Methods for reducing serum levels of Fc-containing agents using FcRn antagonists

Abstract

To provide pharmaceutical compositions for use in methods of reducing the serum levels of Fc-containing agents (e.g., antibodies and immunoadhesins) in a subject.SOLUTION: The present invention provides pharmaceutical compositions for use in methods of reducing the serum levels of autoantibodies in a subject, wherein it comprises a FcRn antagonist comprising a variant Fc region, at least one Fc domain in the variant Fc region comprises a specific amino acid, the FcRn-antagonist is administered to the subject in a dose of between about 2 and about 200 mg / kg and the FcRn-antagonist is administered to the subject at least twice in 20 days.SELECTED DRAWING: None

Description

[Technical Field]

[0001] (Related Applications) This application is incorporated herein by reference in its entirety. The benefit of Application No. 62 / 130,076 is claimed. [Background technology]

[0002] (background) Immunoglobulin gamma (IgG) antibodies are used to treat autoimmune and inflammatory diseases, and their pathology is related to IgG antibodies. Many disorders, such as those characterized by excessive expression of plays an important role in the pathology of cancer (see, e.g., Junghans, Immunologic Research 16 (1):29 (1997).

[0003] The half-life of IgG in serum is prolonged compared to the serum half-lives of other plasma proteins (Roope Nian et al., J. Immunology 170:3528 (2003); Junghans and Anderson, Proc. atl. Acad. Sci. USA 93:5512 (1996)). This long half-life is due, in part, to the Fc region of IgG. This is caused by the binding of FcRn, an Fc receptor, to IgG, which is inherited from the mother. Although FcRn was characterized as a fetal transport receptor for IgG, it appears that FcRn may degrade IgG in adults. FcRn also functions to protect the IgG taken up by pinocytosis from the immune system. Recycling back to the extracellular compartment prevents them from being transported to the lysosomes for degradation. This recycling is facilitated by the pH-dependent binding of IgG to FcRn. The IgG / FcRn interaction is more pronounced at the acidic intraendosomal pH than at extracellular physiological pH. It will be stronger than ever.

[0004] When the serum concentration of IgG reaches a level that exceeds the available FcRn molecules, unbound IgG It is no longer protected from degradation mechanisms and consequently has a reduced serum half-life. Therefore, inhibiting IgG binding to FcRn prevents IgG recycling through the endosomes. This reduces the serum half-life of IgG. Therefore, agents that antagonize IgG binding to FcRn modulates, treats, or inhibits antibody-mediated disorders such as autoimmune diseases, inflammatory diseases, and the like; One method for antagonizing IgG Fc binding to FcRn is to This involves the production of blocking antibodies against FcRn (see, for example, WO2002 / 43658). Peptides that bind to and antagonize the function of FcRn have also been identified (see, for example, US Pat. No. 6,212,022). and US 8,101,186). Furthermore, binding to FcRn is enhanced and pH dependent. Full-length IgG antibodies containing reduced variant Fc receptors also compete for FcRn binding to IgG However, antibody-mediated There is a need in the art for improved methods of treating disorders. Summary of the Invention

[0005] (overview) The present disclosure provides methods for measuring serum levels of Fc-containing agents (e.g., antibodies and immunoadhesins) in a subject. These methods generally involve administering to the subject a native Fc specifically binds to FcRn with increased affinity and reduced pH dependence compared to the FcRn domain, The disclosed methods include administering an effective amount of an isolated FcRn antagonist to a subject. They are particularly useful in the treatment of mediated disorders (eg, autoimmune diseases).

[0006] Thus, in one aspect, the present disclosure provides a method for reducing serum levels of an Fc-containing agent in a subject. administering to the subject an effective amount of an isolated antibody comprising a variant Fc region or an FcRn-binding fragment thereof and administering to the patient an FcRn antagonist having a nucleotide sequence selected from the group consisting of: ... containing amino acids Y, T, E, K, F, and Y at positions 252, 254, 256, 433, 434, and 436, respectively and the FcRn antagonist is administered to the subject at a dose of about 0.2 to about 200 mg / kg. Provide the law.

[0007] In another aspect, the present disclosure provides a method for reducing serum levels of an Fc-containing agent in a subject. administering to the subject an effective amount of an isolated antibody comprising a variant Fc region or an FcRn-binding fragment thereof administering an FcRn antagonist, wherein the Fc domain of the variant Fc region is at EU position 25 containing the amino acids Y, T, E, K, F, and Y at positions 2, 254, 256, 433, 434, and 436, respectively; and wherein the FcRn antagonist is administered to the subject at least twice every 20 days. .

[0008] The following embodiments apply to all aspects of this disclosure.

[0009] In certain embodiments, the FcRn antagonist is administered to the subject in the amount of 1, 2, 3, 4, 5, 6, 7, 8, In certain embodiments, the FcRn antagonist is administered once every 9 or 10 days. is administered to the subject once every four days. In certain embodiments, the FcRn antagonist is administered to the subject once every seven days. The strikes will be held on the 20th on the 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, and 18th of each month. , 19, or 20 doses.

[0010] In another aspect, the present disclosure provides a method for reducing serum levels of an Fc-containing agent in a subject. administering to the subject an effective amount of an isolated antibody comprising a variant Fc region or an FcRn-binding fragment thereof administering an FcRn antagonist, wherein the Fc domain of the variant Fc region is at EU position 25 containing the amino acids Y, T, E, K, F, and Y at positions 2, 254, 256, 433, 434, and 436, respectively; and The FcRn antagonist is administered to the subject once every 48 hours for four weeks. A method is provided.

[0011] In certain embodiments, the FcRn antagonist is administered to the subject at a dose of about 0.2 to about 200 mg / kg. In certain embodiments, the FcRn antagonist is administered to the subject in an amount of about 0.2, 1 , 2, 3, 5, 10, 20, 25, 30, 50, 70, 100, or 200 mg / kg. In an embodiment, the FcRn antagonist is administered to the subject at a dose of about 10 mg / kg. In certain embodiments, the FcRn antagonist is administered to the subject at a dose of about 20 mg / kg. In certain embodiments, the FcRn antagonist is administered to the subject at a dose of about 25 mg / kg. It is administered.

