Antibodies

Abstract

The present invention provides antibodies which bind to CXC chemokine receptor 4 (CXCR4) and which do not induce significant apoptosis of CXCR4 expressing cells. Also provided are inter alia immunoconjugates and compositions comprising such antibodies and methods and uses involving such antibodies, particularly in the medical and diagnostic fields.

1 . A nucleic acid molecule comprising a nucleotide sequence region encoding:
 (A) an isolated antibody which binds to CXC chemokine receptor 4 (CXCR4) and which does not induce significant apoptosis of CXCR4 expressing cells, wherein by the term not inducing significant apoptosis of CXCR4 expressing cells is meant that levels of apoptosis induced in the presence of an antibody are comparable to or not significantly different from levels of apoptosis induced in the absence of an antibody, and wherein said antibody comprises at least one heavy chain variable region that comprises three CDRs and at least one light chain variable region that comprises three CDRs,   wherein said heavy chain variable region comprises:
 (i) a variable heavy (VH) CDR1 that has the amino acid sequence of S/G Y X 3  M/I H/S (SEQ ID NO: 126), or X 1  Y X 3  M H (SEQ ID NO: 127) or S Y X 3  M H (SEQ ID NO: 128); 
   wherein X 1  can be S or G, preferably S;   X 3  can be G or W or Y or A, preferably W;
 (ii) a VH CDR2 that has the amino acid sequence of X 1  I X 3  X 4  D G S X 8  X 9  X 10  Y AD S V K G (SEQ ID NO:129); 
   wherein X 1  can be V or R, preferably R;   X 3  can be S or N, preferably N;   X 4  can be Y or S, preferably S;   X 8  can be N or S, preferably S;   X 9  can be K or T, preferably T; and   X 10  can be Y or S, preferably S;
 or a VH CDR2 that has the amino acid sequence of X 1  I X 3  P X 5  X 6  G X 8  X 9  N Y A Q K F Q G (SEQ ID NO: 131); 
   wherein X 1 , can be R or G, preferably R;   X 3  can be N or I, preferably N;   X 5  can be N or I, preferably N;   X 6  can be S or F, preferably S;   X 8  can be G or T, preferably G; and   X 9  can be T or A, preferably T;   and
 (iii) a VH CDR3 that has the amino acid sequence of SEQ ID NOs: 3, 9, 15 or 21; or 
   (B) a heavy chain variable region as defined in part (A); or   (C) a light chain variable region that comprises three CDRs,
 wherein said light chain variable region comprises:
 (i) a variable light (VL) CDR1 that has the amino acid sequence of SEQ ID NO: 4; 
 (ii) a VL CDR2 that has the amino acid sequence of SEQ ID NO: 5; and 
 (iii) a VL CDR3 that has the amino acid sequence of SEQ ID NO: 6; or 
 
 wherein said light chain variable region comprises:
 (i) a variable light (VL) CDR1 that has the amino acid sequence of SEQ ID NO: 10; 
 (ii) a VL CDR2 that has the amino acid sequence of SEQ ID NO: 11; and 
 (iii) a VL CDR3 that has the amino acid sequence of SEQ ID NO: 12; or 
 
 wherein said light chain variable region comprises:
 (i) a variable light (VL) CDR1 that has the amino acid sequence of SEQ ID NO: 16; 
 (ii) a VL CDR2 that has the amino acid sequence of SEQ ID NO: 17; and 
 (iii) a VL CDR3 that has the amino acid sequence of SEQ ID NO: 18; or 
 
 wherein said light chain variable region comprises:
 (i) a variable light (VL) CDR1 that has the amino acid sequence of SEQ ID NO: 22; 
 (ii) a VL CDR2 that has the amino acid sequence of SEQ ID NO: 23; and 
 (iii) a VL CDR3 that has the amino acid sequence of SEQ ID NO: 24; or 
 
 wherein said light chain variable region comprises:
 (i) a variable light (VL) CDR1 that has the amino acid sequence of SEQ ID NO: 88; 
 (ii) a VL CDR2 that has the amino acid sequence of SEQ ID NO: 89; and 
 (iii) a VL CDR3 that has the amino acid sequence of SEQ ID NO: 90. 
 
