WO2020065350A1 - Vaccins - Google Patents

Vaccins Download PDF

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Publication number
WO2020065350A1
WO2020065350A1 PCT/GB2019/052748 GB2019052748W WO2020065350A1 WO 2020065350 A1 WO2020065350 A1 WO 2020065350A1 GB 2019052748 W GB2019052748 W GB 2019052748W WO 2020065350 A1 WO2020065350 A1 WO 2020065350A1
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seq
polypeptide
amino acid
acid sequence
nucleic acid
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PCT/GB2019/052748
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English (en)
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Jonathan Luke Heeney
Simon Frost
Ralf Wagner
Benedikt ASBACH
Rebecca KINSLEY
Edward Wright
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The Chancellor, Masters And Scholars Of The University Of Cambridge
University Of Westminster
Universitat Regensburg
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Application filed by The Chancellor, Masters And Scholars Of The University Of Cambridge, University Of Westminster, Universitat Regensburg filed Critical The Chancellor, Masters And Scholars Of The University Of Cambridge
Publication of WO2020065350A1 publication Critical patent/WO2020065350A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/12—Viral antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/70—Multivalent vaccine
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • 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
    • C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
    • C12N2710/00011—Details
    • C12N2710/24011—Poxviridae
    • C12N2710/24111—Orthopoxvirus, e.g. vaccinia virus, variola
    • C12N2710/24141—Use of virus, viral particle or viral elements as a vector
    • C12N2710/24143—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/10011—Arenaviridae
    • C12N2760/10022—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/10011—Arenaviridae
    • C12N2760/10034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/14011—Filoviridae
    • C12N2760/14022—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/14011—Filoviridae
    • C12N2760/14034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/14011—Filoviridae
    • C12N2760/14111—Ebolavirus, e.g. Zaire ebolavirus
    • C12N2760/14122—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/14011—Filoviridae
    • C12N2760/14111—Ebolavirus, e.g. Zaire ebolavirus
    • C12N2760/14134—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/14011—Filoviridae
    • C12N2760/14211—Marburgvirus, e.g. lake Victoria marburgvirus
    • C12N2760/14222—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/14011—Filoviridae
    • C12N2760/14211—Marburgvirus, e.g. lake Victoria marburgvirus
    • C12N2760/14234—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein

Definitions

  • This invention relates to nucleic acid molecules, polypeptides, vectors, cells, fusion proteins, pharmaceutical compositions, and their use as vaccines against emerging and re- emerging viruses, especially Filoviruses or Arenaviruses.
  • the fundamental principal of a vaccine is to prepare the immune system for an encounter with a pathogen.
  • a vaccine triggers the immune system to produce antibodies and T-cell responses, which help to combat infection.
  • Historically once a pathogen was isolated and grown, it was either mass produced and killed or attenuated, and used as a vaccine. Later recombinant genes from isolated pathogens were used to generate recombinant proteins that were mixed with adjuvants to stimulate immune responses.
  • pathogen genes were cloned into vector systems (attenuated bacteria or viruses) to express and deliver the antigen in vivo. All of these strategies are dependent on pathogens isolated from past outbreaks to prevent future ones. For pathogens which do not change significantly, or slowly, this conventional technology is effective. However, some pathogens, are prone to mutating and antibodies do not always recognise different strains of the pathogen. New emerging and re-emerging pathogens often hide or disguise their vulnerable antigens from the immune system.
  • RNA viruses are a virus that has RNA as its genetic material. This nucleic acid is usually single-stranded RNA (ssRNA) but may be double-stranded RNA (dsRNA). RNA viruses generally have very high mutation rates compared to DNA viruses, because viral RNA polymerases lack the proofreading ability of DNA polymerases. This is one reason why it is difficult to make effective vaccines to prevent diseases caused by RNA viruses.
  • RNA viruses In most cases, current vaccine candidates against RNA viruses are limited by the viral strain used as the vaccine insert, which is often chosen based on availability of a wild-type strain rather than by informed design.
  • Technical challenges for developing vaccines for enveloped RNA viruses include: i) viral variation of wild-type field isolate glycoproteins (GPs) provide limited breadth of protection as vaccine antigens; ii) selection of vaccine antigens expressed by the vaccine inserts is highly empirical; immunogen selection is a slow, trial and error process; iii) in an evolving or unanticipated viral epidemic, developing new vaccine candidates is time-consuming and can delay vaccine deployment.
  • Notable human diseases caused by RNA viruses include viral hemorrhagic fevers (VHFs), a group of illnesses that are caused by several distinct families of viruses.
  • VHFs viral hemorrhagic fevers
  • VHF viral hemorrhagic fever
  • a severe multisystem syndrome i.e. multiple organ systems in the body are affected. Characteristically, the overall vascular system is damaged, and the body’s ability to regulate itself is impaired. These symptoms are often accompanied by hemorrhage (bleeding), although the bleeding is itself rarely life- threatening. While some types of hemorrhagic fever viruses can cause relatively mild illnesses, many of the viruses cause severe, life-threatening disease. VHFs are caused by viruses of at least five distinct families: Arenaviridae, Bunyaviridae, FHoviridae, Flaviviridae, and Paramyxo viridae.
