIPSYVNTNMGLKILVLQAGFFLL (Bead 26; SEQ ID NO: 136), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 136.
  • the present invention provides an immunogenic peptide comprising or consisting in WLSLLVPFVQDVLCLRPVGAEDFFPSIRDLL (Bead 27; SEQ ID NO: 137), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 137.
  • the present invention provides an immunogenic peptide comprising or consisting in GLSPTVWLSVILHTLWKAGILYKRFSWLSLLVPF (Bead 28; SEQ ID NO: 138), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 138.
  • the present invention provides an immunogenic peptide comprising or consisting in GLSRYVARLSLLLCLIFLLVLDSWWTSLNFL (Bead 29; SEQ ID NO: 139), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 139.
  • the present invention provides an immunogenic peptide comprising or consisting in FLLSLGIHLNRKYTSFPWLLGKVLHKRTLGL (Bead 30; SEQ ID NO: 140), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 140.
  • the present invention provides an immunogenic peptide comprising or consisting in SLYADSPSVPDCLFKDWEELGFLPSDFFPSV (Bead 31; SEQ ID NO: 141), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 141.
  • the present invention provides an immunogenic peptide comprising or consisting in LLDYQGMLPVCLSTLPETTVVRRRLEYLVSFGVW (Bead 32; SEQ ID NO: 142), or a variant sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 142.
  • able 4A Bead Constructs 1-8; Dominant Beads with Alanine-Alanine Spacers
  • each peptide cluster construct assessed the potential for each peptide cluster construct to recall HBV-specific class II restricted T cells, and class II restricted T cells in general. This analysis reveals that six of the cluster constructs were associated with known HBV-derived HTL epitopes, with some of the cluster constructs containing multiple epitopes. HLA binding predictions for a panel of the most common HLA class II alleles worldwide demonstrated that each construct is associated with about a dozen or more, an average of 110, and up to 283, different possible peptide/HLA class II binding combinations, which have the potential to provide construct specific helper responses.
  • HLA class II The coincidental coverage of HLA class II varies amongst different constructs, thus providing additional potential criteria to select and optimize epitope selection.
  • the expected population coverage for the entire set of 86 epitopes is 90.2%.
  • the "Top 8 clusters" in terms of population coverage at the time of analysis i.e. Cluster 3 (SEQ ID NO: 2), Cluster 25 (SEQ ID NO: 3), Cluster 28 (SEQ ID NO: 4), Cluster 8 (SEQ ID NO: 5), Cluster 13 (SEQ ID NO: 7), Cluster 2 (SEQ ID NO: 8), Cluster 5 (SEQ ID NO: 9) and Cluster IB (SEQ ID NO: 10); 29 unique epitopes) achieved 63.7% of total responses with a population coverage of 89.8%.
  • the "Top 6 clusters" at the time of analysis i.e.
  • Cluster 3 (SEQ ID NO: 2), Cluster 25 (SEQ ID NO: 3), Cluster 28 (SEQ ID NO: 4), Cluster 8 (SEQ ID NO: 5), Cluster 5 (SEQ ID NO: 9) and Cluster IB (SEQ ID NO: 10)) while having slightly fewer responses achieved a comparable level of population coverage with 88.5%.
  • Table 5 in Example 2.
  • both 'sets' of clusters achieved a very similar level of population coverage (i.e.) 89.8% and 88.5% versus 90.2%.
  • the level of population coverage i.e. by geographical area and/or by ethnicity
  • Cluster 1 and Cluster 8B is expected to improve if similar calculations were run using subsequently developed clusters, namely Cluster 1 and Cluster 8B.
  • the cluster/bead peptide sequences defined by SEQ ID Nos: 1-11 include multiple (and in the case of clusters nested/overlapping) human leukocyte antigen (HLA) class I restricted T-cell epitopes that are derived from different hepatitis B viral antigens.
  • HLA human leukocyte antigen
  • the present invention provides a vaccine composition
  • a vaccine composition comprising two or more immunogenic peptides (or immunogenic peptide compositions) as described herein together with a pharmacologically acceptable carrier or vaccine delivery vehicle, provided that at least two immunogenic peptides comprise a cluster of T-cell epitopes derived from different virus antigens.
  • the vaccine composition comprises (i) an immunogenic peptide which includes T-cell epitopes derived from HBV capsid/external core protein and (ii) an immunogenic peptide which includes T-cell epitopes derived from HBV envelope protein together with a pharmacologically acceptable carrier or vaccine delivery vehicle.
  • the vaccine composition comprises (i) an immunogenic peptide which includes T-cell epitopes derived from HBV capsid/external core protein and (ii) an immunogenic peptide which includes T-cell epitopes derived from HBV protein P together with a pharmacologically acceptable carrier or vaccine delivery vehicle.
  • the vaccine composition comprises (i) an immunogenic peptide which includes T-cell epitopes derived from HBV capsid/external core protein and (ii) an immunogenic peptide which includes T-cell epitopes derived from HBV protein X together with a pharmacologically acceptable carrier or vaccine delivery vehicle.
  • the vaccine composition comprises (i) an immunogenic peptide which includes T-cell epitopes derived from HBV capsid/external core protein and (ii) an immunogenic peptide which includes T-cell epitopes derived from HBV polymerase together with a pharmacologically acceptable carrier or vaccine delivery vehicle.
  • the vaccine composition comprises (i) an immunogenic peptide which includes T-cell epitopes derived from HBV envelope protein and (ii) an immunogenic peptide which includes T-cell epitopes derived from HBV protein P together with a pharmacologically acceptable carrier or vaccine delivery vehicle.
  • the vaccine composition comprises (i) an immunogenic peptide which includes T-cell epitopes derived from HBV envelope protein and (ii) an immunogenic peptide which includes T-cell epitopes derived from HBV protein X together with a pharmacologically acceptable carrier or vaccine delivery vehicle.
  • the vaccine composition comprises (i) an immunogenic peptide which includes T-cell epitopes derived from HBV envelope protein and (ii) an immunogenic peptide which includes T-cell epitopes derived from HBV polymerase together with a pharmacologically acceptable carrier or vaccine delivery vehicle.
  • the vaccine composition comprises (i) an immunogenic peptide which includes T-cell epitopes derived from HBV protein P and (ii) an immunogenic peptide which includes T-cell epitopes derived from HBV protein X together with a pharmacologically acceptable carrier or vaccine delivery vehicle.
  • the vaccine composition comprises (i) an immunogenic peptide which includes T-cell epitopes derived from HBV protein P and (ii) an immunogenic peptide which includes T-cell epitopes derived from HBV polymerase together with a pharmacologically acceptable carrier or vaccine delivery vehicle.
  • the vaccine composition comprises (i) an immunogenic peptide which includes T-cell epitopes derived from HBV protein X and (ii) an immunogenic peptide which includes T-cell epitopes derived from HBV polymerase together with a pharmacologically acceptable carrier or vaccine delivery vehicle.
  • the vaccine composition comprises (i) an immunogenic peptide which includes T-cell epitopes derived from HBV capsid/external core protein, (ii) an immunogenic peptide which includes T-cell epitopes derived from HBV envelope protein and (iii) an immunogenic peptide which includes T-cell epitopes derived from HBV protein B, and optionally a pharmacologically acceptable carrier or vaccine delivery vehicle.
