Trispecific and/or trivalent binding proteins

Patent metadata

Field	Value
Publication number	US-2021061925-A1
Application number	US-202017099439-A
Country	US
Kind code	A1
Filing date	Nov 16, 2020
Priority date	Apr 13, 2016
Publication date	Mar 4, 2021
Grant date	—

Abstract

The disclosure provides trispecific and/or trivalent binding proteins comprising four polypeptide chains that form three antigen binding sites that specifically bind one or more target proteins, wherein a first pair of polypeptides forming the binding protein possess dual variable domains having a cross-over orientation and wherein a second pair of polypeptides forming the binding protein possess a single variable domain. The disclosure also provides methods for making trispecific and/or trivalent binding proteins and uses of such binding proteins.

First claim

1 - 90 . (canceled)

91 . A trivalent binding protein comprising four polypeptide chains that form three antigen binding sites that specifically bind one or more target proteins, wherein a first polypeptide chain of the binding protein comprises a structure represented by the formula:
V L2 -L 1 -V L1 -L 2 -C L   [I]

and a second polypeptide chain of the binding protein comprises a structure represented by the formula:
V H1 -L 3 -V H2 -L 4 -C H1 -hinge-C H2 -C H3   [II]

and a third polypeptide chain of the binding protein comprises a structure represented by the formula:
V H3 -C H1 -hinge-C H2 -C H3   [III]

and a fourth polypeptide chain of the binding protein comprises a structure represented by the formula:
V L3 -C L   [IV]

wherein:
V L1 is a first immunoglobulin light chain variable domain;
V L2 is a second immunoglobulin light chain variable domain;
V L3 is a third immunoglobulin light chain variable domain;
V H1 is a first immunoglobulin heavy chain variable domain;
V H2 is a second immunoglobulin heavy chain variable domain;
V H3 is a third immunoglobulin heavy chain variable domain;
C L is an immunoglobulin light chain constant domain;
C H1 is an immunoglobulin C H1 heavy chain constant domain;
C H2 is an immunoglobulin C H2 heavy chain constant domain;
C H3 is an immunoglobulin C H3 heavy chain constant domain;
hinge is an immunoglobulin hinge region connecting the C H1 and C H2 domains; and
L 1 , L 2 , L 3 and L 4 are each independently amino acid linkers or zero amino acids in length;
wherein the polypeptide of formula I and the polypeptide of formula II form a cross-over light chain-heavy chain pair of the binding protein; wherein V H1 and V L1 form a binding pair and a first antigen binding site, and V H2 and V L2 form a binding pair and a second antigen binding site; and wherein V H3 and V L3 form a binding pair and a third antigen binding site.

92 . The binding protein of claim 91 , wherein the binding protein is trivalent and capable of specifically binding one antigen target.

93 . The binding protein of claim 91 , wherein the binding protein is trivalent and capable of specifically binding two antigen targets.

94 . The binding protein of claim 91 , wherein the binding protein is trivalent and capable of specifically binding three antigen targets.

95 . The binding protein of claim 91 , wherein L 1 , L 2 , L 3 and L 4 each independently are the amino acid S or comprise a sequence selected from the group consisting of GGGGSGGGGS (SEQ ID NO:104), GGGGSGGGGSGGGGS (SEQ ID NO:105), RT, TKGPS (SEQ ID NO:106), GQPKAAP (SEQ ID NO: 175), and GGSGSSGSGG (SEQ ID NO:148).

96 . The binding protein of claim 91 , wherein
(a) L 1 comprises the sequence GQPKAAP (SEQ ID NO: 175), L 2 comprises the sequence TKGPS (SEQ ID NO:106), L 3 comprises the amino acid S, and L 4 comprises the sequence RT; (b) L 1 comprises the sequence GGGGSGGGGS (SEQ ID NO:104), L 2 comprises the sequence GGGGSGGGGS (SEQ ID NO:104), L 3 is 0 amino acids in length, and L 4 is 0 amino acids in length; (c) L 1 comprises the sequence GGSGSSGSGG (SEQ ID NO:148), L 2 comprises the sequence GGSGSSGSGG (SEQ ID NO:148), L 3 is 0 amino acids in length, and L 4 is 0 amino acids in length; or (d) L 1 comprises the sequence GGGGSGGGGSGGGGS (SEQ ID NO:105), L 2 is 0 amino acids in length, L 3 comprises the sequence GGGGSGGGGSGGGGS (SEQ ID NO:105), and L 4 is 0 amino acids in length.

97 . The binding protein of claim 91 , wherein the binding protein is capable of inhibiting the function of one or more target proteins.

