Nucleic acid vaccines

• US 10,022,435 B2
• Filed: 04/01/2016
• Issued: 07/17/2018
• Est. Priority Date: 04/23/2014
• Status: Active Grant

• Current Assignee: Moderna TX, Inc. (Moderna, Inc.)
• Original Assignee: Moderna TX, Inc. (Moderna, Inc.)
• Inventors: Ciaramella, Giuseppe, Bouchon, Axel, Huang, Eric Yi-Chun
• Primary Examiner(s): Blumel, Benjamin P
• Application Number:
  US15/089,050
• Publication Number:
  US 20160317647A1
• Time in Patent Office: 837 Days
• Field of Search: None
• CPC Class Codes: A61K 2039/53 DNA (RNA) vaccination
• CPC Class Codes: A61K 2039/545 characterised by the dose, timing or administration schedule
• CPC Class Codes: A61K 2039/55555 Liposomes; Vesicles, e.g. nanoparticles; Spheres, e.g. nanospheres; Polymers
• CPC Class Codes: A61K 31/7105 Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
• CPC Class Codes: A61K 39/12 Viral antigens
• CPC Class Codes: A61K 39/145 Orthomyxoviridae, e.g. influenza virus
• CPC Class Codes: A61K 39/155 Paramyxoviridae, e.g. parainfluenza virus
• CPC Class Codes: A61K 39/39 characterised by the immunostimulating additives, e.g. chemical adjuvants
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61K 9/1271 Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers liposome as conjugate A61K47/6911
• CPC Class Codes: A61K 9/5123 Organic compounds, e.g. fats, sugars
• CPC Class Codes: A61K 9/5146 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, polyamines, polyanhydrides
• CPC Class Codes: A61P 31/16 for influenza or rhinoviruses
• CPC Class Codes: C12N 2760/16122 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2760/16134 Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• CPC Class Codes: C12N 2760/16234 Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• CPC Class Codes: C12N 7/00 Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The invention relates to compositions and methods for the preparation, manufacture and therapeutic use ribonucleic acid vaccines (NAVs) comprising polynucleotide molecules encoding one or more antigens.

Claims

1. A method of vaccinating a subject comprising administering to the subject a nucleic acid vaccine comprising one or more RNA polynucleotides comprising an open reading frame encoding an antigenic polypeptide that is derived from an infectious agent, i) wherein the RNA polynucleotide does not include a stabilization element, or wherein the nucleic acid vaccine is not co-formulated or or co-administered with an adjuvant and ii) is formulated within a cationic lipid nanoparticle having a molar ratio of about 20-60% ionizable cationic lipid:

about 5-25% non-cationic lipid;

about 25-55% sterol; and

about 0.5-15% PEG-modified lipid, wherein the nucleic acid vaccine elicits an immune response having a longer lasting antibody titer than an antibody titer elicited by a reference nucleic acid vaccine comprising one or more RNA polynucleotides comprising an open reading frame encoding an antigenic polypeptide that is derived from an infectious agent i) wherein the RNA polynucleotide does include a stabilization element or wherein the nucleic acid vaccine is co-formulated or co-administered with an adjuvant and is ii) not formulated within a cationic lipid nanoparticle having a molar ratio of about 20-60% ionizable cationic lipid;

about 5-25% non-cationic lipid;

about 25-55% sterol; and

about 0.5-15% PEG-modified lipid.

2. The method of claim 1, wherein the nucleic acid vaccine is formulated to produce neutralizing antibodies within one week of a single administration.

4. The method of claim 1, wherein the RNA polynucleotide is present in a dosage of between 25 and 100 micrograms.

5. The method of claim 1, wherein the method comprises administering to the subject a single dosage of between 0.001 mg/kg and 0.005 mg/kg of the nucleic acid vaccine in an effective amount to vaccinate the subject.

6. The method of claim 1, wherein the nucleic acid vaccine is administered to the subject by intradermal or intramuscular injection.

7. The method of claim 1, wherein the ionizable cationic lipid nanoparticle comprises a molar ratio of about 50% cationic lipid, about 1.5% PEG-modified lipid, about 38.5% cholesterol and about 10% non-cationic lipid.

8. The method of claim 1, wherein the cationic lipid nanoparticle comprises a molar ratio of about 55% cationic lipid, about 2.5% PEG lipid, about 32.5% cholesterol and about 10% non-cationic lipid.

9. The method of claim 1, wherein the open reading frame is codon-optimized.

10. The method of claim 1, wherein the ionizable cationic lipid nanoparticle has a polydispersity value of less than 0.4.

11. The method of claim 1, wherein the polynucleotide has a poly-A tail of 80-250 nucleotides in length.

12. The method of claim 1, wherein the polynucleotide comprises at least one 5′

terminal cap and at least one chemical modification.

