Amino acid sequences directed against human respiratory syncytial virus (HRSV) and polypeptides comprising the same for the prevention and/or treatment of respiratory tract infections

Patent metadata

Field	Value
Publication number	US-9644022-B2
Application number	US-201013512100-A
Country	US
Kind code	B2
Filing date	Nov 30, 2010
Priority date	Nov 30, 2009
Publication date	May 9, 2017
Grant date	May 9, 2017

Abstract

Amino acid sequences are provided that are directed against/and or that can specifically bind protein F of hRSV, as well as to compounds or constructs, and in particular proteins and polypeptides, that comprise or essentially consist of one or more such amino acid sequences. The amino acid sequences, polypeptides and therapeutic compounds and compositions provided by the invention show an improved stability, less immunogenicity and/or improved affinity and/or avidity for protein F of hRSV. The invention also relates to the uses of such amino acid sequences, polypeptides, compounds or constructs for prophylactic and/or therapeutic purposes.

First claim

The invention claimed is:

1. A monovalent polypeptide that specifically binds protein F of hRSV comprising SEQ ID NO: 16 wherein one or more amino acid residues have been mutated selected from the group consisting of:
(a) Glu1Asp;
(b) Ala14Pro Lys83Arg and Gln108Leu;
(c) Asp16Gly;
(d) Asp54Glu; and
(e) Gly55Ala,
wherein said positions are determined according to Kabat numbering.

2. A polypeptide that specifically binds protein F of hRSV comprising a VHH, a humanized VHH, a camelized VH or a domain antibody with:
a) the amino acid sequence of any one of SEQ ID NO: 33, 34, 35, 36, 37, 38, 39 or 40; or
b) an amino acid sequence having no more than 3, preferably no more than 2, more preferably no more than 1 amino acid difference with the amino acid sequence of any one of SEQ ID NO: 33, 34, 35, 36 37, 38, 39, or 40, provided that:
i) the amino acid sequence of the VHH, humanized VHH, camelized VH or the domain antibody has Proline (Pro, P) at position 14, Arginine (Arg, R) at position 83 and Leucine (Leu, L) at position 108, wherein said positions are determined according to Kabat numbering;
ii) the VHH, humanized VHH, camelized VH or the domain antibody comprises the following CDR sequences:
CDR1 is SYAMG (amino acids 31-35 of SEQ ID NO:33);
CDR2 is AISWSDGSTYYADSVKG (amino acids 50-66 of SEQ ID NO:33), AISWSEGSTYYADSVKG (amino acids 50-66 of SEQ ID NO:37), or AISWSDASTYYADSVKG (amino acids 50-66 of SEQ ID NO:39); and
CDR3 is DLTSTNPGSYIYIWAYDY (amino acids 99-116 of SEQ ID NO:33); and

iii) the polypeptide binds protein F of hRSV with the same, about the same, or a higher affinity, and/or the polypeptide has the same, about the same, or a higher potency compared to the polypeptide without the 3, 2 or 1 amino acid difference.

3. The polypeptide according to claim 2 , comprising
a) the amino acid sequence of any one of SEQ ID NO: 35, 36, 37, 38, 39 or 40; or
b) an amino acid sequence having no more than 3, preferably no more than 2, more preferably no more than 1 amino acid difference with the amino acid sequence of any one of SEQ ID NO: 35, 36, 37, 38, 39, or 40, provided that:
i) the amino acid sequence of the VHH, humanized VHH, camelized VH or the domain antibody has
Proline (Pro, P) at position 14, Arginine (Arg, R) at position 83, and Leucine (Leu, L) at position 108; and
Glycine (Gly, G) at position 16, Glutamic acid (Glu, E) at position 54 and/or Alanine (Ala, A) at position 55,
wherein said positions are determined according to Kabat numbering; and

ii) the polypeptide binds protein F of hRSV with the same, about the same, or a higher affinity and/or polypeptide has the same, about the same, or a higher potency compared to the polypeptide without the 3, 2 or 1 amino acid difference.

4. A polypeptide that specifically binds protein F of hRSV, comprising a VHH, a humanized VHH, a camelized VH or a domain antibody with:
a) the amino acid sequence of any one of SEQ ID NO: 34, 36, 38, 40, or 51; or
b) an amino acid sequence having no more than 3, preferably no more than 2, more preferably no more than 1 amino acid difference with the amino acid sequence of any one of SEQ ID NO: 34, 36, 38, or 40, provided that:
i) the amino acid sequence of the VHH, humanized VHH, camelized VH or the domain antibody has Aspartic acid (Asp, D) at position 1, wherein said position is determined according to Kabat numbering;
ii) the VHH, humanized VHH, camelized VH or the domain antibody comprises the following CDR sequences:
CDR1 is SYAMG (amino acids 31-35 of SEQ ID NO:33);
CDR2 is AISWSDGSTYYADSVKG (amino acids 50-66 of SEQ ID NO:33), AISWSEGSTYYADSVKG (amino acids 50-66 of SEQ ID NO:37), or AISWSDASTYYADSVKG (amino acids 50-66 of SEQ ID NO:39); and
CDR3 is DLTSTNPGSYIYIWAYDY (amino acids 99-116 of SEQ ID NO:33); and

iii) the polypeptide binds protein F of hRSV with the same, about the same, or a higher affinity and/or the polypeptide has the same, about the same, or a higher potency compared to the polypeptide without the 3, 2 or 1 amino acid difference.

