Classifications

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  • 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
  • A61K39/155—Paramyxoviridae, e.g. parainfluenza virus
• • A—HUMAN NECESSITIES
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• • A—HUMAN NECESSITIES
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  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
  • A61P31/12—Antivirals
  • A61P31/14—Antivirals for RNA viruses
• • 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
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  • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
  • A61K2039/525—Virus
  • A61K2039/5258—Virus-like particles
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  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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  • 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
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  • 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
  • A61K2039/55—Medicinal preparations containing antigens or antibodies characterised by the host/recipient, e.g. newborn with maternal 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
  • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
  • A61K2039/55505—Inorganic adjuvants
• • 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
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  • A61K2039/57—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2
  • A61K2039/575—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2 humoral response
• • 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
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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  • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
  • C12N2760/00011—Details
  • C12N2760/18011—Paramyxoviridae
  • C12N2760/18511—Pneumovirus, e.g. human respiratory syncytial virus
  • C12N2760/18522—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
  • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
  • C12N2760/00011—Details
  • C12N2760/18011—Paramyxoviridae
  • C12N2760/18511—Pneumovirus, e.g. human respiratory syncytial virus
  • C12N2760/18523—Virus like particles [VLP]
• • 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
  • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
  • C12N2760/00011—Details
  • C12N2760/18011—Paramyxoviridae
  • C12N2760/18511—Pneumovirus, e.g. human respiratory syncytial virus
  • C12N2760/18534—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein

Definitions

• the present disclosure relates to vaccines for Respiratory Syncytial Virus (RSV).
• RSV Respiratory Syncytial Virus
• Respiratory syncytial virus is a single-strand, negative sense RNA virus of the family Pneumo viridae.
• RSV circulates seasonally and is a major cause of lower respiratory tract infection (LRTI) worldwide.
• LRTI lower respiratory tract infection
• Epidemiological data suggest that in the US alone RSV may cause >170,000 hospitalizations and -14,000 deaths annually (Colosia et al., PLoS One. 2017;12(8):e0182321).
• RSV is an important cause of respiratory disease in Europe (Broberg etal., Euro Surveill. 2018;23(5):17-00284).
• RSV infections are a major cause of bronchiolitis and pneumonia in young children globally (Nair et al., Lancet. 2010; 375(9725): 1545-55).
• LRTI lower respiratory tract infection
• RSV is the single most important viral LRTI during infancy and early childhood worldwide, and particularly in preterm infants and in infants with cardiopulmonary conditions, who are considered at high risk for complications and hospitalization.
• ARI acute respiratory illness
• RSV-A and/or RSV-B Treatment for illness caused by RSV (e.g., RSV-A and/or RSV-B) is mainly symptomatic and prevention consists mainly of infection control strategies, such as hand washing and droplet precautions.
• Neonates at high risk for RSV infection such as premature infants or infants with cardiopulmonary disease, are candidates for prophylaxis with humanized monoclonal antibody palivizumab (Resch et al., Hum Vaccin Immunother. 2017;13(9):2138-2149), which has been shown to be only moderately effective (45-55%) in reducing hospitalization for RSV (Ambrose etal., Human Vaccines & Immunotherapeutics. 2014;10:10, 2785-2788).
• Antiviral agents forthe prevention and treatment of RSV infections in elderly adults are currently not available, and there is no vaccine licensed for prevention of disease due to RSV infections.
• RSV F protein is a major conserved surface antigen of RSV and antibodies against it are associated with protection against disease.
• RSV F-protein is a validated target for protection against infection by RSV as demonstrated by the clinical efficacy of palivizumab, a monoclonal antibody that binds F-antigen and leads to neutralization of the virus (Johnson et al., J Infect Dis. 1997 Nov;176(5):1215-24).
• RSV-F protein is known to undergo a significant change in structure from prefusion to postfusion form which catalyzes viral and host membrane fusion to allow for viral entry into the cell (McLellan et al., Science. 2013;342(6158):592-8).
• Prefusion F-protein has important epitopes that are lost during the transition to postfusion F-protein (Melero et al., Vaccine. 2017;35(3):461-468).
• Antibody depletion studies with human sera absorbed with RSV F protein in either conformation demonstrate that the majority of the neutralizing response against RSV F protein targets the prefusion structure (Krarup et al., Nat Commun. 2015;6:8143). These studies also demonstrate the potential for antibodies that bind postfusion F protein to interfere with neutralization (Ngwuta et al., Sci Transl Med. 2015;7(309):309ral62). In general, high levels of antibodies against RSV F protein are associated with protection against severe disease. However, generating high-titers of neutralizing antibodies against RSV F protein remains challenging, due to the specific biochemical nature of the RSV F protein and the unpredictability of vaccine responses to RSV F.
• a pharmaceutical composition comprising a protein complex comprising a first component comprising an RSV F protein and a first multimerization domain, and a second component comprising a second multimerization domain; and one or more pharmaceutically acceptable diluents or excipients.
• the pharmaceutical composition comprises an oil-in-water adjuvant.
• the pharmaceutical composition comprises an aluminum hydroxide-adjuvant.
• the protein complex is an icosahedral protein complex.
• the protein complex comprises 20 copies of the first component and 12 copies of the second component.
• the RSV F protein comprises an amino acid sequence which is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence of any one of SEQ ID NOs: 14, 34, and 35.
• the first multimerization domain comprises an amino acid sequence which is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence of any one of SEQ ID NOs: 24 and 30-31; and/or the second multimerization domain comprises an amino acid sequence which is at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 100% identical to an amino acid sequence selected from any one of SEQ ID NOS: 22-23, 25-29, and 32.
• the first component comprises an amino acid sequence which is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 6; and the second component comprises an amino acid sequence which is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 26.
• a unit dose of the pharmaceutical composition comprising about 0.5 ⁇ g to about 1 ⁇ g, about 20 ⁇ g to about 25 ⁇ g, about 70 ⁇ g to about 75 ⁇ g, about 100 ⁇ g to about 125 ⁇ g, or about 200 ⁇ g to about 250 ⁇ g of the protein complex.
• a method of vaccinating a subject comprising administering to the subject an effective amount of a pharmaceutical composition provided herein.
• a method of generating an immune response in a subject comprising administering to the subject an effective amount of a pharmaceutical composition provided herein.
• provided herein is a method of preventing RSV disease in a subject, comprising administering to the subject an effective amount of a pharmaceutical composition provided herein.
• the subject is at risk of severe RSV disease.
• the subject is an adult of over 60 years of age. In some embodiments, the subject is a healthy adult of 18-45 years of age.
• a method of generating an immune response in an unborn child comprising administering an effective amount of a pharmaceutical composition provided herein to the mother of said unborn child.
• the pharmaceutical composition is administered to the mother in the last trimester of the pregnancy.
• an effective amount of a pharmaceutical composition comprises about 0.5 ⁇ g to about 1 ⁇ g, about 20 ⁇ g to about 25 ⁇ g, about 70 ⁇ g to about 75 ⁇ g, about 100 ⁇ g to about 125 ⁇ g, or about 200 ⁇ g to about 250 ⁇ g of the protein complex.
• a method provided herein further comprising administering a second dose of a pharmaceutical composition provided herein.
• the second dose is administered within about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks, about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 9 months, or about 12 months of the first dose.
• the method provided herein further comprises administering a third dose of a pharmaceutical composition provided herein.
• the third dose is administered about 1 year, about 2 years, about 3 years, about 4 years, or about 5 years after the second dose.
• a method provided herein further comprises administering subsequent doses at regular intervals of about 1, 2, 3, 4 or 5 years.
• a method provided herein limits the development of an RSV infection in a subject.
• the method results in the production of RSV- A-specific neutralizing antibodies in the subject.
• the method results in an increase in RSV-A-specific neutralizing antibodies in the subject of at least about 3- fold, at least about 4-fold, at least about 5-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, or at least about 25-fold compared to baseline.
• the increase in RSV-A-specific neutralizing antibodies is detectable within about one week, within about 2 weeks, within about 3 weeks, within about 4 weeks, within about 5 weeks, within about 6 weeks, within about 7 weeks, within about 8 weeks, within about 9 weeks, within about 10 weeks, within about 11 weeks, or within about 12 weeks of administration of the pharmaceutical composition.
• the method results in the production of RSV-B-specific neutralizing antibodies in the subject.
• the method results in an increase in RSV-B-specific neutralizing antibodies in the subject of at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, or at least about 25-fold compared to baseline.
• the increase in RSV-B-specific neutralizing antibodies is detectable within about one week, within about 2 weeks, within about 3 weeks, within about 4 weeks, within about 5 weeks, within about 6 weeks, within about 7 weeks, within about 8 weeks, within about 9 weeks, within about 10 weeks, within about 11 weeks, or within about 12 weeks of administration of the pharmaceutical composition.
• the method results in the production of RSV F-protein-specific IgG antibodies in the subject. In some embodiments, the method results in an increase of RSV F-protein-specific neutralizing antibodies in the subject of at least about 3- fold, at least about 4-fold, at least about 5-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, or at least about 25-fold compared to baseline.
• the increase in RSV F-protein-specific IgG antibodies is detectable within about one week, within about 2 weeks, within about 3 weeks, within about 4 weeks, within about 5 weeks, within about 6 weeks, within about 7 weeks, within about 8 weeks, within about 9 weeks, within about 10 weeks, within about 11 weeks, or within about 12 weeks of administration of the pharmaceutical composition.
• the method results in the production of RSV F-protein-specific memory-B-cells in the subject. In some embodiments, the method results in an increase in RSV F-protein-specific memory-B-cells in the subject of at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, or at least about 25-fold compared to baseline.
• the increase in RSV F-protein-specific memory-B-cells is detectable within about one week, within about 2 weeks, within about 3 weeks, within about 4 weeks, within about 5 weeks, within about 6 weeks, within about 7 weeks, within about 8 weeks, within about 9 weeks, within about 10 weeks, within about 11 weeks, or within about 12 weeks of administration of the pharmaceutical composition.
• the method results in the production of RSV F-protein-specific T-cells in the subject. In some embodiments, the method results in an increase in RSV F-protein-specific T-cells in the subject of at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, or at least about 25-fold compared to baseline.
• the increase in RSV F- protein-specific T-cells is detectable within about one week, within about 2 weeks, within about 3 weeks, within about 4 weeks, within about 5 weeks, within about 6 weeks, within about 7 weeks, within about 8 weeks, within about 9 weeks, within about 10 weeks, within about 11 weeks, or within about 12 weeks of administration of the pharmaceutical composition.
• FIGs. 1A-1C shows neutralization antibody titers measured in naive mice on Days 0 (FIG. 1A), 42 (FIG. IB) and 56 (FIG. 1C) after administration of RSV vaccine (two- tailed, unpaired t test).
• Group 1 RSV vaccine (8.33 ⁇ g);
• Group 2 RSV vaccine (8.33 ⁇ g) plus Alhydrogel;
• Group 3 RSV vaccine (2.5 ⁇ g);
• Group 4 RSV vaccine (2.5 ⁇ g) plus Alhydrogel;
• Group 5 RSV vaccine (0.83 ⁇ g);
• Group 6 RSV vaccine (0.83 ⁇ g) plus Alhydrogel;
• Group 7 RSV vaccine (8.33 ⁇ g) plus Addavax;
• Group 8 control sera.
• FIG. 2 shows the increase in neutralizing antibody titers with administration of Alhydrogel-adjuvanted or non-adjuvated RSV vaccine or RSV F-protein in RSV-primed mice.
• FIG. 3 shows RSV neutralizing antibody titers in rabbits; impact of preimmunization with VLP core.
• Group 1 5 New Zealand White (NZW) female rabbits were administered with VLP core plus Addavax (oil-in-water emulsion) on Days 1 and 14. Subsequently, the rabbits were vaccinated with RsV vaccine plus Alhydrogel (aluminum hydroxide adjuvant) on Days 56, 70 and 84.
• Group 2 NZW 3 female rabbits each were vaccinated with RSV vaccine without Alhydrogel and no VLP core prior administration.
• Group 3 NZW 3 female rabbits each were vaccinated with RSV vaccine with Alhydrogel and no VLP core prior administration.
• FIG. 4 is the study design for a Phase 1/lb randomized, observer-blinded, placebo- controlled study to evaluate IVX-121 administration in young and older adult subjects.
• FIG. 5 shows a summary of safety data. There were no serious adverse events (SAEs), AEs of special interest (AESIs) or adverse events (AEs) leading to study withdrawal.
• SAEs serious adverse events
• AESIs AEs of special interest
• AEs adverse events
• FIG. 7 shows a graph of RSV-A neutralizing antibodies (nAB). Geometric mean titer (GMT) is expressed in international units per milliliter (lU/mL). GMT of unadj uvanted IVX-121 are comparable in young and older adults.
• FIG. 8 shows a graph of RSV- A neutralizing antibodies (nAB), unadjuvanted versus adj uvanted.
• Geometric mean titer (GMT) is expressed in international units per milliliter (lU/mL).
• Alum adjuvant had no beneficial effect in young and older adults.
• FIG. 9 shows tables summarizing neutralizing and binding antibody data.
• compositions comprising a protein complex which may be used to vaccinate against RSV (e.g., RSV-A subtype and/or RSV-B subtype).
• a composition comprising a non-replicating recombinant protein-based vaccine which is presented to the immune system as a virus-like particle (VLP).
