Binding molecules and methods of use thereof

1. An isolated binding molecule of the formula (II):

wherein

at least one Z is Ab, wherein Ab is a polypeptide comprising at least one antigen binding site;

L is an independently selected linker moiety for each occurrence or B when adjacent to a Z which is Ab;

B is a bridging moiety, wherein B is linked to Ab through a carboxylate or amino linkage;

R is selected independently for each occurrence from the group consisting of alkyl, alkenyl, alkynyl, acyl, and hydrogen;

Z is an independently selected drug moiety, affinity moiety, tag moiety, pharmokinetic moiety, hydrogen, amino acid side chain moiety, or Ab for each occurrence;

w and y are each independently selected for each occurrence from the group consisting of 1, 2, 3, 4, and 5;

b and d are each independently selected for each occurrence from integers greater than 1, and pharmaceutically acceptable salts, esters, and prodrugs thereof.

2. The binding molecule of claim 1 , wherein at least one L is an attachment moiety.

3. The binding molecule of claim 1 , wherein y is 2.

4. The binding molecule of claim 1 , wherein d is 1, 2, or 3.

5. The binding molecule of claim 1 , wherein w is 1 or 2.

6. The binding molecule of claim 1 , wherein each R is independently alkyl or hydrogen.

7. The binding molecule of claim 1 , wherein at least one Z is a drug moiety.

8. The binding molecule of claim 7 , wherein said molecule comprises two or more drug moieties.

9. The binding molecule of claim 7 , wherein said drug moiety is an anti-cancer, antibiotic, or anti-inflammatory agent.

10. The binding molecule of claim 9 , wherein Z is an anticancer agent.

11. The binding molecule of claim 10 , wherein Z is doxorubicin, etoposide, taxane, paclitaxel, fluorouracyl, mitomycin, camptothecin, a vinca alkaloid, geldanamycin, a gemcitabine, geldanamycin, epothilone, cephalostatin, tubulin inhibitors, proteasome inhibitors, neocarzinostatin, calicheamicin, maytanisinoids, (RS)-cyclophophamide, 6-mercaptopurin, auristatin E, daunorubicin, or a derivative or analog thereof.

12. The binding molecule of claim 11 , wherein Z is a maytansinoid of the formula (IV):

wherein

R Z1 is halogen or hydrogen; and

R Z2 and R Z3 are each hydrogen or lower alkyl.

13. The binding molecule of claim 12 , wherein R Z1 is chlorine and R Z2 and R Z3 are each methyl.

14. The binding molecule of claim 11 , wherein Z is a taxane derivative of the formula (V):

wherein:

R T1 , R T2 , and R T3 and R T3 are each independently hydrogen, an electron withdrawing group, or an electron donating group;

R T4 , R T5 , R T6 are each independently a covalent bond to L, hydrogen, heterocyclic, an ester, an ether, a carbamate of the formula —CONR T10 R T11 , wherein R T10 and R T11 are each independently hydrogen, alkyl, alkenyl, alkynyl, acyl or aryl, provided that one of R T4 , R T5 , and R T6 is a covalent bond to L;

R T7 is alkyl, alkenyl, alkynyl, acyl or aryl; and

R T8 is alkoxy or aryl.

15. The binding molecule of claim 11 , wherein Z is a doxorubicin derivative of the formula (VI):

wherein:

Y is O or NR D5 , wherein R D5 is alkyl or hydrogen;

R D1 and R D2 are each hydrogen, or taken together a moiety of the formula (VIa):

R D3 is alkyl;

R D4 is alkyl or hydrogen;

R D6 is hydroxy or alkyl;

R D7 is O or a covalent bond to L;

R D8 and R D9 are each a covalent bond to L, hydrogen, or alkyl;

provided that one of R D1 , R D2 , and R D7 is a covalent bond to L.

