Anti-CD79B antibodies and immunoconjugates and methods of use

1. An immunoconjugate comprising an anti-CD79b antibody comprising:

(i) an HVR-L1 sequence of KASQSVDYEGDSFLN (SEQ ID NO: 194),

(ii) an HVR-L2 sequence of AASNLES (SEQ ID NO: 195),

(iii) an HVR-L3 sequence of QQSNEDPLT (SEQ ID NO: 196),

(iv) an HVR-H1 sequence of GYTFSSYWIE (SEQ ID NO: 202),

v) an HVR-H2 sequence of GEILPGGGDTNYNEIFKG (SEQ ID NO: 203), and

(vi) an HVR-H3 sequence of TRRVPIRLDY (SEQ ID NO: 204),

wherein said anti-CD79b antibody is covalently attached to a cytotoxic agent.

2. The immunoconjugate of claim 1 , wherein the cytotoxic agent is selected from a toxin, a chemotherapeutic agent, a drug moiety, an antibiotic, a radioactive isotope and a nucleolytic enzyme.

3. The immunoconjugate of claim 2 , wherein the immunoconjugate having comprises the formula Ab-(L-D)p, wherein:

(a) Ab is an anti-CD79b antibody comprising:

(i) an HVR-L1 sequence of KASQSVDYEGDSFLN (SEQ ID NO: 194),

(ii) an HVR-L2 sequence of AASNLES (SEQ ID NO: 195),

(iii) an HVR-L3 sequence of QQSNEDPLT (SEQ ID NO: 196),

(iv) an HVR-H1 sequence of GYTFSSYWIE (SEQ ID NO: 202),

(v) an HVR-H2 sequence of GEILPGGGDTNYNEIFKG (SEQ ID NO: 203), and

(vi) an HVR-H3 sequence of TRRVPIRLDY (SEQ ID NO: 204);

(b) L is a linker;

(c) D is a drug moiety; and

(d) p ranges from about 1 to 8.

4. The immunoconjugate of claim 3 , wherein L is selected from 6-maleimidocaproyl (MC), maleimidopropanoyl (MP), valine-citrulline (val-cit), alanine-phenylalanine (ala-phe), p-aminobenzyloxycarbonyl (PAB), N-Succinimidyl 4-(2-pyridylthio) pentanoate (SPP), N-succinimidyl 4-(N-maleimidomethyl)cyclohexane-1 carboxylate N-succinimidyl 4-(N-maleimidomethyl) cyclohexane-1 carboxylate (SMCC), and N-Succinimidyl (4-iodo-acetyl)aminobenzoate N-Succinimidyl (4-iodoacetyl) aminobenzoate (SIAB).

5. The immunoconjugate of claim 3 , wherein D is selected from an auristatin and dolostatin a dolastatin.

6. The immunoconjugate of claim 3 , wherein L has comprises Formula II:

-A a -W w —Y y —  Formula II

wherein:

A is a Stretcher unit,

a is 0 or 1,

each W is independently an Amino Acid unit,

w is an integer ranging from 0 to 12,

Y is a Spacer unit, and

y is 0, 1 or 2,;

wherein p ranges from 1 to 20 8, and

wherein D is a drug moiety of Formula D E :

wherein the wavy line of D E indicates the covalent attachment site to A, W, or Y, and independently at each location:

R 2 is selected from H and C 1 -C 8 alkyl;

R 3 is selected from H, C 1 -C 8 alkyl, C 3 -C 8 carbocycle, aryl, C 1 -C 8 alkyl-aryl, C 1 -C 8 alkyl-(C 3 -C 8 carbocycle), C 3 -C 8 heterocycle and C 1 -C 8 alkyl-(C 3 -C 8 heterocycle);

R 4 is selected from H, C 1 -C 8 alkyl, C 3 -C 8 carbocycle, aryl, C 1 -C 8 alkyl-aryl, C 1 -C 8 alkyl-(C 3 -C 8 carbocycle), C 3 -C 8 heterocycle and C 1 -C 8 alkyl-(C 3 -C 8 heterocycle);

R 5 is selected from H and methyl;

or R 4 and R 5 jointly form a carbocyclic ring and have the formula —(CR a R b ) n — wherein R a and R b are independently selected from H, C 1 -C 8 alkyl and C 3 -C 8 carbocycle and n is selected from 2, 3, 4, 5 and 6;

R 6 is selected from H and C 1 -C 8 alkyl;

R 7 is selected from H, C 1 -C 8 alkyl, C 3 -C 8 carbocycle, aryl, C 1 -C 8 alkyl-aryl, C 1 -C 8 alkyl-(C 3 -C 8 carbocycle), C 3 -C 8 heterocycle and C 1 -C 8 alkyl-(C 3 -C 8 heterocycle);

each R 8 is independently selected from H, OH, C 1 -C 8 alkyl, C 3 -C 8 carbocycle and O—(C 1 -C 8 alkyl);

R 9 is selected from H and C 1 -C 8 alkyl; and

R 18 is selected from —C(R 8 ) 2 —C(R 8 ) 2 -aryl, —C(R 8 ) 2 —C(R 8 ) 2 —(C 3 -C 8 heterocycle), and —C(R 8 ) 2 —C(R 8 ) 2 —(C 3 -C 8 carbocycle).

