CA2378866A1 - Fc fusion proteins for enhancing the immunogenicity of protein and peptide antigens - Google Patents

Fc fusion proteins for enhancing the immunogenicity of protein and peptide antigens Download PDF

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Publication number
CA2378866A1
CA2378866A1 CA002378866A CA2378866A CA2378866A1 CA 2378866 A1 CA2378866 A1 CA 2378866A1 CA 002378866 A CA002378866 A CA 002378866A CA 2378866 A CA2378866 A CA 2378866A CA 2378866 A1 CA2378866 A1 CA 2378866A1
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heavy chain
immunoglobulin heavy
chain constant
constant region
protein
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CA002378866A
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French (fr)
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CA2378866C (en
Inventor
Stephen D. Gillies
Kin Ming Lo
John S. Wesolowski, Jr.
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Merck Patent GmbH
Original Assignee
Stephen D. Gillies
Kin Ming Lo
John S. Wesolowski, Jr.
Lexigen Pharmaceuticals Corp.
Emd Lexigen Research Center Corp.
Merck Patent Gmbh
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Publication of CA2378866A1 publication Critical patent/CA2378866A1/en
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Publication of CA2378866C publication Critical patent/CA2378866C/en
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    • 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
    • A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
    • 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
    • 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
    • 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
    • A61K39/21—Retroviridae, e.g. equine infectious anemia virus
    • 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
    • A61K39/385—Haptens or antigens, bound to carriers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/475—Growth factors; Growth regulators
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/52—Cytokines; Lymphokines; Interferons
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/52—Cytokines; Lymphokines; Interferons
    • C07K14/53—Colony-stimulating factor [CSF]
    • C07K14/535—Granulocyte CSF; Granulocyte-macrophage CSF
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K19/00—Hybrid peptides, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
    • 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
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/62—DNA sequences coding for fusion proteins
    • 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
    • A61K2039/53—DNA (RNA) vaccination
    • 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
    • A61K2039/54—Medicinal preparations containing antigens or antibodies characterised by the route of administration
    • 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
    • A61K2039/545—Medicinal preparations containing antigens or antibodies characterised by the dose, timing or administration schedule
    • 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
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55511—Organic adjuvants
    • A61K2039/55522—Cytokines; Lymphokines; Interferons
    • 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
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55511—Organic adjuvants
    • A61K2039/55522—Cytokines; Lymphokines; Interferons
    • A61K2039/55527—Interleukins
    • A61K2039/55533—IL-2
    • 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
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55511—Organic adjuvants
    • A61K2039/55566—Emulsions, e.g. Freund's adjuvant, MF59
    • 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
    • A61K2039/60—Medicinal preparations containing antigens or antibodies characteristics by the carrier linked to the antigen
    • A61K2039/6031—Proteins
    • A61K2039/6056—Antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/02—Fusion polypeptide containing a localisation/targetting motif containing a signal sequence
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/40—Fusion polypeptide containing a tag for immunodetection, or an epitope for immunisation
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/70—Fusion polypeptide containing domain for protein-protein interaction
    • C07K2319/74—Fusion polypeptide containing domain for protein-protein interaction containing a fusion for binding to a cell surface receptor
    • C07K2319/75—Fusion polypeptide containing domain for protein-protein interaction containing a fusion for binding to a cell surface receptor containing a fusion for activation of a cell surface receptor, e.g. thrombopoeitin, NPY and other peptide hormones
    • 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
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/16011—Human Immunodeficiency Virus, HIV
    • C12N2740/16111—Human Immunodeficiency Virus, HIV concerning HIV env
    • C12N2740/16122—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
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/16011—Human Immunodeficiency Virus, HIV
    • C12N2740/16111—Human Immunodeficiency Virus, HIV concerning HIV env
    • C12N2740/16134—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S930/00—Peptide or protein sequence
    • Y10S930/01—Peptide or protein sequence
    • Y10S930/14—Lymphokine; related peptides
    • Y10S930/141—Interleukin

Abstract

Disclosed herein are methods and compositions for enhancing the immunogenici ty of a preselected protein or peptide antigen in a mammal. Immunogenicity is enhanced by fusing the preselected antigen to an immunoglobulin heavy chain constant region to produce an Fc-antigen fusion protein. The Fc-antigen fusi on proteins bind Fc receptors on the surface of antigen presenting cells, there by targeting the antigen to the antigen presenting cells in the mammal. In addition, disclosed is a family of adjuvants, for example, an Fc-adjuvant fusion protein, for use in combination with the Fc-antigen fusion proteins t o enhance or modulate a particular immune response against the preselected antigen.

