US 12,227,573 B2
Trispecific binding proteins, methods, and uses thereof
Christian Beil, Frankfurt am Main (DE); Jochen Beninga, Frankfurt Am Main (DE); Joerg Birkenfeld, Frankfurt Am Main (DE); Gary J. Nabel, Cambridge, MA (US); Huawei Qiu, Westborough, MA (US); Ercole Rao, Morfelden-Waldorf (DE); Joerg Regula, Munich (DE); Edward Seung, Cambridge, MA (US); Ronnie Wei, Needman, MA (US); Lan Wu, Cambridge, MA (US); Zhen Xing, Cambridge, MA (US); Ling Xu, Cambridge, MA (US); Zhi-Yong Yang, Cambridge, MA (US); Béatrice Cameron, Paris (FR); Tarik Dabdoubi, Paris (FR); Cendrine Lemoine, Paris (FR); and Catherine Prades, Paris (FR)
Assigned to Sanofi, Paris (FR)
Filed by Sanofi, Paris (FR)
Filed on Mar. 13, 2023, as Appl. No. 18/183,107.
Application 18/183,107 is a division of application No. 16/843,792, filed on Apr. 8, 2020, granted, now 11,613,576.
Claims priority of provisional application 62/831,572, filed on Apr. 9, 2019.
Claims priority of provisional application 62/831,415, filed on Apr. 9, 2019.
Claims priority of application No. 19306261 (EP), filed on Oct. 2, 2019; and application No. 19306312 (EP), filed on Oct. 8, 2019.
Prior Publication US 2023/0220079 A1, Jul. 13, 2023
Int. Cl. C07K 16/28 (2006.01); A61K 39/395 (2006.01); A61K 45/06 (2006.01); A61P 35/00 (2006.01); C07K 16/32 (2006.01); C07K 16/46 (2006.01); C12N 15/63 (2006.01)
CPC C07K 16/2809 (2013.01) [A61K 39/3955 (2013.01); A61K 39/39558 (2013.01); A61K 45/06 (2013.01); A61P 35/00 (2018.01); C07K 16/2818 (2013.01); C07K 16/2896 (2013.01); C07K 16/32 (2013.01); C07K 16/468 (2013.01); C12N 15/63 (2013.01); C07K 2317/31 (2013.01); C07K 2317/52 (2013.01); C07K 2317/522 (2013.01); C07K 2317/524 (2013.01); C07K 2317/526 (2013.01); C07K 2317/53 (2013.01); C07K 2317/565 (2013.01)] 45 Claims
 
1. A method of treating cancer in a patient comprising administering to the patient a therapeutically effective amount of a binding protein, wherein the binding protein comprises four polypeptide chains that form three antigen binding sites, wherein a first polypeptide chain comprises a structure represented by the formula:
VL2-L1-VL1-L2-CL  [I]
and a second polypeptide chain comprises a structure represented by the formula:
VH1-L3-VH2-L4-CH1-hinge-CH2-CH3  [II]
and a third polypeptide chain comprises a structure represented by the formula:
VH3-CH1-hinge-CH2-CH3  [III]
and a fourth polypeptide chain comprises a structure represented by the formula:
VL3-CL  [IV]
wherein:
VL1 is a first immunoglobulin light chain variable domain;
VL2 is a second immunoglobulin light chain variable domain;
VL3 is a third immunoglobulin light chain variable domain;
VH1 is a first immunoglobulin heavy chain variable domain;
VH2 is a second immunoglobulin heavy chain variable domain;
VH3 is a third immunoglobulin heavy chain variable domain;
CL is an immunoglobulin light chain constant domain;
CH1 is an immunoglobulin CH1 heavy chain constant domain;
CH2 is an immunoglobulin CH2 heavy chain constant domain;
CH3 is an immunoglobulin CH3 heavy chain constant domain;
hinge is an immunoglobulin hinge region connecting the CH1 and CH2 domains; and L1,
L2, L3 and L4 are amino acid linkers;
wherein the polypeptide of formula I and the polypeptide of formula II form a cross-over light chain-heavy chain pair; and
wherein VH1 and VL1 form a first antigen binding site;
wherein VH2 and VL2 form a second antigen binding site that binds a CD3 polypeptide, wherein the VH2 domain comprises a CDR-H1 sequence comprising the amino acid sequence of GFTFTKAW (SEQ ID NO:55), a CDR-H2 sequence comprising the amino acid sequence of IKDKSNSYAT (SEQ ID NO:56), and a CDR-H3 sequence comprising the amino acid sequence of RGVYYALSPFDY (SEQ ID NO:57), and the VL2 domain comprises a CDR-L1 sequence comprising the amino acid sequence of QSLVHX1NX2X3TY, wherein X1 is E or Q, X2 is A or L, and X3 is Q, R, or F (SEQ ID NO:180), a CDR-L2 sequence comprising the amino acid sequence of KVS, and a CDR-L3 sequence comprising the amino acid sequence of GQGTQYPFT (SEQ ID NO:65);
wherein VH3 and VL3 form a third antigen binding site; and
wherein the third antigen binding site binds a tumor target protein.