98 . The binding protein of claim 91 , wherein the binding protein comprises one, two, or three antigen binding sites that specifically bind a target protein selected from the group consisting of A2AR, APRIL, ATPDase, BAFF, BAFFR, BCMA, BlyS, BTK, BTLA, B7DC, B7H1, B7H4, B7H5, B7H6, B7H7, B7RP1, B7-4, C3, C5, CCL2, CCL3, CCL4, CCL5, CCL7, CCL8, CCL11, CCL15, CCL17, CCL19, CCL20, CCL21, CCL24, CCL25, CCL26, CCR3, CCR4, CD3, CD19, CD20, CD23, CD24, CD27, CD28, CD38, CD39, CD40, CD70, CD80, CD86, CD122, CD137, CD137L, CD152, CD154, CD160, CD272, CD273, CD274, CD275, CD276, CD278, CD279, CDH1, chitinase, CLEC9, CLEC91, CRTH2, CSF-1, CSF-2, CSF-3, CX3CL1, CXCL12, CXCL13, CXCR3, DNGR-1, ectonucleoside triphosphate diphosphohydrolase 1, EGFR, ENTPD1, FCER1A, FCER1, FLAP, FOLH1, Gi24, GITR, GITRL, GM-CSF, Her2, HHLA2, HMGB1, HVEM, ICOSLG, IDO, IFNα, IgE, IGF1R, IL2Rbeta, IL1, IL1A, IL1B, IL1F10, IL2, IL4, IL4Ra, IL5, IL5R, IL6, IL7, IL7Ra, IL8, IL9, IL9R, IL10, rhIL10, IL12, IL13, IL13Ra1, IL13Ra2, IL15, IL17, IL17Rb, IL18, IL22, IL23, IL25, IL27, IL33, IL35, ITGB4, ITK, KIR, LAG3, LAMP1, leptin, LPFS2, MHC class II, NCR3LG1, NKG2D, NTPDase-1, OX40, OX40L, PD-1H, platelet receptor, PROM1, S152, SISP1, SLC, SPG64, ST2, STEAP2, Syk kinase, TACI, TDO, T14, TIGIT, TIM3, TLR, TLR2, TLR4, TLR5, TLR9, TMEF1, TNFα, TNFRSF7, Tp55, TREM1, TSLP, TSLPR, TWEAK, VEGF, VISTA, Vstm3, WUCAM, and XCR1.

99 . The binding protein of claim 91 , wherein the C H3 domain of the second polypeptide chain comprises amino acid substitutions at positions corresponding to positions 354 and 366 of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are S354C and T366W; and wherein the C H3 domain of the third polypeptide chain comprises amino acid substitutions at positions corresponding to positions 349, 366, 368, and 407 of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are Y349C, T366S, L368A, and Y407V.

100 . The binding protein of claim 91 , wherein the C H3 domain of the second polypeptide chain comprises amino acid substitutions at positions corresponding to positions 349, 366, 368, and 407 of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are Y349C, T366S, L368A, and Y407V; and wherein the C H3 domain of the third polypeptide chain comprises amino acid substitutions at positions corresponding to positions 354 and 366 of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are S354C and T366W.

101 . The binding protein of claim 91 , wherein the C H3 domains of the second and the third polypeptide chains both comprise amino acid substitutions at positions corresponding to positions 428 and 434 of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are M428L and N434S.

102 . The binding protein of claim 91 , wherein the C H3 domains of the second and the third polypeptide chains are human IgG1 or IgG4 C H3 domains, and wherein only one of the C H3 domains comprises amino acid substitutions at positions corresponding to positions 435 and 436 of human IgG1 or IgG4 according to EU Index, wherein the amino acid substitutions are H435R and Y436F.

103 . The binding protein of claim 91 , wherein the C H3 domains of the second and the third polypeptide chains are human IgG4 C H3 domains, and wherein the C H3 domains each comprise amino acid substitutions at positions corresponding to positions 228 and 409 of human IgG4 according to EU Index, wherein the amino acid substitutions are S228P and R409K.

104 . The binding protein of claim 91 , wherein the C H3 domains of the second and the third polypeptide chains are human IgG4 C H3 domains, and wherein the C H3 domains each comprise amino acid substitutions at positions corresponding to positions 234 and 235 of human IgG4 according to EU Index, wherein the amino acid substitutions are F234A and L235A.

105 . The binding protein of claim 91 , wherein th

Assignees

• Sanofi Sa

Inventors

• Yang Zhi-Yong
• Nabel Gary J
• Wu Lan
• Seung Edward
• Wei Ronnie
• Beninga Jochen
• Rao Ercole
• Leuschner Wulf Dirk
• Beil Christian
• Lange Christian
• Corvey Carsten

Classifications

• C07K16/2818 Primary

  against CD28 or CD152 · CPC title

• C07K2317/75

  Agonist effect on antigen · CPC title

• C07K16/247

  IL-4 · CPC title

• C07K2319/00

  Fusion polypeptide · CPC title

• C07K16/32

  against translation products of oncogenes · CPC title