Influenza h5 vaccines

Abstract

The present invention is based on the surprising finding that H5 protein of clade 1 H5N1 induces, in particular by a single-shot vaccination, a cross-clade protective immune response to influenza viruses with H5N1 HA. In one aspect, the invention is thus directed to H5 protein of clade 1 H5N1 virus for use in a method of treating or preventing infections with H5N1 virus of a different clade, namely of a clade different from clade 1 or from any clade with the exception of clade 1, respectively.

1 . An H5 protein of a clade 1 H5N1 virus for use in a method of treating or preventing infections with a H5N1 virus of a different clade, wherein said H5 protein comprises a polypeptide sequence having at least 98% sequence identity with the polypeptide sequence of SEQ ID NO: 1. 
     
     
         2 . The H5 protein according to  claim 1 , wherein said H5 protein comprises a polypeptide sequence having at least 98.1%, preferably at least 98.2%, more preferably at least 98.3%, and most preferably at least 98.4% sequence identity with the polypeptide sequence of SEQ ID NO: 1. 
     
     
         3 . The H5 protein according to  claim 1 , wherein said H5 protein has the amino acid 223N and the modification 328K+, wherein numbering of the amino acid positions of the H5 protein refers to the amino acid position as exemplarily given in SEQ ID NO:2, and wherein the modification 328K+ means that at amino acid position 328 of H5 protein a second Lysine (K+) is inserted. 
     
     
         4 . The H5 protein according to  claim 1 , wherein such H5 protein has the amino acid 94N. 
     
     
         5 . The H5 protein according to  claim 1 , wherein such H5 protein has the amino acid 120N. 
     
     
         6 . The H5 protein according to  claim 1 , wherein such H5 protein has the amino acid 155N. 
     
     
         7 . The H5 protein according to  claim 1 , wherein such H5 protein has one or more of the following amino acid clusters selected from the group consisting of:
 a. aa 93-95: GNF   b. aa 123-125: SDH   c. aa 128-130: SSG   d. aa 138-140: GSS   e. aa 226-228: MDF   f. aa 270-272: EVE   g. aa 309-311: NKL.   
     
     
         8 . The H5 protein according to  claim 1 , wherein such H5 protein comprises a peptide comprising:
 i. the amino acid sequences of SEQ ID NO:5; SEQ ID NO:6 or SEQ ID NO:7; or   ii. any peptide that has at least 85% sequence homology to the polypeptide of i) and that comprises hemagglutinin inhibition in a standard hemagglutinin inhibition assay; or   iii. any part of the polypeptides of i) or ii) comprising at least 8 contiguous amino acids of any of such peptides of i) or ii) and wherein any of such peptide comprises hemagglutinin inhibition in a standard hemagglutinin inhibition assay; or   iv. any peptide of i), ii) or iii) having one of the amino acids 36T, 36K, 83A, 83T, 83D, 86A, 86V, 120S, 155S, 156A, 156T, 189R, 189K, 212K, 212R, 212E, 263A or 263T; or   v. any peptide of i), ii), iii) or iv) having one or more of the following amino acid clusters selected from the group consisting of:
 a. aa 93-95: GNF 
 b. aa 123-125: SDH 
 c. aa 128-130: SSG 
 d. aa 138-140: GSS 
 e. aa 226-228: MDF 
 f. aa 270-272: EVE 
 g. aa309-311: NKL. 
   
     
     
         9 . The H5 protein according to  claim 1 , wherein such H5 protein comprises the amino acid sequence of SEQ ID NO:5. 
     
     
         10 . The H5 protein according to  claim 1 , wherein such H5 protein is recombinantly expressed and/or produced by a baculovirus expression system, preferably in cultured insect cells. 
     
     
         11 . The H5 protein according to  claim 1 , wherein said H5N1 virus of a different clade is selected from the group consisting of clade 0 H5N1 virus, clade 2 H5N1 virus, clade 3 H5N1 virus, clade 4 H5N1 virus, clade 5 H5N1 virus, clade 6 H5N1 virus, clade 7 H5N1 virus, clade 8 H5N1 virus and clade 9 H5N1 virus. 
     