  • the vaccine/ compositions of the present invention comprise at least two or at least three immunogenic peptides selected from the following immunogenic peptide combinations:
  • SEQ ID NO 7 (Cluster 13) and SEQ ID NO: 10 (Cluster IB); SEQ ID NO: 7 (Cluster 13) and SEQ ID NO: 11 (Bead 17);
  • SEQ ID NO: 10 Cluster IB
  • SEQ ID NO: 11 Bead 17
  • SEQ ID NO: 1 (Cluster 1), SEQ ID NO: 2 (Cluster 3), SEQ ID NO: 3 (Cluster 25);
  • SEQ ID NO: 1 (Cluster 1), SEQ ID NO: 2 (Cluster 3), SEQ ID NO: 4 (Cluster 28);
  • SEQ ID NO 1 (Cluster 1), SEQ ID NO 2 (Cluster 3), SEQ ID NO: 5 (Cluster 8);
  • SEQ ID NO 1 (Cluster 1), SEQ ID NO 2 (Cluster 3), SEQ ID NO: 6 (Cluster 8B);
  • SEQ ID NO 1 (Cluster 1), SEQ ID NO 2 (Cluster 3), SEQ ID NO: 7 (Cluster 13);
  • SEQ ID NO 1 (Cluster 1), SEQ ID NO 2 (Cluster 3), SEQ ID NO: 9 (Cluster 5);
  • SEQ ID NO 1 (Cluster 1), SEQ ID NO 2 (Cluster 3), SEQ ID NO: 10 (Cluster IB);
  • SEQ ID NO 1 (Cluster 1), SEQ ID NO 2 (Cluster 3), SEQ ID NO: 11 (Bead 17);
  • SEQ ID NO 1 (Cluster 1), SEQ ID NO 5 (Cluster 8), SEQ ID NO: 6 (Cluster 8B);
  • SEQ ID NO 1 (Cluster 1), SEQ ID NO 5 (Cluster 8), SEQ ID NO: 7 (Cluster 13);
  • SEQ ID NO 1 (Cluster 1), SEQ ID NO 5 (Cluster 8), SEQ ID NO: 8 (Cluster 2);
  • SEQ ID NO 1 (Cluster 1), SEQ ID NO 5 (Cluster 8), SEQ ID NO: 9 (Cluster 5);
  • SEQ ID NO 1 (Cluster 1), SEQ ID NO 5 (Cluster 8), SEQ ID NO: 10 (Cluster IB);
  • SEQ ID NO 1 (Cluster 1), SEQ ID NO 6 (Cluster 8B), SEQ ID NO: 7 (Cluster 13); SEQ ID NO 1 (Cluster 1), SEQ ID NO 6 (Cluster 8B), SEQ ID NO: 8 (Cluster 2);
  • SEQ ID NO 1 (Cluster 1), SEQ ID NO 6 (Cluster 8B), SEQ ID NO: 9 (Cluster 5);
  • SEQ ID NO 1 (Cluster 1), SEQ ID NO 6 (Cluster 8B), SEQ ID NO: 10 (Cluster IB);
  • SEQ ID NO 1 (Cluster 1), SEQ ID NO 6 (Cluster 8B), SEQ ID NO: 11 (Bead 17);
  • SEQ ID NO 1 (Cluster 1), SEQ ID NO 7 (Cluster 13), SEQ ID NO: 8 (Cluster 2);
  • SEQ ID NO 1 (Cluster 1), SEQ ID NO 7 (Cluster 13), SEQ ID NO: 9 (Cluster 5);
  • SEQ ID NO 1 (Cluster 1), SEQ ID NO 7 (Cluster 13), SEQ ID NO: 10 (Cluster IB);
  • SEQ ID NO 1 (Cluster 1), SEQ ID NO 7 (Cluster 13), SEQ ID NO: 11 (Bead 17);
  • SEQ ID NO 1 (Cluster 1), SEQ ID NO 9 (Cluster 5), SEQ ID NO: 10 (Cluster IB);
  • SEQ ID NO 1 (Cluster 1), SEQ ID NO 9 (Cluster 5), SEQ ID NO: 11 (Bead 17);
  • SEQ ID NO 1 (Cluster 1), SEQ ID NO 10 (Cluster IB), SEQ ID NO: 11 (Bead 17);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 3 (Cluster 25), SEQ ID NO: 4 (Cluster 28);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 3 (Cluster 25), SEQ ID NO: 5 (Cluster 8);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 3 (Cluster 25), SEQ ID NO: 6 (Cluster 8B);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 3 (Cluster 25), SEQ ID NO: 7 (Cluster 13);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 3 (Cluster 25), SEQ ID NO: 8 (Cluster 2);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 3 (Cluster 25), SEQ ID NO: 9 (Cluster 5);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 3 (Cluster 25), SEQ ID NO: 10 (Cluster IB);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 3 (Cluster 25), SEQ ID NO: 11 (Bead 17);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 4 (Cluster 28), SEQ ID NO: 5 (Cluster 8);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 4 (Cluster 28), SEQ ID NO: 6 (Cluster 8B);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 4 (Cluster 28), SEQ ID NO: 7 (Cluster 13);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 4 (Cluster 28), SEQ ID NO: 8 (Cluster 2);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 4 (Cluster 28), SEQ ID NO: 9 (Cluster 5);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 4 (Cluster 28), SEQ ID NO: 10 (Cluster IB);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 4 (Cluster 28), SEQ ID NO: 11 (Bead 17);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 5 (Cluster 8), SEQ ID NO: 6 (Cluster 8B);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 5 (Cluster 8), SEQ ID NO: 7 (Cluster 13);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 5 (Cluster 8), SEQ ID NO: 8 (Cluster 2);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 5 (Cluster 8), SEQ ID NO: 9 (Cluster 5);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 5 (Cluster 8), SEQ ID NO: 10 (Cluster IB);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 5 (Cluster 8), SEQ ID NO: 11 (Bead 17);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 6 (Cluster 8B), SEQ ID NO: 7 (Cluster 13);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 6 (Cluster 8B), SEQ ID NO: 8 (Cluster 2);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 6 (Cluster 8B), SEQ ID NO: 9 (Cluster 5); SEQ ID NO 2 (Cluster 3), SEQ ID NO 6 (Cluster 8B), SEQ ID NO: 10 (Cluster IB);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 6 (Cluster 8B), SEQ ID NO: 11 (Bead 17);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 7 (Cluster 13), SEQ ID NO: 8 (Cluster 2);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 7 (Cluster 13), SEQ ID NO: 9 (Cluster 5);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 7 (Cluster 13), SEQ ID NO: 10 (Cluster IB);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 7 (Cluster 13), SEQ ID NO: 11 (Bead 17);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 8 (Cluster 2), SEQ ID NO: 9 (Cluster 5);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 8 (Cluster 2), SEQ ID NO: 10 (Cluster IB);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 8 (Cluster 2), SEQ ID NO: 11 (Bead 17);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 9 (Cluster 5), SEQ ID NO: 10 (Cluster IB);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 9 (Cluster 5), SEQ ID NO: 11 (Bead 17);
  • SEQ ID NO 2 (Cluster 3), SEQ ID NO 10 (Cluster IB), SEQ ID NO: 11 (Bead 17);
  • SEQ ID NO 3 (Cluster 25), SEQ ID NO 4 (Cluster 28), SEQ ID NO: 6 (Cluster 8B);
  • SEQ ID NO 3 (Cluster 25), SEQ ID NO 4 (Cluster 28), SEQ ID NO: 7 (Cluster 13);
  • SEQ ID NO 3 (Cluster 25), SEQ ID NO 4 (Cluster 28), SEQ ID NO: 8 (Cluster 2);
  • SEQ ID NO 3 (Cluster 25), SEQ ID NO 4 (Cluster 28), SEQ ID NO: 10 (Cluster IB);
  • SEQ ID NO 3 (Cluster 25), SEQ ID NO 4 (Cluster 28), SEQ ID NO: 11 (Bead 17);
  • SEQ ID NO 3 (Cluster 25), SEQ ID NO 5 (Cluster 8), SEQ ID NO: 6 (Cluster 8B);
  • SEQ ID NO 3 (Cluster 25), SEQ ID NO 5 (Cluster 8), SEQ ID NO: 7 (Cluster 13);
  • SEQ ID NO 3 (Cluster 25), SEQ ID NO 5 (Cluster 8), SEQ ID NO: 8 (Cluster 2);
  • SEQ ID NO 3 (Cluster 25), SEQ ID NO 5 (Cluster 8), SEQ ID NO: 10 (Cluster IB);
  • SEQ ID NO 3 (Cluster 25), SEQ ID NO 5 (Cluster 8), SEQ ID NO: 11 (Bead 17);
  • SEQ ID NO 3 (Cluster 25), SEQ ID NO 6 (Cluster 8B), SEQ ID NO: 7 (Cluster 13);
  • SEQ ID NO 3 (Cluster 25), SEQ ID NO 6 (Cluster 8B), SEQ ID NO: 8 (Cluster 2);
  • SEQ ID NO 3 (Cluster 25), SEQ ID NO 6 (Cluster 8B), SEQ ID NO: 9 (Cluster 5);
  • SEQ ID NO: 3 (Cluster 25), SEQ ID NO: 6 (Cluster 8B), SEQ ID NO: 11 (Bead 17); SEQ ID NO: 3 (Cluster 25), SEQ ID NO: 7 (Cluster 13), SEQ ID NO: 8 (Cluster 2); SEQ ID NO: 3 (Cluster 25), SEQ ID NO: 7 (Cluster 13), SEQ ID NO: 9 (Cluster 5); SEQ ID NO: 3 (Cluster 25), SEQ ID NO: 7 (Cluster 13), SEQ ID NO: 10 (Cluster IB) SEQ ID NO: 3 (Cluster 25), SEQ ID NO: 7 (Cluster 13), SEQ ID NO: 11 (Bead 17); SEQ ID NO: 3 (Cluster 25), SEQ ID NO: 8 (Cluster 2), SEQ ID NO: 9 (Cluster 5); SEQ ID NO: 3 (Cluster 25), SEQ ID NO: 8 (Cluster 2), SEQ ID NO: 10 (Cluster IB); SEQ ID NO: 3 (Cluster 25), SEQ ID NO: 8 (
