IP Library › Granted Patent US 10,035,853

Granted Patent B2

US 10,035,853 · App. 15/056,490 · Granted Jul 31, 2018

Site-specific antibody conjugation methods and compositions

View Patent ↗

Loading inventors, assignments & file history…

Monitor This Case

Get email alerts when status or documents change.

Order Certified Copies

Most orders are placed with the USPTO same day — all within 24 business hours.

Order via The Patent Place →

Pre-filled with this patent's details

Quick Facts

Patent No.: US 10,035,853
App. No.: 15/056,490
Granted: Jul 31, 2018
Kind: B2

External Links

USPTO Patent Center↗ Full Patent PDF↗

Abstract

Provided are novel antibody drug conjugates (ADCs), and methods of using such ADCs to treat proliferative disorders.

Claims (102)

1. An engineered IgG1 monoclonal antibody comprising one or more unpaired cysteine residues, wherein the engineered antibody binds SEZ6;

wherein the engineered antibody comprises (i) a cysteine residue at heavy chain position 220 and a deletion of a cysteine residue at light chain position 214, (ii) a cysteine residue at heavy chain position 220 and substitution of a cysteine residue at light chain position 214, (iii) a cysteine residue at light chain position 214 and a deletion of a cysteine residue at heavy chain position 220, or (iv) a cysteine residue at light chain position 214 and a substitution of a cysteine residue at heavy chain position 220;

wherein the engineered antibody comprises native cysteine residues at heavy chain positions 226 and 229;

and wherein the engineered antibody comprises (a) three CDRs of a light chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 190 or 192; and three CDRs of a heavy chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 191; or (b) three CDRs of a light chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 172; and three CDRs of a heavy chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 173.

2. The engineered antibody of claim 1 wherein the engineered antibody comprises a humanized antibody or a CDR grafted antibody.

3. The engineered antibody of claim 1 wherein the engineered antibody comprises two light chains, two heavy chains and two unpaired cysteine residues.

4. The engineered antibody of claim 3 wherein

each of the two light chains comprises an unpaired cysteine residue at position 214.

5. The engineered antibody of claim 3 wherein

each of the two heavy chains comprises an unpaired cysteine residue at position C220.

6. An antibody drug conjugate, comprising an engineered IgG1 monoclonal antibody and a cytotoxic agent, wherein the engineered antibody binds SEZ6;

wherein the engineered antibody comprises native cysteine residues at heavy chain positions 226 and 229;

wherein the engineered antibody comprises: (a) three CDRs of a light chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 190 or 192, and three CDRs of a heavy chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 191; or (b) three CDRs of a light chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 172, and three CDRs of a heavy chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 173; and

wherein the antibody is conjugated to the cytotoxic agent via the cysteine residue at light chain position 214 or the cysteine residue at heavy chain position 220.

7. The antibody drug conjugate of claim 6 wherein the engineered antibody comprises a humanized antibody or a CDR grafted antibody.

8. The antibody drug conjugate of claim 6 wherein the engineered antibody comprises two light chains, two heavy chains and two unpaired cysteine residues.

9. The antibody drug conjugate of claim 6 , wherein the engineered antibody comprises two light chains and two heavy chains, wherein

each of the two light chains comprises a cysteine residue at position 214.

10. The antibody drug conjugate of claim 6 , wherein the engineered antibody comprises two light chains and two heavy chains, wherein

each of the two heavy chains comprises a cysteine residue at position C220.

11. A pharmaceutical composition comprising the engineered antibody of claim 1 and a pharmaceutically acceptable carrier.

12. A pharmaceutical composition comprising the antibody drug conjugate of claim 6 and a pharmaceutically acceptable carrier.

13. The engineered antibody of claim 1 , comprising

(a) residues 24-34 of SEQ ID NO: 190 for CDR-L1, residues 50-56 of SEQ ID NO: 190 for CDR-L2, residues 89-97 of SEQ ID NO: 190 for CDR-L3, residues 26-32 of SEQ ID NO: 191 for CDR-H1, residues 52-56 of SEQ ID NO: 191 for CDR-H2, and residues 95-102 of SEQ ID NO: 191 for CDR-H3, wherein the residues are numbered according to Chothia; or

(b) residues 24-34 of SEQ ID NO: 192 for CDR-L1, residues 50-56 of SEQ ID NO: 192 for CDR-L2, residues 89-97 of SEQ ID NO: 192 for CDR-L3, residues 26-32 of SEQ ID NO: 191 for CDR-H1, residues 52-56 of SEQ ID NO: 191 for CDR-H2, and residues 95-102 of SEQ ID NO: 191 for CDR-H3, wherein the residues are numbered according to Chothia.