• VLPs virus-like particle
• Several naturally occurring VLPs are constituents of licensed vaccines (e.g., hepatitis B, human papilloma virus) and have been safely used across all age groups, ranging from young children to older adults. Without wishing to be bound by theory, it is thought that such a vaccine can boost RSV-neutralizing antibody titers while reducing the induction of binding, non-neutralizing antibodies which were previously associated with enhanced respiratory disease (ERD).
• ETD enhanced respiratory disease
• the term “about” is used to indicate that a value includes the inherent variation of error for the device or the method being employed to determine the value, or the variation that exists among the samples being measured. Unless otherwise stated or otherwise evident from the context, the term “about” means within 10% above or below the reported numerical value (except where such number would exceed 100% of a possible value or go below 0%). When used in conjunction with a range or series of values, the term “about” applies to the endpoints of the range or each of the values enumerated in the series, unless otherwise indicated. As used in this application, the terms “about” and “approximately” are used as equivalents.
• sequence identity refers to the extent to which two optimally aligned polynucleotides or polypeptide sequences are invariant throughout a window of alignment of residues, e.g. nucleotides or amino acids.
• An “identity fraction” for aligned segments of a test sequence and a reference sequence is the number of identical residues which are shared by the two aligned sequences divided by the total number of residues in the reference sequence segment, i.e. the entire reference sequence or a smaller defined part of the reference sequence. “Percent identity” is the identity fraction times 100.
• sequence identity refers to sequence identity calculated as percentage of exact matches between the reference sequence and the sequence of interest, across the entire length of the reference sequence, when the sequence of interest is aligned to the reference sequence using the Blast-p program of the National Center for Biotechnology Information (NCBI) online alignment tool, version 2.11.0 (released October 19, 2020), Altschul et al. J. Mol. Biol. 215:403-410 (1990).
• NCBI National Center for Biotechnology Information
• heterologous vaccine and “heterologous vaccination” refer to a vaccine given to a subject who has received or will receive a vaccination for the same indication (e.g., RSV) using a vaccine made with another technology (e.g., an mRNA vaccine, adenoviral vector vaccine, or a protein subunit vaccine).
• a “heterologous vaccine” refers to a vaccine made using a different technology type than the reference vaccine.
• a “heterologous boost” or “heterologous boost vaccine” refers to a heterologous vaccine (e.g., a protein-based VLPs) given to a subject who has received a vaccination for the same indication (e.g., RSV) using a vaccine made with another technology (e.g., an mRNA vaccine, adenoviral vector vaccine, or a protein subunit vaccine).
• a heterologous vaccine e.g., a protein-based VLPs
• a vaccine made with another technology e.g., an mRNA vaccine, adenoviral vector vaccine, or a protein subunit vaccine.
• the term “prime vaccine” refers to the first vaccine in a vaccination protocol or to a first set of vaccines administered prior to a heterologous boost vaccine.
• an mRNA vaccine or adenoviral vaccine may be administered first, optionally followed by a second prime vaccine after a suitable interval, and then the heterologous vaccine may be administered.
• the heterologous vaccine may serve to “boost” the immune response to the prime vaccine.
• a “priming vaccine” as used herein refers to a vaccine comprising an agent(s) that encodes the target antigen to which an immune response is to be generated. Priming vaccines are administered to the subject in an amount effective to elicit an immune response to the target antigen.
• a “heterologous prime-boost vaccination” refers to a vaccine given to a subject who will receive a vaccination for the same indication (e.g., RSV) using a vaccine made with another technology.
• the initial dose (primary vaccine or prime vaccination) of a vaccine may an mRNA vaccine (or alternatively, the subject may have been diagnosed with the indication, e.g., RSV), and subsequently receive a second vaccination for the same indication, wherein the second vaccination is of a different technology - a heterologous vaccination (e.g., a protein-based VLP).
• heterologous prime-boost vaccination includes a primary vaccination for an indication, and a subsequent vaccination for the same indication, wherein the heterologous vaccination is administered 3 months to 6 months after the heterologous prime vaccine, or 4 or more months after a heterologous prime vaccine, or 6 months or more after a heterologous prime vaccine, or 10 months or more after a heterologous prime vaccine.
• the heterologous boost vaccination is administered 1 year after a heterologous prime vaccine.
• heterologous prime or “heterologous prime vaccine” refers to a vaccine given to a subject who will receive a vaccination for the same indication (e.g., RSV) using a vaccine made with another technology (e.g., an mRNA vaccine, adenoviral vector vaccine, or a protein subunit vaccine).
• a vaccine made with another technology e.g., an mRNA vaccine, adenoviral vector vaccine, or a protein subunit vaccine.
• virus-like particle refers to a molecular assembly that resembles a virus, but is non-infectious, and that displays an antigenic protein, or antigenic fragment thereof, of a viral protein or glycoprotein.
• a “protein-based VLP” refers to a VLP formed from proteins or glycoproteins and substantially free of other components (e.g., lipids). Protein-based VLPs may include post-translation modification and chemical modification, but are to be distinguished from micellar VLPs and VLPs formed by extraction of viral proteins from live or live inactivated virus preparations.
• the term “designed VLP” refers to a VLP comprising one or more polypeptides generated by computational protein design.
• symmetric VLP refers to a protein-based VLP with a symmetric core. These include but are not limited to designed VLPs.
• the protein ferritin has been used to generate a symmetric, protein-based VLP using naturally occurring ferritin sequences.
• Ferritin-based VLP are distinguished from designed VLPs in that no protein engineering is necessary to form a symmetric VLP from ferritin, other than fusing the viral protein to the ferritin molecule.
• Protein design methods can be used to generate similar one- and two-component nanostructures based on template structures (e.g., structures deposited in the Protein Data Bank) or de novo (i.e., by computational design of new proteins having a desired structure but little or no homology to naturally occurring proteins).
• Such one- and two-component nanostructures can then be used as the core of a designed VLP.
• the terms “protein nanoparticle” or “nanoparticle” and the term “nanostructure” may be used to refer to protein-based VLPs as described herein.
• an “immunogenic composition” is a composition that comprises an antigen where administration of the composition to a subject results in the development in the subject of a humoral and/or a cellular immune response to the antigen.
• the term “subject” includes humans and other animals. Typically, the subject is a human. For example, the subject may be an adult, a teenager, a child (2 years to 14 years of age), an infant (birth to 2 year), or a neonate (up to 2 months). In particular, aspects, the subject is up to 4 months old, or up to 6 months old.
• the adults are older adults, such as older adults over 50 years of age, older adults over 55 years of age, older adults over 60 years of age, older adults over 65 years of age.
• the subject is a pregnant woman or a woman intending to become pregnant.
• subject is not a human; for example a non-human primate; for example, a baboon, a chimpanzee, a gorilla, or a macaque.
• the subject may be a pet, such as a dog or cat.
• the present disclosure relates generally to vaccination of a subject with a protein complex comprising a first component comprising an RSV F protein and a first multimerization domain.
• the protein complex may comprise the F protein of RSV-A or RSV-B. Illustrative sequences of the F proteins of RSV-A and B are set forth in SEQ ID NOs: 14 and 34, respectively.
• the F protein portion and the first multimerization domain may be linked by any suitable means, including co-expression as a fusion protein.
• the protein complex may optionally comprise a second component comprising a second multimerization domain.
• the pharmaceutical composition typically comprises one or more pharmaceutically acceptable diluents or excipients.
• the protein complex is a nanostructure, nanoparticle, or protein-based virus-like particle.
• the protein complex is an icosahedral protein complex, such as those disclosed in U.S. Patent No. 10,248,758 or U.S. Patent Pub. No. 2020/0392187 Al, the contents of which are incorporated by reference herein in their entireties.
• the multimerization domains may be derived from a naturally-occurring protein sequence by substitution of at least one amino acid residue or by additional at the N- or C- terminus of one or more residues.
• the first multimerization domain comprises a protein sequence determined by computational methods. This first multimerization domain may form the entire core of the VLP; or the core of the VLP may comprise one or more additional polypeptides (also referred to a “second component” or third, fourth, fifth component and so on), such that the VLP comprises two, three, four, five, six, seven, or more multimerization domains.
• the first component will form trimers related by 3-fold rotational symmetry and the second component will form pentamers related by 5- fold rotational symmetry.
• the VLP forms an “icosahedral particle” having 153 symmetry.
• these one or more pluralities of component may be arranged such that the members of each plurality of component are related to one another by symmetry operators.
• a general computational method for designing self-assembling protein materials, involving symmetrical docking of protein building blocks in a target symmetric architecture, is disclosed in U.S. Patent Pub. No. US 2015/0356240 Al.
• the “core” of the VLP is used herein to describe the central portion of the VLP that links together the several copies of the RSV F protein ectodomain, or antigenic fragments thereof, displayed by the VLP.
• the first component comprises a first polypeptide comprising an F protein, a linker, and a multimerization domain.
• the VLP is adapted to display the F protein from two or more diverse strains of RSV.
• the same VLP displays mixed populations of protein antigens or mixed heterotrimers of protein antigens from different strains of RSV.
• sequences of the F protein of various RSV strains are known in the art, see, e.g., NCBI Accession Nos.: QFX69124.1, QFX69112.1, APW78900.1, APW78889.1, APW78878.1, APW78867.1, APW78856.1, APW78845.1, APW78834.1, APW78823.1, APW78812.1, APW78801.1, APW78790.1, APW78779.1, APW78768.1, APW78757.1, APW78746.1, APW78735.1, APW78724.1, APW78713.1, APW78702.1, APW78691.1, APW78680.1, APW78669.1, APW78658.1, APW78647.1, APW78636.1, APW78625.1, APW78614.1, and AAR14266.1.
• VLPs of the present disclosure display antigenic proteins in various ways including as gene fusion or by other means disclosed herein.
• “linked to” or “attached to” denotes any means known in the art for causing two polypeptides to associate.
• the association may be direct or indirect, reversible or irreversible, weak or strong, covalent or non-covalent, and selective or nonselective.
• attachment is achieved by genetic engineering to create an N- or C-terminus fusion of an antigen to one of the pluralities of polypeptides composing the VLP.
• the VLP may consist of, or consist essentially of, one, two, three, four, five, six, seven, eight, nine, or ten pluralities of polypeptides displaying one, two, three, four, five, six, seven, eight, nine, or ten pluralities of antigens, where at least one of the pluralities of antigen is genetically fused to at least one of the plurality of polypeptides.
• the VLP consists essentially of one plurality of polypeptides capable of self-assembly and comprising the plurality of antigenic proteins genetically fused thereto. In some cases, the VLP consists essentially of a first plurality of polypeptides comprising a plurality of antigens; and a second plurality of polypeptides capable of co-assembling into two- component VLP, one plurality of polypeptides linking the antigenic protein to the VLP and the other plurality of polypeptides promoting self-assembly of the VLP.
• attachment is achieved by post-translational covalent attachment between one or more pluralities of polypeptides and one or more pluralities of antigenic protein.
• chemical cross-linking is used to non-specifically attach the antigen to a VLP polypeptide.
• chemical cross-linking is used to specifically attach the antigenic protein to a VLP polypeptide (e.g. to the first polypeptide or the second polypeptide).
• Various specific and non-specific cross-linking chemistries are known in the art, such as Click chemistry and other methods. In general, any cross-linking chemistry used to link two proteins may be adapted for use in the presently disclosed VLPs.
• chemistries used in creation of immunoconjugates or antibody drug conjugates may be used.
• an VLP is created using a cleavable or non-cleavable linker.
• the components of the VLP of the present disclosure may have any of various amino acids sequences.
• U.S. Patent Pub No. US 2015/0356240 Al describes various methods for designing protein assemblies.
• the polypeptides were designed for their ability to self-assemble in pairs to form VLPs, such as icosahedral particles.
• the design involved design of suitable interface residues for each member of the polypeptide pair that can be assembled to form the VLP.
• the VLPs so formed include symmetrically repeated, non-natural, non-covalent polypeptide-polypeptide interfaces that orient a first assembly and a second assembly into a VLP, such as one with an icosahedral symmetry.
• Non-limiting examples of designed protein complexes useful in protein-based VLPs of the present disclosure include those disclosed in U.S. Patent No. 9,630,994; IntT Pat. Pub No. WO2018187325A1; U.S. Pat. Pub. No. 2018/0137234 Al; U.S. Pat. Pub. No. 2019/0155988 A2, each of which is incorporated herein in its entirety.
• Illustrative sequences are provided in Table 1. Table 1
• the first multimerization domain comprises an amino acid sequence which is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence of any one of SEQ ID NOs: 24 and 30-31.
• the second multimerization domain comprises an amino acid sequence which is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence of SEQ ID NO: 22-23 and 25-26.
• the VLP comprises a fusion protein that has at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to any one of SEQ ID NO: 1-10 and comprises an RSV F protein as disclosed herein; and a second component that has at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to any one of SEQ ID NO: 22-23, 25-29, and 32.
• the VLP comprises a fusion protein that has at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 6 and comprises an RSV F protein as disclosed herein; and a second component that has at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NO: 26.
• the first component comprises the polypeptide sequence that has at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to any one of SEQ ID NOs: 14, 34, and 35.
• the first component may comprise an RSV F protein, which may be a full-length RSV F protein, the ectodomain of RSV F, or an antigenic fragment thereof.
• the RSV F protein comprises the polypeptide sequence that has at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to any one of SEQ ID NOs: 14, 34, and 35.