     
         12 . The H5 protein according to  claim 1 , wherein said H5N1 virus of a different clade is clade 2.2 H5N1 virus or a clade 2.3 H5N1 virus. 
     
     
         13 . The H5 protein according to  claim 1 , wherein said H5N1 virus of a different clade is a clade 2.2.1 H5N1 virus or a clade 2.3.2 H5N1 virus. 
     
     
         14 . The H5 protein according to  claim 1 , wherein said H5N1 virus of a different clade is a H5N1 virus of North African or of Vietnamese origin. 
     
     
         15 . The H5 protein according to  claim 14 , wherein said H5N1 virus of North African origin is a H5N1 virus comprising a second H5 protein of influenza virus, wherein said second H5 protein encodes an amino acid sequence including at least one member of a group consisting of:
 (a) the amino acids 113D, 126H, 145(−), 156R, 160F, 167T, and 181N, wherein the modification 145(−) means that amino acid position 145 of H5 is deleted, or   (b) the amino acids 87P, 145L, 172T, 201E, 206I, 208K, 254T, 341G and 421K, or   (c) the amino acids 145L, 172T, and 254V,   and wherein the numbering of the amino acid positions of said second H5 protein refers to the amino acid position as exemplarily given in SEQ ID NO:8;   or wherein said second H5 protein consists of an amino acid sequence which is at least 95%, preferably at least 96%, more preferably at least 97%, still more preferably at least 98%, yet more preferably at least 99%, or in particular preferred 100% homolog with any one of the sequences as set forth in SEQ ID NOs: 9 to 46.   
     
     
         16 . The H5 protein according to  claim 1 , wherein said H5N1 virus of a different clade comprising a second H5 protein encodes an amino acid sequence including at least one member of the group consisting of:
 (a) the amino acids 87L, 113D, 126H, 145(−), 156R, 160F, 167T, and 181N, or   (b) the amino acids 87P, 113N, 126R, 145L, 160Y, 172T, 181H, 201E, 206I, 208K, 254T, 341G and 421K, or   (c) the amino acids 87L, 113N, 126R, 145L, 156G, 160Y, 172T, 181H, and 254V,   and/or wherein such second H5 protein comprises a peptide comprising:   i. any one of the amino acid sequences of SEQ ID NOs: 9 to 46;   ii. any peptide that has at least 85%, preferably at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, even more preferably at least 99%, most preferably 100% sequence homology to the polypeptide of i) and that comprises hemagglutinin inhibition in a standard hemagglutinin inhibition assay; or   iii. any part of the polypeptides of i) or ii) comprising at least 334 contiguous amino acids of any of such peptides of i) or ii) and wherein any of such peptide comprises hemagglutinin inhibition in a standard hemagglutinin inhibition assay,   and/or wherein such second H5 protein comprises a contiguous amino acid sequence which has at least 95%, even more preferably at least 96%, even more preferably at least 97%, even more preferably at least 98%, even more preferably at least 99%, most preferably 100% sequence identity with any one of the sequences as set forth in SEQ ID NOs: 9 to 46.   
     
     
         17 . The H5 protein according to  claim 1 , wherein said H5N1 virus of a different clade comprising a second H5 protein consists of an amino acid sequence which is at least 95%, preferably at least 96%, more preferably at least 97%, still more preferably at least 98%, yet more preferably at least 99%, or in particular preferred 100% homolog with any one of the sequences as set forth in SEQ ID NOs: 15 or 20, and wherein such second H5 protein consisting of the amino acid sequence set forth in SEQ ID NO:20 is more preferred. 
     
     
         18 . The H5 protein according to  claim 1 , for use in a method of treating or preventing viral infection, wherein said viral infection includes a virus from at least one member of a group selected from:
 (A) Subclade A H5N1 virus of North African origin, namely an infection with a H5N1 virus comprising a second H5 protein encoded by the amino acid sequence according to at least one of the sequences as set forth in SEQ ID NOs: 9 to 19, or 42 or 43,   or   (B) with Subclade B H5N1 virus of North African origin, namely an infection with a H5N1 virus comprising a H5 protein encoded by the amino acid sequence according to at least one of the sequences as set forth in SEQ ID NOs: 20 to 41, or 44 to 46.   
     
