BCMA NK cell engager therapy for autoimmune disorders

Abstract

The present disclosure relates to a method of treating a B-cell mediated autoimmune disorder in a subject comprising administering to the subject: a binding protein comprising a first antigen binding domain (ABD) with binding specificity to BCMA and a second ABD with binding specificity to NKp46.

Description

[0001] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP BCMA NK CELL ENGAGER THERAPY FOR AUTOIMMUNE DISORDERS CROSS REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to European Application No. 25305436.5, filed March 26, 2025, the contents of which is incorporated in its entirety.

[0003] BACKGROUND

[0004] Autoimmune diseases occur when the immune system mistakenly attacks the body's own tissues, leading to inflammation and damage. This can involve various immune cells, including autoreactive B cells, which produce antibodies that target self-antigens. Plasma cells are the primary source of pathogenic autoantibodies, and the expression of BCMA on plasma cells and plasmablasts makes BCMA a promising target for B-cell mediated autoimmune diseases. While immunosuppressive treatments such as corticosteroids, diseasemodifying antirheumatic drugs (DMARDs), and biologic therapies may be effective in controlling symptoms, reducing inflammation, and preventing disease progression in autoimmune diseases, not all patients respond to these treatments, and some may experience significant side effects. Thus, there exists a need in the art for potent therapies in treating autoimmune diseases.

[0005] SUMMARY

[0006] The present disclosure relates to a method of treating a B-cell mediated autoimmune disorder in a subject, comprising administering to the subject: a binding protein comprising a first antigen binding domain (ABD) with binding specificity to BCMA and a second ABD with binding specificity to NKp46; and thereby treating the B-cell mediated autoimmune disorder in the subject.

[0007] In one aspect, the disclosure provides a method of treating a B-cell mediated autoimmune disorder in a subject, comprising administering to the subject a binding protein comprising a first antigen binding domain (ABD) with binding specificity to BCMA and a second ABD with binding specificity to NKp46, thereby treating the B-cell mediated autoimmune disorder in the subject.Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP In certain embodiments, (a) the first ABD comprises: (a1) a first immunoglobulin heavy chain variable domain (VH1) comprising an HCDR1 sequence comprising the amino acid sequence of GFTFSNFGMH (SEQ ID NO: 1), an HCDR2 sequence comprising the amino acid sequence of VIWSDETNR (SEQ ID NO: 2), and an HCDR3 sequence comprising the amino acid sequence of DQQYCSSDSCFTWFDP (SEQ ID NO: 3); and (a2) a first immunoglobulin light chain variable domain (VL1) comprising an LCDR1 sequence comprising the amino acid sequence of CX1SSTGX2VTPX3X4YAN (SEQ ID NO: 4), wherein X1is R or A, X2is T or A, X3is S or G, and X4is N or Y, an LCDR2 sequence comprising the amino acid sequence of DNNX5X6PP (SEQ ID NO: 5), wherein X5is S, I, or N and X6is R or K, and an LCDR3 sequence comprising the amino acid sequence of ALX7X8GX9QWV (SEQ ID NO: 6), wherein X7is W or Y, X8is F or Y, and X9is N or G; and (b) the second ABD comprises binding specificity to NKp46.

[0008] In certain embodiments, (b) the second ABD comprises: (b1) a second immunoglobulin heavy chain variable domain (VH2) comprising: - an HCDR1 sequence comprising DYVIN, an HCDR2 sequence comprising EIYPGSGTNYYNEKFKA, and an HCDR3 sequence comprising RGRYGLYAMDY; - an HCDR1 sequence comprising GYTFSDYVIN (SEQ ID NO: 19), an HCDR2 sequence comprising EIYPGSGTN (SEQ ID NO: 20), and an HCDR3 sequence comprising RGRYGLYAMDY (SEQ ID NO: 21); - an HCDR1 sequence comprising SDYAWN (SEQ ID NO: 22), an HCDR2 sequence comprising YITYSGSTSYNPSLES (SEQ ID NO: 23), and an HCDR3 sequence comprising GGYYGSSWGVFAY (SEQ ID NO: 24); - an HCDR1 sequence comprising EYTMH (SEQ ID NO: 25), an HCDR2 sequence comprising GISPNIGGTSYNQKFKG (SEQ ID NO: 26), and an HCDR3 sequence comprising RGGSFDY (SEQ ID NO: 27); -an HCDR1 sequence comprising SFTMH (SEQ ID NO: 28), an HCDR2 sequence comprising YINPSSGYTEYNQKFKD (SEQ ID NO: 29), and an HCDR3 sequence comprising GSSRGFDY (SEQ ID NO: 30); or - an HCDR1 sequence comprising SDYAWN (SEQ ID NO: 31), an HCDR2 sequence comprising YITYSGSTNYNPSLKS (SEQ ID NO: 32), and an HCDR3 sequence comprising CWDYALYAMDC (SEQ ID NO: 33); and (b2) a second immunoglobulin light chain variable domain (VL2) comprising: - an LCDR1 sequence comprisingAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP RASQDISNYLN (SEQ ID NO: 34), an LCDR2 sequence comprising YTSRLHS (SEQ ID NO: 35), and an LCDR3 sequence comprising QQGNTRPWT (SEQ ID NO: 36); - an LCDR1 sequence comprising RVSENIYSYLA (SEQ ID NO: 37), an LCDR2 sequence comprising NAKTLAE (SEQ ID NO: 38), and an LCDR3 sequence comprising QHHYGTPWT (SEQ ID NO: 39); - an LCDR1 sequence comprising RASQSISDYLH (SEQ ID NO: 40), an LCDR2 sequence comprising YASQSIS (SEQ ID NO: 41), and an LCDR3 sequence comprising QNGHSFPLT (SEQ ID NO: 42); - an LCDR1 sequence comprising RASENIYSNLA (SEQ ID NO: 43), an LCDR2 sequence comprising AATNLAD (SEQ ID NO: 44), and an LCDR3 sequence comprising QHFWGTPRT (SEQ ID NO: 45); or - an LCDR1 sequence comprising RTSENIYSYLA (SEQ ID NO: 46), an LCDR2 sequence comprising NAKTLAE (SEQ ID NO: 47), and an LCDR3 sequence comprising QHHYDTPLT (SEQ ID NO: 48).

[0009] In certain embodiments, the VL1 comprises: - an LCDR1 sequence comprising the amino acid sequence of CASSTGTVTPSNYAN (SEQ ID NO: 7), an LCDR2 sequence comprising the amino acid sequence of DNNSRPP (SEQ ID NO: 8), and an LCDR3 sequence comprising the amino acid sequence of ALWFGNQWV (SEQ ID NO: 9); - an LCDR1 sequence comprising the amino acid sequence of CRSSTGTVTPSNYAN (SEQ ID NO: 10), an LCDR2 sequence comprising the amino acid sequence of DNNSRPP (SEQ ID NO: 11), and an LCDR3 sequence comprising the amino acid sequence of ALWFGNQWV (SEQ ID NO: 12); - an LCDR1 sequence comprising the amino acid sequence of CASSTGAVTPSNYAN (SEQ ID NO: 13), an LCDR2 sequence comprising the amino acid sequence of DNNIKPP (SEQ ID NO: 14), and an LCDR3 sequence comprising the amino acid sequence of ALWYGGQWV (SEQ ID NO: 15); or - an LCDR1 sequence comprising the amino acid sequence of CASSTGAVTPGYYAN (SEQ ID NO: 16), an LCDR2 sequence comprising the amino acid sequence of DNNNKPP (SEQ ID NO: 17), and an LCDR3 sequence comprising the amino acid sequence of ALYYGGQWV (SEQ ID NO: 18).

[0010] In certain embodiments, (a) the first ABD comprises: (a1) a first immunoglobulin heavy chain variable domain (VH1) comprising an HCDR1 sequence comprising the amino acid sequence of GFTFSNFG (SEQ ID NO: 112),Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP an HCDR2 sequence comprising the amino acid sequence of IWSDETNR (SEQ ID NO: 113), and an HCDR3 sequence comprising the amino acid sequence of ARDQQYCSSDSCFTWFDP (SEQ ID NO: 114); and (a2) a first immunoglobulin light chain variable domain (VL1) comprising an LCDR1 sequence comprising the amino acid sequence of TGTVTPSNY (SEQ ID NO: 118), an LCDR2 sequence comprising the amino acid sequence of DNN (SEQ ID NO: 119), and an LCDR3 sequence comprising the amino acid sequence of ALWFGNQWV (SEQ ID NO: 120).

[0011] In certain embodiments, (a) the first ABD comprises: (a1) a first immunoglobulin heavy chain variable domain (VH1) comprising an HCDR1 sequence comprising the amino acid sequence of NFGMH (SEQ ID NO: 115), an HCDR2 sequence comprising the amino acid sequence of VIWSDETNRYYADSVKG (SEQ ID NO: 116), and an HCDR3 sequence comprising the amino acid sequence of DQQYCSSDSCFTWFDP (SEQ ID NO: 117); and (a2) a first immunoglobulin light chain variable domain (VL1) comprising an LCDR1 sequence comprising the amino acid sequence of ASSTGTVTPSNYAN (SEQ ID NO: 121), an LCDR2 sequence comprising the amino acid sequence of DNNSRPP (SEQ ID NO: 122), and an LCDR3 sequence comprising the amino acid sequence of ALWFGNQWV (SEQ ID NO: 123).

[0012] In certain embodiments, - the VH1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 55; - the VH1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 50; - the VH1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 51 ; - the VH1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence that is at least about 90% identical to the aminoAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP acid sequence of SEQ ID NO: 52; - the VH1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 53; or - the VH1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 54.

[0013] In certain embodiments, - the VH1 comprises an amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence of SEQ ID NO: 55; - the VH1 comprises an amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence of SEQ ID NO: 50; - the VH1 comprises an amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence of SEQ ID NO: 51; - the VH1 comprises an amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence of SEQ ID NO: 52; - the VH1 comprises an amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence of SEQ ID NO: 53; or - the VH1 comprises an amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence of SEQ ID NO: 54.

[0014] In certain embodiments, - the VH2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 56, and wherein the VL2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 64; - the VH2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 57, and wherein the VL2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 65; - the VH2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 58, and wherein the VL2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 66; - the VH2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 59, and wherein the VL2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 67; - the VH2 comprises an amino acid sequenceAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 60, and wherein the VL2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 68; - the VH2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 61 , and wherein the VL2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 69; - the VH2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 62, and wherein the VL2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 70; or - the VH2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 63, and wherein the VL2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 71.

[0015] In certain embodiments, - the VH2 comprises an amino acid sequence of SEQ ID NO: 56, and wherein the VL2 comprises an amino acid sequence of SEQ ID NO: 64; - the VH2 comprises an amino acid sequence of SEQ ID NO: 57, and wherein the VL2 comprises an amino acid sequence of SEQ ID NO: 65; - the VH2 comprises an amino acid sequence of SEQ ID NO: 58, and wherein the VL2 comprises an amino acid sequence of SEQ ID NO: 66; - the VH2 comprises an amino acid sequence of SEQ ID NO: 59, and wherein the VL2 comprises an amino acid sequence of SEQ ID NO: 67; - the VH2 comprises an amino acid sequence of SEQ ID NO: 60, and wherein the VL2 comprises an amino acid sequence of SEQ ID NO: 68; - the VH2 comprises an amino acid sequence of SEQ ID NO: 61, and wherein the VL2 comprises an amino acid sequence of SEQ ID NO: 69; - the VH2 comprises an amino acid sequence of SEQ ID NO: 62, and wherein the VL2 comprises an amino acid sequence of SEQ ID NO: 70; or - the VH2 comprises an amino acid sequence of SEQ ID NO: 63, and wherein the VL2 comprises an amino acid sequence of SEQ ID NO: 71.

[0016] In certain embodiments, the binding protein further comprises all or part of an immunoglobulin Fc domain or variant thereof. In certain embodiments, the Fc domain is an lgG1 Fc domain. In certain embodiments, the lgG1 Fc domain is a human IgG 1 Fc domain. In certain embodiments, all or part of the immunoglobulinAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP Fc domain or variant thereof binds to a human Fc-y receptor. In certain embodiments, all or part of the immunoglobulin Fc domain or variant thereof binds to a human CD16A (FcyRIII) polypeptide.

[0017] In certain embodiments, the Fc domain comprises a native glycan at amino acid position 297, according to Ell numbering.

[0018] In certain embodiments, the binding protein is N-glycosylated. In certain embodiments, the Fc domain or variant thereof comprises a first Fc heavy chain and a second Fc heavy chain.

[0019] In certain embodiments, at least one Fc heavy chain comprises an engineered intrachain disulfide bond mediated by a pair of cysteines (C) that substitute for: (i) a leucine (L) at amino acid position 242 and a lysine (K) at amino acid position 334; or (ii) an arginine (R) at amino acid position 292 and a valine (V) at amino acid position 302; wherein the amino acid positions are according to Ell numbering.

[0020] In certain embodiments, the first Fc heavy chain or the second Fc heavy chain comprises the pair of cysteines. In certain embodiments, the first and the second Fc heavy chain each comprise the pair of cysteines. In certain embodiments, the first and the second Fc heavy chain each comprise the L242C I K334C substitutions. In certain embodiments, the first and the second Fc heavy chain each comprise the R292C I V302C substitutions.

[0021] In certain embodiments, at least one Fc heavy chain comprises a substitution at amino acid position 332, according to Ell numbering. In certain embodiments, the substitution at amino acid position 332 is a glutamic acid (E).

[0022] In certain embodiments, at least one Fc heavy chain further comprises one or more substitutions at amino acid positions 236, 239, or 330, according to Ell numbering. In certain embodiments, the substitution at amino acid position 236 is an alanine (A). In certain embodiments, the substitution at amino acid position 239 is an aspartic acid (D). In certain embodiments, the substitution at amino acid position 330 is a leucine (L).

[0023] In certain embodiments, at least one Fc heavy chain further comprises an aspartic acid (D) at amino acid position 239, and a glutamic acid (E) at amino acid position 332, according to Ell numbering. In certain embodiments, at least oneAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP Fc heavy chain further comprises an alanine (A) at amino acid position 236, an aspartic acid (D) at amino acid position 239, and a glutamic acid (E) at amino acid position 332, according to Ell numbering. In certain embodiments, at least one Fc heavy chain further comprises an alanine (A) at amino acid position 236, an aspartic acid (D) at amino acid position 239, a leucine (L) at amino acid position 330, and a glutamic acid (E) at amino acid position 332, according to Ell numbering.

[0024] In certain embodiments, the binding protein comprises at least two polypeptide chains that form at least two antigen-binding sites, wherein at least one polypeptide chain comprises a structure represented by the formula:

[0025] VL1-L1-VL2-L2-CL [I];

[0026] and at least one polypeptide chain comprises a structure represented by the formula:

[0027] VH2-L3-VH1-L4-CH1 [II];

[0028] wherein:

[0029] CL is an immunoglobulin light chain constant domain;

[0030] CH1 is an immunoglobulin CH1 heavy chain constant domain; and L1, L2, L3, and L4 are amino acid linkers, wherein any one or more of L1 , L2, L3, and L4 are optionally absent, and

[0031] wherein the polypeptides of formula I and the polypeptides of formula II form a cross-over light chain-heavy chain pair.

[0032] In certain embodiments, the binding protein comprises three polypeptide chains that form two antigen-binding sites, wherein one polypeptide chain comprises a structure represented by the formula:

[0033] VL1-L1-VL2-L2-CL [I];

[0034] one polypeptide chain comprises a structure represented by the formula: VH2-L3-VH1-L4-CH1-hinge-CH2-CH3 [III]; and

[0035] one polypeptide chain comprises a structure represented by the formula: hinge-CH2-CH3 [IV]

[0036] wherein:

[0037] CL is an immunoglobulin light chain constant domain;

[0038] CH1 is an immunoglobulin CH1 heavy chain constant domain; CH2 is an immunoglobulin CH2 heavy chain constant domain;Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP CH3 is an immunoglobulin CH3 heavy chain constant domain; hinge is an immunoglobulin hinge region connecting the CH1 and CH2 domains;

[0039] and

[0040] L1, L2, L3, and L4 are amino acid linkers, wherein any one or more of L1 , L2, L3, and L4 are optionally absent, and

[0041] wherein the polypeptides of formula I and the polypeptides of formula II form a cross-over light chain-heavy chain pair.

[0042] In certain embodiments: (a) L1, L2, L3, and L4 each independently are zero amino acids in length or comprise a sequence selected from the group consisting of GGGGSGGGGS, GGGGSGGGGSGGGGS (SEQ ID NO: 82), S, RT, TKGPS (SEQ ID NO: 83), GQPKAAP (SEQ ID NO: 84), and GGSGSSGSGG (SEQ ID NO: 85); or (b) L1, L2, L3, and L4 each independently comprise a sequence selected from the group consisting of GGGGSGGGGS, GGGGSGGGGSGGGGS, S, RT, TKGPS, GQPKAAP, and GGSGSSGSGG.

[0043] In certain embodiments, L1 and L2 each comprise the amino acid sequence GGGGSGGGGS. In certain embodiments, L3 and L4 are each absent.

[0044] In certain embodiments, (a) the first ABD of the binding protein comprises a first immunoglobulin heavy chain variable domain (VH1 ) comprising an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 49, and a first immunoglobulin light chain variable domain (VL1) comprising an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 55; and (b) the second ABD of the binding protein comprises a second immunoglobulin heavy chain variable domain (VH2) comprising an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 56, and a second immunoglobulin light chain variable domain (VL2) comprising an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 64.

[0045] In certain embodiments, the binding protein comprises: (i) a first polypeptide chain comprising an amino acid sequence of SEQ ID NO: 72; (ii) a second polypeptide chain comprising an amino acid sequence of SEQ ID NO: 73;Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP and (iii) a third polypeptide chain comprising an amino acid sequence of SEQ ID NO: 74.

[0046] In certain embodiments, (b) the second ABD comprises: (b1) a second immunoglobulin heavy chain variable domain (VH2) comprising an HCDR1 sequence comprising the amino acid sequence of GYTFSDYV (SEQ ID NO: 87), an HCDR2 sequence comprising the amino acid sequence of IYPGSGTN (SEQ ID NO: 90), and an HCDR3 sequence comprising the amino acid sequence of ARRGRYGLYAMDY (SEQ ID NO: 93); and (b2) a second immunoglobulin light chain variable domain (VL1) comprising an LCDR1 sequence comprising the amino acid sequence of QDISNY (SEQ ID NO: 96), an LCDR2 sequence comprising the amino acid sequence of YTS (SEQ ID NO: 99), and an LCDR3 sequence comprising the amino acid sequence of QQGNTRPWT (SEQ ID NO: 111).

[0047] In certain embodiments, (b) the second ABD comprises: (b1) a second immunoglobulin heavy chain variable domain (VH2) comprising an HCDR1 sequence comprising the amino acid sequence of DYVIN (SEQ ID NO: 80), an HCDR2 sequence comprising the amino acid sequence of EIYPGSGTNYYNEKFKA (SEQ ID NO: 81), and an HCDR3 sequence comprising the amino acid sequence of RGRYGLYAMDY (SEQ ID NO: 21 ); and (b2) a second immunoglobulin light chain variable domain (VL1) comprising an LCDR1 sequence comprising the amino acid sequence of RASQDISNYLN (SEQ ID NO: 34), an LCDR2 sequence comprising the amino acid sequence of YTSRLHS (SEQ ID NO: 35), and an LCDR3 sequence comprising the amino acid sequence of QQGNTRPWT (SEQ ID NO: 36).

[0048] In certain embodiments, the autoimmune disorder is a B-cell mediated autoimmune disorder.

[0049] In certain embodiments, the autoimmune disorder is a plasma cell mediated autoimmune disorder.

[0050] In certain embodiments, the autoimmune disorder is lupus.

[0051] In certain embodiments, the lupus is systemic lupus erythematosus (SLE).

[0052] In certain embodiments, the autoimmune disorder is rheumatoid arthritis (RA).Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP In certain embodiments, the subject has a reduced serum immunoglobulin level compared to a subject that was not administered the binding protein.

[0053] In certain embodiments, the subject has a reduced autoantibody level compared to a subject that was not administered the binding protein.

[0054] In certain embodiments, the serum immunoglobulin and / or autoantibody is IgA, IgM and / or IgG.

[0055] In certain embodiments, the subject has a reduced serum rheumatoid factor (RF) level compared to a subject that was not administered the binding protein.

[0056] In certain embodiments, the binding protein is administered intravenously or subcutaneously.

[0057] BRIEF DESCRIPTION OF THE DRAWINGS

[0058] The foregoing and other features and advantages of the present disclosure will be more fully understood from the following detailed description of illustrative embodiments taken in conjunction with the accompanying drawings.

