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JP-2026514290-A - Nucleic acids and their use

JP2026514290AJP 2026514290 AJP2026514290 AJP 2026514290AJP-2026514290-A

Abstract

This specification discloses a self-amplifying RNA (saRNA) molecule encoding an influenza virus antigen and a method of using the same. [Figure 1]

Inventors

  • ピラダ スファヒハット アレン
  • イェ チェ
  • ラケル ムニョス－モレノ
  • アリシア ソロルザノ キハノ

Assignees

  • ファイザー・インク

Dates

Publication Date
20260508
Application Date
20240206
Priority Date
20230209

Claims (19)

  1. A composition comprising a first RNA molecule encoding a target or desired gene derived from an influenza virus, and a second RNA molecule encoding an influenza non-structural (NS1) protein, wherein the first polynucleotide is a self-amplifying RNA.
  2. The composition according to claim 1, wherein the second RNA molecule comprises a modified nucleotide.
  3. The composition according to any one of claims 1 to 2, wherein the second RNA molecule does not contain a subgenome promoter derived from an alphavirus.
  4. The composition according to claim 3, wherein the self-amplified RNA comprises unmodified nucleotides.
  5. The composition according to claim 3, wherein the self-amplified RNA comprises modified nucleotides.
  6. The composition according to claim 5, wherein less than approximately 50% of the nucleic acid in the self-amplifying RNA is modified nucleotides.
  7. The composition according to claim 1, wherein the first RNA molecule comprises a 5' cap; a 5' untranslated region (5'UTR); a coding region for a non-structural protein derived from an alphavirus; a first subgenome promoter derived from an alphavirus; a first open reading frame encoding a first target gene derived from influenza virus hemagglutinin (HA); a second subgenome promoter derived from an alphavirus; a second open reading frame encoding a second target gene derived from an influenza virus; a 3' untranslated region (3'UTR); and a 3' polyA sequence, wherein the second target gene derived from the influenza virus is an influenza non-structural (NS1) protein.
  8. The composition according to claim 7, further comprising an IRES sequence.
  9. The composition according to any one of claims 1 to 8, wherein the first RNA molecule and the second RNA molecule are not linked.
  10. The composition according to any one of claims 1 to 9, wherein the influenza NS1 is selected from the group consisting of influenza A virus NS1, influenza B virus NS1, influenza C virus NS1, and variants thereof.
  11. The composition according to any one of claims 1 to 10, wherein the influenza NS1 is selected from the group consisting of H1N1 NS1, H1N2 NS1, H2N2 NS1, H3N2 NS1, H5N1 NS1, H7N9 NS1, H7N7 NS1, H9N2 NS1, H7N2 NS1, H7N3 NS1, H5N2 NS1, H10N7 NS1, and any combination thereof.
  12. The composition according to any one of claims 1 to 11, wherein the influenza NS1 is encoded by an amino acid sequence having at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO: 33.
  13. The composition according to any one of claims 1 to 12, wherein the expression of the target mRNA is increased compared to the expression of the target mRNA in the absence of the nucleic acid molecule encoding the influenza NS1 protein.
  14. The composition according to any one of claims 1 to 13, wherein the expression of the target mRNA is increased by at least about 10% compared to the expression of the target mRNA in the absence of the nucleic acid molecule encoding the influenza NS1 protein.
  15. The composition according to any one of claims 1 to 14, wherein the increased expression of the target mRNA is sustained for at least about 6 hours.
  16. The composition according to any one of claims 1 to 15, wherein the RNA molecule is encapsulated in lipid nanoparticles.
  17. The immunogenic composition according to any one of claims 1 to 16.
  18. A method for expressing target mRNA in cells, comprising delivering a composition according to any one of claims 1 to 17 to the cells.
  19. A method for expressing a target or desired gene derived from an influenza virus in a subject requiring such expression, comprising administering the subject a composition according to any one of claims 1 to 18.

