Anti-edb antibodies and antibody-drug conjugates

Abstract

The present invention provides antibodies and antibody-drug conjugates that bind to the extra domain B splice variant of fibronectin 1 and methods for preparing and using the same.

What is claimed is: 
     
         1 . An antibody-drug conjugate comprising (a) an antibody, or antigen binding fragment thereof, that binds to extra domain B of fibronectin, (b) a linker and (c) a drug. 
     
     
         2 . The antibody-drug conjugate of  claim 1 , wherein the antibody, or antigen binding fragment, comprises a heavy chain variable region comprising three CDRs comprising SEQ ID NOs: 3, 5 and 7, and a light chain variable region comprising three CDRs comprising SEQ ID NOs: 12, 13 and 14. 
     
     
         3 . The antibody-drug conjugate of  claim 1  or  2 , wherein the antibody, or antigen binding fragment, comprises a heavy chain variable region comprising SEQ ID NO: 1 or 21, and a light chain variable region comprising SEQ ID NO: 10. 
     
     
         4 . The antibody-drug conjugate of any one of  claims 1 - 3 , wherein the antibody, or antigen binding fragment, comprises
 a heavy chain variable region comprising SEQ ID NO: 1 and a light chain variable region comprising SEQ ID NO: 10; or   a heavy chain variable region comprising SEQ ID NO: 21 and a light chain variable region comprising SEQ ID NO: 10.   
     
     
         5 . The antibody-drug conjugate of any one of  claims 1 - 4 , wherein the antibody, or antigen binding fragment, comprises a heavy chain comprising SEQ ID NO: 8, 17, 19, 23, 25, 27 or 29, and a light chain comprising SEQ ID NO: 15 or 31. 
     
     
         6 . The antibody-drug conjugate of any one of  claims 1 - 5 , wherein the antibody, or antigen binding fragment, comprises
 a heavy chain comprising SEQ ID NO: 8 and a light chain comprising SEQ ID NO: 15;   a heavy chain comprising SEQ ID NO: 8 and a light chain comprising SEQ ID NO: 31;   a heavy chain comprising SEQ ID NO: 17 and a light chain comprising SEQ ID NO: 15;   a heavy chain comprising SEQ ID NO: 17 and a light chain comprising SEQ ID NO: 31;   a heavy chain comprising SEQ ID NO: 19 and a light chain comprising SEQ ID NO: 15;   a heavy chain comprising SEQ ID NO: 19 and a light chain comprising SEQ ID NO: 31;   a heavy chain comprising SEQ ID NO: 23 and a light chain comprising SEQ ID NO: 15;   a heavy chain comprising SEQ ID NO: 23 and a light chain comprising SEQ ID NO: 31;   a heavy chain comprising SEQ ID NO: 25 and a light chain comprising SEQ ID NO: 15;   a heavy chain comprising SEQ ID NO: 25 and a light chain comprising SEQ ID NO: 31;   a heavy chain comprising SEQ ID NO: 27 and a light chain comprising SEQ ID NO: 15;   a heavy chain comprising SEQ ID NO: 27 and a light chain comprising SEQ ID NO: 31;   a heavy chain comprising SEQ ID NO: 29 and a light chain comprising SEQ ID NO: 15; or   a heavy chain comprising SEQ ID NO: 29 and a light chain comprising SEQ ID NO: 31.   
     
     
         7 . The antibody-drug conjugate of  claim 1 , wherein the antibody, or antigen binding fragment, comprises a heavy chain and/or light chain constant region comprising an engineered cysteine residue for site-specific conjugation. 
     
     
         8 . The antibody-drug conjugate of  claim 7 , wherein the heavy chain constant region comprises an engineered cysteine residue at position 290 (K290C), according to the numbering of the EU index of Kabat. 
     
     
         9 . The antibody-drug conjugate of  claim 7 , wherein the light chain constant region comprises an engineered cysteine residue at position 183 (κK183C), according to the numbering of Kabat. 
     
     
         10 . The antibody-drug conjugate of  claim 1 , wherein the heavy chain constant region comprises an engineered cysteine residue at position 290 (K2900), according to the numbering of the EU index of Kabat, and wherein the light chain constant region comprises an engineered cysteine residue at position 183 (κK183C), according to the numbering of Kabat. 
     
     
         11 . The antibody-drug conjugate of  claim 1 , wherein the antibody, or antigen binding fragment, comprises a heavy chain constant region comprising an engineered glutamine-containing tag inserted in the antibody or replaces one or more endogenous amino acids in the antibody. 
     
     
         12 . The antibody-drug conjugate of  claim 11 , wherein the engineered glutamine-containing tag is inserted in the antibody at position E294-N297. 
     
     
         13 . The antibody-drug conjugate of  claim 12 , wherein the glutamine-containing tag comprises an amino acid sequence LLQG (SEQ ID NO: 40). 
     
     
         14 . The antibody-drug conjugate of  claim 11 , wherein the heavy chain constant region further comprises a lysine (K) substituting an arginine (R) at position 222 (K222R), according to the numbering of the EU index of Kabat. 
     
     
         15 . The antibody-drug conjugate of  claim 1 , wherein the antibody, or antigen binding fragment, comprises a heavy chain variable region comprising a lysine (K) substituting an arginine (R) at position 94 (K94R), according to the numbering of Kabat. 
     
     
         16 . The antibody-drug conjugate of any one of  claims 1 - 15 , wherein the linker is a cleavable linker. 
     
