Anti-notch3 antibodies and antibody-drug conjugates

Abstract

The present invention provides for anti-Notch3 antibodies, anti-Notch3 antibody-drug conjugates and methods for preparing and using the same.

1 .- 45 . (canceled) 
     
     
         46 . A method of treating a condition associated with Notch3 expression in a patient in need thereof, comprising administering to the patient an effective amount of a pharmaceutical composition comprising an antibody-drug conjugate of the formula Ab-(L-D)p, wherein:
 (a) Ab is an antibody, or antigen-binding fragment thereof, that binds to Notch3;   (b) L-D is a linker-drug moiety, wherein L is a linker and D is a drug, and wherein D is an auristatin; and   (c) p is an integer from 1 to 12.   
     
     
         47 . The method of  claim 46 , wherein Ab comprises:
 a VH CDR1, a VH CDR2, and a VH CDR3 of a heavy chain variable region (VH) comprising the amino acid sequence set forth in SEQ ID NO: 13; and a VL CDR1, a VL CDR2, and a VL CDR3 of a light chain variable region (VL) comprising the amino acid sequence set forth in SEQ ID NO: 25; or   a VH CDR1, a VH CDR2, and a VH CDR3 of a heavy chain variable region (VH) comprising the amino acid sequence set forth in SEQ ID NO: 37; and a VL CDR1, a VL CDR2, and a VL CDR3 of a light chain variable region (VL) comprising the amino acid sequence set forth in SEQ ID NO: 49.   
     
     
         48 . The method of  claim 46 , wherein Ab comprises:
 a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 15 or 16; a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 19 or 20; a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 23; a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 27; a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 29; and a VL CDR3 comprising the amino acid sequence of SEQ ID NO: 31; or   a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 39 or 40; a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 43 or 44; a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 47; a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 51; a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 53; and a VL CDR3 comprising the amino acid sequence of SEQ ID NO: 55.   
     
     
         49 . The method of  claim 48 , wherein Ab comprises:
 a heavy chain variable region having an amino acid sequence that is at least 90% identical to SEQ ID NO: 13 or a light chain variable region having an amino acid sequence that is at least 90% identical to SEQ ID NO: 25; or   a heavy chain variable region having an amino acid sequence that is at least 90% identical to SEQ ID NO: 37 or a light chain variable region having an amino acid sequence that is at least 90% identical to SEQ ID NO: 49.   
     
     
         50 . The method of  claim 49 , wherein Ab comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 13; or a heavy chain variable region amino acid sequence of SEQ ID NO: 37. 
     
     
         51 . The method of  claim 49 , wherein Ab comprises a light chain variable region amino acid sequence of SEQ ID NO: 25 or a light chain variable region amino acid sequence of SEQ ID NO: 49. 
     
     
         52 . The method of  claim 50 , wherein Ab comprises a heavy chain amino acid sequence of SEQ ID NO: 33; or a heavy chain amino acid sequence of SEQ ID NO: 57. 
     
     
         53 . The method of  claim 51 , wherein Ab comprises a light chain amino acid sequence of SEQ ID NO: 35; or a light chain amino acid sequence of SEQ ID NO: 59. 
     
     
         54 . The method of  claim 46 , wherein L is selected from the group consisting of maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (vc), maleimidocaproyl (mc), maleimido-heptanoyl (me) and maleimido-Peg6C2 (MalPeg6C2). 
     
     
         55 . The method of  claim 46 , wherein D is a drug selected from the group consisting of:
 (i) 0101 having the formula:   
       
         
           
           
               
               
           
         
         (ii) 6780 having the formula: 
       
       
         
           
           
               
               
           
         
         (iii) 0131 having the formula: 
       
       
         
           
           
               
               
           
         
         (iv) 3377 having the formula: 
       
       
         
           
           
               
               
           
         
       
       and
 (v) 8261 having the formula: 
 
       
         
           
           
               
               
           
         
       
     
     
         56 . The method of  claim 46 , wherein L-D is selected from the group consisting of:
 vc0101 having the formula:   
       
         
           
           
               
               
           
         
         and vc6780 having the formula: 
       
       
         
           
           
               
               
           
         
       
     
     
         57 . The method of  claim 46 , wherein the condition is cancer. 
     
     
         58 . The method of  claim 57 , wherein the cancer is a solid tumor cancer. 
     
     
         59 . The method of  claim 58 , wherein the solid tumor cancer is selected from the group consisting of lung cancer, breast cancer, ovarian cancer, stomach cancer, esophageal cancer, cervical cancer, head and neck cancer, bladder cancer, liver cancer, skin cancer and sarcoma. 
     
     
         60 . The method of  claim 57 , wherein the cancer is a blood cancer. 
     
     
         61 . The method of  claim 60  wherein the blood cancer is selected from the group consisting of T-cell malignancies, T-cell leukemia, T-cell lymphoma, T-cell acute lymphoblastic leukemia, multiple myeloma, B-cell malignancies, myeloid malignancies, acute myeloid leukemia and chronic myeloid leukemia. 
     