     
         19 . A combination of an H5 protein of a clade 1 H5N1 virus for use in a method of treating or preventing infections with a H5N1 virus of a different clade, wherein said H5 protein comprises a polypeptide sequence having at least 98% sequence identity with the polypeptide sequence of SEQ ID NO: 1 and an inactivated Newcastle disease virus. 
     
     
         20 . The combination of  claim 19 , wherein the inactivated Newcastle disease virus is an inactivated whole Newcastle disease virion. 
     
     
         21 . The combination of  claim 19 , wherein the inactivated Newcastle disease virus is an inactivated Newcastle disease virus obtained by inactivation of a Newcastle disease virus comprising a RNA polynucleotide having at least 70%, preferably at least 80%, more preferably at least 90%, still more preferably at least 95% or in particular 100% sequence identity with a RNA copy of the polynucleotide set forth in SEQ ID NO: 51, which has been inactivated. 
     
     
         22 . The combination of  claim 19 , wherein the Newcastle disease virus is a Newcastle disease LaSota strain virus. 
     
     
         23 . The combination of  claim 19 , wherein the Newcastle Disease Virus is inactivated with a reagent selected from the group consisting of Formaldehyde, BEI, Beta-Propio-Lactone (BPL), and combinations thereof. 
     
     
         24 . A vaccine for use in a method of treating or preventing infections with a H5N1 virus, comprising:
 a. the H5 protein of a different clade, wherein said H5 protein comprises a polypeptide sequence having at least 98% sequence identity with the polypeptide sequence of SEQ ID NO:1 of, and   b. a pharmaceutical acceptable carrier and/or excipient.   
     
     
         25 . The vaccine according to  claim 24 , wherein the excipient is one or more adjuvants. 
     
     
         26 . The vaccine according to  claim 25 , wherein the adjuvant is an Emulsigen-based adjuvant. 
     
     
         27 . The vaccine according to  claim 24 , wherein the vaccine comprises one or more further antigens. 
     
     
         28 . The vaccine according to  claim 27 , wherein the one or more further antigen is an antigen of a poultry pathogen. 
     
     
         29 . The vaccine according to  claim 28 , wherein the one or more further antigen is H5, H7, or H9 of influenza virus. 
     
     
         30 . The vaccine according to  claim 29 , wherein the H5 of influenza virus is H5 protein of a H5N1 virus of a clade different than clade 1. 
     
     
         31 . Use of the H5 protein wherein said H5 protein comprises a polypeptide sequence having at least 98% sequence identity with the polypeptide sequence of SEQ ID NO:1, for the preparation of a pharmaceutical composition, preferably of a single-shot vaccine or a one dose vaccine, for the prophylaxis or treatment of infections caused by H5N1 virus of a clade other than clade 1. 
     
     
         32 . A method for the treatment or prophylaxis of influenza virus infections caused by H5N1 virus of a clade other than clade 1, wherein the method comprises administration of a therapeutically effective amount of the H5 protein, wherein said H5 protein comprises a polypeptide sequence having at least 98% sequence identity with the polypeptide sequence of SEQ ID NO: 1, to a subject in need of such a treatment. 
     
     
         33 . A method for the treatment or prophylaxis of influenza virus infections caused by H5N1 virus of a clade other than clade 1, wherein the method comprises administration of a therapeutically effective amount of a vaccine comprising:
 a. the H5 protein of a different clade, wherein said H5 protein comprises a polypeptide sequence having at least 98% sequence identity with the polypeptide sequence of SEQ ID NO: 1 of, and   b. a pharmaceutical acceptable carrier and/or excipient   to a subject in need of such a treatment.   
     
     
         34 . The method of  claim 32  wherein said administration is a single-shot administration or a one dose administration. 
     
     
         35 . A kit of parts, comprising:
 a. the H5 protein according to  claim 1 ; and   b. a package leaflet indicating the use of such H5 protein, combination or vaccine of a) for the treatment or prophylaxis of infections caused by H5N1 virus of a clade other than clade 1.   
     
     
         36 . The kit according to  claims 35 , wherein such kit comprises at least one or more further antigens of poultry or mammalian pathogen. 
     
