Engineered antibody for inhibition of fibrosis

Abstract

A chimeric, humanized or single-chain antibody contains a light chain variable region containing the complementarity determining regions of SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3, and a heavy chain variable region containing the complementarity determining regions SEQ ID NO:5 and SEQ ID NO:6. The antibody or antibody fragment thereof is capable of binding the C-terminal telopeptide of the α2(I) chain of human collagen I, and is useful in the treatment of diseases or disorders associated with excessive collagen fibril.

What is claimed is: 
     
         1 . A chimeric, humanized or single-chain antibody which comprises a light chain variable region comprising complementarity determining regions comprising the amino acid sequences SEQ ID NO: 1, SEQ ID NO:2 and SEQ ID NO:3, and a heavy chain variable region comprising complementarity determining regions comprising the amino acid sequences SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6, or a fragment of said chimeric or humanized antibody, which antibody or antibody fragment binds the C-terminal telopeptide of the a2(I) chain of human collagen I. 
     
     
         2 . The antibody or antibody fragment according to  claim 1 , which is a chimeric antibody or fragment of a chimeric antibody, which fragment binds the C-terminal telopeptide of the a2(I) chain of human collagen. 
     
     
         3 . The chimeric antibody according to  claim 2 , which comprises an antibody light chain comprising a light variable region having the amino acid sequence shown in SEQ ID NO.T0, and an antibody heavy chain comprising a heavy chain variable region having the amino acid sequence SEQ ID NO: l 1, or a fragment of said chimeric antibody which binds the C-terminal telopeptide of the a2(I) chain of human collagen. 
     
     
         4 . The chimeric antibody according to  claim 3 , wherein said antibody light chain comprises a human antibody light chain constant region, and said antibody heavy chain comprises a human antibody heavy chain constant region, or a fragment of said chimeric antibody which binds the C-terminal telopeptide of the oc2(I) chain of human collagen. 
     
     
         5 . The chimeric antibody according to  claim 4 , wherein the human constant regions comprise IgG constant regions. 
     
     
         6 . The antibody or antibody fragment according to  claim 1 , which is a humanized antibody or fragment of a humanized antibody, which fragment binds the C-terminal telopeptide of the ct2(I) chain of human collagen. 
     
     
         7 . The humanized antibody according to  claim 6 , which comprises a human antibody framework region and/or a human antibody constant region, or a fragment of said humanized antibody which binds the C-terminal telopeptide of the 2(I) chain of human collagen. 
     
     
         8 . The humanized antibody according to  claim 6 , which comprises a human antibody constant region. 
     
     
         9 . The humanized antibody according to  claim 8 , wherein the constant region comprises an IgG constant region. 
     
     
         10 . The antibody according to  claim 1 , which is a single chain antibody. 
     
     
         11 . The single chain antibody according to  claim 10 , wherein the single chain antibody comprises a light chain variable region having the amino acid sequence shown in SEQ ID NO:9, SEQ ID NO: 10 or SEQ ID NO:26, and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: l 1. 
     
     
         12 . The single chain antibody according to  claim 11 , wherein the antibody further comprises a linker comprising a sequence of one or more amino acids linking said light chain variable region and said heavy chain variable region. 
     
     
         13 . The single chain antibody according to  claim 12 , wherein the linker comprises the amino acid sequence Gly-Gly-Ser or the amino acid sequence Gly-Gly-Gly-Gly-Ser (SEQ ID NO:29). 
     
     
         14 . The single chain antibody according to  claim 12 , wherein the linker connects the carboxy terminus of said light chain variable region to the amino terminus of said heavy chain variable region. 
     
     
         15 . The single chain antibody according to  claim 13 , wherein the linker comprises from two to twelve repeats of the amino acid sequence Gly-Gly-Ser, or from two to twelve repeats of the amino acid sequence Gly-Gly-Gly-Gly-Ser (SEQ ID NO:29). 
     
     
         16 . The single chain antibody according to  claim 15 , comprising the amino acid sequence shown in SEQ ID NO: 12. 
     
     
         17 . The single chain antibody according to  claim 12 , wherein the linker connects the carboxy terminus of said heavy chain variable region to the amino terminus of said light chain variable region. 
     