Description

Cross-reference to Applications This application claims the interests of U.S. Provisional Patent Application No. 63/484,186, filed 9 February 2023, U.S. Provisional Patent Application No. 63/484,747, filed 13 February 2023, and U.S. Provisional Patent Application No. 63/621,102, filed 15 January 2024, each of which is incorporated herein by reference in its entirety. This invention relates to compositions and methods for the preparation, manufacture, and therapeutic use of ribonucleic acid vaccines comprising polynucleotide molecules encoding one or more influenza antigens, such as hemagglutinin antigens. Influenza viruses belong to the Orthomyxoviridae family and are classified into three types (A, B, and C) based on differences in the antigenicity of their nucleoprotein (NP) and matrix (M) proteins. The genome of influenza A virus contains eight linear, negatively polarized, single-stranded RNA molecules (seven for influenza C virus), which encode several polypeptides, including RNA-dependent RNA polymerase proteins (PB2, PB1, and PA), nucleoproteins (NP) that form the nucleocapsid, matrix proteins (M1, M2, which are also surface-exposed proteins embedded in the viral membrane), two surface glycoproteins protruding from the lipoprotein envelope, hemagglutinin (HA) and neuraminidase (NA), and non-structural proteins (NS1 and NS2). Hemagglutinin is the major envelope glycoprotein of influenza A and B viruses, and the hemagglutinin-esterase (HE) of influenza C virus is homologous to HA. Challenges with conventional vaccines for treating and preventing influenza and other infectious diseases include the limited scope of vaccines, which provide protection only against closely related subtypes. Furthermore, the length of time required to complete current standard influenza virus vaccine manufacturing processes hinders the rapid development and production of suitable vaccines in pandemic situations. This graph shows functional anti-HA antibodies elicited by immunization of mice with LNP-formulated saRNA encoding influenza HA and/or NA, as measured by HAI. Figure 1A shows the results three weeks after priming. Female Balb/c mice were immunized on day 0 with 20 ng of LNP-formulated bicistronic saRNA vaccine preparation, 20 ng of LNP-formulated monocistronic saRNA vaccine preparation, a total of 40 ng (20 ng each) of a 1:1 mixture of saRNA-HA + saRNA-NA, and 200 ng of modRNA comparator encoding A/Wisconsin/588/2019 (H1N1) HA. Antibody response to A/Wisconsin/588/2019 was measured on day 21 (three weeks after immunization) by HAI or 1-Day MNT assay. HAI titers have been reported (geometric mean and geometric SD).This graph shows functional anti-HA antibodies elicited by immunization of mice with LNP-formulated saRNA encoding influenza HA and/or NA, as measured by HAI. Figure 1B shows the results two weeks after boost. Female Balb/c mice were immunized on day 0 with 20 ng of LNP-formulated bicistronic saRNA vaccine preparation, 20 ng of LNP-formulated monocistronic saRNA vaccine preparation, a total of 40 ng (20 ng each) of a 1:1 mixture of saRNA-HA + saRNA-NA, and 200 ng of modRNA comparator encoding A/Wisconsin/588/2019 (H1N1) HA. Antibody response to A/Wisconsin/588/2019 was measured on day 21 (three weeks after immunization) by HAI or 1-Day MNT assay. HAI titers have been reported (geometric mean and geometric SD).This graph shows the neutralizing antibodies elicited by immunization of mice with LNP-formulated saRNA encoding influenza HA and/or NA, as measured by 1-Day MNT. Figure 2A shows the results three weeks after priming. Female Balb/c mice were immunized on day 0 with 20 ng of LNP-formulated bicistronic saRNA vaccine preparation, 20 ng of LNP-formulated monocistronic saRNA vaccine preparation, a total of 40 ng (20 ng each) of a 1:1 mixture of saRNA-HA + saRNA-NA, and 200 ng of modRNA comparator encoding A/Wisconsin/588/2019 (H1N1) HA. Antibody responses to A/Wisconsin/588/2019 were measured on day 21 (three weeks after immunization) by HAI or 1-Day MNT assay. The 50% neutralizing titer has been reported (geometric mean and geometric SD).This graph shows the neutralizing antibodies elicited by immunization of mice with LNP-formulated saRNA encoding influenza HA and/or NA, as measured by 1-Day MNT. Figure 2B shows the results two weeks after boost. Female Balb/c mice were immunized on day 0 with 20 ng of LNP-formulated bicistronic saRNA vaccine preparation, 20 ng of LNP-formulated monocistronic saRNA vaccine preparation, a total of 40 ng (20 ng each) of a 1:1 mixture of saRNA-HA + saRNA-NA, and 200 ng of modRNA comparator encoding A/Wisconsin/588/2019 (H1N1) HA. Antibody responses to A/Wisconsin/588/2019 were measured on day 21 (three weeks after immunization) by HAI or 1-Day MNT assay. The 50% neutralizing titer has been reported (geometric mean and geometric SD).This graph shows the neutralizing antibodies extracted by immunization of mice with LNP-formulated saRNA encoding influenza HA and/or NA, as measu