[0012] In another aspect, the present disclosure provides a method for reducing serum levels of an Fc-containing agent in a subject. administering to the subject an effective amount of an isolated antibody comprising a variant Fc region or an FcRn-binding fragment thereof administering an FcRn antagonist, wherein the Fc domain of the variant Fc region is at EU position 25 containing the amino acids Y, T, E, K, F, and Y at positions 2, 254, 256, 433, 434, and 436, respectively; and The FcRn antagonist is administered to the subject at a dose of about 25 mg / kg once every four days. A method is provided.

[0013] In another aspect, the present disclosure provides a method for reducing serum levels of an Fc-containing agent in a subject. administering to the subject an effective amount of an isolated antibody comprising a variant Fc region or an FcRn-binding fragment thereof administering an FcRn antagonist, wherein the Fc domain of the variant Fc region is at EU position 25 containing the amino acids Y, T, E, K, F, and Y at positions 2, 254, 256, 433, 434, and 436, respectively; and The FcRn antagonist is administered to the subject at a dose of about 25 mg / kg once every seven days. A method is provided.

[0014] In certain embodiments, the FcRn antagonist is administered intravenously. In certain embodiments, the FcRn antagonist is administered subcutaneously. The antagonist is administered to the subject in two or more doses, wherein the first dose administered to the subject is administered intravenously and the second or one or more subsequent doses are administered subcutaneously.

[0015] In certain embodiments, the FcRn antagonist does not comprise an antibody variable region. In certain embodiments, the FcRn antagonist does not comprise a CH1 domain. In certain embodiments, the FcRn antagonist does not contain a free cysteine ​​residue. In certain embodiments, the variant Fc region is an IgG Fc region. In certain embodiments, the variant Fc region is an IgG1 Fc region. The amino acid sequence comprises the amino acid sequence set forth in SEQ ID NO: 1, 2, or 3. wherein the amino acid sequence of the Fc domain of the variant Fc region is the amino acid sequence set forth in SEQ ID NO: 1. In certain embodiments, the FcRn antagonist comprises a variant Fc region. The amino acid sequence of the Fc domain of the variant Fc region is SEQ ID NO: 1, 2, or It consists of the amino acid sequence set forth in 3.

[0016] In certain embodiments, the variant Fc region binds to an Fcγ receptor of a wild-type IgG1 Fc region. In certain embodiments, the antibody has increased affinity for Fcγ receptors compared to the affinity for Fcγ receptors. In certain embodiments, the variant Fc region has increased affinity for CD16a. wherein the Fc domain of the variant Fc region does not contain an N-linked glycan at EU position 297. In certain embodiments, the FcRn antagonist comprises multiple FcRn antagonist molecules. and at least 50% (optionally, at least 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 29 , 90, 95, or 99%) of the variants F contain an afucosylated N-linked glycan at EU position 297. c region or an FcRn-binding fragment thereof. In certain embodiments, the FcRn antagonist a plurality of FcRn antagonist molecules, wherein at least 50 of the plurality of FcRn antagonist molecules % (optionally at least 60, 70, 80, 90, 95, or 99%) of the nucleotides are bisecting at EU position 297. A variant Fc region or an FcRn-binding fragment thereof comprising an N-linked glycan having a binding GlcNac. include.

[0017] In certain embodiments, the variant Fc region is linked to a half-life extender. In certain embodiments, the half-life extender is polyethylene glycol or Human serum albumin.

[0018] In certain embodiments, the Fc-containing agent is an antibody or an immunoadhesin. In certain embodiments, the Fc-containing agent is a therapeutic or diagnostic agent. The Fc-containing agent is an imaging agent. In certain embodiments, the Fc-containing agent is an antibody drug conjugate. In certain embodiments, the Fc-containing agent is a pathogenic antibody. In an embodiment, the Fc-containing agent is an autoantibody.

[0019] In certain embodiments, the subject has an antibody-mediated disease or disorder, wherein the subject The administration of the FcRn antagonist ameliorates the disease or disorder. wherein the disease or disorder is treated with intravenous immunoglobulin (IVIG), plasma exchange therapy, and / or immunotherapy. In certain embodiments, the antibody-mediated disease or disorder is treatable using adsorption techniques. is an autoimmune disease. In certain embodiments, the autoimmune disease is allogeneic pancreatic cancer. Islet graft rejection, alopecia areata, ankylosing spondylitis, antiphospholipid syndrome, autoimmune amyloidosis Son's disease, Alzheimer's disease, antineutrophil cytoplasmic autoantibodies (ANCA), autoimmune diseases of the adrenal gland, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune myocarditis, autoimmune neutropenia, Autoimmune oophoritis and orchitis, autoimmune thrombocytopenia, autoimmune urticaria, Bechet's disease rheumatoid arthritis, bullous pemphigoid, cardiomyopathy, Castleman syndrome, celiac disease c spruce-dermatitis), chronic fatigue immune deficiency syndrome, chronic inflammatory demyelinating polyneuropathy (CIDP) ), Churg-Strauss syndrome, cicatricial pemphigoid, Crest syndrome, cold agglutinin disease, Crohn's disease, dermatomyositis, dilated cardiomyopathy, discoid lupus erythematosus , epidermolysis bullosa acquisita, essential mixed cryoglobulinemia, factor VIII deficiency, fibromyalgia Diseases - fibromyositis, glomerulonephritis, Graves' disease, Guillain-Barre, Goodpasture's syndrome, Graft-versus-host disease (GVHD), Hashimoto's thyroiditis, hemophilia A, idiopathic membranous neuropathy, idiopathic Pulmonary fibrosis, idiopathic thrombocytopenic purpura (ITP), IgA neuropathy, IgM polyneuropathy paci, immune-mediated thrombocytopenia, juvenile arthritis, Kawasaki disease, lichen plantus , lichen sclerosus, lupus erthematosis, Meniere's disease, mixed connective tissue fibrosis, mucous membrane pemphigoid, multiple sclerosis, type 1 diabetes, multifocal motor neuropathy (MMN), severe Myasthenia gravis, paraneoplastic bullous pemphigoid, gestational pemphigoid, pemphigus vulgaris, pemphigus foliaceus, Pernicious anemia, polyarteritis nodosa, polychrondritis, polyglandular syndrome, rheumatoid arthritis Polymyalgia machinosum, polymyositis and dermatomyositis, primary agammaglobulinemia agammaglobinulinemia), primary biliary cirrhosis, psoriasis, psoriatic arthritis, recurrent multiple cartilage ulcers inflammation, Raynaud's phenomenon, Reiter's syndrome, rheumatoid arthritis, Lucoidosis, scleroderma, Sjögren's syndrome, solid organ transplantation Rejection, stiff man syndrome, systemic lupus erythematosus, Takayasu's arteritis, toxic epidermal necrosis Ten-year-old Nephropathy (TEN), Stevens-Johnson Syndrome (SJS), Temporal Arteritis / Giant Cell Arteritis (temporal arteristis / giant cell arteritis), thrombotic thrombocytopenic purpura, large ulcerative Enteritis, uveitis, dermatitis herpetiformis vasculitis , antineutrophil cytoplasmic antibody-associated vasculitis, vitiligo, and Wegener's granulomatosis. can be.

