PEPTIDE VACCINE FOR INFLUENZA VIRUS
First Claim
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1. A method for evaluating the potential of a chemical entity to bind to a peptide epitope derived from the divalent sialoside binding site of hemagglutinin protein of influenza virus comprising the steps of:
- (i) contacting said chemical entity with said peptide under conditions that allow said chemical entity to bind said peptide; and
(ii) detecting the presence of a complex of said chemical entity and said peptide;
wherein said peptide epitope is peptide 1 corresponding to cysteine 97 region, and/or peptide 2 corresponding to cysteine 139 region and/or peptide 3 corresponding to the region of amino acids 220-226 as defined by the amino acid sequence of X31-hemaglutinin andwhereinsaid peptide epitope comprisesa) a conformational peptide epitope, comprising at least one cysteine residue or cysteine analogous amino acid residue conjugated from the side chain and the peptide epitope comprises less than 100 amino acid residues, preferably less than 30 amino acid residues present in a natural influenza virus peptide orb) the conformational peptide epitope is a short peptide epitope comprising 3 to 12 amino acid residues, preferably comprising less than 12 amino acid residues, more preferably less than 11 amino acid residue.
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Abstract
The invention relates to the method for evaluating the potential of a chemical entity, such as an antibody, to bind to a peptide epitope derived from the divalent sialoside binding site of hemagglutinin protein of influenza virus. The invention also provides peptide epitopes 5 for use in the prevention and/or treatment of influenza or for the development of such treatment or vaccine against influenza.
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Citations
97 Claims
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1. A method for evaluating the potential of a chemical entity to bind to a peptide epitope derived from the divalent sialoside binding site of hemagglutinin protein of influenza virus comprising the steps of:
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(i) contacting said chemical entity with said peptide under conditions that allow said chemical entity to bind said peptide; and (ii) detecting the presence of a complex of said chemical entity and said peptide; wherein said peptide epitope is peptide 1 corresponding to cysteine 97 region, and/or peptide 2 corresponding to cysteine 139 region and/or peptide 3 corresponding to the region of amino acids 220-226 as defined by the amino acid sequence of X31-hemaglutinin and wherein said peptide epitope comprises a) a conformational peptide epitope, comprising at least one cysteine residue or cysteine analogous amino acid residue conjugated from the side chain and the peptide epitope comprises less than 100 amino acid residues, preferably less than 30 amino acid residues present in a natural influenza virus peptide or b) the conformational peptide epitope is a short peptide epitope comprising 3 to 12 amino acid residues, preferably comprising less than 12 amino acid residues, more preferably less than 11 amino acid residue. - View Dependent Claims (6, 8, 10, 11, 14, 21, 26, 28, 33, 38, 40, 42, 43, 57, 69, 70, 78, 88, 97)
and wherein said large binding site is optionally further defined by at least one of the structure coordinates of influenza hemagglutinin semi- or nonconserved amino acids Gly134, Asn137, Ala138, Thr155, Glu190 and Leu226 of Region A;
Phe94, Asn96, Asn137, Ala138, Lys140 and Arg207 of Region B;
Ser91, Ala 93, Tyr105 and Arg208 of Region C.
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38. The method according to the claim 1, for identifying a modulator of binding between the large binding site of influenza hemagglutinin and its ligand divalent sialoside, comprising steps of:
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(a) contacting the large binding site of influenza hemagglutinin and its ligand in the presence and in the absence of a putative modulator compound; (b) detecting binding between the large binding site of influenza hemagglutinin and its ligand in the presence and absence of the putative modulator; and (c) identifying a modulator compound in view of decreased or increased binding between the large binding site of influenza hemagglutinin and its ligand in the presence of the putative modulator, as compared to binding in the absence of the putative modulator, wherein the modulator binds to peptide epitope according to claim 1.
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40. The method according to the claim 1, for selecting peptide epitopes for immunization and developing peptide vaccines against influenza comprising at least one di- to decapeptide epitope of the large binding site described in Table 1, wherein the method involves analysis according to the claim 1 for antibody as a chemical entity blocking the large binding site.
