WO2022260960A1 - Virus-like particle vaccine for coronavirus - Google Patents

Virus-like particle vaccine for coronavirus Download PDF

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Publication number
WO2022260960A1
WO2022260960A1 PCT/US2022/032201 US2022032201W WO2022260960A1 WO 2022260960 A1 WO2022260960 A1 WO 2022260960A1 US 2022032201 W US2022032201 W US 2022032201W WO 2022260960 A1 WO2022260960 A1 WO 2022260960A1
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Prior art keywords
sars
protein
subject
cov
vaccine
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PCT/US2022/032201
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English (en)
French (fr)
Inventor
Niranjan Kanesa-Thasan
Charles Richardson
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Icosavax Inc
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Icosavax Inc
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Priority to MX2023014468A priority Critical patent/MX2023014468A/es
Priority to CN202280055149.2A priority patent/CN118159288A/zh
Priority to AU2022289462A priority patent/AU2022289462A1/en
Priority to US18/565,728 priority patent/US20240252621A1/en
Priority to JP2023574797A priority patent/JP2024520730A/ja
Priority to KR1020247000038A priority patent/KR20240019213A/ko
Priority to EP22820815.3A priority patent/EP4351638A4/en
Priority to CA3221041A priority patent/CA3221041A1/en
Priority to IL308842A priority patent/IL308842A/en
Publication of WO2022260960A1 publication Critical patent/WO2022260960A1/en
Anticipated expiration legal-status Critical
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/12—Viral antigens
    • A61K39/215—Coronaviridae, e.g. avian infectious bronchitis virus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/02—Bacterial antigens
    • A61K39/025—Enterobacteriales, e.g. Enterobacter
    • A61K39/0258—Escherichia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/12—Viral antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/525—Virus
    • A61K2039/5258—Virus-like particles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/545—Medicinal preparations containing antigens or antibodies characterised by the dose, timing or administration schedule
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55505—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
    • A61K2039/55561—CpG containing adjuvants; Oligonucleotide containing 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
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • 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
    • A61K2039/60—Medicinal preparations containing antigens or antibodies characteristics by the carrier linked to the antigen
    • A61K2039/6031—Proteins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/60—Medicinal preparations containing antigens or antibodies characteristics by the carrier linked to the antigen
    • A61K2039/6031—Proteins
    • A61K2039/6068—Other bacterial proteins, e.g. OMP
    • 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
    • C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
    • C12N2770/00011—Details
    • C12N2770/20011—Coronaviridae
    • C12N2770/20034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein

Definitions

  • the present disclosure relates to targeting SARS-CoV-2, in particular, prevalent strains of SARS-CoV-2, and methods of using such vaccines to induce neutralizing antibody levels against SARS-CoV-2.
  • Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a viral pathogen responsible for the coronavirus disease 2019 (COVID-19) global pandemic. As of May 2022, there were over 500 million cumulative cases and over 6.2 million deaths from COVID-19 worldwide with over 1 million deaths in the United States alone. Rates of serious morbidity and mortality from COVID-19 are disproportionately higher in older adults as compared to other age groups, likely due to age-induced immunosenescence. Despite the fact that adults over 65 constitute 17% of the United States population, over 75% of the deaths in the United States due to COVID-19 have been in this age group.
  • Vaccines have been developed to combat this pandemic at an unprecedented pace and there are several SARS-CoV-2 vaccines that have been licensed or approved under emergency use authorization.
  • the initial push for first wave vaccines to fight the pandemic has focused on speed rather than other critical attributes that are now important considerations for second wave vaccine candidates such as durability, potential to boost response, potential to address variant strains, ease of manufacturing and distribution, stability, and reactogenicity profile.
  • Coronaviruses are prone to mutation but the pace at which the SARS-CoV-2 virus has mutated is faster than most were anticipating. Some of these emerging strains appear to enhance transmission and pathogenicity, with complete replacement of the original SARS-CoV-2 strain by the emerging strains in some countries. Data has shown that some vaccines against the original SARS-CoV-2 virus strain are less immunogenic against some of the emerging variants, particularly the B.1.351 (beta) and B.1.1.529 (omicron) variants first identified in South Africa. Decreases in neutralizing titers against the B.1.351 and B.1.1.529 strains in vitro appear to translate to lower efficacy in people who are infected with these virus strains.
  • VLP vaccines allow for stable multivalent antigen display, facilitating cross-linking of B-cell receptors and driving stronger immune signaling than soluble protein antigens.
  • VLP vaccines have historically been shown to induce durable immunity [ e.g . human papilloma virus (HPV)] and there are several examples of licensed vaccines utilizing naturally- occurring self-assembling VLPs, including human papilloma virus (HPV) and hepatitis B (HBV) vaccines.
  • HPV human papilloma virus
  • HBV hepatitis B
  • compositions and methods of the present disclosure address that need.
  • a pharmaceutical composition comprising a protein complex comprising a first component comprising a receptor-binding domain of a coronavirus S protein and a first multimerization domain, and optionally a second component comprising a second multimerization domain; and one or more pharmaceutically acceptable diluents or excipients.
  • the pharmaceutical composition comprises an adjuvant.
  • the adjuvant is a squalene-in-water emulsion.
  • the adjuvant is MF59 ® .
  • the adjuvant comprises an oil-in-water emulsion.
  • the protein complex is an icosahedral protein complex.
  • 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 selected from the group consisting of SEQ ID NOS: 9-13 or 18.
  • 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 the group consisting of SEQ ID NOS: 14-17, 20 or 27.
  • 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 any one of SEQ ID NOs: 1-6; 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: 14.
  • a unit dose of the pharmaceutical composition described herein wherein the unit dose comprises 2 pg, 5 pg, 10 pg, 15 pg, 25 pg, 50 pg, 100 pg, or 125 pg 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 pg and about 125 pg of the protein complex.
  • the unit dose of the pharmaceutical composition is between about 2 pg to about 125 pg, or between about 5 pg to about 125 g, or between about 15 pg to 125 pg, or between about 25 pg to about 125 pg, or between about 50 pg to about 125 pg, or between about 100 pg to about 125 pg of the protein complex.
  • the disclosure provides a pharmaceutical composition, comprising a protein complex comprising a first component comprising a receptor-binding domain of a coronavirus S protein attached to a first multimerization domain, and optionally a second component comprising a second multimerization domain; and optionally one or more pharmaceutically acceptable diluents or excipients.
  • the pharmaceutical composition comprises an adjuvant.
  • the adjuvant is a squalene-in-water emulsion.
  • the adjuvant is MF59®.
  • the adjuvant is an aluminum salt.
  • the adjuvant is CPG-1018.
  • the pharmaceutical composition comprises both an aluminum salt and CPG-1018.
  • the pharmaceutical composition is free of or substantially free of any adjuvant.
  • the first multimerization domain is a trimerization domain and the second multimerization domain is a pentamerization domain.
  • the protein complex is an icosahedral protein complex.
  • 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 selected from the group consisting of SEQ ID NOS: 9- 13 or 18.
  • 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 the group consisting of SEQ ID NOS: 14- 17, 20 or 27.
  • 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 any one of SEQ ID NOs: 1-6; and wherein 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: 14.
  • the disclosure provides a unit dose of the pharmaceutical composition of any one of embodiments 1 to 13, wherein the unit dose comprises 2 pg, 5 pg, 10 pg, 15 pg, 25 pg, 50 pg, 100 pg, or 125 pg of the protein complex.
  • the disclosure provides a method of vaccinating a subject at risk of infection with SARS-CoV-2, comprising administering to the subject a pharmaceutical composition comprising an effective amount of a protein complex comprising a first component comprising a receptor-binding domain of a coronavirus S protein attached to a first multimerization domain, and a second component comprising a second multimerization domain; and one or more pharmaceutically acceptable diluents or excipients.
  • the disclosure provides a method of boosting an immune response to a prior vaccination for SARS-CoV-2, comprising administering to a subject previously vaccinated for SARS-CoV-2 a pharmaceutical composition comprising an effective amount of a protein complex comprising a first component comprising a receptor-binding domain of a coronavirus S protein attached to a first multimerization domain, and optionally a second component comprising a second multimerization domain.
