Tmem219 antibodies and therapeutic uses thereof

Abstract

The present invention relates to antibodies or antigen binding fragments thereof that bind specifically to the IGFBP3 receptor, namely TMEM219, to methods for their production, pharmaceutical compositions containing said antibodies, and uses thereof.

1 . An isolated antibody or antigen binding fragment thereof that binds to human TMEM219 receptor and inhibits or reduces the binding of IGFBP3 to said TMEM219 receptor. 
     
     
         2 . The isolated antibody or antigen binding fragment thereof according to  claim 1  that inhibits, reduces, or neutralizes the activation of the TMEM219 receptor induced by binding of IGFBP3. 
     
     
         3 . The isolated antibody or antigen binding fragment thereof according to  claim 1  that does not activate TMEM219 pathway upon binding to human TMEM219. 
     
     
         4 . The isolated antibody or antigen binding fragment thereof according to  claim 1  that is effective in preserving beta cells in diabetic subject and/or in preventing islet destruction, and/or in controlling blood glucose levels in an in vivo model. 
     
     
         5 . The isolated antibody or antigen binding fragment thereof according to  claim 1  that is effective in reducing acute and chronic colitis in an in vivo model. 
     
     
         6 . The isolated antibody or antigen binding fragment thereof according to  claim 1  that has at least one activity selected from:
 (a) increase in IGFBP3 treated healthy subject minigut growth; 
 (b) increase in IBD-patient minigut growth; 
 (c) increase in diabetic enteropathy serum treated healthy subject minigut growth; 
 (d) increase in expression of EphB2 and/or LGR5 in IGFBP3 treated healthy subject minigut; 
 (e) decrease in Caspase 8 expression in IGFBP3 treated healthy subject minigut; 
 (f) decrease in β-cell loss in IGFBP3 treated β-cell; 
 (g) increase in expression of insulin in IGFBP3 treated β-cell; 
 (h) inhibits or decreases DSS-induced intestinal cells apoptosis; 
 (i) restores expression of PCNA in DSS-treated colon; 
 (j) decrease in apoptosis of β-cell in IGFBP3 treated β-cell; 
 (k) decrease in insulitis score in an animal model of diabetes; 
 (l) decrease in diabetes onset in an animal model of diabetes; 
 (m) protects beta cell injury in an animal model of diabetes; and 
 (n) prevents beta cell loss in an animal model of diabetes. 
 
     
     
         7 . The isolated antibody or antigen binding fragment thereof according to  claim 6  wherein the increase in (a), (b) and (c) is by at least 20%; the increase in (d) and (e) is by at least 30%, optionally wherein by at least 50%; the decrease in (f) and the increase in (g) is by at least 10%. 
     
     
         8 . The isolated antibody or antigen binding fragment thereof according to  claim 1  comprising:
 (a) a heavy chain variable domain (VH) comprising:
 (i) a CDR1 sequence of the amino acid sequence selected from the group consisting of: SEQ ID NO:4, 1, 8, 10, 56, 59, 62, 65 or 68; 
 (ii) a CDR2 sequence of the amino acid sequence selected from the group consisting of: SEQ ID NO:5, 2, 11, 57, 60, 63, 66 or 69; and 
 (iii) a CDR3 sequence of the amino acid sequence selected from the group consisting of: SEQ ID NO:6, 3, 7, 9, 12, 13, 58, 61, 64, 67 or 70; and/or 
 
 (b) a light chain variable domain (VL) comprising:
 (i) a CDR1 sequence of the amino acid sequence selected from the group consisting of: SEQ ID NO:17, 14, 20, 23, 26, 29, 71, 77, 80, 82 or 85; 
 (ii) a CDR2 sequence of the amino acid sequence selected from the group consisting of: SEQ ID NO:18, 15, 21, 24, 27, 30, 72, 78, 83 or 86; and 
 (iii) a CDR3 sequence of the amino acid sequence selected from the group consisting of: SEQ ID NO:19, 16, 22, 25, 28, 31, 73, 74, 75, 76, 79, 81, 84, 87, 166 or 167. 
 
 
     
     
         9 . The isolated antibody or antigen binding fragment thereof according to  claim 1  comprising:
 (a) SEQ ID NO: 4 and SEQ ID NO: 5 and SEQ ID NO: 6 and SEQ ID NO: 17 and SEQ ID NO: 18 and SEQ ID NO: 19 or Kabat, IMGT, Chothia, AbM, or Contact CDRs of TC01 or of TC05; or 
 (b) SEQ ID NO: 4 and SEQ ID NO: 5 and SEQ ID NO: 6 and SEQ ID NO: 17 and SEQ ID NO: 18 and SEQ ID NO: 166 or Kabat, IMGT, Chothia, AbM, or Contact CDRs of TC03; or 
 (c) SEQ ID NO: 4 and SEQ ID NO: 5 and SEQ ID NO: 6 and SEQ ID NO: 17 and SEQ ID NO: 18 and SEQ ID NO: 167 or Kabat, IMGT, Chothia, AbM, or Contact CDRs of TC04; or 
 (d) SEQ ID NO: 68 and SEQ ID NO: 69 and SEQ ID NO: 70 and SEQ ID NO: 85 and SEQ ID NO: 86 and SEQ ID NO: 87 or Kabat, IMGT, Chothia, AbM, or Contact CDRs of TM1. 
 
