PEPTIDE VACCINE FOR INFLUENZA VIRUS

• US 20110191867A1
• Filed: 10/24/2008
• Published: 08/04/2011
• Est. Priority Date: 10/26/2006
• Status: Abandoned Application

• Current Assignee: Glykos Finland Oy
• Original Assignee: Glykos Finland Oy
• Inventors: Aitio, Olli, Niemela, Ritva, Hiltunen, Jukka, Natunen, Jari, Helin, Jari
• Application Number:
  US12/739,796
• Publication Number:
  US 20110191867A1
• Time in Patent Office: Days
• US Class Current: 800/8
• CPC Class Codes: A61K 2039/64 characterised by the architecture of the carrier-antigen complex, e.g. repetition of carrier-antigen units
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: A61K 39/12 Viral antigens
• CPC Class Codes: A61K 39/145 Orthomyxoviridae, e.g. influenza virus
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 31/16 for influenza or rhinoviruses
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C07K 5/0815 with the first amino acid being basic
• CPC Class Codes: C07K 5/0821 with the first amino acid being heterocyclic, e.g. His, Pro, Trp
• CPC Class Codes: C12N 2760/16122 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2760/16134 Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• CPC Class Codes: G01N 2333/11 Orthomyxoviridae, e.g. influenza virus
• CPC Class Codes: G01N 33/56983 Viruses
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

Abstract

The invention provides peptide epitopes for use in the prevention and/or treatment of influenza or for the development of such treatment or vaccine against influenza. The invention also relates to a 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, and to conjugates containing one or more such peptide epitopes. The peptide epitopes of the invention are cyclic peptides comprising a 7-mer peptide derived from H1, H3 or H5 hemagglutinin of influenza virus. The 7-mer peptide has a sequence corresponding to the loop sequence at positions 220-226 of X31-hemagglutinin.

Claims

1-33. -33. (canceled)

34. Peptide conjugate according to Formula

[PEP-(y)_p-(S)_q-(z)_r-]_nPO

wherein PEP is a peptide epitope;

n is an integer ≧

1 indicating the number of PEP groups covalently attached to carrier PO;

S is a spacer group;

y and z are linking groups so that at least y or z is a linking atom group;

p, q and r are independently 0 or 1 so that at least p or r is 1;

PO is an oligomeric or polymeric carrier structure;

wherein said PEP is a cyclic peptide comprising a 7-mer peptide derived from H1, H3, or H5 hemagglutinin of influenza virus, said peptide having a sequence the location of which in said hemagglutinin corresponds to the loop sequence at positions 220-226 of X31-hemagglutinin.

35. The conjugate according to claim 34, wherein said 7-mer peptide has a loop structure conformationally similar to the loop sequence at positions 220-226 of X31-hemagglutinin.

36. The conjugate according to claim 34, wherein said cyclic peptide comprises the H1, H3 and H5 concensus sequence according to Formula

(C)_nRX₁X₂VX₃ (SEQ ID NO:

1), wherein

X₁ is P or S,

X₂ is R or K, and

X₃ is R or N,

(C) is optional cysteine of the cyclic peptide and n is 0 or 1 indicating its presence or absence or

the cyclic peptide comprises H3 type sequence

37. The conjugate according to claim 34, wherein the sequence of said cyclic peptide comprises H1 type sequence KVR, H3 type sequence RVR or H5 type sequence KVN.

38. The conjugate according to claim 34, wherein the sequence of said cyclic peptide comprises H1 type sequence KVR, H3 type sequence RVR or H5 type sequence KVN with cysteines added to both ends of the 7-mer peptide resulting in the following structures:

39. The conjugate according to claim 34, wherein the sequence of said cyclic peptide comprises RPRVRNI (SEQ ID NO:

5), RPRIRNI (SEQ ID NO;

6), RSKVNGQ (SEQ ID NO;

7), or RPKVRDQ (SEQ ID NO;

8).

40. The conjugate according to claim 38, wherein the carboxyl terminal cysteine is further linked to a spacer, additional hemagglutinin peptide or glycine amide.

41. The conjugate according to claim 34, wherein z is selected form the group:

a chemoselective ligation group or biotin or equivalent ligand capable of specific strong non-covalent interaction.

42. The conjugate according to claim 34, wherein PO is selected from the group consisting of:

solid phases, immunogenic and/or oligomeric or polymeric carrier such as multiple antigen presenting (MAP) constructs, proteins such as KLH (keyhole limpet hemocyanin), and oligosaccharide or polysaccharide structures.