• the RSV F protein is an RVS-A F protein.
• the RSV F protein is an RSV-B F protein.
• the first component comprises the polypeptide sequence that has at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to SEQ ID NOs: 14 and further comprises a signal peptide.
• the signal peptide comprises the sequence of any one of SEQ ID NOs: 11-13.
• polypeptides as described herein may have one of more amino acid substitutions relative to wildtype RSV.
• the second component of a protein complex provided herein may comprise 1, 2, 3, 4, 5, 6, 7, or all 8 positions relative to SEQ ID NO: 14 selected from any one of H9Y, E24D, A28Q, A36E, R38A, N97D, N105S, D121H,
• a polypeptide provided herein may comprise one or more conservative amino acid substitutions.
• conservative amino acid substitution is well known in the art and relates to substitution of a particular amino acid by one having a similar characteristic (e.g, similar charge or hydrophobicity).
• Conservative mutations can include, without limitation, substitution of amino acid residues with e.g, similar charge or hydrophobicity but differing in size or bulkiness (e.g., to provide a cavity-filling function).
• a list of conservative amino acid substitutions is given in the table below.
• Non-conservative amino acid substitution may be preferred, for example, for eradication of a flexible portion of the native RSV F protein secondary structure is desired, for example, by adding a cysteine residue (or vice versa).
• “Nonconservative substitution” refers to the substitution of an amino acid in one class with an amino acid from another class; for example, substitution of an Ala with Asp, Asn, Glu, or Gin.
• non-conservative substitutions include the substitution of a non-polar (hydrophobic) amino acid residue such as isoleucine, valine, leucine, alanine, methionine for a polar (hydrophilic) residue such as cysteine, glutamine, glutamic acid or lysine and/or a polar residue for a non-polar residue.
• a non-polar (hydrophobic) amino acid residue such as isoleucine, valine, leucine, alanine, methionine for a polar (hydrophilic) residue such as cysteine, glutamine, glutamic acid or lysine and/or a polar residue for a non-polar residue.
• D- Cys for D-Ala, D-Ser, or D-Tyr may be used to remove intramolecular disulfide bonds, which may, in some cases improve protein stability or expression.
• substitutions to D-Cys may be used to generated disulfide bonds that stability a protein
• the disclosure provides nucleic acids encoding a polypeptide or fusion protein of the disclosure.
• the nucleic acid sequence may comprise RNA (such as mRNA) or DNA.
• Such nucleic acid sequences may comprise additional sequences useful for promoting expression and/or purification of the encoded protein, including but not limited to polyA sequences, modified Kozak sequences, and sequences encoding epitope tags, export signals, and secretory signals, nuclear localization signals, and plasma membrane localization signals. It will be apparent to those of skill in the art, based on the teachings herein, what nucleic acid sequences will encode the proteins of the invention.
• disclosure provides expression vectors comprising the isolated nucleic acid of any embodiment or combination of embodiments of the disclosure operatively linked to a suitable control sequence.
• “Expression vector” includes vectors that operatively link a nucleic acid coding region or gene to any control sequences capable of effecting expression of the gene product.
• “Control sequences” operably linked to the nucleic acid sequences of the disclosure are nucleic acid sequences capable of effecting the expression of the nucleic acid molecules. The control sequences need not be contiguous with the nucleic acid sequences, so long as they function to direct the expression thereof.
• intervening untranslated yet transcribed sequences can be present between a promoter sequence and the nucleic acid sequences and the promoter sequence can still be considered “operably linked” to the coding sequence.
• Other such control sequences include, but are not limited to, polyadenylation signals, termination signals, and ribosome binding sites.
• Such expression vectors can be of any type known in the art, including but not limited to plasmid and viral-based expression vectors.
• control sequence used to drive expression of the disclosed nucleic acid sequences in a mammalian system may be constitutive (driven by any of a variety of promoters, including but not limited to, CMV, SV40, RSV, actin, EF) or inducible (driven by any of a number of inducible promoters including, but not limited to, tetracycline, ecdysone, steroidresponsive).
• the present disclosure provides cells comprising the polypeptide, the virus-like particle, the composition, the nucleic acid, and/or the expression vector of any embodiment or combination of embodiments of the disclosure, wherein the cells can be either prokaryotic or eukaryotic, such as mammalian cells.
• the cells may be transiently or stably transfected with the nucleic acids or expression vectors of the disclosure.
• transfection of expression vectors into prokaryotic and eukaryotic cells can be accomplished via any technique known in the art.
• a method of producing a polypeptide according to the invention is an additional part of the invention. The method comprises the steps of (a) culturing a host according to this aspect of the invention under conditions conducive to the expression of the polypeptide, and (b) optionally, recovering the expressed polypeptide.
• compositions/vaccines comprising
• the virus-like particles elicit potent and protective antibody responses against RSV (e.g., against RSV-A and/or RSV-B).
• RSV e.g., against RSV-A and/or RSV-B
• the virus-like particles of the disclosure induce neutralizing antibody titers.
• a pharmaceutical composition or vaccine provided herein may be bivalent, e.g., may comprise both RSV-A and RSV-B antigens (e.g., both RSV-A and RSV-B F proteins).
• compositions/vaccines may further comprise (a) a lyoprotectant; (b) a surfactant; (c) a bulking agent; (d) a tonicity adjusting agent; (e) a stabilizer; (I) a preservative and/or (g) a buffer.
• the buffer in the pharmaceutical composition is a Tris buffer, a histidine buffer, a phosphate buffer, a citrate buffer or an acetate buffer.
• the composition may also include a lyoprotectant, e.g. sucrose, sorbitol or trehalose.
• the composition includes a preservative e.g.
• the composition includes a bulking agent, like glycine.
• the composition includes a surfactant e.g, polysorbate-20, polysorbate-40, polysorbate- 60, polysorbate-65, polysorbate-80 polysorbate-85, poloxamer-188, sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, sorbitan monooleate, sorbitan trilaurate, sorbitan tristearate, sorbitan trioleaste, or a combination thereof.
• the composition may also include a tonicity adjusting agent, e.g, a compound that renders the formulation substantially isotonic or isoosmotic with human blood.
• Illustrative tonicity adjusting agents include sucrose, sorbitol, glycine, methionine, mannitol, dextrose, inositol, sodium chloride, arginine and arginine hydrochloride.
• the composition additionally includes a stabilizer, e.g, a molecule which substantially prevents or reduces chemical and/or physical instability of the nanostructure, in lyophilized or liquid form.
• Illustrative stabilizers include sucrose, sorbitol, glycine, inositol, sodium chloride, methionine, arginine, and arginine hydrochloride.
• the virus-like particles may be the sole active agent in the composition, e.g., formulated as an aqueous vaccine, or the composition may further comprise one or more other agents suitable for an intended use, including but not limited to adjuvants to stimulate the immune system generally and improve immune responses overall. Any suitable adjuvant can be used.
• adjuvant refers to a compound or mixture that enhances the immune response to an antigen.
• Illustrative types of adjuvants that may be used in a pharmaceutical composition provided herein include the following: 1. mineral-containing compositions; 2. oil emulsions; 3. saponin formulations; 4. virosomes and virus-like particles; 5. bacterial or microbial derivatives; 6. bioadhesives and mucoadhesives; 7. liposomes; 8. polyoxyethylene ether and polyoxyethylene ester formulations; 9. polyphosphazene (pcpp); 10. muramyl peptides; 11. imidazoquinolone compounds; 12. thiosemicarbazone compounds; 13. tryptanthrin compounds; 14. human immunomodulators; 15. lipopeptides; 16. benzonaphthyri dines; 17. microparticles; 18. immunostimulatory polynucleotide (such as RNA or DNA; e.g, c ⁇ g- containing oligonucleotides).
• immunostimulatory polynucleotide such as RNA or DNA;
• Illustrative adjuvants that may be used in a pharmaceutical composition provided herein include, but are not limited to, 3M-052, Adju-PhosTM, AlhydrogelTM, AdjumerTM, albumin-heparin microparticles, Algal Glucan, Algammulin, Alum, Antigen Formulation, AS-2 adjuvant, ASO1, ASO3, autologous dendritic cells, autologous PBMC, AvridineTM, B7-2, BAK, BAY R1005, BECC TLR-4 agonists, Bupivacaine, Bupivacaine-HCl, BWZL, Calcitriol, Calcium Phosphate Gel, CCR5 peptides, CFA, Cholera holotoxin (CT) and Cholera toxin B subunit (CTB), Cholera toxin Al -subunit-Protein A D-fragment fusion protein, CpG, CPG-1018, CRL1005, Cytokine-containing Liposomes
• an adjuvant depends on the subject to be treated.
• a pharmaceutically acceptable adjuvant is used.
• the adjuvant is an aluminum hydroxide gel (e.g, AlhydrogelTM).
• the adjuvant is SWE.
• the adjuvant is MF59.
• the adjuvant is an oil-in-water emulsion.
• the composition may include an aluminum salt adjuvant, an oil in water emulsion (e.g. an oil-in-water emulsion comprising squalene, such as MF59, SWE, or AS03), a TLR9 agonist (such as CpG oligodeoxynucleotides), a TLR7 agonist (such as imidazoquinoline or imiquimod), or a combination thereof.
• the adjuvant is a combination of an aluminum salt and CPG1018.
• Suitable aluminum salts include hydroxides (e.g. oxyhydroxides), phosphates (e.g. hydroxyphosphates, orthophosphates), (e.g.
• the salts can take any suitable form (e.g. gel, crystalline, amorphous, etc.), with adsorption of antigen to the salt being an example.
• the concentration of Al +++ in a composition for administration to a patient may be less than 5mg/ml e.g. ⁇ 4 mg/ml, ⁇ 3 mg/ml, ⁇ 2 mg/ml, ⁇ 1 mg/ml, etc. An illustrative range is between 0.3 and 1 mg/ml.
• Aluminum hydroxide and aluminum phosphate adjuvants are suitable for use with the disclosure.
• a pharmaceutical composition provided herein comprises aluminum hydroxide as an adjuvant.
• a pharmaceutical composition provided herein comprises 500 ⁇ g aluminum hydroxide.
• the composition including the virus-like particles may be the sole active agent in the composition, where no adjuvant is included, or wherein the composition is substantially free of an adjuvant, including substantially free of any adjuvant.
• no adjuvant may be added, or substance(s) having adjuvant property present but minimal quantities, such as quantities not expected to exert an adjuvant effect.
• the pharmaceutical composition has less than about 5%, less than about 4%, less than about 4%, less than about 3%, less than about 2%, less than about 1%, less than about 0.5%, less than about 0.1%, less than 5%, less than 4%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, or less than 0.1% (w/v) of the adjuvant.
• the composition including the virus-like particles may be the sole active agent in the composition and is free of an adjuvant, (e.g,, Alum or aluminum salt adjuvants).
• the unit dose comprises about 1 ⁇ g to about 5 ⁇ g, about 5 ⁇ g to about 10 ⁇ g, about 10 ⁇ g to about 15 ⁇ g, about 15 ⁇ g to about 20 ⁇ g, about 20 ⁇ g to about 30 ⁇ g, about 30 ⁇ g to about 40 ⁇ g, about 40 ⁇ g to about 50 ⁇ g, about 50 ⁇ g to about 60 ⁇ g, about 60 ⁇ g to about 70 ⁇ g, about 70 ⁇ g to about 80 ⁇ g, about 80 ⁇ g to about 90 ⁇ g, about 90 ⁇ g to about 100 ⁇ g, about 100 ⁇ g to about 110 ⁇ g, about 110 ⁇ g to about 120 ⁇ g, about 120 ⁇ g to about 130 ⁇ g, about 130 ⁇ g to about 140 ⁇ g, about 140 ⁇ g to about 150 ⁇ g, about 150 ⁇ g to about 200 ⁇ g, about 200 ⁇ g to about 250 ⁇
• the unit dose comprises about 1 ⁇ g, about 2 ⁇ g, about 5 ⁇ g, about 10 ⁇ g, about 15 ⁇ g, about 25 ⁇ g , about 50 ⁇ g, about 75 ⁇ g, about 100 ⁇ g, about 125 ⁇ g, about 150 ⁇ g, about 200 ⁇ g, or about 250 ⁇ g of the protein complex.
• the unit dose comprises, 25 ⁇ g, 75 ⁇ g, or 250 ⁇ g of the protein complex.
• the abbreviation ⁇ g may be used interchangeably with the abbreviation mcg to refer to micrograms of a substance.
• the unit dosage comprises 5 ⁇ g of the protein complex.
• the unit dosage comprises 25 ⁇ g of the protein complex. In some embodiments, the unit dosage comprises 125 ⁇ g of the protein complex. In some embodiments, the unit dosage comprises 100 ⁇ g of the protein complex. In another aspect, provided herein is a unit dose of the pharmaceutical composition described herein, wherein the unit dose comprises 2 ⁇ g, 5 ⁇ g, 10 ⁇ g, 15 ⁇ g, 25 ⁇ g, 50 ⁇ g, 75 ⁇ g, 100 ⁇ g, or 125 ⁇ g of the protein complex. In some embodiments, provided herein is a unit dose of the pharmaceutical composition described herein, wherein the unit dose comprises between about 25 ⁇ g and about 125 ⁇ g of the protein complex.
