Anti-b7-h4 antibodies and immunoconjugates

Abstract

The invention provides anti-B7-H4 antibodies and immunoconjugates and methods of using the same.

1 - 19 . (canceled) 
     
     
         20 . An immunoconjugate comprising an antibody that binds to B7-H4 and a cytotoxic agent, wherein the antibody comprises:
 (a)(i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 29, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 30, and (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 31; (iv) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 32, (v) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 33, and (vi) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 34; or   (b)(i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 58, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 59, and (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 60; (iv) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 61, (v) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 62, and (vi) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 63.   
     
     
         21 . The immunoconjugate of  claim 20  having the formula Ab-(L-D)p, wherein:
 (a) Ab is the antibody; 
 (b) L is a linker; 
 (c) D is a cytotoxic agent selected from a maytansinoid, an auristatin, a calicheamicin, a pyrrolobenzodiazepine, and a nemorubicin derivative; and 
 (d) p ranges from 1-8. 
 
     
     
         22 . The immunoconjugate of  claim 20 , wherein the cytotoxic agent is an auristatin. 
     
     
         23 . The immunoconjugate of  claim 21 , wherein D has formula D E    
       
         
           
           
               
               
           
         
         and wherein R 2  and R 6  are each methyl, R 3  and R 4  are each isopropyl, R 5  is H, R 7  is sec-butyl, each R 8  is independently selected from CH 3 , O—CH 3 , OH, and H; R 9  is H; and R 18  is —C(R 8 ) 2 —C(R 8 ) 2 -aryl. 
       
     
     
         24 . The immunoconjugate of  claim 20 , wherein the cytotoxic agent is MMAE. 
     
     
         25 . The immunoconjugate of  claim 21 , wherein D is a pyrrolobenzodiazepine of Formula A: 
       
         
           
           
               
               
           
         
         wherein the dotted lines indicate the optional presence of a double bond between C1 and C2 or C2 and C3; 
         R 2  is independently selected from H, OH, ═O, ═CH 2 , CN, R, OR, ═CH—R D , ═C(R D ) 2 , O—SO 2 —R, CO 2 R and COR, and optionally further selected from halo or dihalo, wherein R D  is independently selected from R, CO 2 R, COR, CHO, CO 2 H, and halo; 
         R 6  and R 9  are independently selected from H, R, OH, OR, SH, SR, NH 2 , NHR, NRR′, NO 2 , Me 3 Sn and halo; 
         R 7  is independently selected from H, R, OH, OR, SH, SR, NH 2 , NHR, NRR′, NO 2 , Me 3 Sn and halo; 
         Q is independently selected from O, S and NH, and 
         R 11  is either H or R; or Q is O and R 11  is SO 3 M, where M is a metal cation; 
         R and R′ are each independently selected from optionally substituted C 1-8  alkyl, C 3-8  heterocyclyl and C 5-20  aryl groups, and optionally in relation to the group NRR′, R and R′ together with the nitrogen atom to which they are attached form an optionally substituted 4-, 5-, 6- or 7-membered heterocyclic ring; 
         R 12 , R 16 , R 19  and R 17  are as defined for R 2 , R 6 , R 9  and R 7  respectively; 
         R″ is a C 3-12  alkylene group, which chain may be interrupted by one or more heteroatoms and/or aromatic rings that are optionally substituted; and 
         X and X′ are independently selected from O, S and N(H). 
       
     
     
         26 . The immunoconjugate of  claim 25 , wherein D has the structure: 
       
         
           
           
               
               
           
         
         wherein n is 0 or 1. 
       
     
     
         27 . The immunoconjugate of  claim 20 , wherein the cytotoxic agent is a nemorubicin derivative. 
     
     
         28 . The immunoconjugate of  claim 21 , wherein D has a structure selected from: 
       
         
           
           
               
               
           
         
       
     
     
         29 . The immunoconjugate of  claim 21 , wherein the linker is cleavable by a protease. 
     
     
         30 . The immunoconjugate of  claim 29 , wherein the linker comprises a valine-citrulline dipeptide, an alanine-phenylalanine dipeptide, a phenylalanine-lysine dipeptide, a phenylalanine-homolysine dipeptide, or a N-methyl-valine-citrulline dipeptide. 
     
     
         31 . The immunoconjugate of  claim 21 , wherein the linker is acid-labile. 
     
     
         32 . The immunoconjugate of  claim 31 , wherein the linker comprises hydrazone. 
     
     
         33 . The immunoconjugate of  claim 21  having the formula: 
       
         
           
           
               
               
           
         
         wherein S is a sulfur atom. 
       
     
     
         34 . The immunoconjugate of  claim 21  having the formula: 
       
         
           
           
               
               
           
         
       
     
     
         35 . The immunoconjugate of  claim 21  having a formula selected from: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
       
     
     
         36 . The immunoconjugate of  claim 21 , wherein p ranges from 2-5. 
     
     
         37 . The immunoconjugate of  claim 20 , wherein the antibody comprises:
 (a) a VH sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 28; or   (b) a VL sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 27; or   (c) a VH sequence as in (a) and a VL sequence as in (b); or   (d) a VH sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 56; or   (e) a VL sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 55; or   (f) a VH sequence as in (d) and a VL sequence as in (e); or   (g) a VL sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 57; or   (h) a VH sequence as in (d) and a VL sequence as in (g).   
     
     
         38 . The immunoconjugate of  claim 20 , wherein comprising the antibody of claim  15  comprises (a) a VH sequence of SEQ ID NO: 28 and a VL sequence of SEQ ID NO: 27; or (b) a VH sequence of SEQ ID NO: 56 and a VL sequence of SEQ ID NO: 55; or (c) a VH sequence of SEQ ID NO: 56 and a VL sequence of SEQ ID NO: 57. 
     
     
         39 . A pharmaceutical formulation comprising the immunoconjugate of  claim 20  and a pharmaceutically acceptable carrier. 
     
     
         40 . The pharmaceutical formulation of  claim 39 , further comprising an additional therapeutic agent. 
     
     
         41 . The pharmaceutical formulation of  claim 40 , wherein the additional therapeutic agent is Avastin® (bevacizumab). 
     
     
         42 .- 57 . (canceled)