  • the viruses of these families are all RNA viruses, and are all covered, or enveloped, in a fatty (lipid) coating.
  • the survival of VHFs is dependent on an animal or insect host (the natural reservoir).
  • the viruses are geographically restricted to the areas where their host species live, and humans are infected when they come into contact with infected hosts. With some of the viruses, after transmission from the host, humans can transmit the virus to one another. Human cases or outbreaks of hemorrhagic fevers caused by these viruses occur sporadically and irregularly. The occurrence of outbreaks cannot be easily predicted. With a few exceptions, there is no cure or established drug treatment for VHFs.
  • VHFs caused by Arenaviruses and Filoviruses together cover a wide geographic region ranging from Western through to Central Africa and threaten adjacent regions where infected animal reservoirs may migrate but where human disease has not yet been reported.
  • Filoviruses encode their genome in the form of single-stranded negative-sense RNA.
  • Two members of the family that are commonly known are Ebola virus and Marburg virus. Ebola is an emerging and re-emerging RNA viral disease.
  • Aseptic meningitis a severe human disease that causes inflammation covering the brain and spinal cord, can arise from the Lymphocytic choriomeningitis virus (LCMV) infection.
  • Hemorrhagic fever syndromes are derived from infections such as Guanarito virus (GTOV), Junin virus (JUNV), Lassa virus (LASV), Lujo virus (LUJV), Machupo virus (MACV), Sabia virus (SABV), or Whitewater Arroyo virus (WWAV).
  • Lassa Fever virus (LASV), Ebola (EBOV) and Marburg (MARV) viruses are the most important haemorrhagic fevers in West and Central Africa. Lassa fever is endemic to Western Africa with estimates ranging between 300,000 to a million infections, with 5,000 deaths per year. Lassa Fever virus (LASV), Ebola (EBOV) and Marburg (MARV) viruses are all containment level 4 pathogens with high human morbidity and mortality for which there are no established cures, and currently there are no licensed vaccines for infections caused by these viruses. There is a need, therefore, to provide effective vaccines that induce a broadly neutralising immune response to protect against emerging and re-emerging diseases, especially those caused by viruses such as RNA viruses, including VHFs.
  • the applicant has identified nucleic acid molecules encoding amino acid sequences that, when expressed in a subject, induce broad neutralizing immune responses with broad neutralisation profiles against the Ebolavirus genus (Zaire, Sudan, Bundibugyo), additionally targeting the more distant filovirus, Marburg virus.
  • an isolated nucleic acid molecule comprising a nucleic acid sequence that is: i) at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical with SEQ ID NO:1 , or identical with SEQ ID NO:1 ; ii) at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical with SEQ ID NO:3, or identical with SEQ ID NO:3; iii) at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical with SEQ ID NO:5, or identical with SEQ ID NO:5; iv) at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical with SEQ ID NO:
  • an isolated nucleic acid molecule comprising a nucleic acid sequence that is: i) at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical with SEQ ID NO:14, or identical with SEQ ID NO:14; ii) at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical with SEQ ID NO:16, or identical with SEQ ID NO:16; iii) at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical with SEQ ID NO:18, or identical with SEQ ID NO. ⁇ 8; iv) at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical with SEQ ID NO:20, or identical with SEQ ID NO:20; v) at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or
  • an isolated polypeptide comprising an amino acid sequence that is: i) at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical with an amino acid sequence encoded by SEQ ID NO:1 , or identical with the amino acid sequence encoded by SEQ ID NO:1 ; ii) at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical with an amino acid sequence encoded by SEQ ID NO:3, or identical with the amino acid sequence encoded by SEQ ID NO:3; iit) at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical with an amino acid sequence encoded by SEQ ID NO:5, or identical with the amino acid sequence encoded by SEQ ID NO:5; iv) at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical with an amino acid sequence
  • an isolated polypeptide comprising an amino acid sequence that is: i) at least 95%, 96%, 97%, 98%, or 99% identical with SEQ ID NO:2, or identical with SEQ ID NO:2; ii) at least 95%, 96%, 97%, 98%, or 99% identical with SEQ ID NO:4, or identical with SEQ ID NO:4; or iii) at least 95%, 96%, 97%, 98%, or 99% identical with SEQ ID NO:6, or identical with SEQ ID NO:6; iv) at least 95%, 96%, 97%, 98%, or 99% identical with SEQ ID NO:8, or identical with SEQ ID NO:8; v) at least 95%, 96%, 97%, 98%, or 99% identical with SEQ ID NQ:10, or identical with SEQ ID NQ:10; or vi) at least 95%, 96%, 97%, 98%, or 99% identical with SEQ ID NO:12, or identical with SEQ ID NO:2; ii
  • an isolated polypeptide comprising an amino acid sequence that is: i) at least 95%, 96%, 97%, 98%, or 99% identical with SEQ ID NO:13, or identical with SEQ ID NO:13; ii) at least 95%, 96%, 97%, 98%, or 99% identical with SEQ ID NO:15, or identical with SEQ ID NO:15; iii) at least 95%, 96%, 97%, 98%, or 99% identical with SEQ ID NO:17, or identical with SEQ ID NO:17; iv) at least 95%, 96%, 97%, 98%, or 99% identical with SEQ ID NO:19, or identical with SEQ ID NO:19; v) at least 95%, 96%, 97%, 98%, or 99% identical with SEQ ID NO:21 , or identical with SEQ ID NO:21 ; vi) at least 95%, 96%, 97%, 98%, or 99% identical with SEQ ID NO:23, or identical with
  • narrowly neutralizing immune response is used herein to mean an immune response elicited in a subject that is sufficient to inhibit (i.e. reduce), neutralize or prevent infection, and/or progress of infection, of at least two different subtypes or species of a pathogen, for example at least two different subtypes or species of a virus, at least two different subtypes or species of a bacterium, or at least two different subtypes or species of a fungus.