     
         18 . The single chain antibody according to  claim 17 , wherein the linker comprises from two to twelve repeats of the amino acid sequence Gly-Gly-Ser, or from two to twelve repeats of the amino acid sequence Gly-Gly-Gly-Gly-Ser (SEQ ID NO:29). 
     
     
         19 . The single chain antibody according to  claim 18 , comprising the amino acid sequence shown in SEQ ID NO: 16. 
     
     
         20 . A polynucleotide for expressing a chimeric antibody light chain, said polynucleotide comprising a first segment encoding an antibody light chain variable region having the amino acid sequence shown in SEQ ID NO: 10 and a second segment encoding a human antibody light chain constant region. 
     
     
         21 . The polynucleotide according to  claim 20 , wherein the first segment comprises the nucleotide sequence shown in SEQ ID NO:22. 
     
     
         22 . A polynucleotide for expressing a chimeric antibody heavy chain, said polynucleotide comprising a first segment encoding an antibody heavy chain variable region having the amino acid sequence shown in SEQ ID NO: l 1 and a second segment encoding a human antibody heavy chain constant region. 
     
     
         23 . The polynucleotide according to  claim 22 , wherein the first segment comprises the nucleotide sequence shown in SEQ ID NO:23. 
     
     
         24 . A polynucleotide for expressing a single chain antibody comprising a first segment encoding an antibody light chain variable region having the amino acid sequence shown in SEQ ID NO:9, SEQ ID NO: 10 or SEQ ID NO:26, and a second segment encoding an antibody heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 11. 
     
     
         25 . The polynucleotide of  claim 24  comprising a linker segment between said first and second segments, said linker segment encoding a linker comprising one or more amino acids. 
     
     
         26 . The polynucleotide according to  claim 25 , wherein the linker segment encodes a linker comprising from two to twelve repeats of the amino acid sequence Gly-Gly-Ser, or from two to twelve repeats of the amino acid sequence Gly-Gly-Gly-Gly-Ser (SEQ ID NO:29). 
     
     
         27 . The polynucleotide according to  claim 26  wherein the first DNA segment comprises the nucleotide sequence SEQ ID NO:22, SEQ ID NO:27 or SEQ ID NO:28, and the second DNA segment comprises the nucleotide sequence SEQ ID NO:23. 
     
     
         28 . The polynucleotide according to  claim 24 , encoding a single chain antibody comprising the amino acid sequence SEQ ID NO: 12. 
     
     
         29 . The polynucleotide according to  claim 28 , comprising the nucleotide sequence SEQ ID NO: 13. 
     
     
         30 . The polynucleotide according to  claim 24 , encoding a single chain antibody comprising the amino acid sequence SEQ ID NO: 16. 
     
     
         31 . The polynucleotide according to  claim 30 , comprising the nucleotide sequence SEQ ID NO: 17. 
     
     
         32 . A pharmaceutical composition comprising an antibody or antibody fragment according to  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         33 . A method of treating a disease or disorder associated with excessive collagen fibril formation in a subject, comprising administering to the subject an effective amount of an antibody or antibody fragment according to  claim 1 . 
     
     
         34 . The method according to  claim 33 , wherein the disease or disorder comprises scar formation. 
     
     
         35 . The method according to  claim 34 , wherein the disease or disorder comprises fibrosis. 
     
     
         36 . The method according to  claim 35 , wherein the fibrosis comprises pulmonary fibrosis, idiopathic pulmonary fibrosis, cirrhosis, endomyocardial fibrosis, mediastinal fibrosis, myelofibrosis, retroperitoneal fibrosis, progressive massive fibrosis, nephrogenic systemic fibrosis, Crohn's Disease, keloid formation, myocardial infarction, scleroderma/systemic sclerosis, arthrofibrosis, or adhesive capsulitis. 
     
     
         37 . The method according to  claim 35  wherein the fibrosis results from a surgical procedure. 
     
     
         38 . The method according to  claim 37  wherein the surgery is abdominal surgery, plastic surgery, glaucoma surgery or surgery for implantation of a medical implant or device.