Fold-back diabody diphtheria toxin immunotoxin and methods of use

Patent metadata

Field	Value
Publication number	US-9364557-B2
Application number	US-67162908-A
Country	US
Kind code	B2
Filing date	Aug 1, 2008
Priority date	Aug 1, 2007
Publication date	Jun 14, 2016
Grant date	Jun 14, 2016

Abstract

Provided are methods and compositions related to diphtheria toxin diabody immunotoxins.

First claim

What is claimed is:

1. An immunotoxin fusion protein produced in Pichia pastoris , comprising a mutant diphtheria toxin moiety and a targeting moiety, wherein the targeting moiety is a single chain variable region bivalent diabody comprising a first scFv and a second scFv, wherein said first scFv and said second scFv share the same antigen specificity, and wherein the diabody comprises an anti-PSMA V L V H region.

2. The immunotoxin of claim 1 , wherein the diabody is a fold-back diabody.

3. The immunotoxin of claim 1 , wherein the diabody anti-PSMA V L V H region comprises the J591 V L V H region.

4. The immunotoxin of claim 1 , wherein the toxin moiety is a truncation mutant.

5. The immunotoxin of claim 4 , wherein the truncation mutant comprises a deletion of 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, or 150 C-terminal amino acids of the full length diphtheria toxin.

6. The immunotoxin of claim 4 , wherein the immunotoxin is A-dmDT390scfbDb(J591).

7. The immunotoxin of claim 6 , wherein the immunotoxin is A-dmDT390scfbDb(J591) where the furin site amino acid sequence cleavage site RVRRSV (SEQ ID NO: 24) has been replaced with the MMP cleavage site GPLGMLSQ (SEQ ID NO: 25).

8. The immunotoxin of claim 6 , wherein the immunotoxin is A-dmDT390scfbDb(J591) where the furin cleavage site amino acid sequence RVRRSV (SEQ ID NO: 24) has been replaced with the PSA cleavage site HSSKLQ (SEQ ID NO: 26).

9. An immunotoxin fusion protein comprising a diphtheria toxin moiety and a targeting moiety, wherein the targeting moiety is a diabody, wherein said diabody comprises a first scFv and a second scFv, wherein said first scFv and said second scFv share the same antigen specificity, wherein the sequence from the amino terminus from left to right is toxin moiety V L1 , L 2 , V H1 , L 3 , V L2 , L 4 , V H2 , wherein the toxin moiety comprises a truncation mutation L 2 and L 4 are (G 4 S) linkers, L 3 is a (G 4 S) 3 linker, and V L and V H are the variable light and heavy domains, wherein the V L1 and V H1 comprise the V L V H region of an anti-PSMA antibody.

10. The immunotoxin of claim 9 , wherein the diabody is a fold-back diabody.

11. The immunotoxin of claim 9 , wherein the anti-PSMA antibody is J591.

12. The immunotoxin of claim 9 , wherein V L1 and V H1 are the same as V L2 and V H2 .

13. The immunotoxin of claim 9 , wherein the toxin moiety is a truncation mutant.

14. The immunotoxin of claim 13 , wherein the truncation mutant comprises a deletion of 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, or 150, C-terminal amino acids of the full length diphtheria toxin.

15. The immunotoxin of claim 1 , wherein the diabody is not a fold-back diabody.

16. An immunotoxin, comprising a mutant diphtheria toxin moiety linked to a targeting moiety, wherein the targeting moiety is a single chain variable region diabody, and wherein the immunotoxin is AdmDT390scfbDb(J591) where the furin site amino acid sequence cleavage site RVRRSV (SEQ ID NO: 24) has been replaced with the MMP cleavage site GPLGMLSQ (SEQ ID NO: 25).

17. The immunotoxin of claim 16 , wherein the mutant diphtheria toxin moiety is a truncation mutant.

18. An immunotoxin fusion protein comprising a diphtheria toxin moiety and a targeting moiety, wherein the targeting moiety is a diabody, wherein the sequence from the amino terminus from left to right is toxin moiety,V L1 , L 2 , V H1 , L 3 , V L2 , L 4 , V H2 , wherein the toxin moiety comprises a truncation mutation, L 2 , L 3 and L 4 are (G 4 S) 3 linkers, and V L and V H are the variable light and heavy domains, wherein the V L1 and V H1 comprise the V L V H region of an anti-PSMA antibody, and wherein the V L1 and V H1 and the V L2 and V H2 share the same antigen specificity.

Assignees

• Neville Jr David M
• Woo Jung-Hee
• Frankel Arthur
• Us Health
• Scott And White Healthcare

Inventors

• Neville Jr David M
• Woo Jung-Hee
• Frankel Arthur

Classifications

• A61K47/6849

  the antibody targeting a receptor, a cell surface antigen or a cell surface determinant · CPC title

• A61P37/06

  Immunosuppressants, e.g. drugs for graft rejection · CPC title

• A61K47/6869

  the tumour determinant being from a cell of the reproductive system: ovaria, uterus, testes, prostate · CPC title

• A61K47/6855

  the tumour determinant being from breast cancer cell · CPC title

• A61K47/6829 Primary

  Bacterial toxins, e.g. diphteria toxins or Pseudomonas exotoxin A · CPC title