14. The engineered antibody of claim 1 , comprising

(a) residues 30-36 of SEQ ID NO: 190 for CDR-L1, residues 46-55 of SEQ ID NO: 190 for CDR-L2, residues 89-96 of SEQ ID NO: 190 for CDR-L3, residues 30-35 of SEQ ID NO: 191 for CDR-H1, residues 47-58 of SEQ ID NO: 191 for CDR-H2, and residues 93-101 of SEQ ID NO: 191 for CDR-H3, wherein the residues are numbered according to MacCallum; or

(b) residues 30-36 of SEQ ID NO: 192 for CDR-L1, residues 46-55 of SEQ ID NO: 192 for CDR-L2, residues 89-96 of SEQ ID NO: 192 for CDR-L3, residues 30-35 of SEQ ID NO: 191 for CDR-H1, residues 47-58 of SEQ ID NO: 191 for CDR-H2, and residues 93-101 of SEQ ID NO: 191 for CDR-H3, wherein the residues are numbered according to MacCallum.

15. The engineered antibody of claim 1 , comprising

(a) residues 24-34 of SEQ ID NO: 190 for CDR-L1, residues 50-56 of SEQ ID NO: 190 for CDR-L2, residues 89-97 of SEQ ID NO: 190 for CDR-L3, residues 31-35 of SEQ ID NO: 191 for CDR-H1, residues 50-65 of SEQ ID NO: 191 for CDR-H2, and residues 95-102 of SEQ ID NO: 191 for CDR-H3, wherein the residues are numbered according to Kabat; or

(b) residues 24-34 of SEQ ID NO: 192 for CDR-L1, residues 50-56 of SEQ ID NO: 192 for CDR-L2, residues 89-97 of SEQ ID NO: 192 for CDR-L3, residues 31-35 of SEQ ID NO: 191 for CDR-H1, residues 50-65 of SEQ ID NO: 191 for CDR-H2, and residues 95-102 of SEQ ID NO: 191 for CDR-H3, wherein the residues are numbered according to Kabat.

16. The engineered antibody of claim 1 , wherein the engineered antibody comprises

(a) a light chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 403 and a heavy chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 500;

(b) a light chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 403 and a heavy chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 501;

(c) a light chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 502 and a heavy chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 404; or

(d) a light chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 503 and a heavy chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 404.

17. The engineered antibody of claim 1 , wherein the engineered antibody comprises

(a) a light chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 504 and a heavy chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 500;

(b) a light chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 504 and a heavy chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 501;

(c) a light chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 505 and a heavy chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 404; or

(d) a light chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 506 and a heavy chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 404.

18. The engineered antibody of claim 1 comprising:

(a) a light chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 190 and a heavy chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 191;

(b) a light chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 192 and a heavy chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 191; or

(c) a light chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 172 and a heavy chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 173.

19. The engineered antibody of claim 1 comprising:

(a) a light chain comprising an amino acid sequence set forth as SEQ ID NO: 513 and a heavy chain comprising an amino acid sequence set forth as SEQ ID NO: 515;

(b) a light chain comprising an amino acid sequence set forth as SEQ ID NO: 513 and a heavy chain comprising an amino acid sequence set forth as SEQ ID NO: 516;

(c) a light chain comprising an amino acid sequence set forth as SEQ ID NO: 517 and a heavy chain comprising an amino acid sequence set forth as SEQ ID NO: 514; or

(d) a light chain comprising an amino acid sequence set forth as SEQ ID NO: 518 and a heavy chain comprising an amino acid sequence set forth as SEQ ID NO: 514.

20. The engineered antibody of claim 1 , wherein the engineered antibody is conjugated to a cytotoxic agent via the cysteine residue at light chain position 214 or the cysteine residue at heavy chain position 220.