  • a broadly neutralizing immune response is sufficient to inhibit, neutralize or prevent infection, and/or progress of infection, of most or all different subtypes or species of a pathogen, for example most or all different subtypes or species of a virus, most or all different subtypes or species of a bacterium, or most or all different subtypes or species of a fungus.
  • a broadly neutralizing immune response is sufficient to inhibit, neutralize or prevent infection, and/or progress of infection, of members of at least two different types of a pathogen (for example a virus, bacterium, or fungus) within the same family.
  • a broadly neutralizing immune response is sufficient to inhibit, neutralize or prevent infection, and/or progress of infection, of members of at least two different genera of a pathogen (for example a virus, bacterium, or fungus) within the same family.
  • the immune response may be humoral and/or a cellular immune response.
  • a cellular immune response is a response of a cell of the immune system, such as a B-cell, T-cell, macrophage or polymorphonucleocyte, to a stimulus such as an antigen or vaccine.
  • An immune response can include any cell of the body involved in a host defence response, including for example, an epithelial cell that secretes an interferon or a cytokine.
  • An immune response includes, but is not limited to, an innate immune response or inflammation.
  • a polypeptide of the invention induces a protective immune response.
  • a protective immune response refers to an immune response that protects a subject from infection or disease (i.e. prevents infection or prevents the development of disease associated with infection).
  • Methods of measuring immune responses include, for example, measuring proliferation and/or activity of lymphocytes (such as B or T cells), secretion of cytokines or chemokines, inflammation, or antibody production.
  • a polypeptide of the invention is able to induce the production of antibodies and/or a T-cell response in a human or non-human animal to which the polypeptide has been administered (either as a polypeptide or, for example, expressed from an administered nucleic acid expression vector).
  • sequence identity is frequently measured in terms of percentage identity (or similarity or homology); the higher the percentage, the more similar the two sequences are.
  • Homologs or variants of a given gene or protein will possess a relatively high degree of sequence identity when aligned using standard methods. Methods of alignment of sequences for comparison are well known in the art. Various programs and alignment algorithms are described in: Smith and Waterman, Adv. Appl. Math. 2:482, 1981 ; Needleman and Wunsch, J. Mol. Biol.
  • Biotechnology Information (NCBI, Bethesda, MD) and on the Internet, for use in connection with the sequence analysis programs blastp, blastn, blastx, tblastn and tblastx.
  • Sequence identity between nucleic acid sequences, or between amino acid sequences can be determined by comparing an alignment of the sequences. When an equivalent position in the compared sequences is occupied by the same nucleotide, or amino acid, then the molecules are identical at that position. Scoring an alignment as a percentage of identity is a function of the number of identical nucleotides or amino acids at positions shared by the compared sequences. When comparing sequences, optimal alignments may require gaps to be introduced into one or more of the sequences to take into consideration possible insertions and deletions in the sequences.
  • Sequence comparison methods may employ gap penalties so that, for the same number of identical molecules in sequences being compared, a sequence alignment with as few gaps as possible, reflecting higher relatedness between the two compared sequences, will achieve a higher score than one with many gaps. Calculation of maximum percent identity involves the production of an optimal alignment, taking into consideration gap penalties.
  • Suitable computer programs for carrying out sequence comparisons are widely available in the commercial and public sector. Examples include MatGat (Campanella et al., 2003,
  • sequence comparisons may be undertaken using the“needle” method of the EMBOSS Pairwise Alignment Algorithms, which determines an optimum alignment (including gaps) of two sequences when considered over their entire length and provides a percentage identity score.
  • Default parameters for amino acid sequence comparisons (“Protein Molecule” option) may be Gap Extend penalty: 0.5, Gap Open penalty: 10.0, Matrix: Blosum 62.
  • the sequence comparison may be performed over the full length of the reference sequence.
  • nucleic acid molecule which comprises a nucleotide sequence encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 4, and a polypeptide comprising an amino acid sequence of SEQ ID NO: 6.
  • nucleic acid molecule which comprises a nucleotide sequence encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 10, and a polypeptide comprising an amino acid sequence of SEQ ID NO: 12.
  • composition comprising a first nucleic acid which includes a nucleotide sequence encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 4, and a second nucleic acid which includes a nucleotide sequence encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 6.
  • composition comprising a first nucleic acid which includes a nucleotide sequence encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 10, and a second nucleic acid which includes a nucleotide sequence encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 12.