43. The conjugate according to claim 34, wherein said z or y is selected from a group consisting of:

a linking atom group formed from sulphur atom of a cysteine residue, preferably linked to maleimide or analogous structure or to a sulphur of cysteine in the matrix or the linking group is a strong non-covalent interaction formed by binding of a ligand to a protein, preferably biotin binding to an avidin protein, and an O-hydroxylamine residue —

O—

NH—

or —

O—

N═

, with the nitrogen atom being linked to the OS or PO structure, respectively.

45. A method for evaluating the potential of a chemical entity to bind to the conjugate according to claim 34 comprising the steps of:

(i) contacting said chemical entity with said conjugate or peptide under conditions that allow said chemical entity to bind said conjugate or peptide; and

(ii) detecting the presence of a complex of said chemical entity and said conjugate or peptide.

46. The method according to claim 45, wherein said chemical entity is an antibody.

47. The method according to claim 45, wherein the use of the method is selected from the group consisting of:

vaccine development, selection of antibodies from a library of antibodies;

an ex vivo or in vivo immunization method, screening binding agents from a library;

screening antibodies, wherein said chemical entity is an antibody and said peptide is contacted with a sample of whole blood, plasma or serum, an in vitro immunoassay or in vitro selection of an antibody library such as phage display antibody library, a method of detecting the presence of an antibody in a biological sample.

48. The method according to claim 47, wherein the use of the method is producing a peptide vaccine against influenza, the method further comprising steps of:

administering the conjugate according to claim 34 to an animal; and

monitoring the animal in order to detect immune response against the conjugate.

49. The conjugate according to claim 34 as a part of a vaccine composition.

50. The conjugate according to claim 49 further comprising a linear peptide having the same sequence as the cyclic peptide.

51. The method according to claim 45 for identifying influenza virus in a biological sample, the method further comprising:

(a) contacting the biological sample with an antibody substance capable of binding the conjugate according to claim 34; and

(b) detecting the binding between said antibody substance and influenza virus or part thereof in the sample, said binding indicating the presence and type of influenza virus in the sample.

52. The method according to claim 45 for screening variants of a 7-mer peptide derived from H1, H3, or H5 hemagglutinin of influenza virus, said peptide having a sequence the location of which in said hemagglutinin corresponds to the loop sequence at positions 220-226 of X31-hemagglutinin;

the method comprising the steps of;

(a) acquiring sequence data of hemagglutinin from data banks or by sequencing influenza virus genomes;

(b) obtaining candidate peptide sequences by comparing the sequences to known sequences from the same location;

(c) preparing a cyclic peptide or a conjugate as defined in claim 34;

(d) screening antibodies binding to the peptide or conjugate obtained in step (c).

53. The method according to claim 52, wherein the method further involves a step of search of any of the peptide epitopes 1-3, more preferably the peptide 3, of an hemagglutinin from database comprising human genome coded peptide sequences and selection of peptides, which are not expected to cause immune reaction against the human (or animal) subject.

44. Cyclic peptide of 7-12 amino acids comprising the sequence selected from the group consisting of RPRVRNI (SEQ ID NO:

5), RPRIRNI (SEQ ID NO;

6), or RSKVNGQ (SEQ ID NO;

7), CRPRVRNIC (SEQ ID NO;

9), CRPRIRNIC (SEQ ID NO;

10), or CRSKVNGQC (SEQ ID NO;

11);

or any of the peptides ID NO 9-11 or cyclic peptide CRPKVRDQC (SEQ ID NO;

14), wherein the carboxyl terminal cysteine of said peptide is further linked to a spacer, additional hemagglutinin peptide or glycine amide.

54. A method for evaluating the potential of a chemical entity to bind to the cyclic peptide according to claim 44 comprising the steps of:

(i) contacting said chemical entity with said conjugate or peptide under conditions that allow said chemical entity to bind said conjugate or peptide; and

(ii) detecting the presence of a complex of said chemical entity and said conjugate or peptide.

55. The method according to claim 45 for identifying influenza virus in a biological sample, the method further comprising:

(a) contacting the biological sample with an antibody substance capable of binding the cyclic epitope according to claim 44; and

(b) detecting the binding between said antibody substance and influenza virus or part thereof in the sample, said binding indicating the presence and type of influenza virus in the sample.