Compositions comprising cross-species-specific antibodies and uses thereof

Abstract

The present invention relates to uses of bispecific antibodies exhibiting cross-species specificity for evaluating the in vivo safety and/or activity and/or pharmacokinetic profile of the same in non-human species and humans. The present invention moreover relates to methods for evaluating the in vivo safety and/or activity and/or pharmacokinetic profile of said bispecific antibodies exhibiting cross-species specificity. The present invention also relates to methods of measuring the biological activity and/or efficacy of such bispecific antibodies exhibiting cross-species specificity. In addition, the present invention relates to pharmaceutical compositions comprising bispecific single chain antibodies exhibiting cross-species specificity and to methods for the preparation of pharmaceutical compositions comprising said bispecific single chain antibodies exhibiting cross-species specificity for the treatment of diseases.

1 . A bispecific single chain antibody which comprises
 (i) a first binding domain binding to an epitope of human and non-chimpanzee primate CD3, and   (ii) a second binding domain binding to a cell surface antigen,   wherein the epitope comprises the amino acid sequence “FSEXE” (SEQ ID NO. 204), wherein “X” represents L (Leucine) or M (Methionine).   
     
     
         2 . (canceled) 
     
     
         3 . The bispecific single chain antibody of  claim 1 , wherein said first binding domain is located C-terminally or N-terminally to the second binding domain. 
     
     
         4 . The bispecific single chain antibody of  claim 1 , wherein the second binding domain binds to a human cell surface antigen and to the non-chimpanzee primate homolog of said cell surface antigen. 
     
     
         5 . The bispecific single chain antibody of  claim 1 , wherein the cell surface antigen is a tumor antigen. 
     
     
         6 . The bispecific single chain antibody of  claim 1 , wherein the first binding domain comprises a VH region having an amino acid sequence as shown in any of SEQ ID NOs. 2, 110 or 6. 
     
     
         7 . The bispecific single chain antibody of  claim 1 , wherein the first binding domain comprises a VL region having an amino acid sequence as shown in any of SEQ ID NOs. 4, 148, 168 or 8. 
     
     
         8 . The bispecific single chain antibody of  claim 1 , wherein the VH region of the first binding domain comprises or consists of the amino acid sequence shown in SEQ ID NO. 2 and the VL region of the first binding domain comprises or consists of the amino acid sequence shown in SEQ ID NO. 4. 
     
     
         9 . The bispecific single chain antibody of  claim 1 , wherein the VH region of the first binding domain comprises or consists of the amino acid sequence shown in SEQ ID NO. 110 and the VL region of the first binding domain comprises or consists of the amino acid sequence shown in SEQ ID NO. 148 or SEQ ID NO: 168. 
     
     
         10 . The bispecific single chain antibody of  claim 1 , wherein the VH region of the first binding domain comprises or consists of the amino acid sequence shown in SEQ ID NO. 6 and the VL region of the first binding domain comprises or consists of the amino acid sequence shown in SEQ ID NO. 8. 
     
     
         11 . The bispecific single chain antibody of  claim 5 , wherein said tumor antigen is EpCAM, EGFR, EGFRvIII or Carboanhydrase IX (MN/CA IX). 
     
     
         12 . The bispecific single chain antibody of  claim 1 , wherein the non-chimpanzee primate is a baboon, marmoset or an old world monkey. 
     
     
         13 . The bispecific single chain antibody of  claim 12 , wherein the old world monkey is a monkey of the macaque genus. 
     
     
         14 . The bispecific single chain antibody of  claim 13 , wherein the monkey of the macaque genus is Assamese macaque ( Macaca assamensis ), Barbary macaque ( Macaca sylvanus ), Bonnet macaque ( Macaca radiata ), Booted or Sulawesi-Booted macaque ( Macaca ochreata ), Sulawesi-crested macaque ( Macaca nigra ), Formosan rock macaque ( Macaca cyclopsis ), Japanese snow macaque or Japanese macaque ( Macaca fuscata ), Cynomolgus monkey or crab-eating macaque or long-tailed macaque or Java macaque ( Macaca fascicularis ), Lion-tailed macaque ( Macaca silenus ), Pigtailed macaque ( Macaca nemestrina ), Rhesus macaque ( Macaca mulatta ), Tibetan macaque ( Macaca thibetana ), Tonkean macaque ( Macaca tonkeana ), Toque macaque ( Macaca sinica ), Stump-tailed macaque or Red-faced macaque or Bear monkey ( Macaca arctoides ), or Moor macaque ( Macaca maurus ). 
     