  • the subject has been previously vaccinated with a full vaccination course of a primary vaccine.
  • the disclosure provides a method of safely and effectively immunizing a subject for SARS-CoV-2, comprising administering to a subject previously vaccinated for SARS-CoV-2 a pharmaceutical composition comprising an effective amount of a protein complex comprising a first component comprising a receptor-binding domain of a coronavirus S protein attached to a first multimerization domain, and optionally a second component comprising a second multimerization domain.
  • the pharmaceutical composition comprises an adjuvant.
  • the adjuvant is a squalene-in-water emulsion.
  • the adjuvant is MF59®.
  • the adjuvant is an aluminum salt.
  • the adjuvant is CPG-1018.
  • the pharmaceutical composition comprises both an aluminum salt and CPG-1018.
  • the pharmaceutical composition is free of or substantially free of any adjuvant.
  • the first multimerization domain is a trimerization domain and the second multimerization domain is a pentamerization domain.
  • the protein complex is an icosahedral protein complex.
  • 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 selected from the group consisting of SEQ ID NOS: 9- 13 or 18.
  • 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 the group consisting of SEQ ID NOS: 14- 17, 20 or 27.
  • 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 any one of SEQ ID NOs: 1-6; and wherein 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: 14.
  • the effective amount is 2 mg, 5 pg, 10 pg, 15 pg, 25 pg, 50 pg, 100 pg, or 125 pg of the protein complex.
  • the method comprises repeating the administering step.
  • the method comprises administering a booster vaccine.
  • the method comprises administering a prime vaccine.
  • the prime vaccine is an mRNA-based vaccine, an adenoviral vector- based vaccine, a protein— based vaccine, or an inactivated virus vaccine.
  • the prime vaccine is the protein complex.
  • the subject is a previously vaccinated subject.
  • the subject has completed a full course of vaccination for an original strain of SARS-CoV-2.
  • the subject has completed a partial course (e.g., has received one of two doses) of vaccination for an original strain of SARS-CoV-2.
  • the subject has received at least one dose of a vaccination for a variant strain of SARS-CoV-2.
  • the subject has received at least one dose of a vaccine comprising the receptor binding domain of a coronavirus S protein or a polynucleotide encoding the receptor binding domain of a coronavirus S protein.
  • the subject has received at least one dose of a vaccine comprising a coronavirus S protein or a polynucleotide encoding a coronavirus S protein.
  • the coronavirus S protein is S2P.
  • the S protein is HexaPro.
  • the subject is a vaccination naive subject.
  • the subject has previously been infected with SARS-CoV-2.
  • the subject has not previously been infected with SARS-CoV-2. [0059] In some embodiments, the subject does not have antibodies against SARS-CoV-2 prior to the administering step.
  • the subject has antibodies against SARS-CoV-2 prior to the administering step.
  • the method induces neutralizing antibody titers in the subject.
  • the method induces S protein-specific and IgG antibody titers in the subject.
  • the method prevents infection with SARS-CoV-2.
  • the method prevents infection with an original strain of SARS-CoV-
  • the method prevents infection with a variant strain of SARS-CoV-
  • the method reduces the severity of infection with coronavirus.
  • the method reduces the severity of infection with an original strain of SARS-CoV-2.
  • the method reduces the severity of infection with a variant strain of SARS-CoV-2.
  • FIG. 1 shows a structural model of assembly of a vaccine from a first component including an antigenic fragment (here: the receptor binding domain) of the S protein (CompA-RBD-01) and a second component (CompB).
  • a first component including an antigenic fragment (here: the receptor binding domain) of the S protein (CompA-RBD-01) and a second component (CompB).
  • FIG. 2 shows a summary of a clinical trial design.
  • FIG. 3 is a schematic showing an IVX-411 Phase 1/2 trial study overview.
  • the topline data includes two components.
  • FIG. 4 is a schematic showing an IVX-411 Phase 1/2 study design.
  • FIGs. 5A and 5B are graphs showing local (FIG. 5A) and systemic (FIG. 5B) adverse events (AEs) within 7 days of any dose in Parts 1 and 2 of the study.
  • FIGs. 6A and 6B are graphs showing neutralizing and spike IgG antibody titers in Part 1 - SARS-CoV-2-nai ' ve subjects (FIG. 6A) and Part 2 - previously vaccinated subjects (FIG. 6B) of the study.
  • FIGs. 7A and 7B are graphs showing wild type and omicron neutralizing antibody titers in Part 1 - SARS-CoV-2 naive subjects (FIG. 7A) and Part 2 - previously vaccinated subjects (FIG. 7B) of the study.
  • compositions comprising a protein complex comprising which may be used in the treatment of SARS-CoV2.
  • 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 as calculated by 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).
  • 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 ., COVID19) using a vaccine made with another technology (e.g., an mRNA vaccine, adenoviral vector vaccine, or a protein based 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., COVID19) using a vaccine made with another technology (e.g., an mRNA vaccine, adenoviral vector vaccine, or a protein based 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 based 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., COVID19) 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., COVID19), 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., COVID19) 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.
  • the term “HexaPro” refers to a S protein four beneficial proline substitutions (F817P, A892P, A899P, A942P) as well as the two proline substitutions in S-2P (prolines at positions 986 and 987). See Hsieh et al. Science 369:1501-05 (2020).
  • the subject is a vaccination naive or SARS-CoV-2 uninfected subject.
  • the vaccine is an mRNA-based vaccine, an adenoviral vector-based vaccine, a protein-subunit based vaccine, or an inactivated virus vaccine.
  • a “subunit” composition for example a vaccine, that includes one or more selected antigens but not all antigens from a pathogen.
  • 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.
  • Illustrative designed VLP are VLPs that comprise nanostructures depicted in FIG. 1.
  • the term “symmetric VLP” refers to a protein-based VLP with a symmetric core, such as shown in FIG. 1. 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 VLPs 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.
  • template structures e.g., structures deposited in the Protein Data Bank
  • 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.
  • the subject is a human.
  • 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).
  • the subject is up to 4 months old, or up to 6 months old.
  • the adults are seniors about 65 years or older, or about 60 years or older.
  • 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 (e.g., a protein-based Virus-like Particle) comprising a first component comprising a receptor binding domain of a coronavirus spike (S) protein, or alternatively another antigenic portion of the coronavirus S protein, and a first multimerization domain.
  • a protein complex e.g., a protein-based Virus-like Particle
  • SARS-CoV-2 severe acute respiratory syndrome coronavirus 2
  • the genetic sequence of SARS-CoV-2 became available to the WHO and public (MN908947.3) and the virus was categorized into the betacoronavirus subfamily.
  • SARS severe acute respiratory syndrome
  • MERS Middle East respiratory syndrome
  • Coronaviruses are positive-sense, single-stranded RNA ((+)ssRNA) enveloped viruses that encode for a total of four structural proteins, spike protein (S), envelope protein (E), membrane protein (M) and nucleocapsid protein (INI).
  • S protein spike protein
  • E envelope protein
  • M membrane protein
  • II nucleocapsid protein
  • S protein spike protein
  • S protein is responsible for receptor-recognition, attachment to the cell, infection via the endosomal pathway, and the genomic release driven by fusion of viral and endosomal membranes. Though sequences between the different family members vary, there are conserved regions and motifs within the S protein making it possible to divide the S protein into two subdomains: SI and S2.
  • S1 domain recognizes the virus- specific receptor and binds to the target host cell.
  • the structure of the SARS-CoV-2 S protein, including its receptor binding domain (RBD), has been determined by cyro-electron microscopy (Cyro-EM) (Wrapp et al. Science 367:1260-1263 (2020)).
  • the S 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 antigenic portion of the first component may comprise, consist essentially of, or consist of a selected fragment of the coronavirus S protein.
  • the antigenic portion may comprise the receptor binding domain of the coronavirus S protein with flanking sequences on the domain’s N or C terminus ( e.g ., 5, 10, 20, 30, or more amino acids of the coronavirus S protein outside the receptor binding domain); or the antigenic portion may include only a few flanking amino acids (e.g., 1, 2, 3, 4, or 5 amino acids from the coronavirus S protein); or the antigenic portion may include only the receptor binding domain with no flanking sequences from the coronavirus S protein.