[0059] FIG. 1 depicts a distinct NKp46-BCMA NKCE format that engages BCMA at the surface of a cell while also recruiting NK cells through the dual engagement of both NKp46 and a Fey receptor, CD16a, inducing ADCC activity with an enhanced Fc competent format. The enhanced Fc competent format (hereinafter, termed “CODV-OL1-ADE-DSB”) includes: (1) ADE mutations (G236A / S239D / I332E) in CH2 to enhance ADCC-activity; (2) DSB (R292CA / 302C) in CH2 for thermal stabilization and productivity (e.g., expressability in cells and overall yield) ; (3) knob-into-hole mutations (KIH) in CH3 to favor heterodimer formation in Fc - (3a) knob (in heavy chain containing the VH / VL domains): S354C I T366W and (3b) hole (in heavy chain lacking the VH / VL domains): Y349C I T366S / L368A / Y407V; and (4) RF mutations (H435R / Y436F) in one CH3 to favor purification of heterodimers in Fc. All Fc domain amino acid numbering is according to EU. This NKp46-BCMA NKCE_Fc CODV-OL1-ADE-DSB additionally contains two linkers GGGGSGGGGS in the light chain: one between VL anti-BCMA and VLAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP anti-NKp46 and between VL NKp46 and CL. NKp46 binding site: 3D9; BCMA binding site: CA10v7; CD16 binding through ADCC-competent Fc domain.

[0060] DETAILED DESCRIPTION

[0061] The disclosure provides methods of treating B-cell mediated autoimmune disorders (e.g., lupus or SLE) with binding proteins that bind one surface biomarker on immune NK cells i.e. , NKp46 and one antigen of interest on the cell membrane of normal and malignant plasma cells i.e., BCMA, and is capable of redirecting NK cells to lyse a target cell that expresses the BCMA surface biomarker.

[0062] It is to be understood that the methods described in this disclosure are not limited to particular methods and experimental conditions disclosed herein as such methods and conditions may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0063] Furthermore, the experiments described herein, unless otherwise indicated, use conventional molecular and cellular biological and immunological techniques within the skill of the art. Such techniques are well known to the skilled worker and are explained fully in the literature. See, e.g., Ausubel, etal., ed., Current Protocols in Molecular Biology, John Wiley & Sons, Inc., NY, N.Y. (1987-2008), including all supplements, Molecular Cloning: A Laboratory Manual (Fourth Edition) by MR Green and J. Sambrook and Harlow etal., Antibodies: A Laboratory Manual, Chapter 14, Cold Spring Harbor Laboratory, Cold Spring Harbor (2013, 2ndedition).

[0064] Unless otherwise defined, scientific and technical terms used herein have the meanings that are commonly understood by those of ordinary skill in the art. In the event of any latent ambiguity, definitions provided herein take precedent over any dictionary or extrinsic definition. Unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular. The use of “or” means “and / or” unless stated otherwise. The use of the term “including,” as well as other forms, such as “includes” and “included,” is not limiting.

[0065] Generally, nomenclature used in connection with cell and tissue culture, molecular biology, immunology, microbiology, genetics and protein and nucleic acid chemistry and hybridization described herein is well-known and commonly used inAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP the art. The methods and techniques provided herein are generally performed according to conventional methods well known in the art and as described in various general and more specific references that are cited and discussed throughout the present specification unless otherwise indicated. Enzymatic reactions and purification techniques are performed according to manufacturer’s specifications, as commonly accomplished in the art or as described herein. The nomenclatures used in connection with, and the laboratory procedures and techniques of, analytical chemistry, synthetic organic chemistry, and medicinal and pharmaceutical chemistry described herein are those well-known and commonly used in the art. Standard techniques are used for chemical syntheses, chemical analyses, pharmaceutical preparation, formulation, and delivery, and treatment of patients.

[0066] That the disclosure may be more readily understood, select terms are defined below.

[0067] The term “polypeptide” refers to any polymeric chain of amino acids and encompasses native or artificial proteins, polypeptide analogs or variants of a protein sequence, or fragments thereof, unless otherwise contradicted by context. A polypeptide may be monomeric or polymeric. A polypeptide fragment comprises at least about 5 contiguous amino acids, at least about 10 contiguous amino acids, at least about 15 contiguous amino acids, or at least about 20 contiguous amino acids, for example.

[0068] The term “isolated protein” or “isolated polypeptide” refers to a protein or polypeptide that by virtue of its origin or source of derivation is not associated with naturally associated components that accompany it in its native state; is substantially free of other proteins from the same species; is expressed by a cell from a different species; or does not occur in nature. Thus, a protein or polypeptide that is chemically synthesized or synthesized in a cellular system different from the cell from which it naturally originates will be “isolated” from its naturally associated components. A protein or polypeptide may also be rendered substantially free of naturally associated components by isolation using protein purification techniques well known in the art.

[0069] As used herein, the term “binding protein” or “binding polypeptide” shall refer to a protein or polypeptide (e.g., an antibody or immunoadhesin) that containsAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP at least one binding site which is responsible for selectively binding to a target antigen of interest (e.g., a human target antigen). Exemplary binding sites include an antibody variable domain, a ligand binding site of a receptor, or a receptor binding site of a ligand. In certain aspects, the binding proteins or binding polypeptides comprise multiple (e.g., two, three, four, or more) binding sites. In certain aspects, the binding protein or binding polypeptide is a therapeutic enzyme.

[0070] The term “ligand” refers to any substance capable of binding, or of being bound, to another substance. Similarly, the term “antigen” refers to any substance to which an antibody may be generated. Although “antigen” is commonly used in reference to an antibody binding substrate, and “ligand” is often used when referring to receptor binding substrates, these terms are not distinguishing, one from the other, and encompass a wide range of overlapping chemical entities. For the avoidance of doubt, antigen and ligand are used interchangeably throughout herein. Antigens / ligands may be a peptide, a polypeptide, a protein, an aptamer, a polysaccharide, a sugar molecule, a carbohydrate, a lipid, an oligonucleotide, a polynucleotide, a synthetic molecule, an inorganic molecule, an organic molecule, and any combination thereof.

[0071] The dissociation constant (KD) of a binding protein can be determined, for example, by surface plasmon resonance (SPR). Generally, SPR analysis measures real-time binding interactions between ligand (a target antigen on a biosensor matrix) and analyte (a binding protein in solution) by SPR using the BIAcore system (Pharmacia Biosensor; Piscataway, NJ). Surface plasmon analysis can also be performed by immobilizing the analyte (binding protein on a biosensor matrix) and presenting the ligand (target antigen). The term "KD” as used herein refers to the dissociation constant of the interaction between a particular binding protein and a target antigen.

[0072] The term “specifically binds” as used herein, refers to the ability of an antibody or an immunoadhesin to bind to a target (e.g., an antigen) with a dissociation constant (KD) of at most about 1 x 10’6M, about 1 x 10’7M, about 1 x 10’8M, about 1 x 10’9M, about 1 x 1O’10M, about 1 x 10’11M, about 1 x 10’12M or less, and / or to bind to an antigen with an affinity that is at least about two-fold greater than its affinity for a nonspecific antigen. Specific binding of an antibody can be to aAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP target antigen through the complementary determining region (CDR) sequences. An antibody can also specifically bind to FcRs, such as FcRn or FcyRllla through the Fc region.

[0073] As used herein, the term “antibody” refers to such assemblies (e.g., intact antibody molecules, immunoadhesins, or variants thereof) which have significant known specific immunoreactive activity to an antigen of interest (e.g. a tumor associated antigen). Antibodies and immunoglobulins comprise light and heavy chains, with or without an interchain covalent linkage between them. Basic immunoglobulin structures in vertebrate systems are relatively well understood.

[0074] As will be discussed in more detail below, the generic term “antibody” comprises five distinct classes of antibody that can be distinguished biochemically. While all five classes of antibodies are clearly within the scope of the current disclosure, the following discussion will generally be directed to the IgG class of immunoglobulin molecules. With regard to IgG, immunoglobulins comprise two identical light chains of molecular weight approximately 23,000 Daltons, and two identical heavy chains of molecular weight 53,000-70,000. The four chains are joined by disulfide bonds in a “Y” configuration wherein the light chains bracket the heavy chains starting at the mouth of the “Y” and continuing through the variable region.

[0075] Light chains of immunoglobulin are classified as either kappa (K) or lambda (A). Each heavy chain class may be bound with either a kappa or lambda light chain. In general, the light and heavy chains are covalently bonded to each other, and the "tail" portions of the two heavy chains are bonded to each other by covalent disulfide linkages or non-covalent linkages when the immunoglobulins are generated either by hybridomas, B cells, or genetically engineered host cells. In the heavy chain, the amino acid sequences run from an N-terminus at the forked ends of the Y configuration to the C-terminus at the bottom of each chain. Those skilled in the art will appreciate that heavy chains are classified as gamma (y), mu (p), alpha (a), delta (5), or epsilon (E), with some subclasses among them (e.g., yl-y4). It is the nature of this chain that determines the “class” of the antibody as IgG, IgM, IgA, or IgE, respectively. The immunoglobulin isotype subclasses (e.g., lgG1, lgG2, lgG3, lgG-4, lgA1, etc.) are well-characterized and are known to confer functionalAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP specialization. Modified versions of each of these classes and isotypes are readily discernable to the skilled artisan in view of the instant disclosure and, accordingly, are within the scope of the current disclosure.

[0076] Both the light and heavy chains are divided into regions of structural and functional homology. The term “region” refers to a part or portion of an immunoglobulin or antibody chain and includes constant region or variable regions, as well as more discrete parts or portions of said regions. For example, light chain variable regions include “complementarity determining regions” or “CDRs” interspersed among “framework regions” or “FRs,” as defined herein.

[0077] The regions of an immunoglobulin heavy or light chain may be defined as “constant” (C) region or “variable” (V) regions, based on a relative lack of sequence variation within the regions of various class members in the case of a “constant region,” or based on a significant variation within the regions of various class members in the case of a “variable regions.” The terms “constant region” and “variable region” may also be used functionally. In this regard, it will be appreciated that the variable regions of an immunoglobulin or antibody determine antigen recognition and specificity. Conversely, the constant regions of an immunoglobulin or antibody confer important effector functions such as secretion, trans-placental mobility, Fc receptor binding, complement binding, and the like. The subunit structures and three-dimensional configurations of the constant regions of the various immunoglobulin classes are well-known.

[0078] The constant and variable regions of immunoglobulin heavy and light chains are folded into domains. The term “domain” refers to a globular region of a heavy or light chain comprising peptide loops (e.g., comprising 3 to 4 peptide loops) stabilized, for example, by [3-pleated sheet and / or an intra-chain disulfide bond. Constant region domains on the light chain of an immunoglobulin are referred to interchangeably as “light chain constant region domains,” “CL regions,” “CL domains” or “CK domains.” Constant domains on the heavy chain (e.g., hinge, CH1 , CH2 or CH3 domains) are referred to interchangeably as “heavy chain constant region domains,” “CH” region domains or “CH domains.” Variable domains on the light chain are referred to interchangeably as “light chain variable region domains,” “VL region domains” or “VL domains.” Variable domains on the heavy chain areAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP referred to interchangeably as “heavy chain variable region domains,” “VH region domains” or “VH domains.”

[0079] By convention, the numbering of the amino acids of the variable constant region domains increases as they become more distal from the antigen-binding site or amino-terminus of the immunoglobulin or antibody. The N-terminus of each heavy and light immunoglobulin chain is a variable region and the C-terminus is a constant region. The CH3 and CL domains comprise the carboxy-terminus of the heavy and light chain, respectively. Accordingly, the domains of a light chain immunoglobulin are arranged in a VL-CL orientation, while the domains of the heavy chain are arranged in the VH-CH1-hinge-CH2-CH3 orientation.

[0080] The assignment of amino acids to each variable region domain is in accordance with the definitions of Kabat, Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, MD, 1987 and 1991). Kabat also provides a widely used numbering convention (Kabat numbering) in which corresponding residues between different heavy chain variable regions or between different light chain variable regions are assigned the same number. CDRs 1 , 2 and 3 of a VL domain are also referred to herein, respectively, as CDR-L1 , CDR-L2 and CDR-L3. CDRs 1 , 2 and 3 of a VH domain are also referred to herein, respectively, as CDR-H1 , CDR- H2 and CDR-H3. If so noted, the assignment of CDRs can be in accordance with IMGT® (Lefranc et al., Developmental & Comparative Immunology 27:55-77; 2003) in lieu of Kabat. Numbering of the heavy chain constant region is via the Ell index as set forth in Kabat (Kabat, Sequences of Proteins of Immunological Interest, National Institutes of Health, Bethesda, MD, 1987 and 1991). The precise amino acid sequence boundaries of a given CDR or FR can be readily determined using any of a number of well-known schemes, including those described by Kabat et al. (1991 ), “Sequences of Proteins of Immunological Interest,” 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (“Kabat” numbering scheme), Al-Lazikani et al., (1997) JMB 273, 927-948 (“Chothia” numbering scheme), MacCallum et al., J. Mol. Biol. 262:732-745 (1996), “Antibodyantigen interactions: Contact analysis and binding site topography,” J. Mol. Biol.

[0081] 262, 732-745. (“Contact” numbering scheme), Lefranc M P et al., “IMGT unique numbering for immunoglobulin and T cell receptor variable domains and IgAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP superfamily V-like domains,” Dev Comp Immunol, 2003 January; 27(1):55-77 (“IMGT” numbering scheme), and Honegger A and Pluckthun A, “Yet another numbering scheme for immunoglobulin variable domains: an automatic modeling and analysis tool,” J Mol Biol, 2001 Jun. 8; 309(3):657-70, (AHo numbering scheme).

[0082] The boundaries of a given CDR or FR may vary depending on the scheme used for identification. For example, the Kabat scheme is based on structural alignments, while the Chothia scheme is based on structural information. Numbering for both the Kabat and Chothia schemes is based upon the most common antibody region sequence lengths, with insertions accommodated by insertion letters, for example, “30a,” and deletions appearing in some antibodies. The two schemes place certain insertions and deletions (“indels”) at different positions, resulting in differential numbering. The Contact scheme is based on analysis of complex crystal structures and is similar in many respects to the Chothia numbering scheme.

[0083] As used herein, the CDRs of an antibody can be determined according to the numbering system called, “IMGT” described in Lefranc (1999), The Immunologist, vol. 7:132-136 and Lefranc et al. (1999), Nucleic Acids Res., vol. 27:209-212.

[0084] As used herein, the CDRs of an antibody can be determined according to the Chothia numbering scheme, which refers to the location of immunoglobulin structural loops. Chothia and Lesk (1987), J. Mol. Biol., vol.196:901-917; Al-Lazikani et al. (1997), J. Mol. Biol., vol. 273:927-948; Chothia et al. (1992), J. Mol. Biol., vol.

[0085] 227:799-817; Tramontane A et al. (1990), J. Mol. Biol. vol. 215(1): 175-82.

[0086] As used herein, the CDRs of an antibody can be determined according to the Honegger-Pluckthun numbering scheme described in Honnegger and Pluckthun (2001), J. Mol. Biol., vol. 309(3):657-670.

[0087] As used herein, the term “VH domain” includes the amino terminal variable domain of an immunoglobulin heavy chain, and the term “VL domain” includes the amino terminal variable domain of an immunoglobulin light chain.

[0088] As used herein, the term “CH1 domain” includes the first (most amino terminal) constant region domain of an immunoglobulin heavy chain that extends, e.g., from about positions 114-223 in the Kabat numbering system (Ell positionsAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP 118-215). The CH1 domain is adjacent to the VH domain and amino terminal to the hinge region of an immunoglobulin heavy chain molecule and does not form a part of the Fc region of an immunoglobulin heavy chain.

[0089] As used herein, the term “hinge region” includes the portion of a heavy chain molecule that joins the CH1 domain to the CH2 domain. The hinge region comprises approximately 25 residues and is flexible, thus allowing the two N-terminal antigen binding regions to move independently. Hinge regions can be subdivided into three distinct domains: upper, middle, and lower hinge domains (Roux et al. J. Immunol. 1998, 161 :4083).

[0090] As used herein, the term “CH2 domain” includes the portion of a heavy chain immunoglobulin molecule that extends, e.g., from about positions 244-360 in the Kabat numbering system (Ell positions 231-340). The CH2 domain is unique in that it is not closely paired with another domain. Rather, two N-linked branched carbohydrate chains are interposed between the two CH2 domains of an intact native IgG molecule. In one embodiment, a binding polypeptide of the current disclosure comprises a CH2 domain derived from an lgG1 molecule (e.g. a human lgG1 molecule).

[0091] As used herein, the term “CH3 domain” includes the portion of a heavy chain immunoglobulin molecule that extends approximately 110 residues from N-terminus of the CH2 domain, e.g., from about positions 361-476 of the Kabat numbering system (Ell positions 341-445). The CH3 domain typically forms the C-terminal portion of the antibody. In some immunoglobulins, however, additional domains may extend from the CH3 domain to form the C-terminal portion of the molecule (e.g., the CH4 domain in the p chain of IgM and the e chain of IgE). In one embodiment, a binding polypeptide of the current disclosure comprises a CH3 domain derived from an lgG1 molecule (e.g., a human lgG1 molecule).

[0092] As used herein, the term “CL domain” includes the constant region domain of an immunoglobulin light chain that extends, e.g., from about Kabat position 107A to about Kabat position 216. The CL domain is adjacent to the VL domain. In one embodiment, a binding polypeptide of the current disclosure comprises a CL domain derived from a kappa light chain (e.g., a human kappa light chain).Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP The variable regions of an antibody allow it to selectively recognize and specifically bind epitopes on antigens. That is, the VL domain and VH domain of an antibody combine to form the variable region (Fv) that defines a three dimensional antigen binding site. More specifically, the antigen binding site is defined by three complementary determining regions (CDRs) on each of the heavy and light chain variable regions. As used herein, the term “antigen binding site” includes a site that specifically binds an antigen (e.g., a cell surface or soluble antigen). The antigen binding site includes an immunoglobulin heavy chain and light chain variable region and the binding site formed by these variable regions determines the specificity of the antibody. An antigen binding site is formed by variable regions that vary from one antibody to another. The altered antibodies of the current disclosure comprise at least one antigen binding site.

[0093] In certain embodiments, binding polypeptides of the current disclosure comprise at least two antigen binding domains that provide for the association of the binding polypeptide with the selected antigen. The antigen binding domains need not be derived from the same immunoglobulin molecule. In this regard, the variable region may or be derived from any type of animal that can be induced to mount a humoral response and generate immunoglobulins against the desired antigen. As such, the variable region of a binding polypeptide may be, for example, of mammalian origin e.g., may be human, murine, rat, goat, sheep, non-human primate (such as cynomolgus monkeys, macaques, etc.), lupine, or camelid (e.g., from camels, llamas and related species).

[0094] In naturally occurring antibodies, the six CDRs present on each monomeric antibody are short, non-contiguous sequences of amino acids that are specifically positioned to form the antigen binding site as the antibody assumes its three-dimensional configuration in an aqueous environment. The remainder of the heavy and light variable domains show less inter-molecular variability in amino acid sequence and are termed the framework regions. The framework regions largely adopt a [3-sheet conformation and the CDRs form loops which connect, and in some cases form part of, the [3-sheet structure. Thus, these framework regions act to form a scaffold that provides for positioning the six CDRs in correct orientation by interchain, non-covalent interactions. The antigen binding domain formed by theAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP positioned CDRs defines a surface complementary to the epitope on the immunoreactive antigen. This complementary surface promotes the non-covalent binding of the antibody to the immunoreactive antigen epitope.

[0095] Exemplary binding polypeptides include antibody variants. As used herein, the term “antibody variant” includes synthetic and engineered forms of antibodies which are altered such that they are not naturally occurring, e.g., antibodies that comprise at least two heavy chain portions but not two complete heavy chains (such as, domain deleted antibodies or minibodies); multi-specific forms of antibodies (e.g., bi-specific, tri-specific, etc.) altered to bind to two or more different antigens or to different epitopes on a single antigen); heavy chain molecules joined to scFv molecules and the like. In addition, the term “antibody variant” includes multivalent forms of antibodies (e.g., trivalent, tetravalent, etc., antibodies that bind to three, four or more copies of the same antigen).

[0096] As used herein the term “valency” refers to the number of potential target binding sites in a polypeptide. Each target binding site specifically binds one target molecule or specific site on a target molecule. When a polypeptide comprises more than one target binding site, each target binding site may specifically bind the same or different molecules (e.g., may bind to different ligands or different antigens, or different epitopes on the same antigen). The subject binding polypeptides typically has at least one binding site specific for a human antigen molecule. For example, a typical lgG1 monoclonal antibody is specific for one target antigen. A bivalent antibody is one comprising antigen binding domains that targets two different antigens, or two antigen binding domains that target one antigen. Similarly, a trivalent antibody may be a monospecific antibody with three targeting domains to a single antigen. A trivalent antibody may be bispecific if it binds a first antigen with two binding domains and a second antigen with another binding domain. A trivalent antibody maybe trispecific and bind to three different targets.

[0097] The term “specificity” refers to the ability to specifically bind (e.g., immunoreact with) a given target antigen (e.g., a human target antigen). A binding polypeptide may be monospecific and contain one or more binding sites which specifically bind a target or a polypeptide may be multi-specific and contain two or more binding sites which specifically bind the same or different targets. In certainAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP embodiments, a binding polypeptide is specific for two different (e.g., nonoverlapping) portions of the same target. In certain embodiments, a binding polypeptide is specific for more than one target. Exemplary binding polypeptides (e.g., antibodies) which comprise antigen binding sites that bind to antigens expressed on tumor cells are known in the art and one or more CDRs from such antibodies can be included in an antibody as described herein.

