IP Library › Granted Patent US 9,540,438

Granted Patent B2

US 9,540,438 · App. 13/350,676 · Granted Jan 10, 2017

Aldehyde-tagged immunoglobulin polypeptides and methods of use thereof

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Quick Facts

Patent No.: US 9,540,438
App. No.: 13/350,676
Granted: Jan 10, 2017
Kind: B2

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Abstract

The present disclosure provides aldehyde-tagged immunoglobulin (Ig) polypeptides that can be converted by a formylglycine-generating enzyme to produce a 2-formylglycine (FGly)-modified Ig polypeptide. An FGly-modified Ig polypeptide can be covalently and site-specifically bound to a moiety of interest to provide an Ig conjugate. The disclosure also encompasses methods of production of such aldehyde-tagged Ig polypeptides, FGly-modified Ig polypeptides, and Ig conjugates, as well as methods of use of same.

Claims (178)

1. An immunoglobulin (Ig) polypeptide comprising a sulfatase motif comprising an amino acid sequence LCTPSR (SEQ ID NO:17), the Ig polypeptide comprising:

a) an IgG 1 heavy chain constant region comprising an amino acid sequence selected from the group consisting of:

ASTKGLCTPSRV (SEQ ID NO: 231),

LCTPSRSKSTSGGT (SEQ ID NO: 234),

NSGALCTPSRGVHTFPAVLQSSGL (SEQ ID NO:235),

EPKSCDKTHTCPPCPLCTPSRELLGG (SEQ ID NO: 236),

NLCTPSRAP (SEQ ID NO: 206),

SKAKGLCTPSRE (SEQ ID NO: 237), and

DVSHEDLCTPSREV (SEQ ID NO: 250);

b) an IgG 4 heavy chain constant region comprising an amino acid sequence selected from the group consisting of:

NSGALCTPSRGVHTFPAVLQSSGL (SEQ ID NO:235), and

SKAKGLCTPSRE (SEQ ID NO:237);

c) an Ig kappa light chain constant region comprising the amino acid sequence:

DNALCTPSRQSGN (SEQ ID NO: 220).

2. The Ig polypeptide of claim 1 , wherein the Ig polypeptide comprises:

a) an IgG 1 heavy chain constant region comprising an amino acid sequence selected from the group consisting of

ASTKGLCTPSRV (SEQ ID NO: 231),

LCTPSRSKSTSGGT (SEQ ID NO: 234),

NSGALCTPSRGVHTFPAVLQSSGL (SEQ ID NO: 235),

EPKSCDKTHTCPPCPLCTPSRELLGG (SEQ ID NO: 236),

NLCTPSRAP (SEQ ID NO: 206),

SKAKGLCTPSRE (SEQ ID NO: 237), and

DVSHEDLCTPSREV (SEQ ID NO: 250); or

b) an IgG 4 heavy chain constant region comprising an amino acid sequence selected from the group consisting of

NSGALCTPSRGVHTFPAVLQSSGL (SEQ ID NO:235), and

SKAKGLCTPSRE (SEQ ID NO:237).

3. The Ig polypeptide of claim 1 , wherein the Ig polypeptide comprises an Ig kappa light chain constant region comprising the amino acid sequence DNALCTPSRQSGN (SEQ ID NO: 220).

4. An antibody comprising the Ig polypeptide of claim 2 .

5. An antibody comprising the Ig polypeptide of claim 3 .

6. An immunoglobulin (Ig) polypeptide comprising a 2-formylglycine (FGly) moiety, comprising:

a) an IgG 1 heavy chain constant region comprising an amino acid sequence selected from the group consisting of:

ASTKGL(FGly)TPSRV (SEQ ID NO: 232),

L(FGly)TPSRSKSTSGGT (SEQ ID NO: 238),

NSGAL(FGly)TPSRGVHTFPAVLQSSGL (SEQ ID NO: 239),

EPKSCDKTHTCPPCPL(FGly)TPSRELLGG (SEQ ID NO: 240),

NL(FGly)TPSRAP (SEQ ID NO: 241),

SKAKGL(FGly)TPSRE (SEQ ID NO: 242), and

DVSHEDL(FGly)TPSREV (SEQ ID NO:251);

b) an IgG 4 heavy chain constant region comprising an amino acid sequence selected from the group consisting of:

NSGAL(FGly)TPSRGVHTFPAVLQSSGL (SEQ ID NO:239), and

SKAKGL(CGly)TPSRE (SEQ ID NO:242);

c) an Ig light chain constant region comprising the amino acid sequence:

DNAL(FGly)TPSRQSGN (SEQ ID NO: 248).