     
     
         10 . The isolated antibody or antigen binding fragment thereof according to  claim 1  comprising:
 (a) a heavy chain variable domain (VH) comprising:
 (i) a CDR1 sequence of the amino acid sequence selected from the group consisting of a sequence as defined using abysis tool analysis; 
 (ii) a CDR2 sequence of the amino acid sequence selected from the group consisting of a sequence as defined using abysis tool analysis; and 
 (iii) a CDR3 sequence of the amino acid sequence selected from the group consisting of a sequence as defined using abysis tool analysis; and/or 
 
 (b) a light chain variable domain (VL) comprising:
 (i) a CDR1 sequence of the amino acid sequence selected from the group consisting of a sequence as defined using abysis tool analysis; 
 (ii) a CDR2 sequence of the amino acid sequence selected from the group consisting of a sequence as defined using abysis tool analysis; and 
 (iii) a CDR3 sequence of the amino acid sequence selected from the group consisting of a sequence as defined using abysis tool analysis. 
 
 
     
     
         11 . The isolated antibody or antigen binding fragment thereof according to  claim 1  comprising:
 (a) a heavy chain variable domain sequence of the amino acid sequence selected from the group consisting of: SEQ ID NO:32 to SEQ ID NO:37 or of SEQ ID NO:88 to SEQ ID NO:95, or of SEQ ID NO:168, SEQ ID NO:169 and SEQ ID NO:170; or 
 (b) a light chain variable domain sequence of the amino acid sequence selected from the group consisting of: SEQ ID NO: 38 to SEQ ID NO:43; or of SEQ ID NO: 96 to SEQ ID NO: 103 or of SEQ ID NO: 171, SEQ ID NO: 172 or SEQ ID NO: 173; or 
 (c) the heavy chain variable domain of (a) and the light chain variable domain of (b). 
 
     
     
         12 . The isolated antibody or antigen binding fragment thereof according to  claim 1  comprising VH and VL regions together selected from the group consisting of antibody TC01, TC03, TC04, TC05, TA02, TC02, TD01, TE01, TG02, TM1, TE02.1, TE02.2, TE02.3, TE03, TE04, TE07, TE10, or an antigen binding fragment thereof. 
     
     
         13 . The isolated antibody or antigen binding fragment thereof according to  claim 10 , wherein the antibody or antigen binding fragment thereof comprises VH CDR1, CDR2 and CDR3 selected from Table 3.3 and VL CDR1, CDR2 and CDR3 selected from Table 3.4. 
     
     
         14 . The isolated antibody or antigen binding fragment thereof according to  claim 1  having an affinity constant lower than or equal to 10 −7  M for human TMEM. 
     
     
         15 . An isolated antibody or antigen binding fragment thereof that:
 (a) binds specifically to an epitope on TMEM, wherein the epitope is the same or similar epitope as the epitope recognized by the antibody TC01, TC03, TC04, TC05, TA02, TC02, TD01, TE01, TG02, TM1, TE02.1, TE02.2, TE02.3, TE03, TE04, TE07, TE10 as defined in Tables 2-19 and Tables 3.1-3.4; or   (b) cross-competes for binding with the monoclonal antibody TC01, TC03, TC04, TC05, TA02, TC02, TD01, TE01, TG02, TM1, TE02.1, TE02.2, TE02.3, TE03, TE04, TE07, TE10 as defined in Tables 2-19 and Tables 3.1-3.4; or   (c) shows the same or similar binding affinity or specificity, or both, as any of TC01, TC03, TC04, TC04, TA02, TC02, TD01, TE01, TG02, TM1, TE02.1, TE02.2, TE02.3, TE03, TE04, TE07, TE10 as defined in Tables 2-19 and Tables 3.1-3.4; or   (d) has one or more biological properties of an antibody molecule described herein, e.g., an antibody molecule chosen from, e.g., any of TC01, TC03, TC04, TC04, TA02, TC02, TD01, TE01, TG02, TM1, TE02.1, TE02.2, TE02.3, TE03, TE04, TE07, TE10 as defined in Tables 2-19 and Tables 3.1-3.4; or   (e) has one or more pharmacokinetic properties of an antibody molecule described herein, e.g., an antibody molecule chosen from, e.g., any of TC01, TC03, TC04, TC04, TA02, TC02, TD01, TE01, TG02, TM1, TE02.1, TE02.2, TE02.3, TE03, TE04, TE07, TE10 as defined in Tables 2-19 and Tables 3.1-3.4.   
     
