Anti-endoglin antibodies and uses thereof

Abstract

The present application relates to compositions of humanized and deimmunized anti-endoglin antibodies and antigen-binding fragments thereof. One aspect relates to antibodies having one or more modifications in at least one amino acid residue of at least one of the framework regions of the variable heavy chain, the variable light chain or both. Another aspect relates to anti-endoglin antibodies which inhibit or treat fibrosis.

What is claimed is: 
     
         1 . A composition comprising an endoglin binding antibody or binding fragment thereof and a pharmaceutically acceptable excipient for use in treating or inhibiting fibrosis, wherein the antibody or the antigen binding fragment thereof comprises a heavy chain variable region and a light chain variable region wherein:
 the heavy chain variable region comprises an amino acid sequence as set forth in SEQ ID NO:89 optionally comprising one or more modifications selected from the group consisting of a substitution of glycine (G) by alanine (A) or serine (S) at position 49; a substitution of alanine (A) by isoleucine (I) at position 51; a substitution of lysine (K) by arginine (R) or asparagine (Q) at position 52b; a substitution of leucine (L) by valine (V) at position 78 utilising the Kabat number system; and   wherein the light chain variable region comprises an amino acid sequence as set forth in SEQ ID NO:89 optionally comprising one or more modifications selected from the group consisting of a substitution of methionine (M) by leucine (L) at position 4; a substitution of alanine (A) by valine (V) at position 19; a substitution of threonine (T) by serine (S) at position 22; a substitution of alanine (A) by isoleucine (I) at position 48; and a substitution of threonine (T) by serine(S) at position 51 utilising the Kabat number system;   and a pharmaceutically acceptable excipient, whereby fibrosis is treated or inhibited.   
     
     
         2 . The composition of  claim 1 , wherein the antibody, or antigen-binding fragment thereof, comprises a heavy chain variable region having an amino acid sequence set forth as SEQ ID NO: 88, 89, 90, 91 or 92; and a light chain variable region having an amino acid sequence set forth as SEQ ID NO: 93, 94, 95, 96, 97, 100, 102, or 103. 
     
     
         3 . A composition comprising an endoglin binding antibody or binding fragment thereof and a pharmaceutically acceptable excipient for use in treating or inhibiting fibrosis, wherein the antibody or the antigen binding fragment thereof comprises a heavy chain variable region having an amino acid sequence set forth as SEQ ID NO: 89 and a light chain variable region having an amino acid sequence set forth as SEQ ID NO: 93 and a pharmaceutically acceptable excipient, whereby fibrosis is treated or inhibited. 
     
     
         4 . The composition of any one of  claims 1 - 3 , wherein the antibody or binding fragment thereof is useful for treatment of liver fibrosis. 
     
     
         5 . The composition of  claim 4 , wherein liver fibrosis is selected from the group consisting of cirrhosis and associated conditions such as chronic viral hepatitis, non-alcoholic fatty liver disease (NAFLD), alcoholic steatohepatitis (ASH), non-alcoholic steatohepatitis (NASH), primary biliary cirrhosis (PBC), biliary cirrhosis, autoimmune hepatitis). 
     
     
         6 . The composition of any one of  claims 1 - 3 , wherein the antibody or binding fragment thereof is useful for treatment of kidney fibrosis. 
     
     
         7 . The composition of  claim 6 , wherein kidney fibrosis is caused by chronic kidney disease, metabolic syndrome, vesicoureteral reflux, tubulointerstitial renal fibrosis, diabetes, glomerulonephritis or glomerular nephritis (GN), focal segmental glomerulosclerosis and membranous glomerulonephritis, or mesangiocapillary GN. 
     
     
         8 . The composition of any one of  claims 1 - 3 , wherein the antibody or binding fragment thereof is useful for treatment of pulmonary fibrosis. 
     
     
         9 . The composition of  claim 8 , wherein pulmonary fibrosis is idiopathic pulmonary fibrosis (IPF) or cryptogenic fibrosing alveolitis, chronic fibrosing interstitial pneumonia, interstitial lung disease (ILD), and diffuse parenchymal lung disease (DPLD), idiopathic interstitial pneumonia or acute respiratory distress syndrome (ARDS). 
     
     
         10 . The composition of any one of  claims 1 - 3 , wherein the antibody or binding fragment thereof is useful for treatment of intestinal fibrosis. 
     
     
         11 . The composition of any one of  claims 1 - 3 , wherein the antibody or binding fragment thereof is useful for treatment of ocular fibrosis. 
     
     
         12 . The composition of any one of  claims 1 - 3 , wherein the antibody or binding fragment thereof is useful for treatment of fibrosis of joints. 
     
     
         13 . A method of treating or inhibiting fibrosis in a subject in need thereof, comprising administering to the subject a composition comprising an antibody, or antigen-binding fragment thereof, comprising a heavy chain variable region having an amino acid sequence set forth as SEQ ID NO: 89 and a light chain variable region having an amino acid sequence set forth as SEQ ID NO: 93 and a pharmaceutically acceptable excipient, whereby fibrosis is treated or inhibited. 
     