7. The immunoconjugate of claim 6 , having An immunoconjugate comprising an anti-CD79b antibody (Ab), wherein the anti-CD79b antibody (Ab) comprises:

(i) an HVR-L1 sequence of KASQSVDYEGDSFLN (SEQ ID NO: 194),

(ii) an HVR-L2 sequence of AASNLES (SEQ ID NO: 195),

(iii) an HVR-L3 sequence of QQSNEDPLT (SEQ ID NO: 196),

(iv) an HVR-H1 sequence of GYTFSSYWIE (SEQ ID NO: 202),

(v) an HVR-H2 sequence of GEILPGGGDTNYNEIFKG (SEQ ID NO: 203), and

(vi) an HVR-H3 sequence of TRRVPIRLDY (SEQ ID NO: 204), and wherein said immunoconjugate comprises the formula:

Ab-6-maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl-monomethyl auristatin E (Ab-MC-vc-PAB-MMAE),

wherein Val is valine, and Cit is citrulline, and p ranges from 1 to about 8.

8. A pharmaceutical composition comprising the immunoconjugate of claim 3 or 6 and a pharmaceutically acceptable carrier.

9. The pharmaceutical composition of claim 8, wherein the pharmaceutical composition is lyophilized.

10. The pharmaceutical composition of claim 8, wherein the pharmaceutical composition is aqueous.

11. An immunoconjugate comprising an anti-CD79b antibody (Ab), wherein the anti-CD79b antibody (Ab) comprises the heavy chain variable domain sequence of SEQ ID NO:208 and the light chain variable domain sequence of SEQ ID NO:207, and wherein said immunoconjugate comprises the formula:

Ab-6-maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl-monomethyl auristatin E (Ab-MC-vc-PAB-MMAE),

wherein Val is valine, Cit is citrulline, and p ranges from 1 to about 8.

12. The immunoconjugate of claim 11, wherein the anti-CD79b antibody is a full-length antibody.

13. The immunoconjugate of claim 1, wherein the cytotoxic agent is the drug moiety monomethyl auristatin E (MMAE).

14. The immunoconjugate of claim 13, wherein the anti-CD79b antibody is covalently attached to MMAE via a linker, and the average number of MMAE drug moieties per anti-CD79b antibody is about 2 to about 5.

15. The immunoconjugate of claim 14, wherein the linker is connected to a sulfur atom of the anti-CD79b antibody.

16. The immunoconjugate of claim 15, wherein the linker comprises 6-maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (MC-val-cit-PAB).

17. The immunoconjugate of claim 3, wherein the linker is connected to a sulfur atom of the anti-CD79b antibody.

18. The immunoconjugate of claim 3, wherein the drug moiety is monomethyl auristatin E (MMAE).

19. The immunoconjugate of claim 18, wherein the anti-CD79b antibody is covalently attached to MMAE via a linker comprising valine-citrulline (val-cit).

20. The immunoconjugate of claim 6, wherein:

the Stretcher unit is 6-maleimidocaproyl (MC);

the Amino Acid unit is valine-citrulline (val-cit);

the drug moiety is monomethyl auristatin E (MMAE); and

the Spacer unit is p-aminobenzyloxycarbonyl (PAB);

wherein the Stretcher unit is covalently attached to a cysteine thiol of the anti-CD79b antibody; and the Spacer unit is linked to MMAE via a carbamate group.

21. The immunoconjugate of claim 3, wherein the anti-CD79b antibody comprises the heavy chain variable domain sequence of SEQ ID NO:208 and the light chain variable domain sequence of SEQ ID NO:207.

22. The immunoconjugate of claim 3, wherein the anti-CD79b antibody is a full-length antibody.

23. The immunoconjugate of claim 7, wherein the anti-CD79b antibody is a full-length antibody.

24. The immunoconjugate of claim 16, wherein the anti-CD79b antibody comprises the heavy chain variable domain sequence of SEQ ID NO:208 and the light chain variable domain sequence of SEQ ID NO:207.

25. The immunoconjugate of claim 16, wherein the anti-CD79b antibody is a full-length antibody.

26. The immunoconjugate of claim 19, wherein the anti-CD79b antibody comprises the heavy chain variable domain sequence of SEQ ID NO:208 and the light chain variable domain sequence of SEQ ID NO:207.