  • a combined preparation comprising: (i) a first nucleic acid which includes a nucleotide sequence encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 4; and (ii) a second nucleic acid which includes a nucleotide sequence encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 6.
  • a combined preparation comprising: (i) a first nucleic acid which includes a nucleotide sequence encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 10; and (ii) a second nucleic acid which includes a nucleotide sequence encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 12.
  • a composition comprising a first polypeptide comprising an amino acid sequence of SEQ ID NO: 4, and a second polypeptide comprising an amino acid sequence of SEQ ID NO: 6,
  • composition comprising a first polypeptide comprising an amino acid sequence of SEQ ID NO: 10, and a second polypeptide comprising an amino acid sequence of SEQ ID NO: 12.
  • fusion protein comprising a first polypeptide comprising an amino acid sequence of SEQ ID NO: 4, and a second polypeptide comprising an amino acid sequence of SEQ ID NO: 6.
  • fusion protein comprising a first polypeptide comprising an amino acid sequence of SEQ ID NO: 10, and a second polypeptide comprising an amino acid sequence of SEQ ID NO: 12.
  • a combined preparation comprising: (i) a first polypeptide comprising an amino acid sequence of SEQ ID NO: 4; and (ii) a second polypeptide comprising an amino acid sequence of SEQ ID NO: 6.
  • a combined preparation comprising: (i) a first polypeptide comprising an amino acid sequence of SEQ ID NO: 10; and (ii) a second polypeptide comprising an amino acid sequence of SEQ ID NO: 12.
  • combined preparation refers to a "kit of parts” in the sense that the combination components (i) and (ii) as defined above can be dosed independently or by use of different fixed combinations with distinguished amounts of the combination components (i) and (ii).
  • the components can be administered simultaneously or one after the other. If the components are administered one after the other, preferably the time interval between administration is chosen such that the therapeutic effect of the combined use of the components is greater than the effect which would be obtained by use of only any one of the combination components (i) and (ii).
  • the components of the combined preparation may be present in one combined unit dosage form, or as a first unit dosage form of component (i) and a separate, second unit dosage form of component (ii).
  • the ratio of the total amounts of the combination component (i) to the combination component (ii) to be administered in the combined preparation can be varied, for example in order to cope with the needs of a patient sub-population to be treated, or the needs of the single patient, which can be due, for example, to the particular disease, age, sex, or body weight of the patient.
  • there is at least one beneficial effect for example an enhancing of the effect of component (i), or component (ii), or a mutual enhancing of the effect of the combination components (i) and (ii), for example a more than additive effect, additional advantageous effects, fewer side effects, less toxicity, or a combined therapeutic effect compared with an effective dosage of one or both of the combination components (i) and (ii), and very preferably a synergism of the combination components (i) and (ii).
  • beneficial effect for example an enhancing of the effect of component (i), or component (ii), or a mutual enhancing of the effect of the combination components (i) and (ii), for example a more than additive effect, additional advantageous effects, fewer side effects, less toxicity, or a combined therapeutic effect compared with an effective dosage of one or both of the combination components (i) and (ii), and very preferably a synergism of the combination components (i) and (ii).
  • a combined preparation of the invention may be provided as a pharmaceutical combined preparation for administration to a mammal, preferably a human.
  • Component (i) may optionally be provided together with a pharmaceutically acceptable carrier, excipient, or diluent, and/or component (ii) may optionally be provided together with a pharmaceutically acceptable carrier, excipient, or diluent.
  • nucleic acid molecule encoding an amino acid sequence encoded by a nucleic acid of the invention.
  • nucleic acid molecule encoding a polypeptide of the invention.
  • a vector comprising a nucleic acid of the invention.
  • the vector further comprises a promoter operably linked to the nucleic acid.
  • the promoter is for expression of a polypeptide encoded by the nucleic acid in mammalian cells.
  • the promoter is for expression of a polypeptide encoded by the nucleic acid in yeast, bacterial, or insect cells.
  • the vector is a vaccine vector.
  • the vaccine vector is a viral vaccine vector, a bacterial vaccine vector, or a nucleic acid vector (for example an RNA vaccine vector, or a DNA vaccine vector).
  • a nucleic acid molecule of the invention may comprise a DNA or an RNA molecule.
  • the nucleic acid molecule comprises an RNA molecule
  • the molecule may comprise an RNA sequence that is at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical with, or identical with, any of SEQ ID NOs: 1 , 2, 4, 5, 7, 8, 10, 12, 14, 19, 21 , 23, 25, 27, 29, or 31 , in which each T nucleotide is replaced by IT, or the complement thereof.
  • the nucleic acid sequence of the nucleic acid of the invention will be an RNA sequence, so may comprise for example an RNA nucleic acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical with, or identical with, any of SEQ ID NOs: 1 , 2, 4, 5, 7, 8, 10, 12, 14, 19, 21 , 23, 25, 27, 29, or 31 in which each‘T’ nucleotide is replaced by 'Ll’, or the complement thereof.
  • Viral vaccine vectors use live viruses to carry nucleic acid (for example, DNA or RNA) into human or non-human animal cells.
  • the nucleic acid contained in the virus encodes one or more antigens that, once expressed in the infected human or non-human animal cells, elicit an immune response. Both humoral and cell-mediated immune responses can be induced by viral vaccine vectors.