  • flanking sequences on the domain’s N or C terminus e.g ., 5, 10, 20, 30, or more amino acids of the coronavirus S protein outside the receptor binding domain
  • flanking amino acids e.g., 1, 2, 3, 4, or 5 amino acids from the coronavirus S protein
  • the antigenic portion may include only the receptor binding domain with no flanking sequences from the coronavirus S protein.
  • 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.
  • the “core” of the VLP is used herein to describe the central portion of the VLP that links together the several copies of the RBD or coronavirus S protein ectodomain, or antigenic fragments thereof, displayed by the VLP.
  • the first component comprises a first polypeptide comprising an RBD, a linker, and a first polypeptide comprising a multimerization domain.
  • the VLP is adapted to display the RBD or S protein from two or more diverse strains of coronavirus.
  • the same VLP displays mixed populations of protein antigens or mixed heterotrimers of protein antigens from different strains of coronavirus.
  • the 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 anN- 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.
  • an VLP is created using a cleavable or non-cleavable linker.
  • Processes and methods for conjugation of antigens to carriers are provided by, e.g., U.S. Patent Pub. No. US 2008/0145373 Al.
  • 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 A1 describes various methods for designing protein assemblies. As described in US Patent Pub No. US 2016/0122392 A1 and in International Patent Pub. No.
  • 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.
  • the protein complex is a designed protein-based VLP as depicted in FIG. 1.
  • the protein-based VLP may comprise the proteins described in Table 3 or functional variants thereof.
  • the VLP may display the receptor-binding domain of a coronavirus spike (S) protein, such as SARS-CoV-2, or it may display an ectodomain of a coronavirus S protein. While certain representative protein-based VLPs are described herein, in variations, other protein-based VLPs may be used.
  • the VLP may be a ferritin-based VLP.
  • the protein complex is a protein-based VLP (including ferritin, E2p, 13-01 and 13-01 variants) as described in U.S. Pat. Pub. No.
  • the protein- based VLP may employ a variety of coupling techniques to attach an antigen to the VLP core, including but not limited to the SpyCatcher system described in, e.g., Escolano et al. Nature 570:468-473 (2019), He et al. Sci Adv. 7(12):eabfl591 (2021), and Tan et al. Nat. Commun. 12(1):542 (2021).
  • the protein-based VLP may be a lumazine synthase nanoparticle as described, e.g., in Geng et al. PLoS Pathog. 17(9):el009897 (2021).
  • the protein-based VLP maybe a ferritin nanoparticle as described, e.g., in Joyce et al. bioRxiv 2021.05.09.443331 and in U.S. Pat. Pub. No. US 2019/0330279 AL
  • the RBD or coronavirus S protein ectodomain, or antigenic fragments thereof are expressed as a fusion protein with the first multimerization domain.
  • the first multimerization domain and RBD or coronavirus S protein ectodomain are joined by a linker sequence.
  • the linker sequence comprises a foldon, wherein the foldon sequence is EKAAKAEEAARK (SEQ ID NO: 8).
  • the linker may comprise a Gly-Ser linker (i.e . a linker consisting of glycine and serine residues) of any suitable length.
  • the Gly-Ser linker may be 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more amino acids in length.
  • 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; Int’l 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 3.
  • 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: 9-13 and comprises an RBD or coronavirus S 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: 13-18 or 27.
  • the VLP comprises a fusion protein that has at least 75% identity to any one of SEQ ID NO: 9-13 and comprises an RBD or coronavirus S protein as disclosed herein; and a second component that has at least 75% identity to any one of SEQ ID NO: 13-18 or 27.
  • 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: 19 and comprises an RBD or coronavirus S 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: 20.
  • 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: 1-6.
  • amino acid sequence of the native or wild-type SARS-CoV-2 S protein, subunit 1 is:
  • the first component may comprise a receptor-binding domain of a coronavirus S protein.
  • the receptor-binding domain of a coronavirus S 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: 21-24.
  • 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: 1-6 and further comprises a signal peptide.
  • the signal peptide comprises the sequence of SEQ ID NO: 25.
  • the signal peptide comprises the sequence of SEQ ID NO: 26.
  • MGILPSPGMPALLSLV SLLS VLLMGCVA SEQ ID NO: 25
  • MGILPSPGMPALLSLVSLLSVLLMGCVAETGT SEQ ID NO: 26
  • polypeptides as described herein may have one of more amino acid substitutions from known variants of SARS-CoV2 (also called “variant strains of SARS-CoV2”). Such variant strains of SARS-CoV2 comprise mutations relative to the original strain of SARS-CoV2.
  • original strain as used herein refers to the Wuhan strain of SARS-CoV-2 identified in 2019- 2020.
  • the polypeptides may comprise 1, 2, 3, 4, 5, 6, 7, or all 8 positions relative to SEQ ID NO: 7 selected from the group consisting of L18F, T20N, P26S, deletion of residues 69-70, D80A, D138Y, R190S, D215G, R346K, K417N, K417T, G446S, L452R, Y453F, S477N, T478I, T478K, V483A, E484K, E484Q, S494P, N501Y, A570D, D614G, H655Y, G669S, Q677H, P681H, P681R, A701V, T716L.
  • the polypeptides may comprise one of the following naturally occurring mutations or combinations of mutations:
  • N501Y optionally further including 1, 2, 3, 4, or 5 of deletion of one or both of residues 69-70, E484K, A570D, D614G, P681H, and/or T716L (UK variant);
  • K417N/E484K/N501Y optionally further including 1, 2, 3, 4, or 5 ofL18F, D80A, D215G, D614G, and/or A701V (South African variant);
  • K417N or T/E484K/N501Y optionally further including 1, 2, 3, 4, or 5 of L18F, T20N, P26S, D138Y, R190S, D614G, and/or H655Y (Brazil variant);
  • V483A, D614G, H655Y, G669S France variant
  • 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 exemplary conservative amino acid substitutions is given in the table below.
  • Non-conservative amino acid substitution may be preferred, for example, when eradication of a flexible portion of the native coronavirus S protein secondary structure is desired, for example, by adding a cysteine residue (or vice versa).
  • “Non-conservative 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
  • a polar (hydrophilic) residue such as cysteine, glutamine, glutamic acid or lysine and/or a polar residue for a non-polar residue.
  • 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, steroid-responsive).
  • 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 SARS-CoV-2.
  • the virus-like particles of the disclosure induce neutralizing antibody titers roughly ten-fold higher than the prefusion-stabilized S ectodomain trimer despite a more than five-fold lower dose.
  • Antibodies elicited by the virus-like particles target multiple distinct epitopes, suggesting that they may not be easily susceptible to escape mutations, and exhibit a significantly lower binding eutralizing ratio than convalescent human sera, which may minimize the risk of vaccine-associated enhanced respiratory disease.
  • compositions/vaccines may further comprise (a) a lyoprotectant; (b) a surfactant; (c) a bulking agent; (d) a tonicity adjusting agent; (e) a stabilizer; (f) 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.
  • Exemplary 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.
  • Exemplary 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, 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.
  • Exemplary 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. benzonaphthyridines; 17. microparticles; 18. immunostimulatory polynucleotide (such as RNA or DNA; e.g., cpg-containing oligonucleotides).
  • pcpp polyphosphazene
  • Exemplary adjuvants that may be used in a pharmaceutical composition provided herein include, but are not limited to, 3M-052, Adju-PhosTM, AdjumerTM, albumin-heparin microparticles, Algal Glucan, Algammulin, Alum, Antigen Formulation, AS-2 adjuvant, ASOl, AS03, autologous dendritic cells, autologous PBMC, AvridineTM, B7-2, BAK, BAY R1005, Bupivacaine, Bupivacaine-HCl, BWZL, Calcitriol, Calcium Phosphate Gel, CCR5 peptides, CFA, Cholera holotoxin (CT) and Cholera toxin B subunit (CTB), Cholera toxin A 1 -subunit-Protein A D-fragment fusion protein, CpG, CPG-1018, CRL1005, Cytokine-containing Liposomes, D- Murapalmitine, DDA, DHEA
  • 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 or AS03), a TLR7 agonist (such as imidazoquinoline or imiquimod), or a combination thereof.
