Anti-cd79b antibodies and immunoconjugates and methods of use

Abstract

The present invention is directed to compositions of matter useful for the treatment of hematopoietic tumor in mammals and to methods of using those compositions of matter for the same.

1 - 31 . (canceled) 
     
     
         32 . A method for making an antibody-drug conjugate compound comprising an anti-CD79b cysteine engineered antibody (Ab) comprising one or more engineered cysteines, and an auristatin or maytansinoid drug moiety (D), wherein the antibody is attached through the one or more engineered cysteine amino acids by a linker moiety (L) to D; the compound having Formula I:
   Ab-(L-D) p   I
   wherein the antibody comprises:   (I) at least one HVR sequence selected from the group consisting of:
 (i) HVR-L1 comprising sequence A1-A15, wherein A1-A15 is KASQSVDYDGDSFLN (SEQ ID NO: 131); 
 (ii) HVR-L2 comprising sequence B1-B7, wherein B1-B7 is AASNLES (SEQ ID NO: 132); 
 (iii) HVR-L3 comprising sequence C1-C9, wherein C1-C9 is QQSNEDPLT (SEQ ID NO: 133); 
 (iv) HVR—H1 comprising sequence D1-D10, wherein D1-D10 is GYTFSSYWIE (SEQ ID NO: 134); 
 (v) HVR—H2 comprising sequence E1-E18, wherein E1-E18 is GEILPGGGDTNYNEIFKG (SEQ ID NO: 135); and 
 (vi) HVR—H3 comprising sequence F1-F10, wherein F1-F10 is TRRVPVYFDY (SEQ ID NO: 136); and 
   (II) at least one variant HVR, wherein the variant HVR comprises modification of at least one residue of the sequence depicted in SEQ ID NO: 131, 132, 133, 134, 135 or 136;   wherein p is 1, 2, 3, or 4; the method comprising the steps of:   (a) reacting an engineered cysteine group of the antibody with a linker reagent to form antibody-linker intermediate Ab-L; and   (b) reacting Ab-L with an activated drug moiety D; whereby the antibody-drug conjugate is formed;   or comprising the steps of:   (c) reacting a nucleophilic group of a drug moiety with a linker reagent to form drug-linker intermediate D-L; and   (d) reacting D-L with an engineered cysteine group of the antibody; whereby the antibody-drug conjugate is formed.   
     
     
         33 . The method of  claim 32 , further comprising the step of expressing the antibody in Chinese hamster ovary (CHO) cells. 
     
     
         34 . The method of  claim 32 , further comprising the step of treating the expressed antibody with a reducing agent. 
     
     
         35 . The method of  claim 34 , wherein the reducing agent is selected from tricarbonylethylphosphine (TCEP) and dithiothreitol (DTT). 
     
     
         36 . The method of  claim 34 , further comprising the step of treating the expressed antibody with an oxidizing agent, after treating with the reducing agent. 
     
     
         37 . The method of  claim 36 , wherein the oxidizing agent is selected from copper sulfate, dehydroascorbic acid, and air. 
     
     
         38 . The method of  claim 32 , wherein the one or more engineered cysteines are at one or more positions selected from 15, 110, 114, 121, 127, 168, and 205 of the light chain according to Kabat numbering convention, and 5, 23, 84, and 112 of the heavy chain according to Kabat numbering convention, and 118, 120, 282, 375, and 400 of the heavy chain according to EU numbering convention.