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 . An anti-TM4SF1 binding protein comprising:
 a) a heavy chain sequence comprising one or more amino acid substitutions in the sequence as set forth in the amino acid sequence of SEQ ID NO: 132, wherein the one or more amino acid substitutions are at positions that are selected from the group consisting of positions 1, 44, and 80 of SEQ ID NO: 132;   b) a light chain sequence comprising one or more amino acid substitutions in the sequence as set forth in the amino acid sequence of SEQ ID NO: 133, wherein the one or more amino acid substitutions are at amino acid positions 3, 26, and 62 of SEQ ID NO: 133; and wherein the amino acid at position 90 is tryptophan (W).   
     
     
         129 . The anti TM4SF1 binding protein of  claim 128 , wherein the one or more the amino acid substitutions of the heavy chain are selected from the group consisting of Q1E, D44G, and F80Y. 
     
     
         130 . The anti TM4SF1 binding protein of  claim 128 , wherein the one or more the amino acid substitutions of the light chain are selected from the group consisting of I3V, N26Q, and G62S. 
     
     
         131 . The anti-TM4SF1 binding protein of  claim 128 , wherein the heavy chain comprises the amino acid substitutions of Q1E, D44G, and F80Y, and wherein the light chain comprises the amino acid substitution of I3V, N26Q, and G62S. 
     
     
         132 . The anti-TM4SF1 binding protein of  claim 128 , wherein the heavy chain comprises the amino acid substitutions of Q1E, D44G, and F80Y, and wherein the light chain comprises the amino acid substitution of I3V and G62S. 
     
     
         133 . The anti-TM4SF1 binding protein of  claim 128 , comprising a heavy chain comprising an amino acid sequence that has at least 95% identity to a sequence selected from the group consisting of: SEQ ID NOs: 90, 92, 130, and 132; and a light chain comprising an amino acid sequence that has at least 95% identity to a sequence selected from the group consisting of: SEQ ID NO: 97, 99, 101, 103, 105, 131, and 133. 
     
     
         134 . The anti-TM4SF1 binding protein of  claim 128 , wherein the binding protein comprises an Fc region comprising a mutation at position N297. 
     
     
         135 . The anti-TM4SF1 binding protein of  claim 128 , comprising 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. 
     
     
         136 . The anti-TM4SF1 binding protein 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. 
     
     
         137 . The anti-TM4SF1 binding protein 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. 
     
     
         138 . The anti-TM4SF1 binding protein of  claim 128 , wherein the protein binds to human TM4SF1 with a K D  of about 1×10 −9  M or less in a standard flow cytometry assay using HUVEC cells. 
     
     
         139 . A antibody drug conjugate comprising: i) an antigen binding protein comprising the anti-TM4SF1 binding protein of  claim 128 ; and ii) a therapeutic molecule. 
     
     
         140 . The antibody drug conjugate of  claim 139 , wherein the therapeutic molecule is selected from a group consisting of a cytotoxic agent, a chemotherapeutic agent, a protein, a peptide, an antibody, a growth inhibitory agent, an anti-hormonal agent, or a combination thereof. 
     
     
         141 . The antibody drug conjugate of  claim 139 , wherein the therapeutic molecule is maytansine.