  • the adjuvant is a combination of an aluminum salt and CPG-1018.
  • Suitable aluminum salts include hydroxides (e.g. oxyhydroxides), phosphates (e.g. hydroxyphosphates, orthophosphates), (e.g. see chapters 8 & 9 of Vaccine Design. (1995) eds. Powell & Newman. ISBN: 030644867X.
  • the salts can take any suitable form (e.g. gel, crystalline, amorphous, etc.), with adsorption of antigen to the salt being an example.
  • 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.
  • a preferred range is between 0.3 and 1 mg/ml.
  • a maximum of 0.85mg/dose is preferred.
  • Aluminum hydroxide and aluminum phosphate adjuvants are suitable for use with the disclosure.
  • 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.
  • 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 — for example, 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 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, or less than 0.1% (w/v) of the pharmaceutical composition.
  • 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).
  • an adjuvant e.g., Alum
  • unit doses of the pharmaceutical composition described herein comprises about 5 pg to about 10 pg, about 10 pg to about 15 pg, about 15 pg to about 20 pg, about 20 pg to about 30 pg, about 30 pg to about 40 pg, about 40 pg to about 50 pg, about 50 pg to about 60 pg, about 60 pg to about 70 pg, about 70 pg to about 80 pg, about 80 pg to about 90 pg, about 90 pg to about 100 pg, about 100 pg to about 110 pg, about 110 pg to about 120 pg, about 120 pg to about 130 pg, about 130 pg to about 140 pg, about 140
  • the unit dosage comprises 2 pg, 5 pg, 10 pg, 15 pg, 25 pg, 50 pg, 100 pg, or 125 pg of the protein complex. In some embodiments, the unit dosage comprises 5 pg of the protein complex. In some embodiments, the unit dosage comprises 25 pg of the protein complex. In some embodiments, the unit dosage comprises 125 pg of the protein complex. In some embodiments, the unit dosage comprises 100 pg of the protein complex.
  • a unit dose of the pharmaceutical composition described herein wherein the unit dose comprises between about 25 pg and about 125 pg of the protein complex.
  • the unit dose of the pharmaceutical composition is between about 2 pg to about 125 pg, or between about 5 pg to about 125 g, or between about 15 pg to 125 pg, or between about 25 pg to about 125 pg, or between about 50 pg to about 125 pg, or between about 100 pg to about 125 pg of the 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 comprising the mutated coronavirus proteins described 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.
  • the disclosure provides methods to treat or limit development of a SARS- CoV-2 infection (e.g., infection with an original strain of SARS-CoV2 or infection with a variant strain of SARS-CoV2), 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 subject may be any suitable mammalian subject, including but not limited to a human subject.
  • Examples of variant strains of SARS-CoV2 have been detected around the world and include, without limitation: B.l.1.7 (UK), B.1.1.7+E484K (UK), B.1.351 (South Africa), P.l (Brazil), B.1.617.2 (India), B.1.525 (Nigeria), B.1.427/B.1.429 (USA), P.3 (Philippines), B.1.616 (France), B.l.617.1 (India), B.l.617.3 (India), B.1.621 (Colombia), A.23.1+E484K (UK), C.37 (Peru), B.1.351+P384L (South Africa), B.1.1.7+L452R (UK), B.1.1.7+S494P (UK), C.36+L452R (Egypt), AT.l (Russia), B.1.526 (USA), B.1.526.1 (USA), B.1.526.2 (USA), B.1.1.318, P.2 (Brazil), B.
  • the immunogenic composition is administered prophylactically to a subject that is not known to be infected but may be at risk of exposure to SARS-CoV-2.
  • limiting development 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 of SARS-CoV-2 in the subject; (b) generating neutralizing antibodies against SARS-CoV- 2 in the subject (b) limiting build-up of SARS-CoV-2 titer in the subject after exposure to SARS- CoV-2; and/or (c) limiting or preventing development of SARS-CoV-2 symptoms after infection.
  • the methods provided herein may be used to limit development of infection with an original strain of SARS-CoV2 and/or infection with a variant strain of SARS-CoV2.
  • Exemplary symptoms of SARS-CoV-2 infection include, but are not limited to, fever, fatigue, cough, shortness of breath, chest pressure and/or pain, loss or diminution of the sense of smell, loss or diminution of the sense of taste, and respiratory issues including but not limited to pneumonia, bronchitis, severe acute respiratory syndrome (SARS), and upper and lower respiratory tract infections.
  • SARS severe acute respiratory syndrome
  • the methods generate an immune response in a subject in the subject not known to be infected with SARS-CoV-2, wherein the immune response serves to limit development of infection and symptoms of a SARS-CoV-2 infection (e.g., infection with an original strain of SARS-CoV2 or infection with a variant strain of SARS-CoV2).
  • the immune response comprises generation of neutralizing antibodies and/or cell- based responses against SARS-CoV-2.
  • the immune response comprises generation of SARS-CoV-2 S protein or RBD antibody-specific responses with a mean geometric titer of 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 assay units.
  • the immune response comprises generation of SARS- CoV-2 S protein or RBD antibody-specific responses with a mean geometric titer of at least 1 x 10 5 .
  • the immune response comprises generation of antibodies against multiple antigenic epitopes.
  • an “effective amount” refers to an amount of the immunogenic composition that is effective for treating and/or limiting SARS-CoV-2 infection (e.g., infection with an original strain of SARS-CoV2 or infection with a variant strain of SARS-CoV2).
  • 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 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
  • a method of vaccinating a subject at risk of infection with SARS-CoV-2 comprising administering to the subject a pharmaceutical composition comprising an effective amount of the protein complex comprising a first component comprising three receptor-binding domain monomers of a coronavirus S protein and a first multimerization domain (e.g., a trimerization domain), and a second component comprising a second multimerization domain (e.g., a pentamerization domain); and one or more pharmaceutically acceptable diluents or excipients.
  • the pharmaceutical composition comprises an adjuvant.
  • the adjuvant is or comprises an oil.
  • the adjuvant is an oil-in-water (e.g., a squalene-in-water) emulsion.
  • the adjuvant is MF59®.
  • the adjuvant is an aluminum salt.
  • the adjuvant is CPG-1018.
  • the pharmaceutical composition comprises both an aluminum salt and CPG-1018.
  • the effective amount is 2 mg, 5 pg, 10 pg, 15 pg, 25 pg, 50 pg, 100 pg, or 125 pg of the protein complex.
  • the method comprises repeating the administering step.
  • the method comprises administering a booster vaccine.
  • the subject is previously vaccinated with a SARS-CoV-2 vaccine and/or previously infected with SARS-CoV-2.
  • the subject has completed a full course of vaccination for SARS-CoV-2.
  • the subject has completed a full course of vaccination for an original strain vaccine of SARS-CoV-2.
  • the subject has completed a partial course (e.g., has received one of two doses of a full course) of vaccination for an original strain of SARS-CoV-2.
  • the subject has received at least one dose of a vaccination for a variant strain of SARS-CoV-2 (e.g. a variant strain described herein).
  • the term “partial course” refers to a first administration of a series of two or more administrations constituting an art- recognized full course of vaccination.
  • the subject has received at least one dose of a vaccine comprising the receptor binding domain of a coronavirus S protein or a polynucleotide encoding the receptor binding domain of a coronavirus S protein.
  • the subject has received at least one dose of a vaccine comprising a coronavirus S protein or a polynucleotide encoding a coronavirus S protein.
  • the S protein is S2P.
  • the S protein is “HexaPro” — /. e. , comprises the amino acid substitutions F817P, A892P, A899P, and A942P relative to a reference sequence.
