US 12,215,157 B2
Multi-specific binding proteins that bind CD33, NKG2D, and CD16, and methods of use
Gregory P. Chang, Medford, MA (US); Ann F. Cheung, Lincoln, MA (US); Asya Grinberg, Lexington, MA (US); Dhruv Kam Sethi, Belmont, MA (US); William Haney, Wayland, MA (US); Bianka Prinz, Lebanon, NH (US); Bradley M. Lunde, Lebanon, NH (US); Ronnie Wei, Weston, MA (US); Daniel Fallon, Winchester, MA (US); and Steven O'Neil, Wayland, MA (US)
Assigned to Dragonfly Therapeutics, Inc., Waltham, MA (US)
Appl. No. 16/971,104
Filed by Dragonfly Therapeutics, Inc., Waltham, MA (US)
PCT Filed Feb. 20, 2019, PCT No. PCT/US2019/018751
§ 371(c)(1), (2) Date Aug. 19, 2020,
PCT Pub. No. WO2019/164930, PCT Pub. Date Aug. 29, 2019.
Claims priority of provisional application 62/632,756, filed on Feb. 20, 2018.
Claims priority of provisional application 62/677,137, filed on May 28, 2018.
Prior Publication US 2021/0101976 A1, Apr. 8, 2021
Int. Cl. C07K 16/28 (2006.01)
CPC C07K 16/283 (2013.01) [C07K 2317/31 (2013.01); C07K 2317/522 (2013.01); C07K 2317/55 (2013.01); C07K 2317/565 (2013.01); C07K 2317/622 (2013.01); C07K 2317/92 (2013.01)] 40 Claims
 
1. A protein comprising:
(a) a first antigen-binding site that binds human NKG2D;
(b) a second antigen-binding site that binds human CD33; and
(c) an antibody Fc domain that binds human CD16 or a portion thereof that binds human CD16,
wherein the first antigen-binding site that binds human NKG2D comprises a Fab fragment, and
wherein the Fab fragment comprises a light chain complementarity-determining region 1 (CDR1) sequence comprising the amino acid sequence of SEQ ID NO:114, a light chain complementarity-determining region 2 (CDR2) sequence comprising the amino acid sequence of SEQ ID NO:115, a light chain complementarity-determining region 3 (CDR3) sequence comprising the amino acid sequence of SEQ ID NO: 116, a heavy chain complementarity-determining region 1 (CDR1) sequence comprising the amino acid sequence of SEQ ID NO:111, a heavy chain complementarity-determining region 2 (CDR2) sequence comprising the amino acid sequence of SEQ ID NO:112, and a heavy chain complementarity-determining region 3 (CDR3) sequence comprising the amino acid sequence of SEQ ID NO:113;
and wherein the second antigen-binding site that binds human CD33 comprises a single-chain variable fragment (scFv), and wherein the scFv comprises a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:48, a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:49, a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:50, a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:45, a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:46, and a heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:47.