US 11,697,684 B2
Tri-specific binding molecules that specifically bind to multiple cancer antigens
Ezio Bonvini, Rockville, MD (US); Paul A. Moore, Rockville, MD (US); Jonathan C. Li, Rockville, MD (US); Leslie S. Johnson, Rockville, MD (US); and Kalpana Shah, Rockville, MD (US)
Assigned to MACROGENICS, INC., Rockville, MD (US)
Filed by MACROGENICS, INC., Rockville, MD (US)
Filed on Feb. 28, 2020, as Appl. No. 16/805,105.
Application 16/805,105 is a division of application No. 15/313,765, granted, now 10,633,440, previously published as PCT/US2015/033081, filed on May 29, 2015.
Claims priority of provisional application 62/107,824, filed on Jan. 26, 2015.
Claims priority of provisional application 62/008,229, filed on Jun. 5, 2014.
Claims priority of provisional application 62/004,571, filed on May 29, 2014.
Prior Publication US 2020/0199223 A1, Jun. 25, 2020
This patent is subject to a terminal disclaimer.
Int. Cl. C07K 16/28 (2006.01); C07K 16/30 (2006.01)
CPC C07K 16/28 (2013.01) [C07K 16/2803 (2013.01); C07K 16/283 (2013.01); C07K 16/2809 (2013.01); C07K 16/2815 (2013.01); C07K 16/2827 (2013.01); C07K 16/2878 (2013.01); C07K 16/30 (2013.01); C07K 2317/31 (2013.01); C07K 2317/524 (2013.01); C07K 2317/526 (2013.01); C07K 2317/55 (2013.01); C07K 2317/56 (2013.01); C07K 2317/565 (2013.01); C07K 2317/622 (2013.01); C07K 2317/626 (2013.01); C07K 2317/73 (2013.01); C07K 2317/92 (2013.01); C07K 2318/20 (2013.01); C07K 2319/00 (2013.01)] 13 Claims
 
1. A multi-chain polypeptide-containing Tri-Specific Binding Molecule that immunospecifically binds to three different epitopes, comprising:
(I) three different polypeptide chains covalently complexed together;
(II) an Antigen-Binding Domain I that immunospecifically binds to an Epitope I present on a first antigen, an Antigen-Binding Domain II that immunospecifically binds to an Epitope II present on a second antigen, and an Antigen-Binding Domain III that immunospecifically binds to an Epitope III present on a third antigen; and
(III) a Fc Domain; wherein:
(A) one of Epitope I, Epitope II or Epitope III is an epitope of an Effector Cell Antigen, a second of Epitope I, Epitope II or Epitope III is an epitope of a first Cancer Antigen, and a third of Epitope I, Epitope II or Epitope III is an epitope of a second Cancer Antigen;
(B) a first polypeptide chain comprises, in the N-terminus to C-terminus direction:
(VLI Domain)-(Linker 1)-(VHII Domain)-(Linker 2)-(Heterodimer-Promoting Domain)-(Linker 3)-(CH2-CH3 Domain); or
(Cysteine-Containing Domain)-(CH2-CH3 Domain)-(Linker 4)-(VLI Domain)-(Linker 1)-(VHII Domain)-(Linker 2)-(Heterodimer-Promoting Domain);
(C) a second polypeptide chain comprises, in the N-terminus to C-terminus direction:
(VLII Domain)-(Linker 1)-(VHI Domain)-(Linker 2)-(Heterodimer-Promoting Domain);
(D) a third polypeptide chain comprises, in the N-terminus to C-terminus direction:
(VLIII Domain)-(Flexible Linker)-(VHIII Domain)-(Cysteine-Containing Domain)-(CH2-CH3 Domain); or
(CL Domain)-(VLIII Domain)-(Flexible Linker)-(VHIII Domain)-(CH1 Domain)-(Cysteine-Containing Domain)-(CH2-CH3 Domain);
(E) the VLI Domain is a Light Chain Variable Domain of an immunoglobulin that binds to Epitope I, the VHI Domain is a Heavy Chain Variable Domain of an immunoglobulin that binds to Epitope I, the VLII Domain is a Light Chain Variable Domain of an immunoglobulin that binds to Epitope II, the VHII Domain is a Heavy Chain Variable Domain of an immunoglobulin that binds to Epitope II, the VLIII Domain is a Light Chain Variable Domain of an immunoglobulin that binds to Epitope III, and the VHIII Domain is a Heavy Chain Variable Domain of an immunoglobulin that binds to Epitope III;
(F) the VLI Domain and the VHI Domain associate to form the Antigen-Binding Domain I, the VLII Domain and the VHII Domain associate to form the Antigen-Binding Domain II, the VLIII Domain and the VHIII Domain associate to form the Antigen-Binding Domain III, and the CH2-CH3 Domain of the first polypeptide chain and the CH2-CH3 Domain of the third polypeptide chain associate to form the Fc Domain; and
(G) the Linker 1 comprises the sequence of SEQ ID NO: 33;
the Linker 2 comprises the sequence of SEQ ID NO: 34 or 47;
the Linker 3 comprises the sequence of SEQ ID NO: 46, 47, 48, 49, 50, 51, 152, or GCG or GGG;
the Linker 4 comprises the sequence of SEQ ID NO: 50 or 152;
the Heterodimer-Promoting Domain on the first polypeptide chain is an E-coil Domain and the Heterodimer-Promoting Domain on the second polypeptide chain is a K-coil Domain, or the Heterodimer-Promoting Domain on the first polypeptide chain is a K-coil Domain and the Heterodimer-Promoting Domain on the second polypeptide chain is an E-coil Domain, the E-coil Domain independently comprises the sequence of SEQ ID NO: 39 or 41, and the K-coil Domain independently comprises the sequence of SEQ ID NO: 40 or 42;
the Cysteine-Containing Domain independently comprises the sequence of SEQ ID NO: 34, 36, 38, 48, 210 or 211;
the CH1 Domain comprises the sequence of SEQ ID NO: 207 or 208; and
the CL Domain comprises the sequence of SEQ ID NO: 210 or 211.