Glypican-3-binding fibronectin based scaffold molecules
Provided herein are polypeptides which include tenth fibronectin type III domains ( 10 Fn3) that bind to glypican-3. Also provided are fusion molecules comprising a 10 Fn3 domain that bind to glypican-3 for use in diagnostic and therapeutic applications. Glypican-3 10 Fn3 drug conjugates are also provided.
1. A nucleic acid encoding a polypeptide comprising a tenth fibronectin type III ( 10 Fn3) domain comprising BC, DE and FG loops, wherein the polypeptide binds specifically to human glypican-3 (GPC3), and wherein
(a) the BC, DE and FG loops comprise SEQ ID NOs: 6, 7 and 8, respectively;
(b) the BC, DE and FG loops comprise SEQ ID NOs: 19, 20 and 21, respectively;
(c) the BC, DE and FG loops comprise SEQ ID NOs: 32, 33 and 34, respectively;
(d) the BC, DE and FG loops comprise SEQ ID NOs: 45, 46 and 47, respectively;
(e) the BC, DE and FG loops comprise SEQ ID NOs: 58, 59 and 60, respectively;
(f) the BC, DE and FG loops comprise SEQ ID NOs: 71, 72 and 73, respectively;
(g) the BC, DE and FG loops comprise SEQ ID NOs: 84, 85 and 86, respectively;
(h) the BC, DE and FG loops comprise SEQ ID NOs: 99, 100 and 101, respectively;
(i) the BC, DE and FG loops comprise SEQ ID NOs: 99, 100 and 129, respectively;
(j) the BC, DE and FG loops comprise SEQ ID NOs: 99, 100 and 156, respectively;
(k) the BC, DE and FG loops comprise SEQ ID NOs: 99, 100 and 183, respectively;
(l) the BC, DE and FG loops comprise SEQ ID NOs: 99, 100 and 210, respectively;
(m) the BC, DE and FG loops comprise SEQ ID NOs: 99, 100 and 237, respectively;
(n) the BC, DE and FG loops comprise SEQ ID NOs: 99, 100 and 264, respectively;
(o) the BC, DE and FG loops comprise SEQ ID NOs: 99, 100 and 291, respectively; or
(p) the BC, DE and FG loops comprise SEQ ID NOs: 99, 100 and 318, respectively.
2. The nucleic acid of claim 1 , wherein the nucleic acid encodes a polypeptide comprising an amino acid sequence at least 90% to the amino acid sequence of any one of SEQ ID NOs: 5, 9-18, 22-31, 35-44, 48-57, 61-70, 74-83, 87-98, 102-128, 130-155, 157-182, 184-209, 211-236, 238-263, 265-290, 292-317 or 319-343.
3. The nucleic acid of claim 1 , wherein the nucleic acid encodes a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 5, 9-18, 22-31, 35-44, 48-57, 61-70, 74-83, 87-98, 102-128, 130-155, 157-182, 184-209, 211-236, 238-263, 265-290, 292-317 and 319-343.
4. The nucleic acid of claim 1 , wherein the nucleic acid encodes a polypeptide further comprising a heterologous protein.
5. The nucleic acid of claim 4 , wherein the heterologous protein comprises a polypeptide selected from the group consisting of a 10 Fn3 domain, an Fc, Fc fragment, transferrin, serum albumin, a serum albumin binding protein, and a serum immunoglobulin binding protein.
6. The nucleic acid of claim 1 , wherein the C-terminus of the 10 Fn3 domain comprises a moiety consisting of the amino acid sequence P m X n , wherein P is proline, each X is independently any amino acid, m is an integer that is at least 1 and n is 0 or an integer that is at least 1.
7. The nucleic acid of claim 6 , wherein the C-terminal moiety comprises cysteine.
8. A pharmaceutical composition comprising the nucleic acid of claim 1 , and a pharmaceutically acceptable carrier.
9. The nucleic acid of claim 1 , wherein the nucleic acid encodes a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 98 and 102-127.
10. The nucleic acid of claim 1 , wherein the nucleic acid encodes a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 263 and 265-289.
11. A vector comprising the nucleic acid of claim 1 .
12. A host cell line expressing a recombinant polypeptide encoded by the nucleic acid of claim 1 .