   
     
     
         2 . The nucleic acid molecule according to  claim 1 , wherein said heavy chain variable region comprises:
 (i) a variable heavy (VH) CDR1 that has the amino acid sequence of SEQ ID NO: 1;   (ii) a VH CDR2 that has the amino acid sequence of SEQ ID NO: 2; and   (iii) a VH CDR3 that has the amino acid sequence of SEQ ID NO: 3; or
 wherein said heavy chain variable region comprises:
 (i) a variable heavy (VH) CDR1 that has the amino acid sequence of SEQ ID NO: 7; 
 (ii) a VH CDR2 that has the amino acid sequence of SEQ ID NO: 8; and 
 (iii) a VH CDR3 that has the amino acid sequence of SEQ ID NO: 9; or 
 
 wherein said heavy chain variable region comprises:
 (i) a variable heavy (VH) CDR1 that has the amino acid sequence of SEQ ID NO: 13; 
 (ii) a VH CDR2 that has the amino acid sequence of SEQ ID NO: 14; and 
 (iii) a VH CDR3 that has the amino acid sequence of SEQ ID NO: 15; or 
 
 wherein said heavy chain variable region comprises:
 (i) a variable heavy (VH) CDR1 that has the amino acid sequence of SEQ ID NO: 19; 
 (ii) a VH CDR2 that has the amino acid sequence of SEQ ID NO: 20; and 
 (iii) a VH CDR3 that has the amino acid sequence of SEQ ID NO: 21. 
 
   
     
     
         3 . The nucleic acid molecule according to  claim 1 , wherein said antibody comprises at least one heavy chain variable region that comprises three CDRs and at least one light chain variable region that comprises three CDRs,
 wherein said heavy chain variable region comprises:
 (i) a variable heavy (VH) CDR1 that has the amino acid sequence of SEQ ID NO: 1; 
 (ii) a VH CDR2 that has the amino acid sequence of SEQ ID NO: 2; and 
 (iii) a VH CDR3 that has the amino acid sequence of SEQ ID NO: 3; and/or 
   wherein said light chain variable region comprises:
 (iv) a variable light (VL) CDR1 that has the amino acid sequence of SEQ ID NO: 4; 
 (v) a VL CDR2 that has the amino acid sequence of SEQ ID NO: 5; and 
 (vi) a VL CDR3 that has the amino acid sequence of SEQ ID NO: 6. 
   
     
     
         4 . The nucleic acid molecule according to  claim 1 , wherein said antibody comprises at least one heavy chain variable region that comprises three CDRs and at least one light chain variable region that comprises three CDRs,
 wherein said heavy chain variable region comprises:
 (i) a variable heavy (VH) CDR1 that has the amino acid sequence of SEQ ID NO: 7; 
 (ii) a VH CDR2 that has the amino acid sequence of SEQ ID NO: 8; and 
 (iii) a VH CDR3 that has the amino acid sequence of SEQ ID NO: 9; and/or 
   wherein said light chain variable region comprises:
 (iv) a variable light (VL) CDR1 that has the amino acid sequence of SEQ ID NO: 10; 
 (v) a VL CDR2 that has the amino acid sequence of SEQ ID NO: 11; and 
 (vi) a VL CDR3 that has the amino acid sequence of SEQ ID NO: 12. 
   
     
     
         5 . The nucleic acid molecule according to  claim 1 , wherein said antibody comprises at least one heavy chain variable region that comprises three CDRs and at least one light chain variable region that comprises three CDRs,
 wherein said heavy chain variable region comprises:
 (i) a variable heavy (VH) CDR1 that has the amino acid sequence of SEQ ID NO: 13; 
 (ii) a VH CDR2 that has the amino acid sequence of SEQ ID NO: 14; and 
 (iii) a VH CDR3 that has the amino acid sequence of SEQ ID NO: 15; and/or 
   wherein said light chain variable region comprises:
 (iv) a variable light (VL) CDR1 that has the amino acid sequence of SEQ ID NO: 16; 
 (v) a VL CDR2 that has the amino acid sequence of SEQ ID NO: 17; and 
 (vi) a VL CDR3 that has the amino acid sequence of SEQ ID NO: 18. 
   
