Therapeutic compounds and methods
This disclosure describes engineered compounds that engage NK cells and methods of using the compounds. Generally, the compound includes an NK engaging domain, a targeting domain that selectively binds to a target cell, and an NK activating domain operably linking the NK engaging domain and the targeting domain.
1. A compound comprising:
an NK engaging domain comprising:
amino acid residues 1-240 of SEQ ID NO:1, or
amino acid residues 19-140 of SEQ ID NO:14;
an NK activating domain comprising:
the amino acid sequence of SEQ ID NO:15, or the amino acid sequence of SEQ ID NO:15 comprising an N72D amino acid substitution or an N72A amino acid substitution;
a first linker linking the NK engaging domain to the NK activating domain;
a targeting domain that selectively binds to a target antigen; and
a second linker linking the NK activating domain to the targeting domain.
2. The compound of claim 1 , wherein the NK engaging domain activates an NK cell.
3. The compound of claim 1 , wherein the NK engaging domain blocks inhibition of an NK cell.
4. The compound of claim 1 wherein the NK engaging domain moiety comprises an antibody or a binding fragment thereof.
5. The compound of claim 4 wherein the antibody fragment comprises an scFv, a F(ab)2, or a Fab.
6. The compound of claim 1 wherein the targeting domain comprises a moiety that selectively binds to a tumor cell.
7. The compound of claim 6 wherein the targeting domain moiety comprises an antibody or a binding fragment thereof.
8. The compound of claim 7 wherein the antibody fragment comprises an scFv.
9. A composition comprising:
the compound of claim 1 ; and
a pharmaceutically acceptable carrier.
10. A compound comprising the amino acid sequence set forth in SEQ ID NO:1 or SEQ ID NO:14.
11. The compound of claim 10 , wherein the compound comprises two targeting domains.
12. The compound of claim 1 , wherein the NK engager domain and the IL-15 sequence are linked by a linker as set forth in SEQ ID NO:3.
13. The compound of claim 1 , wherein the IL-15 sequence and the targeting domain are linked by a linker as set forth in SEQ ID NO:4.
14. The compound of claim 5 , wherein the antibody, the binding fragment thereof, or the nanobody is human, humanized, or camelid.
15. The compound of claim 1 , wherein the first linker comprises:
the amino acids of SEQ ID NO:3;
the amino acids of SEQ ID NO:4;
amino acids 119-133 of SEQ ID NO:5;
amino acids 494-518 of SEQ ID NO:8;
amino acids 241-255 of SEQ ID NO:12;
amino acids 488-506 of SEQ ID:10;
amino acids 277-294 of SEQ ID NO:14; or
amino acids 142-161 of SEQ ID NO:14.
16. The compound of claim 1 , wherein the second linker comprises:
the amino acids of SEQ ID NO:3;
the amino acids of SEQ ID NO:4;
amino acids 119-133 of SEQ ID NO:5;
amino acids 494-518 of SEQ ID NO:8;
amino acids 241-255 of SEQ ID NO:12;
amino acids 488-506 of SEQ ID:10;
amino acids 277-294 of SEQ ID NO:14; or amino acids 142-161 of SEQ ID NO:14.
17. The compound of claim 1 , wherein:
the first linker comprises the amino acids of SEQ ID NO:3; and
the second linker comprises the amino acids of SEQ ID NO:4.
18. The compound of claim 1 , wherein:
the first linker comprises amino acids 142-161 of SEQ ID NO:14; and
the second linker comprises amino acids 277-294 of SEQ ID NO:14.
19. The compound of claim 1 , wherein the targeting domain binds to a target antigen or portion thereof selected from CD133, CD20, H ER2, CEA, EpCAM, VEGF-A, EGFR, CD33, integrin αVβ3, CD51, CD152, CD125, CTAA16.88, MUC1, CD19, CD22, CD38, CD30, CD52, uPAR, LIV-1, CD70, IL-3, IL-4R, mesothelin, ROR1, CSPG4, SS1, IGFR1, HSPG2, IGF-1, epithelial-mesenchymal transition (EMT), TRAIL, CD45, CD74, CD23, HIV, or a cancer antigen.