Anti-wall teichoic antibodies and conjugates

Abstract

The invention provides anti-wall teichoic acid antibodies, antibiotic-linker intermediates, and antibody-antibiotic conjugate compound, and methods of making and using the same.

We claim: 
     
         1 . An antibiotic-linker intermediate selected from:
   X-L-abx   wherein:   abx is an antibiotic moiety selected from clindamycin, novobiocin, retapamulin, daptomycin, GSK-2140944, CG-400549, sitafloxacin, teicoplanin, triclosan, napthyridone, radezolid, doxorubicin, ampicillin, vancomycin, imipenem, doripenem, gemcitabine, dalbavancin, and azithromycin;   L is a peptide linker covalently attached to abx and X, and having the formula:
   -Str-Pep-Y- 
   
       where Str is a stretcher unit selected from the group consisting of C 1 -C 10  alkylene-, —C 3 -C 8  carbocyclo, —O—(C 1 -C 8  alkyl)-, -arylene-, alkylene-arylene-, -arylene-C 1 -C 10  alkylene-, —C 1 -C 10  alkylene-(C 3 -C 8  carbocyclo)-, —(C 3 -C 8  carbocyclo)-C 1 -C 10  alkylene-, —C 3 -C 8  heterocyclo-, —C 1 -C 10  alkylene-(C 3 -C 8  heterocyclo)-, —(C 3 -C 8  heterocyclo)-C 1 -C 10  alkylene-, —(CH 2 CH 2 O) r —, and —(CH 2 CH 2 O) r —CH 2 —; and r is an integer ranging from 1 to 10;
 Pep is a peptide of two to twelve amino acid residues, 
 Y is a spacer unit; and 
 X is a reactive functional group selected from maleimide, thiol, amino, bromide, bromoacetamido, iodoacetamido, p-toluenesulfonate, iodide, hydroxyl, carboxyl, pyridyl disulfide, and N-hydroxysuccinimide. 
 
     
     
         2 . The antibiotic-linker intermediate of  claim 1  wherein X is 
       
         
           
           
               
               
           
         
       
     
     
         3 . The antibiotic-linker intermediate of  claim 1  wherein Str is —(CH 2 ) 5 —. 
     
     
         4 . The antibiotic-linker intermediate of  claim 1  wherein Pep comprises two to twelve amino acid residues independently selected from glycine, alanine, phenylalanine, lysine, arginine, valine, and citrulline. 
     
     
         5 . The antibiotic-linker intermediate of  claim 4  wherein Pep is valine-citrulline. 
     
     
         6 . The antibiotic-linker intermediate of  claim 1  wherein Y comprises para-aminobenzyl or para-aminobenzyloxycarbonyl. 
     
     
         7 . The antibiotic-linker intermediate of  claim 1  wherein L is the peptide linker having the formula: 
       
         
           
           
               
               
           
         
         where AA1 and AA2 are independently selected from an amino acid side chain. 
       
     
     
         8 . The antibiotic-linker intermediate of  claim 7  wherein the amino acid side chain is independently selected from H, —CH 3 , —CH 2 (C 6 H 5 ), —CH 2 CH 2 CH 2 CH 2 NH 2 , —CH 2 CH 2 CH 2 NHC(NH)NH 2 , —CHCH(CH 3 )CH 3 , and —CH 2 CH 2 CH 2 NHC(O)NH 2 . 
     
     
         9 . The antibiotic-linker intermediate of  claim 7  wherein L is the peptide linker having the formula: 
       
         
           
           
               
               
           
         
       
     
     
         10 . The antibiotic-linker intermediate of  claim 1  wherein L is the peptide linker having the formula: 
       
         
           
           
               
               
           
         
       
     
     
         11 . The antibiotic-linker intermediate of  claim 10  wherein L is the peptide linker having the formula: 
       
         
           
           
               
               
           
         
       
     
     
         12 . The antibiotic-linker intermediate of  claim 11  wherein L is the peptide linker having the formula: 
       
         
           
           
               
               
           
         
       
     
     
         13 . The antibiotic-linker intermediate of  claim 12  wherein L is the peptide linker having the formula: 
       
         
           
           
               
               
           
         
       
     
     
         14 . The antibiotic-linker intermediate of  claim 10  wherein L is the peptide linker having the formula: 
       
         
           
           
               
               
           
         
       
     
     
         15 . The antibiotic-linker intermediate of  claim 14  wherein L is the peptide linker having the formula: 
       
         
           
           
               
               
           
         
       
     
     
         16 . The antibiotic-linker intermediate of  claim 10  wherein L is the peptide linker having the formula: 
       
         
           
           
               
               
           
         
         where R 7  is independently selected from H and C 1 -C 12  alkyl. 
       