• the unit dose of the pharmaceutical composition is between about 2 ⁇ g to about 125 ⁇ g, or between about 5 ⁇ g to about 125 g, or between about 15 ⁇ g to 125 ⁇ g, or between about 25 ⁇ g to about 125 ⁇ g, or between about 50 ⁇ g to about 125 ⁇ g, or between about 100 ⁇ g to about 125 ⁇ g of the protein complex.
• a unit dose of the pharmaceutical composition described herein wherein the unit dose comprises between about 25 ⁇ g and about 125 ⁇ g of the protein complex.
• the unit dose of the pharmaceutical composition is between about 2 ⁇ g to about 125 ⁇ g, or between about 5 ⁇ g to about 125 g, or between about 15 ⁇ g to 125 ⁇ g, or between about 25 ⁇ g to about 125 ⁇ g, or between about 50 ⁇ g to about 125 ⁇ g, or between about 100 ⁇ g to about 125 ⁇ g of the protein complex.
• about 10 ⁇ g to about 100 ⁇ g, about 10 ⁇ g to about 150 ⁇ g, about 10 ⁇ g to about 200 ⁇ g, about 10 ⁇ g to about 250 ⁇ g, about 10 ⁇ g to about 300 ⁇ g, about 10 ⁇ g to about 350 ⁇ g, about 10 ⁇ g to about 400 ⁇ g, about 10 ⁇ g to about 450 ⁇ g, or about 10 ⁇ g to about 500 ⁇ g of the protein complex are administered.
• about 25 ⁇ g to about 100 ⁇ g, about 25 ⁇ g to about 150 ⁇ g, about 25 ⁇ g to about 200 ⁇ g, about 25 ⁇ g to about 250 ⁇ g, about 25 ⁇ g to about 300 ⁇ g, about 25 ⁇ g to about 350 ⁇ g, about 25 ⁇ g to about 400 ⁇ g, about 25 ⁇ g to about 450 ⁇ g, or about 25 ⁇ g to about 500 ⁇ g of the protein complex are administered.
• about 50 ⁇ g to about 100 ⁇ g, about 50 ⁇ g to about 150 ⁇ g, about 50 ⁇ g to about 200 ⁇ g, about 50 ⁇ g to about 250 ⁇ g, about 50 ⁇ g to about 300 ⁇ g, about 50 ⁇ g to about 350 ⁇ g, about 50 ⁇ g to about 400 ⁇ g, about 50 ⁇ g to about 450 ⁇ g, or about 50 ⁇ g to about 500 ⁇ g of the protein complex are administered.
• about 5 ⁇ g to about 150 ⁇ g, about 10 ⁇ g to about 150 ⁇ g, about 25 ⁇ g to about 150 ⁇ g, about 50 ⁇ g to about 150 ⁇ g, about 75 ⁇ g to about 150 ⁇ g, about 100 ⁇ g to about 150 ⁇ g, or about 125 ⁇ g to about 150 ⁇ g of the protein complex are administered.
• about 5 ⁇ g to about 125 ⁇ g, about 10 ⁇ g to about 125 ⁇ g, about 25 ⁇ g to about 125 ⁇ g, about 50 ⁇ g to about 125 ⁇ g, about 75 ⁇ g to about 125 ⁇ g, or about 100 ⁇ g to about 125 ⁇ g of the protein complex are administered.
• about 5 ⁇ g to about 100 ⁇ g, about 10 ⁇ g to about 100 ⁇ g, about 25 ⁇ g to about 100 ⁇ g, about 50 ⁇ g to about 100 ⁇ g, or about 75 ⁇ g to about 100 ⁇ g of the protein complex are administered.
• about 5 ⁇ g to about 75 ⁇ g, about 10 ⁇ g to about 75 ⁇ g, about 25 ⁇ g to about 75 ⁇ g, or about 50 ⁇ g to about 75 ⁇ g of the protein complex are administered.
• about 5 ⁇ g to about 50 ⁇ g, about 10 ⁇ g to about 50 ⁇ g, or about 25 ⁇ g to about 50 ⁇ g of the protein complex are administered.
• the protein complexes of the disclosure comprise an RSV F protein.
• the total molecular mass of the protein complex is about 5.6 MDa and the mass fraction of the RSV F protein in the protein complex is about 58%; there are 20 copies of the RSV F protein trimer per protein complex in these embodiments.
• the dose amounts described above may be converted to a molar amount, where 1 ⁇ g equals about 0.18 picomoles (pmol) of the protein complex or about 3.6 pmol of RSV F protein within the protein complex. Each 1 ⁇ g of protein complex equals about 0.6 ⁇ g of RSV F protein.
• a unit dose of 25 ⁇ g of protein complex is equal to a unit dose of about 14 ⁇ g of a non-particle-associated RSV F protein trimer; conversely, a unit dose of about 100 ⁇ g of a non-particle-associated RSF F protein trimer is equivalent to about 174 ⁇ g of such an illustrative protein complex.
• the pH of the formulation can also vary. In general, it is between about pH 6.2 to about pH 8.0. In some embodiments, the pH is about 6.2, about 6.4, about 6.6, about 6.8, about 7.0, about 7.2, about 7.4, about 7.6, about 7.8, or about 8.0. Of course, the pH may also be within a range of values. Thus, in some embodiments the pH is between about 6.2 and about 8.0, between about 6.2 and 7.8, between about 6.2 and 7.6, between about 6.2 and 7.4, between about 6.2 and 7.2, between about 6.2 and 7.0, between about 6.2 and 6.8, between about 6.2 and about 6.6, or between about 6.2 and 6.4.
• the pH is between 6.4 and about 8.0, between about 6.4 and 7.8, between about 6.4 and 7.6, between about 6.4 and 7.4, between about 6.4 and 7.2, between about 6.4 and 7.0, between about 6.4 and 6.8, or between about 6.4 and about 6.6.
• the pH is between about 6.6 and about 8.0, between about 6.6 and 7.8, between about 6.6 and 7.6, between about 6.6 and 7.4, between about 6.6 and 7.2, between about 6.6 and 7.0, or between about 6.6 and 6.8.
• it is between about 6.8 and about 8.0, between about 6.8 and 7.8, between about 6.8 and 7.6, between about 6.8 and 7.4, between about 6.8 and 7.2, or between about 6.8 and 7.0.
• it is between about 7.0 and about 8.0, between about 7.0 and 7.8, between about 7.0 and 7.6, between about 7.0 and 7.4, between about 7.0 and 7.2, between about 7.2 and 8.0, between about 7.2 and 7.8, between about 7.2 and about 7.6, between about 7.2 and 7.4, between about 7.4 and about 8.0, about 7.4 and about 7.6, or between about 7.6 and about 8.0.
• the formulation can include one or more salts, such as sodium chloride, sodium phosphate, or a combination thereof.
• each salt is present in the formulation at about 10 mM to about 200 mM.
• any salt that is present is present at about 10 mM to about 200 mM, about 20 mM to about 200 mM, about 25 mM to about 200 mM, at about 30 mM to about 200 mM, at about 40 mM to about 200 mM, at about 50 mM to about 200 mM, at about 75 mM to about 200 mM, at about 100 mM to about 200 mM, at about 125 mM to about 200 mM, at about 150 mM to about 200 mM, or at about 175 mM to about 200 mM.
• any salt that is present is present at about 10 mM to about 175 mM, about 20 mM to about 175 mM, about 25 mM to about 175 mM, at about 30 mM to about 175 mM, at about 40 mM to about 175 mM, at about 50 mM to about 175 mM, at about 75 mM to about 175 mM, at about 100 mM to about 175 mM, at about 125 mM to about 175 mM, or at about 150 mM to about 175 mM.
• any salt that is present is present at about 10 mM to about 150 mM, about 20 mM to about 150 mM, about 25 mM to about 150 mM, at about 30 mM to about 150 mM, at about 40 mM to about 150 mM, at about 50 mM to about 150 mM, at about 75 mM to about 150 mM, at about 100 mM to about 150 mM, or at about 125 mM to about 150 mM.
• any salt that is present is present at about 10 mM to about 125 mM, about 20 mM to about 125 mM, about 25 mM to about 125 mM, at about 30 mM to about 125 mM, at about 40 mM to about 125 mM, at about 50 mM to about 125 mM, at about 75 mM to about 125 mM, or at about 100 mM to about 125 mM.
• any salt that is present is present at about 10 mM to about 100 mM, about 20 mM to about 100 mM, about 25 mM to about 100 mM, at about 30 mM to about 100 mM, at about 40 mM to about 100 mM, at about 50 mM to about 100 mM, or at about 75 mM to about 100 mM.
• any salt that is present is present at about 10 mM to about 75 mM, about 20 mM to about 75 mM, about 25 mM to about 75 mM, at about 30 mM to about 75 mM, at about 40 mM to about 75 mM, or at about 50 mM to about 75 mM.
• any salt that is present is present at about 10 mM to about 50 mM, about 20 mM to about 50 mM, about 25 mM to about 50 mM, at about 30 mM to about 50 mM, or at about 40 mM to about 50 mM.
• any salt that is present is present at about 10 mM to about 40 mM, about 20 mM to about 40 mM, about 25 mM to about 40 mM, at about 30 mM to about 40 mM, at about 10 mM to about 30 mM, at about 20 mM to about 30, at about 25 mM to about 30 mM, at about 10 mM to about 25 mM, at about 20 mM to about 25 mM, or at about 10 mM to about 20 mM.
• the sodium chloride is present in the formulation at about 100 mM.
• the sodium phosphate is present in the formulation at about 25 mM.
• Formulations herein may further comprise a solubilizing agent such as a nonionic detergent.
• a solubilizing agent such as a nonionic detergent.
• Such detergents include, but are not limited to polysorbate 80 (Tween® 80), TritonXIOO and polysorbate 20.
• a pharmaceutical composition described herein may comprise a polypeptide, a virus-like particle, composition, a nucleic acid, an expression vector, and/or a cell of embodiment or combination of embodiments herein and one or more additional vaccines, as well as a pharmaceutically acceptable carrier.
• the one or more additional vaccines is a pediatric vaccine.
• Vaccines that may be coformulated with a the polypeptide, the virus-like particle, the composition, the nucleic acid, the expression vector, and/or the cell of embodiment or combination described herein include, without limitation, a vaccine against Hepatitis B, a vaccine against Rotavirus, a vaccines against diphtheria, tetanus and pertussis (“DTaP”), a vaccine against polio, a vaccine against influenza, and a vaccine against measles, mumps and rubella (“MMR”).
• a vaccine against Hepatitis B a vaccine against Rotavirus
• DTaP diphtheria, tetanus and pertussis
• MMR measles
• MMR measles, mumps and rubella
• the disclosure provides methods to vaccinate a subject against infection with RSV (e.g., infection with RSV-A and/or RSV-B), comprising administering to a subject in need thereof an amount effective to treat or limit development of the infection of the polypeptide, virus-like particle, composition, nucleic acid, pharmaceutical composition, or vaccine of any embodiment herein (referred to as the “immunogenic composition”).
• RSV infection with RSV-A and/or RSV-B
• administering comprising administering to a subject in need thereof an amount effective to treat or limit development of the infection of the polypeptide, virus-like particle, composition, nucleic acid, pharmaceutical composition, or vaccine of any embodiment herein (referred to as the “immunogenic composition”).
• the method prevents disease following infection with RSV subtypes A and/or B.
• such a method protects against the development of RSV-associated disease (e.g., severe disease), for example, pneumonia and/or acute respiratory disease.
• the subject may be any suitable mammalian subject, including but not limited to a human subject.
• a subject is a human child, e.g., a child of less than 12 months of age.
• a subject is a human toddler, e.g., of about 1 to about 3 years of age or of about 1 to about 5 years of age.
• the subject is a human adult of more than 60 years of age.
• the subject is a human adult of more than 65 years of age.
• the subject is dependent on the help of others or with serious health concerns or risks (e.g. a frail elderly person).
• the subject is a healthy adult of 18-60 years of age.
• the subject is a healthy adult of 18-45 years of age. In another embodiment, the subject is a pregnant woman. In some embodiments the subject is an immunocompromised human adult. In some embodiments, the subject is a human adult suffering from chronic underlying heart and/or lung disease or from functional disability. In some embodiments, the subject is at risk of severe RSV disease (e.g., LRTI or pneumonia).
• severe RSV disease e.g., LRTI or pneumonia
• the immunogenic compositions provided herein may be used vaccinate an unborn child.
• the administration of certain inactivated vaccines are recommended during pregnancy to induce immunity in the unborn child, for example, the tetanus toxoid, reduced diphtheria toxoid and acellular pertussis (Tdap) vaccine and the influenza vaccine.
• Tdap acellular pertussis
• provide herein is a method of generating an immune response in an unborn child, the method comprising administering an effective amount of the immunogenic composition provided herein to the mother of said unborn child.
• the immunogenic composition may be administered at any suitable time point in the pregnancy, e.g., in the last trimester of the pregnancy.
• An immunogenic composition provided herein may be co-administered with other treatments, such as other vaccines.
• a subject treated in accordance with a method provided herein may also be administered one or more seasonal or pandemic vaccines such as an influenza vaccine or a SARS-Cov2 vaccine.
• a subject treated in accordance with the methods provided herein may also be administered a pneumococcal, Recombinant Zoster (Shingles), or Tdap vaccine.
• One, two, or more vaccines may be co-administered with an immunogenic composition provided herein. “Co-administration” includes both concurrently as well as subsequent administration.
• the one, two, or more vaccines and an immunogenic composition provided herein may be administered on the same day.