     
         14 . A method of treating or inhibiting fibrosis in a subject in need thereof, comprising administering to the subject a composition comprising an antibody, or antigen-binding fragment thereof, that binds endoglin, comprising a heavy chain variable region having an amino acid sequence set forth as SEQ ID NO: 89 and a light chain variable region having an amino acid sequence set forth as SEQ ID NO: 93, wherein:
 said heavy chain variable region further comprises one or more modifications selected from the group consisting of a substitution of glycine (G) by alanine (A) or serine (S) at position 49; a substitution of alanine (A) by isoleucine (I) at position 51; a substitution of lysine (K) by arginine (R) or asparagine (Q) at position 52b; a substitution of leucine (L) by valine (V) at position 78 utilizing the Kabat numbering system; and   the light chain variable region further comprises one or more modifications selected from the group consisting of a substitution of methionine (M) by leucine (L) at position 4; a substitution of alanine (A) by valine (V) at position 19; a substitution of threonine (T) by serine (S) at position 22; a substitution of alanine (A) by isoleucine (I) at position 48; and a substitution of threonine (T) by serine (S) at position 51 utilizing the Kabat numbering system;   and a pharmaceutically acceptable excipient, whereby fibrosis is treated or inhibited.   
     
     
         15 . The method of  claim 14 , wherein the antibody, or antigen-binding fragment thereof, comprises a heavy chain variable region having an amino acid sequence set forth as SEQ ID NO: 88, 89, 90, 91 or 92; and a light chain variable region having an amino acid sequence set forth as SEQ ID NO: 93, 94, 95, 96, 97, 100, 102, or 103. 
     
     
         16 . The method of any one of  claims 13 - 15 , wherein the antigen-binding fragment is a Fab fragment, a Fab′ fragment, a F(ab′)2 fragment, an Fv fragment, an scFv fragment, a single chain binding polypeptide, a Fd fragment, a variable heavy chain, a variable light chain or a dAb fragment. 
     
     
         17 . The method of any one of  claims 13 - 16 , wherein the antibody, or antigen-binding fragment thereof, is further labeled with a therapeutic label. 
     
     
         18 . The method of any one of  claims 13 - 17 , wherein fibrosis is liver fibrosis. 
     
     
         19 . The method of  claim 18 , wherein liver fibrosis is selected from the group consisting of cirrhosis and associated conditions such as chronic viral hepatitis, non-alcoholic fatty liver disease (NAFLD), alcoholic steatohepatitis (ASH), non-alcoholic steatohepatitis (NASH), primary biliary cirrhosis (PBC), biliary cirrhosis, autoimmune hepatitis). 
     
     
         20 . The method of  claim 18 , wherein liver fibrosis is caused by a chronic insult to the liver from a parasite or a virus. 
     
     
         21 . The method of  claim 20 , wherein parasite or virus is Hepatitis B Virus (HBV), Hepatitis B Virus (HCV), Human Immunodeficiency Virus (HIV), or schistosomiasis. 
     
     
         22 . The method of any one of  claims 13 - 17 , wherein fibrosis is kidney fibrosis. 
     
     
         23 . The method of  claim 22 , wherein kidney fibrosis is caused by chronic kidney disease, metabolic syndrome, vesicoureteral reflux, tubulointerstitial renal fibrosis, diabetes, glomerulonephritis or glomerular nephritis (GN), focal segmental glomerulosclerosis and membranous glomerulonephritis, or mesangiocapillary GN. 
     
     
         24 . The method of any one of  claims 13 - 17 , wherein fibrosis is pulmonary fibrosis. 
     
     
         25 . The method of  claim 24 , wherein pulmonary fibrosis is idiopathic pulmonary fibrosis (IPF) or cryptogenic fibrosing alveolitis, chronic fibrosing interstitial pneumonia, interstitial lung disease (ILD), and diffuse parenchymal lung disease (DPLD), idiopathic interstitial pneumonia or acute respiratory distress syndrome (ARDS). 
     
     
         26 . The method of any one of  claims 13 - 25 , further comprising administering one or more fibrosis inhibitors. 
     
     
         27 . The method of  claim 26 , wherein the anti-endoglin antibody, or antigen-binding fragment and one or more fibrosis inhibitors are administered at the same site. 
     
     
         28 . The method of  claim 26 , wherein the anti-endoglin antibody, or antigen-binding fragment and one or more fibrosis inhibitors are administered at different sites. 
     
     
         29 . The method of  claim 26 , wherein the anti-endoglin antibody, or antigen-binding fragment and one or more fibrosis inhibitors are administered sequentially. 
     
     
         30 . The method of  claim 26 , wherein the anti-endoglin antibody, or antigen-binding fragment and one or more fibrosis inhibitors are administered concurrently. 
     
