Polypeptides binding ADAMTS5, MMP13 and aggrecan
The present invention relates to polypeptides binding Aggrecan as well as ADAMTS5 and/or MMP13, more in particular to polypeptides that comprise or essentially consist of immunoglobulins binding Aggrecan as well as immunoglobulins binding ADAMTS5 and/or immunoglobulins binding MMP13 (also referred to herein as “polypeptides of the invention”, and “immunoglobulin(s) of the invention”, respectively). The invention also relates to constructs comprising such immunoglobulins, such as immunoglobulin single variable domains (ISVDs) or polypeptides as well as nucleic acids encoding such immunoglobulins or polypeptides (also referred to herein as “nucleic acid(s) of the invention”; to methods for preparing such immunoglobulins, polypeptides and constructs; to host cells expressing or capable of expressing such immunoglobulins or polypeptides; to compositions, and in particular to pharmaceutical compositions, that comprise such immunoglobulins, polypeptides, constructs, nucleic acids and/or host cells; and to uses of immunoglobulins, polypeptides, constructs, nucleic acids, host cells and/or compositions, in particular for prophylactic and/or therapeutic purposes, such as the prophylactic and/or therapeutic purposes mentioned herein. Other aspects, embodiments, advantages and applications of the invention will become clear from the further description herein.
1. A polypeptide comprising at least 3 ISVDs, in which a first ISVD specifically binds matrix metalloproteinase 13 (MMP13), a second ISVD specifically binds ADAMTS5 and a third ISVD specifically binds Aggrecan;
wherein said ISVD specifically binding MMP13 comprises 3 complementarity determining regions, wherein the complementarity determining regions are CDR1 to CDR3, in which
(i) CDR1 comprises SEQ ID NO: 8;
(ii) CDR2 comprises SEQ ID NO: 10; and
(iii) CDR3 comprises SEQ ID NO: 12;
wherein said ISVD specifically binding ADAMTS5 comprises 3 complementarity determining regions, wherein the complementarity determining regions are CDR1 to CDR3, in which
(i) CDR1 comprises SEQ ID NO: 14;
(ii) CDR2 comprises SEQ ID NO: 16; and
(iii) CDR3 comprises SEQ ID NO: 18; and
wherein said ISVD specifically binding Aggrecan comprises 3 complementarity determining regions, wherein the complementarity determining regions are CDR1 to CDR3, in which
(i) CDR1 comprises (a) SEQ ID NO: 19, or (b) an amino acid sequence according to SEQ ID NO: 19 that has amino acid substitutions at positions 7 and/or 9, wherein the amino acid substitutions are:
N at position 7 in SEQ ID NO: 19 changed to S; and/or
V at position 9 changed to M;
(ii) CDR2 comprises (a) SEQ ID NO: 21, or (b) an amino acid sequence according to SEQ ID NO: 21 that has amino acid substitutions at positions 1, 3, 4, 8, 9, and/or combinations thereof, wherein the amino acid substitutions are:
T at position 1 in SEQ ID NO: 21 changed to A;
S at position 3 in SEQ ID NO: 21 changed to R;
S at position 4 in SEQ ID NO: 21 changed to T;
A at position 8 in SEQ ID NO: 21 changed to T; and/or
N at position 9 in SEQ ID NO: 21 changed to D; and
(iii) CDR3 comprises (a) SEQ ID NO: 23, or (b) an amino acid sequence according to SEQ ID NO: 23 that has amino acid substitutions at positions 4 and/or 8, wherein the amino acid substitutions are:
H at position 4 in SEQ ID NO: 23 changed to R; and/or
V at position 8 in SEQ ID NO: 23 changed to D.
2. The polypeptide according to claim 1 , wherein said ISVD specifically binding MMP13 comprises or consists of SEQ ID NO: 2.
3. The polypeptide according to claim 1 , wherein said ISVD specifically binding ADAMTS5 comprises or consists of SEQ ID NO: 3.
4. The polypeptide according to claim 1 , wherein said ISVD specifically binding Aggrecan comprises or consists of SEQ ID NO: 4.
5. The polypeptide according to claim 1 , wherein said ISVDs are linked to each other via a linker selected from the group consisting of SEQ ID NOs: 24 to 40.
6. The polypeptide according to claim 1 , in which said polypeptide comprises a first ISVD specifically binding MMP13, a second ISVD specifically binding ADAMTS5, a third ISVD specifically binding Aggrecan and the polypeptide further comprises a fourth ISVD specifically binding Aggrecan, wherein the fourth ISVD has the same CDR1, CDR2 and CDR3 as defined in claim 1 for the third ISVD binding Aggrecan.
7. The polypeptide according to claim 6 , wherein said polypeptide comprises or consists of SEQ ID NO: 1 or 62, or comprises or consists of a polypeptide that has at least 95% sequence identity to SEQ ID NO: 1 or 62.
8. A pharmaceutical composition comprising the polypeptide according to claim 1 .
9. A nucleic acid encoding the polypeptide according to claim 1 .
10. An expression vector comprising the nucleic acid according to claim 9 .
11. A host or host cell comprising the nucleic acid according to claim 9 .
12. A method for producing a, comprising the steps of:
a) expressing, in a suitable host cell, host organism or suitable expression system, the nucleic acid according to claim 9 ; optionally followed by
b) isolating and/or purifying the polypeptide.
13. A method of treating a disease or disorder in an individual, the method comprising administering the polypeptide according to claim 1 to said individual in an amount effective to treat the disease or disorder, wherein the disease or disorder is selected from the group consisting of arthropathies and chondrodystrophies, arthritic disease, osteoarthritis, rheumatoid arthritis, gouty arthritis, psoriatic arthritis, traumatic rupture or detachment, achondroplasia, costochondritis, Spondyloepimetaphyseal dysplasia, spinal disc herniation, lumbar disk degeneration disease, degenerative joint disease, relapsing polychondritis, osteochondritis dissecans, aggrecanopathies, NASH, chronic periodontitis and abdominal aortic aneurysms.