IP Library › Granted Patent US 11,401,337
Granted Patent B2
US 11,401,337 · App. 16/500,292 · Granted Aug 2, 2022

Modified human IgM constant regions for modulation of complement-dependent cytolysis effector function

Inventors: Ramesh Baliga (Redwood City, CA); Dean Ng (San Francisco, CA)
Assignee: IGM Biosciences, Inc.
C07K16/2887C07K16/2809C07K2317/31C07K2317/526C07K2317/622C07K2317/71C07K2317/734
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Quick Facts
Patent No.
US 11,401,337
App. No.
16/500,292
Granted
Aug 2, 2022
Kind
B2
Abstract

The disclosure provides modified human IgM heavy chain constant regions that include one or more amino acid substitutions, e.g., in the Cμ3 domain, where a modified human IgM antibody comprising the modified IgM constant region and a heavy chain variable region specific for a target antigen exhibits reduced complement-dependent cytotoxicity (CDC) of cells expressing the target antigen relative to a corresponding wild-type human IgM antibody.

Claims (41)

1. A modified human IgM constant region comprising one or more amino acid substitutions relative to a wild-type human IgM constant region, wherein at least one amino acid substitution is in the Cμ3 domain at position L310 of SEQ ID NO: 1, position P311 of SEQ ID NO: 1, position P313 of SEQ ID NO: 1, position K315 of SEQ ID NO: 1, or a combination thereof, and wherein a modified IgM antibody comprising the modified IgM constant region and a heavy chain variable region specific for a target antigen exhibits reduced complement-dependent cytotoxicity (CDC) of cells expressing the target antigen relative to a corresponding wild-type human IgM antibody identical except for the modified human IgM.

2. The modified human IgM constant region of claim 1 , wherein the amino acid substitution is at position L310 of SEQ ID NO: 1, and wherein L310 of SEQ ID NO: 1 is substituted with alanine (L310A), serine (L310S), aspartic acid (L310D) or glycine (L310G).

3. The modified human IgM constant region of claim 1 , wherein the amino acid substitution is at position P311 of SEQ ID NO: 1, and wherein P311 of SEQ ID NO: 1 is substituted with alanine (P311A), serine (P311S), or glycine (P311G).

4. The modified human IgM constant region of claim 1 , wherein the amino acid substitution is at position P313 of SEQ ID NO: 1, and wherein P313 of SEQ ID NO: 1 is substituted with alanine (P313A), serine (P313S), or glycine (P313G).

5. The modified human IgM constant region of claim 1 , wherein the amino acid substitution is at position K315 of SEQ ID NO: 1, and wherein K315 of SEQ ID NO: 1 is substituted with alanine (K315A), serine (K315S), aspartic acid (K315D), glutamine (K315Q), or glycine (K315G).

6. The modified human IgM constant region of claim 5 , wherein K315 of SEQ ID NO: 1 is substituted with aspartic acid (K315D).

7. The modified human IgM constant region of claim 1 , comprising amino acid substitutions at positions P311 and P313 of SEQ ID NO: 1, wherein P311 of SEQ ID NO: 1 is substituted with alanine (P311A), serine (P311S), or glycine (P311G), and wherein P313 of SEQ ID NO: 1 is substituted with alanine (P313A), serine (P313S), or glycine (P313G).

8. The modified human IgM constant region of claim 7 , wherein P311 of SEQ ID NO: 1 is substituted with alanine (P311A) and P313 of SEQ ID NO: 1 is substituted with serine (P313S).

9. The modified human IgM constant region of claim 1 , comprising

(a) amino acid substitutions at positions L310 and K315 of SEQ ID NO: 1, wherein L310 is substituted with alanine (L310A) or serine (L310S) and K315 is substituted with alanine (K315A) or serine (K315S);

(b) amino acid substitutions at positions L310 and P311 of SEQ ID NO: 1, wherein L310 is substituted with alanine (L310A) and P311 is substituted with alanine (P311A);

(c) amino acid substitutions at positions L310 and P313 of SEQ ID NO: 1, wherein L310 is substituted with alanine (L310A) and P313 is substituted with serine (P313S);

(d) amino acid substitutions at positions P311 and K315 of SEQ ID NO: 1, wherein P311 is substituted with alanine (P311A) and K315 is substituted with alanine (K315A); or

(e) amino acid substitutions at positions P313 and K315 of SEQ ID NO: 1, wherein P313 is substituted with serine (P313S) and K315 is substituted with alanine (K315A).

10. The modified human IgM constant region of claim 1 , comprising SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 4, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22 or SEQ ID NO: 23.