• the one, two, or more vaccines and an immunogenic composition provided herein are administered within 1 hour, within 2 hours, within 3 hours, within 4 hours, within 5 hours, within 6 hours, within 8 hours, withing 10 hours, or within 12 hours.
• provide herein is a method of treating a subject suffering from RSV infection.
• “treat” or “treating” includes, but is not limited to accomplishing one or more of the following: (a) reducing RSV titer (e.g., RSV-A titer and/or RSV-B titer) in the subject; (b) limiting any increase of RSV titer (e.g., RSV-A titer and/or RSV-B titer) in the subject; (c) reducing the severity of RSV infection symptoms (e.g., RSV- A infection and/or RSV-B infection); (d) limiting or preventing development of symptoms after RSV infection (e.g., RSV-A infection and/or RSV-B infection); (e) inhibiting worsening of symptoms of RSV infection (e.g., RSV-A infection and/or RSV-B infection); (f) limiting or preventing recurrence of symptoms of RSV infection (e.g., RSV-A infection and/or RSV-B infection)
• a method of vaccinating decreases the subject’s risk of becoming infected with RSV (e.g., RSV-A and/or RSV-B). In some embodiments, a method of vaccinating limits the development of an RSV infection (e.g., RSV-A infection and/or RSV-B infection). In some embodiments, a method of vaccinating decreases the severity of the symptoms of an RSV infection (e.g., RSV-A infection and/or RSV-B infection). In some embodiments, the infection with RSV (e.g., infection with RSV-A and/or RSV-B) is a lower respiratory tract infection (LRTI).
• LRTI lower respiratory tract infection
• the methods provided herein may be used to prevent an RSV infection or illness (e.g., pneumonia or acute respiratory disease) in a subject.
• “prevent” or “preventing” includes, but is not limited to accomplishing one or more of the following: (a) generating an immune response (antibody and/or cell-based, e.g., CD4 T cells, memory B cells, and/or CD8 T cells) to RSV (e.g., RSV-A and/or RSV-B) in the subject expected to confer protection against lower respiratory tract infection (LRTI) caused by or associated with RSV in the subject; (b) generating neutralizing antibodies against RSV (e.g., RSV-A and/or RSV-B) in the subject expected to reduce the severity of LRTI caused by or associated with RSV in the subject; (c) preventing LRTI caused by or associated with RSV in a subject, detected as an increase in the titer of the virus i of the subject or by an increase in
• RSV-A infection and/or RSV-B infection refers to accomplishing one or more of the following: (a) generating an immune response (antibody and/or cell-based, e.g., CD4 T cells, memory B cells, and/or CD8 T cells) to RSV (e.g., RSV-A and/or RSV-B) in the subject expected to limit an increase in viral titer or symptoms in the subject; (b) generating neutralizing antibodies against RSV (e.g., RSV-A and/or RSV-B) in the subject at a level expected to limit an increase in viral titer or symptoms in the subject; (c) causing reduced RSV titers (e.g., RSV-A and/or RSV-B titers) in the subject after exposure to RSV (e.g., RSV-A and/or RSV-B) compared to subjects not administered the protein complex;
• an immune response antibody and/or cell-based, e.g., CD4 T cells,
• the methods provided herein may be used to prevent or limit development of infection with an RSV-A subtype and/or an RSV-B subtype.
• the methods provided herein may be used to prevent or limit development of infection with an original strain of RSV and/or infection with a variant strain of RSV.
• variant RSV strains include, without limitation, RSV ONI, RSV NA1, RSV LBA1, RSV LBA2, RSV BA, RSV Long, RAV A2, and others (see, e.g., Pandya et al., Pathogens 2019, 8(2), 67; and Melero and Moore, Curr Top Microbiol Immunol. 2013; 372: 59-82).
• the pharmaceutical composition of the present invention may be effective in preventing or limiting infection of RSV strains that have not yet been described or discovered.
• Clinical efficacy of a respiratory virus vaccine can be assessed by various means known in the art, including but not limited to placebo-controlled clinical efficacy studies to measure viral load or symptoms of RSV disease in vaccinated versus control subjects. Correlates of protection may also be defined, such as neutralizing antibody titers (typically expressed as geometric mean titers), fold increases above baselines (typically expressed as geometric fold rise), and seroresponse rate (a percentage of subjects that achieve a fold rise in neutralizing antibody titers above a predetermined threshold).
• Guidance on direct and surrogate measures of clinical efficacy for respiratory diseases are available, for example, in Guidance for Industry: Clinical Data Needed to Support the Licensure of Seasonal Inactivated Influenza Vaccines. U.S. Food & Drug Administration (May 2007) and Respiratory Syncytial Virus Infection: Developing Antiviral Drugs for Prophylaxis and Treatment Guidance for Industry. U.S. Food & Drug Administration (October 2017).
• the methods described herein generate an immune response in a subject in the subject not known to be infected with RSV (e.g., RSV-A and/or RSV-B), wherein the immune response serves to limit development of infection and symptoms of an RSV (e.g., RSV-A and/or RSV-B) infection.
• the immune response comprises generation of neutralizing antibodies and/or cell-based responses against RSV (e.g., RSV-A and/or RSV-B).
• the immune response comprises generation of RSV F protein-specific (e.g., RSV-A and/or RSV-B F protein-specific) responses with a mean geometric titer of at least 1 x 10 3 , at least 1 x 10 4 , at least 1 x 10 5 , at least 1 x 10 6 , at least 1 x 10 7 , at least 1 x 10 8 , or at least 1 x 10 9 .
• the immune response comprises generation of antibodies against multiple antigenic epitopes or RSV (e.g., RSV-A and/or RSV-B).
• the methods provided herein may results in an increase in antibody titers in a subject, e.g., in an increase in RSV-A-specific neutralizing antibodies, RSV-B- specific neutralizing antibodies, RSV F-protein-specific IgG antibodies, and/or RSV F- protein-specific neutralizing antibodies.
• Antibody titers may be determined using any suitable assays known in the art or described herein including, without limitation, binding enzyme-linked immunosorbent assays (ELISA), enzyme-linked immune absorbent spot (ELISpot), competition ELISAs, immunoprecipitation, immunoblotting, and agglutination assays.
• a neutralization or microneutralization (MN) assay may be used to measure increases in neutralizing antibodies in a subject after administration of a protein complex as described herein.
• Microneutralization refers to a neutralizing performed in a miniaturized format, such as a 96-well plate.
• a (micro)neutralization assay is used to test for the inhibition of a virus by antibodies (e.g., purified antibodies, serum, or plasma). The assay measures the level of antibodies present in a sample that are able to neutralize a virus in vitro.
• microneutralization assays for clinical samples are performed with a serial dilution of serum mixed with a fixed concentration of virus.
• microneutralization assays Methods for performing (micro)neutralization assays are well known. Illustrative microneutralization assays are described, see, e.g., van Baalen et al. Vaccine 35 (2017) 46-52. [0101] If neutralizing antibodies specific for RSV are present in a sample, the virus will be neutralized, and infection of cells (e.g., HEp-2 cells) is inhibited. Immunofluorescence levels indicating viral infection may analyzed using, e.g., a CTL ImmunoSpot® UV analyzer, equipped with BioSpot® analysis software for automated counting of infected cells. Results are generally reported in international units per milliliter (lU/mL). Validation and standardized of microneutralization assays is described in the Examples below.
• the methods provided herein result in an increase in antibodies (e.g., RSV-A-specific neutralizing antibodies, RSV-B-specific neutralizing antibodies, RSV F-protein-specific IgG antibodies, RSV F-protein-specific neutralizing antibodies, and/or antibodies against human metapneumo virus) of about 1-fold to about 3- fold, about 3-fold to about 4-fold, about 4-fold to about 5-fold, about 5-fold to about 6-fold, about 6-fold to about 7-fold, about 7-fold to about 8-fold, about 8-fold to about 9-fold, about 9-fold to about 10-fold, about 10-fold to about 12-fold, about 12-fold to about 15-fold, about 15-fold to about 20-fold, about 20-fold to about 25-fold, about 25-fold to about 30-fold, about 30-fold to about 40-fold, about 40-fold to about 50-fold, about 50-fold to about 60- fold, about 60-fold to about 70-fold, about 70-fold to about 80-fold, about 80-fold to about 90
• antibodies e.g.
• the methods provided herein result in an increase in antibodies (e.g., in an increase in RSV-A-specific neutralizing antibodies, RSV-B-specific neutralizing antibodies, RSV F-protein-specific IgG antibodies, RSV F-protein-specific neutralizing antibodies, and/or antibodies against human metapneumo virus) of at least about 3 -fold, at least about 4-fold, at least about 5 -fold, at least about 10-fold, at least about 15 -fold, at least about 20-fold, or at least about 25-fold compared to baseline.
• antibodies e.g., in an increase in RSV-A-specific neutralizing antibodies, RSV-B-specific neutralizing antibodies, RSV F-protein-specific IgG antibodies, RSV F-protein-specific neutralizing antibodies, and/or antibodies against human metapneumo virus
• baseline is meant a measurement of antibodies immediately prior to administration of the first dose of an immunogenic composition provided herein.
• the increase in antibodies (e.g., RSV-A-specific neutralizing antibodies, RSV-B-specific neutralizing antibodies, RSV F-protein-specific IgG antibodies, RSV F- protein-specific neutralizing antibodies, and/or antibodies against human metapneumovirus) compared to baseline is detectable within about 3 days to about 7 days, about 1 week to about 2 weeks, about 2 weeks to about 3 weeks, about 3 weeks to about 4 weeks, about 4 weeks to about 5 weeks, about 5 weeks to about 6 weeks, about 6 weeks to about 7 weeks, about 7 weeks to about 8 weeks, about 8 weeks to about 9 weeks, about 9 weeks to about 10 weeks, about 10 weeks to about 11 weeks, about 11 weeks to about 12 weeks, about 3 months to about 4 months, about 4 months to about 5 months, about 5 months to about 6 months, about 6 months to about 9 months, about 9 months to about 12 months, about 12 months to about 18 months
• the increase in antibodies compared to baseline is detectable within about 2 weeks, within about 3 weeks, within about 4 weeks, within about 5 weeks, within about 6 weeks, within about 7 weeks, within about 8 weeks, within about 9 weeks, within about 10 weeks, within about 11 weeks, or within about 12 weeks of administration of the immunogenic composition.
• the methods provided herein may result in an increase in immune cells in a subject, e.g., an increase in RSV F-protein-specific memory B cells and/or RSV F-protein-specific T cells.
• the memory B cells and/or T cells may be specific to RSV-A F protein or RSV-B F protein, or they may be reactive to both.
• the number of immune cells in a subject may be determined using any suitable assay known in the art or described herein, including, without limitation, FACS and flow cytometry.
• the methods provided herein results in an increase in immune cells (e.g., in an increase in RSV F-protein-specific memory B cells and/or RSV F-protein-specific T cells) of about 1-fold to about 3-fold, about 3-fold to about 4-fold, about 4-fold to about 5-fold, about 5-fold to about 6-fold, about 6-fold to about 7-fold, about 7-fold to about 8-fold, about 8-fold to about 9-fold, about 9-fold to about 10-fold, about 10-fold to about 12-fold, about 12-fold to about 15-fold, about 15-fold to about 20-fold, about 20-fold to about 25-fold, about 25-fold to about 30-fold, about 30-fold to about 40-fold, about 40- fold to about 50-fold, about 50-fold to about 60-fold, about 60-fold to about 70-fold, about 70-fold to about 80-fold, about 80-fold to about 90-fold, about 90-fold to about 100-fold, or more than about 100-fold compared to baseline.
• immune cells e.g., in
• the methods provided herein result in an increase in immune cells in a subject, e.g., an increase in RSV F-protein-specific memory B cells and/or RSV F-protein-specific T cells.
• the methods provided herein results in an increase in immune cells (e.g., in an increase in RSV F-protein-specific memory B cells and/or RSV F-protein-specific T cells) of at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 10- fold, at least about 15-fold, at least about 20-fold, or at least about 25-fold compared to baseline.
• the memory B cells and/or T cells may be specific to RSV-A F protein or RSV- B F protein, or they may be reactive to both.
• the increase in immune cells compared to baseline is detectable within about 3 days to about 7 days, about 1 week to about 2 weeks, about 2 weeks to about 3 weeks, about 3 weeks to about 4 weeks, about 4 weeks to about 5 weeks, about 5 weeks to about 6 weeks, about 6 weeks to about 7 weeks, about 7 weeks to about 8 weeks, about 8 weeks to about 9 weeks, about 9 weeks to about 10 weeks, about 10 weeks to about 11 weeks, about 11 weeks to about 12 weeks, about 3 months to about 4 months, about 4 months to about 5 months, about 5 months to about 6 months, about 6 months to about 9 months, about 9 months to about 12 months, about 12 months to about 18 months, about 18 months to about 24 months, about 2 years to about 3 years, about 3 years to about 4 years, about 4 years to about 5 years, or about 5 years to about 10 years of administration of the immunogenic composition.
• the increase in increase in immune cells compared to baseline is detectable within about 2 weeks, within about 3 weeks, within about 4 weeks, within about 5 weeks, within about 6 weeks, within about 7 weeks, within about 8 weeks, within about 9 weeks, within about 10 weeks, within about 11 weeks, or within about 12 weeks of administration of the immunogenic composition.