[0098] The term “antigen” or “target antigen,” as used herein, refers to a molecule or a portion of a molecule that is capable of being bound by the binding site of a binding polypeptide. A target antigen may have one or more epitopes.

[0099] The term “about” or “approximately” means within about 20%, such as within about 10%, within about 5%, or within about 1% or less of a given value or range.

[0100] As used herein, “administer” or “administration” refers to the act of injecting or otherwise physically delivering a substance as it exists outside the body (e.g., an isolated binding polypeptide provided herein) into a patient, such as by, but not limited to, pulmonary (e.g., inhalation), mucosal (e.g., intranasal), intradermal, intravenous, intramuscular, subcutaneous delivery and / or any other method of physical delivery described herein or known in the art. When a disease, or a symptom thereof, is being managed or treated, administration of the substance typically occurs after the onset of the disease or symptoms thereof. When a disease, or symptom thereof, is being prevented, administration of the substance typically occurs before the onset of the disease or symptoms thereof and may be continued chronically to defer or reduce the appearance or magnitude of disease-associated symptoms.

[0101] As used herein, the term “composition” is intended to encompass a product containing the specified ingredients (e.g., an isolated binding polypeptide provided herein) in, optionally, the specified amounts, as well as any product which results, directly or indirectly, from combination of the specified ingredients in, optionally, the specified amounts.

[0102] “Effective amount” means the amount of active pharmaceutical agent (e.g., an isolated binding polypeptide of the present disclosure) sufficient to effectuate a desired physiological outcome in an individual in need of the agent. TheAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP effective amount may vary among individuals depending on the health and physical condition of the individual to be treated, the taxonomic group of the individuals to be treated, the formulation of the composition, assessment of the individual's medical condition, and other relevant factors.

[0103] As used herein, the terms “subject” and “patient” are used interchangeably. As used herein, a subject can be a mammal, such as a nonprimate (e.g., cows, pigs, horses, cats, dogs, rats, etc.) or a primate (e.g., monkey and human). In certain embodiments, the term “subject,” as used herein, refers to a vertebrate, such as a mammal. Mammals include, without limitation, humans, nonhuman primates, wild animals, feral animals, farm animals, sport animals, and pets.

[0104] As used herein, the term “therapy” refers to any protocol, method and / or agent that can be used in the prevention, management, treatment and / or amelioration of a disease or a symptom related thereto. In some embodiments, the term “therapy” refers to any protocol, method and / or agent that can be used in the modulation of an immune response to an infection in a subject or a symptom related thereto. In some embodiments, the terms “therapies” and “therapy” refer to a biological therapy, supportive therapy, and / or other therapies useful in the prevention, management, treatment and / or amelioration of a disease or a symptom related thereto, known to one of skill in the art such as medical personnel. In other embodiments, the terms “therapies” and “therapy” refer to a biological therapy, supportive therapy, and / or other therapies useful in the modulation of an immune response to an infection in a subject or a symptom related thereto known to one of skill in the art such as medical personnel.

[0105] As used herein, the terms “treat,” “treatment” and “treating” refer to the reduction or amelioration of the progression, severity, and / or duration of a disease or a symptom related thereto, resulting from the administration of one or more therapies (including, but not limited to, the administration of one or more prophylactic or therapeutic agents, such as an isolated binding polypeptide provided herein). The term “treating,” as used herein, can also refer to altering the disease course of the subject being treated. Therapeutic effects of treatment include, without limitation, preventing occurrence or recurrence of disease, alleviation of symptom(s), diminishment of direct or indirect pathological consequences of the disease,Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP decreasing the rate of disease progression, amelioration or palliation of the disease state, and remission or improved prognosis.

[0106] BCMA

[0107] As used herein, the term "BCMA" refers to B-cell maturation antigen. BCMA (also known as TNFRSF17, BCM or CD269) is a member of the tumor necrosis receptor (TNFR) family and is predominantly expressed on terminally differentiated B cells, e.g., memory B cells, and plasma cells. Its ligands are called B-cell activator of the TNF family (BAFF) and a proliferation inducing ligand (APRIL). BCMA is involved in mediating the survival of plasma cells for maintaining long-term humoral immunity. The gene for BCMA is encoded on chromosome 16 producing a primary 35 mRNA transcript of 994 nucleotides in length (NCBI accession NM_001192.2) that encodes a protein of 184 amino acids (NP_001183.2). A second antisense transcript derived from the BCMA locus has been described, which may play a role in regulating BCMA expression. (Laabi Y. et al. , Nucleic Acids Res. , 1994, 22:1147-1154.) Additional transcript variants have been described with unknown significance (Smirnova A S et al. Mol Immunol., 2008, 45(4): 1179-1183 A second isoform, also known as TV4, has been identified (Uniprot identifier Q02223-2). “BCMA” includes proteins comprising mutations, e.g., point mutations, fragments, insertions, deletions, and splice variants of full length wild-type BCMA.

[0108] Natural Killer Cells

[0109] As used herein, “natural killer cells” or “NK cells” refers to a subpopulation of lymphocytes that is involved in innate immunity. NK cells can be identified by virtue of certain characteristics and biological properties, such as the expression of specific surface antigens including CD16, CD56 and / or CD57, NKp46 for human NK cells, the absence of the alpha / beta or gamma / delta TCR complex on the cell surface, the ability to bind to and kill cells that fail to express “self’ MHC / HLA antigens by the activation of specific cytolytic machinery, the ability to kill tumor cells or other diseased cells that express a ligand for NK activating receptors, and the ability to release protein molecules called cytokines that stimulate or inhibit the immune response. Any of these characteristics and activities can be used toAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP identify NK cells, using methods well known in the art. Any subpopulation of NK cells will also be encompassed by the term NK cells. Within the context herein “active” NK cells designate biologically active NK cells, including NK cells having the capacity of lysing target cells or enhancing the immune function of other cells. NK cells can be obtained by various techniques known in the art, such as isolation from blood samples, cytapheresis, tissue or cell collections, etc. Useful protocols for assays involving NK cells can be found in Natural Killer Cells Protocols (edited by Campbell KS and Colonna M). Human Press, pp. 219-238 (2000).

[0110] NKp46

[0111] As used herein, the “NKp46” marker, or “natural cytotoxicity triggering receptor 1,” also known as “CD335” or “NKP46” or “NK-p46” or “LY94” refers to a protein or polypeptide encoded by the Ncr1 gene. A reference sequence of full-length human NKp46 protein is available from the NCBI database under the accession number NP_004820. The human NKp46 mRNA sequence is described in NCBI accession number NM_004829.

[0112] NK Cell Engager

[0113] As used herein, the isolated effector-competent polypeptide comprises a multispecific antibody in an NK cell engager format. An “NK cell engager” refers to binding proteins comprising monoclonal antibody domains targeting activating NK cell receptors, antigen-specific targeting regions, and optionally an Fc region (Gauthier, et al. (2019) Cell, 177: 1701-13). NK cells express CD16a, also known as FcyRllla, which binds with low affinity to the Fc parts of antibodies (Cerwenka and Lanier (2018) Science 359:6383). Engagement of CD16a is less demanding compared to CD3 engagement due to lower steric hindrances and is additionally facilitated by the lack of accessory molecules. Upon recognizing a target cell decorated with antibodies, NK cells mediate antibody-dependent cellular cytotoxicity (ADCC) resulting in killing of target cells (Lo Nigro (2019) Ann Transl Med 7:105). This naturally occurring mechanism can be utilized to engage CD16a when in conjunction with NKp46 (another activating NK cell receptor) to generate a trifunctional natural killer cell engager (i.e., NKCE) yielding impressive therapeuticAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP outcomes. For a review of NK cell engagers, see Demaria et al. (2021) Eur. J. Immunology 51(8): 1934, incorporated in its entirety by reference herein.

[0114] Trifunctional NKCEs are more potent in vitro than clinical therapeutic antibodies targeting the same antigen and they also have similar in vivo pharmacokinetics to full IgG antibodies and no off-target effects. See International Application No. PCT / IB21 / 62494 incorporated in its entirety by reference herein.

[0115] Fc Domains

[0116] In certain aspects of the disclosure, Fc domains, e.g., Fc domain variants, are provided. As used herein, the term “Fc region” or “Fc domain” refers to the portion of a heavy chain constant region beginning in the hinge region just upstream of the papain cleavage site (i.e., residue 216 in IgG, taking the first residue of heavy chain constant region to be 114) and ending at the C-terminus of the antibody. Accordingly, a complete Fc region comprises at least a hinge domain, a CH2 domain, and a CH3 domain.

[0117] The Fc region of an antibody is involved in non-antigen binding and can mediate effector function by binding to a Fc receptor. There are several different types of Fc receptors, which are classified based on the type of antibody that they recognize. For example, Fc-gamma receptors (FcyR) bind to IgG class antibodies, Fc-alpha receptors (FcaR) bind to IgA class antibodies, and Fc-epsilon receptors (FcsR) bind to IgE class antibodies. The neonatal Fc receptor (FcRn) interacts with the Fc region of an antibody to promote antibody recycling through rescue of normal lysosomal degradation. The FcyRs belong to a family that includes several members, e.g., FcyRI, FcyRlla, FcyRllb, FcyRllla, and FcyRlllb.

[0118] The term “native Fc” or “wild-type Fc,” as used herein, refers to a molecule corresponding to the sequence of a non-antigen-binding fragment resulting from digestion of an antibody or produced by other means, whether in monomeric or multimeric form, and can contain the hinge region. The original immunoglobulin source of the native Fc is typically of human origin and can be any of the immunoglobulins, such as lgG1 and lgG2. Native Fc molecules are made up of monomeric polypeptides that can be linked into dimeric or multimeric forms by covalent (i.e., disulfide bonds) and non-covalent association. The number ofAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP intermolecular disulfide bonds between monomeric subunits of native Fc molecules ranges from 1 to 4 depending on class (e.g., IgG, IgA, and IgE) or subclass (e.g., lgG1, lgG2, lgG3, lgA1, and lgGA2). One example of a native Fc is a disulfide-bonded dimer resulting from papain digestion of an IgG. The term “native Fc,” as used herein, is generic to the monomeric, dimeric, and multimeric forms.

[0119] The term “Fc domain variant,” “Fc variant” or “modified Fc,” as used herein, refers to a molecule or sequence that is modified from a native / wild-type Fc but still comprises a binding site for a FcR. Thus, the term “Fc variant” can comprise a molecule or sequence that is humanized from a non-human native Fc. Furthermore, a native Fc comprises regions that can be removed because they provide structural features or biological activities that are not required for the antibody-like binding polypeptides described herein. Thus, the term “Fc variant” comprises a molecule or sequence that lacks one or more native Fc sites or residues, or in which one or more Fc sites or residues has been modified, that affect or are involved in: (1) disulfide bond formation, (2) incompatibility with a selected host cell, (3) N-terminal heterogeneity upon expression in a selected host cell, (4) glycosylation, (5) interaction with complement, (6) binding to a Fc receptor other than a salvage receptor, or (7) antibody-dependent cellular cytotoxicity (ADCC).

[0120] As used herein, an “effector-competent Fc variant” or “effector-competent polypeptide” refers to a Fc domain that has one or more Fc effector functions as described further herein.

[0121] In certain exemplary embodiments, a Fc variant featured herein has one or more of increased serum half-life, enhanced FcRn binding affinity, enhanced FcRn binding affinity at acidic pH, enhanced FcyRllla binding affinity, and / or similar thermal stability, as compared to a wild-type Fc.

[0122] FcyRllla V176, or FcyRllla V158, or human CD16a-V receptor, or CD16av, refers to a polypeptide construct comprising a fragment of the CD16 human receptor binding to a Fc region of a natural antibody, mediating antibodydependent cellular cytotoxicity and bearing a Valine (V) on position 176 or position 158, which is also reported in the literature as allotype CD16a V176 or allotype CD16a V158.Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP FcyRllla F176, or FcyRllla F158, or human CD16a-F receptor, or CD16aF, refers to a polypeptide construct comprising a fragment of the CD16 human receptor binding to a Fc region of a natural antibody, mediating antibody-dependent cellular cytotoxicity and bearing a Phenylalanine (F) on position 176 or position 158, which is also reported in the literature as allotype CD16a F176 or allotype CD16a F158.

[0123] The term “Fc domain” as used herein encompasses native / wild-type Fc and Fc variants and sequences as defined herein. As with Fc variants and native Fc molecules, the term “Fc domain” includes molecules in monomeric or multimeric form, whether digested from whole antibody or produced by other means.

[0124] In certain exemplary embodiments, a Fc domain as described herein is thermally stabilized.

[0125] In certain exemplary embodiments, a Fc domain as described herein is glycosylated (e.g., via N--linked glycosylation). In certain exemplary embodiments, a Fc domain comprises N-linked glycosylation, e.g., at an N-linked glycosylation motif that contains the amino acid sequence NXT or NXS (X being any amino acid residue except proline). In certain exemplary embodiments, a Fc domain is glycosylated at amino acid position 297, according to Ell numbering.

[0126] In certain exemplary embodiments, a Fc domain as described herein is effector-competent.

[0127] In certain exemplary embodiments, a Fc domain as described herein is any combination of thermally stabilized, glycosylated, and effector-competent.

[0128] Thermally-Stabilized Fc Domain Variants

[0129] The structure of constant antibody domains is similar to that of the variable domains consisting of [3-strands connected with loops and short helices. The CH2 domain of the heavy constant regions exhibits weak carbohydrate-mediated interchain protein-protein interactions in contrast to the extensive interchain interactions exhibited in other domains. Isolated murine CH2 domains are relatively unstable at physiological temperature (Feige et al., 2004, J. Mol. Biol.

[0130] 344:107-118), but previous efforts demonstrate that the thermostability of CH2 domains may be enhanced with the addition of intrachain disulfide bonds, and thatAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP these could be used as scaffolds for binders (Gong et al., 2009, J. Biol. Chem.

[0131] 284:14203-210).

[0132] Effector-enhancing Fc domain variants that exhibit increased thermal instability (i.e., decreased thermal stability) relative to a wild-type Fc domain are known. For example, S239D / I332E and S239D / I332E / A330L variants lead to decreased stability of the CH2 domain as indicated by the lowering of melting temperature (Tm) in differential scanning calorimetry (DSC) analysis. G236A / S239D / A330L / I332E has a reduced protein thermal shift measurement when compared to wild-type, as well as a considerably reduced half-life in hFcyR transgenic mice. (See Liu et al. (2014) J. Biol. Chem. 289(6): 3571, and Liu et al. (2020) Antibodies 9(4): 64 for review.)

[0133] Effector-enhancing Fc domain variants having improved FcyR binding wherein stability is not significantly reduced as compared to wild-type are known. (See, e.g., Igawa et al., EP 2940 135)

[0134] It has been further discovered that thermostabilized Fc domain variants may be produced by introducing one or more disulfide bonds in the Fc domain. Accordingly, in one aspect, the present disclosure provides a Fc domain variant comprising one or more engineered (e.g., non-native) disulfide bonds, e.g., intrachain disulfide bonds mediated, e.g., by one or more pairs of cysteines.

[0135] In certain exemplary embodiments, a disulfide bond is an intrachain disulfide bond between the two CH2 regions of a Fc domain. In certain exemplary embodiments, a disulfide bond is an intrachain disulfide bond between the two CH3 regions of a Fc domain. In certain exemplary embodiments, two or more intrachain disulfide bonds are present in between the two CH2 regions of a Fc domain and / or between the two CH3 regions of a Fc domain.

[0136] Thermal stability, or the propensity of a Fc domain (e.g., a Fc domain with or without a binding polypeptide) to unfold, may be determined using a variety of methods known in the art. For example, the unfolding or denaturation temperature can be measured by nano-format differential scanning calorimetry (nanoDSC) or nano-format differential scanning fluorimetry (nanoDSF) (Wen et al., 2020 Anal. Biochem. 593:113581). The detectable temperature at which a protein begins to unfold is the Tonset.Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP In certain exemplary embodiments, the Tonset of a thermally-stabilized Fc domain variant (e.g., having one or more engineered disulfide bonds) is increased relative to a Fc domain variant that is not thermally stabilized. In certain exemplary embodiments, the Tonset of a thermally-stabilized Fc domain variant is increased by about 1.0 °C, about 1.5 °C, about 2.0 °C, about 2.5 °C, about 3.0 °C, about 3.5 °C, about 4.0 °C, about 4.5 °C, about 5.0 °C, about 5.5 °C, about 6.0 °C, about 6.5 °C, about 7.0 °C, about 7.5 °C, about 8.0 °C, about 8.5 °C, about 9.0 °C, about 9.5 °C, about 10.0 °C, about 10.5 °C, about 11.0 °C, about 11.5 °C, about 12.0 °C, about 12.5 °C, about 13.0 °C, about 13.5 °C, about 14.0 °C, about 14.5 °C, about 15.0 °C, about 15.5 °C, about 16.0 °C, about 16.5 °C, about 17.0 °C, about 17.5 °C, about 18.0 °C, about 18.5 °C, about 19.0 °C, about 19.5 °C, about 20.0 °C, about 20.5 °C, about 21.0 °C, about 21.5 °C, about 22.0 °C, about 22.5 °C, about 23.0 °C, about 23.5 °C, about 24.0 °C, about 24.5 °C or about 25.0 °C relative to a Fc domain variant that is not thermally stabilized.

[0137] In certain exemplary embodiments, a thermally-stabilized Fc domain variant has one or more amino acid substitution pairs selected from the group consisting of cysteine substitutions at: amino acid positions 242 and 334; amino acid positions 240 and 334; amino acid positions 287 and 306; amino acid positions 292 and 302; amino acid positions 323 and 332; amino acid positions 259 and 306; amino acid positions 350 and 441; amino acid positions 343 and 431; amino acid positions 375 and 404; amino acid positions 375 and 396; and amino acid positions 348 and 439, according to Ell numbering. (See Wozniak-Knopp et al., 2012, PLoS One 7: e30083; Jacobsen etal., 2017 J. Biol. Chem. 202:1865-75; WO2014153063 for reviews.)

[0138] In certain exemplary embodiments, a thermally-stabilized Fc domain variant comprises an engineered (e.g., a non-native) intrachain disulfide bond mediated by a pair of cysteines that substitute for (i) a leucine (L) at amino acid position 242 and a lysine (K) at amino acid position 334; (ii) an alanine (A) at amino acid position 287 and a leucine (L) at amino acid position 306; or (iii) an arginine (R) at amino acid position 292 and a valine (V) at amino acid position 302, according to Ell numbering.Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP In certain exemplary embodiments, a thermostabilized Fc domain variant comprises an engineered (e.g., a non-native) intrachain disulfide bond mediated by a pair of cysteines that substitute for (i) a leucine (L) at amino acid position 242 and a lysine (K) at amino acid position 334; (ii) an arginine (R) at amino acid position 292 and a valine (V) at amino acid position 302, according to Ell numbering.

[0139] In certain exemplary embodiments, a thermally-stabilized Fc domain variant comprises an engineered (e.g., a non-native) intrachain disulfide bond mediated by a pair of cysteines that substitute for a leucine (L) at amino acid position 242 and a lysine (K) at amino acid position 334. In certain exemplary embodiments, a thermally-stabilized Fc domain variant comprises an engineered (e.g., a non-native) intrachain disulfide bond mediated by a pair of cysteines that substitute an alanine (A) at amino acid position 287 and a leucine (L) at amino acid position 306. In certain exemplary embodiments, a thermally-stabilized Fc domain variant comprises an engineered (e.g., a non-native) intrachain disulfide bond mediated by a pair of cysteines that substitute for an arginine (R) at amino acid position 292 and a valine (V) at amino acid position 302. In certain exemplary embodiments, a thermally-stabilized Fc domain variant may comprise at least one engineered intrachain disulfide bond. In certain exemplary embodiments, a thermally-stabilized Fc domain variant may comprise more than one engineered intrachain disulfide bond.

[0140] Effector-Enhancing Fc Domain Variants

[0141] In one aspect, the present disclosure provides a Fc domain variant comprising effector-enhancing amino acid substitutions.

[0142] In one embodiment, a Fc domain variant with altered FcyRllla binding comprising one or more amino acid substitutions as disclosed herein. In one embodiment, a Fc domain variant with enhanced FcyRllla binding affinity having one or more amino acid substitutions as disclosed herein. In one embodiment, a Fc domain variant with enhanced FcyRllla binding affinity comprises two or more amino acid substitutions as disclosed herein. In one embodiment, a Fc domain variant with enhanced FcyRllla binding affinity comprises three or more amino acid substitutions as disclosed herein. In one embodiment, a Fc domain variant with enhancedAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP FcyRllla binding affinity comprises four or more amino acid substitutions as disclosed herein.