     
         37 . (canceled) 
     
     
         38 . A method of reducing the incidence of or severity of influenza infection comprising the step of administering a composition selected from the group consisting of:
 a. an H5 protein of a clade 1 H5N1 virus, wherein said H5 protein comprises a polypeptide sequence having at least 98% sequence identity with the polypeptide sequence of SEQ ID NO: 1, and   b. a combination of the H5 protein of group a) and an inactivated Newcastle disease virus.   
     
     
         39 . (canceled) 
     
     
         40 . Use of the H5 protein wherein said H5 protein comprises a polypeptide sequence having at least 98% sequence identity with the polypeptide sequence of SEQ ID NO:1, for the preparation of a medicament for reducing viral shedding in a subject infected with or at risk of a viral infection with H5N1 virus of a clade other than clade 1. 
     
     
         41 . A vaccine for use in a method of treating or preventing infections with a H5N1 virus, comprising:
 a. the combination of the H5 protein of a different clade and an inactivated Newcastle disease virus, and   b. a pharmaceutical acceptable carrier and/or excipient.   
     
     
         42 . The vaccine according to  claim 41 , wherein the excipient is one or more adjuvants. 
     
     
         43 . The vaccine according to  claim 41 , wherein the adjuvant is an Emulsigen-based adjuvant. 
     
     
         44 . The vaccine according to  claim 41 , wherein the vaccine comprises one or more further antigens. 
     
     
         45 . The vaccine according to  claim 44 , wherein the one or more further antigen is an antigen of a poultry pathogen. 
     
     
         46 . The vaccine according to  claim 45 , wherein the one or more further antigen is H5, H7, or H9 of influenza virus. 
     
     
         47 . The vaccine according to  claim 46 , wherein the H5 of influenza virus is H5 protein of a H5N1 virus of a clade different than clade 1. 
     
     
         48 . Use of the combination of the H5 protein wherein said H5 protein comprises a polypeptide sequence having at least 98% sequence identity with the polypeptide sequence of SEQ ID NO:1 and an inactivated Newcastle disease virus, for the preparation of a pharmaceutical composition, preferably of a single-shot vaccine or a one dose vaccine, for the prophylaxis or treatment of infections caused by H5N1 virus of a clade other than clade 1. 
     
     
         49 . A method for the treatment or prophylaxis of influenza virus infections caused by H5N1 virus of a clade other than clade 1, wherein the method comprises administration of a therapeutically effective amount of the combination of the H5 protein wherein said H5 protein comprises a polypeptide sequence having at least 98% sequence identity with the polypeptide sequence of SEQ ID NO:1 and an inactivated Newcastle disease virus combination to a subject in need of such a treatment. 
     
     
         50 . The method of  claim 41 , wherein said administration is a single-shot administration or a one dose administration. 
     
     
         51 . A method of reducing the incidence of or severity of influenza infection comprising the step of administering a composition selected from the group consisting of:
 a. an H5 protein of clade 1 H5N1 virus, wherein said H5 protein comprises or consists of a polypeptide sequence having at least 98% sequence identity with the polypeptide sequence of SEQ ID NO: 1, and   b. a combination of the H5 protein of group a) and an inactivated Newcastle disease virus.   
     
     
         52 . The H5 protein of  claim 1 , for use in a method for reducing viral shedding in a subject, wherein said H5 protein is to be administered to a subject infected with or at risk of a viral infection with H5N1 virus of a clade other than clade 1. 
     
     
         53 . The method of  claim 52 , further comprising use of an inactivated Newcastle disease virus in combination with the H5 protein. 
     
     
         54 . Use of a H5 protein wherein said H5 protein comprises a polypeptide sequence having at least 98% sequence identity with the polypeptide sequence of SEQ ID NO:1, for the preparation of a medicament for reducing viral shedding in a subject infected with or at risk of a viral infection with H5N1 virus of a clade other than clade 1. 
     
     
         55 . The medicament of  claim 54 , further comprising use and an inactivated Newcastle disease virus in combination with the H5 protein, wherein the combination reduces viral shedding in a subject infected with or at risk of a viral infection with H5N1 virus of a clade other than clade 1.