  • the method induces neutralizing antibody titers in the subject. In some embodiments, the method increases neutralizing antibody titers in the subject. In some embodiments, the method induces S protein-specific and RBD-specific IgG antibody titers in the subject. In some embodiments, the method induces cell mediated immunity (CD4 T cells, memory B cells, CD8 T cells) in the subject. In some embodiments, the method induces neutralizing antibody titers in the subject. In some embodiments, the method prevents infection with an original strain of SARS-CoV-2. In some embodiments, the method prevents infection with a variant strain of SARS-CoV-2. In some embodiments, the method reduces the severity of infection with an original strain of SARS-CoV-2. In some embodiments, the method reduces the severity of infection with a variant strain of SARS-CoV-2
  • the methods herein include vaccinating a subject at risk of infection with SARS-CoV-2, comprises administering the composition (e.g., a vaccine comprising a protein complex comprising a first component comprising a receptor-binding domain of a coronavirus S protein attached to a first multimerization domain, and a second component comprising a second multimerization domain; and one or more pharmaceutically acceptable diluents or excipients) to a subject that is at least 1 year old, 2 years old, 3 years old, 4 years old, 5 years old, 6 years old, 7 years old, 9 years old, 10 years old, 12 years old, 15 years old, 20 years old, 30 years old, 40 years old, 50 years old, 60 years old, 70 years old, 80 years old, or 90 years old.
  • the composition e.g., a vaccine comprising a protein complex comprising a first component comprising a receptor-binding domain of a coronavirus S protein attached to a first multimerization domain, and a second
  • the subject is an adult at least 18 years old. In some embodiments, the subject is an elderly adult that is at least 60 years old, or at least 70 years old, or at least 80 years old, or at least 90 years old. In some embodiments, the subject is a child from about 2 years old to about 18 years old, or from about 5 years old to about 18 years old, or from about 10 years old to about 18 years old. In some embodiments, the subject is an infant that is one year old, or 6 months old, or 3 months old. In some embodiments, the subject is a child under 5 years of age, or under 4 years of age, or under 3 years of age, or under 2 years of age, or under 1 year of age.
  • the subject is at least about 1 month old, about 2 months old, about 3 months old, about 4 months old, about 5 months old, about 6 months old, about 7 months old, about 8 months old, about 9 months old, about 10 months old, or about 11 months old. In some embodiments, the subject is at least about
  • a method of vaccinating a subject comprising administering to the subject (i) a pharmaceutical composition comprising an effective amount of the protein complex comprising a first component comprising a receptor-binding domain of a coronavirus S protein and a first multimerization domain (e.g., a trimerization domain), and a second component comprising a second multimerization domain (e.g., a pentamerization domain).
  • a pharmaceutical composition comprising an effective amount of the protein complex comprising a first component comprising a receptor-binding domain of a coronavirus S protein and a first multimerization domain (e.g., a trimerization domain), and a second component comprising a second multimerization domain (e.g., a pentamerization domain).
  • the subject has received at least one dose of a vaccination for SARS-CoV-
  • the subject has received at least one dose of a vaccination for SARS-CoV-2 no less than 3 months before the administration of the protein complex. In some embodiments the subject has received at least one dose of a vaccination for SARS-CoV-2 about 3 months to about 6 months before the administration of the protein complex. In some embodiments the subject has received at least one dose of a vaccination for SARS-CoV-2 3 months to 6 months before the administration of the protein complex. In some embodiments, the subject has received at least one dose of a vaccine comprising the receptor binding domain of a coronavirus S protein or a polynucleotide encoding the receptor binding domain of a coronavirus S protein.
  • the subject has received at least one dose of a vaccine comprising a coronavirus S protein or a polynucleotide encoding a coronavirus S protein.
  • the S protein is S2P or “HexaPro” — /. e. , comprises the amino acid substitutions F817P, A892P, A899P, and A942P relative to a reference sequence.
  • the vaccine is an mRNA-based vaccine, an adenoviral vector- based vaccine, a protein-subunit based vaccine, or an inactivated virus vaccine.
  • the subj ect has completed a full course of vaccination for an original strain of S ARS- CoV-2.
  • the subject has completed a partial course (e.g., has received one of two doses) of vaccination for an original strain of SARS-CoV-2.
  • the subject has previously been infected with SARS-CoV-2.
  • the subject has antibodies against SARS-CoV-2.
  • the subject has not previously been infected with SARS-CoV-2.
  • the method induces neutralizing antibody titers in the subject. In some embodiments, the method induced SARS-CoV-2 ancestral strain specific neutralizing antibody titers in the subject. In some embodiments, the method induces a serum SARS-CoV-2 binding antibody response in the subject. In some embodiments, the antibody response is to a SARS-CoV-2 RBD and/or a SARS-CoV-2 S protein. In some embodiments, the method prevents infection with an original strain of SARS-CoV-2 and/or a variant strain of SARS-CoV-2. In some embodiments, the method reduces the severity of infection with coronavirus.
  • 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 pg/kg to 0.5 pg /kg body weight, 0.5 pg/kg to 1 pg body weight, 1 pg/kg to 2 pg/kg body weight, 2 pg/kg to 3 pg/kg body weight, 3 pg/kg to 4 pg/kg body weight, 4 pg/kg to 5 pg/kg body weight, 5 pg/kg to 6 mg/kg body weight, 6 mg/kg to7 mg/kg body weight, 7 mg/kg to 8 mg/kg body weight, 8 mg/kg to 9 mg/kg body weight, 9 mg/kg to 10 mg/kg body weight, 10 mg/kg to 15 mg/kg body weight, 15 mg/kg to 20 mg/kg body weight, 20 mg/kg to 25 mg/kg body weight, 25 mg/kg to 30 mg/kg body weight, 30 mg/
  • 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 pg to about 100 pg, about 10 pg to about 150 pg, about 10 pg to about 200 pg, about 10 pg to about 250 pg, about 10 pg to about 300 pg, about 10 pg to about 350 pg, about 10 pg to about 400 pg, about 10 pg to about 450 pg, or about 10 pg to about 500 pg of the protein complex are administered.
  • about 25 pg to about 100 pg, about 25 pg to about 150 pg, about 25 pg to about 200 pg, about 25 pg to about 250 pg, about 25 pg to about 300 pg, about 25 pg to about 350 pg, about 25 pg to about 400 pg, about 25 pg to about 450 pg, or about 25 pg to about 500 pg of the protein complex are administered.
  • about 50 pg to about 100 pg, about 50 pg to about 150 pg, about 50 pg to about 200 pg, about 50 pg to about 250 pg, about 50 pg to about 300 pg, about 50 pg to about 350 pg, about 50 pg to about 400 pg, about 50 pg to about 450 pg, or about 50 pg to about 500 pg of the protein complex are administered.
  • about 5 mg to about 150 pg, about 10 pg to about 150 pg, about 25 pg to about 150 pg, about 50 pg to about 150 pg, about 75 pg to about 150 pg, about 100 pg to about 150 pg, or about 125 pg to about 150 pg of the protein complex are administered.
  • about 5 pg to about 125 pg, about 10 pg to about 125 pg, about 25 pg to about 125 pg, about 50 pg to about 125 pg, about 75 pg to about 125 pg, or about 100 pg to about 125 pg of the protein complex are administered.
  • about 5 pg to about 100 pg, about 10 pg to about 100 pg, about 25 pg to about 100 pg, about 50 pg to about 100 pg, or about 75 pg to about 100 pg of the protein complex are administered.
  • about 5 pg to about 75 pg, about 10 pg to about 75 pg, about 25 pg to about 75 pg, or about 50 pg to about 75 pg of the protein complex are administered.
  • about 5 pg to about 50 pg, about 10 pg to about 50 pg, or about 25 pg to about 50 pg of the protein complex are administered.
  • the dose amount described herein can be converted to molar amounts or adjusted, depending on the molecular mass of the protein complex, to deliver the same or similar molar amounts of the antigen (RBD).
  • the protein complexes of the disclosure generally have molecular masses of about 4 MDa (60 copies each of 50 kDa CompA-RBD and 17 kDa CompB).
  • the RBDs of the disclosure generally have molecular masses of about 23 kDa. Accordingly, 100 pg of a protein complex may be about 2.5 picomoles (pmol), and each 100 pg of protein complex may include about 34 pg of the RBD.
  • about 0.25 pmol to about 10 pmol, about 0.25 pmol to about 15 pmol, about 0.25 pmol to about 20 pmol, about 0.25 pmol to about 25 pmol, about 0.25 pmol to about 30 pmol, about 0.25 pmol to about 35 pmol, about 0.25 pmol to about 40 pmol, about 0.25 pmol to about 45 pmol, or about 0.25 pmol to about 50 pmol of the protein complex are administered.
