Antibodies to TGF-beta

Abstract

The present invention relates to antibody molecules, in particular antibody molecules that bind Transforming Growth Factor beta (TGFβ), and uses thereof. More particularly, the invention relates to antibody molecules that bind and preferably neutralise TGFβ1, TGFβ2 and TGFβ3, so-called “pan-specific” antibody molecules, and uses of such antibody molecules. Preferred embodiments within the present invention are antibody molecules, whether whole antibody (e.g. IgG, such as IgG1 or IgG4) or antibody fragments (e.g. scFv, Fab, dAb).

1 . An isolated specific binding member which binds to and neutralizes human TGFβ1, TGFβ2 and TGFβ3, comprising an antigen-binding domain of an antibody, wherein said antigen binding domain comprises a set of CDRs HCDR1, HCDR2 and HCDR3, and wherein said antigen binding domain utilizes a human VH1 family gene and wherein said HCDR3 has the amino acid sequence of SEQ ID NO: 25.  
     
     
         2 . The specific binding member according to  claim 1 , wherein the human VH1 family gene is a human VH1-2 gene.  
     
     
         3 . The specific binding member according to  claim 2 , where in the human VH1-2 gene is a DP-88 gene.  
     
     
         4 . The specific binding member according to any one of  claims 1  to  3 , wherein the antigen binding domain further comprises a set of CDRs LCDR1, LCDR2 and LCDR3, and wherein said antigen binding domain utilizes a human Vκ3 family gene and wherein said LCDR3 has the amino acid sequence of SEQ ID NO: 30.  
     
     
         5 . The specific binding member according to  claim 4  wherein the HCDR3 and LCDR3 are SEQ ID NO: 25 and SEQ ID NO: 30, respectively.  
     
     
         6 . The specific binding member according to  claim 4 , wherein the human Vκ3 family gene is a human Vκ DPK22 gene.  
     
     
         7 . The specific binding member according to  claim 1 , wherein the HCDR1, HCDR2 and HCDR3 of the VH domain are within a germline heavy chain framework.  
     
     
         8 . The specific binding member according to  claim 1 , wherein the HCDR1, HCDR2 and HCDR3 of the VH domain are within a framework that comprises up to 12 mutations from the germline amino acid sequence.  
     
     
         9 . The specific binding member according to  claim 4 , wherein the LCDR1, LCDR2 and LCDR3 of the Vκ domain are within a germline heavy chain framework.  
     
     
         10 . The specific binding member according to  claim 4 , wherein the LCDR1, LCDR2 and LCDR3 of the Vκ domain are within a framework that comprises up to 5 mutations from the germline Vκ amino acid sequence.  
     
     
         11 . An isolated specific binding member that binds to and neutralizes human TGFβ1, TGFβ2 and TGFβ3, comprising an antigen-binding domain of an antibody, wherein said antigen binding domain utilizes a human VH DP-88 gene and comprises an FR4 amino acid sequence comprising the amino acid sequence in SEQ ID NO: 31.  
     
     
         12 . The specific binding member according to  claim 11 , wherein said antigen binding domain utilizes a human VH DP-88 gene, and comprises a set of CDRs HCDR1, HCDR2 and HCDR3, wherein said HCDR3 has the amino acid sequence of SEQ ID NO: 25, and further comprises an FR4 amino acid sequence comprising the amino acid sequence in SEQ ID NO: 31.  
     
     
         13 . The specific binding member according to  claim 11 , wherein the antigen binding domain further utilizes a human Vκ3 family gene and a human Jκ5 gene.  
     
     
         14 . The specific binding member according to  claim 13 , wherein said antigen binding domain utilizing a human Vκ3 family gene and a human Jκ5 gene comprises a set of CDRs LCDR1, LCDR2 and LCDR3, and wherein said LCDR3 has the amino acid sequence of SEQ ID NO: 30.  
     
     
         15 . An isolated specific binding member which binds to and neutralises human TGFβ1, TGFβ2 and TGFβ3, comprising an antigen-binding domain of an antibody, wherein said antigen binding domain comprises the HCDR1 amino acid sequence of SEQ ID NO: 23, HCDR2 amino acid sequence of SEQ ID NO: 24, and the HCDR3 amino acid sequence of SEQ ID NO: 25.  
     