  • SEQ ID NO: 5 (Cluster 8), SEQ ID NO: 10 (Cluster IB), SEQ ID NO: 11 (Bead 17);
  • SEQ ID NO: 6 (Cluster 8B), SEQ ID NO: 7 (Cluster 13), SEQ ID NO: 8 (Cluster 2);
  • SEQ ID NO: 6 (Cluster 8B), SEQ ID NO: 7 (Cluster 13), SEQ ID NO: 9 (Cluster 5);
  • SEQ ID NO 6 (Cluster 8B), SEQ ID NO: 7 (Cluster 13), SEQ ID NO: 11 (Bead 17);
  • SEQ ID NO 6 (Cluster 8B), SEQ ID NO: 8 (Cluster 2), SEQ ID NO: 9 (Cluster 5);
  • SEQ ID NO 6 (Cluster 8B), SEQ ID NO: 8 (Cluster 2), SEQ ID NO: 10 (Cluster IB);
  • SEQ ID NO 6 (Cluster 8B), SEQ ID NO: 8 (Cluster 2), SEQ ID NO: 11 (Bead 17);
  • SEQ ID NO 6 (Cluster 8B), SEQ ID NO: 9 (Cluster 5), SEQ ID NO: 10 (Cluster IB);
  • SEQ ID NO 6 (Cluster 8B), SEQ ID NO: 9 (Cluster 5), SEQ ID NO: 11 (Bead 17);
  • SEQ ID NO 6 (Cluster 8B), SEQ ID NO: 10 (Cluster IB), SEQ ID NO: 11 (Bead 17);
  • SEQ ID NO 7 (Cluster 13), SEQ ID NO: 8 (Cluster 2), SEQ ID NO: 9 (Cluster 5);
  • SEQ ID NO 7 (Cluster 13), SEQ ID NO: 8 (Cluster 2), SEQ ID NO: 10 (Cluster IB);
  • SEQ ID NO 7 (Cluster 13), SEQ ID NO: 8 (Cluster 2), SEQ ID NO: 11 (Bead 17);
  • SEQ ID NO 7 (Cluster 13), SEQ ID NO: 9 (Cluster 5), SEQ ID NO: 10 (Cluster IB);
  • SEQ ID NO 7 (Cluster 13), SEQ ID NO: 9 (Cluster 5), SEQ ID NO: 11 (Bead 17);
  • SEQ ID NO 7 (Cluster 13), SEQ ID NO: 10 (Cluster IB), SEQ ID NO: 11 (Bead 17);
  • SEQ ID NO 8 (Cluster 2), SEQ ID NO: 9 (Cluster 5), SEQ ID NO: 10 (Cluster IB);
  • SEQ ID NO 8 (Cluster 2), SEQ ID NO: 9 (Cluster 5), SEQ ID NO: 11 (Bead 17);
  • SEQ ID NO 8 (Cluster 2), SEQ ID NO: 10 (Cluster IB), SEQ ID NO: 11 (Bead 17);
  • SEQ ID NO 9 (Cluster 5), SEQ ID NO: 10 (Cluster IB), SEQ ID NO: 11 (Bead 17); or any immunogenic variant peptide sequence which shares at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to any one of SEQ ID Nos: 1-11 based on the preceding combinations.
  • the vaccine/compositions described herein may include (e.g.) combinations comprising more than three immunogenic peptides, for example, four, five, six, seven or more immunogenic peptides of the present invention.
  • the requirement to formulate multiple immunogenic peptides in a vaccine composition (or indeed combine vaccine compositions) will largely depend on phenotypic frequency and depth of coverage necessary to achieve the level of desired population coverage.
  • compositions including vaccine compositions, according to the present invention may be in the form of a composition comprising two or more immunogenic peptides and wherein each immunogenic peptide is present in any desired relative abundance.
  • Such compositions can be homogeneous or heterogeneous with respect to the individual immunogenic peptides of the invention.
  • the immunogenic peptides according to the present invention may be formulated separately in the vaccine/compositions described herein, or the immunogenic peptides may be linked directly together, or through a spacer or linker, to form a larger structure, such as polymeric structure.
  • the immunogenic peptides may therefore be linked by any and all means that can be devised by the chemist so long as the immunogenic activity of the overall structure or complex is maintained or, at least, not reduced below a level useful for the methods of the invention (i.e., especially where said immunogenic activity comprises being capable of eliciting a cytotoxic T cell response).
  • Example 3 demonstrates that in Caucasian populations, which corresponds to the prevalent ethnicity of patients in which the epitopes were defined, coverage for the peptide cluster constructs in the 59.4 to 80.2% range was calculated, with the average number of epitopes recognized in the 0.64 to 1.95 range. Combination of the top three cluster constructs was associated with 95.9% coverage, and an average of 3.94 epitopes recognised.
  • Example 7 describes the generation of population specific cluster comprising an immunodominant HLA-C*08:01 restricted T cell epitope in the context of a cohort of HBV infected subjects in Southeast Asia recently reported by Tan et al. (PMID 24227846).
  • the HBV envelope 182-191 epitope can be used to generate a new 35-mer cluster, representing a frameshift of Cluster 8; Applicants have denominated this frameshift Cluster 8B.
  • This new cluster represents a slight improvement over Cluster 8 in terms of total positive responses and population coverage worldwide and in Caucasians. However, and importantly, coverage of Southeast Asia is predicted to be 68%, compared to 59% for Cluster 8, making it one of the stronger clusters for covering that population.
  • Cluster 8B nest three different known promiscuous HTL epitopes, and is associated with the most predicted HLA class II binding events.
  • the present invention further contemplates immunogenic peptides and vaccine compositions useful for the treatment of chronic hepatitis B virus infection that achieves a minimum/desired population coverage, based on geography and/or for a target population, with coverage of at least 50%, or at least 55%, or at least 60%, at least 65%, or at least 70%, or at least 75%, or at least 80%, or at least 81%, or at least 82%, or at least 83%, or at least 84%, or at least 85%, or at least 86%, or at least 87%, or at least 88%, or at least 89%, or at least 90%, or at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or higher population coverage.
  • immunogenic peptide In certain examples the immunogenic peptide, immunogenic peptide groups, and vaccine compositions described herein achieve:
  • the immunogenic peptide, immunogenic peptide group, and vaccine compositions described herein achieve a population coverage of at least 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher in a specific population group selected from Europe, North Africa, North-East Asia, South-East Asia, South-West Asia, Hispanic North and South America, American Indian, Sub-Saharan and South Pacific.
  • the immunogenic peptide, immunogenic peptide group, and vaccine compositions described herein achieve a population coverage of at least 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher in a greater China region including, but not limited to, China, Hong Kong, Macau and Taiwan.
  • FIG. 5 demonstrates the immunogenicity of cluster and bead constructs in a peripheral blood mononuclear cell (PBMC) assay using blood samples taken from healthy controls, as well as acute and chronic HBV affected patients.
  • PBMC peripheral blood mononuclear cell
  • the immunogenic peptides of the invention may be utilised for purpose of preventing, suppressing or treating HBV infection, such as where the antigen is being expressed by HBV infected cells.
  • prevention relates to a process of prophylaxis in which an animal, especially a mammal, and most especially a human, is exposed to an immunogen of the present invention prior to the induction or onset of the disease process.
  • the immunogenic peptide(s) could be administered to the general population as is frequently done for infectious diseases.
  • the term “suppression” is often used to describe a condition wherein the disease process has already begun but obvious symptoms of said condition have yet to be realised.