[0020] In certain embodiments, the autoimmune disease is an autoimmune channelopathy. In embodiments, the channelopathy is autoimmune limbic encephalitis, epilepsy, neuromyelitis optica, Lambert-Eaton myasthenic syndrome, myasthenia gravis, anti-N-methyl-D-aspartic acid (N MDA) receptor encephalitis, anti-α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor encephalitis, Morvan syndrome, neuromyotonia, streptococcus Pediatric autoimmune neuropsychiatric disorders associated with bacterial infection (PANDAS) and glycine receptor antibody-related In certain embodiments, the antibody-mediated disorder is selected from the group consisting of hypersensitivity disorders, hyperlipidemia, and hypersensitivity disorders. He has globulinemia.

[0021] In certain embodiments, the FcRn antagonist is administered simultaneously or sequentially with an additional therapeutic agent. In certain embodiments, the additional therapeutic agent is an anti-inflammatory agent. In certain embodiments, the additional therapeutic agent is a leukoreducing agent. In certain embodiments, the leukocyte depleting agent is a B cell depleting agent. The drug is an antibody. In certain embodiments, the B cell depleting drug is an antibody that binds to CD10, CD19, CD20, CD2 1, CD22, CD23, CD24, CD37, CD53, CD70, CD72, CD74, CD75, CD77, CD79a, CD79b, CD8 It is an antibody that specifically binds to CD80, CD81, CD82, CD83, CD84, CD85, or CD86. In embodiments, the additional therapeutic agent is rituximab, daclizumab, basiliximab, mutagenesis agent, or vasopressin. Muronomab-CD3, infliximab, adalimumab, omalizumab, efavirenz rituximab, natalizumab, tocilizumab, eculizumab, golimumab, canakinumab, stekinumab, belimumab, or a combination thereof.

[0022] In certain embodiments, the subject is a human or a cynomolgus monkey. [Brief explanation of the drawings]

[0023] BRIEF DESCRIPTION OF THE DRAWINGS [Figure 1] FIG. 1 shows the results of an experiment to determine the effect of Fc-Abdeg and HEL-Abdeg on serum levels of a tracer antibody (FR70-hIgG1) in cynomolgous monkeys. [Figure 2] FIG. 2 shows the results of an experiment to determine the effect of Fc-Abdeg and HEL-Abdeg on serum levels of total IgG in cynomolgus monkeys. [Figure 3] FIG. 3 shows the results of experiments to determine the effects of Fc-Abdeg and HEL-Abdeg on albumin levels in cynomolgus monkeys. [Figure 4] FIG. 4 shows the results of an experiment to determine the effect of Fc-Abdeg and IVIG on serum levels of a tracer antibody (FR70-hIgG1) in cynomolgus monkeys. [Figure 5] FIG. 5 shows the results of an ELISA assay comparing the affinity of Fc-Abdeg, Fc-Abdeg-POT, and Fc-Abdeg-S239D / I332E for human CD16a. [Figure 6] FIG. 6 shows the results of an ELISA assay comparing the affinity of Fc-Abdeg, Fc-Abdeg-POT, and Fc-Abdeg-S239D / I332E for mouse CD16-2. [Figure 7] FIG. 7 shows the results of experiments to determine the effects of Fc-Abdeg, Abdeg-POT, and Fc-AbdegS239D / I332E on anti-CD20-induced ADCC signals using Promega's Raji-based ADCC reporter bioassay. [Figure 8] FIG. 8 shows the results of experiments to determine the effect of Fc-Abdeg and Abdeg-POT on anti-CD70-induced lysis of CD70+ U266 cells in vitro. [Figure 9] FIG. 9 shows the results of experiments to determine the effects of Fc-Abdeg, Fc-Abdeg-POT, Fc-Abdeg-S239D / I332E, and IVIG on platelet levels in an acute mouse model of immune thrombocytopenia. [Figure 10] FIG. 10 shows the results of an exemplary gel filtration purification of Fc-Abdeg. [Figure 11] FIG. 11 shows the results of a dose-escalation study measuring the effect of various single doses of Fc-Abdeg on serum levels of a tracer antibody (FR70-hIgG1) in cynomolgus monkeys. [Figure 12]FIG. 12 shows the results of a dose-escalation study measuring the effect of various single doses of Fc-Abdeg on serum levels of a tracer antibody (FR70-hIgG1) in cynomolgus monkeys. [Figure 13] FIG. 13 shows the results of an experiment to determine the effect of 200 mg / kg Fc-Abdeg on levels of cIgA in cynomolgus monkeys. [Figure 14] FIG. 14 shows the results of an experiment to determine the effect of 200 mg / kg Fc-Abdeg on levels of cIgM in cynomolgus monkeys. [Figure 15] FIG. 15 shows the pharmacokinetic profiles of various single doses of Fc-Abdeg in cynomolgus monkeys. [Figure 16] FIG. 16 shows the results of an experiment to determine the effect of repeated multiple doses of 20 mg / kg Fc-Abdeg on levels of cIgG in cynomolgus monkeys. [Figure 17] FIG. 17 shows the pharmacokinetic profile of Fc-Abdeg in cynomolgus monkeys given repeated multiple doses of 20 mg / kg Fc-Abdeg. [Figure 18] FIG. 18 shows the results of a dose-escalation study measuring the effect of various single doses of Fc-Abdeg on serum levels of endogenous IgG in cynomolgus monkeys. [Figure 19] FIG. 19 shows the pharmacokinetic profiles of various single doses of Fc-Abdeg in cynomolgus monkeys. [Figure 20] FIG. 20 shows the results of a repeat-dose study measuring the effect of various repeat doses of Fc-Abdeg on serum levels of endogenous IgG in cynomolgus monkeys. [Figure 21] FIG. 21 shows the pharmacokinetic profiles of various repeat doses of Fc-Abdeg in cynomolgus monkeys. [Figure 22] FIG. 22 shows the results of a sequential dosing study measuring the effect of various doses of Fc-Abdeg on serum levels of endogenous IgG in cynomolgus monkeys. [Figure 23] Figure 23 shows the results of a sequential dosing study measuring the effect of intravenous loading of a 20 mg / kg dose of Fc-Abdeg followed 24 hours later by daily subcutaneous administration of 3 mg / kg of Fc-Abdeg for 28 days, followed by a 32-day treatment-free period in cynomolgus monkey subjects. DETAILED DESCRIPTION OF THE INVENTION