7. The immunoglobulin (Ig) polypeptide of claim 6 , wherein the Ig polypeptide comprises:

a) an IgG 1 heavy chain constant region comprising an amino acid sequence selected from the group consisting of:

ASTKGL(FGly)TPSRV (SEQ ID NO: 232),

L(FGly)TPSRSKSTSGGT (SEQ ID NO: 238),

NSGAL(FGly)TPSRGVHTFPAVLQSSGL (SEQ ID NO: 239),

EPKSCDKTHTCPPCPL(FGly)TPSRELLGG (SEQ ID NO: 240),

NL(FGly)TPSRAP (SEQ ID NO: 241),

SKAKGL(FGly)TPSRE (SEQ ID NO: 242), and

DVSHEDL(FGly)TPSREV (SEQ ID NO:251); or

b) an IgG 4 heavy chain constant region comprising an amino acid sequence selected from the group consisting of

NSGAL(FGly)TPSRGVHTFPAVLQSSGL (SEQ ID NO:239), and

SKAKGL(FGly)TPSRE (SEQ ID NO:242).

8. The immunoglobulin (Ig) polypeptide of claim 6 , wherein the Ig polypeptide comprises an Ig kappa light chain constant region comprising the amino acid sequence DNAL(FGly)TPSRQSGN (SEQ ID NO: 248).

9. An antibody comprising a formylglycine (FGly) moiety, wherein the antibody comprises the Ig polypeptide of claim 7 .

10. An antibody comprising a formylglycine (FGly) moiety, wherein the antibody comprises the Ig polypeptide of claim 8 .

11. An immunoglobulin (Ig) conjugate comprising:

an Ig polypeptide and a covalently bound moiety,

wherein the Ig polypeptide comprises:

a) an IgG 1 heavy chain constant region comprising an amino acid sequence selected from the group consisting of:

ASTKGL(FGly′)TPSRV (SEQ ID NO: 233),

L(FGly′)TPSRSKSTSGGT (SEQ ID NO: 243),

NSGAL(FGly′)TPSRGVHTFPAVLQSSGL (SEQ ID NO: 244),

EPKSCDKTHTCPPCPL(FGly′)TPSRELLGG (SEQ ID NO: 245),

NL(FGly′)TPSRAP (SEQ ID NO: 246),

SKAKGL(FGly′)TPSRE (SEQ ID NO: 247), and

DVSHEDL(FGly′)TPSREV (SEQ ID NO:252);

b) an IgG 4 heavy chain constant region comprising an amino acid sequence selected from the group consisting of:

NSGAL(FGly′)TPSRGVHTFPAVLQSSGL (SEQ ID NO:244), and

SKAKGL(FGly′)TPSRE (SEQ ID NO:247);

c) an Ig light chain constant region comprising DNAL(FGly′)TPSRQSGN (SEQ ID NO: 249);

and wherein FGly′ is of the formula:

wherein J 1 is the covalently bound moiety;

each L 1 is independently selected from alkylene, substituted alkylene, alkenylene, substituted alkenylene, alkynylene, arylene, substituted arylene, cycloalkylene, substituted cycloalkylene, heteroarylene, substituted heteroarylene, heterocyclene, substituted heterocyclene, acyl, amido, acyloxy, urethanylene, thioester, sulfonyl, sulfonamide, sulfonyl ester, O, S, NH, and substituted amine; and

n is a number selected from zero to 40.

12. The immunoglobulin (Ig) conjugate of claim 11 , wherein the Ig polypeptide comprises:

a) an IgG 1 heavy chain constant region comprising an amino acid sequence selected from the group consisting of:

ASTKGL(FGly′)TPSRV (SEQ ID NO: 233),

L(FGly′)TPSRSKSTSGGT (SEQ ID NO: 243),

NSGAL(FGly′)TPSRGVHTFPAVLQSSGL (SEQ ID NO: 244),

EPKSCDKTHTCPPCPL(FGly′)TPSRELLGG (SEQ ID NO: 245),

NL(FGly′)TPSRAP (SEQ ID NO: 246),

SKAKGL(FGly′)TPSRE (SEQ ID NO: 247), and

DVSHEDL(FGly′)TPSREV (SEQ ID NO:252); or

b) an IgG 4 heavy chain constant region comprising an amino acid sequence selected from the group consisting of:

NSGAL(FGly′)TPSRGVHTFPAVLQSSGL (SEQ ID NO:244), and

SKAKGL(FGly′)TPSRE (SEQ ID NO:247).

13. The immunoglobulin (Ig) conjugate of claim 11 , wherein the Ig polypeptide comprises an Ig kappa light chain constant region comprising DNAL(FGly′)TPSRQSGN (SEQ ID NO: 249).