  • about 0.5 pmol to about 10 pmol, about 0.5 pmol to about 15 pmol, about 0.5 pmol to about 20 pmol, about 0.5 pmol to about 25 pmol, about 0.5 pmol to about 30 pmol, about 0.5 pmol to about 35 pmol, about 0.5 pmol to about 40 pmol, about 0.5 pmol to about 45 pmol, or about 0.5 pmol to about 50 pmol of the protein complex are administered.
  • about 1 pmol to about 10 pmol, about 1 pmol to about 15 pmol, about 1 pmol to about 20 pmol, about 1 pmol to about 25 pmol, about 1 pmol to about 30 pmol, about 1 pmol to about 35 pmol, about 1 pmol to about 40 pmol, about 1 pmol to about 45 pmol, or about 1 pmol to about 50 pmol of the protein complex are administered.
  • about 2 pmol to about 10 pmol, about 2 pmol to about 15 pmol, about 2 pmol to about 20 pmol, about 2 pmol to about 25 pmol, about 2 pmol to about 30 pmol, about 2 pmol to about 35 pmol, about 2 pmol to about 40 pmol, about 2 pmol to about 45 pmol, or about 2 pmol to about 50 pmol of the protein complex are administered.
  • about 5 pmol to about 10 pmol, about 5 pmol to about 15 pmol, about 5 pmol to about 20 pmol, about 5 pmol to about 25 pmol, about 5 pmol to about 30 pmol, about 5 pmol to about 35 pmol, about 5 pmol to about 40 pmol, about 5 pmol to about 45 pmol, or about 5 pmol to about 50 pmol of the protein complex are administered.
  • about 0.5 pmol to about 15 pmol, about 1 pmol to about 15 pmol, about 2 pmol to about 15 pmol, about 5 pmol to about 15 pmol, about 7 pmol to about 15 pmol, about 10 pmol to about 15 pmol, or about 12 pmol to about 15 pmol of the protein complex are administered.
  • about 0.5 pmol to about 12 pmol, about 1 pmol to about 12 pmol, about 2 pmol to about 12 pmol, about 5 pmol to about 12 pmol, about 7 pmol to about 12 pmol, or about 10 pmol to about 12 pmol of the protein complex are administered.
  • about 0.5 pmol to about 10 pmol, about 1 pmol to about 10 pmol, about 2 pmol to about 10 pmol, about 5 pmol to about 10 pmol, or about 7 pmol to about 10 pmol of the protein complex are administered.
  • about 0.5 pmol to about 7 pmol, about 1 pmol to about 7 pmol, about 2 pmol to about 7 pmol, or about 5 pmol to about 7 pmol of the protein complex are administered.
  • about 0.5 pmol to about 5 pmol, about 1 pmol to about 5 pmol, or about 2 pmol to about 5 pmol of the protein complex are administered.
  • about 10 pg to about 100 pg, about 10 pg to about 150 pg, about 10 pg to about 200 pg, about 10 pg to about 250 pg, about 10 pg to about 300 pg, about 10 pg to about 350 pg, about 10 pg to about 400 pg, about 10 pg to about 450 pg, or about 10 pg to about 500 pg of the RBD are administered.
  • about 50 pg to about 100 pg, about 50 pg to about 150 pg, about 50 pg to about 200 pg, about 50 pg to about 250 pg, about 50 pg to about 300 pg, about 50 pg to about 350 pg, about 50 pg to about 400 pg, about 50 pg to about 450 pg, or about 50 pg to about 500 pg of the RBD are administered.
  • about 5 pg to about 150 pg, about 10 pg to about 150 pg, about 25 pg to about 150 pg, about 50 pg to about 150 pg, about 75 pg to about 150 pg, about 100 pg to about 150 pg, or about 125 pg to about 150 pg of the RBD are administered.
  • about 5 pg to about 125 pg, about 10 pg to about 125 pg, about 25 pg to about 125 pg, about 50 pg to about 125 pg, about 75 pg to about 125 pg, or about 100 pg to about 125 pg of the RBD are administered.
  • about 5 mg to about 100 pg, about 10 pg to about 100 pg, about 25 pg to about 100 pg, about 50 pg to about 100 pg, or about 75 pg to about 100 pg of the RBD are administered.
  • about 5 pg to about 75 pg, about 10 pg to about 75 pg, about 25 pg to about 75 pg, or about 50 pg to about 75 pg of the RBD are administered.
  • about 5 pg to about 50 pg, about 10 pg to about 50 pg, or about 25 pg to about 50 pg of the RBD are administered.
  • about 25 pg to about 125 of the protein complex is administered. In some embodiments, about 25 pg to about 100 of the protein complex is administered.
  • about 10 pg to about 125 of the protein complex is administered. In some embodiments, about 10 pg to about 100 of the protein complex is administered.
  • about 25 pg to about 125 of the protein complex is administered without an adjuvant. In some embodiments, about 25 pg to about 100 of the protein complex is administered without an adjuvant.
  • about 10 pg to about 125 of the protein complex is administered without an adjuvant. In some embodiments, about 10 pg to about 100 of the protein complex is administered without an adjuvant.
  • 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.
  • 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.
  • 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. For example, 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.
  • the first boost dose is administered at day 0, and the second boost dose is administered at day 28.
  • the first boost dose is administered at day 0, and the second boost dose is administered at 3 months
  • a method comprises administering a first dose and a second dose of the pharmaceutical 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 protein complexes and pharmaceutical compositions of the disclosure may also be used for heterologous prime -boost vaccination.
  • 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.
  • Suitable vaccines for use as primary vaccines or as heterologous boost vaccines may include those marketed for use in humans by Modema®, Pfizer®/BioNTech®, AstraZeneca®, Johnson & Johnson®, Novavax®, Sanofi®, SK Biosciences®, Medicago®, and Bavarian Nordic®.
  • the protein complexes and pharmaceutical compositions described herein may be used in heterologous vaccination strategies with these and other vaccines for SARS-CoV-2.
  • the administering comprises
  • a prime dose to the subject of a protein ⁇ e.g., a subunit vaccine
  • DNA, mRNA, inactivated virus, or adenoviral vector vaccine wherein the protein, DNA, mRNA, inactivated virus, or adenoviral vector vaccine comprising or encoding a coronavirus S protein or antigenic fragment thereof;
  • the administering comprises
  • a protein e.g ., a subunit vaccine
  • DNA, mRNA, inactivated virus, or adenoviral vector vaccine wherein the protein DNA, mRNA, inactivated virus, or adenoviral vector vaccine comprising or encoding a coronavirus S protein or antigenic fragment thereof.
  • any suitable protein e.g., a subunit vaccine
  • DNA, mRNA, inactivated virus, adenoviral vector vaccine, or protein-based vaccine may be used in conjunction with the immunogenic compositions of the present disclosure, including but not limited to vaccines to be developed as well as those available from Moderna®, Pfizer®/BioNTech®, AstraZeneca®, Johnson & Johnson®, Novavax®, Sanofi®, SK Biosciences®, Medicago®, and Bavarian Nordic®, etc.
  • the administering comprises
  • a boost dose to the subject of a protein ⁇ e.g., a subunit vaccine), DNA, mRNA, or adenoviral vector vaccine which is authorized for use to limiting SARS-CoV2 infection ⁇ e.g., any suitable DNA, mRNA, inactivated virus, adenoviral vector, or protein -based vaccine, including those available from Moderna®, Pfizer®/BioNTech®, AstraZeneca®,
  • the administering comprises
  • a protein e.g ., a subunit vaccine
  • DNA, mRNA, inactivated virus, or adenoviral vector vaccine which is authorized for use to limiting SARS-CoV2 infection (e.g., any suitable DNA, mRNA, inactivated virus, adenoviral vector, or protein-based vaccine); and
  • the subject is infected with a severe acute respiratory (SARS) virus, including but not limited to SARS-CoV-2, wherein the administering elicits an immune response against the SARS virus in the subject that treats a SARS virus infection in the subject.
  • SARS severe acute respiratory
  • the immunogenic compositions are administered to a subject that has already been infected with SARS-CoV-2, and/or who is suffering from symptoms (as described above) indicating that the subject is likely to have been infected with SARS-CoV-2.
