Humanized or chimeric cd3 antibodies

Abstract

The present invention relates to humanized or chimeric antibodies binding CD3. It furthermore relates to bispecific antibodies, compositions, pharmaceutical compositions, use of said antibodies in the treatment of a disease, and method of treatment.

1 - 66 . (canceled) 
     
     
         67 . A nucleic acid comprising a nucleotide sequence encoding (a) a heavy chain and/or (b) a light chain of a humanized or chimeric antibody which binds to human CD3, wherein the antibody comprises VH CDR1, CDR2, and CDR3 domains comprising the amino acid sequences set forth in SEQ ID NOs: 1, 2, and 3, respectively, and a VL region comprising the amino acid sequence of SEQ ID NO: 10, but wherein:
 (i) the amino acid in the position corresponding to position T41 in SEQ ID NO: 10 is not T;   (ii) the amino acid in the position corresponding to position F10 in SEQ ID NO: 10 is not F, and wherein one or more of the amino acid positions corresponding to the positions T41, K55, and L97 in SEQ ID NO: 10 are not T, K, and L, respectively;   (iii) the amino acids in the positions corresponding to positions R23 and A35 in SEQ ID NO: 10 are not R and A, respectively;   (iv) the amino acids in the positions corresponding to positions F10, R23, A35, R47, D71, A82, D83, S86, 187, and F89 in SEQ ID NO: 10 are not F, R, A, R, D, A, D, S, I, and F, respectively;   (v) the amino acid in the position corresponding to position K55 in SEQ ID NO: 10 is not K; or   (vi) the amino acid in the position corresponding to position F10 in SEQ ID NO: 10 is not F, and the amino acid in the position corresponding to position K55 in SEQ ID NO: 10 is not K,   wherein the numbering of positions is according to the EU Index.   
     
     
         68 . The nucleic acid of  claim 67 , wherein in subpart (i), the amino acid in the position corresponding to position T41 in SEQ ID NO: 10 is selected from the group consisting of: H, I, K, L, Q, R, and V. 
     
     
         69 . The nucleic acid of  claim 67 , wherein in subpart (ii), the amino acids in the positions corresponding to positions F10, T41, K55, and L97 in SEQ ID NO: 10 are not F, T, K, and L, respectively. 
     
     
         70 . The nucleic acid of  claim 69 , wherein the amino acid in the position corresponding to position T41 in SEQ ID NO: 10 is selected from the group consisting of: H, I, K, L, Q, R or V, and wherein the amino acids in the positions corresponding to positions F10, K55, and L97 in SEQ ID NO: 10 are L, N, and H, respectively. 
     
     
         71 . The nucleic acid of  claim 67 , wherein in subpart (iii), the amino acids in the positions corresponding to positions R23 and A35 in SEQ ID NO: 10 are A and P, respectively. 
     
     
         72 . The nucleic acid of  claim 67 , wherein in subpart (iv), the amino acid in the position corresponding to position R23 in SEQ ID NO: 10 is selected from the group consisting of A, G, H, K, Q, S, and T, the amino acid in the position corresponding to A35 in SEQ ID NO: 10 is selected from the group consisting of I, L, M, P, V, G, F, and W, and the amino acids in the positions corresponding to positions F10, R47, D71, A82, D83, S86, 187, and F89 in SEQ ID NO: 10 are L, T, G, P, E, A, E, and Y, respectively. 
     
     
         73 . The nucleic acid of  claim 72 , wherein the amino acids in the positions corresponding to positions F10, R23, A35, R47, D71, A82, D83, S86, 187, and F89 in SEQ ID NO: 10 are L, A, P, T, G, P, E, A, E, and Y, respectively. 
     
     
         74 . The nucleic acid of  claim 67 , wherein in subpart (v), the amino acid in the position corresponding to position K55 in SEQ ID NO: 10 is N. 
     
     
         75 . The nucleic acid of  claim 67 , wherein in subpart (vi), the amino acid in the position corresponding to position F10 in SEQ ID NO: 10 is L, and the amino acid in the position corresponding to position K55 in SEQ ID NO: 10 is N. 
     
     
         76 . The nucleic acid of  claim 67 , wherein the VH region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 6-9. 
     
     
         77 . A nucleic acid encoding one or more amino acid sequences set forth in Table 1. 
     
     
         78 . An expression vector comprising the nucleic acid of  claim 67 . 
     
     
         79 . A host cell comprising the nucleic acid of  claim 67 . 
     