14. The immunoglobulin (Ig) conjugate of claim 11 , wherein the Ig polypeptide comprises an IgG 1 heavy chain constant region comprising the amino acid sequence ASTKGL(FGly′)TPSRV (SEQ ID NO: 233).

15. The immunoglobulin (Ig) conjugate of claim 11 , wherein the Ig polypeptide comprises an IgG 1 heavy chain constant region comprising the amino acid sequence L(FGly′)TPSRSKSTSGGT (SEQ ID NO: 243).

16. The immunoglobulin (Ig) conjugate of claim 11 , wherein the Ig polypeptide comprises an IgG 1 or IgG 4 heavy chain constant region comprising the amino acid sequence NSGAL(FGly′)TPSRGVHTFPAVLQSSGL (SEQ ID NO: 244).

17. The immunoglobulin (Ig) conjugate of claim 11 , wherein the Ig polypeptide comprises an IgG 1 heavy chain constant region comprising the amino acid sequence EPKSCDKTHTCPPCPL(FGly′)TPSRELLGG (SEQ ID NO: 245).

18. The immunoglobulin (Ig) conjugate of claim 11 , wherein the Ig polypeptide comprises an IgG 1 heavy chain constant region comprising the amino acid sequence NL(FGly′)TPSRAP (SEQ ID NO: 246).

19. The immunoglobulin (Ig) conjugate of claim 11 , wherein the Ig polypeptide comprises an IgG 1 or IgG 4 heavy chain constant region comprising the amino acid sequence SKAKGL(FGly′)TPSRE (SEQ ID NO: 247).

20. The immunoglobulin (Ig) conjugate of claim 11 , wherein the Ig polypeptide comprises an IgG 1 heavy chain constant region comprising the amino acid sequence DVSHEDL(FGly′)TPSREV (SEQ ID NO:252).

21. The immunoglobulin (Ig) conjugate of claim 11 , wherein the Ig polypeptide comprises an Ig light chain constant region comprising the amino acid sequence DNAL(FGly′)TPSRQSGN (SEQ ID NO: 249).

22. The Ig conjugate of claim 11 , wherein J 1 is selected from a drug, a detectable label, a water-soluble polymer, and a synthetic peptide.

23. The Ig conjugate of claim 11 , wherein J 1 is a small molecule drug.

24. The Ig conjugate of claim 23 , wherein the small molecule drug is a cancer chemotherapeutic agent.

25. The Ig conjugate of claim 24 , wherein the Ig polypeptide comprises:

a) an IgG 1 heavy chain constant region comprising an amino acid sequence selected from the group consisting of:

ASTKGL(FGly′)TPSRV (SEQ ID NO: 233),

L(FGly′)TPSRSKSTSGGT (SEQ ID NO: 243),

NSGAL(FGly′)TPSRGVHTFPAVLQSSGL (SEQ ID NO: 244),

EPKSCDKTHTCPPCPL(FGly′)TPSRELLGG (SEQ ID NO: 245),

NL(FGly′)TPSRAP (SEQ ID NO: 246),

SKAKGL(FGly′)TPSRE (SEQ ID NO: 247), and

DVSHEDL(FGly′)TPSREV (SEQ ID NO:252); or

b) an IgG 4 heavy chain constant region comprising an amino acid sequence selected from the group consisting of:

NSGAL(FGly′)TPSRGVHTFPAVLQSSGL (SEQ ID NO:235), and

SKAKGL(FGly′)TPSRE (SEQ ID NO:237).

26. The Ig conjugate of claim 24 , wherein the Ig polypeptide comprises an Ig kappa light chain constant region comprising the amino acid sequence DNAL(FGly′)TPSRQSGN (SEQ ID NO: 249).

27. The Ig conjugate of claim 11 , wherein J 1 is a water-soluble polymer.

28. The Ig conjugate of claim 27 , wherein the water-soluble polymer is poly(ethylene glycol).

29. The Ig conjugate of claim 11 , wherein J 1 is a detectable label.

30. The Ig conjugate of claim 29 , wherein the detectable label is an imaging agent.

31. An antibody conjugate comprising an immunoglobulin (Ig) conjugate of claim 12 .

32. An antibody conjugate comprising an immunoglobulin (Ig) conjugate of claim 13 .

33. The antibody conjugate of claim 31 , wherein the antibody specifically binds a tumor antigen on a cancer cell.

34. The antibody conjugate of claim 33 , wherein the J 1 moiety is a cytotoxic agent.

35. The antibody conjugate of claim 32 , wherein the antibody specifically binds a tumor antigen on a cancer cell.

36. The Ig conjugate of claim 35 , wherein the J 1 moiety is a cytotoxic agent.

37. The antibody conjugate of claim 31 , wherein the antibody conjugate specifically binds an antigen on a cell infected by a virus.

