Compounds

Abstract

The present invention relates to binding members, especially antibody molecules, for CXCL13. The binding members are useful for the treatment of disorders associated with CXCL13, including arthritic disorders such as rheumatoid arthritis.

1 . An isolated binding member for CXCL13, wherein the binding member inhibits binding of CXCL13 to CXCR5, and wherein the binding member competes for binding to CXCL13 with an antibody scFv molecule having amino acid sequence SEQ ID NO: 11. 
     
     
         2 . A binding member according to  claim 1 , wherein the binding member comprises an HCDR3 amino acid sequence SEQ ID NO: 5. 
     
     
         3 . An isolated binding member according to  claim 1  or  claim 2  comprising a set of antibody 1 CDRs: HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, defined wherein:
 HCDR1 has amino acid sequence SEQ ID NO: 3;   HCDR2 has amino acid sequence SEQ ID NO: 4;   HCDR3 has amino acid sequence SEQ ID NO: 5;   LCDR1 has amino acid sequence SEQ ID NO: 8;   LCDR2 has amino acid sequence SEQ ID NO: 9; and   LCDR3 has amino acid sequence SEQ ID NO: 10;   or comprising the antibody 1 set of CDRs with one or more amino acid substitutions, deletions or insertions.   
     
     
         4 . An isolated binding member for CXCL13, wherein the binding member inhibits binding of CXCL13 to CXCR5 and wherein the binding member comprises a set of antibody 1 CDRs: HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, defined wherein:
 HCDR1 has amino acid sequence SEQ ID NO: 3;   HCDR2 has amino acid sequence SEQ ID NO: 4;   HCDR3 has amino acid sequence SEQ ID NO: 5;   LCDR1 has amino acid sequence SEQ ID NO: 8;   LCDR2 has amino acid sequence SEQ ID NO: 9; and   LCDR3 has amino acid sequence SEQ ID NO: 10;   or comprises the antibody 1 set of CDRs with one or more amino acid substitutions, deletions or insertions.   
     
     
         5 . A binding member according to  claim 3  or  claim 4  comprising the set of antibody 1 CDRs or comprising the set of antibody 1 CDRs with 1 or 2 substitutions. 
     
     
         6 . A binding member according to  claim 3  or  4  comprising the antibody 1 set of CDRs. 
     
     
         7 . A binding member according to any of the preceding claims, wherein the binding member comprises an antibody molecule comprising an antibody VH domain and an antibody VL domain, wherein the antibody molecule comprises a set of CDRs: HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3, wherein the VH domain comprises HCDR1, HCDR2, HCDR3 and a framework and the VL domain comprises LCDR1, LCDR2, LCDR3 and a framework. 
     
     
         8 . A binding member according to  claim 7 , wherein the antibody molecule is an scFv. 
     
     
         9 . A binding member according to  claim 7 , wherein the antibody molecule comprises an antibody constant region. 
     
     
         10 . A binding member according to  claim 9 , wherein the antibody molecule is an IgG1. 
     
     
         11 . A binding member according to any of  claims 7  to  10 , wherein the antibody molecule comprises framework regions of human germline gene segment sequences. 
     
     
         12 . A binding member according to any of  claims 7  to  11 , wherein the antibody VH domain comprises human germline framework VH3-23. 
     
     
         13 . A binding member according to any of  claims 7  to  12 , wherein the antibody VH domain has an amino acid sequence that has at least 90% sequence identity with SEQ ID NO: 2. 
     
     
         14 . A binding member according to any of  claims 7  to  11 , wherein the antibody VH domain amino acid sequence is SEQ ID NO: 2. 
     
     
         15 . A binding member according to any of  claims 11  to  14 , wherein the antibody VL domain comprises human germline framework VK1 L12. 
     
     
         16 . A binding member according to any of  claims 11  to  15 , wherein the antibody molecule comprises a VL domain having an amino acid sequence that has at least 90% sequence identity with SEQ ID NO: 7. 
     
     
         17 . A binding member according to any of  claims 11  to  14 , wherein the VL domain amino acid sequence is SEQ ID NO: 7. 
     
     
         18 . A binding member according to any of  claims 7  to  10 , wherein the antibody VH domain amino acid sequence is SEQ ID NO: 2; and the antibody VL domain amino acid sequence is SEQ ID NO: 7. 
     
     
         19 . A binding member according to any of the preceding claims, wherein potency of the binding member for neutralising the binding of human CXCL13 to CXCR5 differs by not more than 10-fold from potency of the binding member for neutralising cynomolgus CXCL13 binding to CXCR5, wherein
 potency of the binding member for neutralising the binding of human CXCL13 to CXCR5 is represented by IC 50  as measured in a cAMP assay with a human CXCL13 concentration of not more than 2 nM, and wherein   potency of the binding member for neutralising the binding of cynomolgus CXCL13 to CXCR5 is represented by IC 50  as measured in a cAMP assay with a final concentration of 2 nM cynomolgus CXCL13.   
     