27. The immunoconjugate of claim 19, wherein the anti-CD79b antibody is a full-length antibody.

28. The immunoconjugate of claim 20, wherein the anti-CD79b antibody comprises the heavy chain variable domain sequence of SEQ ID NO:208 and the light chain variable domain sequence of SEQ ID NO:207.

29. The immunoconjugate of claim 20, wherein the anti-CD79b antibody is a full-length antibody.

30. A pharmaceutical composition comprising the immunoconjugate of claim 7 and a pharmaceutically acceptable carrier.

31. The pharmaceutical composition of claim 30, wherein the pharmaceutical composition is lyophilized.

32. The pharmaceutical composition of claim 30, wherein the pharmaceutical composition is aqueous.

33. A pharmaceutical composition comprising the immunoconjugate of claim 11 and a pharmaceutically acceptable carrier.

34. The pharmaceutical composition of claim 33, wherein the pharmaceutical composition is lyophilized.

35. The pharmaceutical composition of claim 33, wherein the pharmaceutical composition is aqueous.

36. A pharmaceutical composition comprising the immunoconjugate of claim 16 and a pharmaceutically acceptable carrier.

37. The pharmaceutical composition of claim 36, wherein the pharmaceutical composition is lyophilized.

38. The pharmaceutical composition of claim 36, wherein the pharmaceutical composition is aqueous.

39. A pharmaceutical composition comprising the immunoconjugate of claim 19 and a pharmaceutically acceptable carrier.

40. The pharmaceutical composition of claim 39, wherein the pharmaceutical composition is lyophilized.

41. The pharmaceutical composition of claim 39, wherein the pharmaceutical composition is aqueous.

42. A pharmaceutical composition comprising the immunoconjugate of claim 20 and a pharmaceutically acceptable carrier.

43. The pharmaceutical composition of claim 42, wherein the pharmaceutical composition is lyophilized.

44. The pharmaceutical composition of claim 42, wherein the pharmaceutical composition is aqueous.

45. An immunoconjugate comprising an anti-CD79b antibody (Ab), wherein the anti-CD79b antibody (Ab) comprises a heavy chain comprising the sequence of SEQ ID NO:308 and a light chain comprising the sequence of SEQ ID NO:307, and wherein said immunoconjugate comprises the formula:

Ab-6-maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl-monomethyl auristatin E (Ab-MC-vc-PAB-MMAE),

wherein Val is valine, Cit is citrulline, and p ranges from about 2 to about 5.

46. A pharmaceutical composition comprising the immunoconjugate of claim 45 and a pharmaceutically acceptable carrier.

47. The pharmaceutical composition of claim 46, wherein the pharmaceutical composition is lyophilized.

48. The pharmaceutical composition of claim 46, wherein the pharmaceutical composition is aqueous.

49. The immunoconjugate of claim 11, wherein p ranges from about 2 to about 5.

50. The immunoconjugate of claim 3, wherein p ranges from 1 to 8.

51. The immunoconjugate of claim 7, wherein p ranges from 1 to 8.

52. The immunoconjugate of claim 7, wherein p ranges from about 2 to about 5.

53. The immunoconjugate of claim 52, wherein p ranges from 2 to 5.

54. The immunoconjugate of claim 7, wherein p ranges from about 3 to about 4.

55. The immunoconjugate of claim 54, wherein p ranges from 3 to 4.

56. The immunoconjugate of claim 11, wherein p ranges from 1 to 8.

57. The immunoconjugate of claim 49, wherein p ranges from 2 to 5.

58. The immunoconjugate of claim 11, wherein p ranges from about 3 to about 4.

59. The immunoconjugate of claim 58, wherein p ranges from 3 to 4.

60. The immunoconjugate of claim 14, wherein the average number of MMAE drug moieties per anti-CD79b antibody is 2 to 5.

61. The immunoconjugate of claim 45, wherein p ranges from 2 to 5.

62. The immunoconjugate of claim 45, wherein p ranges from about 3 to about 4.

63. The immunoconjugate of claim 62, wherein p ranges from 3 to 4.

64. The immunoconjugate of claim 24, wherein the anti-CD79b antibody comprises a heavy chain comprising the sequence of SEQ ID NO:308 and a light chain comprising the sequence of SEQ ID NO:307.

65. The immunoconjugate of claim 24, wherein the average number of MMAE drug moieties per anti-CD79b antibody is from 2 to 5.

66. The immunoconjugate of claim 24, wherein the average number of MMAE drug moieties per anti-CD79b antibody is from about 3 to about 4.

67. The immunoconjugate of claim 66, wherein the average number of MMAE drug moieties per anti-CD79b antibody is from 3 to 4.

68. A pharmaceutical composition comprising the immunoconjugate of claim 24 and a pharmaceutically acceptable carrier.

69. The pharmaceutical composition of claim 68, wherein the pharmaceutical composition is lyophilized.

70. The pharmaceutical composition of claim 68, wherein the pharmaceutical composition is aqueous.