[0024] (Detailed explanation) The present disclosure provides methods for measuring serum levels of Fc-containing agents (e.g., antibodies and immunoadhesins) in a subject. These methods generally involve administering to the subject a native Fc region. specifically binds to FcRn with increased affinity and reduced pH dependence compared to the The disclosed methods include administering an effective amount of an isolated FcRn antagonist to a subject. They are particularly useful in treating mediated disorders (eg, autoimmune diseases).

[0025] (I. Definition) Unless otherwise defined herein, scientific and technical terms used in connection with the present invention are The meaning and scope of the terms shall be as commonly understood by those skilled in the art. However, in the unlikely event that there is any hidden ambiguity, the definitions provided in this specification Furthermore, unless the context requires otherwise, Unless otherwise specified, singular terms shall include pluralities and plural terms shall include the singular. Generally, the methods described herein for cell and tissue culture, molecular biology, immunology, microbiology, genetics, Used in genetics, protein chemistry, nucleic acid chemistry, and hybridization. The terminology and techniques used are based on terminology and techniques well known and commonly used in the art. It's technology.

[0026] In order that the present invention may be more readily understood, certain terms are first defined.

[0027] As used herein, the term "FcRn antagonist" refers to an FcRn antagonist that specifically binds to FcRn via the Fc region. Fc regions (e.g., those disclosed herein) that bind to and inhibit the binding of immunoglobulins to FcRn. The term "antibody" refers to any agent containing a variant Fc region (provided the agent is not a full-length IgG antibody). .

[0028] As used herein, the term "Fc region" refers to the region formed by the Fc domains of the two heavy chains. The native Fc region is a homodimer. .

[0029] As used herein, the term "variant Fc region" refers to a region that has one or more Fc domains that differ in structure from a native Fc region. The term "Fc region" refers to an Fc region having the above alterations. The alterations include amino acid substitutions, additions, and / or deletions, This may include the attachment of additional moieties and / or alteration of the native glycan. The term encompasses heterodimeric Fc regions in which each of the constituent Fc domains is different. Examples of such heterodimeric Fc regions include, but are not limited to, "Knob and hole" as described in US 8216805, which is incorporated herein by reference. The term also includes, for example, Fc regions produced using techniques as described herein in their entirety. As described in US20090252729A1 and US20110081345A1, each of which is incorporated by reference. As shown in Figure 1, the component Fc domains are linked together by a linker moiety. The Fc region is also included.

[0030] As used herein, the term "Fc domain" refers to the hinge region immediately upstream of the papain cleavage site. It refers to the portion of a single immunoglobulin heavy chain that begins in the C-terminus region and ends at the C-terminus of the antibody. Thus, a complete Fc domain includes at least a portion of the hinge domain (e.g., the upper, middle, and and / or the lower hinge region), a CH2 domain, and a CH3 domain.

[0031] As used herein, the term "FcRn-binding fragment" refers to a portion of an Fc region sufficient to confer FcRn binding. Refers to the part.

[0032] The term "EU position" as used herein refers to the position of the EU site as defined by Edelman, GM et al., Proc. Natl. Acad. USA, 63, 78-85 (1969) and "Sequences of Proteins of Interest in Immunology" "Steins of Immunological Interest," U.S. Dept. Health and Human Services, 5th ed. Amino acids in the EU numbering convention for the Fc region as described in Kabat et al., 1991 Refers to the acid position.

[0033] As used herein, the term "CH1 domain" refers to the immunoglobulin domain extending from about EU position 118-215. The CH1 domain refers to the first (most amino-terminal) constant region domain of a globulin heavy chain. is adjacent to the VH domain and amino terminal to the hinge region of the immunoglobulin heavy chain molecule, It does not form part of the immunoglobulin heavy chain Fc region.