     
         20 . A binding member according to  claim 19 , wherein the potency for neutralising the binding of human CXCL13 binding to CXCR5 differs by not more than 5-fold from the potency for neutralising the binding of cynomolgus CXCL13 to CXCR5. 
     
     
         21 . A binding member according to any of the preceding claims, wherein the binding member has an IC 50  of not more than 12 nM as measured in a human CXCL13 cAMP assay with a final concentration of 2 nM CXCL13. 
     
     
         22 . A binding member according to any of the preceding claims, wherein the binding member has an IC 50  of not more than 40 nM as measured in a human CXCL13 calcium release assay with a final concentration of 100 nM CXCL13. 
     
     
         23 . A binding member according to any of the preceding claims, wherein the binding member has an IC 50  of not more than 12 nM in a human CXCL13 B cell chemotaxis assay with a final concentration of human CXCL13 which gives an approximately ED80 response, and a non-primary B cell line. 
     
     
         24 . A binding member according to any of the preceding claims, wherein the binding member binds human CXCL13 with an affinity of less than 400 pM as measured using surface plasmon resonance. 
     
     
         25 . A binding member according to any of the preceding claims, wherein the binding member binds cynomolgus CXCL13 with an affinity of less than 400 pM as measured using surface plasmon resonance. 
     
     
         26 . A binding member according to any of the preceding claims, wherein the binding member binds an epitope of human CXCL-13 and wherein said epitope includes at least one residue of the sequence Ile-Leu-Pro-Arg-Gly-Asn-Gly-Cys-Pro-Arg-Lys-Glu (SEQ ID NO: 20) at positions 31-42 of mature human IL-17A. 
     
     
         27 . An isolated VH domain of an antibody molecule according to any of  claims 7  to  26 . 
     
     
         28 . An isolated VL domain of an antibody molecule according to any of  claims 7  to  26 . 
     
     
         29 . A composition comprising an isolated binding member according to any of  claims 1  to  26  and a pharmaceutically acceptable excipient. 
     
     
         30 . A composition comprising an isolated binding member according to any of  claims 1  to  26  for use in a method of treatment of the human or animal body by surgery or therapy. 
     
     
         31 . A composition according to  claim 30  for use in inhibiting binding of CXCL13 to CXCR5. 
     
     
         32 . Use of an isolated binding member according to any of  claims 1  to  26  for the manufacture of a medicament for inhibiting binding of CXCL13 to CXCR5. 
     
     
         33 . A composition according to  claim 31  or use according to  claim 32 , for use in inhibiting aberrant formation and/or development of lymphoid follicles. 
     
     
         34 . A composition according to  claim 31  or use according to  claim 32 , for use in inhibiting destruction and/or remodelling of bone and/or cartilage. 
     
     
         35 . A composition according to  claim 31  or use according to  claim 32 , for use in inhibiting chemotaxis of lymphocytes and/or proliferation of lymphomas. 
     
     
         36 . A composition according to  claim 31  or use according to  claim 32 , for use in treating rheumatoid arthritis, osteoarthritis, Sjogrens syndrome, multiple sclerosis, myasthenia gravis, systemic lupus erythmatosis, autoimmune thyroid disease, Lyme Neuroborreliosis, HIV infection and/or leukaemia. 
     
     
         37 . A composition according to  claim 31  or use according to  claim 32 , for use in treating rheumatoid arthritis. 
     
     
         38 . A method of treating a disorder associated with aberrant CXCL13 expression and/or CXCL13 activity in an individual, comprising administering a binding member according to any of  claims 1  to  26  to the individual. 
     
     
         39 . A method according to  claim 38  wherein the CXCL13 activity is binding to CXCR5. 
     
     
         40 . A method according to  claim 39  comprising inhibiting aberrant formation and/or development of lymphoid follicles in the individual. 
     
     
         41 . A method according to  claim 39  comprising inhibiting destruction and/or remodelling of bone and/or cartilage in the individual. 
     
     
         42 . A method according to  claim 39  comprising inhibiting chemotaxis of lymphocytes and/or proliferation of lymphomas in the individual. 
     
     
         43 . A method according to any of  claims 38  to  39 , wherein the disorder is rheumatoid arthritis, osteoarthritis, Sjogrens syndrome, multiple sclerosis, myasthenia gravis, systemic lupus erythmatosis, autoimmune thyroid disease, Lyme Neuroborreliosis, HIV infection and/or leukaemia. 
     
     
         44 . A method according to  claim 43 , wherein the disorder is rheumatoid arthritis. 
     
     
         45 . An isolated nucleic acid molecule comprising a nucleotide sequence encoding a binding member, or an isolated VH or VL domain of a binding member, according to any of  claims 1  to  28 . 
     