• the memory B cells and/or T cells may be specific to RSV-A F protein or RSV-B F protein, or they may be reactive to both.
• an “effective amount” refers to an amount of the immunogenic composition that is effective for treating and/or limiting RSV infection (e.g., RSV-A infection and/or RSV-B infection).
• the polypeptide, virus-like particle, composition, nucleic acid, pharmaceutical composition, or vaccine of any embodiment herein are typically formulated as a pharmaceutical composition, such as those disclosed above, and can be administered via any suitable route, including intranasally, sublingually, orally, parentally, by inhalation spray, rectally, or topically in dosage unit formulations containing conventional pharmaceutically acceptable carriers, adjuvants, and vehicles.
• parenteral as used herein includes, subcutaneous, intravenous, intra-arterial, intramuscular, intrastemal, intratendinous, intraspinal, intracranial, intrathoracic, infusion techniques or intraperitoneally.
• Polypeptide compositions may also be administered via microspheres, liposomes, immune-stimulating complexes (ISCOMs), or other microparticulate delivery systems or sustained release formulations introduced into suitable tissues (such as blood).
• ISCOMs immune-stimulating complexes
• Dosage regimens can be adjusted to provide the optimum desired response (e.g., a therapeutic or prophylactic response).
• a suitable dosage range may, for instance, be 0.1 ⁇ g/kg to 0.5 ⁇ g /kg body weight, 0.5 ⁇ g/kg to 1 ⁇ g body weight, 1 ⁇ g/kg to 2 ⁇ g/kg body weight, 2 ⁇ g/kg to 3 ⁇ g/kg body weight, 3 ⁇ g/kg to 4 ⁇ g/kg body weight, 4 ⁇ g/kg to 5 ⁇ g/kg body weight, 5 ⁇ g/kg to 6 ⁇ g/kg body weight, 6 ⁇ g/kg to7 ⁇ g/kg body weight, 7 ⁇ g/kg to 8 ⁇ g/kg body weight, 8 ⁇ g/kg to 9 ⁇ g/kg body weight, 9 ⁇ g/kg to 10 ⁇ g/kg body weight, 10 ⁇ g/kg to 15 ⁇ g/kg body weight, 15 ⁇ g/kg to 20 ⁇ g/kg body weight, 20 ⁇ g/kg to 25 ⁇ g/kg body weight, 25 ⁇ g/kg to 30 ⁇ g/kg body weight, 30
• the composition can be delivered in a single bolus, or may be administered more than once (e.g., 2, 3, 4, 5, or more times) as determined by attending medical personnel.
• about 10 ⁇ g to about 100 ⁇ g, about 10 ⁇ g to about 150 ⁇ g, about 10 ⁇ g to about 200 ⁇ g, about 10 ⁇ g to about 250 ⁇ g, about 10 ⁇ g to about 300 ⁇ g, about 10 ⁇ g to about 350 ⁇ g, about 10 ⁇ g to about 400 ⁇ g, about 10 ⁇ g to about 450 ⁇ g, or about 10 ⁇ g to about 500 ⁇ g of the protein complex are administered.
• about 25 ⁇ g to about 100 ⁇ g, about 25 ⁇ g to about 150 ⁇ g, about 25 ⁇ g to about 200 ⁇ g, about 25 ⁇ g to about 250 ⁇ g, about 25 ⁇ g to about 300 ⁇ g, about 25 ⁇ g to about 350 ⁇ g, about 25 ⁇ g to about 400 ⁇ g, about 25 ⁇ g to about 450 ⁇ g, or about 25 ⁇ g to about 500 ⁇ g of the protein complex are administered.
• about 50 ⁇ g to about 100 ⁇ g, about 50 ⁇ g to about 150 ⁇ g, about 50 ⁇ g to about 200 ⁇ g, about 50 ⁇ g to about 250 ⁇ g, about 50 ⁇ g to about 300 ⁇ g, about 50 ⁇ g to about 350 ⁇ g, about 50 ⁇ g to about 400 ⁇ g, about 50 ⁇ g to about 450 ⁇ g, or about 50 ⁇ g to about 500 ⁇ g of the protein complex are administered.
• about 5 ⁇ g to about 150 ⁇ g, about 10 ⁇ g to about 150 ⁇ g, about 25 ⁇ g to about 150 ⁇ g, about 50 ⁇ g to about 150 ⁇ g, about 75 ⁇ g to about 150 ⁇ g, about 100 ⁇ g to about 150 ⁇ g, or about 125 ⁇ g to about 150 ⁇ g of the protein complex are administered.
• about 5 ⁇ g to about 125 ⁇ g, about 10 ⁇ g to about 125 ⁇ g, about 25 ⁇ g to about 125 ⁇ g, about 50 ⁇ g to about 125 ⁇ g, about 75 ⁇ g to about 125 ⁇ g, or about 100 ⁇ g to about 125 ⁇ g of the protein complex are administered.
• about 5 ⁇ g to about 100 ⁇ g, about 10 ⁇ g to about 100 ⁇ g, about 25 ⁇ g to about 100 ⁇ g, about 50 ⁇ g to about 100 ⁇ g, or about 75 ⁇ g to about 100 ⁇ g of the protein complex are administered.
• about 5 ⁇ g to about 75 ⁇ g, about 10 ⁇ g to about 75 ⁇ g, about 25 ⁇ g to about 75 ⁇ g, or about 50 ⁇ g to about 75 ⁇ g of the protein complex are administered.
• about 5 ⁇ g to about 50 ⁇ g, about 10 ⁇ g to about 50 ⁇ g, or about 25 ⁇ g to about 50 ⁇ g of the protein complex are administered.
• about 1 ⁇ g to about 5 ⁇ g, about 5 ⁇ g to about 10 ⁇ g, about 10 ⁇ g to about 15 ⁇ g, or about 15 ⁇ g to about 25 ⁇ g of the protein complex are administered.
• about 1 ⁇ g to about 5 ⁇ g, about 1 ⁇ g to about 10 ⁇ g, about 1 ⁇ g to about 15 ⁇ g, about 1 ⁇ g to about 20 ⁇ g, about 1 ⁇ g to about 25 ⁇ g, about 1 ⁇ g to about 50 ⁇ g, or about 1 ⁇ g to about 75 ⁇ g of the protein complex are administered.
• 1 ⁇ g to 5 ⁇ g, 1 ⁇ g to 10 ⁇ g, 1 ⁇ g to 15 ⁇ g, 1 ⁇ g to 20 ⁇ g, 1 ⁇ g to 25 ⁇ g, 1 ⁇ g to 50 ⁇ g, or 1 ⁇ g to 75 ⁇ g of the protein complex are administered.
• about 5 ⁇ g to about 10 ⁇ g, about 5 ⁇ g to about 15 ⁇ g, about 5 ⁇ g to about 20 ⁇ g, about 5 ⁇ g to about 25 ⁇ g, about 5 ⁇ g to about 50 ⁇ g, or about 5 ⁇ g to about 75 ⁇ g of the protein complex are administered.
• 5 ⁇ g to 10 ⁇ g, 5 ⁇ g to 15 ⁇ g, 5 ⁇ g to 20 ⁇ g, 5 ⁇ g to 25 ⁇ g, 5 ⁇ g to 50 ⁇ g, or 5 ⁇ g to 75 ⁇ g of the protein complex are administered.
• about 10 ⁇ g to about 15 ⁇ g, about 10 ⁇ g to about 20 ⁇ g, about 10 ⁇ g to about 25 ⁇ g, about 10 ⁇ g to about 50 ⁇ g, or about 10 ⁇ g to about 75 ⁇ g of the protein complex are administered.
• 10 ⁇ g to 15 ⁇ g, 10 ⁇ g to 20 ⁇ g, 10 ⁇ g to 25 ⁇ g, 10 ⁇ g to 50 ⁇ g, or 10 ⁇ g to 75 ⁇ g of the protein complex are administered.
• about 25 ⁇ g to about 50 ⁇ g, or about 25 ⁇ g to about 75 ⁇ g of the protein complex are administered.
• 25 ⁇ g to 50 ⁇ g, or 25 ⁇ g to 75 ⁇ g of the protein complex are administered.
• about 50 ⁇ g to about 75 ⁇ g of the protein complex are administered.
• the unit dose of the pharmaceutical composition comprises at most 1 ⁇ g, at most 2.5 ⁇ g, at most 5 ⁇ g, at most 7.5 ⁇ g, at most 10 ⁇ g, at most 12.5 ⁇ g, at most 15 ⁇ g, at most 17.5 ⁇ g, at most 20 ⁇ g, at most 22.5 ⁇ g, or at most 25 ⁇ g of the protein complex, wherein the protein complex comprises 60 copies (20 trimers) of DS- Cavl-I53-50A or DS-Cavl-I53-50AAcys and 60 copies (12 pentamers) of I53-50B, 153- 50B.1, 153-50B. lNegT2 or I53-50B.4PosTl.
• the methods of the disclosure comprise administering at most 1 ⁇ g, at most 2.5 ⁇ g, at most 5 ⁇ g, at most 7.5 ⁇ g, at most 10 ⁇ g, at most 12.5 ⁇ g, at most 15 ⁇ g, at most 17.5 ⁇ g, at most 20 ⁇ g, at most 22.5 ⁇ g, or at most 25 ⁇ g of the protein complex, wherein the protein complex comprises 60 copies (20 trimers) of DS-Cavl-I53- 50A or DS-Cavl-I53-50AAcys and 60 copies (12 pentamers) of I53-50B, I53-50B.1, 153- 50B.lNegT2 or I53-50B.4PosTl.
• the unit dose of the pharmaceutical composition comprises 1 ⁇ g, 2.5 ⁇ g, 5 ⁇ g, 7.5 ⁇ g, 10 ⁇ g, 12.5 ⁇ g, 15 ⁇ g, 17.5 ⁇ g, 20 ⁇ g, 22.5 ⁇ g, or 25 ⁇ g of the protein complex, wherein the protein complex comprises 60 copies (20 trimers) of DS- Cavl-I53-50A or DS-Cavl-I53-50AAcys and 60 copies (12 pentamers) of I53-50B, 153- 50B.1, 153-50B. lNegT2 or I53-50B.4PosTl.
• the methods of the disclosure comprise administering 1 ⁇ g, 2.5 ⁇ g, 5 ⁇ g, 7.5 ⁇ g, 10 ⁇ g, 12.5 ⁇ g, 15 ⁇ g, 17.5 ⁇ g, 20 ⁇ g, 22.5 ⁇ g, or 25 ⁇ g of the protein complex, where the protein complex comprises 60 copies (20 trimers) of DS-Cavl-I53-50A or DS-Cavl-I53-50AAcys and 60 copies (12 pentamers) of I53-50B, I53-50B.1, 153- 50B.lNegT2 or I53-50B.4PosTl.
• about 1 ⁇ g to about 5 ⁇ g, about 1 ⁇ g to about 10 ⁇ g, about 1 ⁇ g to about 15 ⁇ g, about 1 ⁇ g to about 20 ⁇ g, about 1 ⁇ g to about 25 ⁇ g, about 1 ⁇ g to about 50 ⁇ g, or about 1 ⁇ g to about 75 ⁇ g of the protein complex are administered, wherein the protein complex comprises 60 copies (20 trimers) of DS-Cavl-I53-50A or DS-Cavl-I53- 50AAcys and 60 copies (12 pentamers) of I53-50B, I53-50B.1, I53-50B.lNegT2 or 153- 50B.4PosTl.
• 1 ⁇ g to 5 ⁇ g, 1 ⁇ g to 10 ⁇ g, 1 ⁇ g to 15 ⁇ g, 1 ⁇ g to 20 ⁇ g, 1 ⁇ g to 25 ⁇ g, 1 ⁇ g to 50 ⁇ g, or 1 ⁇ g to 75 ⁇ g of the protein complex are administered, wherein the protein complex comprises 60 copies (20 trimers) of DS-Cavl-I53-50A or DS-Cavl- I53-50AAcys and 60 copies (12 pentamers) of I53-50B, I53-50B.1, 153-50B.lNegT2 or 153- 50B.4PosTl.
• about 5 ⁇ g to about 10 ⁇ g, about 5 ⁇ g to about 15 ⁇ g, about 5 ⁇ g to about 20 ⁇ g, about 5 ⁇ g to about 25 ⁇ g, about 5 ⁇ g to about 50 ⁇ g, or about 5 ⁇ g to about 75 ⁇ g of the protein complex are administered, wherein the protein complex comprises 60 copies (20 trimers) of DS-Cavl-I53-50A or DS-Cavl-I53-50AAcys and 60 copies (12 pentamers) of I53-50B, I53-50B.1, 153-50B.lNegT2 or I53-50B.4PosTl.
• 5 ⁇ g to 10 ⁇ g, 5 ⁇ g to 15 ⁇ g, 5 ⁇ g to 20 ⁇ g, 5 ⁇ g to 25 ⁇ g, 5 ⁇ g to 50 ⁇ g, or 5 ⁇ g to 75 ⁇ g of the protein complex are administered, wherein the protein complex comprises 60 copies (20 trimers) of DS-Cavl-I53-50A or DS-Cavl-I53-50AAcys and 60 copies (12 pentamers) of I53-50B, I53-50B.1, 153-50B.lNegT2 or I53-50B.4PosTl.