     
     
         6 . The nucleic acid molecule according to  claim 1 , wherein said antibody comprises at least one heavy chain variable region that comprises three CDRs and at least one light chain variable region that comprises three CDRs,
 wherein said heavy chain variable region comprises:
 (i) a variable heavy (VH) CDR1 that has the amino acid sequence of SEQ ID NO: 19; 
 (ii) a VH CDR2 that has the amino acid sequence of SEQ ID NO: 20; and 
 (iii) a VH CDR3 that has the amino acid sequence of SEQ ID NO: 21; and/or 
   wherein said light chain variable region comprises:
 (iv) a variable light (VL) CDR1 that has the amino acid sequence of SEQ ID NO: 22; 
 (v) a VL CDR2 that has the amino acid sequence of SEQ ID NO: 23; and 
 (vi) a VL CDR3 that has the amino acid sequence of SEQ ID NO: 24. 
   
     
     
         7 . The nucleic acid molecule according to  claim 1 , wherein said antibody comprises at least one heavy chain variable region that comprises three CDRs and at least one light chain variable region that comprises three CDRs,
 wherein said heavy chain variable region comprises:
 (i) a variable heavy (VH) CDR1 that has the amino acid sequence of SEQ ID NO: 1; 
 (ii) a VH CDR2 that has the amino acid sequence of SEQ ID NO: 2; and 
 (iii) a VH CDR3 that has the amino acid sequence of SEQ ID NO: 3; and/or 
   wherein said light chain variable region comprises:
 (iv) a variable light (VL) CDR1 that has the amino acid sequence of SEQ ID NO: 88; 
 (v) a VL CDR2 that has the amino acid sequence of SEQ ID NO: 89; and 
 (vi) a VL CDR3 that has the amino acid sequence of SEQ ID NO: 90. 
   
     
     
         8 . The nucleic acid molecule according to  claim 3 , wherein said heavy chain variable region comprises one of each of the VH CDR domains (i), (ii) and (iii), and said light chain variable region comprises one of each of the VL CDR domains (iv), (v) and (vi). 
     
     
         9 . The nucleic acid molecule according to  claim 4 , wherein said heavy chain variable region comprises one of each of the VH CDR domains (i), (ii) and (iii), and said light chain variable region comprises one of each of the VL CDR domains (iv), (v) and (vi). 
     
     
         10 . The nucleic acid molecule according to  claim 5 , wherein said heavy chain variable region comprises one of each of the VH CDR domains (i), (ii) and (iii), and said light chain variable region comprises one of each of the VL CDR domains (iv), (v) and (vi). 
     
     
         11 . The nucleic acid molecule according to  claim 6 , wherein said heavy chain variable region comprises one of each of the VH CDR domains (i), (ii) and (iii), and said light chain variable region comprises one of each of the VL CDR domains (iv), (v) and (vi). 
     
     
         12 . The nucleic acid molecule according to  claim 7 , wherein said heavy chain variable region comprises one of each of the VH CDR domains (i), (ii) and (iii), and said light chain variable region comprises one of each of the VL CDR domains (iv), (v) and (vi). 
     
     
         13 . The nucleic acid molecule according to  claim 1 , wherein (i) said heavy chain variable region has an amino acid sequence of SEQ ID NO: 69, preferably encoded by SEQ ID NO:77, or wherein (ii) said light chain variable region has an amino acid sequence of SEQ ID NO: 70, preferably encoded by SEQ ID NO:78, or wherein (iii) said heavy chain variable region has an amino acid sequence of SEQ ID NO: 69, preferably encoded by SEQ ID NO:77, and said light chain variable region has an amino acid sequence of SEQ ID NO: 70, preferably encoded by SEQ ID NO:78. 
     
     
         14 . The nucleic acid molecule according to  claim 1 , wherein (i) said heavy chain variable region has an amino acid sequence of SEQ ID NO: 71, preferably encoded by SEQ ID NO:79, or wherein (ii) said light chain variable region has an amino acid sequence of SEQ ID NO: 72, preferably encoded by SEQ ID NO:80, or wherein (iii) said heavy chain variable region has an amino acid sequence of SEQ ID NO: 71, preferably encoded by SEQ ID NO:79, and said light chain variable region has an amino acid sequence of SEQ ID NO: 72, preferably encoded by SEQ ID NO:80. 
     