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42. The method according to claim 1, using peptide 1, peptide 2 or peptide 3 selected from the group consisting of
K1V2R3, W1V2R3, K1V2N3, T1P2N3P4E5N6G7T8, S1K2A3Y4S5N6, K1A2N3P4A5N6D7L8, V1T2K3G4V5S6A7S8, G1T2S3SAA5, E1A2S3S4G5V6S7S8A9, and said peptide corresponding to influenza virus A hemagglutinin. -
43. The antigenic compound according to the claim 10 comprising a peptide selected from the group consisting of
K1V2R3, W1V2R3, K1V2N3, T1P2N3P4E5N6G7T8, S1K2A3Y4S5N6, K1A2N3P4A5N6D7L8, V1T2K3G4V5S6A7S8, G1T2S3S4A5, E1A2S3S4G5V6S7S8A9, and said peptide corresponds to influenza virus A hemagglutinin. -
57. The antigenic compound according to claim 1, wherein said antigenic compound comprises at least two peptides as defined in claim 1.
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69. An isolated nucleotide encoding an antigenic compound as described in claim 1, the substance optionally being a primer.
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70. The nucleotide according to the claim 69 for use in the method for detecting nucleic acid encoding antigenic compound according to claim 43 in a sample comprising:
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amplifying DNA reverse transcribed from RNA obtained from the sample using one or more primers each comprising any one of the sequences as listed in Table 1 or sequences in FIGS. 17-19 ;and detecting a product of amplification, wherein the presence of the product of amplification indicates the presence of an influenza virus hemagglutinin in the sample.
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78. The nucleotide according to the claim 69, for a use of detecting nucleic acid encoding antigenic compound according to claim 1 in a sample comprising:
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contacting the sample with a primer immobilized on a support, said primer comprising a sequence listed in Table 1 or sequences in FIGS. 17-19 , under conditions suitable for hybridizing the primer and the sample; anddetecting hybridization of the primer and the sample or for determining nucleic acid or amino acid sequence of the divalent sialoside binding site of a hemagglutinin protein of influenza virus comprising the steps of; (a) isolating genomic nucleic acid of an influenza virus; and (b) sequencing a nucleic acid sequence encoding the cysteine 97 region, cysteine 139 region and the region of amino acids 220-226 as defined by the amino acid sequence of X31-hemaglutinin, wherein said method optionally comprises a further step of designing peptides for influenza vaccine development based on the sequencing results obtained in step (b) and optionally the use further including contacting the sample with a nucleic acid microarray, the nucleic acid microarray comprising one or more primers, each of said primers comprising a sequence of any one listed in Table 1 or sequences in FIGS. 17-19 , under conditions suitable for hybridizing the one or more primers and the sample;and detecting hybridization of the one or more primers and the sample, or optionally the nucleotide being part of a nucleic acid microarray comprising a primer, said primer comprising a sequence of any one of Table 1 or sequences in FIGS. 17-19 or a kit comprising a primer and/or nucleic acid as defined above and instructions for detecting antigenic compound according to claim 1.
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88. The method according to the claim 1, for identifying influenza virus in a biological sample, the method comprising:
- (a) contacting the biological sample with an antibody substance capable of binding antigenic compound according to claim 1; and
(b) detecting the binding between said antibody substance and antigenic compound in the sample, said binding indicating the presence and type of influenza virus in the sample.
- (a) contacting the biological sample with an antibody substance capable of binding antigenic compound according to claim 1; and
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97. The substance according to claim 10, wherein the substance is according to Formula
[PEP-(y)p-(S)q-(z)r-]nPO-
(SP1)wherein PO is an oligomeric or polymeric carrier structure, PEP is the peptide epitope sequence as defined for Peptide 1, Peptide 2 and Peptide 3, PO is preferably selected from the group consisting of;
a) solid phases, b) immunogenic and or oligomeric or polymeric carrier such as multiple antigen presenting (MAP) constructs, proteins such as KLH (keyhole limpet hemocyanin oligosaccharide or polysaccharide structure, n is an integer>
1 indicating the number of PEP groups covalently attached to the carrier PO, S is a spacer group, p, q and r are each 0 or 1, whereby at least one of p and r is different from 0, y and z are linking groups, at least one of y and z being a linking atom group also referred as “
chemoselective ligation group”
, in a preferred embodiment comprising at least one an O-hydroxylamine residue —
O—
NH—
or —
O—
N═
, with the nitrogen atom being linked to the OS and/or PO structure, respectively, and the other y and z, if present, is a chemoselective ligation group, with the proviso that when n is 1, the carrier structure is a monovalent immunogenic carrier.
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Specification