     
         16 . The isolated antibody or antigen binding fragment thereof according to  claim 1  being a human or a humanized antibody. 
     
     
         17 . The isolated antibody or antigen binding fragment thereof according to  claim 1 , wherein the antibody or antigen binding fragment thereof is an IgG2 or IgG4 antibody. 
     
     
         18 . An isolated polynucleotide comprising at least one sequence that encodes the antibody or antigen binding fragment thereof according to  claim 1 . 
     
     
         19 . A vector comprising the polynucleotide according to  claim 18 , optionally wherein the vector is selected from the group consisting of a plasmid, a viral vector, a non-episomal mammalian vector, an expression vector and a recombinant expression vector. 
     
     
         20 . An isolated cell comprising the polynucleotide according to  claim 18 , optionally wherein the isolated cell is a hybridoma or a Chinese Hamster Ovary (CHO) cell or a Human Embryonic Kidney cells (HEK293). 
     
     
         21 . A method of treating a disorder, comprising: administering a therapeutically effective amount of the antibody or antigen binding fragment thereof according to  claim 1 , optionally wherein the disorder is selected from: diabetes, intestinal and/or bowel disorder, malabsorption syndrome, cachexia or diabetic enteropathy. 
     
     
         22 . A pharmaceutical composition comprising the isolated antibody or antigen binding fragment thereof according to  claim 1 , and a pharmaceutically acceptable carrier, optionally wherein for use in the treatment of: diabetes, intestinal and/or bowel disorder, malabsorption syndrome, cachexia or diabetic enteropathy. 
     
     
         23 . A method of inhibiting the binding of IGFBP3 to TMEM219 receptor, comprising contacting TMEM219 with the antibody according to  claim 1 . 
     
     
         24 . The isolated antibody or antigen binding fragment thereof according to  claim 17 , wherein the antibody or antigen binding fragment thereof is a human IgG2 or human IgG4. 
     
     
         25 . The isolated antibody or antigen binding fragment thereof according to  claim 17 , wherein the antibody or antigen binding fragment thereof is selected from the group consisting of an IgG2 kappa antibody, an IgG2 lambda antibody, an IgG4 kappa antibody or an IgG4 lambda antibody, or an antigen binding fragment thereof. 
     
     
         26 . The isolated polynucleotide according to  claim 18 , wherein said polynucleotide is a cDNA. 
     
     
         27 . The method of treating a disorder of  claim 21 , the diabetes disorder wherein it is Type I or Type II. 
     
     
         28 . The method of treating a disorder of  claim 21 , the intestinal and/or bowel disorder wherein it is inflammatory bowel disease, celiac disease, ulcerative colitis, Crohn's disease or intestinal obstruction. 
     
     
         29 . The vector according to  claim 19 , wherein the vector is a plasmid. 
     
     
         30 . The vector according to  claim 19 , wherein the vector is a viral vector. 
     
     
         31 . The vector according to  claim 19 , wherein the vector is a non-episomal mammalian vector. 
     
     
         32 . The vector according to  claim 19 , wherein the vector is an expression vector. 
     
     
         33 . The vector according to  claim 19 , wherein the vector is a recombinant expression vector. 
     
     
         34 . The isolated cell according to  claim 20 , wherein the isolated cell is hybridoma. 
     
     
         35 . The isolated cell according to  claim 20 , wherein the isolated cell is Chinese Hamster Ovary (CHO) cell. 
     
     
         36 . The isolated cell according to  claim 20 , wherein the isolated cell is Human Embryonic Kidney (HEK293) cell. 
     
     
         37 . A method of treating a disorder, comprising administering a therapeutically effective amount of a pharmaceutical composition comprising the isolated antibody or antigen binding fragment thereof according to  claim 1 , and a pharmaceutically acceptable carrier, optionally wherein for the use in the treatment of: diabetes, intestinal and/or bowel disorder, malabsorption syndrome, cachexia or diabetic enteropathy, optionally wherein the intestinal and/or bowel disorder is inflammatory bowel disease, celiac disease, ulcerative colitis, Crohn's disease or intestinal obstruction. 
     
     
         38 . The isolated antibody or antigen binding fragment thereof according to  claim 11 , wherein the heavy chain variable domain sequence comprises or has the amino acid sequence of SEQ ID NO:33 and the light chain variable domain sequence comprises or has the amino acid sequence of SEQ ID NO:39 
     
     
         39 . The isolated antibody or antigen binding fragment thereof according to  claim 1  comprising a light chain constant domain or an amino acid sequence with SEQ ID NO: 123; and a heavy chain constant domain of an amino acid sequence with SEQ ID NO: 120 wherein said heavy chain constant region is a human IgG4 including a Ser to Pro substitution at position 228 and a Leu to Glu substitution at position 235.