Anti-tm4sf1 antibodies and methods of using same

Abstract

Anti-TM4SF1 antibodies, and antigen-binding fragments thereof, are described that bind to an epitope on the ECL2 loop of human TM4SF1. Methods of use of said antibodies and fragments are also described, including for the inhibition of metastasis.

1 .- 127 . (canceled) 
     
     
         128 . A method of treating disease or disorder in a subject, wherein the disease or disorder is characterized by abnormal endothelial cell (EC)-cell interaction, said method comprising administering to the subject an anti-TM4SF1 binding protein comprising:
 a heavy chain variable domain comprising a CDR3 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 8, 20, 32, 44, 56, 68, 80, 96, 119, 120, or 121; a CDR2 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 7, 19, 31, 43, 55, 67, 79, 95, or 117; and a CDR1 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 6, 18, 30, 42, 54, 66, 78, or 94; and   a light chain variable domain comprising a CDR3 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 14, 26, 38, 50, 62, 74, 86, 110, or 111; a CDR2 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 13, 25, 37, 49, 61, 73, 85, or 109; and a CDR1 comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 12, 24, 36, 48, 60, 72, 84, 107, 108, 125, 126, or 127.   
     
     
         129 . The method of  claim 128 , wherein the anti-TM4SF1 binding protein comprises:
 the heavy chain variable domain comprises a CDR3 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 8, a CDR2 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 7, and a CDR1 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 6; and   wherein the light chain variable domain comprises a CDR3 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 14, a CDR2 domain comprising an amino acid that has at least 75% identity to SEQ ID NO: 13, and a CDR1 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 12.   
     
     
         130 . The method of  claim 128 , wherein the anti-TM4SF1 binding protein comprises:
 the heavy chain variable domain comprises a CDR3 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 20, a CDR2 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 19, and a CDR1 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 18; and   wherein the light chain variable domain comprises a CDR3 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 26, a CDR2 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 25, and a CDR1 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 24.   
     
     
         131 . The method of  claim 128 , wherein the anti-TM4SF1 binding protein comprises:
 the heavy chain variable domain comprises a CDR3 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 32, a CDR2 domain comprising an amino acid sequence an amino acid sequence that has at least 75% identity to SEQ ID NO: 31, and a CDR1 domain comprising an amino acid sequence an amino acid sequence that has at least 75% identity to SEQ ID NO: 30; and   wherein the light-chain variable domain comprises a CDR3 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 38, a CDR2 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 37, and a CDR1 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 36.   
     
     
         132 . The method of  claim 128 , wherein the anti-TM4SF1 binding protein comprises:
 the heavy chain variable domain comprises a CDR3 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 44, a CDR2 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 43, and a CDR1 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 42; and   the light chain variable domain comprises a CDR3 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 50, a CDR2 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 49, and a CDR1 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 48.   
     
     
         133 . The method of  claim 128 , wherein the anti-TM4SF1 binding protein comprises:
 the heavy chain variable domain comprises a CDR3 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 56, a CDR2 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 55, and a CDR1 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 54; and   wherein the light chain variable domain comprises a CDR3 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 62, a CDR2 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 61, and a CDR1 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 60.   
     
     
         134 . The method of  claim 128 , wherein the anti-TM4SF1 binding protein comprises:
 the heavy chain variable domain comprises a CDR3 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 68, a CDR2 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 67, and a CDR1 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 66; and   wherein the light chain variable domain comprising a CDR3 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 74, a CDR2 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 73, and a CDR1 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 72.   
     
     
         135 . The method of  claim 128 , wherein the anti-TM4SF1 binding protein comprises:
 the heavy chain variable domain comprises a CDR3 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 80, a CDR2 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 79, and a CDR1 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 78; and   wherein the light chain variable domain comprises a CDR3 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 86, a CDR2 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 85, and a CDR1 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 84.   
     
     
         136 . The anti-TM4SF1 binding protein of  claim 128 , wherein the heavy chain variable domain comprises the CDR3 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 96, the CDR2 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 95, and the CDR1 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 94; and wherein the light chain variable domain comprises the CDR3 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 110, the CDR2 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 109, and the CDR1 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 108. 
     
     
         137 . The anti-TM4SF1 binding protein of  claim 128 , wherein the heavy chain variable domain comprises the CDR3 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 96, the CDR2 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 95, and the CDR1 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 94; and wherein the light chain variable domain comprises the CDR3 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 110, the CDR2 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 109, and the CDR1 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 107. 
     
     
         138 . The anti-TM4SF1 binding protein of  claim 128 , wherein the heavy chain variable domain comprises the CDR3 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 96, the CDR2 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 95, and the CDR1 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 94; and wherein the light chain variable domain comprises the CDR3 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 111, the CDR2 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 109, and the CDR1 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 107. 
     
     
         139 . The anti-TM4SF1 binding protein of  claim 128 , wherein the heavy chain variable domain comprises the CDR3 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 96, the CDR2 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 95, and the CDR1 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 94; and wherein the light chain variable domain comprises the CDR3 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 111, the CDR2 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 109, and the CDR1 domain comprising an amino acid sequence that has at least 75% identity to SEQ ID NO: 108. 
     
     
         140 . The method of  claim 128 , wherein the anti-TM4SF1 binding protein comprises:
 a heavy chain comprising an amino acid sequence that has at least 75% identity to a sequence selected from the group consisting of: SEQ ID NOs: 3, 15, 27, 39, 51, 63, 75, 90, 92, 112, 114, 130, and 132, and   a light chain comprising an amino acid sequence that has at least 75% identity to a sequence selected from the group consisting of: SEQ ID NOs: 9, 21, 33, 45, 57, 69, 81, 97, 99, 101, 103, 105, 122, 131, and 133.   
     
     
         141 . The method of  claim 128 , wherein the anti-TM4SF1 binding protein comprises an Fc region comprising a mutation at position N297. 
     
     
         142 . The method of  claim 128 , wherein the anti-TM4SF1 binding protein comprises an antigen-binding fragment of an anti-TM4SF1 antibody, wherein the antigen-binding fragment comprises a Fab, a Fab′, a F(ab′)2, an Fv, or an scFv. 
     
     
         143 . The method of  claim 128 , wherein the binding of the protein to human TM4SF1 is not dependent on glycosylation of the ECL2 loop of human TM4SF1, wherein the human TM4SF1 comprises a sequence as set forth in SEQ ID NO: 134. 
     
     
         144 . The method of  claim 128 , wherein the protein binds to a cynomolgus TM4SF1 with a KD about 1×10 −8  M or less in a standard flow cytometry assay using HEK293 overexpressing cells. 
     
     
         145 . The method of  claim 128 , wherein the protein binds to human TM4SF1 with a KD of about 1×10 −9  M or less in a standard flow cytometry assay using HUVEC cells. 
     
     
         146 . The method of  claim 128 , wherein the EC-cell interaction comprises one or more of EC-mesenchymal stem cell, EC-fibroblast, EC-smooth muscle cell, EC-tumor cell, EC-leukocyte, EC-adipose cell, and EC-neuronal cell interactions. 
     
     
         147 . The method of  claim 128 , wherein the disease or disorder comprises an inflammatory disease or a cancer. 
     
     
         148 . The method of  claim 128 , wherein the administering comprises administering through an intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, pulmonary, or transmucosal route.