Claims (45)

1. A method of enhancing the immunogenicity of a preselected antigen in a mammal, the method comprising:
administering to the mammal intramuscularly, intravenously, transdermally or subcutaneously, a fusion protein comprising an immunoglobulin heavy chain constant region linked by a polypeptide bond to the preselected antigen thereby to elicit an immune response against the preselected antigen, wherein the preselected antigen in the fusion protein elicits a stronger immune response in the mammal than the preselected antigen alone.
2. The method of claim 1, further comprising administering the fusion protein in combination with an adjuvant in an amount sufficient to enhance the immune response against the preselected antigen of the fusion protein relative to the immune response against the preselected antigen of the fusion protein administered without the adjuvant.
3. The method of claim 2, wherein the fusion protein and adjuvant are administered simultaneously.
4. The method of claim 2, wherein the adjuvant comprises a fusion protein comprising an immunoglobulin heavy chain constant region linked by a polypeptide bond to an adjuvant protein.
5. The method of claim 1 or 4, wherein the immunoglobulin heavy chain constant region comprises an immunoglobulin hinge region.
6. The method of claim 5, wherein the immunoglobulin heavy chain constant region comprises an immunoglobulin heavy chain constant region domain selected from the group consisting of a CH2 domain, a CH3 domain, and a CH4 domain.
7. The method of claim 5, wherein the immunoglobulin heavy chain constant region comprises a CH2 domain and a CH3 domain.
8. The method of claim 1 or 4, wherein the immunoglobulin heavy chain constant region is defined by an amino acid sequence corresponding to an amino acid sequence defining an immunoglobulin heavy chain constant region present in the same species as the mammal.
9. The method of claim 8, wherein the amino acid sequence defining the immunoglobulin heavy chain constant region corresponds to a human immunoglobulin heavy chain constant region.
10. The method of claim 1, wherein the preselected antigen is selected from the group consisting of a prostate-specific membrane antigen, an ectodomain of a cytokine receptor, a viral protein and a tumor-specific protein.
11. The method of claim 4, wherein the adjuvant protein is a cytokine.
12. The method of claim 11, wherein the cytokine is defined by an amino acid sequence corresponding to an amino acid sequence defining a cytokine present in the same species as the mammal.
13. The method of claim 12, wherein the cytokine is a human cytokine.
14. The method of claim 1, wherein the mammal is a human.
15. A composition for eliciting an immune response against a preselected antigen in a mammal, the composition comprising an admixture for intramuscular, intravenous, transdermal or subcutaneous administration selected from the group consisting of:
(a) an antigen fusion protein comprising an immunoglobulin heavy chain constant region linked by a polypeptide bond to the preselected antigen admixed with an adjuvant; and (b) a preselected antigen admixed with an adjuvant fusion protein comprising an immunoglobulin heavy chain constant region linked by a polypeptide bond to an adjuvant protein.
16. The composition of claim 15, wherein the adjuvant of clause (a) comprises a fusion protein comprising an immunoglobulin constant region linked by a polypeptide bond to an adjuvant protein.
17. The composition of claim 15, wherein the preselected antigen of clause (b) is linked by a polypeptide bond to an immunoglobulin heavy chain constant region.
18. The composition of claim 15, 16 or 17, wherein the immunoglobulin heavy chain constant region comprises an immunoglobulin hinge region.
19. The composition of claim 18, wherein the immunoglobulin heavy chain constant region comprises an immunoglobulin heavy chain constant region domain selected from the group consisting of a CH2 domain, a CH3 domain, and a CH4 domain.
20. The composition of claim 18, wherein the immunoglobulin heavy chain constant region comprises a CH2 domain and a CH3 domain.
21. The composition of claim 15, wherein the adjuvant of clause (a) comprises an oligonucleotide CpG sequence.
22. The composition of claim 15, wherein the preselected antigen is selected from the group consisting of a prostate-specific membrane antigen, an ectodomain of a cytokine receptor, a viral protein and a tumor-specific protein.
23. The composition of claim 15, wherein the antigen fusion protein of clause (a) or the adjuvant fusion of clause (b) is linked by a disulfide bond to a second immunoglobulin heavy chain constant region.
24. The composition of claim 15, wherein the adjuvant of clause (a) or the adjuvant protein of clause (b) is a cytokine.
25. The composition of claim 24, wherein the cytokine is a human cytokine.
26. The composition of claim 15, 16 or 17, wherein the immunoglobulin heavy chain constant region is defined by a amino acid sequence corresponding to an amino acid sequence defining a human immunoglobulin heavy chain constant region.