  • Viral vaccine vectors combine many of the positive qualities of nucleic acid vaccines with those of live attenuated vaccines.
  • viral vaccine vectors carry nucleic acid into a host cell for production of antigenic proteins that can be tailored to stimulate a range of immune responses, including antibody, T helper cell (CD4 + T cell), and cytotoxic T lymphocyte (CTL, CD8 + T cell) mediated immunity.
  • Viral vaccine vectors unlike nucleic acid vaccines, also have the potential to actively invade host cells and replicate, much like a live attenuated vaccine, further activating the immune system like an adjuvant.
  • a viral vaccine vector therefore generally comprises a live attenuated virus that is genetically engineered to carry nucleic acid (for example, DNA or RNA) encoding protein antigens from an unrelated organism.
  • viral vaccine vectors are generally able to produce stronger immune responses than nucleic acid vaccines, for some diseases viral vectors are used in combination with other vaccine technologies in a strategy called heterologous prime-boost.
  • one vaccine is given as a priming step, followed by vaccination using an alternative vaccine as a booster.
  • the heterologous prime-boost strategy aims to provide a stronger overall immune response.
  • Viral vaccine vectors may be used as both prime and boost vaccines as part of this strategy. Viral vaccine vectors are reviewed by Lira et al., 2014 ( Vaccines 2014, 2, 624- 641 ) and Choi and Chang, 2013 (Clinical and Experimental Vaccine Research 2013;2:97- 105).
  • the viral vaccine vector is based on a viral delivery vector, such as a Poxvirus (for example, Modified Vaccinia Ankara (MVA), NYVAC, AVIPOX), herpesvirus (e.g. HSV, CMV, Adenovirus of any host species), Morbillivirus (e.g. measles), Alphavirus (e.g. SFV, Sendai), Flavivirus (e.g. Yellow Fever), or Rhabdovirus (e.g. VSV)-based viral delivery vector, a bacterial delivery vector (for example, Salmonella, E.coli), an RNA expression vector, or a DNA expression vector.
  • a viral delivery vector such as a Poxvirus (for example, Modified Vaccinia Ankara (MVA), NYVAC, AVIPOX), herpesvirus (e.g. HSV, CMV, Adenovirus of any host species), Morbillivirus (e.g. measles), Alphavirus (e.g. SFV, Send
  • an isolated cell comprising or transfected with a vector of the invention.
  • fusion protein comprising a polypeptide of the invention.
  • composition comprising a nucleic acid of the invention, and a pharmaceutically acceptable carrier, excipient, or diluent.
  • composition comprising a vector of the invention, and a pharmaceutically acceptable carrier, excipient, or diluent.
  • composition comprising a polypeptide of the invention, and a pharmaceutically acceptable carrier, excipient, or diluent.
  • composition of the invention further comprises an adjuvant for enhancing an immune response in a subject to the polypeptide, or to a polypeptide encoded by the nucleic acid, of the composition.
  • a method of inducing an immune response to a virus of the Filoviridae or Arenaviridae family in a subject which comprises administering to the subject a nucleic acid of the invention, a polypeptide of the invention, a vector of the invention, or a pharmaceutical composition of the invention.
  • a method of immunizing a subject against a virus of the Filoviridae or Arenaviridae family which comprises administering to the subject a nucleic acid of the invention, a polypeptide of the invention, a vector of the invention, or a pharmaceutical composition of the invention.
  • the pathogen is a virus.
  • the virus is a member of the Filoviridae, Arenaviridae, or Orthomyxoviridae family.
  • a method of inducing an immune response to a virus of the Filoviridae or Arenaviridae family in a subject which comprises administering to the subject a nucleic acid of the invention, a polypeptide of the invention, a vector of the invention, or a pharmaceutical composition of the invention.
  • a method of immunizing a subject against a pathogen which comprises administering to the subject a nucleic acid of the invention, a polypeptide of the invention, a vector of the invention, or a pharmaceutical composition of the invention.
  • the pathogen is a virus.
  • the virus is a member of the Filoviridae, Arenaviridae, or Orthomyxoviridae family.
  • a method of immunizing a subject against a virus of the Filoviridae family which comprises administering to the subject a nucleic acid of the invention, a polypeptide of the invention, a vector of the invention, or a pharmaceutical composition of the invention.
  • a method of inducing an immune response to a virus of the Filoviridae family in a subject which comprises administering to the subject a nucleic acid of the invention, a polypeptide of the invention, a vector of the invention, or a pharmaceutical composition of the invention.
  • nucleic acid, vector, or pharmaceutical composition of the invention comprises a nucleic acid comprising a sequence that is at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical with, or identical with, any of SEQ ID NOs:1 , 3, 5,
  • 7, 9, or 1 1 or comprises a nucleic acid encoding an amino acid sequence encoded by a nucleic acid comprising a sequence that is at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical with, or identical with, any of SEQ ID NOs: 1 , 3, 5, 7, 9, or 1 1.
  • polypeptide, vector, or pharmaceutical composition of the invention comprises a polypeptide comprising an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical with, or identical with, an amino acid sequence encoded by any of SEQ ID NOs: 1 , 3, 5, 7, 9, or 1 1 , or comprises a polypeptide comprising an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical with, or identical with, any of SEQ ID NOs: 2, 4, 6, 8, 10, or 12.