  • the cells of an individual may have become infected but no outside signs of the disease have yet been clinically recognised.
  • the term prophylaxis can be applied to encompass both prevention and suppression.
  • the term “treatment” is often utilised to mean the clinical application of agents to combat an already existing condition whose clinical presentation has already been realised in a patient. This would occur where an individual has already been diagnosed as having an HBV infection.
  • the present invention provides a method for treating hepatitis B virus (HBV) infection in a human subject, the method comprising administering to the human subject at least one vaccine composition as described herein.
  • HBV hepatitis B virus
  • the present invention further contemplates use-limited products and the like to target hepatitis B virus infections. Accordingly, in a further aspect the present invention provides at least one immunogenic peptide or an immunogenic peptide group as described herein for use in treating hepatitis B virus infection in a human subject requiring treatment.
  • the present invention provides a vaccine composition as described herein for use in treating hepatitis B virus infection in a human subject requiring treatment.
  • the present invention provides an immunogenic peptide, immunogenic peptide group and/or a vaccine composition as described herein for the manufacture of a medicament for treating hepatitis B virus infection in a human subject requiring treatment.
  • the present invention provides the use of an immunogenic peptide, an immunogenic peptide group and/or a vaccine composition as described herein for treating a hepatitis B virus infection in a human subject requiring treatment.
  • the present invention further contemplates methods for generating peptide-loaded antigen presenting cells for use in (e.g.) autologous cell therapies.
  • the present invention provides a method for generating a peptide-loaded antigen-presenting cell, the method comprising contacting an antigen-presenting cell with:
  • the present invention further contemplates autologous cell therapies to treat hepatitis B infection.
  • the present invention provides a method of autologous cell therapy in a patient infected with a hepatitis B virus, the method comprising:
  • the present invention provides a pharmaceutical composition comprising a peptide-loaded antigen presenting cell, as described herein, for use in treating a hepatitis B infection in a patient.
  • the immunogenic peptide or peptide group is selected from any one of SEQ ID Nos: 1-11 and 111-142, and includes all combinations thereof.
  • the antigen-presenting cell is selected from a dendritic cell, a macrophage and a B cell.
  • the sample obtained from the patient is selected from plasma, serum and whole blood.
  • the peptide-loaded antigen presenting cell is cultured at between about 36 °C and about 45 °C, preferably between about 36 °C and about 38 °C, and most preferably at about 37 °C.
  • the peptide loaded antigen-presenting cell is cultured for between about 15 minutes and between about 5 hours.
  • the population of activated antigen-presenting cells comprises between about 10 5 activated antigen-presenting cells, about 10 6 activated antigen-presenting cells, about 10 7 activated antigen-presenting cells, about 10 8 activated antigen-presenting cells, about 10 9 activated antigen-presenting cells and about 10 10 activated antigen-presenting cells.
  • the activated antigen-presenting cells are administered to the patient by infusion.
  • an immunogenic peptide(s) or vaccine/composition(s) will depend upon the age, sex, health, and weight of the recipient, the kind of concurrent treatment, if any, the frequency of treatment, and the nature of the effect desired. However, the most preferred dosage can be tailored to the individual subject, as determined by the researcher or clinician.
  • the total dose required for any given treatment will commonly be determined with respect to a standard reference dose as set by a manufacturer, such as is commonly done with vaccines, such dose being administered either in a single treatment or in a series of doses, the success of which will depend on the production of a desired immunological result (i.e., successful production of a CTL-mediated response to the antigen, which response gives rise to the prevention and/or treatment desired).
  • a desired immunological result i.e., successful production of a CTL-mediated response to the antigen, which response gives rise to the prevention and/or treatment desired.
  • the overall administration schedule must be considered in determining the success of a course of treatment and not whether a single dose, given in isolation, would or would not produce the desired immunologically therapeutic result or effect.
  • Typical therapeutic doses lie within the range of from 1.0 pg to about 5,000 pg of peptide for a 70 kg patient, followed by boosting dosages of from about 1.0 pg to about 1,000 pg of peptide pursuant to a boosting regimen over days, weeks or even months, depending on the recipient's response and as necessitated by subsequent monitoring of CTL-mediated activity within the bloodstream.
  • dosages should be considered only a general guide and, in a given situation, may greatly exceed such suggested dosage regimens where the clinician believes that the patient's condition warrants more aggressive administration schedule.
  • the efficacy of administering additional doses, and of increasing or decreasing the interval may be re-evaluated on a continuing basis, in view of the recipient's immunocompetence.
  • the immunogenic vaccine compositions according to the present invention may be used against HBV infection by administration to a patient by a variety of routes.
  • the composition may be administered parenterally or orally, and, if parenterally, either systemically or topically.
  • Parenteral routes include subcutaneous, intravenous, intradermal, intramuscular, intraperitoneal, intranasal, transdermal, or buccal routes.
  • Other routes of parenteral administration include, for example, by bolus injection or by gradual perfusion over time.
  • vaccines are prepared as injectables, in the form of aqueous solutions or suspensions.
  • Vaccines in an oil base are also well known such as for inhaling.
  • Solid forms which are dissolved or suspended prior to use may also be formulated.
  • Pharmaceutical carriers, diluents and excipients are generally added that are compatible with the active ingredients and acceptable for pharmaceutical use. Examples of such carriers include, but are not limited to, water, saline solutions, dextrose, or glycerol. Combinations of carriers may also be used. These compositions may be sterilised by conventional, well known sterilisation techniques including sterile filtration.
  • Vaccine compositions may further incorporate additional substances to stabilize pH, or to function as adjuvants, wetting agents, or emulsifying agents, which can serve to improve the effectiveness of the vaccine.
  • the concentration of the T cell stimulatory peptides of the invention in pharmaceutical formulations are subject to wide variation, including anywhere from less than 0.01% by weight to as much as 50% or more. Factors such as volume and viscosity of the resulting composition must also be considered.
  • the solvents, or diluents, used for such compositions include water, possibly PBS (phosphate buffered saline), or saline itself, or other possible carriers or excipients.
  • the immunogens of the present invention may also be contained in artificially created structures such as liposomes, ISCOMS, slow-releasing nanoparticles, and other vehicles which increase the immunogenicity and/or half-life of the peptides or polypeptides in serum.
  • Liposomes include emulsions, foams, micelles, insoluble monolayers, liquid crystals, phospholipid dispersions, lamellar layers and the like.
  • Liposomes for use in the invention are formed from standard vesicle-forming lipids which generally include neutral and negatively charged phospholipids and a sterol, such as cholesterol. The selection of lipids is generally determined by considerations such as liposome size and stability in the blood.
  • Liposomes containing the peptides or polypeptides of the invention can be directed to the site of lymphoid cells where the liposomes then deliver the selected immunogens directly to antigen presenting cells.
  • Targeting can be achieved by incorporating additional molecules such as proteins or polysaccharides into the outer membranes of said structures, thus resulting in the delivery of the structures to particular areas of the body, or to particular cells within a given organ or tissue.
  • targeting molecules may be a molecule that binds to receptor on antigen presenting cells.
  • an antibody that binds to CD80 could be used to direct liposomes to dendritic cells.
  • the vaccines according to the present invention may also be administered as solid compositions.
  • Conventional nontoxic solid carriers including pharmaceutical grades of mannitol, lactose, starch, magnesium, cellulose, glucose, sucrose, sodium saccharin, and the like.
  • Such solid compositions will often be administered orally, whereby a pharmaceutically acceptable nontoxic composition is formed by incorporating the peptides and polypeptides of the invention with any of the carriers listed above.
  • such compositions will contain 10-95% active ingredient, and more preferably 25-75% active ingredient.
  • Aerosol administration is also an alternative, requiring only that the immunogens be properly dispersed within the aerosol propellant.
  • Typical percentages of the peptides or polypeptides of the invention are 0.01%-20% by weight, preferably l%-10%.
  • the use of a surfactant to properly disperse the immunogen may be required.
  • Representative surfactants include the esters or partial esters of fatty acids containing from 6 to 22 carbon atoms, such as caproic, octanoic, lauric, palmitic, stearic, linoleic, linolenic, olesteric and oleic acids with an aliphatic polyhydric alcohol or its cyclic anhydride.
  • esters such as mixed or natural glycerides may be employed.
  • the surfactant may constitute 0.1-20% by weight of the composition, preferably 0.25-5%.
  • Typical propellants for such administration may include esters and similar chemicals but are by no means limited to these.