  • the administering comprises
  • a protein e.g., a subunit vaccine
  • DNA, mRNA, inactivated virus, or adenoviral vector vaccine which is authorized for use to limiting SARS-CoV2 infection (e.g., any suitable DNA, mRNA, inactivated virus, adenoviral vector, or protein -based vaccine).
  • the subject is infected with a severe acute respiratory (SARS) virus, including but not limited to SARS-CoV-2, wherein the administering elicits an immune response against the SARS virus in the subject that treats a SARS virus infection in the subject.
  • SARS severe acute respiratory
  • the immunogenic compositions are administered to a subject that has already been infected with SARS-CoV-2, and/or who is suffering from symptoms (as described above) indicating that the subject is likely to have been infected with SARS-CoV-2.
  • the subject has received one or more doses of a DNA, inactivated virus, mRNA, adenoviral vector, or protein-based vaccine which is authorized for use to limit SARS-CoV2 infection (e.g., any suitable DNA, mRNA, inactivated virus, adenoviral vector, or protein-based vaccine).
  • a DNA, inactivated virus, mRNA, adenoviral vector, or protein-based vaccine which is authorized for use to limit SARS-CoV2 infection (e.g., any suitable DNA, mRNA, inactivated virus, adenoviral vector, or protein-based vaccine).
  • the subject has received two doses of a DNA, mRNA, inactivated virus, adenoviral vector, or protein-based vaccine which is authorized for use to limiting SARS-CoV2 infection (e.g., any suitable DNA, mRNA, inactivated virus, adenoviral vector vaccine, or protein-based vaccine).
  • the subject has received a full course of a DNA, mRNA, inactivated virus, adenoviral vector, or protein-based vaccine which is authorized for use to limiting SARS-CoV2 infection (e.g., any suitable DNA, mRNA, inactivated virus, adenoviral vector, or protein-based vaccine).
  • the DNA, mRNA, inactivated virus, adenoviral vector, or protein- based vaccine is a vaccine against an original strain of SARS-CoV2. In some embodiments, the DNA, mRNA, inactivated virus, adenoviral vector, or protein-based vaccine is a vaccine against a variant strain of SARS-CoV2.
  • a subject has received at least one dose of a vaccine comprising the receptor binding domain of a coronavirus S protein or a polynucleotide encoding the receptor binding domain of a coronavirus S protein prior to receiving a dose or boost of any embodiment or combination disclosed herein.
  • a subject has received at least one dose of a vaccine comprising a coronavirus S protein or a polynucleotide encoding a coronavirus S protein prior to receiving a dose or boost of any embodiment or combination disclosed herein.
  • the S protein is S2P.
  • the S protein is HexaPro.
  • the one or more doses of the DNA, mRNA, inactivated virus, adenoviral vector, or protein-based vaccine which is authorized for use to limit SARS-CoV2 infection may be administered to a subject treated in accordance with the methods described herein about 1-2 weeks, about 2-3 weeks, about 3-4 weeks, about 4-5 weeks, about 5-6 weeks, about 6-7 weeks, about 7-8 weeks, about 8-9 weeks, about 9-10 weeks, about 10-11 weeks, about 11-12 weeks, about 3-4 months, about 4-5 months, about 5-6 months, about 6-7 months, about 7-8 months, about 8-9 months about 9-10 months, about 10-11 months, about 11-12 months, about 12-15 months, about 15-18 months, about 18-21 months, about 21-24 months, about 2-3 years, about 3-4 years, or about 4-5 years before the administration of an immunogenic composition provided herein.
  • the subject is a vaccination-nai ' ve subject. In some embodiments, the subject is naive to vaccinations to limit infection with a coronavirus. In some embodiments, the subject is naive to vaccinations to limit infection with SARS-CoV2.
  • the vaccine is a vaccination against an original strain of SARS- CoV2. In some embodiments, the vaccine is a vaccination against a variant strain of SARS-CoV2.
  • the term “authorized for use” in the context of a vaccine means a vaccine has been approved for use in humans by a regulatory authority (e.g ., the U.S. Food and Drug Administration, the European Medicines Agency, the Chinese National Medical Products Administration, the United Kingdom Medicines and Healthcare products Regulatory Agency, the Japanese Pharmaceutical Food and Medical Devices Agency, or the Russian Ministry of Health).
  • a regulatory authority e.g ., the U.S. Food and Drug Administration, the European Medicines Agency, the Chinese National Medical Products Administration, the United Kingdom Medicines and Healthcare products Regulatory Agency, the Japanese Pharmaceutical Food and Medical Devices Agency, or the Russian Ministry of Health.
  • Authorized for use can include emergency use authorization.
  • a subject treated in accordance with the methods described herein has previously been infected with SARS-CoV-2.
  • SARS-CoV-2 infection may be diagnosed using any PCR-based test or antigen-based test known in the art.
  • the subject has antibodies against SARS-CoV2 (e.g., an original strain or a variant strain) prior to the administering step.
  • Anti-S ARS-CoV2 antibodies may be detected using any serological test known in the art, including, for example, a test for IgM/IgG to the nucleocapsid protein, or a test for neutralizing antibodies against SARS-CoV2.
  • a subject treated in accordance with the methods described herein has not previously been infected with SARS-CoV2.
  • the subject does not have antibodies against SARS-CoV2 prior to the administering step.
  • “treat” or “treating” includes, but is not limited to accomplishing one or more of the following: (a) reducing SARS-CoV-2 titer in the subject; (b) limiting any increase of SARS-CoV-2 titer in the subject; (c) reducing the severity of SARS-CoV-2 symptoms; (d) limiting or preventing development of SARS-CoV-2 symptoms after infection; (e) inhibiting worsening of SARS-CoV-2 symptoms; (f) limiting or preventing recurrence of SARS-CoV-2 symptoms in subjects that were previously symptomatic for SARS-CoV-2 infection; and/or (e) survival.
  • full course refers the one or more administrations (e.g ., injections) of a vaccine or combination of vaccines as considered in the art to provide the desired level of protection against disease, such as a course of administration approved by a regulatory agency.
  • a full course may be a single administration.
  • nucleic acid- based vaccines e.g. mRNA-based vaccines
  • a full course is generally two administrations spaced apart by about one month.
  • a full course of vaccination may include two, three, four, or more administrations.
  • compositions and method described herein may be employed in subjects who have received one, two, three, four, or more administrations of a prior vaccine or vaccines.
  • a subject is up to date COVID-19 vaccines when they have received all doses in the primary series and all boosters recommended, when eligible (fully vaccinated subject).
  • a method of treatment described herein may further comprise the administration of a second vaccination to the subject.
  • the second vaccination is administered concurrently with the SARS-CoV-2 vaccination.
  • a method of vaccinating a subject comprising administering to the subject (i) a pharmaceutical composition comprising an effective amount of a SARS-CoV-2 vaccine (for example, the protein complex comprising a first component comprising a receptor-binding domain of a coronavirus S protein and trimerization domain, and a second component comprising a pentamerization domain).
  • a SARS-CoV-2 vaccine for example, the protein complex comprising a first component comprising a receptor-binding domain of a coronavirus S protein and trimerization domain, and a second component comprising a pentamerization domain.
  • the subject has received at least one dose of a vaccination for SARS- CoV-2 no less than 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 15 months, 18 months, 21 months, or 24 months before the administration of the combination of the SARS-CoV-2 vaccine and the second vaccine.
  • the subject has received at least one does of a vaccine comprising the receptor binding domain of a coronavirus S protein or a polynucleotide encoding the receptor binding domain of a coronavirus S protein, or a vaccine comprising a coronavirus S protein.
  • the S protein may be, for example, S2P or HexaPro.
  • the vaccine may be any approved vaccine for an original or a variant strain of SARS-CoV-2.
  • the vaccine may be an mRNA-based vaccine, an adenoviral vector-based vaccine, a protein-based vaccine, or an inactivated virus vaccine.
  • the subject may have received at least one dose, at least two doses, or at least three doses of the SARS- CoV-2 vaccine.
  • the subject has completed a full course of vaccination for an original or a variant SARS-CoV-2 strain.