     
         16 . The isolated specific binding member according to  claim 15 , wherein the antigen-binding domain further comprises an antibody VL domain.  
     
     
         17 . The isolated specific binding member according to  claim 15 , wherein the antigen-binding domain comprises the LCDR1 amino acid sequence of SEQ ID NO: 28, LCDR2 amino acid sequence of SEQ ID NO: 29, and the LCDR3 amino acid sequence of SEQ ID NO: 30.  
     
     
         18 . The isolated specific binding member according to  claim 15  wherein HCDR1, HCDR2 and HCDR3 of the VH domain are within a germline heavy chain framework.  
     
     
         19 . The isolated specific binding member according to  claim 18 , wherein the germline heavy chain framework is a human VH1 family framework.  
     
     
         20 . The isolated specific binding member according to  claim 15 , wherein the HCDR1, HCDR2 and HCDR3 of the VH domain are within germline human heavy chain framework VH1 DP-88.  
     
     
         21 . The isolated specific binding member according to  claim 17 , wherein the LCDR1, LCDR2 and LCDR3 of the VL domain are within a germline light chain framework.  
     
     
         22 . The isolated specific binding member according to  claim 21 , wherein the germline light chain framework is a human Vκ3 family framework.  
     
     
         23 . The isolated specific binding member according to  claim 21 , wherein the antigen binding domain further utilizes a human Jκ5 gene.  
     
     
         24 . The isolated specific binding member according to  claim 22 , wherein the human Vκ3 family gene is a Vκ DPK22 gene.  
     
     
         25 . An isolated specific binding member comprising the PET1287A10 VH domain (SEQ ID NO: 22) with up to 5 mutations, or an antigen-binding portion thereof.  
     
     
         26 . The isolated specific binding member according to  claim 25  further comprising the PET1287A10 VL domain (SEQ ID NO: 27) with up to 5 mutations, or an antigen-binding portion thereof.  
     
     
         27 . An antibody comprising the PET 1287A10 VH domain (SEQ ID NO: 22) and the PET 1287A10 VL domain (SEQ ID NO: 27).  
     
     
         28 . The isolated specific binding member according to  claim 1  that comprises an scFv antibody molecule.  
     
     
         29 . The isolated specific binding member according to  claim 1 , that comprises an antibody constant region.  
     
     
         30 . The isolated specific binding member according to  claim 29  wherein the constant region is from an IgG4.  
     
     
         31 . A composition comprising a specific binding member according to  claim 1 .  
     
     
         32 . An isolated nucleic acid which comprises a nucleotide sequence encoding a specific binding member according to  claim 1 .  
     
     
         33 . A host cell transformed with the nucleic acid according to  claim 32 .  
     
     
         34 . A method of producing a specific binding member comprising culturing a host cell according to  claim 33  under conditions for production of said specific binding member and isolating and/or purifying said specific binding member.  
     
     
         35 . The method according to  claim 34  further comprising formulating the specific binding member or antibody VH or VL variable domain into a composition including at least one additional active component.  
     
     
         36 . A method of producing a specific binding member that specifically binds human TGFβ1, TGFβ2 and TGFβ3, which method comprises: 
 (a) providing a 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 comprise germ-line human framework VH1 DP-88 and either comprise a HCDR1, HCDR2 and/or HCDR3 to be replaced or lack a HCDR1, HCDR2 and/or HCDR3 encoding region;    (b) combining said starting nucleic acid or starting repertoire with donor nucleic acid or donor nucleic acids, wherein the donor nucleic acid encodes a potential HCDR or the donor nucleic acids encode potential HCDRs, such that said donor nucleic acid is or donor nucleic acids are inserted into the HCDR1, HCDR2 and/or HCDR3 region in the starting nucleic acid or starting repertoire, so as to provide a product repertoire of nucleic acids encoding VH domains;    (c) expressing the nucleic acids of said product repertoire to produce product VH domains;    (d) optionally combining said product VH domains with one or more VL domains;    (e) selecting a specific binding member for human TGFβ1, TGFβ2 and TGFβ3, which specific binding member comprises a product VH domain and optionally a VL domain; and    (f) recovering said specific binding member or nucleic acid encoding it.    
     