[0034] As used herein, the term "hinge region" refers to the region of the heavy chain that connects the CH1 domain with the CH2 domain. This hinge region contains approximately 25 residues and is flexible, allowing the two The hinge region separates the three distinct domains, allowing the N-terminal antigen-binding region of each to move independently. : upper, middle, and lower hinge domains (Roux et al., J. Immunol. 161: 4083 (1998) The FcRn antagonists of the present disclosure may be divided into all or part of the hinge. It can contain regions.

[0035] As used herein, the term "CH2 domain" refers to the heavy chain immune domain extending from about EU position 231-340. Refers to the part of the immunoglobulin molecule.

[0036] As used herein, the term "CH3 domain" refers to the region consisting of approximately 110 residues from the N-terminus of the CH2 domain. a portion of a heavy chain immunoglobulin molecule extending from, for example, about positions 341-446 (EU numbering system) include.

[0037] As used herein, the term "FcRn" refers to the neonatal Fc receptor. An exemplary FcRn molecule is It contains human FcRn encoded by the FCGRT gene described in RefSeq NM_004107.

[0038] As used herein, the term "CD16" refers to a protein required for antibody-dependent cell-mediated cytotoxicity (ADCC). An exemplary CD16 molecule is the human FcγRIII Fc receptor described in RefSeq NM_000569. Contains CD16a.

[0039] As used herein, the term "free cysteine" refers to a cysteine ​​that is present in a mature FcRn antagonist. Native or engineered cysteine ​​amino acid residues present in a qualitatively reduced form Refers to...

[0040] As used herein, the term "antibody" refers to a polypeptide chain consisting of four disulfide-linked polypeptide chains. An immunoglobulin molecule containing two heavy (H) chains and two light (L) chains interconnected by bonds Each heavy chain comprises a heavy chain variable region (VH and The heavy chain constant region contains three domains, CH1, CH2, and CH3. Each light chain comprises a light chain variable region (abbreviated as VL) and a light chain constant region. The chain constant region contains one domain (CL). The VH and VL regions are divided into framework regions (FR). The complementarity-determining regions (CDRs) are interspersed with more conserved regions called These can be further subdivided into hyperdenatured regions.

[0041] As used herein, the term "N-linked glycan" refers to a glycan present in the CH2 domain of the Fc region. sequons (i.e., Asn-X-Ser or Asn-X-Thr sequences, where X is any amino acid except proline) It refers to an N-linked glycan attached to the nitrogen (N) in the side chain of asparagine in Such N-glycans can be found, for example, in the literature, which is incorporated herein by reference in its entirety. Drickamer K and Taylor ME (2006) Introduction to Glycobiology. This is fully described in the second edition of "The Biology of the Earth."

[0042] The term "afcosylated" as used herein is intended to mean a compound that is incorporated herein by reference in its entirety. lacking a core fucose molecule, as described in US8067232, incorporated herein by reference. It refers to N-linked glycans.

[0043] As used herein, the term "bisecting GlcNac" refers to the same as defined herein in its entirety. a core mannose molecule, as described in U.S. Pat. No. 8,021,856, incorporated by reference in its entirety. refers to an N-linked glycan having an N-acetylglucosamine (GlcNAc) molecule attached to it.

[0044] As used herein, the term "antibody-mediated disorder" refers to a disorder caused by the presence of antibodies in a subject. This refers to any disease or disorder that causes or worsens a person's condition.

[0045] As used herein, the term "Fc-containing agent" is any molecule that contains an Fc region.

[0046] As used herein, the term "leukocytapheresis agent" refers to an agent that, upon administration, reduces the number of white blood cells in a subject. Refers to drugs that decrease blood cholesterol levels.

[0047] As used herein, the term "B cell depleting agent" refers to an agent that, upon administration, reduces the number of B cells in a subject. This refers to a drug that lowers blood pressure.

[0048] As used herein, the term "T cell depleting agent" refers to an agent that, upon administration, reduces the number of T cells in a subject. This refers to a drug that lowers blood pressure.

[0049] As used herein, the term "autoimmune channelopathy" refers to a disorder characterized by the loss of a subunit of an ion channel. This refers to a disease caused by autoantibodies against the antibody or molecules that regulate the channel.

[0050] As used herein, the terms "treat," "treating," and "treatment" refer to The method of "treatment" refers to the therapeutic or prophylactic measures described in the document. Subjects with a disease or disorder (e.g., inflammation and cancer) or those suffering from such a disease or disorder a subject predisposed to have one or more of the disease or disorder, or a recurrence of the disease or disorder; to prevent, cure, delay, reduce the severity of, or ameliorate the symptoms of, or In order to prolong the survival of a subject beyond that expected without such treatment, The term "subject" as used herein refers to a subject that is administered an antibody or antigen-binding fragment thereof. " includes any human or non-human animal.

[0051] As used herein, the term "immunoadhesin" refers to a binding protein ( refers to antibody-like molecules that contain functional domains of a specific molecule (e.g., a receptor, a ligand, or a cell adhesion molecule) .

[0052] II. Methods of Reducing Serum Levels of Fc-Containing Agents In one aspect, the present disclosure provides a method for administering an Fc-containing agent (e.g., an antibody or immunoadhesin) to a subject. a method for reducing serum levels of a phosphodiesterase (Fc) domain, comprising administering to said subject a phosphodiesterase (Fc) domain relative to a native Fc region; an effective amount of an isolated antibody that specifically binds to FcRn with increased affinity and reduced pH dependence; administration of an FcRn antagonist (e.g., an FcRn antagonist disclosed herein) The method further comprises:

[0053] As provided herein, administration of an FcRn antagonist to the subject at a dose of about 0.2 to about 200 mg / kg is Thus, in certain embodiments, administration of the FcRn antigen The antagonist is administered to the subject at a dose of about 0.2 to about 200 mg / kg (e.g., 0.2 to 200 mg / kg). In certain embodiments, the FcRn antagonist is administered in an amount of about 0.2, 2, 20, 70, or 200 mg. The subject is administered a dose of 0.2, 2, 20, 70, or 200 mg / kg (e.g., 0.2, 2, 20, 70, or 200 mg / kg). In an embodiment, the FcRn antagonist is administered at a dose of about 20 mg / kg (e.g., 20 mg / kg). is administered to the subject.