21. The engineered antibody of claim 20 , wherein the cytotoxic agent is an auristatin, a maytansinoid, a calicheamicin or a radioisotope.

22. The antibody drug conjugate of claim 6 , wherein the engineered antibody comprises:

23. The antibody drug conjugate of claim 6 , wherein the engineered antibody comprises:

24. The antibody drug conjugate of claim 6 , wherein the engineered antibody comprises:

25. The antibody drug conjugate of claim 6 , wherein the engineered antibody comprises:

(a) a light chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 403 and a heavy chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 500;

(b) a light chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 403 and a heavy chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 501;

(c) a light chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 502 and a heavy chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 404; or

(d) a light chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 503 and a heavy chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 404.

26. The antibody drug conjugate of claim 6 , wherein the engineered antibody comprises:

(a) a light chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 504 and a heavy chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 500;

(b) a light chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 504 and a heavy chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 501;

(c) a light chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 505 and a heavy chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 404; or

(d) a light chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 506 and a heavy chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 404.

27. The antibody drug conjugate of claim 6 , wherein the engineered antibody comprises:

(a) a light chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 190 and a heavy chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 191;

(b) a light chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 192 and a heavy chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 191; or

(c) a light chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 172 and a heavy chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 173.

28. The antibody drug conjugate of claim 6 , wherein the engineered antibody comprises:

(a) a light chain comprising an amino acid sequence set forth as SEQ ID NO: 513 and a heavy chain comprising an amino acid sequence set forth as SEQ ID NO: 515;

(b) a light chain comprising an amino acid sequence set forth as SEQ ID NO: 513 and a heavy chain comprising an amino acid sequence set forth as SEQ ID NO: 516;

(c) a light chain comprising an amino acid sequence set forth as SEQ ID NO: 517 and a heavy chain comprising an amino acid sequence set forth as SEQ ID NO: 514; or

(d) a light chain comprising an amino acid sequence set forth as SEQ ID NO: 518 and a heavy chain comprising an amino acid sequence set forth as SEQ ID NO: 514.

29. The antibody drug conjugate of claim 6 , wherein the cytotoxic agent is an auristatin, a maytansinoid, a calicheamicin or a radioisotope.

30. The antibody drug conjugate of claim 6 , wherein the antibody drug conjugate has a drug loading of 2.

31. The pharmaceutical composition of claim 12 comprising an average drug to antibody ratio (DAR) of 2+/−0.4.

32. The pharmaceutical composition of claim 12 , wherein the predominant antibody drug conjugate species is present at a concentration of greater than 70%.

33. An engineered IgG1 monoclonal antibody comprising one or more unpaired cysteine residues, wherein the engineered antibody binds SEZ6;

wherein the engineered antibody comprises (i) a cysteine residue at light chain position 214 and a deletion of a cysteine residue at heavy chain position 220 or (ii) a cysteine residue at light chain position 214 and a substitution of a cysteine residue at heavy chain position 220;

wherein the engineered antibody comprises native cysteine residues at heavy chain positions 226 and 229;

and wherein the engineered antibody comprises three CDRs of a light chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 190 and three CDRs of a heavy chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 191.

34. The engineered antibody of claim 33 , wherein the engineered antibody comprises a light chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 403 and a heavy chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 500.

35. The engineered antibody of claim 34 , wherein the engineered antibody comprises residues 24-34 of SEQ ID NO: 190 for CDR-L1, residues 50-56 of SEQ ID NO: 190 for CDR-L2, residues 89-97 of SEQ ID NO: 190 for CDR-L3, residues 26-32 of SEQ ID NO: 191 for CDR-H1, residues 52-56 of SEQ ID NO: 191 for CDR-H2, and residues 95-102 of SEQ ID NO: 191 for CDR-H3, wherein the residues are numbered according to Chothia.