     
         15 . The nucleic acid molecule according to  claim 1 , wherein (i) said heavy chain variable region has an amino acid sequence of SEQ ID NO: 73, preferably encoded by SEQ ID NO:81, or wherein (ii) said light chain variable region has an amino acid sequence of SEQ ID NO:74, preferably encoded by SEQ ID NO:82, or wherein (iii) said heavy chain variable region has an amino acid sequence of SEQ ID NO: 73, preferably encoded by SEQ ID NO:81, and said light chain variable region has an amino acid sequence of SEQ ID NO: 74, preferably encoded by SEQ ID NO:82. 
     
     
         16 . The nucleic acid molecule according to  claim 1 , wherein (i) said heavy chain variable region has an amino acid sequence of SEQ ID NO: 75, preferably encoded by SEQ ID NO:83, or wherein (ii) said light chain variable region has an amino acid sequence of SEQ ID NO:76, preferably encoded by SEQ ID NO:84, or wherein (iii) said heavy chain variable region has an amino acid sequence of SEQ ID NO: 75, preferably encoded by SEQ ID NO:83, and said light chain variable region has an amino acid sequence of SEQ ID NO: 76, preferably encoded by SEQ ID NO:84. 
     
     
         17 . The nucleic acid molecule according to  claim 1 , wherein (i) said heavy chain variable region has an amino acid sequence of SEQ ID NO: 69, preferably encoded by SEQ ID NO:77, or wherein (ii) said light chain variable region has an amino acid sequence of SEQ ID NO:103, preferably encoded by SEQ ID NO:105, or wherein (iii) said heavy chain variable region has an amino acid sequence of SEQ ID NO: 69, preferably encoded by SEQ ID NO:77, and said light chain variable region has an amino acid sequence of SEQ ID NO: 103, preferably encoded by SEQ ID NO:105. 
     
     
         18 . The nucleic acid molecule according to  claim 1 , wherein said antibody comprises the amino acid sequence of SEQ ID NO:35, SEQ ID NO:46, SEQ ID NO:57, SEQ ID NO:68 or SEQ ID NO:101. 
     
     
         19 . The nucleic acid molecule according to  claim 1 , wherein said nucleotide sequence region has the nucleotide sequence of SEQ ID NO: 34, SEQ ID NO: 45, SEQ ID NO: 56, SEQ ID NO: 67 or SEQ ID NO: 100. 
     
     
         20 . The nucleic acid molecule according to  claim 1 , wherein (i) said nucleotide sequence region encodes a heavy chain of an antibody that comprises an amino acid sequence of SEQ ID NO:108, preferably encoded by SEQ ID NO:106, or wherein (ii) said nucleotide sequence region encodes a light chain of an antibody that comprises an amino acid sequence of SEQ ID NO:109, preferably encoded by SEQ ID NO:107, or wherein (iii) said nucleotide sequence region encodes a heavy chain of an antibody that comprises an amino acid sequence of SEQ ID NO: 108, preferably encoded by SEQ ID NO:106, and a light chain of an antibody that comprises an amino acid sequence of SEQ ID NO: 109, preferably encoded by SEQ ID NO:107. 
     
     
         21 . The nucleic acid molecule according to  claim 1 , wherein (i) said nucleotide sequence region encodes a heavy chain of an antibody that comprises an amino acid sequence of SEQ ID NO:112, preferably encoded by SEQ ID NO:110, or wherein (ii) said nucleotide sequence region encodes a light chain of an antibody that comprises an amino acid sequence of SEQ ID NO:113, preferably encoded by SEQ ID NO:111, or wherein (iii) said nucleotide sequence region encodes a heavy chain of an antibody that comprises an amino acid sequence of SEQ ID NO:112, preferably encoded by SEQ ID NO:110, and a light chain of an antibody that comprises an amino acid sequence of SEQ ID NO:113, preferably encoded by SEQ ID NO:111. 
     
     
         22 . The nucleic acid molecule according to  claim 1 , wherein (i) said nucleotide sequence region encodes a heavy chain of an antibody that comprises an amino acid sequence of SEQ ID NO:116, preferably encoded by SEQ ID NO:114, or wherein (ii) said nucleotide sequence region encodes a light chain of an antibody that comprises an amino acid sequence of SEQ ID NO:117, preferably encoded by SEQ ID NO:115, or wherein (iii) said nucleotide sequence region encodes a heavy chain of an antibody that comprises an amino acid sequence of SEQ ID NO:116, preferably encoded by SEQ ID NO:114, and a light chain of an antibody that comprises an amino acid sequence of SEQ ID NO:117, preferably encoded by SEQ ID NO:115. 
     