  • a carrier, such as lecithin for intranasal delivery, may also be included.
  • a pharmaceutically acceptable salt or “pharmacologically acceptable salt” refers to a derivative of the disclosed immunogenic peptides wherein the peptide is modified by making acid or base salts of the agent.
  • acid salts are prepared from the free base (typically wherein the neutral form of the drug has a neutral — NH2 group) involving reaction with a suitable acid.
  • Suitable acids for preparing acid salts include both organic acids, e.g., acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p- toluenesulfonic acid, salicylic acid, and the like, as well as inorganic acids, e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid phosphoric acid and the like.
  • basic salts of acid moieties which may be present on a peptide are prepared using a pharmaceutically acceptable base such as sodium hydroxide, potassium hydroxide, ammonium hydroxide, calcium hydroxide, trimethylamine or the like.
  • the vaccine compositions of the present invention may include sugars, sugar alcohols, amino acids such as glycine, arginine, glutaminic acid and others as framework former.
  • the sugars may be mono-, di- or trisaccharide. These sugars may be used alone, as well as in combination with sugar alcohols. Examples of sugars include glucose, mannose, galactose, fructose or sorbose as monosaccharides, saccharose, lactose, maltose or trechalose as disaccharides and raffinose as a trisaccharide.
  • a sugar alcohol may be, for example, manniose
  • Preferred ingredients are saccharose, lactose, maltose, trehalose, mannitol and/or sorbitol, and more preferably, mannitol.
  • the vaccine compositions according to the present invention may include physiological well tolerated excipients (see Handbook of Pharmaceutical Excipients, 5. sup. th ed., edited by Raymond Rowe, Paul Shesky and Sien Owen, Pharmaceutical Press (2006), such as antioxidants like ascorbic acid or glutathione, preserving agents such as phenole, m- cresole,methyl-or propylparabene, chlorobutanol, thiomersal or benzalkoiumchloride, stabilizer, framework former such as saccharose, lactose, maltose, trehalose, mannitose, mannit and/or sorbit, mannit and/or lactose and solubilizer such as polyethyleneglycols (PEG), i.e.
  • physiological well tolerated excipients see Handbook of Pharmaceutical Excipients, 5. sup. th ed., edited by Raymond Rowe, Paul Shesky and Sien Owen, Pharmaceutical Press (2006), such as antioxidants like ascorbic acid
  • compositions of the present invention include one or more well tolerated excipients, selected from the group consisting of antioxidants, framework formers and stabilisers.
  • the pharmaceutical composition may also be formulated into a "unit dosage form", which used herein refers to physically discrete units, wherein each unit contains a predetermined quantity of a peptide or peptide combination, optionally in association with a pharmaceutical carrier (excipient, diluent, vehicle or filling agent) which, when administered in one or more doses, may produce a desired effect.
  • Unit dosage forms also include, for example, ampules and vials, which may include a composition in a freeze-dried or lyophilized state (a lyophilizate) or a sterile liquid carrier, for example that can be added prior to administration or delivery in vivo.
  • Unit dosage forms additionally include, for example, ampules and vials with liquid compositions disposed therein.
  • kits comprising a compartment and instructions, wherein the compartment comprises a pharmaceutical composition as described herein and wherein the instructions are for use in treating a hepatitis B virus infection.
  • a kit may further comprise packaging material comprising corrugated fiber, glass, plastic, foil, ampules, vials, blister pack, preloaded syringes or tubes, optionally that maintain sterility of the components.
  • a kit may further comprise labels or inserts comprising printed matter or computer readable medium optionally including identifying components, dose amounts, clinical pharmacology and instructions for the clinician or for a subject using one or more of the kit components, prophylactic or therapeutic benefits, adverse side effects or manufacturer information.
  • the kit additionally comprises a container comprising a solvent for dissolving the composition before use.
  • suitable solvents are described supra.
  • the kit may also comprise a device for use in parenteral injection, e.g. for injecting the composition (e.g. dissolved composition) to a subcutaneous or intradermal tissue.
  • a device may be any suitable device for that purpose, such as a needle or microneedle adapted for intradermal or subcutaneous delivery of the composition.
  • the device may be a microneedle or a device comprising a plurality of microneedles designed for intradermal delivery of liquids, e.g. as described in international patent applications WO14064543 Al, W005049107 A2, W006054280 A2, W007066341 A3 and WO14188429 Al.
  • the immunogenic peptides disclosed herein can be synthesised in solution or on a solid support in accordance with conventional techniques.
  • Various automated peptide synthesizers are commercially available and can be used in accordance with known protocols. See, for example, (Grant, G. A., Synthetic Peptides: A User's Guide, 1792, W. H. Freeman and Company, New York; Coligan, J. E. et al, Current Protocols in Protein Science, 1799, John Wiley & Sons, Inc., New York).
  • Fragments of immunogenic peptide according to the present invention may also be synthesised as intermediates in the synthesis of a larger polypeptide.
  • the immunogenic peptides and peptide variants are typically manufactured synthetically.
  • peptides, variants, derivatives and salts thereof are synthetic.
  • the peptides, variants, derivatives and salts may be isolated and/or purified, e.g. made by the hand of man, such as by peptide synthesis.
  • the peptides may be combined after synthesis and freeze-dried or dissolved in aqueous solutions, DMSO, glycerol or the like or mixtures thereof.
  • each peptide of a peptide combination is present in equimolar concentrations or substantially equimolar ranges.
  • the peptides are freeze-dried (lyophilized), such as to provide them in a storage-stable form and in a form ready to be re-dissolved.
  • concentration of each of the re-dissolved peptides may be in a molar concentration in the range of 1 to 1000 pM, for example in the range of 10 to 800 pM, for example in the range of 20 to 500 pM, for example in the range of 20 to 300 pM.
  • recombinant DNA technology may be employed wherein a nucleotide sequence which encodes an immunogenic peptide of interest is inserted into an expression vector, transformed or transfected into an appropriate host cell, and cultivated under conditions suitable for expression.
  • These procedures are well known to persons skilled in the art, (e.g.) as described in Coligan, J. E. et al, Current Protocols in Immunology, 1799, John Wiley & Sons, Inc., New York; Ausubel, F. M. et al, Current Protocols in Molecular Biology, 1799, John Wiley & Sons, Inc., New York; Molecular Cloning: A Laboratory Manual, 1789, Cold Spring Harbor Laboratory Press, Cold Spring Harbor.
  • recombinantly produced peptides may be used as immunogenic peptides according to the present invention.
  • the coding sequences for the immunogenic peptides of the length contemplated herein can be synthesised on commercially available automated DNA synthesisers using protocols that are well known in the art. See for example, Grant, G. A., Synthetic Peptides: A User's Guide, 1792, W. H. Freeman and Company, New York; Coligan, J. E. et al, Current Protocols in Protein Science, 1799, John Wiley & Sons, Inc., New York.
  • the coding sequences can also be modified such that an immunogenic peptide will be produced that incorporates a desired amino acid substitution, deletion or insertion.
  • the coding sequence can be provided with appropriate linkers, be ligated into suitable expression vectors that are commonly available in the art, and the resulting DNA or RNA molecule can be transformed or transfected into suitable hosts to produce the desired fusion protein.
  • suitable host systems are available, and their selection is left to the skilled person.
  • the coding sequence will be provided with operably linked start and stop codons, promoter and terminator regions, and a replication system to provide an expression vector for expression in the desired host cell.
  • promoter sequences compatible with bacterial hosts are provided in plasmids containing convenient restriction sites for insertion of the desired coding sequence.
  • the resulting expression vectors are transformed into suitable bacterial hosts, although yeast, insect, and mammalian host cells may also be used, employing suitable vectors and control sequences.
  • Host cells are genetically engineered (transduced or transformed or transfected) with the vectors of this invention which may be, for example, a cloning vector or an expression vector.
  • the vector may be, for example, in the form of a plasmid, a viral particle, a phage, etc.
  • the engineered host cells can be cultured in conventional nutrient media modified as appropriate for activating promoters, selecting transformants or amplifying the genes of the present invention.
  • the culture conditions such as temperature, pH and the like, are those previously used with the host cell selected for expression, and will be apparent to the skilled person.
  • the present invention also includes recombinant constructs comprising one or more of the sequences as broadly described above.
  • the constructs comprise a vector, such as a plasmid or viral vector, into which a sequence of the invention has been inserted, in a forward or reverse orientation.