     
         37 . The method according to  claim 36 , wherein the donor nucleic acid or donor nucleic acids encode or are produced by mutation of the amino acid sequence of: 
 (a) HCDR1 of SEQ ID NO: 23;    (b) HCDR2 of SEQ ID NO: 24; and/or    (c) HCDR3 of SEQ ID NO: 25.    
     
     
         38 . The method according to  claim 36  or  37 , further comprising fusing the recovered specific binding member to an antibody constant region.  
     
     
         39 . The method according to  claim 36 , wherein the specific binding member is an scfv antibody molecule.  
     
     
         40 . The method according to  claim 36 , wherein the specific binding member is an Fab antibody molecule.  
     
     
         41 . The method according to  claim 36  wherein the specific binding member is a whole antibody.  
     
     
         42 . A method of treating a disease or disorder selected from the group consisting of a fibrotic disease, cancer, or an immune-mediated disease by administering a pharmaceutically effective amount of a composition according to  claim 31 .  
     
     
         43 . Use of a specific binding member according to  claim 1  in the manufacture of a medicament for treatment of a disease or disorder selected from the group consisting of fibrotic disease, cancer, or an immune-mediated disease.  
     
     
         44 . A method of inhibiting TGFβ1, TGFβ2 or TGFβ3 signalling comprising the step of contacting TGFβ1, TGFβ2 and TGFβ3 in vivo with a specific binding member according to  claim 1 .  
     
     
         45 . A method for inhibiting TGFβ1, 2 or 3-mediated fibronectin production comprising the step of contacting TGFβ1, TGFβ2 or TGFβ3 with a specific binding member according to  claim 1 .  
     
     
         46 . A method for inhibiting TGFβ1, 2 or 3-mediated VEGF production comprising the step of contacting TGFβ1, TGFβ2 or TGFβ3 with a specific binding member according to  claim 1 .  
     
     
         47 . A method for modulating cell proliferation selected from the group consisting of: 
 (a) reducing TGFβ1, 2 or 3-mediated inhibition of epithelial cell proliferation;    (b) reducing TGFβ1, 2 or 3-mediated inhibition of endothelial cell proliferation; and    (c) inhibiting TGFβ1, 2 or 3-mediated smooth muscle cell proliferation, comprising the step of contacting cells expressing TGFβ1, TGFβ2 or TGFβ3 with a specific binding member according to  claim 1 .    
     
     
         48 . A method for inhibiting cyclosporin-induced TGFβ1, 2 or 3 activity comprising the step of contacting TGFβ1, TGFβ2 or TGFβ3 with a specific binding member according to  claim 1 .  
     
     
         49 . A method for increasing NK cell activity comprising the step of contacting cells expressing TGFβ1, TGFβ2 or TGFβ3 with a specific binding member according to  claim 1 .  
     
     
         50 . A method for inhibiting TGFβ1, 2 or 3-mediated immunosuppression comprising the step of contacting cells expressing TGFβ1, TGFβ2 or TGFβ3 with a specific binding member according to  claim 1 .  
     
     
         51 . A method for inhibiting the growth of a TGFβ1, 2 or 3 expressing tumor comprising the step of contacting cells expressing TGFβ1, TGFβ2 or TGFβ3 with a specific binding member according to  claim 1 .  
     
     
         52 . An isolated specific binding member which specifically binds to and neutralizes TGFβ1, TGFβ2 and TGFβ3 comprising a germline heavy chain framework sequence from the human VH1 gene family.  
     
     
         53 . The isolated specific binding member according to  claim 52 , wherein the germline heavy chain framework sequence is from the human DP-88 VH1 gene family.  
     
     
         54 . An isolated specific binding member which specifically binds to and neutralizes TGFβ1, TGFβ2 and TGFβ3 comprising a germline light chain sequence from the human Vκ3 gene family.  
     
     
         55 . The isolated specific binding member according to  claim 54 , wherein the framework region for the light chain is from DPK-22.  
     
     
         56 . The isolated specific binding member according to  claim 29  wherein the constant region is from an IgG1.