[0143] In one embodiment, a Fc domain variant with altered FcRn binding comprises a Fc domain having one or more amino acid substitutions as disclosed herein. In one embodiment, a Fc domain variant with enhanced FcRn binding affinity comprises a Fc domain having one or more amino acid substitutions as disclosed herein. In one embodiment, a Fc domain variant with enhanced FcRn binding affinity comprises a Fc domain having two or more amino acid substitutions as disclosed herein. In one embodiment, a Fc domain variant with enhanced FcRn binding affinity comprises a Fc domain having three or more amino acid substitutions as disclosed herein.

[0144] In some embodiments, a Fc domain variant may exhibit a speciesspecific FcRn binding affinity. In one embodiment, a Fc domain variant may exhibit hum a FcRn binding affinity. In one embodiment, a Fc domain variant may exhibit cyno FcRn binding affinity. In some embodiments, a Fc domain variant may exhibit cross-species FcRn binding affinity. Such a Fc domain variants are said to be cross-reactive across one or more different species. In one embodiment, a Fc domain variant may exhibit both human and cynomolgus monkey (cyno) FcRn binding affinity.

[0145] The neonatal Fc receptor (FcRn) interacts with the Fc region of antibodies to promote recycling through rescue of normal lysosomal degradation. This process is a pH-dependent process that occurs in the endosomes at acidic pH (e.g., a pH less than 6.5) but not under the physiological pH conditions of the bloodstream (e.g., a non-acidic pH). In some embodiments, a Fc domain variant has enhanced FcRn binding affinity at an acidic pH compared to a wild-type Fc domain. In some embodiments, a Fc domain variant has enhanced FcRn binding affinity at pH less than 7.0, e.g., at about pH 6.5, at about pH 6.0, at about pH 5.5, at about pH 5.0, compared to a wild-type Fc domain. In some embodiments, a Fc domain variant has enhanced FcRn binding affinity at pH less than 7.0, e.g., at about pH 6.5, at about pH 6.0, at about pH 5.5, at about pH 5.0, compared to the FcRn binding affinity of a wild-type Fc domain at an elevated non-acidic pH. An elevated non-Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP acidic pH can be, e.g., pH greater than 7.0, about pH 7.0, about pH 7.4, about pH 7.6, about pH 7.8, about pH 8.0, about pH 8.5, about pH 9.0.

[0146] In certain embodiments, it may be desired for a Fc domain variant to exhibit approximately the same FcRn binding affinity at non-acidic pH as a wild-type Fc domain. In some embodiments, it may be desired for a Fc domain variant to exhibit less FcRn binding affinity at non-acidic pH than a binding polypeptide comprising a modified Fc domain having the double amino acid substitution M428L / N434S, according to Ell numbering (See LISPN 8,088,376). Accordingly, it may be desired a Fc domain variant to exhibit minimal perturbation to pH-dependent FcRn binding.

[0147] In some embodiments, a Fc domain variant having enhanced FcRn binding affinity at an acidic pH, has a reduced (i.e., slower) FcRn off-rate as compared to a wild-type Fc domain. In some embodiments, a Fc domain variant having enhanced FcRn binding affinity at an acidic pH compared to the FcRn binding affinity of the binding polypeptide at an elevated non-acidic pH, has a slower FcRn off-rate at the acidic pH compared to the FcRn off-rate of a wild-type Fc domain at the elevated non-acidic pH.

[0148] Certain embodiments include Fc domain variants in which at least one amino acid in one or more of the constant region domains has been deleted or otherwise altered so as to provide desired biochemical characteristics such as reduced or enhanced effector functions, the ability to non-covalently dimerize, increased ability to localize at the site of a tumor, reduced serum half-life, or increased serum half-life when compared with a whole, unaltered antibody of approximately the same immunogenicity.

[0149] In certain other embodiments, a Fc domain variant comprises constant regions derived from different antibody isotypes (e.g., constant regions from two or more of a human lgG1, lgG2, lgG3, or lgG4). In other embodiments, a Fc domain variant comprises a chimeric hinge (i.e., a hinge comprising hinge portions derived from hinge domains of different antibody isotypes, e.g., an upper hinge domain from an lgG4 molecule and an lgG1 middle hinge domain). In certain embodiments, the Fc domain may be mutated to increase or decrease effector function using techniques known in the art.Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP In some embodiments, a Fc domain variant has altered binding affinity to a Fc receptor. There are several different types of Fc receptors, which are classified based on the type of antibody that they recognize. For example, Fc-gamma receptors (FcyR) bind to IgG class antibodies, Fc-alpha receptors (FcaR) bind to IgA class antibodies, and Fc-epsilon receptors (FcsR) bind to IgE class antibodies. The FcyRs belong to a family that includes several members, e.g., FcyRI, FcyRlla, FcyRllb, FcyRllla, and FcyRlllb. In some embodiments, a Fc domain variant has altered FcyRllla binding affinity, compared to a wild-type Fc domain. In some embodiments, a Fc domain variant has reduced FcyRllla binding affinity, compared to a wild-type Fc domain. In some embodiments, a Fc domain variant has enhanced FcyRllla binding affinity, compared to a wild-type Fc domain. In some embodiments, a Fc domain variant modified Fc domain has approximately the same FcyRllla binding affinity, compared to a wild-type Fc domain.

[0150] In certain embodiments, a Fc domain variant comprises an antibody constant region (e.g., an IgG constant region e.g., a human IgG constant region, e.g., a human lgG1 constant region) which mediates one or more effector functions. For example, binding of the C1 -complex to an antibody constant region may activate the complement system. Activation of the complement system is important in the opsonization and lysis of cell pathogens. The activation of the complement system also stimulates the inflammatory response and may also be involved in autoimmune hypersensitivity. Furthermore, antibodies bind to receptors on various cells via the Fc domain (Fc receptor binding sites on the antibody Fc region bind to Fc receptors (FcRs) on a cell). There are a number of Fc receptors which are specific for different classes of antibody, including IgG (gamma receptors), IgE (epsilon receptors), IgA (alpha receptors) and IgM (mu receptors). Binding of antibody to Fc receptors on cell surfaces triggers a number of important and diverse biological responses including engulfment and destruction of antibody-coated particles, clearance of immune complexes, lysis of antibody-coated target cells by killer cells (called antibody-dependent cell-mediated cytotoxicity, or ADCC), release of inflammatory mediators, placental transfer and control of immunoglobulin production. In some embodiments, a Fc domain variant, e.g., a binding polypeptide (e.g., an antibody, immunoadhesin or an antibody variant) binds to a Fc-gamma receptor. In alternativeAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP embodiments, a Fc domain variant comprised a constant region which is devoid of one or more effector functions (e.g., ADCC activity) and / or is unable to bind Fey receptor.

[0151] In certain exemplary embodiments, an effector-enhancing Fc domain variant has one or more amino acid substitutions selected from the group consisting of: an aspartic acid (D) at amino acid position 221; a cysteine (C) at amino acid position 222; a tyrosine (Y) at amino acid position 234; an alanine (A) at amino acid position 236; a tryptophan (W) at amino acid position 236; an aspartic acid (D) at amino acid position 239; a leucine (L) at amino acid position 243; a glutamic acid (E) at amino acid position 267; a phenylalanine (F) at amino acid position 268; a proline (P) at amino acid position 292; an alanine (A) at amino acid position 298; a leucine (L) at amino acid position 300; an isoleucine (I) at amino acid position 305; a threonine (T) at amino acid position 324; a tryptophan (W) at amino acid position 326; an alanine (A) at amino acid position 326; a leucine (L) at amino acid position 330; a glutamic acid (E) at amino acid position 332; an alanine (A) at amino acid position 333; a serine (S) at amino acid position 333; an alanine (A) at amino acid position 334; an alanine (A) at amino acid position 336; an arginine (R) at amino acid position 345; and a leucine (L) at amino acid position 396, according to Ell numbering. (See Saunders, 2009, Front. Immunol, doi: 10.3389 / fimmu.2019.01296, for a review.)

[0152] In some embodiments, a Fc domain variant may comprise an amino acid substitution at positions selected from amino acid positions 236, 239, 330, and 332, according to Ell numbering. In some embodiments, the substitutions may comprise an alanine (A) at amino acid position 236, an aspartic acid (D) at amino acid 239, a leucine (L) at amino acid position 330, and a glutamic acid (E) at amino acid position 332, according to Ell numbering. In some embodiments, a Fc domain variant may comprise a double amino acid substitution at any two amino acid positions selected from an alanine (A) at amino acid position 236, aspartic acid (D) at amino acid 239, a leucine (L) at amino acid position 330, and a glutamic acid (E) at amino acid position 332. In some embodiments, a Fc domain variant may comprise a triple amino acid substitution at any three amino acid positions selected from an alanine (A) at amino acid position 236, an aspartic acid (D) at amino acid 239, a leucine (L)Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP at amino acid position 330, and a glutamic acid (E) at amino acid position 332. In some embodiments, a Fc domain variant may comprise a quadruple amino acid substitution at any four amino acid positions selected from an alanine (A) at amino acid position 236, an aspartic acid (D) at amino acid 239, a leucine (L) at amino acid position 330, and a glutamic acid (E) at amino acid position 332. In some embodiments, a Fc domain variant may comprise the combination of amino acid substitutions comprising an aspartic acid (D) at amino acid 239 and a glutamic acid (E) at amino acid position 332. In some embodiments, a Fc domain variant may comprise the combination of amino acid substitutions comprising an alanine (A) at amino acid position 236, an aspartic acid (D) at amino acid position and a glutamic acid at position 332.

[0153] In some embodiments, a Fc domain variant may further comprise an amino acid substitution at amino acid positions 256 and / or 307, according to Ell numbering. In some embodiments, a Fc domain variant may comprise the combination of amino acid substitutions comprising an aspartic acid (D) at amino acid positions 256 and a glutamine (Q) at amino acid position 307 (See Mackness et al., 2019 MAbs 11:1276-88; WO2019147973A1 , incorporated in its entirety by reference herein).

[0154] Glycosylated Fc Domain Variants

[0155] In certain exemplary embodiments, a binding protein is glycosylated. Glycosylation of antibodies at conserved positions in their constant regions is known to have a profound effect on antibody function, particularly effector functioning such as those described above, see for example, Boyd et al (Mol. Immunol, 32: 1311-1318, 1996). Glycosylation of a binding protein of the present disclosure wherein one or more carbohydrate moiety is added, substituted, deleted or modified are contemplated. In some embodiments, the glycosylation of the Fc domain of the binding protein is an N-linked glycosylation. Introduction of an asparagine-X-serine or asparagine-X-threonine motif creates a potential site for enzymatic attachment of carbohydrate moieties and may therefore be used to manipulate the glycosylation of a Fc domain variant. In Raju et al. (Biochemistry 40: 8868-8876, 2001) the terminal sialylation of a TNFR-IgG immunoadhesin was increased through aAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP process of re-galactosylation and / or re-sialylation using (3-1 ,4-galactosyltransferace and / or alpha, 2,3 sialyltransferase. Increasing the terminal sialylation is believed to increase the half-life of the immunoglobulin.

[0156] Antibodies, in common with most glycoproteins, are typically produced as a mixture of glycoforms. This mixture is particularly apparent when antibodies are produced in eukaryotic, particularly mammalian cells. A variety of methods have been developed to manufacture defined glycoforms (see Zhang et al. 2004, Science 303: 371; Sears et al, 2001, Science 291: 2344; Wacker et al., 2002, Science 298: 1790; Davis et al. 2002, Chem. Rev. 102: 579; Hang et al., 2001, Acc. Chem. Res.

[0157] 34: 727). In some embodiments, the glycosylated Fc domain comprises a native glycan at amino acid position 297, according to Ell numbering. Glycosylation of the asparagine at amino acid position 297 in the CH2 domain of lgG1 is known to facilitate interaction between the Fc domain and FcyR. Elimination of this glycosylation site eliminates effector function (Leabman, et al., 2013, MAbs 5:896-903). In particularly exemplary embodiments, a Fc domain comprises wild-type levels, or near wild-type levels, of glycosylation at amino acid position 297, according to Ell numbering.

[0158] In some embodiments, the glycosylated Fc domain variant comprises an engineered or non-native glycan. In some embodiments, the engineered or nonnative glycan is a modified glycan that can be conjugated to a therapeutic molecule (e.g., antibody-drug conjugate).

[0159] Fc-Containing Binding Polypeptides

[0160] In one aspect, the present disclosure provides an isolated Fc domain variant comprising or complexed with (e.g., fused to) at least one binding domain (e.g., at least one binding polypeptide). In certain embodiments, the binding domain comprises one or more antigen binding domains. The antigen binding domains need not be derived from the same molecule as the parental Fc domain. In certain embodiments, the Fc domain variant is present in an antibody.

[0161] In one embodiment, a Fc domain variant is present in an antibody or is complexed with an antibody. Any antibody from any source or species can be employed with a Fc domain variant disclosed herein. Suitable antibodies includeAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP without limitation, chimeric antibodies, humanized antibodies, or human antibodies. Suitable antibodies include without limitation, full-length antibodies, monoclonal antibodies, polyclonal antibodies, or single-domain antibodies, such as VHH antibodies.

[0162] In certain exemplary embodiments, a Fc domain variant may be bound to or complexed with an antigen-binding fragment of an antibody. The term “antigenbinding fragment” refers to a polypeptide fragment of an immunoglobulin or antibody which binds antigen or competes with intact antibody (i.e., with the intact antibody from which they were derived) for antigen binding (i.e., specific binding). Antigenbinding fragments can be produced by recombinant or biochemical methods that are well known in the art. Exemplary antigen-binding fragments include Fv, Fab, Fab', and (Fab')2. In certain exemplary embodiments, a binding polypeptide of the current disclosure comprises at least one antigen-binding fragment and a Fc domain variant.

[0163] In some embodiments, the binding polypeptide comprises a single chain variable region sequence (ScFv). Single chain variable region sequences comprise a single polypeptide having one or more antigen binding sites, e.g., a VL domain linked by a flexible linker to a VH domain. ScFv molecules can be constructed in a VH-linker-VL orientation or VL-linker-VH orientation. The flexible hinge that links the VL and VH domains that make up the antigen binding site includes from about 10 to about 50 amino acid residues. Connecting peptides are known in the art. Binding polypeptides may comprise at least one scFv and / or at least one constant region. In one embodiment, a binding polypeptide of the current disclosure may comprise at least one scFv linked or fused to a Fc domain variant.

[0164] In some embodiments, a binding polypeptide of the current disclosure is a multivalent (e.g., tetravalent) antibody which is produced by fusing a DNA sequence encoding an antibody with a ScFv molecule (e.g., an altered ScFv molecule). For example, in one embodiment, these sequences are combined such that the ScFv molecule (e.g., an altered ScFv molecule) is linked at its N-terminus or C-terminus to a Fc domain variant via a flexible linker (e.g., a gly / ser linker). In another embodiment a tetravalent antibody of the current disclosure can be madeAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP by fusing a ScFv molecule to a connecting peptide, which is fused to a Fc domain variant to construct a ScFv-Fab tetravalent molecule.

[0165] In another embodiment, a binding polypeptide of the current disclosure is an altered minibody. An altered minibody of the current disclosure is a dimeric molecule made up of two polypeptide chains each comprising a ScFv molecule which is fused to a Fc domain variant via a connecting peptide. Minibodies can be made by constructing a ScFv component and connecting peptide components using methods described in the art (see, e.g., US patent 5,837,821 or WO 94 / 09817AI). In another embodiment, a tetravalent minibody can be constructed. Tetravalent minibodies can be constructed in the same manner as minibodies, except that two ScFv molecules are linked using a flexible linker. The linked scFv-scFv construct is then joined to a Fc domain variant.

[0166] In another embodiment, a binding polypeptide of the current disclosure comprises a diabody. Diabodies are dimeric, tetravalent molecules each having a polypeptide similar to scFv molecules, but usually having a short (less than 10, e.g., about 1 to about 5) amino acid residue linker connecting both variable domains, such that the VL and VH domains on the same polypeptide chain cannot interact. Instead, the VL and VH domain of one polypeptide chain interact with the VH and VL domain (respectively) on a second polypeptide chain (see, for example, WO 02 / 02781 ). Diabodies of the current disclosure comprise a scFv-like molecule fused to a Fc domain variant.

[0167] In another embodiment, a binding polypeptide of the current disclosure comprises a single-domain antibody (sdAb), also referred to as a VHH or a nanobody. Nanobody® is registered trademark of Ablynx. VHHs comprise variable heavy chain domains devoid of light chains. Similar to conventional VH domains, VHHs contain four FRs and three CDRs. VHHs have advantages over conventional antibodies. As they are about ten times smaller than IgG molecules, properly folded functional VHHs can be produced by in vitro expression while achieving high yield. Furthermore, VHHs are very stable, and resistant to the action of proteases. The properties and production of VHHs have been reviewed by Harmsen and De Haard H J (Appl. Microbiol. Biotechnol. 2007 November; 77(1): 13-22).Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP In certain exemplary embodiments, a Fc domain is fused with one or more VHHs.

[0168] In other embodiments, binding polypeptides comprise multi-specific or multivalent antibodies comprising one or more variable domain in series on the same polypeptide chain, e.g., tandem variable domain (TVD) polypeptides. Exemplary TVD polypeptides include the “double head” or “dual-Fv” configuration described in U.S. Patent No. 5,989,830. In the dual-Fv configuration, the variable domains of two different antibodies are expressed in a tandem orientation on two separate chains (one heavy chain and one light chain), wherein one polypeptide chain has two VH domains in series optionally separated by a peptide linker (VH1-linker-VH2) and the other polypeptide chain consists of complementary VL domains optionally connected in series by a peptide linker (VL1-linker-VL2). In the cross-over double head configuration, the variable domains of two different antibodies are expressed in a tandem orientation on two separate polypeptide chains (one heavy chain and one light chain), wherein one polypeptide chain has two VH domains in series optionally separated by a peptide linker (VH1-linker-VH2) and the other polypeptide chain consists of complementary VL domains optionally connected in series by a peptide linker in the opposite orientation (VL2-linker-VL1). Additional antibody variants based on the “dual-Fv” format include the dual-variable-domain IgG (DVD-IgG) bispecific antibody (see U.S. Patent No. 7,612,181 and the TBTI format (see US 2010 / 0226923 A1). In some embodiments, binding polypeptides comprise multi-specific or multivalent antibodies comprising one or more variable domain in series on the same polypeptide chain fused to a Fc domain variant.

[0169] In another embodiment, a binding polypeptide comprises a cross-over dual variable domain IgG (CODV-IgG) bispecific antibody based on a “double head” configuration (see US20120251541 A1, which is incorporated by reference herein in its entirety).

[0170] In other embodiments, a binding polypeptide comprises a CrossMab or a CrossMab-Fab multispecific format (see W02009080253 and Schaefer, et al., PNAS (2011 ), 108: 11187-1191 ). Antibody variants based on the CrossMab format have a crossover of antibody domains within one arm of a bispecific IgG antibody enabling correct chain association.Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP In other embodiments, the glycosylated effector-competent polypeptide comprises a multispecific antibody in a T cell engager format. A “T cell engager” refers to binding proteins directed to a host’s immune system, more specifically the T cells’ cytotoxic activity as well as directed to a tumor target protein. In some embodiments, the isolated effector-competent polypeptide comprises a multispecific antibody in a NK cell engager format. A “NK cell engager” refers to binding proteins comprising monoclonal antibody fragments targeting activating NK cell receptors, antigen-specific targeting regions, and a Fc region (Gauthier, etal. Cell (2019), 177: 1701-13).

[0171] A binding polypeptide of the present disclosure, comprising a Fc domain variant described herein, can include the CDR sequences or the variable domain sequences of a known “parent” antibody. In some embodiments, the parent antibody and the antibody of the disclosure can share similar or identical sequences except for modifications to the Fc domain as disclosed herein.

[0172] Cross-Over Dual Variable

[0173] In a particular embodiment, “cross-over dual variable” or “CODV” refers to an antigen-binding domain that specifically binds to at least one target antigen or at least one target epitope, and comprises at least two polypeptide chains that form at least two antigen-binding sites, wherein at least one polypeptide chain comprises a structure represented by the formula:

[0174] VL1-L1-VL2-L2-CL [I]

[0175] and at least one polypeptide chain comprises a structure represented by the formula:

[0176] VH2-L3-VH1-L4-CH1 [II]

[0177] wherein:

[0178] VL1 is a first immunoglobulin light chain variable domain;

[0179] VL2 is a second immunoglobulin light chain variable domain;

[0180] VH1 is a first immunoglobulin heavy chain variable domain;

[0181] VH2 is a second immunoglobulin heavy chain variable domain;

[0182] CL is an immunoglobulin light chain constant domain;

[0183] CH1 is an immunoglobulin CH1 heavy chain constant domain; andAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP L1 , L2, L3, and L4 are amino acid linkers, wherein any one or more of L1 , L2, L3, and L4 are optionally absent, and

[0184] wherein the polypeptides of formula I and the polypeptides of formula II form a cross-over light chain-heavy chain pair.