  • the construct further comprises regulatory sequences, including, for example, a promoter, operably linked to the sequence.
  • a promoter operably linked to the sequence.
  • the present invention further relates to host cells containing the above-described constructs.
  • the host cell can be a higher eukaryotic cell, such as a mammalian cell, or a lower eukaryotic cell, such as a yeast cell, or the host cell can be a prokaryotic cell, such as a bacterial cell.
  • Introduction of the construct into the host cell can be effected by calcium phosphate transfection, DEAE-Dextran mediated transfection, or electroporation (Ausubel, F. M. et al, Current Protocols in Molecular Biology, 1799, John Wiley & Sons, Inc., New York; Molecular Cloning: A Laboratory Manual, 1789, Cold Spring Harbor Laboratory Press, Cold Spring Harbor).
  • Such cells can routinely be utilised for assaying CTL activity by having said genetically engineered, or recombinant, host cells express the immunogenic peptides of the present invention.
  • mammalian cell culture systems can also be employed to express recombinant protein.
  • mammalian expression systems include the COS-7 lines of monkey kidney fibroblasts, described by Gluzman, Cell, 23: 175 (1781), and other cell lines capable of expressing a compatible vector, for example, the C127, 3T3, CHO, HeLa and BHK cell lines.
  • Mammalian expression vectors will comprise an origin of replication, a suitable promoter and enhancer, and also any necessary ribosome binding sites, polyadenylation site, splice donor and acceptor sites, transcriptional termination sequences, and 5' flanking nontranscribed sequences. DNA sequences derived from the SV40 splice, and polyadenylation sites may be used to provide the required nontranscribed genetic elements.
  • the polypeptide can be recovered and purified from recombinant cell cultures by methods including ammonium sulfate or ethanol precipitation, acid extraction, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxylapatite chromatography and lectin chromatography. Protein refolding steps can be used, as necessary, in completing configuration of the mature protein. Finally, high performance liquid chromatography (HPLC) can be employed for final purification steps.
  • HPLC high performance liquid chromatography
  • the IEDB (www.iedb.org) was queried in late August 2018 for linear peptide sequences derived from Hepatitis B virus (organism ID 10407) that have been reported in the published literature as having the capacity to elicit an immune response restricted by human (HLA) class I MHC. Accordingly, the query specified humans (taxonomy organism ID 9606) as the host organism. HLA transgenic mice were also included as an alternative source organism because a significant amount of published data relates to immunogenicity of HBV epitopes in HLA transgenic mice. Further, the query was limited to T cell epitopes associated with positive responses and associated with class I MHC restriction. This query returned 177 unique epitopes, associated with 901 different assays, reported in 118 references.
  • the "Effector cells” column highlights whether the T cells used to detect epitope responses were assayed directly ex vivo, or after a short term in vitro restimulation, or were long term lines/clones. The rationale for this distinction is that ex vivo determinations are deemed most representative of physiological conditions, followed by short term in vitro restimulation, while data obtained with cloned cells are the least representative.
  • the "Assay type” column refers to our arbitrary subjective preference in terms of assays used to detect epitope responses.
  • This characterization considers, in order of decreasing interest, multimer/tetramer staining as preferred over readouts based on intracellular cytokine staining (ICS), enzyme linked immune absorbent spot detection (ELISPOT) or 51Cr-release; less weight was given to proliferation or ELISA based assays (see Carrasco Pro et al., PMID 763461). Accordingly, the "Assay type” column indicates the “best available assay” associated with each epitope.
  • ICS cytokine staining
  • ELISPOT enzyme linked immune absorbent spot detection
  • 51Cr-release 51Cr-release
  • HBV-derived HLA class I restricted T cell epitopes were examined to identify particular clusters of epitopes encompassing stretches of approximately 32 contiguous amino acids.
  • Each epitope was aligned against a corresponding HBV reference protein sequence.
  • the reference sequences were specific for the external core protein (P0C767, inclusive of the capsid protein), large envelope protein (Q76R62), protein P (Q69028) and protein X (Q69027), and represent the HBV reference sequence set utilized by the IEDB. The starting positions of all epitopes were indexed to the respective reference sequence, and then the epitopes sorted accordingly.
  • clusters In total, 35 different clusters were identified. The clusters ranged in inclusiveness from just one epitope to as many six and ranged in length from nine residues up to 32. Exemplary clusters were shown in Table 1, sorted in descending order on the basis of estimated population coverage. The protein of provenance and respective start and end positions in the corresponding reference sequence was also shown. The table also indicated the length of each cluster and the number of associated epitopes. Further, the total number of different positive assay responses associated with the epitopes in the cluster, and the number of unique restricting HLA alleles, was shown.
  • Clusters were found to be associated with population coverage >50%.
  • One of these clusters is from Protein X, which has not been associated with robust immunological responses, even in acutely infected patients.
  • Another cluster consists of a single epitope from Protein P which did not map to the Protein P reference sequence. Accordingly, these two clusters were removed from further consideration. The remaining nine clusters includes all but one cluster with four or more unique HLA class I restrictions, and all clusters with more than 30 total responses.
  • Four of the clusters are derived from the capsid/external core protein, three of the clusters are derived from the envelope protein and two clusters are derived from protein P.
  • the expected population coverage for the entire set of 86 epitopes is 90.2%.
  • the "Top 8 clusters" in terms of population coverage at the time of analysis i.e. Cluster 3 (SEQ ID NO: 2), Cluster 25 (SEQ ID NO: 3), Cluster 28 (SEQ ID NO: 4), Cluster 8 (SEQ ID NO: 5), Cluster 13 (SEQ ID NO: 7), Cluster 2 (SEQ ID NO: 8), Cluster 5 (SEQ ID NO: 9) and Cluster IB (SEQ ID NO: 10)
  • the "Top 6 clusters" at the time of analysis i.e.
  • Cluster 3 (SEQ ID NO: 2), Cluster 25 (SEQ ID NO: 3), Cluster 28 (SEQ ID NO: 4), Cluster 8 (SEQ ID NO: 5), Cluster 5 (SEQ ID NO: 9) and Cluster IB (SEQ ID NO: 10)) while having slightly fewer responses achieved a comparable level of population coverage with 88.5%.
  • both 'sets' of clusters achieved a very similar level of population coverage (i.e.) 89.8% and 88.5% versus 90.2%.
  • the level of population coverage i.e. by geographical area or by ethnicity is expected to improve if similar calculations were run using subsequently developed clusters, namely Cluster 1 and Cluster 8B.
  • Applicants then determined the minimal expected population coverage for representative vaccine constructs designed following the cluster approach described in Example 2.
  • the population coverage tool hosted by the IEDB (Bui et al., PMID 16545123) was to calculate the average population coverage worldwide, and in Caucasian (i.e. European) ethnicities.
  • Each cluster construct was assessed individually for coverage in the general worldwide population, and also the Caucasian population.
  • the results are shown in Table 6, below; the results are tabulated including only non-redundant epitopes for each cluster that comport with a consensus sequence for the corresponding cluster, and thus may represent conservative estimates). More specifically, as variant epitopes were not considered, and only epitopes with exact sequence matches were included, the number of epitopes evaluated for some clusters are lower than shown in Tables 1 and 2.
  • Example 1 The information presented in Example 1 identified a set of 86 HBV derived HLA class I restricted T cell epitopes that have been described in the published literature. Further analysis of the data identified a subset of 24 epitopes that were positive in at least 4 assays and in at least 2 independent reports, and that were also associated with responses detected ex vivo. These epitopes were proposed as lead candidates for inclusion in vaccine bead construct design.
  • each epitope bead would contain at least one HLA-A*02:01 restricted epitope.
  • SUBSTITUTE SHEET (RULE 26) In terms of construct design, Applicants first separated the top eight (8) A*02:01 epitopes, defined on the basis of the number of responses, from the "remainders", utilizing each as the anchor of respective beads. In constructing the beads, Applicants considered two broad approaches, with the first prioritizing inclusion of the best epitopes in the same bead (i.e. SEQ ID Nos: 111-118 and 127-134), and the second attempting to develop a more balanced set of beads, including in each bead some of the more dominant and less dominant epitopes in terms of total responses (i.e. SEQ ID Nos: 119-125 and 135-142).
