Composition Comprising Cross-Species-Sepecific Antibodies and uses Thereof

• US 20090252683A1
• Filed: 10/10/2006
• Published: 10/08/2009
• Est. Priority Date: 10/11/2005
• Status: Active Grant

• Current Assignee: Amgen Research (Munich) GmbH (Amgen Inc.)
• Original Assignee: Micromet AG (Amgen Inc.)
• Inventors: Cierpka, Ronny, Raum, Tobias, Kufer, Peter, Kischel, Roman, Rau, Doris, Schlereth, Bernd
• Granted Patent: US 8,236,308 B2
• Time in Patent Office: Days
• US Class Current: 424/9.200
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 39/39558 against tumor tissues, cells, antigens
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 16/2809 against the T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 16/30 from tumour cells
• CPC Class Codes: C07K 16/40 against enzymes
• CPC Class Codes: C07K 16/468 Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
• CPC Class Codes: C07K 2317/31 multispecific
• CPC Class Codes: C07K 2317/34 Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)

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 anti-bodies 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.

Claims

1. A pharmaceutical composition for the treatment of a human patient, comprising a bispecific single chain antibody which comprises

(i) a first binding domain binding to a non-chimpanzee primate CD3, and

(ii) a second binding domain binding to a cell surface antigen,

wherein said first binding domain binds to human and non-chimpanzee primate CD3.

2. The pharmaceutical composition of claim 1, wherein said first binding domain binds to an epitope of human and non-chimpanzee primate CD3 comprising the amino acid sequence “

FSEXE”

(SEQ ID NO.

     204), wherein “

X”

represents L (Leucine) or M (Methionine).

3. The pharmaceutical composition of claim 1, wherein said first binding domain is located C-terminally or N-terminally to the second binding domain.

4. The pharmaceutical composition 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 pharmaceutical composition of claim 1, wherein the cell surface antigen is a tumor antigen.

6. The pharmaceutical composition 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 pharmaceutical composition 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 pharmaceutical composition 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 pharmaceutical composition 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 pharmaceutical composition 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 pharmaceutical composition of claim 5, wherein said tumor antigen is EpCAM, EGFR, EGFRvIII or Carboanhydrase IX (MN/CA IX).

12. The pharmaceutical composition of claim 1, wherein the non-chimpanzee primate is a baboon, marmoset or an old world monkey.

13. The pharmaceutical composition of claim 12, wherein the old world monkey is a monkey of the macaque genus.

14. The pharmaceutical composition 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 pharmaceutical composition 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 pharmaceutical composition 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 pharmaceutical composition 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 pharmaceutical composition of claim 1, wherein at least one of said first or second binding domains is human, humanized, CDR-grafted and/or deimmunized.

19. A pharmaceutical composition comprising a nucleic acid sequence encoding a bispecific single chain antibody as defined in claim 1.

20. A pharmaceutical composition comprising a vector which comprises a nucleic acid sequence encoding a bispecific single chain antibody as defined in claim 1.

21. The pharmaceutical composition of claim 20, wherein said vector further comprises a regulatory sequence which is operably linked to said nucleic acid sequence.

22. The pharmaceutical composition of claim 20, wherein said vector is an expression vector.

23. A pharmaceutical composition comprising a host transformed or transfected with a vector defined in any of claim 20.

24. A pharmaceutical composition according to claim 1, further comprising a proteinaceous compound capable of providing an activation signal for immune effector cells.

25. A process for the production of a pharmaceutical composition according to claim 1, said process comprising culturing a host transformed or transfected with a vector which comprises a nucleic acid sequence encoding a bispecific single chain antibody as defined in claims 1, under conditions allowing the expression of the bispecific single chain antibody as defined in claim 1 and recovering the produced bispecific single chain antibody from the culture.

26. The pharmaceutical composition of claim 1, which further comprises suitable formulations of carriers, stabilizers and/or excipients.

31. A method for the prevention, treatment or amelioration of a disease in a subject in the need thereof, said method comprising the step of administration of an effective amount of a pharmaceutical composition of claim 1 or as produced according by a process comprising culturing a host transformed or transfected with a vector which comprises a nucleic acid sequence encoding a bispecific single chain antibody as defined in claims 1, under conditions allowing the expression of the bispecific single chain antibody as defined in claim 1 and recovering the produced bispecific single chain antibody from the culture.

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 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. A method for the prevention, treatment or amelioration of a disease in a subject in the need thereof said method comprising the step of administration of an effective amount of a pharmaceutical composition of claim 1 or as produced according by a process comprising culturing a host transformed or transfected with a vector which comprises a nucleic acid sequence encoding a bispecific single chain antibody as defined in claim 1, under conditions allowing the expression of the bispecific single chain antibody as defined in claim 1 and recovering the produced bispecific single chain antibody from the culture,

wherein said bispecific single chain antibody is administered in combination with an additional drug, wherein said drug is a proteinaceous or -non-proteinaceous compound and

wherein said proteinaceous compound or non-proteinaceous compound is administered simultaneously or non-simultaneously with a bispecific single chain antibody as defined in claim 1, a nucleic acid molecule encoding a bispecific single chain antibody as defined in claim 1, a vector which comprises a nucleic acid sequence encoding a bispecific single chain antibody as defined in claim 1, or a host transformed or transfected with a vector which comprises a nucleic acid sequence encoding a bispecific single chain antibody as defined in claims 1.

38. The method of claim 31, wherein said subject is a human.

39. A kit comprising a bispecific single chain antibody as defined in claim 1, a nucleic acid molecule encoding a bispecific single chain antibody as defined in claim 1, a vector which comprises a nucleic acid sequence encoding a bispecific single chain antibody as defined in claim 1, or a host transformed or transfected with a vector which comprises a nucleic acid sequence encoding a bispecific single chain antibody as defined in claims 1.

40. A method of determining the biological activity and/or efficacy of a bispecific single chain antibody as defined in claim 1, wherein said bispecific single chain antibody is administered to a non-chimpanzee primate and the in vivo activity is measured.

42. A method for evaluating the biological activity/safety/toxicity of a bispecific single chain antibody as defined in claim 1, comprising

(i) administering said bispecific single chain antibody to a non-chimpanzee primate,

(ii) measuring the in vivo safety and/or activity and/or pharmacokinetic profile of said bispecific single chain antibody in said non-human primate,

(iii) evaluating the in vivo safety and/or activity and/or pharmacokinetic profile of said bispecific single chain antibody in the non-human primate, and

(iv) determining an effective and non-toxic dose of said bispecific single chain antibody and administering said dose to humans.

43. The method of claim 40, wherein said in vivo activity is T cell activation, tumor target cell depletion, cytotoxicity, toxicity, occurrence of adverse side effects, and/or cytokine release.

27. (canceled)

28. (canceled)

29. (canceled)

30. (canceled)

41. (canceled)