16. The binding molecule of claim 1 , wherein at least one Z is an affinity moiety.

17. The binding molecule of claim 16 , wherein said affinity moiety is biotin.

18. The binding molecule of claim 1 , wherein at least one Z is a tag moiety.

19. The binding molecule of claim 18 , wherein said tag moiety is a fluorescent or radioactive tag.

20. The binding molecule of claim 1 , wherein at least one L is cleavable.

21. The binding molecule of claim 20 , wherein at least one L is selected such that it is cleaved extracellularly.

22. The binding molecule of claim 20 , wherein at least one L is selected such that it is cleaved intracellularly.

23. The binding molecule of claim 20 , wherein at least one L is cleaved by a drop of pH, enzymatic cleavage or a change in redox potential.

24. The binding molecule of claim 20 , wherein at least one L comprises a disulfide, acetal, ketal, orthoester, ester, trityl, cis-aconityl, thiocarbamoyl, or a peptide moiety.

25. The binding molecule of claim 1 , wherein at least one L is of the formula (VII):

(CR L1 R L2 ) f —NR L3 —C(═O)—(CR L4 R L5 ) g —S-D  (VII)

wherein

R L1 , R L2 , R L3 , R L4 , and R L5 are each independently alkyl, alkenyl, alkynyl, acyl, or hydrogen;

f and g are each independently selected for each occurrence from the group consisting of 0, 1, 2, 3, 4, 5, and 6; and

D is a drug attachment moiety.

26. The binding molecule of claim 25 , wherein R L1 , R L2 , R L3 , R L4 , and R L5 are each independently hydrogen or methyl.

27. The binding molecule of claim 25 , wherein f is 3 and each of R L1 and R L2 are hydrogen.

28. The binding molecule of claim 25 , wherein R L3 is hydrogen.

29. The binding molecule of claim 25 , wherein D comprises a moiety of the formula (VIII):

wherein

R V4 and R V5 are each hydrogen or lower alkyl; and

K is an alkyl or cycloalkyl linker comprising 1 to 10 carbon atoms.

30. The binding molecule of claim 25 , wherein D comprises a moiety of the formula (IX):

wherein

R V4 is hydrogen or lower alkyl;

R V6 is alkyl comprising 1 to 10 carbon atoms; and

K is an alkyl or cycloalkyl linker comprising 1 to 10 carbon atoms.

31. The binding molecule of claim 29 , wherein R V4 and R V5 are each methyl.

32. The binding molecule of claim 29 , wherein K comprises 1, 2, or 3 carbon atoms.

33. The binding molecule of claim 1 , wherein the bridging moiety is linked to Ab through a sulfide linkage.

34. The binding molecule of claim 33 , wherein B is linked to Ab through an exterior cysteine.

35. The binding molecule of claim 33 , wherein B is linked to Ab by cleaving one or more disulfide bonds in Ab.

36. The binding molecule of claim 33 , wherein B is linked to Ab without cleaving one or more disulfide bonds in Ab.

37. The binding molecule of claim 1 , wherein Ab binds to a molecule preferentially expressed on cancer cells.

38. The binding molecule of claim 1 , wherein said Ab binds to CD33.

39. The binding molecule of claim 1 , wherein said Ab binds to BR96, IgG, CD56, CD44v6, Her2/neu, Lewis, or CD30.

40. The binding molecule of claim 1 , wherein said Ab binds to Cripto.

41. The binding molecule of claim 1 , wherein said binding molecule comprises two or more Ab.

42. The binding molecule of claim 41 , wherein two or more of said Abs bind to different molecules.

43. The binding molecule of claim 2 , wherein said attachment moiety is an amino attachment moiety.

44. The binding molecule of claim 2 , wherein said attachment moiety comprises a N-succinimidyl, N-sulfosuccinimidyl, N-phthalimidyl, N-sulfophthalimidyl, 2-nitrophenyl, 4-nitrophenyl, 2,4-dinitrophenyl, 3-sulfonyl-4-nitrophenyl, or 3-carboxy-4-nitrophenyl moiety.