  • Applicants also considered two variations for combing the individual epitopes, with the first linking consecutive epitopes using double alanine spacers (i.e. SEQ ID Nos: 111- 126), and the second linking epitopes using the respective natural (native) N- and C-terminal flanking residue of each epitope (i.e. SEQ ID Nos: 127-142).
  • constructs 17-24 are shown as constructs 17-24, respectively, in Table 4C.
  • bead constructs 1-16 in Tables 4A and 4B similarly pertain to the corresponding beads construct 17-32 in Tables 4C and 4D; that is, the individual epitope sequences, total responses, restricting HLA alleles, and total population coverage are the same-only the spacers are different.
  • the best bead constructs namely bead 1 (SEQ ID NO: 111) and bead 17 (SEQ ID NO: 127) was comprised of three epitopes from the capsid protein. This bead tallied 191 responses, accounting for 25.1% of the total reported HBV response. Twelve different HLA alleles were associated with these responses, and the string provides population coverage of almost 74%.
  • the next best bead constructs namely bead 2 (SEQ ID NO: 112) and bead 18 (SEQ ID NO: 128), accounted for 12% of the response, with population coverage of about 47%.
  • the remaining bead constructs provided from 36 to 62% coverage, and accounted for between 2.1 and 11.1% of the response.
  • the second approach to generating a set of beads aimed to assemble a more balanced panel, where the fraction of the total HBV response would be more evenly distributed across the various beads.
  • the Applicant matched the best A*02:01 epitope with the "worst" (in terms of total responses) of the remainders (i.e. SEQ ID Nos: 119 and 135).
  • a second bead was built with the second best A*02:01 epitope matched with the second "worst" of the remainders (i.e. SEQ ID Nos: 120 and 136) etc.
  • Table 4E Population Coverage for Discrete Beads/Bead Combinations
  • the B07-supertype epitope has been treated as a separate epitope in constructing the various beads (refer to bead construct 2 (SEQ ID NO: 112), bead construct 15 (SEQ ID NO: 125), bead construct 18 (SEQ ID NO: 128) and bead construct 31 (SEQ ID NO: 141)), but its coverage has also been enhanced when the longer peptide is considered (refer to bead construct 1 (SEQ ID NO: 111), bead construct 9 (SEQ ID NO: 119), bead construct 17 (SEQ ID NO: 127) and bead construct 25 (SEQ ID NO: 135).
  • SUBSTITUTE SHEET (RULE 26) string of two with the most responses (i.e. SEQ ID Nos: 121 and 137), and so on.
  • SEQ ID Nos: 121 and 137 SEQ ID Nos: 121 and 137
  • bead constructs 9-16 in Table 4B and bead constructs 25-32 in Table 4D were assembled. These eight constructs (16 total taking account of the different spacers used) ranged for 4.9 to 22.4% of the total HBV response, and provided between 39.6 and 62.5% population coverage. Achieving more balance in the set is limited by the dominance of the HBV capsid epitope (FLPSDFFPSV; SEQ ID NO: 12).
  • Table 4E Population Coverage for Discrete Beads/Bead Combinations
  • the B07-supertype epitope has been treated as a separate epitope in constructing the various beads (refer to bead construct 2 (SEQ ID NO: 112), bead construct 15 (SEQ ID NO: 125), bead construct 18 (SEQ ID NO: 128) and bead construct 31 (SEQ ID NO: 141)), but its coverage has also been enhanced when the longer peptide is considered (refer to bead construct 1 (SEQ ID NO: 111), bead construct 9 (SEQ ID NO: 119), bead construct 17 (SEQ ID NO: 127) and bead construct 25 (SEQ ID NO: 135).
  • the average HLA coverage of the eight different constructs was 49.7% ( ⁇ 13.0%), and these constructs covered 8.7% ( ⁇ 7.5%) of the total response.
  • the HLA coverage was 48.7% ( ⁇ 9.4%), and 8.7% ( ⁇ 5.9%) of the total response. Note that in comparing the % of response covered by the "more dominant" approach with the "balanced” approach, the average is the same by definition, since the same number of responses are being spread across the same number of constructs. The % population coverage could, however, vary and indeed does, if only by a small amount.
  • cluster approach provides HLA and response coverage similar to that provided by the bead constructs derived from the isolated epitopes.
  • HBV helper T-cell lymphocyte
  • IEDB IEDB for HBV-derived HLA class II restricted epitopes.
  • HTL epitopes were identified for all of the cluster constructs, with the exceptions of Clusters 25 and 28. Refer to Table 7, below. Notably, Clusters 5 and 13 contained 3 epitopes each. These epitopes would be expected to provide HBV-specific help to the respective constructs. Thus, taken together, half of the set of constructs are associated with some known HBV-derived HTL epitopes. Table 7: Known HBV HTL Epitopes with Identity to Construct Sequences
  • the various CD4+ T-helper cell epitopes associated with each cluster are identified in Table 7A, on the following page.
  • each construct was parsed into constituent 15-mers and assessed for predicted MHC class II binding capacity for a panel of 27 common and representative HLA DR, DQ and DP molecules (Greenbaum et al., PMID 21305276). Peptides scoring in the top 10th percentile using the IEDB consensus algorithm were considered binders for the respective class II specificity.
  • binding predictions suggest that each cluster construct should be associated with more than about 10 different possible peptide/MHC binding events, or about 80 on average, which have the potential to provide construct specific help.
  • Cluster 8B Identification of a Population Specific Cluster
  • This example describes the analyses necessary for evaluating potential benefit of designing an additional HBV cluster construct to include the HBV envelope 182-191 epitope, highlighted in a paper by Tan et al. (PMID 24227846) as an immunodominant HLA-C*08:01 restricted T cell epitope in the context of a cohort of HBV infected subjects in Southeast Asia.
  • the HBV envelope 182-191 epitope (SEQ ID NO: 36) does not align with any of the other cluster constructs presently under consideration as vaccine constructs. However, it is just 12 residues to the N-terminal side of Cluster 8. Accordingly, Applicants examined the potential coverage of a new cluster, Cluster 8B, that represents a frame shift of Cluster 8. By shifting the frame of Cluster 8 (envelope 194-223) to now encompass residues 182-215 (Table 8), including the C*08:01 epitope (182-191), one T-cell epitope (215-223) recognised by A*02:01 and A*24:02 is lost. However, the loss of coverage provided by 215-223 is mitigated by the fact that other epitopes included in both Cluster 8 and 8B cover the same alleles.
  • this frameshift also incorporates an additional A*02:01 epitope in the region of residues 188-196.
  • Cluster 8B contains 7 different HLA/epitope combinations, compared to 6 for Cluster 8. Further, the total number of associated positive responses reported in the literature is 95 for the new cluster 8B, versus 81 for Cluster 8. Thus, Cluster 8B potentially provides greater coverage than Cluster 8.
  • Cluster 8B summarises characteristics of Cluster 8B, including population coverage, in comparison with the previously described clusters.
  • Cluster 8B is the longest sequence, covering 35 residues. Its potential population coverage, both worldwide and in Caucasians, is around 60%, similar to coverage provided by Cluster 8, and lower than afforded by several other clusters (1, 3, 25 and 28) which provide coverage in the 68-80% range, depending on population.
  • Interest in the HLA C*08:01 restricted envelope 182-191 epitope was prompted on the basis of its high response rate in a Southeast Asian cohort, potentially relevant for vaccine trials.
  • Cluster 8B is predicted to provide relatively high coverage (68%) in the general Southeast Asian population (Borneo, Philippines, Indonesia, Singapore, Malaysia, Vietnam, Thailand, Taiwan); only Clusters 3 (83%) and 28 (76%) are predicted to provide higher coverage in the same population.
  • Cluster 8B retains the same two dominant HTL epitopes associated with Cluster 8, but also incorporates a 17-mer overlap with a promiscuous HTL epitope spanning residues 180-198 of the envelope sequence.
  • Table 11 provides a comparison between all of the candidate clusters in terms of HTL capacity.
  • Cluster 8B is associated with more defined HLA restrictions (10) and predicted HLA class II binding events (230) than all of the other HBV clusters.
  • PBMC peripheral blood mononuclear cell
  • PBMCs Plasma samples were obtained by New Zealand Clinical Research from chronic HBV patients (27), acute HBV patients (3) and healthy subjects (15). PBMCs were purified from the blood samples and cryopreserved.
  • PBMCs were then thawed and tested for T cell responses to the 10 peptides.
