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JP-7855515-B2 - Compositions and methods for treating and preventing hepatitis B and hepatitis D

JP7855515B2JP 7855515 B2JP7855515 B2JP 7855515B2JP-7855515-B2

Inventors

  • セルベルグ，マッティ
  • フレリン，ラーシュ

Assignees

  • スヴェンスカ ヴァクチンファブリケン プロダクション アーベー

Dates

Publication Date
20260508
Application Date
20210126
Priority Date
20200128

Claims (20)

  1. (a) a nucleic acid comprising at least one nucleic acid sequence encoding hepatitis D antigen (HDAg) and at least one nucleic acid sequence encoding PreS1; and (b) an immunogenic combination product comprising at least one HDAg polypeptide sequence and at least one PreS1 polypeptide sequence, An immunogenic combination product administered to a subject as an initial prime dose containing (a) nucleic acid, followed by one or more boost doses containing (b) polypeptide .
  2. The combination product according to claim 1, wherein at least one nucleic acid sequence encoding HDAg includes the sequence shown in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4, or any combination thereof.
  3. The combination product according to claim 1 or 2, wherein at least one nucleic acid sequence encoding PreS1 includes the sequence shown in SEQ ID NO: 9 or SEQ ID NO: 10, or both.
  4. The combination product according to any one of claims 1 to 3, wherein in the nucleic acid, each HDAg nucleic acid sequence is grouped with a PreS1 nucleic acid sequence, and in each group, the PreS1 nucleic acid sequence is located immediately downstream of the HDAg nucleic acid sequence .
  5. The combination product according to claim 4, further comprising at least one nucleic acid sequence encoding a self-catalytic peptide cleavage site , wherein the HDAg nucleic acid sequence and the PreS1 nucleic acid sequence are separated by at least one nucleic acid sequence encoding the self-catalytic peptide cleavage site.
  6. The combination product according to claim 5, wherein at least one nucleic acid sequence encoding the autocatalytic peptide cleavage site includes a nucleic acid sequence selected from the group consisting of the nucleic acid sequence of 2A (P2A) derived from porcine rhinitis virus type 1, the nucleic acid sequence of 2A (F2A) derived from foot-and-mouth disease virus, the nucleic acid sequence of 2A (E2A) derived from equine rhinitis A virus (ERAV), and the nucleic acid sequence of 2A (T2A) derived from Thosea asigna virus, and the encoded autocatalytic peptide cleavage site may include a GSG (glycine-serine-glycine) motif at its N-terminus.
  7. The combination product according to claim 5 or 6, wherein at least one nucleic acid sequence encoding the autocatalytic peptide cleavage site includes the sequence shown in Sequence ID No. 13.
  8. The combination product according to any one of claims 1 to 7, wherein the nucleic acid is codon-optimized for human expression.
  9. The combination product according to any one of claims 1 to 8, wherein the nucleic acid comprises a sequence having at least 95% sequence identity with any one of the sequences indicated by sequence numbers 15 to 24, 35, or 36.
  10. The combination product according to claim 9, wherein the nucleic acid includes a sequence having at least 95% sequence identity with any one of the sequences indicated by SEQ ID NO: 18, SEQ ID NO: 35, or SEQ ID NO: 36.
  11. The combination product according to any one of claims 1 to 10, wherein the at least one HDAg polypeptide sequence includes one of the sequences shown in SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, or SEQ ID NO: 8, or any combination thereof.
  12. The combination product according to any one of claims 1 to 11, wherein the at least one PreS1 polypeptide sequence includes the sequence shown in SEQ ID NO: 11 or SEQ ID NO: 12, or both.
  13. The combination product according to any one of claims 1 to 12, wherein the at least one PreS1 polypeptide sequence is located downstream of the at least one HDAg polypeptide sequence.
  14. The combination product according to any one of claims 1 to 13, wherein the polypeptide comprises a sequence having at least 95% sequence identity with any one of the sequences indicated by sequence numbers 25 to 34 or 37.
  15. The combination product according to any one of claims 1 to 14, wherein the polypeptide comprises a sequence having at least 95% sequence identity with any one of the sequences indicated by sequence numbers 29, 31, 32, or 37.
  16. The combination product according to any one of claims 1 to 15, wherein the polypeptide is recombinantly expressed.
  17. The combination product according to claim 16, wherein the polypeptide is recombinantly expressed in a mammalian, bacterial, yeast, insect, or cell-free system.
  18. The combination product according to any one of claims 1 to 17, further comprising an adjuvant.
  19. The combination product according to any one of claims 1 to 18, wherein the nucleic acid includes DNA.
  20. The combination product according to any one of claims 1 to 19, wherein the nucleic acid is provided in the form of a recombinant vector.