[0054] As shown herein, multiple repeat administration regimes unexpectedly provide better efficacy than single administrations. Thus, in certain embodiments, the FcRn antagonist has a therapeutic effect of at least 20 days. In certain embodiments, the FcRn antagonist is administered to the subject twice. The compound is administered to the subject once every 4, 5, 6, 7, 8, 9, or 10 days. The FcRn antagonist was administered on days 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, In certain embodiments, the FcRn antigen is administered to the subject 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 times. In certain embodiments, the FcRn antagonist is administered to the subject once every four days. Antagonist was administered every 4 days for 13 days (i.e., days 1, 5, 9, and 13). In certain embodiments, 20 mg / kg of an FcRn antagonist is administered to the subject. The subject will be administered a dose every four days (ie, on days 1, 5, 9, and 13) over a period of one year.

[0055] The FcRn antagonist can be administered to the subject by any means. These include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, and epidural. The compositions can be administered, for example, by infusion or bolus injection. In certain embodiments, the FcRn antagonist is administered by intravenous infusion.

[0056] The methods disclosed herein can reduce serum levels of any Fc-containing agent. In certain embodiments, the Fc-containing agent is an antibody or an immunoadhesin. In certain embodiments, the Fc-containing agent is a therapeutic or diagnostic agent. The Fc-containing agent is an imaging agent. In certain embodiments, the Fc-containing agent is an antibody drug conjugate. In certain embodiments, the Fc-containing agent is a conjugate that inhibits pathogenic antibodies, e.g., autoantibodies. In certain embodiments, the subject has an antibody-mediated disease or disorder. In one embodiment, the antibody-mediated disease or disorder is associated with autoantibodies.

[0057] The reduction in serum levels of Fc-containing agents (e.g., antibodies and immunoadhesins) is due to antibody-mediated It is particularly applicable to the treatment of disorders (e.g., autoimmune diseases). The present disclosure relates to methods of treating a subject having an antibody-mediated disorder (e.g., an autoimmune disease). administering to the subject an effective amount of an FcRn antagonist composition disclosed herein. The method further comprises:

[0058] Any antibody-mediated disorder is treatable using the methods disclosed herein. In certain embodiments, the antibody-mediated disorder is a disorder amenable to treatment with IVIG. In one embodiment, the antibody-mediated disorder is an autoimmune disease. Epidemic diseases include allogeneic islet graft rejection, alopecia areata, ankylosing spondylitis, and antiphospholipid Somatic syndrome, autoimmune Addison's disease, Alzheimer's disease, antineutrophil cytoplasmic autoantibodies (ANCA) , autoimmune diseases of the adrenal gland, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune myocarditis, Autoimmune neutropenia, autoimmune oophoritis and orchitis, autoimmune thrombocytopenia, autoimmune Autoimmune urticaria, Behçet's disease, bullous pemphigoid, cardiomyopathy, Castleman syndrome, Riac disease - dermatitis, chronic fatigue immunodeficiency syndrome, chronic inflammatory demyelinating polyneuritis (CIDP), Churg-Strauss syndrome, cicatricial pemphigoid, CREST syndrome, cold agglutinin disease, claws Disease, dermatomyositis, discoid lupus erythematosus, essential mixed cryoglobulinemia, factor VIII Factor deficiency, fibromyalgia-fibromyositis, glomerulonephritis, Graves' disease, Guillain-Barré, Goodman disease Depasture's syndrome, graft-versus-host disease (GVHD), Hashimoto's thyroiditis, hemophilia A, idiopathic pulmonary fibrosis Idiopathic thrombocytopenic purpura (ITP), IgA neuropathy, IgM polyneuropathy , immune-mediated thrombocytopenia, juvenile arthritis, Kawasaki disease, lichen planus, lupus erythematosus, meningitis Yale disease, mixed connective tissue disease, multiple sclerosis, type 1 diabetes, multifocal motor neuropathy (M MN), myasthenia gravis, paraneoplastic bullous pemphigoid, pemphigus vulgaris, pemphigus foliaceus, pernicious anemia , polyarteritis nodosa, polychondritis, polyglandular syndrome, polymyalgia rheumatica, polymyositis and dermatomyositis, primary agammaglobulinemia, primary biliary cirrhosis, psoriasis, psoriatic joints inflammation, Raynaud's phenomenon, Reiter's syndrome, rheumatoid arthritis, sarcoidosis, scleroderma, Sjogren's syndrome, solid organ transplant rejection, stiff man syndrome, systemic lupus erythematosus Death, Takayasu's arteritis, toxic epidermal necrolysis (TEN), Stevens-Johnson syndrome (SJ) S), temporal arteritis / giant cell arteritis, thrombotic thrombocytopenic purpura, ulcerative colitis, uveal ulcer dermatitis, dermatitis herpetiformis vasculitis, antineutrophil cytoplasmic antibody-associated vasculitis, vitiligo, and Wegener's disease If you have blastoma.

[0059] In certain embodiments, the autoimmune disease is an autoimmune channelopathy. Notable channelopathies include neuromyelitis optica, Lambert-Eaton myasthenic syndrome, and myasthenia gravis. , anti-N-methyl-D-aspartate (NMDA) receptor encephalitis, anti-α-amino-3-hydroxy-5-methyl Amphi-4-isoxazolepropionic acid (AMPA) receptor encephalitis, Morvan's syndrome, and glycine There is a receptor antibody-related disorder.

[0060] The disclosed methods are directed to treating antibody-mediated disorders characterized by excessive production of serum immunoglobulins. Thus, in certain embodiments, the FcRn antagonist The composition is used to treat hypergammaglobulinemia.