     
         15 . The bispecific single chain antibody of  claim 1 , wherein the non-chimpanzee primate CD3 comprises or consists of an amino acid sequence shown in SEQ ID NOs. 135, 136, 144 or 145. 
     
     
         16 . The bispecific single chain antibody of  claim 1 , wherein said bispecific single chain antibody comprises an amino acid sequence selected from the group consisting of:
 (a) an amino acid sequence as depicted in any of SEQ ID NOs. 38, 40, 124, 42 or 44;   (b) an amino acid sequence encoded by a nucleic acid sequence as shown in SEQ ID NOs. 37, 39, 125, 41 or 43;   (c) an amino acid sequence encoded by a nucleic acid sequence hybridising under stringent conditions to the complementary nucleic acid sequence of (b);   (d) an amino acid sequence encoded by a nucleic acid sequence which is degenerate as a result of the genetic code to a nucleotide sequence of (b); and   (e) an amino acid sequence at least 85% identical, more preferred at least 90% identical, most preferred at least 95% identical to the amino acid sequence of (a) or (b).   
     
     
         17 . The bispecific single chain antibody of  claim 1 , wherein said bispecific single chain antibody comprises an amino acid sequence selected from the group consisting of:
 (a) an amino acid sequence as depicted in any of SEQ ID NOs. 66, 68, 74, 76, 122, 70, 72, 78, 80, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, or 192;   (b) an amino acid sequence encoded by a nucleic acid sequence as shown in SEQ ID NOs. 65, 67, 73, 75, 123, 69, 71, 77, 79, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, or 191;   (c) an amino acid sequence encoded by a nucleic acid sequence hybridising under stringent conditions to the complementary nucleic acid sequence of (b);   (d) an amino acid sequence encoded by a nucleic acid sequence which is degenerate as a result of the genetic code to a nucleotide sequence of (b); and   (e) an amino acid sequence at least 85% identical, more preferred at least 90% identical, most preferred at least 95% identical to the amino acid sequence of (a) or (b).   
     
     
         18 . The bispecific single chain antibody of  claim 1 , wherein at least one of said first or second binding domains is human, humanized, CDR-grafted and/or deimmunized. 
     
     
         19 . An isolated nucleic acid sequence encoding the bispecific single chain antibody of  claim 1 . 
     
     
         20 . A vector which comprises a nucleic acid sequence of  claim 19 , wherein said vector optionally comprises a regulatory sequence which is operably linked to said nucleic acid sequence. 
     
     
         21 .- 22 . (canceled) 
     
     
         23 . A host transformed or transfected with said nucleic acid sequence of  claim 19  or a vector comprising said nucleic acid sequence. 
     
     
         24 . The pharmaceutical composition of  claim 26 , further comprising a proteinaceous compound capable of providing an activation signal for immune effector cells. 
     
     
         25 . A process for the production of a bispecific single chain antibody comprising culturing a host transformed or transfected with a nucleic acid sequence encoding the bispecific single chain antibody of  claim 1  or a vector comprising said nucleic acid sequence, under conditions allowing the expression of the bispecific single chain antibody, and recovering the produced bispecific single chain antibody from the culture. 
     
     
         26 . A pharmaceutical composition comprising the bispecific single chain antibody of  claim 1 , which is further comprises suitable formulations of carriers, stabilizers and/or excipients. 
     
     
         27 .- 30 . (canceled) 
     
     
         31 . A method for the prevention, treatment or amelioration of a disease in a subject in the need thereof, said method comprising the administration of an effective amount of a pharmaceutical composition of  claim 26 . 
     
     
         32 . The method of  claim 31 , wherein said disease is a proliferative disease, a tumorous disease, or an immunological disorder. 
     
     
         33 . The method of  claim 32 , wherein said tumorous disease is a malignant disease, preferably cancer. 
     
     
         34 . The method of  claim 31 , wherein said pharmaceutical composition comprising the bispecific single chain antibody is administered in combination with an additional drug. 
     
     
         35 . The method of  claim 34 , wherein said drug is a non-proteinaceous compound or a proteinaceous compound. 
     
     
         36 . The method of  claim 35 , comprising the administration of a proteinaceous compound capable of providing an activation signal for immune effector cells. 
     
     
         37 . The method of  claim 35 , wherein said proteinaceous compound or non-proteinaceous compound is administered simultaneously or non-simultaneously with the pharmaceutical composition comprising the bispecific single chain antibody. 
     
     
         38 . The method of  claim 31 , wherein said subject is a human. 
     
     
         39 . A kit comprising a bispecific single chain antibody of  claim 1 . 
     
     
         40 .- 43 . (canceled)