[0185] In certain exemplary embodiments, the binding protein of the disclosure comprises a “CODV-OL1” format, comprising three polypeptide chains that form two antigen-binding sites, wherein one polypeptide chain comprises a structure represented by the formula:

[0186] VL1-L1-VL2-L2-CL [I];

[0187] one polypeptide chain comprises a structure represented by the formula: VH2-L3-VH1-L4-CH1-hinge-CH2-CH3 [III]; and

[0188] one polypeptide chain comprises a structure represented by the formula: hinge-CH2-CH3 [IV]

[0189] wherein:

[0190] CL is an immunoglobulin light chain constant domain;

[0191] 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;

[0192] and

[0193] L1 , L2, L3, and L4 are amino acid linkers, wherein any one or more of L1 , L2, L3, and L4 are optionally absent, and

[0194] wherein the polypeptides of formula I and the polypeptides of formula II form a cross-over light chain-heavy chain pair.

[0195] In a particular embodiment, a CODV antigen-binding domain specifically binds to at least one target antigen or at least one target epitope, and comprises four polypeptide chains that form four antigen-binding sites, wherein two polypeptide chains each comprises a structure represented by the formula:

[0196] VL1-L1-VL2-L2-CL [I]

[0197] and two polypeptide chains each comprises a structure represented by the formula:Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP VH2-L3-VH1-L4-CH1-Fc [II]

[0198] wherein:

[0199] VL1 is a first immunoglobulin light chain variable domain;

[0200] VL2 is a second immunoglobulin light chain variable domain;

[0201] VH1 is a first immunoglobulin heavy chain variable domain;

[0202] VH2 is a second immunoglobulin heavy chain variable domain;

[0203] CL is an immunoglobulin light chain constant domain;

[0204] CH1 is an immunoglobulin CH1 heavy chain constant domain;

[0205] Fc is an immunoglobulin hinge region and CH2 and CH3 immunoglobulin heavy chain constant domains; and

[0206] L1 , L2, L3, and L4 are amino acid linkers, wherein any one or more of L1 , L2, L3, and L4 are optionally absent, and

[0207] wherein the polypeptides of formula I and the polypeptides of formula II form a cross-over light chain-heavy chain pair,

[0208] wherein the VH1 / VL1 pair comprises a first antigen binding specificity and the VH2 / VL2 pair comprises a second antigen binding specificity.

[0209] In a particular embodiment, an antigen-binding protein described herein is a trispecific and / or a trivalent antigen-binding protein comprising four polypeptide chains that form three antigen-binding sites that specifically bind to one or more different antigen targets, wherein the first polypeptide chain comprises a structure represented by the formula:

[0210] VL2-L1-VL1-L2-CL [I]

[0211] the second polypeptide chain comprises a structure represented by the formula:

[0212] VH1 -L3-VH2-L4-CH1 -hinge-CH2-CH3 [II]

[0213] the third polypeptide chain comprises a structure represented by the formula:

[0214] VH3-CH1-hinge-CH2-CH3 [III]

[0215] and the fourth polypeptide chain comprises a structure represented by the formula:

[0216] VL3-CL [IV],

[0217] wherein:Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP VL1 is a first immunoglobulin light chain variable domain;

[0218] VL2 is a second immunoglobulin light chain variable domain;

[0219] VL3 is a third immunoglobulin light chain variable domain;

[0220] VH1 is a first immunoglobulin heavy chain variable domain;

[0221] VH2 is a second immunoglobulin heavy chain variable domain;

[0222] VH3 is a third immunoglobulin heavy chain variable domain;

[0223] CL is an immunoglobulin light chain constant domain;

[0224] CH1 is an immunoglobulin CH1 heavy chain constant domain;

[0225] CH2 is an immunoglobulin CH2 heavy chain constant domain;

[0226] CH3 is an immunoglobulin CH3 heavy chain constant domain; hinge is an immunoglobulin hinge region connecting the CH1 and CH2 domains; and

[0227] L1 , L2, L3, and L4 are amino acid linkers, wherein any one or more of L1 , L2, L3, and L4 are optionally absent, and

[0228] wherein the polypeptides of formula I and the polypeptides of formula II form a cross-over light chain-heavy chain pair.

[0229] In certain embodiments, the first polypeptide chain and the second polypeptide chain have a cross-over orientation that forms two distinct antigenbinding sites. In some embodiments, the VH1 and VL1 form a binding pair and form the first antigen-binding site. In some embodiments, the VH2 and VL2 form a binding pair and form the second antigen-binding site. In some embodiments, the third polypeptide and the fourth polypeptide form a third antigen-binding site. In some embodiments, the VH3 and VL3 form a binding pair and form the third antigenbinding site.

[0230] Such antigen-binding protein comprises at least three antigen-binding sites. It is at least a trivalent antigen-binding molecule. In a particular embodiment, it specifically binds to one antigen target, i.e., it is a monospecific antigen-binding molecule. In another embodiment, it specifically binds to two different antigen targets, i.e., it is a bispecific antigen-binding molecule. In another embodiment, it specifically binds to three different antigen targets, i.e., it is a trispecific antigenbinding molecule.Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP The examples listed above are not intended to limit the scope of the disclosure in any way, and linkers comprising randomly selected amino acids selected from the group consisting of valine, leucine, isoleucine, serine, threonine, lysine, arginine, histidine, aspartate, glutamate, asparagine, glutamine, glycine, and proline have been shown to be suitable in the antibody-like binding proteins described herein.

[0231] In certain embodiments of the binding protein, (a) L1 , L2, L3, and L4 each independently are zero amino acids in length or comprise a sequence selected from the group consisting of GGGGSGGGGS, GGGGSGGGGSGGGGS, S, RT, TKGPS, GQPKAAP, and GGSGSSGSGG; or (b) L1, L2, L3, and L4 each independently comprise a sequence selected from the group consisting of GGGGSGGGGS, GGGGSGGGGSGGGGS, S, RT, TKGPS, GQPKAAP, and GGSGSSGSGG.

[0232] In certain embodiments, L1 and L2 each comprise the amino acid sequence GGGGSGGGGS.

[0233] In certain embodiments, wherein L3 and L4 are each absent.

[0234] The CODV antibody format, the various permutations of the CODV antibody format, and additional details regarding linkers is further described in WO 2012 / 135345A1 , and WO 2017 / 180913A2, which are incorporated herein by reference in their entireties.

[0235] BCMA NKCE for the Treatment of B-Cell Mediated Autoimmune Diseases Autoimmune diseases occur when the immune system mistakenly attacks the body's own tissues and organs, leading to inflammation and damage. This can involve various immune cells, including autoreactive B cells and plasma cells, which produce antibodies that target self-antigens (e.g., autoantibodies). Examples of B-cell or plasma cell mediated autoimmune diseases include rheumatoid arthritis (RA) and lupus, such as systemic lupus erythematosus (SLE).

[0236] The term “rheumatoid arthritis” refers to a chronic, autoimmune disease that primarily affects the joints, although other tissues can be affected. Although rheumatoid arthritis is mediated by a variety of immune cells, evidence has shown an involvement of B cells and autoantibodies (see, Wu et al. Front Immunol. 2021.

[0237] 12: 750753).Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP The term “rheumatoid factor” or “RF” refers to an autoantibody that is commonly elevated in rheumatoid arthritis patients. RF binds the Fc portion of IgG, forming an immune complex that may contribute to rheumatoid arthritis progression.

[0238] The term “lupus” is a chronic autoimmune disease that occurs when the body's immune system attacks its own tissues and organs. The term “SLE” is a chronic autoimmune disease with multisystemic involvement. SLE is the most common type of lupus. SLE manifestations are associated with multiple autoantibodies against DNA or other nuclear components forming immune complexes that deposit in various organs. SLE can affect many parts of the body, such as the skin, joints, tissues, kidneys, brain, and other organs. Symptoms of SLE can range from mild to life-threatening. It was found that BCMA expression was significantly increased in patients with autoimmune diseases, such as SLE (Vincent et al. Clin. Transl. Immunol. 8 e1047 (2019)), which correlates with disease activity (Laurent et al. Nat. Commun. 6:7333 (2015)).

[0239] Immunosuppressive treatments such as corticosteroids, diseasemodifying antirheumatic drugs (DMARDs), and biologic therapies can be effective in controlling symptoms, reducing inflammation, and preventing disease progression in autoimmune diseases. However, not all patients respond to these treatments, and some may experience significant side effects. Thus, there exists a need in the art for potent therapies in treating B-cell mediated autoimmune diseases.

[0240] The disclosure provides methods of treating an autoimmune disorder (e.g., a B-cell mediated autoimmune disorder) in a subject with a binding protein that binds a Natural Killer (NK) cell marker and BCMA described herein.

[0241] Thus, in one aspect, the disclosure provides a method of treating a B-cell mediated autoimmune disorder in a subject, comprising administering to the subject a binding protein comprising a first antigen binding domain (ABD) with binding specificity to BCMA and a second ABD with binding specificity to NKp46, thereby treating the B-cell mediated autoimmune disorder in the subject.

[0242] In certain embodiments, the methods of the invention include treatment of patients having lupus. In certain embodiments, the lupus is SLE. In certain embodiments, the subject has a reduced serum immunoglobulin level compared to a subject that was not administered a binding protein comprising a first ABD withAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP binding specificity to BCMA and a second ABD with binding specificity to NKp46. In certain embodiments, the subject has reduced IgA, IgM, and / or IgG serum immunoglobulin levels compared to a subject that was not administered a binding protein comprising a first ABD with binding specificity to BCMA and a second ABD with binding specificity to NKp46.

[0243] In certain embodiments, the methods of the invention include treatment of patients having rheumatoid arthritis (RA). In certain embodiments, the subject has a reduced serum immunoglobulin level compared to a subject that was not administered a binding protein comprising a first ABD with binding specificity to BCMA and a second ABD with binding specificity to NKp46. In certain embodiments, the subject has reduced IgA, IgM, and / or IgG serum immunoglobulin levels compared to a subject that was not administered a binding protein comprising a first ABD with binding specificity to BCMA and a second ABD with binding specificity to NKp46. In certain embodiments, the subject has a reduced serum rheumatoid factor (RF) level compared to a subject that was not administered a binding protein comprising a first ABD with binding specificity to BCMA and a second ABD with binding specificity to NKp46.

[0244] Pharmaceutical Compositions and Administration Thereof

[0245] Methods of preparing and administering the binding protein of the current disclosure are well known to or are readily determined by those skilled in the art. The route of administration of the binding polypeptides of the current disclosure may be oral, parenteral, by inhalation or topical. The term parenteral as used herein includes intravenous, intraarterial, intraperitoneal, intramuscular, subcutaneous, rectal or vaginal administration. While all these forms of administration are clearly contemplated as being within the scope of the current disclosure, a form for administration would be a solution for injection, in particular for intravenous or intraarterial injection or drip or for subcutaneous administration. Usually, a suitable pharmaceutical composition for injection may comprise a buffer (e.g., acetate, phosphate or citrate buffer), a surfactant (e.g., polysorbate), optionally a stabilizer agent (e.g., human albumin), etc. In some embodiments, Fc domain variants can beAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP delivered directly to the site of the adverse cellular population thereby increasing the exposure of the diseased tissue to the therapeutic agent.

[0246] Preparations for parenteral administration include sterile aqueous or nonaqueous solutions, suspensions, and emulsions. Aqueous carriers include water, alcoholic / aqueous solutions, emulsions or suspensions, including saline and buffered media. In the compositions and methods of the current disclosure, pharmaceutically acceptable carriers include, but are not limited to, 0.01-0.1 M, e.g., 0.05 M phosphate buffer, or 0.8% saline. Other common parenteral vehicles include sodium phosphate solutions, Ringer’s dextrose, dextrose and sodium chloride, lactated Ringer’s, or fixed oils. Intravenous vehicles include fluid and nutrient replenishers, electrolyte replenishers, such as those based on Ringer’s dextrose, and the like. Preservatives and other additives may also be present such as for example, antimicrobials, antioxidants, chelating agents, and inert gases and the like. More particularly, pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In such cases, the composition must be sterile and should be fluid to the extent that easy syringability exists. It should be stable under the conditions of manufacture and storage and will typically be preserved against the contaminating action of microorganisms, such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.

[0247] In many cases, isotonic agents will be included, for example, sugars, polyalcohols, such as mannitol, sorbitol, or sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate and gelatin.

[0248] In any case, sterile injectable solutions can be prepared by incorporating an active compound (e.g., a binding protein of the disclosure) in the required amountAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP in an appropriate solvent with one or a combination of ingredients enumerated herein, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle, which contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, exemplary methods of preparation include vacuum drying and freeze-drying, which yields a powder of an active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof. The preparations for injections are processed, filled into containers such as ampoules, bags, bottles, syringes or vials, and sealed under aseptic conditions according to methods known in the art. Further, the preparations may be packaged and sold in the form of a kit. Such articles of manufacture will typically have labels or package inserts indicating that the associated compositions are useful for treating a subject suffering from or predisposed to autoimmune or neoplastic disorders.

[0249] Effective doses of the compositions of the present disclosure, for the treatment of the above-described conditions vary depending upon many different factors, including means of administration, target site, physiological state of the patient, whether the patient is human or an animal, other medications administered, and whether treatment is prophylactic or therapeutic. Usually, the patient is a human, but non-human mammals including transgenic mammals can also be treated. Treatment dosages may be titrated using routine methods known to those of skill in the art to optimize safety and efficacy.

[0250] A pharmaceutical composition in accordance with the present disclosure can comprise a pharmaceutically acceptable, non-toxic, sterile carrier such as physiological saline, nontoxic buffers, preservatives and the like. For the purposes of the instant application, a pharmaceutically effective amount of the binding protein shall be held to mean an amount sufficient to achieve effective binding to an antigen and to achieve a benefit, e.g., to ameliorate symptoms of a disease or disorder or to detect a substance or a cell. In the case of tumor cells, the polypeptide can interact with selected antigens on neoplastic or immunoreactive cells and provide for an increase in the death of those cells. Of course, the pharmaceutical compositions of the present disclosure may be administered in single or multipleAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP doses to provide for a pharmaceutically effective amount of the modified binding polypeptide.

[0251] In keeping with the scope of the present disclosure, the binding proteins of the disclosure may be administered to a human or other animal in accordance with the aforementioned methods of treatment in an amount sufficient to produce a therapeutic or prophylactic effect. The binding proteins of the disclosure can be administered to such human or other animal in a conventional dosage form prepared by combining the antibody of the disclosure with a conventional pharmaceutically acceptable carrier or diluent according to known techniques. It will be recognized by one of skill in the art that the form and character of the pharmaceutically acceptable carrier or diluent is dictated by the amount of active ingredient with which it is to be combined, the route of administration and other well-known variables. Those skilled in the art will further appreciate that a cocktail comprising one or more species of binding polypeptides described in the current disclosure may prove to be particularly effective.

[0252] The contents of the articles, patents, and patent applications, and all other documents and electronically available information mentioned or cited herein, are hereby incorporated by reference in their entirety to the same extent as if each individual publication was specifically and individually indicated to be incorporated by reference. Applicants reserve the right to physically incorporate into this application any and all materials and information from any such articles, patents, patent applications, or other physical and electronic documents.

[0253] While the present disclosure has been described with reference to the specific embodiments thereof, it should be understood by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the true spirit and scope of the disclosure. It will be readily apparent to those skilled in the art that other suitable modifications and adaptations of the methods described herein may be made using suitable equivalents without departing from the scope of the embodiments disclosed herein. In addition, many modifications may be made to adapt a particular situation, material, composition of matter, process, process step or steps, to the objective, spirit and scope of the present disclosure. All such modifications are intended to be within the scope of theAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP claims appended hereto. Having now described certain embodiments in detail, the same will be more clearly understood by reference to the following examples, which are included for purposes of illustration only and are not intended to be limiting.

[0254] EMBODIMENTS OF THE DISCLOSURE

[0255] 1. A method of treating an autoimmune disorder in a subject, comprising administering to the subject a binding protein comprising a first antigen binding domain (ABD) with binding specificity to BCMA and a second ABD with binding specificity to NKp46, thereby treating the B-cell mediated autoimmune disorder in the subject.

[0256] 2. The method of embodiment 1 , wherein:

[0257] (a) the first ABD comprises:

[0258] (a1) a first immunoglobulin heavy chain variable domain (VH1) comprising an HCDR1 comprising the amino acid sequence GFTFSNFGMH (SEQ ID NO: 1), an HCDR2 comprising the amino acid sequence VIWSDETNR (SEQ ID NO: 2), and an HCDR3 comprising the amino acid sequence DQQYCSSDSCFTWFDP (SEQ ID NO: 3); and

[0259] (a2) a first immunoglobulin light chain variable domain (VL1) comprising an LCDR1 comprising the amino acid sequence CASSTGTVTPSNYAN (SEQ ID NO: 7), an LCDR2 comprising the amino acid sequence DNNSRPP (SEQ ID NO: 8), and an LCDR3 comprising the amino acid sequence ALWFGNQWV (SEQ ID NO: 9); or

[0260] (a) the first ABD comprises:

[0261] (a1) a first immunoglobulin heavy chain variable domain (VH1) comprising an HCDR1 comprising the amino acid sequence GFTFSNFG (SEQ ID NO: 112), an HCDR2 comprising the amino acid sequence IWSDETNR (SEQ ID NO: 113), and an HCDR3 comprising the amino acid sequence ARDQQYCSSDSCFTWFDP (SEQ ID NO: 114); andAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP (a2) a first immunoglobulin light chain variable domain (VL1) comprising an LCDR1 comprising the amino acid sequence TGTVTPSNY (SEQ ID NO: 118), an LCDR2 comprising the amino acid sequence DNN (SEQ ID NO: 119), and an LCDR3 comprising the amino acid sequence ALWFGNQWV (SEQ ID NO: 120); or

[0262] (a) the first ABD comprises:

[0263] (a1) a first immunoglobulin heavy chain variable domain (VH1) comprising an HCDR1 comprising the amino acid sequence NFGMH (SEQ ID NO: 115), an HCDR2 comprising the amino acid sequence VIWSDETNRYYADSVKG (SEQ ID NO: 116), and an HCDR3 comprising the amino acid sequence DQQYCSSDSCFTWFDP (SEQ ID NO: 117); and

[0264] (a2) a first immunoglobulin light chain variable domain (VL1) comprising an LCDR1 comprising the amino acid sequence ASSTGTVTPSNYAN (SEQ ID NO: 121), an LCDR2 comprising the amino acid sequence DNNSRPP (SEQ ID NO: 122), and an LCDR3 comprising the amino acid sequence ALWFGNQWV (SEQ ID NO: 123).

[0265] 3. The method of embodiment 1 or 2, wherein (b) the second ABD comprises:

[0266] (b1) a second immunoglobulin heavy chain variable domain (VH2) comprising:

[0267] - an HCDR1 comprising the amino acid sequence DYVIN, an HCDR2 comprising the amino acid sequence EIYPGSGTNYYNEKFKA, and an HCDR3 comprising the amino acid sequence RGRYGLYAMDY;

[0268] - an HCDR1 sequence comprising GYTFSDYVIN (SEQ ID NO: 19), an HCDR2 sequence comprising EIYPGSGTN (SEQ ID NO: 20), and an HCDR3 sequence comprising RGRYGLYAMDY (SEQ ID NO: 21);

[0269] - an HCDR1 comprising the amino acid sequence SDYAWN (SEQ ID NO: 22), an HCDR2 comprising the amino acid sequence YITYSGSTSYNPSLES (SEQ ID NO: 23), and an HCDR3 comprising the amino acid sequence GGYYGSSWGVFAY (SEQ ID NO: 24);Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP - an HCDR1 comprising the amino acid sequence EYTMH (SEQ ID NO: 25), an HCDR2 comprising the amino acid sequence GISPNIGGTSYNQKFKG (SEQ ID NO: 26), and an HCDR3 comprising the amino acid sequence RGGSFDY (SEQ ID NO: 27);

[0270] - an HCDR1 comprising the amino acid sequence SFTMH (SEQ ID NO: 28), an HCDR2 comprising the amino acid sequence YINPSSGYTEYNQKFKD (SEQ ID NO: 29), and an HCDR3 comprising the amino acid sequence GSSRGFDY (SEQ ID NO: 30); or

[0271] - an HCDR1 comprising the amino acid sequence SDYAWN (SEQ ID NO: 31), an HCDR2 comprising the amino acid sequence YITYSGSTNYNPSLKS (SEQ ID NO: 32), and an HCDR3 comprising the amino acid sequence CWDYALYAMDC (SEQ ID NO: 33); and

[0272] (b2) a second immunoglobulin light chain variable domain (VL2) comprising:

[0273] - an LCDR1 comprising the amino acid sequence RASQDISNYLN (SEQ ID NO: 34), an LCDR2 comprising the amino acid sequence YTSRLHS (SEQ ID NO: 35), and an LCDR3 comprising the amino acid sequence QQGNTRPWT (SEQ ID NO: 36);

[0274] - an LCDR1 comprising the amino acid sequence RVSENIYSYLA (SEQ ID NO: 37), an LCDR2 comprising the amino acid sequence NAKTLAE (SEQ ID NO: 38), and an LCDR3 comprising the amino acid sequence QHHYGTPWT (SEQ ID NO: 39);

[0275] - an LCDR1 comprising the amino acid sequence RASQSISDYLH (SEQ ID NO: 40), an LCDR2 comprising the amino acid sequence YASQSIS (SEQ ID NO: 41), and an LCDR3 comprising the amino acid sequence QNGHSFPLT (SEQ ID NO: 42);

[0276] - an LCDR1 comprising the amino acid sequence RASENIYSNLA (SEQ ID NO: 43), an LCDR2 comprising the amino acid sequence AATNLAD (SEQ ID NO: 44), and an LCDR3 comprising the amino acid sequence QHFWGTPRT (SEQ ID NO: 45); orAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP - an LCDR1 comprising the amino acid sequence RTSENIYSYLA (SEQ ID NO: 46), an LCDR2 comprising the amino acid sequence NAKTLAE (SEQ ID NO: 47), and an LCDR3 comprising the amino acid sequence QHHYDTPLT (SEQ ID NO: 48).