45. The binding molecule of claim 2 , wherein said attachment moiety is a carboxylate attachment moiety.

46. The binding molecule of claim 45 , wherein said carboxylate attachment moiety comprises an activated ester or an activated carbonyl moiety.

47. The binding molecule of claim 2 , wherein said attachment moiety is a thiol attachment moiety.

48. The binding molecule of claim 47 , wherein said thiol attachment moiety comprises an activated acyl moiety, activated alkyl group, a Michael acceptor, or an activated disulfide linkage.

49. The binding molecule of claim 2 , wherein said attachment moiety is of the formula (X):

wherein

Q is a spacer or a covalent bond.

50. The binding molecule of claim 2 , wherein said attachment moiety is of the formula (XI):

wherein

Q is a spacer or a covalent bond.

51. The binding molecule of claim 2 , wherein at least one Z is a drug moiety.

52. The binding molecule of claim 51 , wherein Z is an anticancer, anti-inflammatory, antibiotic, or anesthetic agent.

53. The binding molecule of claim 2 , wherein Z is doxorubicin, eptoside, taxane, paclitaxel, fluorouracyl, mitomycin, camptothecin, a vinca alkaloid, tubulin inhibitor, proteasome inhibitor, epothilone, cephalostatin, gemcitabine, geldanamycin, epothilone, cephalostatin, neocarzinostatin, calicheamicin, maytanisinoids, (RS)-cyclophophamide, 6-mercaptopurin, auristatin E, daunorubicin, or a derivative or analog thereof.

54. The binding molecule of claim 53 , wherein Z is a maytansinoid of the formula (IV):

wherein

R Z1 is halogen or hydrogen; and

R Z2 and R Z3 are each hydrogen or lower alkyl.

55. The binding molecule of claim 54 , wherein R Z1 is chlorine and R Z2 and R Z3 are each methyl.

56. The binding molecule of claim 2 , wherein L is cleavable.

57. The binding molecule of claim 56 , wherein L is selected such that it is cleaved extracellularly.

58. The binding molecule of claim 56 , wherein L is selected such that it is cleaved intracellularly.

59. The binding molecule of claim 2 , wherein L is of the formula (VII):

—(CR L1 R L2 ) f —NR L3 —C(═O)—(CR L4 R L5 ) g —S-D  (VII)

wherein

R L1 , R L2 , R L3 , R L4 , and R L5 are each independently alkyl, alkenyl, alkynyl, acyl, or hydrogen;

f and g are each independently selected for each occurrence from the group consisting of 0, 1, 2, 3, 4, 5, and 6; and

D is a drug attachment moiety.

60. The binding molecule of claim 59 , wherein D comprises a moiety of the formula (VIII):

wherein

R V4 and R V5 are each hydrogen or lower alkyl; and

K is an alkyl or cycloalkyl linker comprising 1 to 10 carbon atoms.

61. The binding molecule of claim 59 , wherein D comprises a moiety the formula (IX):

wherein

R V4 is hydrogen or lower alkyl;

R V6 is alkyl comprising 1 to 10 carbon atoms; and

K is an alkyl or cycloalkyl linker comprising 1 to 10 carbon atoms.

62. A method for treating a subject suffering from a disorder that would benefit from treatment with a binding molecule, comprising administering to said subject an effective amount of the binding molecule of claim 1 , such that said subject is treated.

63. The method of claim 62 , wherein said subject is suffering from cancer.

64. The method of claim 62 , wherein said subject is suffering from lymphoma.

65. The method of claim 62 , wherein said subject is suffering from an autoimmune disorder or disease.

66. The method of claim 62 , wherein said subject is suffering from an inflammatory disease or disorder.

67. A composition, comprising binding molecule of claim 1 and a pharmaceutically acceptable carrier.

68. The composition of claim 67 , wherein said pharmaceutically acceptable carrier is suitable for administration parenterally.