     
     
         17 . The antibiotic-linker intermediate of  claim 1  selected from: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
     
     
         18 . A process for making an antibody-antibiotic conjugate compound comprising conjugating an anti-wall teichoic acid (WTA) monoclonal antibody to an antibiotic-linker intermediate,
 wherein the anti-wall teichoic acid (WTA) monoclonal antibody binds to  Staphylococcus aureus , and comprises: CDR L1 comprising the sequence of KSSQSIFRTSRNKNLLN (SEQ ID NO:99), CDR L2 comprising the sequence of WASTRKS (SEQ ID NO:100), and CDR L3 comprising the sequence of QQYFSPPYT (SEQ ID NO:101); and the VH of the anti-WTA monoclonal antibody comprises CDR H1 comprising the sequence of SFWMH (SEQ ID NO:102), CDR H2 comprising the sequence of FTNNEGTTTAYADSVRG (SEQ ID NO:103), and CDR H3 comprising the sequence of GEGGLDD (SEQ ID NO:118) or GDGGLDD (SEQ ID NO:104); and   the antibiotic-linker intermediate has the formula:
   X-L-abx 
   wherein:   abx is an antibiotic moiety selected from clindamycin, novobiocin, retapamulin, daptomycin, GSK-2140944, CG-400549, sitafloxacin, teicoplanin, triclosan, napthyridone, radezolid, doxorubicin, ampicillin, vancomycin, imipenem, doripenem, gemcitabine, dalbavancin, and azithromycin;   L is a peptide linker covalently attached to abx and X, and having the formula:
   -Str-Pep-Y- 
   where Str is a stretcher unit selected from the group consisting of C 1 -C 10  alkylene-, —C 3 -C 8  carbocyclo, —O—(C 1 -C 8  alkyl)-, -arylene-, alkylene-arylene-, -arylene-C 1 -C 10  alkylene-, —C 1 -C 10  alkylene-(C 3 -C 8  carbocyclo)-, —(C 3 -C 8  carbocyclo)-C 1 -C 10  alkylene-, —C 3 -C 8  heterocyclo-, —C 1 -C 10  alkylene-(C 3 -C 8  heterocyclo)-, —(C 3 -C 8  heterocyclo)-C 1 -C 10  alkylene-, —(CH 2 CH 2 O) r —, and —(CH 2 CH 2 O) r —CH 2 —; and r is an integer ranging from 1 to 10;   Pep is a peptide of two to twelve amino acid residues,   Y is a spacer unit; and   X is a reactive functional group selected from maleimide, thiol, amino, bromide, bromoacetamido, iodoacetamido, p-toluenesulfonate, iodide, hydroxyl, carboxyl, pyridyl disulfide, and N-hydroxysuccinimide.   
     
     
         19 . The process of  claim 18  wherein the anti-wall teichoic acid (WTA) monoclonal antibody is a cysteine engineered antibody. 
     
     
         20 . The process of  claim 19  further comprising the step of reducing the anti-wall teichoic acid (WTA) monoclonal antibody with DTT (dithiothreitol) or tricarbonylethylphosphine (TCEP) before conjugating. 
     
     
         21 . The process of  claim 18 , wherein the anti-wall teichoic acid (WTA) monoclonal antibody comprises a VL comprising the amino acid sequence SEQ ID NO:119. 
     
     
         22 . The process of  claim 18 , wherein the anti-wall teichoic acid (WTA) monoclonal antibody comprises a VH comprising the amino acid sequence of SEQ ID NO:156. 
     
     
         23 . The process of  claim 18 , wherein the anti-wall teichoic acid (WTA) monoclonal antibody comprises a VL and a VH, wherein the VL comprises the sequence of SEQ ID NO:119 and the VH comprises the sequence SEQ ID NO:156. 
     
     
         24 . The process of  claim 18 , wherein the anti-wall teichoic acid (WTA) monoclonal antibody comprises a light chain and a heavy chain sequence of
 (a) SEQ ID NO:123 and SEQ ID NO:147, or   (b) SEQ ID NO:145 and SEQ ID NO:147.   
     
     
         25 . The process of  claim 18  wherein X is 
       
         
           
           
               
               
           
         
       
     
     
         26 . The process of  claim 18  wherein the antibiotic-linker intermediate is selected from an antibiotic-linker intermediate of  claim 17 .