[0277] 4. The method of embodiment 2 or 3, wherein the VL1 comprises:

[0278] - an LCDR1 sequence comprising the amino acid sequence of CASSTGTVTPSNYAN (SEQ ID NO: 7), an LCDR2 sequence comprising the amino acid sequence of DNNSRPP (SEQ ID NO: 8), and an LCDR3 sequence comprising the amino acid sequence of ALWFGNQWV (SEQ ID NO: 9);

[0279] - an LCDR1 sequence comprising the amino acid sequence of CRSSTGTVTPSNYAN (SEQ ID NO: 10), an LCDR2 sequence comprising the amino acid sequence of DNNSRPP (SEQ ID NO: 11), and an LCDR3 sequence comprising the amino acid sequence of ALWFGNQWV (SEQ ID NO: 12);

[0280] - an LCDR1 sequence comprising the amino acid sequence of CASSTGAVTPSNYAN (SEQ ID NO: 13), an LCDR2 sequence comprising the amino acid sequence of DNNIKPP (SEQ ID NO: 14), and an LCDR3 sequence comprising the amino acid sequence of ALWYGGQWV (SEQ ID NO: 15); or

[0281] - an LCDR1 sequence comprising the amino acid sequence of CASSTGAVTPGYYAN (SEQ ID NO: 16), an LCDR2 sequence comprising the amino acid sequence of DNNNKPP (SEQ ID NO: 17), and an LCDR3 sequence comprising the amino acid sequence of ALYYGGQWV (SEQ ID NO: 18).

[0282] 5. The method of embodiment 1 , wherein:

[0283] (a) the first ABD comprises:

[0284] (a1) a first immunoglobulin heavy chain variable domain (VH1) comprising an HCDR1 sequence comprising the amino acid sequence of GFTFSNFG (SEQ ID NO: 112), an HCDR2 sequence comprising the amino acid sequence of IWSDETNR (SEQ ID NO: 113), and an HCDR3 sequence comprising the amino acid sequence of ARDQQYCSSDSCFTWFDP (SEQ ID NO: 114); and (a2) a first immunoglobulin light chain variable domain (VL1) comprising an LCDR1 sequence comprising the amino acid sequence of TGTVTPSNY (SEQAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP ID NO: 118), an LCDR2 sequence comprising the amino acid sequence of DNN (SEQ ID NO: 119), and an LCDR3 sequence comprising the amino acid sequence of ALWFGNQWV (SEQ ID NO: 120).

[0285] 6. The method of embodiment 1 , wherein:

[0286] (a) the first ABD comprises:

[0287] (a1) a first immunoglobulin heavy chain variable domain (VH1) comprising an HCDR1 sequence comprising the amino acid sequence of NFGMH (SEQ ID NO: 115), an HCDR2 sequence comprising the amino acid sequence of VIWSDETNRYYADSVKG (SEQ ID NO: 116), and an HCDR3 sequence comprising the amino acid sequence of DQQYCSSDSCFTWFDP (SEQ ID NO: 117); and (a2) a first immunoglobulin light chain variable domain (VL1) comprising an LCDR1 sequence comprising the amino acid sequence of ASSTGTVTPSNYAN (SEQ ID NO: 121), an LCDR2 sequence comprising the amino acid sequence of DNNSRPP (SEQ ID NO: 122), and an LCDR3 sequence comprising the amino acid sequence of ALWFGNQWV (SEQ ID NO: 123).

[0288] 7. The method of any one of embodiments 2-6, wherein:

[0289] - the VH1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 55;

[0290] - the VH1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 50;

[0291] - the VH1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 51 ;

[0292] - the VH1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 49, and wherein the VL1Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 52;

[0293] - the VH1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 53; or

[0294] - the VH1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 54.

[0295] 8. The method of any one of embodiments 2-7, wherein:

[0296] - the VH1 comprises an amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence of SEQ ID NO: 55;

[0297] - the VH1 comprises an amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence of SEQ ID NO: 50;

[0298] - the VH1 comprises an amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence of SEQ ID NO: 51 ;

[0299] - the VH1 comprises an amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence of SEQ ID NO: 52;

[0300] - the VH1 comprises an amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence of SEQ ID NO: 53; or

[0301] - the VH1 comprises an amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence of SEQ ID NO: 54.

[0302] 9. The method of any one of embodiments 3-8, wherein:

[0303] - the VH2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 56, and wherein the VL2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 64;

[0304] - the VH2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 57, and wherein the VL2Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 65;

[0305] - the VH2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 58, and wherein the VL2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 66;

[0306] - the VH2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 59, and wherein the VL2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 67;

[0307] - the VH2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 60, and wherein the VL2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 68;

[0308] - the VH2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 61, and wherein the VL2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 69;

[0309] - the VH2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 62, and wherein the VL2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 70; or

[0310] - the VH2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 63, and wherein the VL2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 71.

[0311] 10. The method of any one of embodiments 3-9, wherein:

[0312] - the VH2 comprises an amino acid sequence of SEQ ID NO: 56, and wherein the VL2 comprises an amino acid sequence of SEQ ID NO: 64;

[0313] - the VH2 comprises an amino acid sequence of SEQ ID NO: 57, and wherein the VL2 comprises an amino acid sequence of SEQ ID NO: 65;Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP - the VH2 comprises an amino acid sequence of SEQ ID NO: 58, and wherein the VL2 comprises an amino acid sequence of SEQ ID NO: 66;

[0314] - the VH2 comprises an amino acid sequence of SEQ ID NO: 59, and wherein the VL2 comprises an amino acid sequence of SEQ ID NO: 67;

[0315] - the VH2 comprises an amino acid sequence of SEQ ID NO: 60, and wherein the VL2 comprises an amino acid sequence of SEQ ID NO: 68;

[0316] - the VH2 comprises an amino acid sequence of SEQ ID NO: 61, and wherein the VL2 comprises an amino acid sequence of SEQ ID NO: 69;

[0317] - the VH2 comprises an amino acid sequence of SEQ ID NO: 62, and wherein the VL2 comprises an amino acid sequence of SEQ ID NO: 70; or

[0318] - the VH2 comprises an amino acid sequence of SEQ ID NO: 63, and wherein the VL2 comprises an amino acid sequence of SEQ ID NO: 71.

[0319] 11. The method of any one of embodiments 1-10, wherein the binding protein further comprises all or part of an immunoglobulin Fc domain or variant thereof.

[0320] 12. The method of embodiment 11, wherein all or part of the immunoglobulin Fc domain or variant thereof binds to a human Fc-y receptor.

[0321] 13. The method of embodiment 11 or 12, wherein all or part of the immunoglobulin Fc domain or variant thereof binds to a human CD16A (FcyRIII) polypeptide.

[0322] 14. The method of any one of embodiments 11-13, wherein the Fc domain comprises a native glycan at amino acid position 297, according to Ell numbering.

[0323] 15. The method of any one of embodiments 11-14, wherein the binding protein is N-glycosylated.Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP 16. The method of any one of embodiments 11-15, wherein the Fc domain or variant thereof comprises a first Fc heavy chain and a second Fc heavy chain.

[0324] 17. The method of embodiment 16, wherein at least one Fc heavy chain comprises an engineered intrachain disulfide bond mediated by a pair of cysteines (C) that substitute for:

[0325] (i) a leucine (L) at amino acid position 242 and a lysine (K) at amino acid position 334;

[0326] or

[0327] (ii) an arginine (R) at amino acid position 292 and a valine (V) at amino acid position 302;

[0328] wherein the amino acid positions are according to Ell numbering.

[0329] 18. The method of embodiment 17, wherein the first and the second Fc heavy chain each comprise both the L242C and K334C substitutions.

[0330] 19. The method of embodiment 17, wherein the first and the second Fc heavy chain each comprise both the R292C and V302C substitutions.

[0331] 20. The method of any one of embodiments 16-19, wherein at least one Fc heavy chain comprises a substitution at amino acid position 332, according to Ell numbering.

[0332] 21. The method of embodiment 20, wherein the substitution at amino acid position 332 is a glutamic acid (E).

[0333] 22. The method of embodiment 20 or 21, wherein the at least one Fc heavy chain further comprises one or more substitutions at amino acid positions 236, 239, or 330, according to Ell numbering.Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP 23. The method of embodiment 22, wherein the substitution at amino acid position 236 is an alanine (A), the substitution at amino acid position 239 is an aspartic acid (D), and the substitution at amino acid position 330 is a leucine (L).

[0334] 24. The method of any one of embodiments 16-23, wherein at least one Fc heavy chain comprises an aspartic acid (D) at amino acid position 239, and a glutamic acid (E) at amino acid position 332, according to Ell numbering.

[0335] 25. The method of any one of embodiments 16-24, wherein at least one Fc heavy chain comprises an alanine (A) at amino acid position 236, an aspartic acid (D) at amino acid position 239, and a glutamic acid (E) at amino acid position 332, according to Ell numbering.

[0336] 26. The method of any one of embodiments 16-25, wherein at least one Fc heavy chain further comprises an alanine (A) at amino acid position 236, an aspartic acid (D) at amino acid position 239, a leucine (L) at amino acid position 330, and a glutamic acid (E) at amino acid position 332, according to Ell numbering.

[0337] 27. The method of any one of embodiments 3-26, wherein the binding protein comprising at least two polypeptide chains that form at least two antigenbinding sites, wherein at least one polypeptide chain comprises a structure represented by the formula:

[0338] VL1-L1-VL2-L2-CL [I];

[0339] and at least one polypeptide chain comprises a structure represented by the formula:

[0340] VH2-L3-VH1-L4-CH1 [II];

[0341] wherein:

[0342] CL is an immunoglobulin light chain constant domain;

[0343] CH1 is an immunoglobulin CH1 heavy chain constant domain; andAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP L1 , L2, L3, and L4 are amino acid linkers, wherein any one or more of L1 , L2, L3, and L4 are optionally absent, and

[0344] wherein the polypeptides of formula I and the polypeptides of formula II form a cross-over light chain-heavy chain pair.

[0345] 28. The method of any one of embodiments 3-27, wherein the binding protein comprises three polypeptide chains that form two antigen-binding sites, wherein one polypeptide chain comprises a structure represented by the formula:

[0346] VL1-L1-VL2-L2-CL [I];

[0347] one polypeptide chain comprises a structure represented by the formula: VH2-L3-VH1-L4-CH1-hinge-CH2-CH3 [III]; and

[0348] one polypeptide chain comprises a structure represented by the formula: hinge-CH2-CH3 [IV]

[0349] wherein:

[0350] CL is an immunoglobulin light chain constant domain;

[0351] 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;

[0352] and

[0353] L1 , L2, L3, and L4 are amino acid linkers, wherein any one or more of L1 , L2, L3, and L4 are optionally absent, and

[0354] wherein the polypeptides of formula I and the polypeptides of formula II form a cross-over light chain-heavy chain pair.

[0355] 29. The method of embodiment 27 or 28, wherein:

[0356] (a) L1 , L2, L3, and L4 each independently are zero amino acids in length or comprise a sequence selected from the group consisting of GGGGSGGGGS, GGGGSGGGGSGGGGS, S, RT, TKGPS, GQPKAAP, and GGSGSSGSGG; orAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP (b) L1 , L2, L3, and L4 each independently comprise a sequence selected from the group consisting of GGGGSGGGGS, GGGGSGGGGSGGGGS, S, RT, TKGPS, GQPKAAP, and GGSGSSGSGG.

[0357] 30. The method of any one of embodiments 1-29, wherein:

[0358] (a) the first ABD of the binding protein comprises a first immunoglobulin heavy chain variable domain (VH1) comprising an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 49, and a first immunoglobulin light chain variable domain (VL1) comprising an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 55; and

[0359] (b) the second ABD of the binding protein comprises a second immunoglobulin heavy chain variable domain (VH2) comprising an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 56, and a second immunoglobulin light chain variable domain (VL2) comprising an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 64.

[0360] 31. The method of any one of embodiments 1-30, wherein the binding protein comprises:

[0361] (i) a first polypeptide chain comprising an amino acid sequence of SEQ ID NO: 72;

[0362] (ii) a second polypeptide chain comprising an amino acid sequence of SEQ ID NO: 73; and

[0363] (iii) a third polypeptide chain comprising an amino acid sequence of SEQ ID NO: 74.

[0364] 32. The method of any one of embodiments 1 or 2, wherein:

[0365] (b) the second ABD comprises:

[0366] (b1) a second immunoglobulin heavy chain variable domain (VH2) comprising an HCDR1 sequence comprising the amino acid sequence of GYTFSDYV (SEQ ID NO: 87), an HCDR2 sequence comprising the amino acidAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP sequence of IYPGSGTN (SEQ ID NO: 90), and an HCDR3 sequence comprising the amino acid sequence of ARRGRYGLYAMDY (SEQ ID NO: 93); and

[0367] (b2) a second immunoglobulin light chain variable domain (VL1) comprising an LCDR1 sequence comprising the amino acid sequence of QDISNY (SEQ ID NO: 96), an LCDR2 sequence comprising the amino acid sequence of YTS (SEQ ID NO: 99), and an LCDR3 sequence comprising the amino acid sequence of QQGNTRPWT (SEQ ID NO: 111 ).

[0368] 33. The method of any one of embodiments 1 or 2, wherein:

[0369] (b) the second ABD comprises:

[0370] (b1) a second immunoglobulin heavy chain variable domain (VH2) comprising an HCDR1 sequence comprising the amino acid sequence of DYVIN (SEQ ID NO: 80), an HCDR2 sequence comprising the amino acid sequence of EIYPGSGTNYYNEKFKA (SEQ ID NO: 81), and an HCDR3 sequence comprising the amino acid sequence of RGRYGLYAMDY (SEQ ID NO: 21); and

[0371] (b2) a second immunoglobulin light chain variable domain (VL1) comprising an LCDR1 sequence comprising the amino acid sequence of RASQDISNYLN (SEQ ID NO: 34), an LCDR2 sequence comprising the amino acid sequence of YTSRLHS (SEQ ID NO: 35), and an LCDR3 sequence comprising the amino acid sequence of QQGNTRPWT (SEQ ID NO: 36).

[0372] 34. The method of any one of embodiments 1-33, wherein the autoimmune disorder is a B-cell mediated autoimmune disorder.

[0373] 35. The method of any one of embodiments 1-33, wherein the autoimmune disorder is a plasma cell mediated autoimmune disorder.

[0374] 36. The method of any one of embodiments 1-35, wherein the autoimmune disorder is lupus.

[0375] 37. The method of embodiment 36, wherein the lupus is systemic lupus erythematosus (SLE).Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0376] 38. The method of any one of embodiments 1-34, wherein the autoimmune disorder is rheumatoid arthritis (RA).

[0377] 39. The method of any one of embodiments 1-34, wherein the subject has a reduced serum immunoglobulin level compared to a subject that was not administered the binding protein.

[0378] 40. The method of any one of embodiments 1-34, wherein subject has a reduced autoantibody level compared to a subject that was not administered the binding protein.

[0379] 41. The method of embodiment 39 or 40, wherein the serum immunoglobulin and / or auntoantibody is IgA, IgM, and / or IgG.

[0380] 42. The method of any one of embodiments 1-41, wherein the subject has a reduced serum rheumatoid factor (RF) level compared to a subject that was not administered the binding protein.

[0381] 43. The method of any one of embodiments 1-42, wherein the binding protein is administered intravenously or subcutaneously.Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP EXAMPLES

[0382] The present disclosure is further illustrated by the following examples, which should not be construed as further limiting.

[0383] Example 1: Evaluation of Total Immunoglobulin (IgA, IgM and IgA) Concentrations in Serum of Cynomolgus Monkeys After Weekly Administration of BCMA NKCE (SAR445514) by SC and IV routes

[0384] Introduction:

[0385] The aim of this study is to determine total immunoglobulin (IgA, IgG, and IgM) concentrations in serum of cynomolgus monkeys following once weekly subcutaneous or intravenous (1-hour infusion) administration of NKP46-BCMA_FC-ADE-DSB NKCE (which is also known as BCMA NKCE or SAR445514) for 5 weeks.

[0386] Sequence for BCMA NKCE (SAR445514):

[0387] BCMA NKCE (SAR445514) is schematically depicted in FIG. 1 and comprises: a first antigen binding domain (ABD) with binding specificity to BCMA and a second ABD with binding specificity to NKp46, wherein:

[0388] (a) the first ABD of the binding protein comprises a first immunoglobulin heavy chain variable domain (VH1) comprising an amino acid sequence of SEQ ID NO: 49, and a first immunoglobulin light chain variable domain (VL1) comprising an amino acid sequence of SEQ ID NO: 55; and

[0389] (b) the second ABD of the binding protein comprises a second immunoglobulin heavy chain variable domain (VH2) comprising an amino acid sequence of SEQ ID NO: 56, and a second immunoglobulin light chain variable domain (VL2) comprising an amino acid sequence of SEQ ID NO: 64.

[0390] BCMA NKCE (SAR445514) has three polypeptide chains, (i) the first polypeptide chain comprising an amino acid sequence of SEQ ID NO: 72; (ii) the second polypeptide chain comprising an amino acid sequence of SEQ ID NO: 73; and (iii) the third polypeptide chain comprising an amino acid sequence of SEQ ID NO: 74.Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP Study Design:

[0391] Cynomolgus monkeys (3 monkeys per sex per group) received the control subcutaneously (bolus, 1 mL / kg) and intravenously (1-hour infusion, 5 mL / kg), or a solution of BCMA NKCE (SAR445514) at doses of 5, 15, or 49.1 mg / kg administered subcutaneously (SC) (1 mL / kg), or a solution of BCMA NKCE (SAR445514) at a dose of 30 mg / kg administered intravenously (IV) (5 mL / kg) on Days 1, 8, 15, 22, and 29 for a total of 5 weekly administrations. The study design is described in Table 1. Monkeys were scheduled to be euthanized and necropsied one week after the last (5th) administration on Day 36.

[0392] Table 1- Experimental Design

[0393]

[0394]

[0395] Total serum immunoglobulins evaluation

[0396] Serum samples were collected by venepuncture before each dosing (on Days 1, 8, 15, 22 and 29), 24 hours after the first dose (i.e. , Day 2), and 168 hoursAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP after the last dose (i.e., Day 36), centrifuged at 1800 g, during 10 min at approximately +4°C, and retained at -80°C until analysis. Samples analyses were performed using a commercially multiplex assay as described in Table 2.

[0397] Table 2- Total Immunoglobulin (IgA, IgG, and IgM) Determination in Serum and Kit components

[0398]

[0399] Kit components

[0400]

[0401] Immunoglobulins assays in the Isotyping Panel 1 (Human / NHP) kit from Meso Scale Discovery (MSD) are sandwich immunoassays based on the captureAttorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP on a plate precoated with specific anti-lgA, anti-IgG and anti-IgM antibodies on independent and well-defined spots of the microplate and based on the detection with a solution containing detection antibody conjugated with electrochemiluminescent MSD SULFO-TAG™ labels throughout several incubations. Immunoglobulins in the monkey serum sample

[0402] bind to capture antibodies immobilized on the working electrode surface and recruitment of the detection antibodies by the bound analytes completes the sandwich. Following MSD buffer that provides appropriate environment for electrochemiluminescence and voltage applied to the plate electrodes when loading plate in the MESO QuickPlex SQ 120, the capture labels emit light. The intensity of emitted light is proportional to the concentration of analytes in the sample.

[0403] The calibration range was defined from 781.25 to 200000.00 pg / mL for IgA, IgG and IgM. Study serum samples were 250000-fold diluted in the sample diluent of the kit (Diluent 100 containing PBS, blockers and preservatives) prior to the assay. The lower limit of quantification (LLOQ) for study serum samples was 0.20 mg / mL (781.25 pg / mL x dilution factor, 250 000). The upper limit of quantification (IILOQ) for study serum samples was 50.00 mg / mL (200 000.00 pg / mL x dilution factor, 250000).

[0404] There were no reagents provided in the kit to prepare control samples. Consequently, Quality Controls (QCs) of two different levels (QC Low and QC High) were prepared at a concentration corresponding approximately to a low and high concentration of the calibration curve prepared from the stock solution provided in the kit for the preparation of calibration samples diluted in the sample diluent as described in Table 3.