• about 10 ⁇ g to about 15 ⁇ g, about 10 ⁇ g to about 20 ⁇ g, about 10 ⁇ g to about 25 ⁇ g, about 10 ⁇ g to about 50 ⁇ g, or about 10 ⁇ g to about 75 ⁇ g of the protein complex are administered, wherein the protein complex comprises 60 copies (20 trimers) of DS-Cavl-I53-50A or DS-Cavl-I53-50AAcys and 60 copies (12 pentamers) of I53-50B, I53-50B.1, 153-50B. lNegT2 or I53-50B.4PosTl.
• 10 ⁇ g to 15 ⁇ g, 10 ⁇ g to 20 ⁇ g, 10 ⁇ g to 25 ⁇ g, 10 ⁇ g to 50 ⁇ g, or 10 ⁇ g to 75 ⁇ g of the protein complex are administered, wherein the protein complex comprises 60 copies (20 trimers) of DS-Cavl-I53-50A or DS-Cavl-I53-50AAcys and 60 copies (12 pentamers) of I53-50B, I53-50B.1, 153-50B.lNegT2 or I53-50B.4PosTl.
• 10 ⁇ g of the protein complex is administered, wherein the protein complex comprises 60 copies (20 trimers) of DS-Cavl-I53-50A or DS-Cavl-I53- 50AAcys and 60 copies (12 pentamers) of I53-50B, I53-50B.1, I53-50B.lNegT2 or 153- 50B.4PosTl.
• 25 ⁇ g of the protein complex is administered, wherein the protein complex comprises 60 copies (20 trimers) of DS-Cavl-I53-50A or DS-Cavl-I53- 50AAcys and 60 copies (12 pentamers) of I53-50B, I53-50B.1, I53-50B.lNegT2 or 153- 50B.4PosTl.
• 75 ⁇ g of the protein complex is administered, wherein the protein complex comprises 60 copies (20 trimers) of DS-Cavl-I53-50A or DS-Cavl-I53- 50AAcys and 60 copies (12 pentamers) of I53-50B, I53-50B.1, I53-50B.lNegT2 or 153- 50B.4PosTl.
• 100 ⁇ g of the protein complex is administered, wherein the protein complex comprises 60 copies (20 trimers) of DS-Cavl-I53-50A or DS-Cavl-I53- 50AAcys and 60 copies (12 pentamers) of I53-50B, I53-50B.1, I53-50B.lNegT2 or 153- 50B.4PosTl.
• 250 ⁇ g of the protein complex is administered, wherein the protein complex comprises 60 copies (20 trimers) of DS-Cavl-I53-50A or DS-Cavl-I53- 50AAcys and 60 copies (12 pentamers) of I53-50B, I53-50B.1, I53-50B.lNegT2 or 153- 50B.4PosTl.
• Protein complexes and pharmaceutical compositions thereof may be administered on a single dose schedule or a multiple dose schedule. Multiple doses may be used in a primary immunization schedule. In a multiple dose schedule, the various doses may be given by the same or different routes e.g., a parenteral prime and mucosal boost, a mucosal prime and parenteral boost, etc.
• the second dose of a multiple dose regimen is administered about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, or about 6 weeks after the prior dose. In embodiments, the each subsequent dose is administered 3 weeks after administration of the prior dose.
• the first dose is administered at day 0, and the second dose is administered at day 21. In embodiments, the first dose is administered at day 0, and the second dose is administered at day 28.
• Multiple doses of the boost may be used in a heterologous boost immunization schedule.
• one or more doses of a primary vaccine may be administered followed by more than one administrations of the boost vaccine.
• the various boost doses may be given by the same or different routes e.g., a parenteral prime and mucosal boost, a mucosal prime and parenteral boost, etc.
• the second dose of a multiple dose boost regimen is administered about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, or about 6 weeks after the prior dose.
• each subsequent dose is administered 3 weeks after administration of the prior dose.
• the first boost dose is administered at day 0, and the second boost dose is administered at day 21. In some embodiments, the first boost dose is administered at day 0, and the second boost dose is administered at day 28. In some embodiments, the first boost dose is administered at day 0, and the second boost dose is administered at 3 months.
• an immunogenic composition provided herein is administered as a booster of another RSV vaccine, for example, a live-attenuated RSV vaccine, an RSV- A vaccine, and RSV-B vaccine, or a bivalent RSV-A/B vaccine.
• the administering comprises administering a first dose and a second dose of the immunogenic composition, wherein the second dose is administered about 2 weeks to about 12 weeks, or about 4 weeks to about 12 weeks after the first dose is administered.
• the second dose is administered about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks, about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 9 months, about 12 months, about 18 months, about 2 years, about 3 years, about 4 years, or about 5 years after the first dose.
• three doses may be administered, with a second dose administered about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks, about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 9 months, about 12 months, about 18 months, about 2 years, about 3 years, about 4 years, or about 5 years after the first dose, and the third dose administered about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks, about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 9 months, about 12 months, about 18 months, about 2 years, about 3 years, about 4 years, or about 5 years after the second dose.
• the second dose may be an RSV booster dose.
• more than two doses of the immunogenic composition are administered.
• the first dose and the second dose of the immunogenic composition are administered within about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks, about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 9 months, or about 12 months of each other, and a third dose is administered about 1 year, about 2 years, about 3 years, about 4 years, or about 5 years after the second dose.
• the first dose and the second dose of the immunogenic composition are administered within about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks, about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 9 months, or about 12 months of each other, and subsequent doses are administered in regular intervals of about 1, 2, 3, 4 or 5 years.
• the subject has previously been infected with RSV (e.g., RSV-A and/or RSV-B).
• RSV e.g., RSV-A and/or RSV-B
• the subject is infected with RSV (e.g., RSV-A and/or RSV-B) at the time of being administered a pharmaceutical composition provided herein, wherein the administering elicits an immune response against RSV (e.g., RSV-A and/or RSV-B) in the subject that treats the RSV infection (e.g., RSV-A infection and/or RSV-B infection) in the subject.
• RSV infection e.g., RSV-A infection and/or RSV-B infection
• the immunogenic compositions are administered to a subject that has already been infected with RSV (e.g., RSV-A and/or RSV-B), and/or who is suffering from symptoms (such as described above) indicating that the subject is likely to have been infected with RSV (e.g., RSV-A and/or RSV-B).
• RSV infection e.g., RSV-A infection and/or RSV-B infection
• RSV infection may be diagnosed using any PCR-based test or antigen-based test known in the art.
• the subject has antibodies against RSV.
• Anti-RSV antibodies may be detected using any serological test known in the art.
• the compositions and methods disclosed herein prevent disease following infection with RSV subtypes A and B in older adults.
• a method comprises administering a protein complex or pharmaceutical composition thereof about 2 weeks to about 12 weeks, or about 4 weeks to about 12 weeks after another vaccine, such as a heterologous prime vaccine.
• the pharmaceutical composition is administered about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks, about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 9 months, about 12 months, about 18 months, about 2 years, about 3 years, about 4 years, or about 5 years after the other vaccine.
• the protein complex or pharmaceutical composition thereof is administered about 2 or more months, about 3 or more months, about 4 or more months, about 5 or more months, about 6 or more months, about 8 or more months, about 10 or more months, or about 12 or months after an earlier vaccine.
• a method comprises administering a protein complex or pharmaceutical composition thereof about 2 months to about 8 months, or about 2 months to about 6 months after another vaccine.
• the interval between first (prime) vaccine and second (boost) vaccine may be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or any other suitable interval.
• the prime vaccine may include multiple doses of the same vaccine, and the heterologous boost vaccine may include multiple doses of the same heterologous vaccine, administered at suitable intervals.
• the method may comprise administering a protein complex or pharmaceutical composition thereof indefinitely, e.g., over regular intervals.
• the regular intervals may include every 3 months, every 6 months, every 12 months, every 18 months, or every 24 months.
• the polypeptide sequence of the antigen may be modified to compensate for antigenic drift.
• the protein complexes and pharmaceutical compositions of the disclosure may also be used for homologous prime-boost vaccination (e.g., administering a booster dose following a primary regimen of the same vaccine).
• a method comprises administering a protein complex or pharmaceutical composition thereof about 2 weeks to about 12 weeks, or about 4 weeks to about 12 weeks after another vaccine, such as a heterologous prime vaccine.
• the pharmaceutical composition is administered about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks, about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 9 months, about 12 months, about 18 months, about 2 years, about 3 years, about 4 years, or about 5 years after the other vaccine.
• the protein complex or pharmaceutical composition thereof is administered about 2 or more months, about 3 or more months, about 4 or more months, about 5 or more months, about 6 or more months, about 8 or more months, about 10 or more months, or about 12 or months after an earlier vaccine.
• a method comprises administering a protein complex or pharmaceutical composition thereof about 2 months to about 8 months, or about 2 months to about 6 months after another vaccine.
• the interval between first (prime) vaccine and second (boost) vaccine may be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or any other suitable interval.
• the prime vaccine may include multiple doses of the same vaccine, and the homologous boost vaccine may include multiple doses of the homologous vaccine, administered at suitable intervals.
• the method comprises administering a protein complex or pharmaceutical composition thereof continuously, e.g., over regular intervals.
• the regular intervals may include every 3 months, every 6 months, every 12 months, every 18 months, or every 24 months.
• the disclosure further provides prime-boost strategies that employ any known or subsequently developed vaccine - including but not limited to a protein, DNA, mRNA, inactivated virus, or viral vector vaccine - together with a protein complex or pharmaceutical composition as described herein.
• the protein complexes described herein may be used as a primary vaccine followed by heterologous boost with another vaccine.
• the subject may receive a further vaccination with a protein complex described herein.
• another vaccine is used as the primary vaccine and a protein complex described herein is administered one or more times to boost the response to the primary vaccine.
• kits which may be used to prepare the virus-like particles and compositions of the disclosure.
• a kit provided herein comprises a first component and a second component as disclosed herein, and instructions for use in a method of the disclosure.
• a kit comprises one or more unit doses as disclosed herein, and instructions for use in a method of the disclosure.
• the kit comprises a vial comprising a single dose of a pharmaceutical composition provided herein.
• a kit comprises a vial comprising multiple doses provided herein.
• a kit further comprises instructions for use of the pharmaceutical composition.
• kits further comprises a diluent for preparing dilutions of the pharmaceutical composition prior to administration.
• the pharmaceutical composition comprises an adjuvant.
• a kit comprises a pharmaceutical composition and an adjuvant which must be mixed prior to administration.
• a pharmaceutical composition comprising a protein complex comprising a first component comprising an RSV F protein and a first multimerization domain; and one or more pharmaceutically acceptable diluents or excipients.
• compositions of any one of embodiments 1 to 14, wherein the RSV F protein comprises an amino acid sequence which is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence of any one of SEQ ID NOs: 14, 34, and 35.
• the second component comprises an amino acid sequence which is at least 75%, 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 26.
• the unit dose of embodiment 20, wherein the unit dose comprises between about 25 ⁇ g and about 250 ⁇ g of the protein complex; between about 5 pmol and about 50 pmol of the protein complex; and/or between about 100 pmol and about 1,000 pmol of the RSV F protein.
• the unit dose of embodiment 20, wherein the unit dose comprises between about 25 ⁇ g and about 75 ⁇ g of the protein complex; between about 5 pmol and about 15 pmol of the protein complex; and/or between about 100 pmol and about 250 pmol of the RSV F protein.
• the unit dose of embodiment 20, wherein the unit dose comprises between about 75 ⁇ g and about 250 ⁇ g of the protein complex; between about 15 pmol and about 50 pmol of the protein complex; and/or between about 250 pmol and about 1,000 pmol of the RSV F protein.
• the unit dose of embodiment 20, wherein the unit dose comprises about 25 ⁇ g, about 75 ⁇ g, or about 250 ⁇ g of the protein complex; about 5 pmol, about 15 pmol, or about 50 pmol of the protein complex; and/or about 100 pmol, about 250 pmol, or about 1,000 pmol of the RSV F protein.
• a method of treating and/or preventing severe lower respiratory tract infection (LRTI) associated caused by RSV in a subject comprising administering to the subject an effective amount of the pharmaceutical composition of any one of embodiments 1 to 19.
• LRTI severe lower respiratory tract infection
• a method of preventing RSV disease in a subject comprising administering to the subject an effective amount of the pharmaceutical composition of any one of embodiments 1 to 19.
• [0190] 34 The method of embodiment 33, wherein the subject is at risk of severe RSV disease because of underlying diabetes mellitus, cardiovascular disease, or respiratory disease.
• a method of generating an immune response in an unborn child comprising administering an effective amount of the pharmaceutical composition of any one of embodiments 1 to 19 to the mother of said unborn child.
• [0200] 44 A method of generating an immune response in an infant and/or prevent respiratory syncytial virus (RSV) disease in an infant through maternal immunization of a pregnant subject , the method comprising administering an effective amount of the pharmaceutical composition of any one of embodiments 1 to 19 to the subject.
• RSV respiratory syncytial virus
• [0205] 49 The method of embodiment 45, wherein the effective amount is about 25 ⁇ g, about 75 ⁇ g, or about 250 ⁇ g of the protein complex; about 5 pmol, about 15 pmol, or about 50 pmol of the protein complex; and/or about 100 pmol, about 250 pmol, or about 1,000 pmol of the RSV F protein.
