Binding fusion proteins, binding fusion protein-drug conjugates, XTEN-drug conjugates and methods of making and using same
The present invention relates to binding fusion protein compositions comprising targeting moieties linked to extended recombinant polypeptide (XTEN), binding fusion protein-drug conjugate compositions, and XTEN-drug conjugate compositions, isolated nucleic acids encoding the compositions and vectors and host cells containing the same, and methods of using such compositions in treatment of diseases, disorders, and conditions.
1. A binding fusion protein comprising:
a first targeting moiety comprising a single-chain variable fragment (scFv) with specific binding affinity to CD3; and
a first extended recombinant polypeptide (XTEN), wherein the first XTEN comprises a motif selected from the group consisting of SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, and SEQ ID NO: 9.
2. The binding fusion protein of claim 1 , wherein the first XTEN comprises an amino acid sequence which has at least 90% sequence identity to a sequence selected from the group consisting of AE144 (SEQ ID NO: 41), AE288 (SEQ ID NO: 47), AE576 (SEQ ID NO: 54) and AE864 (SEQ ID NO: 59).
3. The binding fusion protein of claim 2 , wherein the first XTEN comprises an amino acid sequence selected from the group consisting of AE144 (SEQ ID NO: 41), AE288 (SEQ ID NO: 47), AE576 (SEQ ID NO: 54) and AE864 (SEQ ID NO: 59).
4. The binding fusion protein of claim 1 , wherein the first targeting moiety has specific binding affinity for an epsilon subunit of CD3 (CDR).
5. The binding fusion protein of claim 1 , wherein the first XTEN is positioned N-terminal of the first targeting moiety.
6. The binding fusion protein of claim 1 , wherein the first XTEN is positioned C-terminal of the first targeting moiety.
7. The binding fusion protein of claim 1 , further comprising a second targeting moiety that differs from the first targeting moiety.
8. The binding fusion protein of claim 1 , wherein the binding fusion protein is configured according to formula I:
(XTEN) x -TM-(XTEN) y I
wherein x is either 0 or 1; y is either 0 or 1, wherein x+y≥1, and wherein TM is the first targeting moiety.
9. The binding fusion protein of claim 8 , wherein x is 1 and y is 1.
10. The binding fusion protein of claim 7 , further comprising a second XTEN, wherein the second XTEN comprises a motif selected from the group consisting of SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, and SEQ ID NO: 9.
11. The binding fusion protein of claim 10 , wherein the second XTEN comprises an amino acid sequence which has at least 90% sequence identity to a sequence selected from the group consisting of AE144 (SEQ ID NO: 41), AE288 (SEQ ID NO: 47), AE576 (SEQ ID NO: 54) and AE864 (SEQ ID NO: 59).
12. The binding fusion protein of claim 11 , wherein the second XTEN comprises an amino acid sequence selected from the group consisting of AE144 (SEQ ID NO: 41), AE288 (SEQ ID NO: 47), AE576 (SEQ ID NO: 54) and AE864 (SEQ ID NO: 59).
13. The binding fusion protein of claim 10 , wherein the first XTEN is positioned at the N-terminus of the binding fusion protein and the second XTEN is positioned at the C-terminus of binding fusion protein.
14. A pharmaceutical composition comprising the binding fusion protein of claim 1 , and at least one pharmaceutically acceptable carrier.
15. The pharmaceutical composition of claim 14 , wherein the first XTEN comprises an amino acid sequence which has at least 90% sequence identity to a sequence selected from the group consisting of AE144 (SEQ ID NO: 41), AE288 (SEQ ID NO: 47), AE576 (SEQ ID NO: 54) and AE864 (SEQ ID NO: 59).
16. A kit comprising:
packaging material;
at least a first container comprising the pharmaceutical composition of claim 14 ;
a label identifying the pharmaceutical composition and providing storage and handling conditions; and
a sheet of instructions for the reconstitution and/or administration of the pharmaceutical composition to a subject.
17. A method of treating a human disease, the method comprising administering a therapeutically effective dose of the pharmaceutical composition of claim 14 to direct cytotoxic activity of T-cells towards tumor cells.
18. The method of claim 17 , wherein the human disease is selected from the group consisting of breast carcinoma, lung carcinoma, gastric carcinoma, esophageal carcinoma, colorectal carcinoma, liver carcinoma, ovarian carcinoma, thecoma, arrhenoblastoma, cervical carcinoma, endometrial carcinoma, fibrosarcoma, choriocarcinoma, head and neck cancer, nasopharyngeal carcinoma, laryngeal carcinoma, hepatoblastoma, Kaposi's sarcoma, melanoma, skin carcinoma, hemangioma, cavernous hemangioma, hemangioblastoma, pancreas carcinoma, retinoblastoma, astrocytoma, glioblastoma, Schwannoma, oligodendroglioma, medulloblastoma, neuroblastoma, rhabdomyosarcoma, osteogenic sarcoma, leiomyosarcomas, urinary tract carcinoma, thyroid carcinoma, Wilm's tumor, renal cell carcinoma, and prostate carcinoma.
19. The isolated binding fusion protein of claim 7 , wherein the second targeting moiety comprises a second single-chain variable fragment (scFv).