Polypeptides comprising immunoglobulin single variable domains targeting il-13 and OX40L
The present disclosure provides a novel type of drug for treating a subject suffering from an inflammatory disease. Specifically, the disclosure provides polypeptides comprising at least three immunoglobulin single variable domains (ISVDs), characterized in that at least one ISV binds to OX40L and at least two ISVDs bind to IL-13. The present disclosure also provides nucleic acids, vectors and compositions.
1 . A polypeptide comprising at least three immunoglobulin single variable domains (ISVDs), optionally linked via one or more peptidic linkers, wherein each of said ISVDs comprises three complementarity determining regions (CDR1 to CDR3, respectively); and wherein:
a) a first ISVD that binds to OX40L and comprises:
i. a CDR1 which has the amino acid sequence of SEQ ID NO: 6;
ii. a CDR2 which has the amino acid sequence of SEQ ID NO: 10; and
iii. a CDR3 which has the amino acid sequence of SEQ ID NO: 14;
b) a second ISVD that binds to IL-13 and comprises:
iv. a CDR1 which has the amino acid sequence of SEQ ID NO: 7;
v. a CDR2 which has the amino acid sequence of SEQ ID NO: 11; and
vi. a CDR3 which has the amino acid sequence of SEQ ID NO: 15; and
c) a third ISVD that binds to IL-13 and comprises:
vii. a CDR1 which has the amino acid sequence of SEQ ID NO: 9;
viii. a CDR2 which has the amino acid sequence of SEQ ID NO: 13; and
ix. a CDR3 which has the amino acid sequence of SEQ ID NO: 17.
2 . The polypeptide according to claim 1 , wherein:
a) the amino acid sequence of said first ISVD has a sequence identity of more than 90% with SEQ ID NO: 2;
b) the amino acid sequence of said second ISVD has a sequence identity of more than 90% with SEQ ID NO: 3; and
c) the amino acid sequence of said third ISVD has a sequence identity of more than 90% identity with SEQ ID NO: 5.
3 . The polypeptide according to claim 1 , wherein:
a) said first ISVD has the amino acid sequence of SEQ ID NO: 2;
b) said second ISVD has the amino acid sequence of SEQ ID NO: 3; and
c) said third ISVD has the amino acid sequence of SEQ ID NO: 5.
4 . The polypeptide according to claim 1 , wherein said polypeptide further comprises one or more other groups, residues, moieties or binding units, optionally linked via one or more peptidic linkers, in which said one or more other groups, residues, moieties or binding units provide the polypeptide with increased half-life, compared to the corresponding polypeptide without said one or more other groups, residues, moieties or binding units.
5 . The polypeptide according to claim 4 , in which said one or more other groups, residues, moieties or binding units that provide the polypeptide with increased half-life is chosen from the group consisting of a polyethylene glycol molecule, serum proteins or fragments thereof, binding units that can bind to serum proteins, an Fc portion, and small proteins or peptides that can bind to serum proteins.
6 . The polypeptide according to claim 4 , in which said one or more other groups, residues, moieties or binding units that provide the polypeptide with increased half-life is an ISVD that binds to human serum albumin and comprises: a CDR1 having the amino acid sequence of SEQ ID NO:8; a CDR2 having the amino acid sequence of SEQ ID NO: 12; and a CDR3 having the amino acid sequence of SEQ ID NO: 16.
7 . The polypeptide according to claim 1 , further comprising a fourth ISVD, wherein the fourth ISVD comprises: i. a CDR1 which has the amino acid sequence of SEQ ID NO: 8 or has 2 or 1 amino acid difference(s) with SEQ ID NO: 8; ii. a CDR2 which has the amino acid sequence of SEQ ID NO: 12 or has 2 or 1 amino acid difference(s) with SEQ ID NO: 12; and iii. a CDR3 which has the amino acid sequence of SEQ ID NO: 16 or has 2 or 1 amino acid difference(s) with SEQ ID NO: 16.
8 . The polypeptide according to claim 7 , wherein the amino acid sequence of said fourth ISVD has a sequence identity of more than 90% with SEQ ID NO: 4.
9 . The polypeptide according to claim 7 , wherein said fourth ISVD has the amino acid sequence of SEQ ID NO: 4.
10 . The polypeptide of claim 7 , wherein the polypeptide comprises, from N-terminally to C-terminally, the first ISVD, the second ISVD, the fourth ISVD, and the third ISVD.
11 . The polypeptide according to claim 1 , wherein the amino acid sequence of the polypeptide has a sequence identity of more than 90% with SEQ ID NO: 1.
12 . The polypeptide according to claim 1 , wherein the amino acid sequence of the polypeptide has the amino acid sequence of SEQ ID NO: 1.
13 . An isolated nucleic acid comprising a nucleotide sequence that encodes a polypeptide according to claim 1 .
14 . An isolated host cell comprising a nucleic acid according to claim 13 .
15 . A method for producing a polypeptide, said method comprising the steps of: a) expressing a nucleic acid according to claim 13 ; optionally followed by: b) isolating and/or purifying the polypeptide.
16 . A composition comprising at least one polypeptide according to claim 1 , optionally, wherein the composition is a pharmaceutical composition which further comprises: at least one pharmaceutically acceptable carrier, diluent or excipient and/or adjuvant; and/or one or more further pharmaceutically active polypeptides and/or compounds.
17 . A method for the treatment of an inflammatory disease, wherein said inflammatory disease is selected from the group consisting of asthma and atopic dermatitis, said method comprising administering, to a subject in need thereof, a pharmaceutically active amount of a polypeptide according to claim 1 .
18 . The method according to claim 17 , wherein the inflammatory disease is selected from the group consisting of asthma and atopic dermatitis.
19 . The polypeptide of claim 1 , wherein the polypeptide comprises, N-terminally to C-terminally, the first ISVD, the second ISVD, and the third ISVD.
20 . An immunoglobulin single domain (ISVD) that specifically binds to human OX40L, wherein said ISVD comprises three complementarity determining regions (CDR1 to CDR3, respectively); and wherein: CDR1 has the amino acid sequence of SEQ ID NO: 6; CDR2 has the amino acid sequence of SEQ ID NO: 10; and CDR3 has the amino acid sequence of SEQ ID NO: 14.