     
         62 . A method of inhibiting tumor growth or progression in a patient having a Notch3 expressing cancer, comprising administering to the patient an effective amount of a pharmaceutical composition comprising an antibody-drug conjugate of the formula Ab-(L-D)p, wherein:
 (a) Ab is an antibody, or antigen-binding fragment thereof, that binds to Notch3;   (b) L-D is a linker-drug moiety, wherein L is a linker and D is a drug, and wherein D is an auristatin; and   (c) p is an integer from 1 to 12.   
     
     
         63 . The method of  claim 62 , wherein Ab comprises:
 a VH CDR1, a VH CDR2, and a VH CDR3 of a heavy chain variable region (VH) comprising the amino acid sequence set forth in SEQ ID NO: 13; and a VL CDR1, a VL CDR2, and a VL CDR3 of a light chain variable region (VL) comprising the amino acid sequence set forth in SEQ ID NO: 25; or   a VH CDR1, a VH CDR2, and a VH CDR3 of a heavy chain variable region (VH) comprising the amino acid sequence set forth in SEQ ID NO: 37; and a VL CDR1, a VL CDR2, and a VL CDR3 of a light chain variable region (VL) comprising the amino acid sequence set forth in SEQ ID NO: 49.   
     
     
         64 . The method of  claim 62 , wherein Ab comprises:
 a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 15 or 16; a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 19 or 20; a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 23; a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 27; a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 29; and a VL CDR3 comprising the amino acid sequence of SEQ ID NO: 31; or   a VH CDR1 comprising the amino acid sequence of SEQ ID NO: 39 or 40; a VH CDR2 comprising the amino acid sequence of SEQ ID NO: 43 or 44; a VH CDR3 comprising the amino acid sequence of SEQ ID NO: 47; a VL CDR1 comprising the amino acid sequence of SEQ ID NO: 51; a VL CDR2 comprising the amino acid sequence of SEQ ID NO: 53; and a VL CDR3 comprising the amino acid sequence of SEQ ID NO: 55.   
     
     
         65 . The method of  claim 64 , wherein Ab comprises:
 a heavy chain variable region having an amino acid sequence that is at least 90% identical to SEQ ID NO: 13 or a light chain variable region having an amino acid sequence that is at least 90% identical to SEQ ID NO: 25; or   a heavy chain variable region having an amino acid sequence that is at least 90% identical to SEQ ID NO: 37 or a light chain variable region having an amino acid sequence that is at least 90% identical to SEQ ID NO: 49.   
     
     
         66 . The method of  claim 65 , wherein Ab comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 13; or a heavy chain variable region amino acid sequence of SEQ ID NO: 37. 
     
     
         67 . The method of  claim 65 , wherein Ab comprises a light chain variable region amino acid sequence of SEQ ID NO: 25 or a light chain variable region amino acid sequence of SEQ ID NO: 49. 
     
     
         68 . The method of  claim 66 , wherein Ab comprises a heavy chain amino acid sequence of SEQ ID NO: 33; or a heavy chain amino acid sequence of SEQ ID NO: 57. 
     
     
         69 . The method of  claim 67 , wherein Ab comprises a light chain amino acid sequence of SEQ ID NO: 35; or a light chain amino acid sequence of SEQ ID NO: 59. 
     
     
         70 . The method of  claim 62 , wherein L is selected from the group consisting of maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (vc), maleimidocaproyl (mc), maleimido-heptanoyl (me) and maleimido-Peg6C2 (MalPeg6C2). 
     
     
         71 . The method of  claim 62 , wherein D is a drug selected from the group consisting of:
 (i) 0101 having the formula:   
       
         
           
           
               
               
           
         
         (ii) 6780 having the formula: 
       
       
         
           
           
               
               
           
         
         (iii) 0131 having the formula: 
       
       
         
           
           
               
               
           
         
         (iv) 3377 having the formula: 
       
       
         
           
           
               
               
           
         
       
       and
 (v) 8261 having the formula: 
 
       
         
           
           
               
               
           
         
       
     
     
         72 . The method of  claim 62 , wherein L-D is selected from the group consisting of:
 vc0101 having the formula:   
       
         
           
           
               
               
           
         
         and vc6780 having the formula: 
       
       
         
           
           
               
               
           
         
       
     
     
         73 . The method of  claim 62 , wherein the cancer is a solid tumor cancer. 
     
     
         74 . The method of  claim 73 , wherein the solid tumor cancer is selected from the group consisting of lung cancer, breast cancer, ovarian cancer, stomach cancer, esophageal cancer, cervical cancer, head and neck cancer, bladder cancer, liver cancer, skin cancer and sarcoma. 
     
     
         75 . The method of  claim 62 , wherein the cancer is a blood cancer. 
     
     
         76 . The method of  claim 75  wherein the blood cancer is selected from the group consisting of T-cell malignancies, T-cell leukemia, T-cell lymphoma, T-cell acute lymphoblastic leukemia, multiple myeloma, B-cell malignancies, myeloid malignancies, acute myeloid leukemia and chronic myeloid leukemia.