27. A method of enhancing the immunogenicity of a preselected antigen in a mammal, the method comprising:
administering to the mammal a nucleic acid sequence encoding a fusion protein comprising an immunoglobulin heavy chain constant region linked to the preselected antigen, whereupon expression of the nucleic acid sequence in the mammal results in the production of the fusion protein, the preselected antigen of which elicits a stronger immune response than the preselected antigen expressed from a nucleic acid encoding the preselected antigen alone.
28. The method of claim 27, wherein the nucleic acid encodes in a 5' to 3' direction the immunoglobulin heavy chain constant region and the preselected antigen.
29. The method of claim 28, wherein the immunoglobulin heavy chain constant region comprises an immunoglobulin hinge region.
30. The method of claim 27 or 29, wherein the immunoglobulin heavy chain constant region comprises a heavy chain domain selected from the group consisting of a domain, a CH3 domain, and a CH4 domain.
31. The method of claim 29, wherein the immunoglobulin heavy chain constant region comprises a CH2 domain and a CH3 domain.
32. The method of claim 27, wherein the preselected antigen is selected from the group consisting of a prostate-specific membrane antigen, an ectodomain of a cytokine receptor, a viral protein and an cancer-specific antigen.
33. The composition of claim 27, further comprising administering the nucleic acid sequence in combination with an adjuvant.
34. The method of claim 33, wherein the adjuvant comprises a nucleic acid sequence encoding an fusion protein comprising an immunoglobulin heavy chain constant region linked to an adjuvant protein.
35. A composition for eliciting an immune response against a preselected antigen in a mammal, the composition comprising:
(a) a first nucleic acid sequence encoding a fusion protein comprising an immunoglobulin heavy chain constant region and the preselected antigen, whereupon expression of the nucleic acid sequence in the mammal results in production of the fusion protein, the preselected antigen of which elicits a stronger immune response than the preselected antigen expressed from a nucleic acid encoding the preselected antigen alone;
and (b) an adjuvant.
36. The composition of claim 35, wherein the adjuvant comprises a second nucleic acid sequence encoding a fusion protein comprising an immunoglobulin heavy chain constant region linked by a peptide bond to an adjuvant protein.
37. The composition of claim 35 or 36, wherein the immunoglobulin heavy chain constant region comprises an immunoglobulin hinge region.
38. The composition of claim 35 or 36, wherein the immunoglobulin heavy chain constant comprises an immunoglobulin heavy chain domain selected from the group consisting of a CH2 domain, a CH3 domain, and a CH4 domain.
39. The composition of claim 37, wherein the immunoglobulin heavy chain constant comprises an immunoglobulin heavy chain domain selected from the group consisting of a CH2 domain, a CH3 domain, and a CH4 domain.
40. The composition of claim 35, wherein the preselected antigen is selected from the group consisting of a prostate-specific membrane antigen, an ectodomain of a cytokine receptor, a viral protein and a cancer-specific antigen.
41. The composition of claim 36, wherein the adjuvant protein is a cytokine.
42. The composition of claim 35, wherein the first nucleic acid sequence is operably disposed within a replicable expression vector.
43. The composition of claim 36, wherein the second nucleic acid sequence is operably disposed within a replicable expression vector.
44. A method for enhancing the immunogenicity of a preselected antigen in a mammal, the method comprising:
administering to a mammal, either simultaneously or sequentially, a first fusion protein comprising an antigen protein with a localizing protein, and a second fusion protein comprising an adjuvant protein and said localizing protein, said localizing protein causing an increase in concentration of said first and second fusion proteins in a region of the mammal accessible to the immune system.
45. A method for enhancing the immunogenicity of a preselected antigen in a mammal, the method comprising:

administering to a mammal a fusion protein comprising an antigen protein, an adjuvant protein and a localizing protein, said localizing protein causing an increase in concentration of said antigen and adjuvant proteins in a region of the mammal accessible to the immune system.
CA2378866A 1999-07-21 2000-07-21 Fc fusion proteins for enhancing the immunogenicity of protein and peptide antigens Expired - Fee Related CA2378866C (en)

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US14496599P 1999-07-21 1999-07-21
US60/144,965 1999-07-21
PCT/US2000/019816 WO2001007081A1 (en) 1999-07-21 2000-07-21 Fc fusion proteins for enhancing the immunogenicity of protein and peptide antigens

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CA2378866C CA2378866C (en) 2011-06-07

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