CD38-binding proteins comprising de-immunized Shiga toxin A subunit effectors

Patent metadata

Field	Value
Publication number	US-11713347-B2
Application number	US-202016751144-A
Country	US
Kind code	B2
Filing date	Jan 23, 2020
Priority date	Jan 23, 2019
Publication date	Aug 1, 2023
Grant date	Aug 1, 2023

Abstract

The instant invention provides binding proteins (“CD38-binding proteins”) which each comprise (1) a CD38-binding region for cell-targeting and (2) a Shiga toxin A Subunit effector polypeptide (“Shiga toxin effector polypeptide”). The Shiga toxin effector polypeptide components of the CD38-binding proteins may comprise a combination of mutations relative to a wild-type Shiga toxin sequence providing (1) de-immunization and/or (2) a reduction in protease sensitivity; wherein each Shiga toxin effector polypeptide retains one or more Shiga toxin function, such as, e.g., stimulating cellular internalization, directing intracellular routing, catalytic activity, and/or potent cytotoxicity. The CD38-binding proteins may have one or multiple uses, e.g., the selective killing of a specific CD38-expressing cell-type; and more generally, for the diagnosis and treatment of cancers and disorders involving CD38-expressing cells, e.g., in CD38-positive hematopoietic cancers such as multiple myeloma.

First claim

What is claimed is:

1. A CD38-binding fusion protein comprising:
1) a Shiga toxin A subunit effector polypeptide; and
2) a CD38-binding domain, comprising:
a) a heavy chain variable domain (VH) comprising:
i) a vHCDR1 comprising the sequence of SEQ ID NO: 34;
ii) a vHCDR2 comprising the sequence of SEQ ID NO: 35; and
iii) a vHCDR3 comprising the sequence of SEQ ID NO: 36; and

b) a light chain variable domain (VL) comprising:
i) a vLCDR1 comprising the sequence of SEQ ID NO: 31;
ii) a vLCDR2 comprising the sequence of SEQ ID NO: 32; and
iii) a vLCDR3 comprising the sequence of SEQ ID NO: 33.

2. The CD38-binding fusion protein of claim 1 , wherein the CD38-binding fusion protein comprises the sequence of SEQ ID NO: 79.

3. The CD38-binding fusion protein of claim 1 , wherein the VL comprises the sequence of SEQ ID NO: 43, and the VH comprises the sequence of SEQ ID NO: 44.

4. The CD38-binding fusion protein of claim 1 , wherein the CD38-binding fusion protein comprises a first linker between the Shiga toxin A subunit effector polypeptide and the CD38-binding domain.

5. The CD38-binding fusion protein of claim 4 , wherein the first linker is a proteinaceous linker.

6. The CD38-binding fusion protein of claim 5 , wherein the first linker comprises the sequence of SEQ ID NO: 70.

7. The CD38-binding fusion protein of claim 4 , wherein the CD38-binding fusion protein comprises a second linker between the VH and the VL.

8. The CD38-binding fusion protein of claim 7 , wherein the CD38-binding fusion protein comprises, from its N- to C-terminus, the Shiga toxin A subunit effector polypeptide-first linker-VH-second linker-VL.

9. The CD38-binding fusion protein of claim 8 , wherein the Shiga toxin A subunit effector polypeptide comprises the sequence of SEQ ID NO: 46.

10. The CD38-binding fusion protein of claim 1 , wherein the Shiga toxin A subunit effector polypeptide comprises the sequence of SEQ ID NO: 46, the VL comprises the sequence of SEQ ID NO: 43, and the VH comprises the sequence of SEQ ID NO: 44.

11. The CD38-binding fusion protein of claim 1 , wherein the CD38-binding fusion protein is a homodimer.

12. The CD38-binding fusion protein of claim 1 , wherein the CD38-binding fusion protein comprises two identical polypeptides, each polypeptide comprising the sequence of SEQ ID NO: 79.

13. A nucleic acid encoding the CD38-binding protein fusion of claim 2 .

14. An expression vector comprising the nucleic acid of claim 13 .

15. A host cell comprising the expression vector of claim 14 .

16. A composition comprising (i) the CD38-binding fusion protein of claim 2 and (ii) a pharmaceutically acceptable carrier or excipient.

17. The composition of claim 16 , wherein at least about 90% of the CD38-binding fusion protein is in the form of a homodimer.

18. The composition of claim 17 , wherein at least about 95% of the CD38-binding fusion protein is in the form of a homodimer.

Assignees

• Milennium Pharmaceuticals Inc
• Molecular Templates Inc
• Millennium Pharm Inc

Inventors

• Chattopadhyay Nibedita
• Poma Eric
• Willert Erin

Classifications

• C07K14/70521 Primary

  CD28, CD152 · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• C12N15/63

  Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression · CPC title

• A61K38/00

  Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) · CPC title

• C07K2317/565

  Complementarity determining region [CDR] · CPC title