     
         23 . The nucleic acid molecule according to  claim 1 , wherein (i) said nucleotide sequence region encodes a heavy chain of an antibody that comprises an amino acid sequence of SEQ ID NO:120, preferably encoded by SEQ ID NO:118, or wherein (ii) said nucleotide sequence region encodes a light chain of an antibody that comprises an amino acid sequence of SEQ ID NO:121, preferably encoded by SEQ ID NO:119, or wherein (iii) said nucleotide sequence region encodes a heavy chain of an antibody that comprises an amino acid sequence of SEQ ID NO:120, preferably encoded by SEQ ID NO:118, and a light chain of an antibody that comprises an amino acid sequence of SEQ ID NO:121, preferably encoded by SEQ ID NO:119. 
     
     
         24 . The nucleic acid molecule according to  claim 1 , wherein (i) said nucleotide sequence region encodes a heavy chain of an antibody that comprises an amino acid sequence of SEQ ID NO:108, preferably encoded by SEQ ID NO:106, or wherein (ii) said nucleotide sequence region encodes a light chain of an antibody that comprises an amino acid sequence of SEQ ID NO:125, preferably encoded by SEQ ID NO:123, or wherein (iii) said nucleotide sequence region encodes a heavy chain of an antibody that comprises an amino acid sequence of SEQ ID NO:108, preferably encoded by SEQ ID NO:106, and a light chain of an antibody that comprises an amino acid sequence of SEQ ID NO:125, preferably encoded by SEQ ID NO:123. 
     
     
         25 . An expression vector comprising a nucleic acid molecule of  claim 1 . 
     
     
         26 . A host cell comprising one or more nucleic acid molecules of  claim 1  or one or more expression vectors of  claim 25 . 
     
     
         27 . A virus comprising one or more nucleic acid molecules of  claim 1  or one or more expression vectors of  claim 25 . 
     
     
         28 . A method of producing an antibody, comprising:
 (a) culturing a host cell that comprises one or more nucleic acid molecules of  claim 1  or one or more expression vectors of  claim 25  under conditions effective to express the encoded antibody; and   (b) obtaining the expressed antibody from said host cell;   wherein said antibody is an isolated antibody which binds to CXC chemokine receptor 4 (CXCR4) and which does not induce significant apoptosis of CXCR4 expressing cells, wherein by the term not inducing significant apoptosis of CXCR4 expressing cells is meant that levels of apoptosis induced in the presence of an antibody are comparable to or not significantly different from levels of apoptosis induced in the absence of an antibody, and wherein said antibody comprises at least one heavy chain variable region that comprises three CDRs and at least one light chain variable region that comprises three CDRs,   wherein said heavy chain variable region comprises:
 (i) a variable heavy (VH) CDR1 that has the amino acid sequence of S/G Y X 3  M/I H/S (SEQ ID NO: 126), or X 1  Y X 3  M H (SEQ ID NO: 127) or S Y X 3  M H (SEQ ID NO: 128); 
   wherein X 1  can be S or G, preferably S;   X 3  can be G or W or Y or A, preferably W;
 (ii) a VH CDR2 that has the amino acid sequence of X 1  I X 3  X 4  D G S X 8  X 9  X 10  Y A D S V K G (SEQ ID NO:129); 
   wherein X 1  can be V or R, preferably R;   X 3  can be S or N, preferably N;   X 4  can be Y or S, preferably S;   X 8  can be N or S, preferably S;   X 9  can be K or T, preferably T; and   X 10  can be Y or S, preferably S;
 or a VH CDR2 that has the amino acid sequence of X 1  I X 3  P X 5  X 6  G X 8  X 9  N Y A Q K F Q G (SEQ ID NO: 131); 
   wherein X 1 , can be R or G, preferably R;   X 3  can be N or I, preferably N;   X 5  can be N or I, preferably N;   X 6  can be S or F, preferably S;   X 8  can be G or T, preferably G; and   X 9  can be T or A, preferably T;   and
 (iii) a VH CDR3 that has the amino acid sequence of SEQ ID NOs: 3, 9, 15 or 21. 
   
     
     
         29 . The method of  claim 28 , further comprising a step of formulating the antibody into a pharmaceutical composition.