5. The monovalent polypeptide according to claim 1 , that comprises or consists of the amino acid sequence of any one of SEQ ID NO: 33, 34, 35, 36, 37, 38, 39, 40, or 51.

6. A polypeptide that comprises one or more monovalent polypeptides according to claim 1 , and optionally further comprises one or more other polypeptides, optionally linked via one or more peptidic linkers.

7. The polypeptide according to claim 6 , wherein the polypeptide is a multivalent polypeptide that comprises at least two monovalent polypeptides.

8. The multivalent polypeptide according to claim 7 , wherein the multivalent polypeptide comprises at least three monovalent polypeptides.

9. The multivalent polypeptide according to claim 7 , wherein the multivalent polypeptide has aspartic acid (Asp, D) at position 1.

10. The multivalent polypeptide according to claim 7 , wherein the multivalent polypeptide comprises or consists of two monovalent polypeptides.

11. The multivalent polypeptide according to claim 10 , wherein the multivalent polypeptide has aspartic acid (Asp, D) at position 1.

12. The multivalent polypeptide according to claim 7 , wherein the multivalent polypeptide comprises or consists of three polypeptides.

13. The polypeptide according to claim 12 , wherein the multivalent polypeptide has aspartic acid (Asp, D) at position 1.

14. The multivalent polypeptide according to claim 12 wherein the multivalent polypeptide that specifically binds protein F of hRSV, comprises three VHHs, humanized VHHs, camelized VHs or single domain antibodies with:
a) the amino acid sequence of any one of SEQ ID NO: 41, 42, 43, 44, 45, 46, 47, 48, or 49; or
b) an amino acid sequence having no more than 3, preferably no more than 2, more preferably no more than 1 amino acid difference with the amino acid sequence of any one of SEQ ID NO: 41, 42, 43, 44, 45, 46, 47, 48, or 49, provided that:
i) the VHHs, humanized VHHs, camelized VHs or domain antibodies have Proline (Pro, P) at position 14, Arginine (Arg, R) at position 83 and Leucine (Leu, L) at position 108, wherein said positions are determined according to Kabat numbering; and
ii) the VHHs, humanized VHHs, camelized VHs or domain antibodies comprise the following CDR sequences:
CDR1 is SYAMG (amino acids 31-35 of SEQ ID NO:33);
CDR2 is AISWSDGSTYYADSVKG (amino acids 50-66 of SEQ ID NO:33), AISWSEGSTYYADSVKG (amino acids 50-66 of SEQ ID NO:37), or AISWSDASTYYADSVKG (amino acids 50-66 of SEQ ID NO:39); and
CDR3 is DLTSTNPGSYIYIWAYDY (amino acids 99-116 of SEQ ID NO:33); and

iii) the multivalent polypeptide binds protein F of hRSV with the same, about the same, or a higher affinity and/or the multivalent polypeptide has the same, about the same, or a higher potency compared to the polypeptide without the 3, 2 or 1 amino acid difference.

15. The multivalent polypeptide according to claim 14 , wherein the multivalent polypeptide that specifically binds protein F of hRSV comprises,
a) the amino acid sequence of any one of SEQ ID NO: 42, 43, 44, 47, 48, or 49;
b) an amino acid sequence having no more than 3, preferably no more than 2, more preferably no more than 1 amino acid difference with the amino acid sequence of any one of SEQ ID NO: 42, 43, 44, 47, 48, or 49, provided that:
i) the VHHs, humanized VHHs, camelized VHs or domain antibodies have
Proline (Pro, P) at position 14, Arginine (Arg, R) at position 83 and Leucine (Leu, L) at position 108; and
Glycine (Gly, G) at position 16, Glutamic acid (Glu, E) at position 54 and/or Alanine (Ala, A) at position 55,
wherein said positions are determined according

Assignees

• Depla Erik
• Stortelers Catelijne
• Ablynx Nv

Inventors

• Depla Erik
• Stortelers Catelijne

Classifications

• A61P31/14

  for RNA viruses · CPC title

• A61P29/00

  Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID] · CPC title

• C07K2317/76

  Antagonist effect on antigen, e.g. neutralization or inhibition of binding · CPC title

• C07K2317/22

  from camelids, e.g. camel, llama or dromedary · CPC title

• C07K2317/569

  Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody® · CPC title