     
         80 . A nucleic acid encoding (a) a light chain variable (VL) region, (b) both heavy chain variable (VH) and VL regions, (c) a light chain, or (d) both heavy and light chains of a humanized or chimeric antibody which binds to human CD3, wherein the antibody comprises VH CDR1, CDR2, and CDR3 domains comprising the amino acid sequences set forth in SEQ ID NOs: 1, 2, and 3, respectively, and VL CDR1, CDR2, and CDR3 domains comprising the amino acid sequences set forth in SEQ ID NO: 4, GTN, and SEQ ID NO: 5 or 60, respectively,
 wherein (i) the amino acids in the positions corresponding to positions F10, T41, K55, and L97 in the VL region of SEQ ID NO: 10 are L, K, N, and H, respectively, or (ii) the amino acid in the position corresponding to position T41 in the VL region of SEQ ID NO: 10 is K, and   wherein said antibody comprises a first heavy chain and a second heavy chain and a first light chain and a second light chain, wherein the amino acids in the positions corresponding to positions L234, L235, and D265 in the human IgG1 heavy chain of SEQ ID NO: 15 are F, E, and A, respectively, in both the first heavy chain and second heavy chain, and wherein the amino acid in the position corresponding to F405 in the human IgG1 heavy chain of SEQ ID NO: 15 is L in the first heavy chain, and the amino acid in the position corresponding to K409 in the human IgG1 heavy chain of SEQ ID NO: 15 is R in the second heavy chain,   wherein the numbering of positions is according to the EU Index.   
     
     
         81 . The nucleic acid of  claim 80 , wherein the VH region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 6-9. 
     
     
         82 . An expression vector comprising the nucleic acid of  claim 81 . 
     
     
         83 . A host cell comprising the nucleic acid of  claim 81 . 
     
     
         84 . A host cell comprising the expression vector of  claim 82 . 
     
     
         85 . A nucleic acid comprising a nucleotide sequence encoding (a) a heavy chain and/or (b) a light chain of a humanized or chimeric antibody which binds to human CD3, wherein the antibody comprises VH CDR1, CDR2, and CDR3 domains comprising the amino acid sequences set forth in SEQ ID NOs: 1, 2, and 3, respectively, and a VL region comprising an amino acid sequence at least 95% identical to SEQ ID NO: 10, but wherein:
 (i) the amino acid in the position corresponding to position T41 in SEQ ID NO: 10 is not T;   (ii) the amino acid in the position corresponding to position F10 in SEQ ID NO: 10 is not F, and wherein one or more of the amino acid positions corresponding to the positions T41, K55, and L97 in SEQ ID NO: 10 are not T, K, and L, respectively;   (iii) the amino acids in the positions corresponding to positions R23 and A35 in SEQ ID NO: 10 are not R and A, respectively;   (iv) the amino acids in the positions corresponding to positions F10, R23, A35, R47, D71, A82, D83, S86, 187, and F89 in SEQ ID NO: 10 are not F, R, A, R, D, A, D, S, I, and F, respectively;   (v) the amino acid in the position corresponding to position K55 in SEQ ID NO: 10 is not K; or   (vi) the amino acid in the position corresponding to position F10 in SEQ ID NO: 10 is not F, and the amino acid in the position corresponding to position K55 in SEQ ID NO: 10 is not K, wherein the numbering of positions is according to the EU Index.   
     
     
         86 . A method for detecting the presence of CD3 antigen, or a cell expressing CD3 in a sample, comprising the steps of:
 (a) contacting the sample with a humanized or chimeric antibody which binds to human CD3, wherein the antibody comprises a heavy chain variable (VH) region comprising VH CDR1, CDR2, and CDR3 domains comprising the amino acid sequences set forth in SEQ ID NOs: 1, 2, and 3, respectively, and a light chain variable (VL) region comprising the amino acid sequence of SEQ ID NO: 10, but wherein:
 (i) the amino acid in the position corresponding to position T41 in SEQ ID NO: 10 is not T; 
 (ii) the amino acid in the position corresponding to position F10 in SEQ ID NO: 10 is not F, and wherein one or more of the amino acid positions corresponding to the positions T41, K55, and L97 in SEQ ID NO: 10 are not T, K, and L, respectively; 
 (iii) the amino acids in the positions corresponding to positions R23 and A35 in SEQ ID NO: 10 are not R and A, respectively; 
 (iv) the amino acids in the positions corresponding to positions F10, R23, A35, R47, D71, A82, D83, S86, 187, and F89 in SEQ ID NO: 10 are not F, R, A, R, D, A, D, S, I, and F, respectively; 
 (v) the amino acid in the position corresponding to position K55 in SEQ ID NO: 10 is not K; or 
 (vi) the amino acid in the position corresponding to position F10 in SEQ ID NO: 10 is not F, and the amino acid in the position corresponding to position K55 in SEQ ID NO: 10 is not K,
 wherein the numbering of positions is according to the EU Index, under conditions that allow for formation of a complex between the antibody and CD3; and 
 
   (b) analyzing whether a complex has been formed.