38. The antibody conjugate of claim 37 , wherein the antigen is encoded by the virus.

39. The antibody conjugate of claim 31 , wherein the J 1 moiety is a viral fusion inhibitor.

40. A formulation comprising:

a) an Ig conjugate of claim 12 ; and

b) a pharmaceutically acceptable excipient.

41. A formulation comprising:

a) an Ig conjugate of claim 13 ; and

b) a pharmaceutically acceptable excipient.

42. The immunoglobulin (Ig) polypeptide of claim 1 , comprising:

a) an IgG 1 heavy chain constant region comprising an amino acid sequence selected from the group consisting of:

SEQ ID NO:253,

SEQ ID NO:256,

SEQ ID NO:259,

SEQ ID NO:262,

SEQ ID NO:265,

SEQ ID NO:268, and

SEQ ID NO:271; or

b) an IgG 4 heavy chain constant region comprising an amino acid sequence selected from the group consisting of:

SEQ ID NO:277, and

SEQ ID NO:280; or

c) an Ig kappa light chain constant region comprising an amino acid sequence set forth in SEQ ID NO:274.

43. The Ig polypeptide of claim 6 , comprising:

a) an IgG 1 heavy chain constant region comprising an amino acid sequence selected from the group consisting of:

SEQ ID NO:254,

SEQ ID NO:257,

SEQ ID NO:260,

SEQ ID NO:263,

SEQ ID NO:266,

SEQ ID NO:269, and

SEQ ID NO:272; or

b) an IgG 4 heavy chain constant region comprising an amino acid sequence selected from the group consisting of:

SEQ ID NO:278, and

SEQ ID NO:281; or

c) an Ig kappa light chain constant region comprising an amino acid sequence set forth in SEQ ID NO:275.

44. The Ig conjugate of claim 11 , wherein the Ig polypeptide comprises:

a) an IgG 1 heavy chain constant region comprising an amino acid sequence selected from the group consisting of:

SEQ ID NO:255,

SEQ ID NO:258,

SEQ ID NO:261,

SEQ ID NO:264,

SEQ ID NO:269,

SEQ ID NO:270, and

SEQ ID NO:273; or

b) an IgG 4 heavy chain constant region comprising an amino acid sequence selected from the group consisting of:

SEQ ID NO:279, and

SEQ ID NO:282; or

c) an Ig kappa light chain constant region comprising an amino acid sequence set forth in SEQ ID NO:276.

Assignments (5)

RELEASE OF SECURITY INTEREST IN PATENT COLLATERAL (034414/0616)) Recorded Dec 19, 2024

From: JPMORGAN CHASE BANK, N.A.

To: REDWOOD BIOSCIENCE, INC.

Reel/Frame 069751/0128 →

SECURITY INTEREST Recorded Dec 19, 2024

From: CATALENT CTS (KANSAS CITY), LLC; REDWOOD BIOSCIENCE, INC.; R.P. SCHERER TECHNOLOGIES, LLC; CATALENT WELLNESS, LLC; CATALENT PHARMA SOLUTIONS, INC.; CATALENT WELLNESS NEW JERSEY, LLC; CATALENT MARYLAND, INC.; CATALENT GREENVILLE, INC.; CATALENT MICRON TECHNOLOGIES, INC.; CATALENT SAN DIEGO, INC.; CATALENT WELLNESS VIRGINIA, LLC; CATALENT USA PACKAGING, LLC; CATALENT PHARMA SOLUTIONS, LLC

To: ARES CAPITAL CORPORATION, AS COLLATERAL AGENT

Reel/Frame 069743/0458 →

NOTICE OF SUCCESSION OF AGENCY Recorded May 17, 2019

From: MORGAN STANLEY SENIOR FUNDING, INC.

To: JPMORGAN CHASE BANK, N.A.

Reel/Frame 049217/0981 →

ADDITIONAL RESTRICTED SUBSIDIARY INTELLECTUAL PROPERTY SECURITY AGREEMENT SUPPLEMENT Recorded Nov 21, 2014

From: REDWOOD BIOSCIENCE, INC.

To: MORGAN STANLEY SENIOR FUNDING, INC.

Reel/Frame 034414/0616 →

ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Feb 16, 2012

From: BARFIELD, ROBYN M.; BREIDENBACH, MARK ALAN; DEHART, GREGORY W.; RABUKA, DAVID

To: REDWOOD BIOSCIENCE, INC.

Reel/Frame 027719/0831 →