13. The method of claim 1, wherein a second dose of the nucleic acid vaccine is administered to the subject.

14. The method of claim 1, wherein the RNA polynucleotide accumulates at a 100 fold higher level in the local lymph node in comparison with the distal lymph node.

15. The method of claim 1, wherein the RNA polynucleotide includes a chemical modification selected from the group consisting of pseudouridine, N1-methylpseudouridine, 2-thiouridine, 4′

-thiouridine, 5-methylcytosine, 2-thio-1-methyl-1-deaza-pseudouridine, 2-thio-1-methyl-pseudouridine, 2-thio-5-aza-uridine, 2-thio-dihydropseudouridine, 2-thio-dihydrouridine, 2-thio-pseudouridine, 4-methoxy-2-thio-pseudouridine, 4-methoxy-pseudouridine, 4-thio-1-methyl-pseudouridine, 4-thio-pseudouridine, 5-aza-uridine, dihydropseudouridine, 5-methoxyuridine, and 2′

-O-methyl uridine.

3. A method of vaccinating a subject comprising administering to the subject a nucleic acid vaccine comprising one or more RNA polynucleotides comprising an open reading frame encoding an antigenic polypeptide that is derived from an infectious agent, i) wherein the RNA polynucleotide does not include a stabilization element or wherein the nucleic acid vaccine is not co-formulated or or co-administered with an adjuvant and ii) is formulated within a cationic lipid nanoparticle having a molar ratio of about 20-60% ionizable cationic lipid:

about 5-25% non-cationic lipid;

about 25-55% sterol; and

about 0.5-15% PEG-modified lipid, wherein a level of antigen expression produced in the subject after administration of the nucleic acid vaccine exceeds a level of antigen expression produced by a reference nucleic acid vaccine comprising one or more RNA polynucleotides comprising an open reading frame encoding an antigenic polypeptide that is derived from an infectious agent i) wherein the RNA polynucleotide does include a stabilization element or wherein the nucleic acid vaccine is co-formulated or co-administered with an adjuvant and is ii) not formulated within a cationic lipid nanoparticle having a molar ratio of about 20-60% ionizable cationic lipid;

about 5-25% non-cationic lipid;

about 25-55% sterol; and

about 0.5-15% PEG-modified lipid.

16. The method of claim 3, wherein the RNA polynucleotide is present in a dosage of between 25 and 100 micrograms.

17. The method of claim 3, wherein the method comprises administering to the subject a single dosage of between 0.001 mg/kg and 0.005 mg/kg of the nucleic acid vaccine in an effective amount to vaccinate the subject.

18. The method of claim 3, wherein the nucleic acid vaccine is administered to the subject by intradermal or intramuscular injection.

19. The method of claim 3, wherein the ionizable cationic lipid nanoparticle comprises a molar ratio of about 50% cationic lipid, about 1.5% PEG-modified lipid, about 38.5% cholesterol and about 10% non-cationic lipid.

20. The method of claim 3, wherein the cationic lipid nanoparticle comprises a molar ratio of about 55% cationic lipid, about 2.5% PEG lipid, about 32.5% cholesterol and about 10% non-cationic lipid.

21. The method of claim 3, wherein the open reading frame is codon-optimized.

22. The method of claim 3, wherein the ionizable cationic lipid nanoparticle has a polydispersity value of less than 0.4.

23. The method of claim 3, wherein the polynucleotide has a poly-A tail of 80-250 nucleotides in length.

24. The method of claim 3, wherein the polynucleotide comprises at least one 5′

terminal cap and at least one chemical modification.

25. The method of claim 3, wherein a second dose of the nucleic acid vaccine is administered to the subject.

26. The method of claim 3, wherein the RNA polynucleotide accumulates at a 100 fold higher level in the local lymph node in comparison with the distal lymph node.

27. The method of claim 3, wherein the RNA polynucleotide includes a chemical modification selected from the group consisting of pseudouridine, N1-methylpseudouridine, 2-thiouridine, 4′

-thiouridine, 5-methylcytosine, 2-thio-1-methyl-1-deaza-pseudouridine, 2-thio-1-methyl-pseudouridine, 2-thio-5-aza-uridine, 2-thio-dihydropseudouridine, 2-thio-dihydrouridine, 2-thio-pseudouridine, 4-methoxy-2-thio-pseudouridine, 4-methoxy-pseudouridine, 4-thio-1-methyl-pseudouridine, 4-thio-pseudouridine, 5-aza-uridine, dihydropseudouridine, 5-methoxyuridine, and 2′

-O-methyl uridine.