  • a method of immunizing a subject against a virus of the Arenaviridae family which comprises administering to the subject a nucleic acid of the invention, a polypeptide of the invention, a vector of the invention, or a pharmaceutical composition of the invention.
  • a method of inducing an immune response to a virus of the Arenaviridae family in a subject which comprises administering to the subject a nucleic acid of the invention, a polypeptide of the invention, a vector of the invention, or a pharmaceutical composition of the invention.
  • nucleic acid, vector, or pharmaceutical composition of the invention comprises a nucleic acid comprising a sequence that is at least 75%, 80%, 85%, 90%,
  • polypeptide, vector, or pharmaceutical composition of the invention comprises a polypeptide comprising an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical with, or identical with, an amino acid sequence encoded by any of SEQ ID NOs: 14, 16, 18, 20, 22, 24, or 26, or comprises a polypeptide comprising an amino acid sequence that is at least 95%, 96%, 97%, 98%, or 99% identical with, or identical with, any of SEQ ID NOs: 13, 15, 17, 19, 21 , 23, or 25.
  • Methods of administration include, but are not limited to, intradermal, intramuscular, intraperitoneal, parenteral, intravenous, subcutaneous, vaginal, rectal, intranasal, inhalation or oral.
  • Parenteral administration such as subcutaneous, intravenous or intramuscular administration, is generally achieved by injection.
  • Injectables can be prepared in conventional forms, either as liquid solutions or suspensions, solid forms suitable for solution or suspension in liquid prior to injection, or as emulsions.
  • Injection solutions and suspensions can be prepared from sterile powders, granules, and tablets of the kind previously described.
  • Administration can be systemic or local.
  • Compositions may be administered in any suitable manner, such as with pharmaceutically acceptable carriers.
  • compositions are determined in part by the particular composition being administered, as well as by the particular method used to administer the composition.
  • Preparations for parenteral administration include sterile aqueous or nonaqueous solutions, suspensions, and emulsions.
  • non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate.
  • Aqueous carriers include water, alcoholic/aqueous solutions, emulsions or suspensions, including saline and buffered media.
  • Parenteral vehicles include sodium chloride solution, Ringer’s dextrose, dextrose and sodium chloride, lactated Ringer’s, or fixed oils.
  • Intravenous vehicles include fluid and nutrient replenishers, electrolyte replenishers (such as those based on Ringer's dextrose), and the like. Preservatives and other additives may also be present such as, for example, antimicrobials, anti-oxidants, chelating agents, and inert gases and the like.
  • compositions may potentially be administered as a pharmaceutically acceptable acid- or base-addition salt, formed by reaction with inorganic acids such as hydrochloric acid, hydrobromic acid, perchloric acid, nitric acid, thiocyanic acid, sulfuric acid, and phosphoric acid, and organic acids such as formic acid, acetic acid, propionic acid, glycolic acid, lactic acid, pyruvic acid, oxalic acid, malonic acid, succinic acid, maleic acid, and fumaric acid, or by reaction with an inorganic base such as sodium hydroxide, ammonium hydroxide, potassium hydroxide, and organic bases such as mono-, di-, trialkyl and aryl amines and substituted ethanolamines.
  • inorganic acids such as hydrochloric acid, hydrobromic acid, perchloric acid, nitric acid, thiocyanic acid, sulfuric acid, and phosphoric acid
  • organic acids such as formic acid, acetic acid, propionic acid
  • Administration can be accomplished by single or multiple doses.
  • the dose administered to a subject in the context of the present disclosure should be sufficient to induce a beneficial therapeutic response in a subject over time, or to inhibit or prevent infection.
  • the dose required will vary from subject to subject depending on the species, age, weight and general condition of the subject, the severity of the infection being treated, the particular composition being used and its mode of administration. An appropriate dose can be determined by one of ordinary skill in the art using only routine experimentation.
  • Pharmaceutically acceptable carriers include, but are not limited to, saline, buffered saline, dextrose, water, glycerol, ethanol, and combinations thereof.
  • the carrier and composition can be sterile, and the formulation suits the mode of administration.
  • the composition can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents.
  • the composition can be a liquid solution, suspension, emulsion, tablet, pill, capsule, sustained release formulation, or powder.
  • the composition can be formulated as a suppository, with traditional binders and carriers such as triglycerides.
  • Oral formulations can include standard carriers such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, and magnesium carbonate. Any of the common pharmaceutical carriers, such as sterile saline solution or sesame oil, can be used.
  • the medium can also contain conventional pharmaceutical adjunct materials such as, for example, pharmaceutically acceptable salts to adjust the osmotic pressure, buffers, preservatives and the like.
  • Other media that can be used with the compositions and methods provided herein are normal saline and sesame oil.
  • compositions comprise a pharmaceutically acceptable carrier and/or an adjuvant.
  • the adjuvant can be alum, Freund’s complete adjuvant, a biological adjuvant or immunostimulatory oligonucleotides (such as CpG oligonucleotides).