[0061] The methods of the present disclosure can also be used in combination with one or more additional therapeutic agents. In certain embodiments, the additional therapeutic agent is an anti-inflammatory agent. In certain embodiments, the compositions can be used in combination with the compositions disclosed in the literature. Treatment options include rituximab, daclizumab, basiliximab, muronomab-cd3, and infliximab. Cimab, adalimumab, omalizumab, efalizumab, natalizumab, tocilizumab, eculizumab, golimumab, canakinumab, ustekinumab, or belimumab. In certain embodiments, the additional therapeutic agent is a leukodepleting agent (e.g., a B cell or T cell depleting agent). Any leukocyte-depleting agent can be combined with the FcRn antagonist compositions disclosed herein. In certain embodiments, the leukocytapheresis agent is a B cell leukocyte depletion agent. In certain embodiments, the leukocyte depleting agent is a leukocyte depleting agent directed against a cell surface marker. Suitable cell surface markers include, but are not limited to, CD10, CD19, CD20, and CD21. , CD22, CD23, CD24, CD37, CD53, CD70, CD72, CD74, CD75, CD77, CD79a, CD79b, CD80 , CD81, CD82, CD83, CD84, CD85, or CD86. The therapeutic agent(s) may be administered simultaneously or sequentially via the same or different route(s). , can be administered to the subject.

[0062] The disclosed methods also provide rapid reduction of serum levels of Fc-containing agents in a subject. Such rapid clearance reduces the subject's exposure to the drug. Therefore, the Fc-containing agent (e.g., an antibody drug conjugate or an agent that is immunogenic) may exhibit toxicity. Rapid clearance also allows Fc-containing agents to facilitate imaging. This is advantageous when the agent is a contrast agent that requires low serum levels for this purpose. In certain embodiments, the FcRn antagonist composition is administered to a subject who has received the Fc-containing agent. It is used to reduce the serum level of an Fc-containing agent (e.g., an imaging agent) in a subject. Serum levels of any Fc-containing agent (e.g., a therapeutic or diagnostic agent) can be measured using FcRn as disclosed herein. The Fc-containing agent can be reduced using an antagonist composition. Non-limiting examples of Fc-containing agents include: Imaging agents (e.g., labeled antibodies), antibody drug conjugates, or immunogenic agents (e.g., non- The FcRn antagonist is a human antibody or immunoadhesin. The Fc-containing agent may be administered simultaneously or sequentially (eg, before or after the Fc-containing agent).

[0063] Furthermore, in diseases or conditions that require the administration of a therapeutic agent, the subject often has a high tolerance to the agent. The antibody produces antibodies against the drug (e.g., anti-drug antibodies), which in turn inhibit the therapeutic agent's intended treatment. prevent it from being available for therapeutic purposes or cause adverse reactions in the subject. Thus, the methods disclosed herein involve detecting antibodies ( For example, it can also be used to remove anti-drug antibodies.

[0064] The methods disclosed herein also provide therapeutic targeting by reducing levels of IgG. It can be used in combination with therapeutic proteins to enhance the benefits of proteins. where IgG antibodies contribute to reduced bioavailability of therapeutic proteins. In one embodiment, the present disclosure provides a method for treating a patient with a disorder resulting from an immune response to a blood clotting factor. A method of treating a subject, comprising administering to the subject a therapeutically effective amount of an FcRn antagonist disclosed herein. The method further comprises administering a coagulation factor to a patient having a blood coagulation disorder. Not specified, but fibrinogen, prothrombin, factor V, factor VII, and factor VIII , Factor IX, Factor X, Factor XI, Factor XII, Factor XIII, or von Willebrand This method is useful for the treatment of blood clotting factors, for example, in patients with hemophilia A or B. It can be used to regulate, treat, or prevent immune responses to the offspring. In one embodiment, the method is for treating, for example, a patient suffering from pure red cell aplasia (PRCA). It can be used to regulate or treat the immune response to erythropoietin.

[0065] In pregnant women, FcRn contributes to the transport of maternal antibodies across the placenta to the fetus. When a pregnant woman is administered an Fc-containing agent (e.g., a therapeutic antibody), the agent is delivered via the placenta to the Fc As a result of Rn-mediated transport, the Fc-containing agent may come into contact with the fetus. It would be beneficial to block FcRn function to avoid any potential adverse effects on immune cells. Therefore, the present disclosure provides a method for preventing the delivery of Fc-containing agents (e.g., therapeutic antibodies) to a fetus in a pregnant woman. a method for preventing placental transfer of a compound comprising administering to said woman an FcRn antagonist as disclosed herein; administering the composition simultaneously or sequentially (before or after) the Fc-containing agent. A method is provided.

[0066] The methods disclosed herein also include, but are not limited to, treatments for asthma, ulcerative colitis, and the like. and inflammatory disorders including inflammatory bowel syndrome, allergic rhinitis / sinusitis, skin allergies ( Urticaria / hives, angioedema, atopic dermatitis, food allergies Allergies including drug allergies, insect allergies, mastocytosis, osteoarthritis, It can be used to treat arthritis, including eumatoid arthritis and spondyloarthropathies.

[0067] Successful implementation of gene therapy for the treatment of a disease or condition depends on the gene encoding the transgene. The vector used to deliver the therapeutic protein and possibly the transgene This can be prevented by the production of antibodies specific to the FcRn antigen. Administration of the antagonist composition in combination with gene therapy reduces IgG levels. These methods can enhance the benefits of the encoded therapeutic protein. IgG antibodies are expressed as a reduced biological effect of a gene therapy vector or an encoded therapeutic protein. The gene therapy vector is particularly useful in situations where availability is a factor. They can be viral vectors such as adenovirus and adeno-associated virus. Diseases that can be treated using gene therapy include, but are not limited to, cystic leukemia, fibrosis, hemophilia, PRCA, muscular dystrophy, or lysosomal storage diseases, such as Gaucher disease and and Fabry disease.