     
         31 . The method of any one of  claims 13 - 30 , wherein the antibody or antigen-binding fragment thereof is administered in an amount of about 0.01 mg/kg, about 0.05 mg/kg, about 0.1 mg/kg, about 0.5 mg/kg, about 1 mg/kg, about 5 mg/kg, about 10 mg/kg, about 20 mg/kg, about 30 mg/kg, about 40 mg/kg, about 50 mg/kg, about 60 mg/kg, about 70 mg/kg, about 80 mg/kg, about 90 mg/kg, about 100 mg/kg, about 125 mg/kg, about 150 mg/kg, about 175 mg/kg, or about 200 mg/kg per patient. 
     
     
         32 . A method of inhibiting BMP signaling comprising contacting cells with an antibody, or antigen-binding fragment thereof, wherein the antibody, or antigen-binding fragment thereof, comprises a heavy chain variable region having an amino acid sequence set forth as SEQ ID NO: 89 and a light chain variable region having an amino acid sequence set forth as SEQ ID NO: 93. 
     
     
         33 . A method of inhibiting BMP signaling in fibroblasts comprising contacting cells with an antibody, or antigen-binding fragment thereof, wherein the antibody, or antigen-binding fragment thereof, comprises a heavy chain variable region having an amino acid sequence set forth as SEQ ID NO: 89 and a light chain variable region having an amino acid sequence set forth as SEQ ID NO: 93, wherein:
 said heavy chain variable region further comprises one or more modifications selected from the group consisting of a substitution of glycine (G) by alanine (A) or serine (S) at position 49; a substitution of alanine (A) by isoleucine (I) at position 51; a substitution of lysine (K) by arginine (R) or asparagine (Q) at position 52b; a substitution of leucine (L) by valine (V) at position 78 utilizing the Kabat numbering system; and   the light chain variable region further comprises one or more modifications selected from the group consisting of a substitution of methionine (M) by leucine (L) at position 4; a substitution of alanine (A) by valine (V) at position 19; a substitution of threonine (T) by serine (S) at position 22; a substitution of alanine (A) by isoleucine (I) at position 48; and a substitution of threonine (T) by serine (S) at position 51 utilizing the Kabat numbering system,   wherein BMP signaling is inhibited.   
     
     
         34 . The method of  claim 33 , wherein the antibody, or antigen-binding fragment thereof, comprises a heavy chain variable region having an amino acid sequence set forth as SEQ ID NO: 88, 89, 90, 91 or 92; and a light chain variable region having an amino acid sequence set forth as SEQ ID NO: 93, 94, 95, 96, 97, 100, 102, or 103. 
     
     
         35 . A method of inhibiting SMAD 1/5/8 phosphorylation in fibroblasts comprising contacting cells with an antibody, or antigen-binding fragment thereof, wherein the antibody, or antigen-binding fragment thereof, comprises a heavy chain variable region having an amino acid sequence set forth as SEQ ID NO: 89 and a light chain variable region having an amino acid sequence set forth as SEQ ID NO: 93, wherein SMAD 1/5/8 phosphorylation is inhibited. 
     
     
         36 . A method of inhibiting SMAD 1/5/8 phosphorylation in fibroblasts comprising contacting cells with an antibody, or antigen-binding fragment thereof, wherein the antibody, or antigen-binding fragment thereof, comprises a heavy chain variable region having an amino acid sequence set forth as SEQ ID NO: 89 and a light chain variable region having an amino acid sequence set forth as SEQ ID NO: 93, wherein:
 said heavy chain variable region further comprises one or more modifications selected from the group consisting of a substitution of glycine (G) by alanine (A) or serine (S) at position 49; a substitution of alanine (A) by isoleucine (I) at position 51; a substitution of lysine (K) by arginine (R) or asparagine (Q) at position 52b; a substitution of leucine (L) by valine (V) at position 78 utilizing the Kabat numbering system; and   the light chain variable region further comprises one or more modifications selected from the group consisting of a substitution of methionine (M) by leucine (L) at position 4; a substitution of alanine (A) by valine (V) at position 19; a substitution of threonine (T) by serine (S) at position 22; a substitution of alanine (A) by isoleucine (I) at position 48; and a substitution of threonine (T) by serine (S) at position 51 utilizing the Kabat numbering system, wherein SMAD 1/5/8 phosphorylation is inhibited.   
     
     
         37 . The method of  claim 36 , wherein the antibody, or antigen-binding fragment thereof, comprises a heavy chain variable region having an amino acid sequence set forth as SEQ ID NO: 88, 89, 90, 91 or 92; and a light chain variable region having an amino acid sequence set forth as SEQ ID NO: 93, 94, 95, 96, 97, 100, 102, or 103. 
     
     
         38 . The method of any one of  claims 32 - 37 , wherein the antibody, or antigen-binding fragment thereof, blocks binding of BMP9 to endoglin.