US 12,378,318 B2
Proteins binding NKG2D, CD16 and a tumor-associated antigen
Gregory P. Chang, Medford, MA (US); Ann F. Cheung, Lincoln, MA (US); Jinyan Du, Waltham, MA (US); Daniel Fallon, Winchester, MA (US); Asya Grinberg, Lexington, MA (US); William Haney, Wayland, MA (US); Bradley M. Lunde, Lebanon, NH (US); Steven O'Neil, Wayland, MA (US); Bianka Prinz, Lebanon, NH (US); and Ronnie Wei, Weston, MA (US)
Assigned to DRAGONFLY THERAPEUTICS, INC., Waltham, MA (US)
Appl. No. 17/266,966
Filed by Dragonfly Therapeutics, Inc., Waltham, MA (US)
PCT Filed Aug. 8, 2019, PCT No. PCT/US2019/045677
§ 371(c)(1), (2) Date Feb. 8, 2021,
PCT Pub. No. WO2020/033664, PCT Pub. Date Feb. 13, 2020.
Claims priority of provisional application 62/716,110, filed on Aug. 8, 2018.
Prior Publication US 2022/0153848 A1, May 19, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C07K 16/28 (2006.01); A61P 35/00 (2006.01)
CPC C07K 16/2851 (2013.01) [A61P 35/00 (2018.01); C07K 16/283 (2013.01); C07K 16/2863 (2013.01); C07K 2317/31 (2013.01); C07K 2317/33 (2013.01); C07K 2317/52 (2013.01); C07K 2317/565 (2013.01); C07K 2317/76 (2013.01); C07K 2317/94 (2013.01)] 11 Claims
 
1. A trispecific binding protein comprising an Fc region that binds CD16 and two antigen binding sites, wherein the two antigen binding sites consist of a first antigen binding site that binds NKG2D and a second antigen binding site that binds cMET, wherein the protein comprises three polypeptides, and wherein
a) the first polypeptide comprises, from N-terminus to C-terminus,
i) a heavy chain variable domain of a Fab fragment that binds NKG2D,
ii) a CH1 domain,
iii) a first CH2 domain and
iv) a first CH3 domain;
b) the second polypeptide comprises, from N-terminus to C-terminus,
i) a light chain variable domain of the Fab fragment that binds NKG2D and
ii) a CL domain;
c) the third polypeptide comprises, from N-terminus to C-terminus,
i) an scFv that binds cMET,
ii) a second CH2 domain and
iii) a second CH3 domain,
(1) wherein the heavy chain variable domain of the Fab fragment that binds NKG2D comprises CDR1, CDR2, and CDR3 of amino acid sequence SEQ ID NO: 387, SEQ ID NO: 88, and SEQ ID NO: 390, respectively; and the light chain variable domain of the Fab fragment that binds NKG2D comprises CDR1, CDR2, and CDR3 of amino acid sequence SEQ ID NO: 90, SEQ ID NO: 91, and SEQ ID NO: 92, respectively; and
(2) wherein a heavy chain variable domain of the scFv that binds cMET comprises CDR1, CDR2, and CDR3 of amino acid sequence SEQ ID NO: 414, SEQ ID NO: 415, and SEQ ID NO: 416, respectively; and a light chain variable domain of the scFv that binds cMET comprises CDR1, CDR2, and CDR3 of amino acid sequence SEQ ID NO: 418, SEQ ID NO: 419, and SEQ ID NO: 420, respectively; and
(3) wherein the Fc region comprises the first CH2 domain, first CH3 domain, second CH2 domain and second CH3 domain.