  • parenteral formulations usually comprise injectable fluids that include pharmaceutically and physiologically acceptable fluids such as water, physiological saline, balanced salt solutions, aqueous dextrose, glycerol or the like as a vehicle.
  • pharmaceutically and physiologically acceptable fluids such as water, physiological saline, balanced salt solutions, aqueous dextrose, glycerol or the like as a vehicle.
  • physiologically acceptable fluids such as water, physiological saline, balanced salt solutions, aqueous dextrose, glycerol or the like
  • solid compositions for example, powder, pill, tablet, or capsule forms
  • conventional non-toxic solid carriers can include, for example, pharmaceutical grades of mannitol, lactose, starch, or magnesium stearate.
  • compositions to be administered can contain minor amounts of non-toxic auxiliary substances, such as wetting or emulsifying agents, preservatives, and pH buffering agents and the like, for example sodium acetate or sorbitan monolaurate.
  • non-toxic auxiliary substances such as wetting or emulsifying agents, preservatives, and pH buffering agents and the like, for example sodium acetate or sorbitan monolaurate.
  • composition of the invention is administered intramuscularly.
  • composition is administered intramuscularly, intradermaly, subcutaneously by needle or by gene gun, or electroporation.
  • a polypeptide of the invention may include one or more conservative amino acid substitutions.
  • Conservative amino acid substitutions are those substitutions that, when made, least interfere with the properties of the original protein, that is, the structure and especially the function of the protein is conserved and not significantly changed by such substitutions. Examples of conservative substitutions are shown below:
  • Conservative substitutions generally maintain (a) the structure of the polypeptide backbone in the area of the substitution, for example, as a sheet or helical conformation, (b) the charge or hydrophobicity of the molecule at the target site, or (c) the bulk of the side chain.
  • substitutions which in general are expected to produce the greatest changes in protein properties will be non-conservative, for instance changes in which (a) a hydrophilic residue, for example, seryl or threonyl, is substituted for (or by) a hydrophobic residue, for example, leucyl, isoleucyl, phenylalanyl, valyl or alanyl; (b) a cysteine or proline is substituted for (or by) any other residue; (c) a residue having an electropositive side chain, for example, lysyl, arginyl, or histidyl, is substituted for (or by) an electronegative residue, for example, glutamyl or aspartyl; or (d) a residue having a bulky side chain, for example, phenylalanine, is substituted for (or by) one not having a side chain, for example, glycine.
  • the embodiments described herein induce broad neutralisation profiles against the Ebolavirus genus
  • Figure 1 shows a phylogenetic tree comparing ebolaviruses and Marburg viruses. Numbers indicate percent confidence of branches;
  • FIG. 2 shows challenge study results for an Ebola challenge model.
  • Ebola challenge model was lethal for non-vaccinated guinea pigs (Group 1 , lower line) whereas all vaccinated guinea pigs (Group 2, upper line) were protected (left) and continued to gain weight (right); and
  • Figure 3 shows the results of a neutralisation assay illustrating the strength of neutralising antibody responses to expressed target antigens, representative of all Ebola virus species and Marburg viruses.
  • Strength of neutralisation is indicated by the heat-map where red (darkest shading when viewed in grayscale) is very strong neutralisation, decreasing through orange to yellow (progressively lighter shading when viewed in grayscale) and no neutralising/equal to negative control values are white.
  • T2-4 and T2-6 are nucleic acid vaccines Ebola polypeptide, combined with T2-1 1 a Marburg candidate, at pre-clinical stage testing with serum samples taken from immunised guinea pigs.
  • Example 1
  • Ebola polypeptides able to induce a broadly neutralizing antibody response
  • Table 1 below shows flow cytometric assay results illustrating the strength of antibody binding to target antigens, representative of all Ebola virus species (subtypes) and Marburg viruses. Strength of binding is indicated by the heat-map where red (the darkest shading when viewed in grayscale) is very strong binding, decreasing through orange to yellow (progressively lighter shading when viewed in grayscale) and no binding/equal to negative control values are white. Serum samples 1 -22 were taken from individuals immunised with other Ebola virus vaccine candidates. T2-4 and T2-6 are nucleic acid vaccines encoding lead candidate optimized antigenic Ebola polypeptide, combined with T2-1 1 a Marburg candidate, at pre-dinical stage testing with serum samples taken from immunised guinea pigs.
  • Tri-LEMvac Trivalent vaccine
  • the Modified Vaccinia Ankara (MVA) vaccine platform is a nonreplicating strain (i.e. non-replicating in human cells), third generation smallpox vaccine and one of the most advanced recombinant poxviral vaccine vectors in human clinical trials (Cottingham & Carroll, Vaccine, 2013, 31 (39):4247-51 ).
  • MVA is a robust vector system capable of co-expressing up to four transgenes facilitating potent promoters and stable insertion sites (Orubu et ai, Pone, 2012,7(6)e0040167).
  • MVA was chosen because: 1 ) its significant capacity to stably express multiple independent ORFs via compatible expression cassettes with strong and timely regulated promotors for trivalent LEM vaccination in one cost effective vaccine lot; 2) its ability to induce robust B and T-cell immune responses in animals and humans especially when primed or boosted with DNA or RNA vectors; and 3) vaccine lots can be thermally stabilised for storage and transport in developing countries in the absence of cold chain (Frey et ai, Vaccine, 2015, 33(39):5225-34).
