Method of treating cancer by administering EGFR and EGFR/IGFIR binding molecules
The present invention relates to bispecific molecules comprising an EGFR binding domain and a distinct IGFIR binding domain for use in diagnostic, research and therapeutic applications. The invention further relates to cells comprising such proteins, polynucleotide encoding such proteins or fragments thereof, and vectors comprising the polynucleotides encoding the innovative proteins. Exemplary bispecific molecules include antibody-like protein dimers based on the tenth fibronectin type III domain.
1. A method for treating a cancer associated with epidermal growth factor receptor (EGFR) signaling in a subject comprising administering to a subject in need thereof a therapeutically effective amount of an antibody-like protein comprising a tenth fibronectin type III domain ( 10 Fn3) that binds EGFR with a K D of less than 500 nM, wherein the EGFR binding 10 Fn3 comprises a BC loop having the amino acid sequence X g WAPVDRYQX h (SEQ ID NO: 137), a DE loop having the amino acid sequence X i RDVYX j (SEQ ID NO: 138), and an FG loop having the amino acid sequence X k DYKPHADGPHTYHESX l (SEQ ID NO: 139); and
wherein X is any amino acid and a, b, c, d, e, f, g, h, i, j, k, and l are integers independently selected from 0 to 5.
2. The method of claim 1 , wherein the EGFR binding 10 Fn3 comprises a BC loop having the amino acid sequence SWWAPVDRYQ (SEQ ID NO: 115), a DE loop having the amino acid sequence PRDVYT (SEQ ID NO: 116), and an FG loop having the amino acid sequence TDYKPHADGPHTYHESP (SEQ ID NO: 117).
3. The method of claim 1 , wherein the antibody-like protein further comprises a second tenth fibronectin type III domain ( 10 Fn3).
4. The method of claim 3 , wherein the second 10 Fn3 is covalently linked to the EGFR binding 10 Fn3 via a polypeptide linker or a polyethylene glycol moiety.
5. The method of claim 3 , wherein the second 10 Fn3 binds to IGF-IR.
6. The method of claim 5 , wherein the IGF-IR binding 10 Fn3 comprises a BC loop having the amino acid sequence X a SARLKVAX b (SEQ ID NO: 46), a DE loop having the amino acid sequence X c KNVYX d (SEQ ID NO: 48), and an FG loop having the amino acid sequence X e RFRDYQX f (SEQ ID NO: 50).
7. The method of claim 6 , wherein the IGF-IR binding 10 Fn3 comprises a BC loop having the amino acid sequence SWSARLKVAR (SEQ ID NO: 45), a DE loop having the amino acid sequence PKNVYT (SEQ ID NO: 47), and an FG loop having the amino acid sequence TRFRDYQP (SEQ ID NO: 49).
8. The method of claim 1 , wherein the antibody-like protein further comprises one or more pharmacokinetic (PK) moieties.
9. The method of claim 1 , wherein the cancer is selected from the group consisting of: brain cancer, urogenital tract cancer, lymphatic cancer, stomach cancer, laryngeal cancer, monocytic leukemia, lung adenocarcinoma, small-cell lung carcinoma, pancreatic cancer, glioblastoma, squamous cell carcinoma, bladder cancer, liver cancer, kidney cancer, colorectal cancer, breast cancer, head cancer, neck cancer, oesophageal cancer, gynecological cancer, thyroid cancer, lymphoma, chronic leukemia, and acute leukemia.
10. The method of claim 1 , further comprising administering one or more additional therapeutic agents.
11. The method of claim 10 , wherein the one or more additional therapeutic agents is a cancer therapeutic agent.
12. A method of treating a cancer associated with epidermal growth factor receptor (EGFR) and/or insulin-like growth factor 1 receptor (IGF-1R) signaling in a subject comprising administering to a subject in need thereof a therapeutically effective amount of an antibody-like protein comprising:
(a) a tenth fibronectin type III domain ( 10 Fn3) that binds EGFR with a K D of less than 500 nM, wherein the EGFR binding 10 Fn3 comprises a BC loop having the amino acid sequence X g WAPVDRYQX h (SEQ ID NO: 137), a DE loop having the amino acid sequence X i RDVYX j (SEQ ID NO: 138), and an FG loop having the amino acid sequence X k DYKPHADGPHTYHESX l (SEQ ID NO: 139), and
(b) a 10 Fn3 that binds IGF-1R comprising a BC loop having the amino acid sequence X a SARLKVAX b (SEQ ID NO: 46), a DE loop having the amino acid sequence X c KNVYX d (SEQ ID NO: 48), and an FG loop having the amino acid sequence X e RFRDYQX f (SEQ ID NO: 50),
wherein X is any amino acid and a, b, c, d, e, f, g, h, i, j, k, and l are integers independently selected from 0 to 5.
13. The method of claim 12 , wherein the IGF-IR binding 10 Fn3 comprises a BC loop having the amino acid sequence SWSARLKVAR (SEQ ID NO: 45), a DE loop having the amino acid sequence PKNVYT (SEQ ID NO: 47), and an FG loop having the amino acid sequence TRFRDYQP (SEQ ID NO: 49).
14. The method of claim 13 , wherein the EGFR binding 10 Fn3 has an amino acid sequence at least 90% identical to SEQ ID NO: 112, and the IGF-IR binding 10 Fn3 has an amino acid sequence at least 90% identical to SEQ ID NO: 3.
15. The method of claim 14 , wherein the EGFR binding 10 Fn3 comprises the amino acid sequence set forth in SEQ ID NO: 112 and the IGF-IR binding 10 Fn3 comprises the amino acid sequence set forth in SEQ ID NO: 3.
16. The method of claim 13 , wherein the antibody-like protein comprises an amino acid sequence at least 90% identical to any one of SEQ ID NOs: 126-132.
17. The method of claim 16 , wherein the antibody-like protein comprises an amino acid sequence at least 90% identical to SEQ ID NO: 130.
18. The method of claim 17 , wherein the antibody-like protein comprises the amino acid sequence of SEQ ID NO: 130.
19. The method of claim 12 , wherein the antibody-like protein further comprises one or more pharmacokinetic (PK) moieties.
20. The method of claim 12 , further comprising administering one or more additional therapeutic agents.