     
         17 . The antibody-drug conjugate of  claim 16 , wherein the cleavable linker is selected from the group consisting of vc, diS, diS-C 2 OCO and AcLys-vc. 
     
     
         18 . The antibody-drug conjugate of any one of  claims 1 - 17 , wherein the drug is a cytotoxic agent. 
     
     
         19 . The antibody-drug conjugate of  claim 18 , wherein the cytotoxic agent is an auristatin. 
     
     
         20 . The antibody-drug conjugate of  claim 19 , wherein the auristatin is selected from the group consisting of 0101, 1569, 9411 and 4574. 
     
     
         21 . The antibody-drug conjugate of any one of  claims 1 - 18 , wherein the cytotoxic agent is a CPI dimer. 
     
     
         22 . The antibody-drug conjugate of  claim 21 , wherein the CPI dimer is CPI-8314 or CPI-0326. 
     
     
         23 . An antibody-drug conjugate comprising (a) an antibody, or antigen binding fragment thereof, comprising a heavy chain variable region comprising three CDRs comprising SEQ ID NOs: 3, 5 and 7, and a light chain variable region comprising three CDRs comprising SEQ ID NOs: 12, 13 and 14, (b) a vc linker and (c) a 0101 drug. 
     
     
         24 . An antibody-drug conjugate comprising (a) an antibody, or antigen binding fragment thereof, comprising a heavy chain variable region comprising SEQ ID NO: 21 and a light chain variable region comprising SEQ ID NO: 10; (b) a vc linker and (c) a 0101 drug. 
     
     
         25 . An antibody-drug conjugate comprising (a) an antibody, or antigen binding fragment thereof, comprising a heavy chain comprising SEQ ID NO: 25 and a light chain comprising SEQ ID NO: 31; (b) a vc linker and (c) a 0101 drug. 
     
     
         26 . A pharmaceutical composition comprising the antibody-drug conjugate of any of  claims 1 - 25  and a pharmaceutically acceptable carrier. 
     
     
         27 . A composition comprising a plurality of an antibody-drug conjugates of any one of  claims 1 - 25 , and optionally a pharmaceutical carrier, wherein the composition has an average DAR of ranging from 3 to 5. 
     
     
         28 . A composition comprising a plurality of an antibody-drug conjugates of any one of  claims 1 - 25 , and optionally a pharmaceutical carrier, wherein the composition has an average DAR of ranging from 1 to 3. 
     
     
         29 . A nucleic acid encoding a heavy chain or a light chain of the antibody of any one of  claims 1 - 25 . 
     
     
         30 . A nucleic acid of any of SEQ ID NOs: 9, 18, 20, 24, 26, 28 or 30 encoding a heavy chain or any of SEQ ID NOs: 16 or 32 encoding a light chain. 
     
     
         31 . A vector comprising the nucleic acid of  claim 29  or  30 . 
     
     
         32 . A host cell comprising the nucleic acid of  claim 29  or  30 . 
     
     
         33 . A process for producing an antibody-drug conjugate of any one of  claims 1 - 25  comprising:
 (a) linking the linker to the drug; 
 (b) conjugating the linker and drug to the antibody; and 
 (c) purifying the antibody-drug conjugate. 
 
     
     
         34 . The process of  claim 33 , wherein the conjugating is site-specific on one or more engineered cysteine residue and/or engineered glutamine residues on the antibody. 
     
     
         35 . A method of treating an EDB+ FN-expressing disorder or disease, comprising administering an effective amount of a composition comprising an antibody-drug conjugate of any one of  claims 1 - 25  to a subject in need thereof. 
     
     
         36 . The method of  claim 35 , wherein EDB+ FN-expressing disorder or disease is cancer. 
     
     
         37 . The method of  claim 36 , wherein the cancer is a solid tumor or blood cancer. 
     
     
         38 . The method of  claim 37 , wherein the solid tumor is thyroid cancer, sarcoma, breast cancer, pancreatic cancer, glioblastoma, gallbladder cancer, kidney cancer, skin cancer, uterine cancer, mesothelioma, colorectal cancer, head and neck cancer, ovarian cancer, bladder cancer, testicular cancer, prostate cancer, liver cancer, endocrine cancer, thymus cancer, brain cancer, adrenal cancer, eye cancer cervical cancer and lung cancer. 
     
     
         39 . The method of  claim 36 , wherein the blood cancer is leukemia, lymphoma or myeloma. 
     
     
         40 . Use of the antibody-drug conjugate of any one of  claims 1 - 25 , in the manufacture of a medicament for the treatment of an EDB+ FN-expressing disorder or disease in a subject. 
     
     
         41 . The use according to  claim 40 , wherein the EDB+ FN-expressing disorder or disease is cancer. 
     
     
         42 . The use according to  claim 41 , wherein the cancer is a solid tumor or blood cancer. 
     
     
         43 . The use according to  claim 42 , wherein the solid tumor is thyroid cancer, sarcoma, breast cancer, pancreatic cancer, glioblastoma, gallbladder cancer, kidney cancer, skin cancer, uterine cancer, mesothelioma, colorectal cancer, head and neck cancer, ovarian cancer, bladder cancer, testicular cancer, prostate cancer, liver cancer, endocrine cancer, thymus cancer, brain cancer, adrenal cancer, eye cancer cervical cancer and lung cancer. 
     
     
         44 . The use according to  claim 43 , wherein the blood cancer is leukemia, lymphoma or myeloma.