36. The engineered antibody of claim 34 , wherein the engineered antibody comprises residues 30-36 of SEQ ID NO: 190 for CDR-L1, residues 46-55 of SEQ ID NO: 190 for CDR-L2, residues 89-96 of SEQ ID NO: 190 for CDR-L3, residues 30-35 of SEQ ID NO: 191 for CDR-H1, residues 47-58 of SEQ ID NO: 191 for CDR-H2, and residues 93-101 of SEQ ID NO: 191 for CDR-H3, wherein the residues are numbered according to MacCallum.

37. The engineered antibody of claim 34 , wherein the engineered antibody comprises residues 24-34 of SEQ ID NO: 190 for CDR-L1, residues 50-56 of SEQ ID NO: 190 for CDR-L2, residues 89-97 of SEQ ID NO: 190 for CDR-L3, residues 31-35 of SEQ ID NO: 191 for CDR-H1, residues 50-65 of SEQ ID NO: 191 for CDR-H2, and residues 95-102 of SEQ ID NO: 191 for CDR-H3, wherein the residues are numbered according to Kabat.

38. An antibody drug conjugate, comprising an engineered IgG1 monoclonal antibody and a cytotoxic agent, wherein the engineered antibody binds SEZ6;

wherein the engineered antibody comprises (i) a cysteine residue at light chain position 214 and a deletion of a cysteine residue at heavy chain position 220, or (ii) a cysteine residue at light chain position 214 and a substitution of a cysteine residue at heavy chain position 220;

wherein the engineered antibody comprises native cysteine residues at heavy chain positions 226 and 229;

wherein the engineered antibody comprises three CDRs of a light chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 190, and three CDRs of a heavy chain variable region comprising an amino acid sequence set forth as SEQ ID NO: 191; and

wherein the antibody is conjugated to the cytotoxic agent via the cysteine residue at light chain position 214.

39. The antibody drug conjugate of claim 38 , wherein the engineered antibody comprises a light chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 403 and a heavy chain constant region comprising an amino acid sequence set forth as SEQ ID NO: 500.

40. The antibody drug conjugate of claim 39 , wherein the engineered antibody comprises residues 24-34 of SEQ ID NO: 190 for CDR-L1, residues 50-56 of SEQ ID NO: 190 for CDR-L2, residues 89-97 of SEQ ID NO: 190 for CDR-L3, residues 26-32 of SEQ ID NO: 191 for CDR-H1, residues 52-56 of SEQ ID NO: 191 for CDR-H2, and residues 95-102 of SEQ ID NO: 191 for CDR-H3, wherein the residues are numbered according to Chothia.

41. The antibody drug conjugate of claim 39 , wherein the engineered antibody comprises residues 30-36 of SEQ ID NO: 190 for CDR-L1, residues 46-55 of SEQ ID NO: 190 for CDR-L2, residues 89-96 of SEQ ID NO: 190 for CDR-L3, residues 30-35 of SEQ ID NO: 191 for CDR-H1, residues 47-58 of SEQ ID NO: 191 for CDR-H2, and residues 93-101 of SEQ ID NO: 191 for CDR-H3, wherein the residues are numbered according to MacCallum.

42. The antibody drug conjugate of claim 39 , wherein the engineered antibody comprises residues 24-34 of SEQ ID NO: 190 for CDR-L1, residues 50-56 of SEQ ID NO: 190 for CDR-L2, residues 89-97 of SEQ ID NO: 190 for CDR-L3, residues 31-35 of SEQ ID NO: 191 for CDR-H1, residues 50-65 of SEQ ID NO: 191 for CDR-H2, and residues 95-102 of SEQ ID NO: 191 for CDR-H3, wherein the residues are numbered according to Kabat.

Assignments (4)

NUNC PRO TUNC ASSIGNMENT Recorded Oct 25, 2018

From: ARATHOON, WILLIAM ROBERT; PADAWER, ISHAI; CANO, LUIS ANTONIO; SISODIYA, VIKRAM NATWARSINHJI; MANI, KARTHIK NARAYAN; LIU, DAVID

To: ABBVIE STEMCENTRX LLC

Reel/Frame 047313/0386 →

MERGER Recorded Aug 7, 2016

From: STEMCENTRX, INC.

To: ABBVIE STEMCENTRX LLC

Reel/Frame 039601/0189 →

CHANGE OF NAME Recorded Jul 15, 2016

From: STEM CENTRX, INC.

To: STEMCENTRX, INC.