11. The modified human IgM constant region of claim 10 , wherein the maximum CDC activity achieved by a target-specific IgM antibody comprising the modified human IgM constant region in a dose-response assay is reduced by at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 100% relative to a corresponding wild-type IgM antibody identical except for the modified human IgM constant region.

12. A modified human IgM antibody comprising the modified human IgM constant region of claim 1 , and a heavy chain variable region (VH) situated amino terminal to the modified human IgM constant region, wherein the modified human IgM antibody specifically binds to a target antigen and exhibits reduced complement-dependent cytotoxicity (CDC) of cells expressing the target antigen relative to a corresponding wild-type human IgM antibody.

13. The modified human IgM antibody of claim 12 , which is a pentameric or a hexameric antibody comprising five or six bivalent IgM binding units, respectively, wherein each binding unit comprises two IgM heavy chains each comprising a VH situated amino terminal to the modified human IgM constant region, and two immunoglobulin light chains each comprising a light chain variable domain (VL) situated amino terminal to a human immunoglobulin light chain constant region.

14. The modified human IgM antibody of claim 13 , which is a pentameric antibody, and further comprises a J-chain or a functional variant thereof.

15. The modified human IgM antibody of claim 14 , wherein the J-chain or variant thereof is a modified J-chain further comprising a heterologous polypeptide, wherein the heterologous polypeptide is directly or indirectly fused to the J-chain or variant thereof.

16. The modified human IgM antibody of claim 15 , wherein the heterologous polypeptide is fused to the J-chain or variant thereof via a peptide linker to the N-terminus of the J-chain or variant thereof, the C-terminus of the J-chain or variant thereof, or to both the N-terminus and C-terminus of the J-chain or variant thereof, wherein the peptide linker comprises at least 5 amino acids, but no more than 25 amino acids.

17. The modified human IgM antibody of claim 16 , wherein the heterologous polypeptide comprises an antibody or antigen-binding fragment thereof.

18. The modified human IgM antibody of claim 17 , wherein the antigen-binding fragment is a scFv.

19. The modified human IgM antibody of claim 18 , wherein the heterologous polypeptide can specifically bind to CD3ε.

20. A polynucleotide comprising a nucleic acid sequence that encodes the modified human IgM antibody of claim 1 .

21. A composition comprising a first polynucleotide comprising a nucleic acid sequence that encodes the VH and the modified human IgM constant region of the modified human IgM antibody of claim 12 and a second polynucleotide comprising a nucleic acid sequence that encodes a light chain polypeptide comprising a human antibody light chain constant region fused to the C-terminal end of a light chain variable domain (VL).

22. The composition of claim 21 , further comprising a nucleic acid sequence that encodes a J-chain or a functional variant thereof.

23. A host cell comprising the composition of claim 21 .

24. A method of producing the modified human IgM antibody, comprising culturing the host cell of claim 23 , and recovering the modified human IgM antibody.

25. The modified human IgM constant region of claim 3 , wherein P311 of SEQ ID NO: 1 is substituted with alanine (P311A).

26. The modified human IgM constant region of claim 25 , comprising SEQ ID NO: 2.

27. The modified human IgM constant region of claim 4 , wherein P313 of SEQ ID NO: 1 is substituted with serine (P313S).

28. The modified human IgM constant region of claim 27 , comprising SEQ ID NO: 3.

29. The modified human IgM constant region of claim 8 , comprising SEQ ID NO: 4.

30. The modified human IgM constant region of claim 6 , comprising SEQ ID NO: 24.

31. The modified human IgM antibody of claim 12 , wherein the modified human IgM constant region comprises SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 4, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, or SEQ ID NO: 23.

32. The modified human IgM antibody of claim 31 , wherein the modified human IgM constant region comprises SEQ ID NO: 4.

33. The modified human IgM antibody of claim 31 , wherein the modified human IgM constant region comprises SEQ ID NO: 24.

34. The modified human IgM antibody of claim 13 , wherein the modified human IgM constant region comprises SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 4, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22 or SEQ ID NO: 23.

35. The modified human IgM antibody of claim 34 , wherein the modified human IgM constant region comprises SEQ ID NO: 4.

36. The modified human IgM antibody of claim 34 , wherein the modified human IgM constant region comprises SEQ ID NO: 24.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Oct 7, 2019
From: BALIGA, RAMESH; NG, DEAN
To: IGM BIOSCIENCES, INC.
Reel/Frame 050644/0568 →
Continuity (2)
Provisional Application 62483087 · Apr 7, 2017
Related Publication 20210147567A1 · May 20, 2021
Cited By (2)
US 12,486,336 US 12,649,791