• [0206] 50 The method of embodiment 45, wherein the effective amount is at least about 25 ⁇ g, at least about 75 ⁇ g, or at least about 250 ⁇ g of the protein complex; at least about 5 pmol, at least about 15 pmol, or at least about 50 pmol of the protein complex; and/or at least about 100 pmol, at least about 250 pmol, or at least about 1,000 pmol of the RSV F protein.
• [0207] 51 The method of embodiment 45, wherein the effective amount is at most about 25 ⁇ g, at most about 75 ⁇ g, or at most about 250 ⁇ g of the protein complex; at most about 5 pmol, at most about 15 pmol, or at most about 50 pmol of the protein complex; and/or at most about 100 pmol, at most about 250 pmol, or at most about 1,000 pmol of the RSV F protein.
• [0210] 54 The method of embodiment 53, wherein the second dose is administered within about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks, about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 9 months, about 12 months, about 24 months, or about 36 months of the first dose.
• [0212] 56 The method of embodiments 55, wherein the third dose is administered about 1 year, about 2 years, about 3 years, about 4 years, about 5 years, about 6 years, about 7 years, about 8 years, about 9 years, or about 10 years after the second dose.
• [0220] 64 The method of embodiment 63, wherein the method results in an increase in RSV-B-specific neutralizing antibodies in the subject of at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 8-fold, at least about 10- fold, at least about 15-fold, at least about 20-fold, or at least about 25-fold compared to baseline.
• any one of embodiments 28 to 84 wherein the method results in protective immunity for a longer time period than a method involving administration of a trimeric antigen having an amino acid sequence which is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence of any one of SEQ ID NOs: 7, 14, 34, and 35.
• Example 1 Characterization of Immune Responses Induced by an RSV Vaccine in Naive Mice
• the candidate RSV vaccine was administered intramuscularly at three dose levels (8.33 ⁇ g, 2.5 ⁇ g and 0.83 ⁇ g) either unadjuvanted, adjuvanted with Alhydrogel (aluminum hydroxide adjuvant), or adjuvanted with Addavax (oil-in-water emulsion) on Day 0, Day 21 and Day 42. Serum samples were obtained on Days 0, 42 and 56 to measure neutralization antibody titers using a virus neutralization assay. Clinical observations were made daily and animals were weighed weekly.
• mice Two hundred female Balb/c mice, 6-8 weeks of age, were infected with IxlO 6 pfu RSV A2 intranasally and housed for twelve weeks to allow for the resolution of the infection and the establishment of immunological memory. Twenty mice were not infected and served as naive controls. [0278] Animals were allocated to 19 experimental groups according to neutralizing antibody titers from Day 28 serum samples to obtain comparable groups. The mice were vaccinated intramuscularly on Day 91 with one of four different dose levels of the candidate RSV vaccine (1.66, 0.5, 0.16 and 0.016 ⁇ g) or an equivalent amount of a stabilized RSV F- protein (1, 0.3, 0.1, and 0.01 ⁇ g) (Table 3).
• RSV neutralization titers were measured on Days 0, 28, 87, and 101. Serum samples were collected from each animal prior to infection on Day 0, Day 28, Day 87 and Day 101 (Table 4). Splenocytes were isolated from a subset of animals on Days 101 and 102 and stimulated with RSV-specific peptides in an ELISpot assay or a cytokine release assay (a cell-mediated immunity assay).
• This study was designed to explore the safety and immunogenicity of up to 3 vaccinations of soluble RSV vaccine (in Addavax, oil-in-water emulsion) and aluminum hydroxide (Alhydrogel)-adjuvanted formulations in rabbits.
• the purpose of this study was to develop an initial assessment of the safety of and RSV vaccine in rabbits; to assess the immune response in vaccinated animals; and to assess whether the presence of pre-existing antibodies to the VLP core (i.e., the protein complex lacking the RSV F-protein) interferes with the response to the RSV vaccine in rabbits.
• NZW New Zealand White
• mice Five New Zealand White (NZW) female rabbits (Group 1) were administered 0.5 mg of VLP core with Addavax by IM injection on Days 1 and 14. Subsequently, the rabbits in Group 1 were vaccinated with 0.25 mg RSV vaccine adsorbed to 0.5 mg aluminum hydroxide adjuvant on Days 56, 70 and 84. Two additional groups of 3 female rabbits each were vaccinated on Days 56, 70 and 84 with 0.25 mg RSV vaccine alone (Group 2) or 0.25 mg adsorbed to 0.5 mg aluminum hydroxide adjuvant (Group 3) with no VLP core prior administration. All vaccinations consisted of a total volume of 0.5 mL split into two 0.25 mL injections (injection on the right and left thigh).
• NZW rabbits were vaccinated (0.5 mL/dose) with sterile saline (negative control), 0.25 mg RSV vaccine alone or 0.25 mg RSV vaccine adsorbed to 0.5 mg aluminum hydroxide adjuvant once every 2 weeks for atotal of three vaccinations (i.e., administrations on Day 1, 15 and 29).
• the volume per injection and the dose level of RSV vaccine without adjuvant and with aluminum hydroxide adjuvant are equivalent to the intended clinical dose and injection volume.
• Three days following the third vaccination 5 animals/sex were sacrificed (on Day 32); the remaining animals (5/sex/group) were maintained for an additional 4 weeks and sacrificed on recovery day (RD) 29 (Study Day 61). See Table 5 for group allocation.
• RSV vaccine contained 0.5 mg/mL RSV vaccine in 20 mM Tris, 200 mM NaCl, 4% sucrose, pH 7.8 ⁇ 0.2 with or without 1 mg/mL aluminum hydroxide adjuvant.
• Example 5 A Phase 1/lb (Phl/lb) Study to Evaluate the Safety and Immunogenicity of an RSV Vaccine in Healthy Adults
• This example describes a Phasel/lb Study is a randomized placebo-controlled observer-blind study to assess the safety and immunogenicity of a single intramuscular (IM) dose of RSV vaccine as aqueous or aluminum hydroxide-adjuv anted formulations.
• the trial design is shows in FIG. 4.
• a total of six formulations (three dose levels of the aqueous formulation and three dose levels of the aluminum hydroxide-adjuvanted formulation) were tested. Placebo was administered as a control.
• the study was conducted in two parts:
• the duration of subject participation in the study will be approximately 6 months.
• the three dose levels tested in the study 25 ⁇ g, 75 ⁇ g, and 250 ⁇ g of the VLP protein complex, are equivalent in mass to 14 ⁇ g, 42 ⁇ g, and 140 ⁇ g, respectively, of a soluble RSV F antigen — based on subtracting the mass of the VLP core from the mass of the complete VLP including both displayed antigens and its core.
• SAEs Serious adverse events
• MAAEs medically attended adverse events
• AEs adverse events leading to study withdrawal
• SRR seroresponse rate
• GMFR geometric mean fold rise
• Investigational vaccine was formulated at one concentration, 250 ⁇ g/0.5 mL as an aqueous vaccine or adsorbed to 500 ⁇ g aluminum hydroxide as an adjuvant.
• the lower dosage vaccines (25 and 75 ⁇ g) for each formulation were dilutions of the highest dose and were prepared just prior to administration using either aqueous or aluminum hydroxide adjuvant diluent.
• the aluminum hydroxide content was the same for all adjuvant formulations. All RSV vaccine formulations were administered as 0.5 mL doses.
• Co-Primary Endpoints Based on RSV-A-specific NT Abs at Day 28: SRR (percentage of subjects with >4-fold rise in titer versus baseline (Day 0)) and GMFR versus baseline (Day 0).
• GMFR in titers versus baseline (Day 0; GMFR) at Days 7 and 180; SRRs to either RSV strain at Days 7 and 180; Percentages of subjects with >4-fold rise in titers versus baseline to either RSV strain at Days 7, 28 and 180; Percentages of subjects with >8-fold rise in titers versus baseline to either RSV strain at Days 7, 28 and 180; SRRs to both RSV strains at Days 7, 28 and 180; Geometric mean titer (GMT) to both RSV strains at Days 0, 7, 28 and 180.
• This example describes a study to assess the immunogenicity out to 12 months from the single dose given in the study in Example 5 as well as the effect of revaccination with RSV vaccine.
• RSV vaccine is administered to up to 120 older adults (e.g., adults of over 60 years of age) at a 75 ⁇ g unadjuvanted IVX-121 dose. These subjects may be a subset of the older adult study population described in Example 5 infra. Subjects are revaccinated 12 months after the initial dose and efficacy and safety are assessed six months after the revaccination dose. Safety and efficacy endpoints are those described in Example 5.
• Example 5 provides interim results from Example 5: a Phase 1/lb clinical trial of IVX-121, a VLP displaying a prefusion stabilized Respiratory Syncytial Virus (RSV) F antigen, in young and older adults.
• IVX-121 demonstrated a robust immunologic response in both young and older adult groups.
• RSV Respiratory Syncytial Virus
• the Phase 1/lb clinical trial of IVX-121 is a randomized, observer-blinded, placebo- controlled, multi-center study designed to evaluate the safety and immunogenicity of three dose levels of IVX-121, with and without aluminum hydroxide adjuvant, in healthy young and older adults.
• the study design is shown in FIG. 4.
• Subjects were administered a single dose of IVX-121 at one of three dose levels (25, 75, 250 ⁇ g), with or without aluminum hydroxide adjuvant, or placebo.
• the primary outcomes of the study were safety and immunogenicity up to 28 days post-vaccination; neutralizing antibodies to RSV-A and RSV-B were measured in international units (lU/mL) using the WHO international reference standard..
• IVX-121 induced a robust immune response in both young and older adult groups. Data indicated a dose-independent response, including at the lowest non-adjuvanted dose (25 ⁇ g) (FIG. 7). No additional benefit from the aluminum hydroxide adjuvant was observed at any dosage level in either portion of the study (FIG. 8). Geometric mean titers for RSV-A and RSV-B were in comparable ranges for both groups (FIG. 9)
• IVX-121 was generally well tolerated and elicited a strong and consistent response against RSV in healthy young and older adults. These data are particularly encouraging for the vulnerable older adult population with co-morbidities and increased risk for severe disease and hospitalization. IVX-121 exhibits immunogenicity at very low microgram dosage levels and tolerability to the highest dose level. Eligible older adults from the Phase lb cohort will be followed out to 12 months to assess durability of response.
• RSV/A and RSV/B neutralizing antibody (NAb) assays were used to measure the presence of RSV-specific neutralizing antibodies. These assays were been validated for standard bioanalytical assay parameters to include Specificity, Accuracy, Precision, Repeatability (intra-assay precision), Intermediate Precision, Linearity, Dilutional Linearity, Range, Stability, and Robustness. Critical reagents for these assays are commercially purchased and qualified for use. In brief, human serum samples are serially diluted and incubated with a constant concentration of virus (RSV A2 (ATCC, Cat# VR- 1540) and RSV B 18537 (ATCC, Manassas, VA; Cat# VR-1580)).
• RSV A2 ATCC, Cat# VR- 1540
• RSV B 18537 ATCC, Manassas, VA; Cat# VR-1580
• Neutralization is measured by the inhibition of virus propagation in HEp-2 cells (ATCC, Cat# CCL23). Following an incubation period, cells are fixed and immunostained with a murine monoclonal antibody directed against RSV F protein, followed by horse radish peroxidase (HRP)-conjugated goat-anti-mouse antibody and TrueBlue (TB). The plates are scanned with an UV Analyzer and Spot counts (SC; or spot forming cells, SCF) per well at each serum/antibody concentration are quantified. The values are then analyzed to determine the dilution of serum/antibody that yield a selected reduction point (z.e., 50% or IC50).
• HRP horse radish peroxidase
• SCF spot forming cells
• the RSV Reference Standard (16/284) with an assigned a potency of 1000 International Units (IU) anti-RSV/A or anti-RSV/B neutralizing antibodies per ampoule was used to establish and validate an IHR to be included during neutralization titer analyses.
• the potency of the IHR standard was determined to be 1,831 lU/mL for RSV/A and 609 lU/mL for RSV/B. These potencies were used to calculate conversion factors which can be used for converting MN titer results (AU/mL) to lU/mL.
• the potency of a test sample in lU/mL was calculated by multiplying the test sample MN titer by 1.0578 for RSV/A and by 0.6936 for RSV/B.
• the RSV/A and RSV/B NAb assays has been validated to an assay range of 9.4 - 180263 AU/mL and 8 - 195392 AU/mL, respectively.
• the validated assay range in lU/mL for RSV/A and RSV/B are 9.9 - 190682 and 5.5 - 135524, respectively.
• IVX-121 is formulated as a non-adjuvanted vaccine.
• the vaccine is administered in a single dose of at most about 25 ⁇ g, such as 1 ⁇ g, 2.5 ⁇ g, 5 ⁇ g, 7.5 ⁇ g, 10 ⁇ g, 12.5 ⁇ g, 15 ⁇ g, 17.5 ⁇ g, 20 ⁇ g, 22.5 ⁇ g, or 25 ⁇ g of the IVX-121 protein complex.
• Subjects may be selected as adults 18+ years of age or adults 60+ years of age, or as individuals at high risk of severe disease (e.g, affected by underlying chronic conditions, including diabetes mellitus, cardiovascular diseases, and respiratory diseases; frail elderly; or immunocompromised).
• Clinical endpoints include RSV-associated LRTD (lower respiratory tract disease), moderate/severe RSV-associated LRTD, and/or RSV-associated acute respiratory disease.
• Subjects are monitored for one, two, or three RSV seasons. The vaccine may be administered again after three to five years.

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