[0405] Table 3- Control Samples

[0406]

[0407] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0408] Each calibration standard sample, control sample and study sample were assayed in duplicate in one determination. Samples with coefficient of variation (CV (%)) superior to 20% were rejected. However, samples inferior to LLOQ were not submitted to such CV% criteria.

[0409] A calibration curve was considered as acceptable when a minimum of 7 individual standards including at least one replicate for the LLOQ and ULOQ corresponding to 70% of the 10 individual standards were accepted (% of recovery from the nominal value between 70 and 130% including LLOQ, ULOQ and CV (%) <20%).

[0410] For control samples, a run was considered as acceptable when at least 75% of QCs tested (QC High and Low, each in duplicate, one set at the start and one set at the end of the run) were within a given concentration level between 70% and 130% of recovery from the nominal value.

[0411] Results:

[0412] Study Sample Analysis

[0413] Individual serum concentrations (initially measured in pg / mL) are expressed in mg / mL and reported to 2 decimal places and expressed as percent-changes from baseline (i.e. predose or Day 1 -predose). Individual serum concentrations obtained below the LLOQ (0.20 mg / mL) were arbitrarily replaced by half of the LLOQ (i.e.

[0414] 0.10 mg / mL). Descriptive statistics including mean, standard deviation (SD) were calculated at each analyzed timepoint for each Ig, and for each dose group.

[0415] Interpretation was performed based on % changes from baseline. Changes in Ig concentrations were considered biologically significant when % changes from baseline were more than those observed in the control group throughout the study reaching globally ±30% of the baseline value (i.e. within the analytical variability range and biological variations).Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP IQA Concentrations

[0416] Baseline IgA concentrations observed on Day 1 , predose in both males and females reported in Table 4, ranged from 0.41 mg / mL to 2.22 mg / mL with a mean and SD values of 1.09 ±0.49 mg / mL.

[0417] Table 4- Baseline IgA, IgG, and IgM concentrations measured on Day 1 predose

[0418]

[0419] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0420]

[0421] i) BCMA NKCE (SAR445514) / Control Group

[0422] Individual serum IgA concentrations expressed in mg / mL and as percent-changes from baseline are reported in Table 5 for males and in Table 6 for females, respectively. Individual IgA concentrations across the study (from Day 1 , predose to Day 36) ranged from 0.75 to 1.76 mg / mL with a mean and SD values of 1.18 ±0.34 mg / mL for males and from 0.34 to 1.95 mg / mL with a mean and SD values of 0.72 ±0.45 mg / mL for females, respectively.

[0423] Table 5- Control Group Males: Serum IgA levels on Study Days

[0424]

[0425] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0426]

[0427] Table 6- Control Group Females: Serum IgA levels on Study Days

[0428]

[0429] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0430]

[0431] ii) BCMA NKCE (SAR445514) / Dosage 5 mg / kg / adm SC

[0432] Individual serum IgA concentrations expressed in mg / mL and as percent-changes from baseline are reported in Table 7 for males and in Table 8 for females, respectively. Individual IgA concentrations across the study (from Day 1 , predose to Day 36) ranged from 0.10 to 1.88 mg / mL with a mean and SD values of 0.79 ±0.63 mg / mL for males and from 0.10 to 1.70 mg / mL with a mean and SD values of 0.86 ±0.46 mg / mL for females, respectively. All animals showed reductions in IgA concentrations up to -88% (Female No. 10 on Day 36) occurring from Day 8 and Day 22 with a greater magnitude of change observed later in the dosing period at Day 36, except for one isolated male (Number 9) where IgA concentrations were maintained during the study within the biological variation range.

[0433] Table 7- BCMA NKCE (SAR445514) Dosage 5 mg / kg / adm SC- Males: Serum IgA Levels on Study Days

[0434]

[0435] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0436]

[0437] Table 8- BCMA NKCE (SAR445514) Dosage 5 mg / kg / adm SC- Females: Serum IgA Levels on Study Days

[0438]

[0439] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0440]

[0441] Hi) BCMA NKCE (SAR445514) / Dosage 15 mq / kq / adm SC

[0442] Individual serum IgA concentrations expressed in mg / mL and as percent-changes from baseline are reported in Table 9 for males and in Table 10 for females, respectively. Individual IgA concentrations across the study (from Day 1, predose to Day 36) ranged from 0.10 to 0.92 mg / mL with a mean and SD values of 0.42 ±0.25 mg / mL for males and from 0.10 to 1.40 mg / mL with a mean and SD values of 0.78 ±0.35 mg / mL for females, respectively. All animals showed reductions in IgA concentrations up to -88% (Female No. 17 on Day 36) occurring from Day 15 with a greater magnitude of change observed later in the dosing period at Day 36.Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP Table 9- BCMA NKCE (SAR445514) Dosage 15 mg / kg / adm SC- Males: Serum IgA Levels on Study Days

[0443]

[0444] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP Table 10- BCMA NKCE (SAR445514) Dosage 15 mg / kg / adm SC- Females: Serum IgA Levels on Study Days

[0445]

[0446] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP iv) BCMA NKCE (SAR445514) / Dosage 49.1 mg / kg / adm SC

[0447] Individual serum IgA levels expressed in mg / mL and as percent-changes from baseline are reported in Table 11 for males and in Table 12 for females, respectively. Individual IgA levels across the study (from Day 1 , predose to Day 36) ranged from 0.10 to 2.22 mg / mL with a mean and SD values of 0.99 ±0.75 mg / mL for males and from 0.46 to 2.36 mg / mL with a mean and SD values of 1.06 ±0.50 mg / mL for females, respectively. All animals showed reductions in IgA levels up to -96% (Male No. 20 on Day 36) occurring from Day 8 for 5 / 6 animals and later at Day 29 in Male No. 21 with a greater magnitude of change observed later in the dosing period at Day 36.

[0448] Table 11- BCMA NKCE (SAR445514) Dosage 49.1 mg / kg / adm SC- Males:

[0449] Serum IgA Levels on Study Days

[0450]

[0451] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0452]

[0453] Table 12- BCMA NKCE (SAR445514) Dosage 49.1 mg / kg / adm SC- Females:

[0454] Serum IgA Levels on Study Days

[0455]

[0456] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0457]

[0458] v) BCMA NKCE (SAR445514) / Dosage 30 mg / kg / adm IV

[0459] Individual serum IgA levels expressed in mg / mL and as percent-changes from baseline are reported in Table 13 for males and in Table 14 for females, respectively. Individual IgA levels across the study (from Day 1 , predose to Day 36) ranged from 0.10 to 1.24 mg / mL with a mean and SD values of 0.72 ±0.33 mg / mL for males and from 0.10 to 1.82 mg / mL with a mean and SD values of 0.69 ±0.43 mg / mL for females, respectively.

[0460] All animals showed reductions in IgA levels up to -90% (Female No. 28 on Day 36) occurring from Day 8 for one animal (Male No. 26) and Day 15 for other animals with a greater magnitude of change observed later in the dosing period at Day 36, except for one isolated animal (Male No. 27) where IgA levels were maintained during the study in the biological variation range.

[0461] Table 13- BCMA NKCE (SAR445514) Dosage 30 mg / kg / adm IV- Males: Serum IgA Levels on Study Days

[0462]

[0463] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0464]

[0465] Table 14- BCMA NKCE (SAR445514) Dosage 30 mg / kg / adm IV- Females: Serum IgA Levels on Study Days

[0466]

[0467] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0468]

[0469] vi) Conclusion (IgA Levels)

[0470] The data indicates that time-dependent reduction in serum IgA levels was observed following administration of BCMA NKCE (SAR445514). Reduction in serum IgA levels occurred at 5 mg / kg / adm SC to 49.1 mg / kg / adm SC and at 30 mg / kg / adm IV from Day 8 to Day 22 (depending on the animal), without a dose relationship to the SC route, and with the greatest magnitude of change observed later in the dosing period at Day 36.

[0471] IgG Levels

[0472] Baseline IgG levels observed on Day 1 , predose in both males and females reported in Table 4 ranged from 3.84 mg / mL to 8.15 mg / mL with a mean and SD values of 5.73 ±1.08 mg / mL.

[0473] i) BCMA NKCE (SAR445514) / Control Group

[0474] Individual serum IgG levels expressed in mg / mL and as percent-changes from baseline are reported in Table 15 for males and in Table 16 for females, respectively. Individual IgG levels across the study (from Day 1 , predose to Day 36)Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP ranged from 4.35 to 8.03 mg / mL with a mean and SD values of 6.18 ±0.95 mg / mL for males and from 5.32 to 8.58 mg / mL with a mean and SD values of 6.81 ±0.81 mg / mL for females, respectively.

[0475] Table 15- Control Group Males: Serum IgG levels on Study Days

[0476]

[0477] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0478] Table 16- Control Group Females: Serum IgG levels on Study Days

[0479]

[0480] ii) BCMA NKCE (SAR445514) / Dosage 5 mg / kg / adm SC

[0481] Individual serum IgG levels expressed in mg / mL and as percent-changes from baseline are reported in Table 17 for males and in Table 18 for females, respectively. Individual IgG levels across the study (from Day 1 , predose to Day 36)Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP ranged from 3.58 to 9.43 mg / mL with a mean and SD values of 5.57 ±1.54 mg / mL for males and from 4.02 to 9.49 mg / mL with a mean and SD values of 6.00 ±1.56 mg / mL for females, respectively. Increases in IgG levels (up to +107% in Female No. 11 on Day 29) were observed in 3 / 6 animals occurring from Day 22 in Female No. 11 and Day 29 in Males Nos. 8 and 9, respectively. No changes in IgG levels were observed during the study in the other 3 animals where changes were considered within the biological variation range.

[0482] Table 17- BCMA NKCE (SAR445514) Dosage 5 mg / kg / adm SC- Males: Serum IgG Levels on Study Days

[0483]

[0484] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0485]

[0486] Table 18- BCMA NKCE (SAR445514) Dosage 5 mg / kg / adm SC- Females: Serum IgG Levels on Study Days

[0487]

[0488] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0489]

[0490] Hi) BCMA NKCE (SAR445514) / Dosage 15 mg / kg / adm SC

[0491] Individual serum IgG levels expressed in mg / mL and as percent-changes from baseline are reported in Table 19 for males and in Table 20 for females, respectively. Individual IgG levels across the study (from Day 1 , predose to Day 36) ranged from 3.05 to 8.11 mg / mL with a mean and SD values of 5.34 ±1.39 mg / mL for males and from 4.61 to 10.59 mg / mL with a mean and SD values of 6.66 ±1.57 mg / mL for females, respectively. No changes in IgG levels were observed during the study in any animals except in one isolated animal (Female No. 18) showing increases in IgG levels with a maximal change of +129% observed at Day 29.

[0492] Table 19- BCMA NKCE (SAR445514) Dosage 15 mg / kg / adm SC- Males:

[0493] Serum IgG Levels on Study Days

[0494]

[0495] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0496]

[0497] Table 20- BCMA NKCE (SAR445514) Dosage 15 mg / kg / adm SC- Females: Serum IgG Levels on Study Days

[0498]

[0499] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0500]

[0501] iv) BCMA NKCE (SAR445514) / Dosage 49.1 mg / kg / adm SC

[0502] Individual serum IgG levels expressed in mg / mL and as percent-changes from baseline are reported in Table 21 for males and in Table 22 for females, respectively. Individual IgG levels across the study (from Day 1 , predose to Day 36) ranged from 2.26 to 10.21 mg / mL with a mean and SD values of 5.44 ±2.05 mg / mL for males and from 3.63 to 6.68 mg / mL with a mean and SD values of 5.03 ±0.94 mg / mL for females, respectively. Heterogenous responses were observed among the 6 animals. Increase in IgG levels (+80%) was observed in 1 / 6 animals (Male No.

[0503] 19) occurring from Day 29 to Day 36. Reduction in IgG levels (up to -54% on Day 36) was noted for Male No. 20. No changes in IgG levels were observed during the study in the other animals where changes were considered within the biological variation range.

[0504] Table 21- BCMA NKCE (SAR445514) Dosage 49.1 mg / kg / adm SC- Males:

[0505] Serum IgG Levels on Study Days

[0506]

[0507] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0508]

[0509] Table 22- BCMA NKCE (SAR445514) Dosage 49.1 mg / kg / adm SC- Females: Serum IgG Levels on Study Days

[0510]

[0511] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0512]

[0513] v) BCMA NKCE (SAR445514) / Dosage 30 mg / kg / adm IV

[0514] Individual serum IgG levels expressed in mg / mL and as percent-changes from baseline are reported in Table 23 for males and in Table 24 for females, respectively. Individual IgG levels across the study (from Day 1 , predose to Day 36) ranged from 2.94 to 14.28 mg / mL with a mean and SD values of 7.40 ±3.53 mg / mL for males and from 3.70 to 7.31 mg / mL with a mean and SD values of 5.17 ±1.04 mg / mL for females, respectively. Increases in IgG levels (up to +198% in Male No. 25 on Day 29) were observed in 4 / 6 animals (Males Nos. 25, 26, and 27, and Female No. 30) from Day 15 or Day 22. No changes in IgG levels were observed during the study in the 2 other animals where changes were considered within the biological variation range.Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP Table 23- BCMA NKCE (SAR445514) Dosage 30 mg / kg / adm IV- Males: Serum IgG Levels on Study Days

[0515]

[0516] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0517] Table 24- BCMA NKCE (SAR445514) Dosage 30 mg / kg / adm IV- Females: Serum IgG Levels on Study Days

[0518]

[0519] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP vi) Conclusion (IQG Levels)

[0520] No clear BCMA-NKCE (SAR445514)-related effect on serum IgG levels was evidenced. Heterogenous responses were observed among all animals treated with BCMA-NKCE (SAR445514). When changes were observed, they consisted of either increased IgG levels occurring from 5 mg / kg / adm SC and from Day 15 to Day 36 (depending on the animal), or reduction of IgG levels occurring from Day 22 (observed only at 49.1 mg / kg / adm SC in one isolated animal) with a greater magnitude of change observed at Day 36.

[0521] IQM Levels

[0522] Baseline IgM levels observed on Day 1 , predose in both males and females reported in Table 4 ranged from 0.73 mg / mL to 2.85 mg / mL with a mean and SD values of 1.45 ±0.50 mg / mL.

[0523] / ') BCMA NKCE (SAR445514) / Control Group

[0524] Individual serum IgM levels expressed in mg / mL and as percent-changes from baseline are reported in Table 25 for males and in Table 26 for females, respectively. Individual IgM levels across the study (from Day 1 , predose to Day 36) ranged from 1.01 to 2.09 mg / mL with a mean and SD values of 1.42 ±0.32 mg / mL for males and from 0.96 to 2.22 mg / mL with a mean and SD values of 1.47 ±0.41 mg / mL for females, respectively.

[0525] Table 25- Control Group Males: Serum IgM levels on Study Days

[0526]

[0527] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0528]

[0529] Table 26- Control Group Females: Serum IgM levels on Study Days

[0530]

[0531] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0532]

[0533] ii) BCMA NKCE (SAR445514) / Dosage 5 mq / kq / adm SC

[0534] Individual serum IgM levels expressed in mg / mL and as percent-changes from baseline are reported in Table 27 for males and in Table 28 for females, respectively. Individual IgM levels across the study (from Day 1 , predose to Day 36) ranged from 0.30 to 1.51 mg / mL with a mean and SD values of 0.89 ±0.36 mg / mL and from 0.38 to 2.85 mg / mL with a mean and SD values of 1.41 ±0.66 mg / mL for females, respectively. 5 / 6 animals (Males Nos. 7, 8 and 9, and Females Nos. 10 and 12) showed reductions in IgM levels (up to -74% in Male No. 7 on Day 36) occurring from Day 2 or Day 15 with a greater magnitude of change observed later in the dosing period at Day 36 in Male No. 7 and Female No. 10 whereas for Males Nos. 8 and 9, a return close to the baseline value is observed at Day 36. No changes in IgM levels were observed during the study in 1 / 6 animals (Female No. 11 ) where changes were considered within the biological variation range.Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP Table 27- BCMA NKCE (SAR445514) Dosage 5 mg / kg / adm SC- Males: Serum IgM Levels on Study Days

[0535]

[0536] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP Table 28- BCMA NKCE (SAR445514) Dosage 5 mg / kg / adm SC- Females:

[0537] Serum IgM Levels on Study Days

[0538]

[0539] Hi) BCMA NKCE (SAR445514) / Dosage 15 mg / kg / adm SC

[0540] Individual serum IgM levels expressed in mg / mL and as percent-changes from baseline are reported in Table 29 for males and in Table 30 for females, respectively. Individual IgM levels across the study (from Day 1 , predose to Day 36) ranged from 0.29 to 2.58 mg / mL with a mean and SD values of 1.04 ±0.64 mg / mL for males and from 0.37 to 1.71 mg / mL with a mean and SD values of 0.94 ±0.37Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP mg / mL for females, respectively. 5 / 6 animals showed reductions in IgM levels (up to -81% in Male No. 13 on Day 36) occurring from Day 8 with a greater magnitude of change observed later in the dosing period at Day 29 or Day 36 except for one isolated animal (Female No. 18) where IgM levels was maintained throughout the study in the biological variation range.

[0541] Table 29- BCMA NKCE (SAR445514) Dosage 15 mg / kg / adm SC- Males:

[0542] Serum IgM Levels on Study Days

[0543]

[0544] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0545]

[0546] Table 30- BCMA NKCE (SAR445514) Dosage 15 mg / kg / adm SC- Females:

[0547] Serum IgM Levels on Study Days

[0548]

[0549] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP iv) BCMA NKCE (SAR445514) / Dosage 49.1 mg / kg / adm SC

[0550] Individual serum IgM levels expressed in mg / mL and as percent-changes from baseline are reported in Table 31 for males and in Table 32 for females, respectively. Individual IgM levels across the study (from Day 1 , predose to Day 36) ranged from 0.10 to 0.95 mg / mL with a mean and SD values of 0.53 ±0.25 mg / mL for males and from 0.22 to 2.21 mg / mL with a mean and SD values of 0.94 ±0.62 mg / mL for females, respectively. All animals showed reductions in IgM levels (up to -90% in Male No. 20 on Day 22) occurring from Day 8 or Day 15 with a greater magnitude of change observed later in the dosing period at Day 36.

[0551] Table 31- BCMA NKCE (SAR445514) Dosage 49.1 mg / kg / adm SC- Males:

[0552] Serum IgM Levels on Study Days

[0553]

[0554] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0555]

[0556] Table 32- BCMA NKCE (SAR445514) Dosage 49.1 mg / kg / adm SC- Females: Serum IgM Levels on Study Days

[0557]

[0558] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0559]

[0560] v) BCMA NKCE (SAR445514) / Dosage 30 mg / kg / adm IV

[0561] Individual serum IgM levels expressed in mg / mL and as percent-changes from baseline are reported in Table 33 for males and in Table 34 for females, respectively. Individual IgM levels across the study (from Day 1 , predose to Day 36) ranged from 0.32 to 1.21 mg / mL with a mean and SD values of 0.72 ±0.22 mg / mL for males and from 0.33 to 2.16 mg / mL with a mean and SD values of 0.93 ±0.53 mg / mL for females, respectively. All animals showed reductions in IgM levels up to -70% occurring from Day 8 or Day 15 with a greater magnitude of change observed later in the dosing period at Day 36.

[0562] Table 33- BCMA NKCE (SAR445514) Dosage 30 mg / kg / adm IV- Males: Serum IgM Levels on Study Days

[0563]

[0564] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0565]

[0566] Table 34- BCMA NKCE (SAR445514) Dosage 30 mg / kg / adm IV- Females: Serum IgM Levels on Study Days

[0567]

[0568] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0569]

[0570] vi) Conclusion (IgM Levels)

[0571] The data indicates that time-dependent reduction in serum IgM levels was observed following administration of BCMA NKCE (SAR445514). Reduction in serum IgM levels occurred at 5 mg / kg / adm SC to 49.1 mg / kg / adm SC and at 30 mg / kg / adm IV from Day 2 to Day 15 (depending on the animal), without a dose relationship to the SC route, and with the greatest magnitude of change observed later in the dosing period at Day 36.

[0572] Conclusion:

[0573] This study demonstrates that BCMA NKCE (SAR445514) induced a timedependent reduction in serum IgA and IgM levels without any clear effect on serum IgG levels. The reduction of serum IgA and IgM levels occurred 7 days post first dosing at 5 mg / kg / adm SC to 49.1 mg / kg / adm SC and at 30 mg / kg / adm IV, without a dose relationship to the SC route, and with the greatest magnitude of change observed on Day 36.Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP SEQUENCE LISTING

[0574]

[0575] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0576]

[0577] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0578]

[0579] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0580]

[0581] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0582]

[0583] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0584]

[0585] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0586]

[0587] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0588]

[0589] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0590]

[0591] Attorney Ref.: 776083: SA9-513PC Sanofi Ref: PAT25048-WO1-NP

[0592]

Claims

Attorney Ref.: 762816: SA9-513EP-1Sanofi Ref: PAT25048CLAIMS1. A method of treating an autoimmune disorder in a subject, comprising administering to the subject a binding protein comprising a first antigen binding domain (ABD) with binding specificity to BCMA and a second ABD with binding specificity to NKp46, thereby treating the B-cell mediated autoimmune disorder in the subject.