     
         46 . A host cell in vitro transformed with nucleic acid according to  claim 45 . 
     
     
         47 . A method of producing a binding member or an antibody VH or VL domain, comprising culturing host cells according to  claim 46  under conditions for production of the binding member or antibody VH or VL domain. 
     
     
         48 . A method according to  claim 47 , further comprising isolating and/or purifying the binding member, VH domain or VL domain. 
     
     
         49 . A method according to  claim 47  or  claim 48 , further comprising formulating the binding member, VH domain or VL domain into a composition comprising at least one additional component. 
     
     
         50 . A method for producing an antibody antigen-binding domain for CXCL13, the method comprising
 providing, by way of addition, deletion, substitution or insertion of one or more amino acids in the amino acid sequence of a parent VH domain comprising HCDR1, HCDR2 and HCDR3, wherein the parent VH domain HCDR1, HCDR2 and HCDR3 are a set of antibody 1 HCDRs, a VH domain which is an amino acid sequence variant of the parent VH domain, and optionally combining the VH domain thus provided with one or more VL domains to provide one or more VH/VL combinations; and   testing said VH domain which is an amino acid sequence variant of the parent VH domain or the VH/VL combination or combinations to identify an antibody antigen binding domain for CXCL13.   
     
     
         51 . A method according to  claim 50 , wherein the parent VH domain is a VH domain according to  claim 27 . 
     
     
         52 . A method according to  claim 50  or  claim 51  wherein said one or more VL domains is provided by way of addition, deletion, substitution or insertion of one or more amino acids in the amino acid sequence of a parent VL domain comprising LCDR1, LCDR2 and LCDR3, wherein the parent VL domain LCDR1, LCDR2 and LCDR3 are a VL set of antibody 1 CDRs, producing one or more VL domains each of which is an amino acid sequence variant of the parent VL domain. 
     
     
         53 . A method according to  claim 52  wherein the parent VL domain is a VL domain according to  claim 28 . 
     
     
         54 . A method according to any one of  claims 50  to  53 , wherein said VH domain which is an amino acid sequence variant of the parent VH domain is provided by CDR mutagenesis. 
     
     
         55 . A method according to any one of  claims 50  to  53  further comprising producing the antibody antigen-binding domain as a component of an IgG, scFv or Fab antibody molecule. 
     
     
         56 . A method for producing a binding member that binds CXCL13, which method comprises:
 providing starting nucleic acid encoding a VH domain or a starting repertoire of nucleic acids each encoding a VH domain, wherein the VH domain or VH domains either comprise a HCDR1, HCDR2 and/or HCDR3 to be replaced or lack a HCDR1, HCDR2 and/or HCDR3 encoding region;   combining said starting nucleic acid or starting repertoire with donor nucleic acid or donor nucleic acids encoding or produced by mutation of the amino acid sequence of an HCDR1, HCDR2, and/or HCDR3 of antibody 1, such that said donor nucleic acid is or donor nucleic acids are inserted into the CDR1, CDR2 and/or CDR3 region in the starting nucleic acid or starting repertoire, so as to provide a product repertoire of nucleic acids encoding VH domains;   expressing the nucleic acids of said product repertoire to produce product VH domains;   optionally combining said product VH domains with one or more VL domains;   selecting a binding member for CXCL13, which binding member comprises a product VH domain and optionally a VL domain; and recovering said binding member or nucleic acid encoding it.   
     
     
         57 . A method according to  claim 56  wherein the donor nucleic acids are produced by mutation of said HCDR1 and/or HCDR2. 
     
     
         58 . A method according to  claim 56  wherein the donor nucleic acid is produced by mutation of HCDR3. 
     
     
         59 . A method according to  claim 56 , comprising providing the donor nucleic acid by random mutation of nucleic acid. 
     
     
         60 . A method according to any one of  claims 56  to  59 , further comprising attaching a product VH domain that is comprised within the recovered binding member to an antibody constant region. 
     
     
         61 . A method according to any one of  claims 56  to  60  comprising providing an IgG, scFv or Fab antibody molecule comprising the product VH domain and a VL domain. 
     
     
         62 . A method according to any one of  claims 50  to  61 , further comprising testing the antibody antigen-binding domain or binding member that binds CXCL13 for ability to neutralize CXCL13. 
     
     
         63 . A method according to  claim 62 , wherein a binding member that comprises an antibody molecule that binds and neutralizes CXCL13 is obtained. 
     
     
         64 . A method according to  claim 63 , wherein the antibody molecule is an scFv. 
     
     
         65 . A method according to  claim 63 , wherein the antibody molecule is an IgG. 
     
     
         66 . A method for producing an antibody molecule composition, comprising obtaining an antibody molecule using a method according to any of  claims 50  to  65 , and formulating the antibody molecule into a composition comprising at least one additional component.