  • Methodology was fluorescent ELISpot ("fluorospot” or “FSPOT” below) to quantify cells secreting interferon-gamma (IFN-y) in response to each peptide, following a pre-stimulation and 7-day culture period to enable expansion of antigen-specific T cells.
  • Cluster 13 and Cluster 8b Two candidate peptides (Cluster 13 and Cluster 8b) induced responses primarily in HBV patients.
  • Two candidate peptides (Bead 17 and Cluster 1) induced responses in a higher proportion of HBV patients, and of a greater magnitude, than in healthy controls.
  • Cluster 3 and Cluster 28 Two candidate peptides (Cluster 3 and Cluster 28) induced responses in similar proportions of HBV patients and healthy controls, although the magnitude of the responses was higher in HBV patients.
  • Cluster 2 and Cluster 25 Two candidate peptides (Cluster 2 and Cluster 25) induced responses in similar proportions of HBV patients and healthy controls, of similar magnitudes.
  • Cluster 8B SEQ ID NO: 6
  • Cluster 13 SEQ ID NO: 7
  • the present example summarizes analyses to update and expand estimates of potential HLA class I population coverage of the three lead candidates, as well as the various envelope candidates, original and novel (Tables 13 and 14). Besides the general worldwide population, and the Caucasian population representative of the majority of the preceding epitope identification studies, the present analyses examine coverage in China and other Asian populations, but also the Americas and Africa.
  • the initial panel of constructs considered as candidates for vaccine development included two, Cluster 8b (pep447) and Cluster 13 extended (pep449), based on HLA class I restricted T cell epitopes derived from the envelope protein. Subsequent analyses of these constructs in HLA-A2 transgenic mice and human PBMC studies, and with consideration towards manufacturability, suggested a need to design and develop additional envelopebased constructs. Accordingly, Applicants examined the sequences of the two constructs to identify residues that could be eliminated to allow shortening the construct, without elimination of significant allele coverage.
  • pep447 we identified three possible avenues.
  • pep447 R1 see Table 12
  • Applicants eliminated the first 11 residues at the N-terminus. Functionally, this eliminated two less dominant, but redundant, A*02:01 restricted epitopes.
  • An HLA C*08:01 epitope was also eliminated with this deletion. However, as HLA-C responses are believed to be likely less crucial than A- or B-restricted ones, this loss may be mitigated by increased manufacturability.
  • pep447 R2 elimination of the first 6 N-terminal residues may be considered. This deletion removes one A*02:01 and C*08:01 epitope, but retains a second A*02:01 epitope.
  • pep447 R2 could consider elimination of the naturally occurring residues PQSLD in the middle of the sequence. Elimination of this span would shorten the sequence by an additional five residues without impacting any of the known T cell epitopes contained in the construct, and thus would not impact population coverage.
  • pep449 Applicants considered elimination of the first 4 residues at the N-terminus, along with the last 2 residues at the C-terminus. These deletions, shortening the construct to 22 residues, eliminate less dominant but redundant A*02:01 and A*24:02 epitopes, while retaining the most widely reported epitopes for each allele. As a result, there is no change in population coverage (see below).
  • An additional alternative (not shown) that could be considered, and that would also not impact population coverage, would be elimination of only the first two residues at the N-terminus and the middle 4 residues QWFV. These deletions would similarly reduce the sequence to 22 residues, but retain additional dominant A*02:01 and A*24:02 epitopes.
  • E new bead 1 Applicants combined a dominant A*02:01 epitope that largely overlaps with one of the most reported A*24:02 epitopes, with another dominant A*02:01 epitope.
  • HLA A*ll :01 epitope that has been reported in only a single study.
  • this epitope was characterized using HLA A*ll :01 tetramers, and had been assayed, and gave good responses, with chronic hepatitis B (CHB) patients in the Chongqing area of China.
  • CHB chronic hepatitis B
  • a second alternative novel bead "E new bead 2" combined two dominant A*02:01 epitopes, one of which is also restricted by HLA C*08:01, with an A*24:02 epitope.
  • E new bead 2 combined two dominant A*02:01 epitopes, one of which is also restricted by HLA C*08:01, with an A*24:02 epitope.
  • Applicants predicted the potential population coverage afforded by various constructs in the general worldwide and Caucasian populations. Here, Applicants have expanded the coverage analysis to include several different population groups. At the same time, Applicants have evaluated the potential coverage predicted for the novel envelope epitope-based constructs described above.
  • the population coverage tool hosted by the IEDB (Bui et al., PMID 16545123) to calculate population coverage.
  • This tool calculates the fraction of individuals predicted to respond to a given epitope set on the basis of HLA genotypic frequencies associated with different populations, as derived from the Allele Frequency Net Database (AFND; www.allelefrequencies.net).
  • the tool computes the projected population coverage, defined as the number of individuals expected to recognize at least one or more of the epitopes in the set considered.
  • Applicants similarly considered the average coverage worldwide, and in Caucasians (i.e., Europeans), but also included several regions of Asia, to include Northeast Asia (China), and also Africa and the Americas.
  • E new bead 1 and 3 The bead approach to generation of envelope constructs did result in somewhat better coverage in the cases of E new bead 1 and 3.
  • increases of about 10% or more were observed in most populations.
  • coverage of 70% or better is predicted for both of these novel constructs for China, and generally across most of the other Asian populations considered.
  • E new bead 2 also afforded minor increases in some contexts, but in generally was little different than the two original constructs.

Abstract

La présente invention concerne de nouveaux vaccins thérapeutiques pour une maladie infectieuse. La présente invention concerne des peptides immunogènes, des groupes peptidiques immunogènes, des compositions de vaccin et des méthodes thérapeutiques ciblant une infection chronique par le virus de l'hépatite B (HBV) chez l'homme. Les peptides immunogènes comprennent un ensemble unique de sélection/combinatoire d'épitopes de lymphocytes T restreints par un antigène leucocytaire humain de classe I (HLA) pour produire des compositions de vaccin qui réalisent une couverture de population étendue, par exemple, par géographie et/ou ethnicité. Les compositions de vaccin selon la présente invention fourniront une solution bien requise pour la gestion de cette maladie chronique pour laquelle il n'existe actuellement aucun traitement curatif authentique.
PCT/NZ2022/050141 2021-11-09 2022-11-09 Nouveaux vaccins thérapeutiques WO2023085956A1 (fr)

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Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012109404A1 (fr) * 2011-02-12 2012-08-16 Globeimmune, Inc. Thérapeutique à base de levure pour infection chronique par l'hépatite b
WO2014102540A1 (fr) * 2012-12-24 2014-07-03 Immune Targeting Systems (Its) Limited Produit thérapeutique contre vhb
WO2015082905A1 (fr) * 2013-12-02 2015-06-11 Immune Targeting Systems (Its) Ltd Composé immunogène
WO2017121791A1 (fr) * 2016-01-12 2017-07-20 Helmholtz Zentrum München - Deutsches Forschungszentrum für Gesundheit und Umwelt (GmbH) Moyens et méthodes pour le traitement du vhb
WO2021045969A1 (fr) * 2019-08-29 2021-03-11 Vir Biotechnology, Inc. Vaccins contre le virus de l'hépatite b
WO2021067181A1 (fr) * 2019-09-30 2021-04-08 Gilead Sciences, Inc. Vaccins contre le virus de l'hépatite b et méthodes de traitement du vhb

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012109404A1 (fr) * 2011-02-12 2012-08-16 Globeimmune, Inc. Thérapeutique à base de levure pour infection chronique par l'hépatite b
WO2014102540A1 (fr) * 2012-12-24 2014-07-03 Immune Targeting Systems (Its) Limited Produit thérapeutique contre vhb
WO2015082905A1 (fr) * 2013-12-02 2015-06-11 Immune Targeting Systems (Its) Ltd Composé immunogène
WO2017121791A1 (fr) * 2016-01-12 2017-07-20 Helmholtz Zentrum München - Deutsches Forschungszentrum für Gesundheit und Umwelt (GmbH) Moyens et méthodes pour le traitement du vhb
WO2021045969A1 (fr) * 2019-08-29 2021-03-11 Vir Biotechnology, Inc. Vaccins contre le virus de l'hépatite b
WO2021067181A1 (fr) * 2019-09-30 2021-04-08 Gilead Sciences, Inc. Vaccins contre le virus de l'hépatite b et méthodes de traitement du vhb

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