2. The method of claim 1 , wherein:(a) the first ABD comprises:(a1 ) a first immunoglobulin heavy chain variable domain (VH1 ) comprising an HCDR1 comprising the amino acid sequence GFTFSNFGMH (SEQ ID NO: 1), an HCDR2 comprising the amino acid sequence VIWSDETNR (SEQ ID NO: 2), and an HCDR3 comprising the amino acid sequence DQQYCSSDSCFTWFDP (SEQ ID NO: 3); and(a2) a first immunoglobulin light chain variable domain (VL1) comprising an LCDR1 comprising the amino acid sequence CASSTGTVTPSNYAN (SEQ ID NO: 7), an LCDR2 comprising the amino acid sequence DNNSRPP (SEQ ID NO: 8), and an LCDR3 comprising the amino acid sequence ALWFGNQWV (SEQ ID NO: 9); or(a) the first ABD comprises:(a1 ) a first immunoglobulin heavy chain variable domain (VH1 ) comprising an HCDR1 comprising the amino acid sequence GFTFSNFG (SEQ ID NO: 112), an HCDR2 comprising the amino acid sequence IWSDETNR (SEQ ID NO: 113), and an HCDR3 comprising the amino acid sequence ARDQQYCSSDSCFTWFDP (SEQ ID NO: 114); and(a2) a first immunoglobulin light chain variable domain (VL1) comprising an LCDR1 comprising the amino acid sequence TGTVTPSNY (SEQ ID NO: 118), an LCDR2 comprising the amino acid sequence DNN (SEQ ID NO: 119), and an LCDR3 comprising the amino acid sequence ALWFGNQWV (SEQ ID NO: 120); or(a) the first ABD comprises:Attorney Ref.: 762816: SA9-513EP-1Sanofi Ref: PAT25048 (a1 ) a first immunoglobulin heavy chain variable domain (VH1 ) comprising an HCDR1 comprising the amino acid sequence NFGMH (SEQ ID NO: 115), an HCDR2 comprising the amino acid sequence VIWSDETNRYYADSVKG (SEQ ID NO: 116), and an HCDR3 comprising the amino acid sequence DQQYCSSDSCFTWFDP (SEQ ID NO: 117); and(a2) a first immunoglobulin light chain variable domain (VL1) comprising an LCDR1 comprising the amino acid sequence ASSTGTVTPSNYAN (SEQ ID NO: 121), an LCDR2 comprising the amino acid sequence DNNSRPP (SEQ ID NO: 122), and an LCDR3 comprising the amino acid sequence ALWFGNQWV (SEQ ID NO: 123).

3. The method of claim 1 or 2, wherein (b) the second ABD comprises: (b1) a second immunoglobulin heavy chain variable domain (VH2) comprising:- an HCDR1 comprising the amino acid sequence DYVIN, an HCDR2 comprising the amino acid sequence EIYPGSGTNYYNEKFKA, and an HCDR3 comprising the amino acid sequence RGRYGLYAMDY;- an HCDR1 sequence comprising GYTFSDYVIN (SEQ ID NO: 19), an HCDR2 sequence comprising EIYPGSGTN (SEQ ID NO: 20), and an HCDR3 sequence comprising RGRYGLYAMDY (SEQ ID NO: 21);- an HCDR1 comprising the amino acid sequence SDYAWN (SEQ ID NO: 22), an HCDR2 comprising the amino acid sequence YITYSGSTSYNPSLES (SEQ ID NO: 23), and an HCDR3 comprising the amino acid sequence GGYYGSSWGVFAY (SEQ ID NO: 24);- an HCDR1 comprising the amino acid sequence EYTMH (SEQ ID NO: 25), an HCDR2 comprising the amino acid sequence GISPNIGGTSYNQKFKG (SEQ ID NO: 26), and an HCDR3 comprising the amino acid sequence RGGSFDY (SEQ ID NO: 27);- an HCDR1 comprising the amino acid sequence SFTMH (SEQ ID NO: 28), an HCDR2 comprising the amino acid sequence YINPSSGYTEYNQKFKD (SEQ ID NO: 29), and an HCDR3 comprising the amino acid sequence GSSRGFDY (SEQ ID NO: 30); orAttorney Ref.: 762816: SA9-513EP-1Sanofi Ref: PAT25048 - an HCDR1 comprising the amino acid sequence SDYAWN (SEQ ID NO: 31 ), an HCDR2 comprising the amino acid sequence YITYSGSTNYNPSLKS (SEQ ID NO: 32), and an HCDR3 comprising the amino acid sequence CWDYALYAMDC (SEQ ID NO: 33); and(b2) a second immunoglobulin light chain variable domain (VL2) comprising:- an LCDR1 comprising the amino acid sequence RASQDISNYLN (SEQ ID NO: 34), an LCDR2 comprising the amino acid sequence YTSRLHS (SEQ ID NO: 35), and an LCDR3 comprising the amino acid sequence QQGNTRPWT (SEQ ID NO: 36);- an LCDR1 comprising the amino acid sequence RVSENIYSYLA (SEQ ID NO: 37), an LCDR2 comprising the amino acid sequence NAKTLAE (SEQ ID NO: 38), and an LCDR3 comprising the amino acid sequence QHHYGTPWT (SEQ ID NO: 39);- an LCDR1 comprising the amino acid sequence RASQSISDYLH (SEQ ID NO: 40), an LCDR2 comprising the amino acid sequence YASQSIS (SEQ ID NO: 41 ), and an LCDR3 comprising the amino acid sequence QNGHSFPLT (SEQ ID NO: 42);- an LCDR1 comprising the amino acid sequence RASENIYSNLA (SEQ ID NO: 43), an LCDR2 comprising the amino acid sequence AATN LAD (SEQ ID NO: 44), and an LCDR3 comprising the amino acid sequence QHFWGTPRT (SEQ ID NO: 45); or- an LCDR1 comprising the amino acid sequence RTSENIYSYLA (SEQ ID NO: 46), an LCDR2 comprising the amino acid sequence NAKTLAE (SEQ ID NO: 47), and an LCDR3 comprising the amino acid sequence QHHYDTPLT (SEQ ID NO: 48).

4. The method of claim 2 or 3, wherein the VL1 comprises:- an LCDR1 sequence comprising the amino acid sequence of CASSTGTVTPSNYAN (SEQ ID NO: 7), an LCDR2 sequence comprising the amino acid sequence of DNNSRPP (SEQ ID NO: 8), and an LCDR3 sequence comprising the amino acid sequence of ALWFGNQWV (SEQ ID NO: 9);- an LCDR1 sequence comprising the amino acid sequence of CRSSTGTVTPSNYAN (SEQ ID NO: 10), an LCDR2 sequence comprising the aminoAttorney Ref.: 762816: SA9-513EP-1Sanofi Ref: PAT25048 acid sequence of DNNSRPP (SEQ ID NO: 11), and an LCDR3 sequence comprising the amino acid sequence of ALWFGNQWV (SEQ ID NO: 12);- an LCDR1 sequence comprising the amino acid sequence of CASSTGAVTPSNYAN (SEQ ID NO: 13), an LCDR2 sequence comprising the amino acid sequence of DNNIKPP (SEQ ID NO: 14), and an LCDR3 sequence comprising the amino acid sequence of ALWYGGQVW (SEQ ID NO: 15); or- an LCDR1 sequence comprising the amino acid sequence of CASSTGAVTPGYYAN (SEQ ID NO: 16), an LCDR2 sequence comprising the amino acid sequence of DNNNKPP (SEQ ID NO: 17), and an LCDR3 sequence comprising the amino acid sequence of ALYYGGQWV (SEQ ID NO: 18).

5. The method of claim 1 , wherein:(a) the first ABD comprises:(a1 ) a first immunoglobulin heavy chain variable domain (VH1 ) comprising an HCDR1 sequence comprising the amino acid sequence of GFTFSNFG (SEQ ID NO: 112), an HCDR2 sequence comprising the amino acid sequence of IWSDETNR (SEQ ID NO: 113), and an HCDR3 sequence comprising the amino acid sequence of ARDQQYCSSDSCFTWFDP (SEQ ID NO: 114); and(a2) a first immunoglobulin light chain variable domain (VL1) comprising an LCDR1 sequence comprising the amino acid sequence of TGTVTPSNY (SEQ ID NO: 118), an LCDR2 sequence comprising the amino acid sequence of DNN (SEQ ID NO: 119), and an LCDR3 sequence comprising the amino acid sequence of ALWFGNQWV (SEQ ID NO: 120).

6. The method of claim 1 , wherein:(a) the first ABD comprises:(a1 ) a first immunoglobulin heavy chain variable domain (VH1 ) comprising an HCDR1 sequence comprising the amino acid sequence of NFGMH (SEQ ID NO: 115), an HCDR2 sequence comprising the amino acid sequence of VIWSDETNRYYADSVKG (SEQ ID NO: 116), and an HCDR3 sequence comprising the amino acid sequence of DQQYCSSDSCFTWFDP (SEQ ID NO: 117); andAttorney Ref.: 762816: SA9-513EP-1Sanofi Ref: PAT25048 (a2) a first immunoglobulin light chain variable domain (VL1) comprising an LCDR1 sequence comprising the amino acid sequence of ASSTGTVTPSNYAN (SEQ ID NO: 121), an LCDR2 sequence comprising the amino acid sequence of DNNSRPP (SEQ ID NO: 122), and an LCDR3 sequence comprising the amino acid sequence of ALWFGNQWV (SEQ ID NO: 123).

7. The method of any one of claims 2-6, wherein:- the VH1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 55;- the VH1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 50;- the VH1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 51 ;- the VH1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 52;- the VH1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 53; or- the VH1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 54.Attorney Ref.: 762816: SA9-513EP-1Sanofi Ref: PAT25048 8. The method of any one of claims 2-7, wherein:- the VH1 comprises an amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence of SEQ ID NO: 55;- the VH1 comprises an amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence of SEQ ID NO: 50;- the VH1 comprises an amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence of SEQ ID NO: 51 ;- the VH1 comprises an amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence of SEQ ID NO: 52;- the VH1 comprises an amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence of SEQ ID NO: 53; or- the VH1 comprises an amino acid sequence of SEQ ID NO: 49, and wherein the VL1 comprises an amino acid sequence of SEQ ID NO: 54.

9. The method of any one of claims 3-8, wherein:- the VH2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 56, and wherein the VL2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 64;- the VH2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 57, and wherein the VL2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 65;- the VH2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 58, and wherein the VL2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 66;- the VH2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 59, and wherein the VL2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 67;Attorney Ref.: 762816: SA9-513EP-1Sanofi Ref: PAT25048 - the VH2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 60, and wherein the VL2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 68;- the VH2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 61, and wherein the VL2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 69;- the VH2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 62, and wherein the VL2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 70; or- the VH2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 63, and wherein the VL2 comprises an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 71.

10. The method of any one of claims 3-9, wherein:- the VH2 comprises an amino acid sequence of SEQ ID NO: 56, and wherein the VL2 comprises an amino acid sequence of SEQ ID NO: 64;- the VH2 comprises an amino acid sequence of SEQ ID NO: 57, and wherein the VL2 comprises an amino acid sequence of SEQ ID NO: 65;- the VH2 comprises an amino acid sequence of SEQ ID NO: 58, and wherein the VL2 comprises an amino acid sequence of SEQ ID NO: 66;- the VH2 comprises an amino acid sequence of SEQ ID NO: 59, and wherein the VL2 comprises an amino acid sequence of SEQ ID NO: 67;- the VH2 comprises an amino acid sequence of SEQ ID NO: 60, and wherein the VL2 comprises an amino acid sequence of SEQ ID NO: 68;- the VH2 comprises an amino acid sequence of SEQ ID NO: 61, and wherein the VL2 comprises an amino acid sequence of SEQ ID NO: 69;- the VH2 comprises an amino acid sequence of SEQ ID NO: 62, and wherein the VL2 comprises an amino acid sequence of SEQ ID NO: 70; orAttorney Ref.: 762816: SA9-513EP-1Sanofi Ref: PAT25048 - the VH2 comprises an amino acid sequence of SEQ ID NO: 63, and wherein the VL2 comprises an amino acid sequence of SEQ ID NO: 71.

11. The method of any one of claims 1 -10, wherein the binding protein further comprises all or part of an immunoglobulin Fc domain or variant thereof.

12. The method of claim 11, wherein all or part of the immunoglobulin Fc domain or variant thereof binds to a human Fc-y receptor.

13. The method of claim 11 or 12, wherein all or part of the immunoglobulin Fc domain or variant thereof binds to a human CD16A (FcyRIII) polypeptide.

14. The method of any one of claims 11-13, wherein the Fc domain comprises a native glycan at amino acid position 297, according to Ell numbering.

15. The method of any one of claims 11-14, wherein the binding protein is N-glycosylated.

16. The method of any one of claims 11-15, wherein the Fc domain or variant thereof comprises a first Fc heavy chain and a second Fc heavy chain.

17. The method of claim 16, wherein at least one Fc heavy chain comprises an engineered intrachain disulfide bond mediated by a pair of cysteines (C) that substitute for:(i) a leucine (L) at amino acid position 242 and a lysine (K) at amino acid position 334;or(ii) an arginine (R) at amino acid position 292 and a valine (V) at amino acid position 302;wherein the amino acid positions are according to Ell numbering.Attorney Ref.: 762816: SA9-513EP-1Sanofi Ref: PAT25048 18. The method of claim 17, wherein the first and the second Fc heavy chain each comprise both the L242C and K334C substitutions.

19. The method of claim 17, wherein the first and the second Fc heavy chain each comprise both the R292C and V302C substitutions.

20. The method of any one of claims 16-19, wherein at least one Fc heavy chain comprises a substitution at amino acid position 332, according to Ell numbering.

21. The method of claim 20, wherein the substitution at amino acid position 332 is a glutamic acid (E).

22. The method of claim 20 or 21 , wherein the at least one Fc heavy chain further comprises one or more substitutions at amino acid positions 236, 239, or 330, according to Ell numbering.

23. The method of claim 22, wherein the substitution at amino acid position 236 is an alanine (A), the substitution at amino acid position 239 is an aspartic acid (D), and the substitution at amino acid position 330 is a leucine (L).

24. The method of any one of claims 16-23, wherein at least one Fc heavy chain comprises an aspartic acid (D) at amino acid position 239, and a glutamic acid (E) at amino acid position 332, according to Ell numbering.

25. The method of any one of claims 16-24, wherein at least one Fc heavy chain comprises an alanine (A) at amino acid position 236, an aspartic acid (D) at amino acid position 239, and a glutamic acid (E) at amino acid position 332, according to Ell numbering.

26. The method of any one of claims 16-25, wherein at least one Fc heavy chain further comprises an alanine (A) at amino acid position 236, an aspartic acid (D)Attorney Ref.: 762816: SA9-513EP-1Sanofi Ref: PAT25048 at amino acid position 239, a leucine (L) at amino acid position 330, and a glutamic acid (E) at amino acid position 332, according to Ell numbering.

27. The method of any one of claims 3-26, wherein the binding protein comprising at least two polypeptide chains that form at least two antigen-binding sites, wherein at least one polypeptide chain comprises a structure represented by the formula:VL1-L1-VL2-L2-CL [I];and at least one polypeptide chain comprises a structure represented by the formula:VH2-L3-VH1-L4-CH1 [II];wherein:CL is an immunoglobulin light chain constant domain;CH1 is an immunoglobulin CH1 heavy chain constant domain; and L1 , L2, L3, and L4 are amino acid linkers, wherein any one or more of L1 , L2, L3, and L4 are optionally absent, andwherein the polypeptides of formula I and the polypeptides of formula II form a cross-over light chain-heavy chain pair.

28. The method of any one of claims 3-27, wherein the binding protein comprises three polypeptide chains that form two antigen-binding sites, wherein one polypeptide chain comprises a structure represented by the formula:VL1-L1-VL2-L2-CL [I];one polypeptide chain comprises a structure represented by the formula: VH2-L3-VH1-L4-CH1-hinge-CH2-CH3 [III]; andone polypeptide chain comprises a structure represented by the formula: hinge-CH2-CH3 [IV]wherein:Attorney Ref.: 762816: SA9-513EP-1Sanofi Ref: PAT25048 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;andL1 , L2, L3, and L4 are amino acid linkers, wherein any one or more of L1 , L2, L3, and L4 are optionally absent, andwherein the polypeptides of formula I and the polypeptides of formula II form a cross-over light chain-heavy chain pair.

29. The method of claim 27 or 28, wherein:(a) L1, L2, L3, and L4 each independently are zero amino acids in length or comprise a sequence selected from the group consisting of GGGGSGGGGS, GGGGSGGGGSGGGGS, S, RT, TKGPS, GQPKAAP, and GGSGSSGSGG; or (b) L1, L2, L3, and L4 each independently comprise a sequence selected from the group consisting of GGGGSGGGGS, GGGGSGGGGSGGGGS, S, RT, TKGPS, GQPKAAP, and GGSGSSGSGG.

30. The method of any one of claims 1-29, wherein:(a) the first ABD of the binding protein comprises a first immunoglobulin heavy chain variable domain (VH1 ) comprising an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 49, and a first immunoglobulin light chain variable domain (VL1 ) comprising an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 55; and (b) the second ABD of the binding protein comprises a second immunoglobulin heavy chain variable domain (VH2) comprising an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 56, and a second immunoglobulin light chain variable domain (VL2) comprising an amino acid sequence that is at least about 90% identical to the amino acid sequence of SEQ ID NO: 64.Attorney Ref.: 762816: SA9-513EP-1Sanofi Ref: PAT2504831. The method of any one of claims 1-30, wherein the binding protein comprises:(i) a first polypeptide chain comprising an amino acid sequence of SEQ ID NO: 72;(ii) a second polypeptide chain comprising an amino acid sequence of SEQ ID NO: 73; and(iii) a third polypeptide chain comprising an amino acid sequence of SEQ ID NO: 74.

32. The method of any one of claims 1 or 2, wherein:(b) the second ABD comprises:(b1) a second immunoglobulin heavy chain variable domain (VH2) comprising an HCDR1 sequence comprising the amino acid sequence of GYTFSDYV (SEQ ID NO: 87), an HCDR2 sequence comprising the amino acid sequence of IYPGSGTN (SEQ ID NO: 90), and an HCDR3 sequence comprising the amino acid sequence of ARRGRYGLYAMDY (SEQ ID NO: 93); and(b2) a second immunoglobulin light chain variable domain (VL1 ) comprising an LCDR1 sequence comprising the amino acid sequence of QDISNY (SEQ ID NO: 96), an LCDR2 sequence comprising the amino acid sequence of YTS (SEQ ID NO: 99), and an LCDR3 sequence comprising the amino acid sequence of QQGNTRPWT (SEQ ID NO: 111).

33. The method of any one of claims 1 or 2, wherein:(b) the second ABD comprises:(b1) a second immunoglobulin heavy chain variable domain (VH2) comprising an HCDR1 sequence comprising the amino acid sequence of DYVIN (SEQ ID NO: 80), an HCDR2 sequence comprising the amino acid sequence of EIYPGSGTNYYNEKFKA (SEQ ID NO: 81), and an HCDR3 sequence comprising the amino acid sequence of RGRYGLYAMDY (SEQ ID NO: 21); and(b2) a second immunoglobulin light chain variable domain (VL1 ) comprising an LCDR1 sequence comprising the amino acid sequence of RASQDISNYLN (SEQAttorney Ref.: 762816: SA9-513EP-1Sanofi Ref: PAT25048 ID NO: 34), an LCDR2 sequence comprising the amino acid sequence of YTSRLHS (SEQ ID NO: 35), and an LCDR3 sequence comprising the amino acid sequence of QQGNTRPWT (SEQ ID NO: 36).

34. The method of any one of claims 1-33, wherein the autoimmune disorder is a B-cell mediated autoimmune disorder.

35. The method of any one of claims 1-33, wherein the autoimmune disorder is a plasma cell mediated autoimmune disorder.

36. The method of any one of claims 1-35, wherein the autoimmune disorder is lupus.

37. The method of claim 36, wherein the lupus is systemic lupus erythematosus (SLE).

38. The method of any one of claims 1-34, wherein the autoimmune disorder is rheumatoid arthritis (RA).

39. The method of any one of claims 1-34, wherein the subject has a reduced serum immunoglobulin level compared to a subject that was not administered the binding protein.

40. The method of any one of claims 1-34, wherein subject has a reduced autoantibody level compared to a subject that was not administered the binding protein.

41. The method of claim 39 or 40, wherein the serum immunoglobulin and / or auntoantibody is IgA, IgM, and / or IgG.Attorney Ref.: 762816: SA9-513EP-1Sanofi Ref: PAT25048 42. The method of any one of claims 1-41, wherein the subject has a reduced serum rheumatoid factor (RF) level compared to a subject that was not administered the binding protein.

43. The method of any one of claims 1-42, wherein the binding protein is administered intravenously or subcutaneously.

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