  • Proof of principle for the Trivalent vaccine candidate has been demonstrated by: i) cassette validation for independent L, E and M GPC expression and epitope presentation; and ii) preclinical efficacy by Filovirus challenge.
  • the challenge study results are shown in Figure 2.
  • the Ebola challenge model was lethal for non-vaccinated guinea pigs (Group 1 , lower line) whereas all vaccinated guinea pigs (Group 2, upper line) were protected (left) and
  • Figure 3 shows the results of a neutralisation assay illustrating the strength of neutralising antibody responses to surface-expressed target antigens, representative of all Ebola virus species and Marburg viruses.
  • Strength of neutralisation is indicated by the heat-map where red (darkest shading when viewed in grayscale) is very strong neutralisation, decreasing through orange to yellow (progressively lighter shading when viewed in grayscale) and no neutralising/equal to negative control values are white.
  • T2-4 and T2-6 are nucleic acid vaccines each encoding lead candidate optimized antigenic Ebola polypeptide, combined with T2-1 1 a Marburg candidate, at pre-clinical stage testing with serum samples taken from immunised guinea pigs.
  • Lassa fever is a hemorrhagic disease caused by an Old World (OW) arenavirus known as Lassa virus (LASV).
  • OW Old World
  • LASV Lassa virus
  • the virus was first isolated in Nigeria in 1969 and is currently endemic in West Africa. Due to the high morbidity and mortality associated with Lassa hemorrhagic fever, LASV is classified as a category A pathogen.
  • Lassa virus is an enveloped ambisense RNA virus with a bisegmented genome. Viral particles are covered in mature glycoprotein (GP) trimeric spikes, which mediate viral entry. Like other class 1 viral fusion proteins, the envelope glycoprotein precursor (GPC) is translated as a single polypeptide and is proteolytically cleaved into three subunits.
  • GP mature glycoprotein
  • GPC endoplasmic reticulum
  • SKI-1/S1 P cellular proprotein convertase subtilisin kexin isozyme- 1 /site- 1 protease
  • SSP stable-signal peptide
  • GPC noncovalent stable-signal peptide
  • GPC noncovalent stable-signal peptide
  • SSP interacts with the cytoplasmic domain of GP2 and is involved in pH sensing.
  • GP1 is responsible for binding to cellular receptors, while GP2 mediates membrane fusion during viral entry.
  • construct 1 was identified. Modifications were then introduced independently into the parental sequence (construct 1 ) to provide: (A) SOSEP (construct 2); and (B) FLEP
  • construct 4 construct 4
  • NtoK glycan knock-out
  • A Two cystein residues were introduced at positions 207 and 360 to allow formation of a disulfide bridge (SOS) between the exterior and the transmembrane domains of GP. To facilitate complete cleavage of these two domains, the furin cleavage site was modified from RRLL to RRRR at position 256-259. Mutation of glutamate to proline at position 329 (EP) prevents structural rearrangements making the protein less flexible.
  • SOS disulfide bridge
  • Variants of both designs were generated that additionally contain an asparagine to lysine mutation at position 272 or 278, for SOSEP-NtoK or FLEP-NtoK, respectively, to inactivate a glycosylation motif. Glycans at this position might block access of some neutralizing antibodies, such as 37.7H.
  • Amino acid sequence (SEQ ID NO:21 ):
  • This example describes optimized Lassa virus nucleoprotein sequence according to an embodiment of the invention.
  • This example describes optimized Lassa virus nucleoprotein sequence according to an embodiment of the invention.

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Abstract

L'invention concerne des molécules d'acide nucléique, des polypeptides, des vecteurs, des cellules, des protéines de fusion, des compositions pharmaceutiques et leur utilisation en tant que vaccins contre des pathogènes, en particulier contre des pathogènes émergents ou ré-émergents (en particulier des virus à ARN).
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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7731975B2 (en) * 2001-01-31 2010-06-08 The United States Of America As Represented By The Secretary Of The Army Chimeric filovirus glycoprotein
WO2016097065A1 (fr) * 2014-12-16 2016-06-23 Curevac Ag Vaccins contre le virus ebola et le virus marburg
WO2017172622A1 (fr) * 2016-03-28 2017-10-05 Integrated Biotherapeutics, Inc. Compositions de vaccin contre le pan-filovirus et leurs procédés de fabrication
WO2018115525A1 (fr) * 2016-12-23 2018-06-28 Curevac Ag Vaccin contre le virus de lassa

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Publication number Priority date Publication date Assignee Title
US7731975B2 (en) * 2001-01-31 2010-06-08 The United States Of America As Represented By The Secretary Of The Army Chimeric filovirus glycoprotein
WO2016097065A1 (fr) * 2014-12-16 2016-06-23 Curevac Ag Vaccins contre le virus ebola et le virus marburg
WO2017172622A1 (fr) * 2016-03-28 2017-10-05 Integrated Biotherapeutics, Inc. Compositions de vaccin contre le pan-filovirus et leurs procédés de fabrication
WO2018115525A1 (fr) * 2016-12-23 2018-06-28 Curevac Ag Vaccin contre le virus de lassa

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