IL218091A - Polypeptide or polypeptide construct comprising at least one single domain antibody directed against von willebrand factor, a composition comprising it, a nucleic acid encoding it and uses thereof in the preparation of a medicament for prevention and/or alleviation of conditions of platelet-mediated aggregation - Google Patents

Description

- 172 - 218091/2 A polypeptide or polypeptide construct comprising at least one single domain antibody directed against von Willebrand Factor (vWF), wherein the at least one single domain antibody corresponds to a sequence represented by any of SEQ ID NO:s 5, 3 or 7, or to: i. an homologous sequence of any of SEQ ID NO:s 5, 3 or 7 with a sequence identity of more than 70% with the parent sequence; or ii. a functional portion of any of SEQ ID NO:s 5, 3 or 7 that maintains the interaction with vWF with affinity of 1 x 10"6 M or better; or iii. a functional portion of any of SEQ ID NO:s 5, 3 or 7 that comprises a partial deletion of the complete amino acid sequence and still maintains the binding site(s) and protein domain(s) necessary for the binding of and interaction with vWF; and, at least one single domain antibody directed against serum albumin (SA), wherein the at least one single domain antibody corresponds to a sequence represented by any of SEQ ID NO:s 16 to 19, or to: i. an homologous sequence of any of SEQ ID NO:s 16 to 19 with a sequence identity of more than 70% with the parent sequence.

The polypeptide or polypeptide construct according to claim 1 , in which at least one single domain antibody is a VHH domain.

The polypeptide or polypeptide construct according to claims 1 to 2, in which at least one single domain antibody is a VHH domain comprising an amino acid at position 45 according to the Kabat numbering that is selected from the group consisting of glycine, alanine, valine, leucine, isoleucine, proline, phenylalanine, tyrosine, tryptophan, methionine, serine, threonine, asparagine, and glutamine.

The polypeptide or polypeptide construct according to claims 1 to 3, in which at least one single domain antibody is a VHH domain comprising an amino acid at position 103 according to the Kabat numbering selected from the group consisting of arginine, serine or an uncharged residue, optionally glycine.

The polypeptide or polypeptide construct according to claims 1 to 4, in which at least one single domain antibody is a VHH domain that is obtained by immunising a camel and obtaining hybridoma's therefrom, or by cloning a library of single domain antibodies and subsequently selecting the VHH using phage display. - 173 - 218091/2 . The polypeptide or polypeptide construct according to claims 1 to 5, in which at least one single domain antibody is humanized.

. The polypeptide or polypeptide construct according to claims 1 to 6, in which at least one single domain antibody is a humanized VHH domain.

. The polypeptide or polypeptide construct according to claims 1 to 7, in which at least one single domain antibody is humanized by replacing one or more of the Camelidae amino acids by their human counterparts as found in a human consensus sequence.

. The polypeptide or polypeptide construct according to claims 1 to 8, in which at least one single domain antibody is humanized by replacing any of the following residues either alone or in combination: FR1 positions 1 , 5, 28 and 30, the hallmark amino acids at FR2 positions 37, 44, 45 and 47, FR3 positions 74, 75, 76, 83, 84, 93 and 94 and FR4 positions 103, 104, 108 and 111 , wherein the numbering of the positions is according to the Kabat numbering.

. The polypeptide or polypeptide construct according to claims 1 to 9, in which the C- terminal end of the first single domain antibody is linked to the N-terminal end of the next single domain antibody. 11. The composition comprising a polypeptide or polypeptide construct according to claims 1 to 10 and a pharmaceutically acceptable vehicle. 12. The composition comprising a polypeptide or polypeptide construct according to claims 1 to 10 adapted to the chosen route of administration, wherein the route of administration is orally or parenterally, intra-nasally by inhalation, intravenous, intramuscular, topical or subcutaneous routes; and a pharmaceutically acceptable vehicle. 13. The nucleic acid encoding a polypeptide or polypeptide construct according to claims 1 to 10. 14. The use of a polypeptide or polypeptide construct according to claims 1 to 10 for the preparation of a medicament for the prevention, treatment and/or alleviation of conditions of platelet-mediated aggregation wherein said conditions are any of the formation of a non-occlusive thrombus, the formation of an occlusive thrombus, arterial thrombus formation, acute coronary occlusion, restenosis, restenosis after PCTA or stenting, thrombus formation in stenosed arteries, hyperplasia after angioplasty, atherectomy or arterial stenting, occlusive syndrome in a vascular system or lack of patency of diseased arteries.