Description

This application, which cross-references related applications , claims the benefit of priority from U.S. Provisional Patent Application No. 62/966970, filed on 28 January 2020, which is expressly incorporated herein by reference in its entirety. This application, which references a sequence listing , was filed together with an electronic sequence listing. This sequence listing was created with the filename SVF005SeqListing.TXT, last modified on January 26, 2021, and has a file size of 141,377 bytes. The information contained in this electronic sequence listing is expressly incorporated herein by reference in its entirety. Aspects of the present disclosure, as a whole, relate to immunogenic compositions or immunogenic combination products comprising genetically modified HBV and HDV nucleic acids, genes, peptides or proteins, used to induce an immune response to hepatitis B virus (HBV) infection and/or hepatitis D virus (HDV) infection. This immune response includes, is essentially derived from, or comprises activated immune cells that produce neutralizing antibodies against HBV and/or HDV, as well as activated immune cells against HBV and/or HDV, such as T cells and B cells. The disclosure also relates, as a whole, to a method of using the above-mentioned immunogenic compositions or immunogenic combination products in a subject to induce an immune response to HBV and/or HDV, comprising administering the compositions or combination products in a homogeneous or heterogeneous inoculation method comprising a priming with nucleic acids and/or polypeptides and a boosting with nucleic acids and/or polypeptides. Hepatitis is a disease characterized by swelling and inflammation of the liver. This disease is generally caused by viruses, and currently five types (A, B, C, D, and E) are known. Hepatitis B infection can be acute or chronic, and severe chronic infection can lead to chronic inflammation, fibrosis, cirrhosis, and hepatocellular carcinoma. The hepatitis B virus (HBV) has an incomplete double-stranded circular DNA genome. Once translocated to the host nucleus, the genome is transcribed into four viral mRNA molecules by the host's RNA polymerase. These mRNA molecules are then translated into viral proteins such as capsid proteins and surface antigens, or reverse transcriptase generates a large DNA genome. Hepatitis D virus (HDV) is a virus that replicates by co-infection or co-infection with HBV. The circular single-stranded RNA of HDV is amplified by the host's RNA polymerase, but this RNA contains only one hepatitis D antigen (HDAg) gene. In co-infection with HBV and HDV, naked HDV is packaged in an envelope containing the HBV surface antigen, and the RNA genome coated with the HDAg protein is surrounded by this envelope. Since HDV does not encode its own receptor-binding protein, the uptake of the HBV surface antigen is essential for HDV infection. Co-infection with hepatitis D can lead to even more serious complications, increasing the risk of liver failure, cirrhosis, and liver cancer. There is currently a need for effective immunogenic compositions and vaccines to acquire immunity that prevents infection with both hepatitis B and hepatitis D. Further features and variations beyond those described above can be easily understood from the following drawings and exemplary embodiments. The following drawings illustrate representative embodiments and do not limit the scope of the present invention. These are nucleic acid constructs or polypeptide constructs containing HBV antigen and/or HDV antigen used herein. Ten constructs are presented: Delta-1 (Δ-1, D1), Delta-2 (Δ-2, D2), Delta-3 (Δ-3, D3), Delta-4 (Δ-4, D4), Delta-5 (Δ-5, D5), Delta-6 (Δ-6, D6), Delta-7 (Δ-7, D7), Delta-8 (Δ-8, D8), Delta-9 (Δ-9, D9), and Delta-10 (Δ-10, D10) (Figure 1A). Western blotting confirmed the proper expression of these ten polypeptide constructs (Figure 1B). GFP was used as a control for Western blotting. This is a construct used in the DNA prime/protein boost composition inoculation method. The DNA composition contains a Δ-4 nucleic acid sequence, and the protein composition contains a Δ-7 or Δ-8 polypeptide sequence, or a polypeptide sequence fused with Δ-7 and Δ-8. This study shows the results of quantifying interferon-gamma (IFNγ) formation spots per 10⁶ cells, which are indicators of T lymphocyte activation, by performing an ELISpot assay on a population of leukocytes purified from the serum of mice immunized with an HBV/HDV DNA composition, after exposure to various HBV or HDV antigens. The antigens used were PreS1 A (SEQ ID NO: 11), PreS1 A (SEQ ID NO: 12), HDAg genotype 1 A (SEQ ID NO: 5, "HDAg gtp 1A-pool 1" and "HDAg gtp 1A-pool 2"), HDAg genotype 1 B (SEQ ID NO: 6, "HDAg gtp 1B-pool 3" and "HDAg gtp 1B-pool 4"), HDAg genotype 2 A (SEQ ID NO: 7, "HDAg gtp 2C-pool 5" and "HDAg gtp 2C-pool 6"), or HDAg genotype 2 B (SEQ ID NO: 8, "HDAg gtp 2D-pool 7" and "HDAg gtp 2D-pool 6") This is a purified polypeptide containing "