  • the subject has been previously infected with SARS-CoV-2.
  • protein complexes and pharmaceutical compositions of the disclosure may be indicated for use in one or more of the following:
  • 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.
  • the kit comprises the composition comprising the protein complex and, separately, a composition comprising an adjuvant, such that the two compositions may be mixed prior to administration, or alternatively coadministered.
  • IVX-411 is a SARS-CoV-2 virus-like particle (VLP) vaccine targeting the original strain and incorporating the ACE2 receptor binding domain (RBD) from the SARS-CoV-2 S protein, a conserved antigen that induces neutralizing antibodies to several known epitopes, including those that prevent viral entry.
  • the RBD protein is genetically fused to Component A and manufactured in mammalian cells. Component A-RBD is then combined with the same Component B used for our other programs to make the fully assembled VLPs, each of which incorporate 60 copies of the monomeric RBD antigen.
  • the assembled protein complex is illustrated in FIG. 1.
  • IVX-411 may be used in the clinic in both aqueous (non-adjuvanted) and adjuvanted formulations.
  • IVX-411 was tested in mice, rats, and nonhuman primates. Intramuscular injection of these VLPs induced strong neutralizing antibody responses, with titers observable after a single priming dose and significantly increased titers observable after a boosting dose.
  • the immunogenicity generated in mice after vaccination with closely related precursor molecules, and formulated with an oil-in-water adjuvant has been shown to be durable, with neutralizing antibody titers remaining as high 20-24 weeks following the boosting dose as they were two weeks post-boost.
  • preclinical nonhuman primate data on a closely related precursor candidate assessed with several different adjuvant formulations have shown induction of robust neutralizing antibody titers well in excess of titers seen in human convalescent sera, and protection from viral challenge.
  • a GLP toxicology repeat intramuscular dose study was completed in rats. The study evaluated both injection site and systemic reactions to IVX-411, including non-adjuvanted and adjuvanted formulations. No test article-related effects were seen following administration of IVX- 411 on mortality, clinical observations, ophthalmic observations, body weights, food consumption, or body temperature. No observed effects were considered adverse, and all observed effects were either partially or fully reversed 4 weeks following the last administration.
  • a Phase 1/2 trial is designed to evaluate the safety and immunogenicity of IVX-411 in primary and booster vaccinations.
  • the clinical trial design is summarized in FIG. 2.
  • Part 1 was a Phase 1 assessment of primary vaccination with IVX-411 in adults 18-69 years of age not previously exposed to SARS-CoV-2 (seronegative)
  • Part 2 was a Phase 2 assessment of IVX-411 booster vaccination in adults previously exposed through SARS-CoV-2 vaccination (seropositive).
  • IVX-411 was administered as two doses, either unadjuvanted or formulated with an oil-in-water adjuvant, administered 28-days apart.
  • the Phase 1/2 trial was a randomized, placebo-controlled observer-blind dose-escalation study for safety and immunogenicity of two intramuscular (IM) doses of IVX-411.
  • IM intramuscular
  • six formulations of IVX-411 were tested including three dose levels each to be tested with and without Seqirus’s proprietary adjuvant MF59®.
  • the candidate vaccine IVX-411 incorporates the angiotensin-converting enzyme 2 (ACE2) RBD from the SARS-CoV-2 spike (S) glycoprotein, a domain shown to be responsible for the majority (-90%) of the nAbs against the virus found in human convalescent sera. There are two components that are assembled to form the VLP DS.
  • ACE2 angiotensin-converting enzyme 2
  • S SARS-CoV-2 spike glycoprotein
  • the antigenic component (CompA-RBD-01) a structural component (CompB-01), when combined, self-assemble into an icosahedral VLP drug substance (DS).
  • the IVX-411 DS is a VLP made of 20 copies of the CompA-RBD-01 DSI (displaying 60 copies of the RBD as 20 sets of 3 RBD antigens) and 12 copies of CompB-01 DSI.
  • the selected adjuvant, MF59® (MF59C.1®; Seqirus, Inc), is an oil-in-water emulsion with a squalene internal oil phase and a citric acid- sodium citrate buffer external aqueous phase.
  • IVX-41 la is an aqueous buffer formulation of IVX-411 DS filled in single use vials for IM use.
  • IVX-41 la DP is supplied in single-use 2.0 mL vials at a concentration of 500 pg/mL at a 0.5 mL fill volume.
  • IVX-41 Id is a single-dose liquid formulation of IVX-41 la that has been mixed with the MF59® adjuvant.
  • MF59® is an oil-in-water emulsion with a squalene internal oil phase and a citric acid- sodium citrate buffer external aqueous phase.
  • Part 1 first-in-human (FIH), Phi
  • Part 2 Booster, Ph2
  • Both parts evaluated the safety and immunogenicity of two doses of IVX-411 vaccine administered 28 days apart, with or without MF59 adjuvant, in comparison with two doses of placebo.
  • Part 1 of the Phase 1/2 (Phl/2) study was a randomized, placebo-controlled observer-blind dose-escalation study for safety and immunogenicity of two intramuscular (IM) doses of IVX-411 administered 28-days apart (Day 0 and Day 28).
  • IM intramuscular
  • SARS-CoV-2 NAb titers by pseudovirion NAb assay
  • S-specific and RBD-specific IgG titers by enzyme-linked immunoassay (ELISA); and the ratios of fold-increase in RBD-specific IgG (multiplex assay) titers over fold-increase in SARS-CoV-2 NAb (live-virus assay) titers.
  • Immunogenicity assays used are listed in Table 2. Table 2
  • Part 2 was a Phase 2 assessment of booster vaccination with IVX-411 in 84 healthy adults who have been previously vaccinated with a licensed vaccine against SARS-CoV-2. The study determined whether an adjuvant was required in the formulation to enhance immune responses to IVX-411.
  • the selected adjuvant, MF59® was an oil-in-water emulsion. The clinical trial design is summarized in FIG. 4.
  • Example 4 Immunogenicity and Safety of a Protein-based Virus-Like Particle (VLP) SARS-CoV-2 Vaccine in Adults: a Phase 1/2 Study
  • Immunogenicity data showed immunogenicity in primary and booster vaccination (Neutralizing and IgG antibody titers exceed those of placebo recipients at Day 49 for WT, Clear evidence of adjuvant effect with more limited dose effect, with high rates of seroconversion, in primary regimen, Heterologous boosting after mRNA and adeno primary vaccination with up to 5-fold rise from baseline for WT, Immune responses seen across all variants of concern (beta, delta, omicron) in primary and booster context).
  • a pharmaceutical composition comprising a protein complex comprising a first component comprising a receptor-binding domain of a coronavirus S protein attached to a first multimerization domain, and optionally a second component comprising a second multimerization domain; and optionally one or more pharmaceutically acceptable diluents or excipients.
  • compositions of any one of embodiments 1 to 11, wherein 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 the group consisting of SEQ ID NOS: 14-17, 20 or 27.
  • 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: 14.
  • a method of vaccinating a subject at risk of infection with SARS-CoV-2 comprising administering to the subject a pharmaceutical composition comprising an effective amount of a protein complex comprising a first component comprising a receptor-binding domain of a coronavirus S protein attached to a first multimerization domain, and a second component comprising a second multimerization domain; and one or more pharmaceutically acceptable diluents or excipients.
  • a method of boosting an immune response to a prior vaccination for SARS-CoV-2 comprising administering to a subject previously vaccinated for SARS-CoV-2 a pharmaceutical composition comprising an effective amount of a protein complex comprising a first component comprising a receptor-binding domain of a coronavirus S protein attached to a first multimerization domain, and optionally a second component comprising a second multimerization domain.
  • a method of safely and effectively immunizing a subject for SARS-CoV-2 comprising administering to a subject previously vaccinated for SARS-CoV-2 a pharmaceutical composition comprising an effective amount of a protein complex comprising a first component comprising a receptor-binding domain of a coronavirus S protein attached to a first multimerization domain, and optionally a second component comprising a second multimerization domain.
  • 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: 14.
  • prime vaccine is an mRNA-based vaccine, an adenoviral vector-based vaccine, a protein— based vaccine, or an inactivated virus vaccine.

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