[0068] Any subject can be treated using the methods disclosed herein. In embodiments, the subject is a human or a cynomolgus monkey.

[0069] III. FcRn Antagonists The methods disclosed herein generally involve administering to a subject an effective amount of an isolated FcRn antagonist. wherein the FcRn antagonist induces increased FcRn activity compared to a native Fc region. They specifically bind to FcRn with increased affinity and reduced pH dependence. FcRn antagonists have increased affinity and decreased pH dependence compared to native Fc regions. The variant Fc region or FcRn-binding fragment thereof specifically binds to FcRn with a specificity dependent on the FcRn-binding domain. The FcRn antagonist interacts with Fc-containing agents (e.g., antibodies and immunoadhesins) in vivo. inhibits Rn binding, thereby increasing the rate of degradation of Fc-containing agents and, at the same time, inhibits the degradation of these agents serum levels of

[0070] As provided herein, certain isolated variant Fc regions (e.g., those at EU positions 252 A variant Fc region containing amino acids Y, T, E, K, F, and Y at positions 254, 256, 433, 434, and 436. The variant Fc region is a more effective FcRn antagonist in vivo than a full-length antibody containing the same variant Fc region. Thus, in certain embodiments, the FcRn antagonist composition is a full-length FcRn antagonist. In certain embodiments, the FcRn antagonist composition is not an antibody. In certain embodiments, the FcRn antagonist composition does not comprise an antibody variable domain. However, in certain embodiments, the FcRn antagonist does not comprise a CH1 domain. The immunostimulatory composition may be linked to one or more additional binding domains or binding moieties, including antibody variable domains. The antibody may comprise a variant Fc region linked to the antibody.

[0071] Any Fc region may be altered to be included in the FcRn antagonist compositions disclosed herein. Variant Fc regions can be produced for use. Generally, the Fc region or its FcRn The binding fragments are derived from human immunoglobulins. However, the Fc region may be derived from, for example, camelids. species, rodents (e.g., mice, rats, rabbits, guinea pigs) or non-human primates ( derived from immunoglobulins of any other mammalian species, including, for example, chimpanzees, macaques, etc. It will be understood that the Fc region or a portion thereof may also be a fragment of IgM, IgG, IgD, IgA, or the like. and any immunoglobulin class, including IgE, and any class, including IgG1, IgG2, IgG3, and IgG4. In certain embodiments, the Fc region may be derived from any immunoglobulin isotype. is an IgG Fc region (e.g., a human IgG region). In certain embodiments, the Fc region is an IgG In certain embodiments, the Fc region is A suitable example of a chimeric Fc region is a chimeric Fc region that contains portions of two or more different Fc regions. The various F c-region gene sequences (e.g., human constant region gene sequences) are available in publicly deposited form It is understood that the scope of the present invention includes alleles, variants and mutations of the Fc region. Let's do it.

[0072] The Fc region can be further truncated or internally deleted to produce its minimal FcRn-binding fragment. The ability of the Fc region fragment to bind to FcRn can be determined by any art-recognized binding assay. The antibody titer can be determined using, for example, ELISA.

[0073] To enhance the manufacturability of the FcRn antagonists disclosed herein, the components It is preferred that the Fc region does not contain any cysteine ​​residues that do not form disulfide bonds. Accordingly, in certain embodiments, the Fc region does not comprise a free cysteine ​​residue.

[0074] FcRn binds with increased affinity and reduced pH dependence compared to the native Fc region. Any Fc variant or FcRn-binding fragment thereof that specifically binds to the FcRn-binding fragments disclosed herein. In certain embodiments, a variant can be used in an FcRn antagonist composition. The Fc region contains amino acid changes, substitutions, insertions and / or deletions that confer desired properties. In an embodiment, the variant Fc region or fragment comprises a nucleotide sequence at EU positions 252, 254, 256, 433, 440, 442, 443, 444, 445, 446, 447, 448, 449, 450, 451, 452, 453, 454, 455, 456, 457, 45 , 434, and 436 contain amino acids Y, T, E, K, F, and Y. Can be used in variant Fc regions Non-limiting examples of suitable amino acid sequences are listed in Table 1 herein. wherein the amino acid sequence of the Fc domain of the variant Fc region is set forth in SEQ ID NO: 1. In certain embodiments, the variant Fc region comprises the amino acid sequence of the Fc domain. The amino acid sequence consists of the amino acid sequence set forth in SEQ ID NO: 1, 2, or 3. wherein the FcRn antagonist comprises a variant Fc region, The amino acid sequence of the Fc domain of the present invention is the amino acid sequence set forth in SEQ ID NO: 1, 2, or 3. do.

[0075] Table 1. Non-limiting example amino acid sequences of variant Fc regions [Table 1]

[0076] In certain embodiments, the variant Fc region has an altered ( The variant Fc region may have one or more Fc Gamma receptors, e.g., FcγRI (CD64), FcγRIIA (CD32), FcγRIIB (CD32), FcγRIIIA (CD 16a), and altered (e.g., increased or decreased) binding to FcγRIIIB (CD16b). Any technique known in the art that alters affinity for additional Fc receptors may be used. Art-recognized methods are available. In certain embodiments, the variant Fc The amino acid sequence of the region is altered.

[0077] In certain embodiments, the variant Fc region is selected from the group consisting of: Numbered 234, 235, 236, 239, 240, 241, 243, 244, 245, 247, 252, 254, 25 6, 262, 263, 264, 265, 266, 267, 269, 296, 297, 298, 299, 313, 325, 326, 327, 32 329, 330, 332, 333, and 334. Optionally, the Fc region may contain additional and / or alternative amino acid residues known to those of skill in the art. positions can contain amino acid residues that do not occur naturally (e.g., the entire content of U.S. Patent Nos. 5,624,821; 6,277,375; 6,737,056, incorporated herein by reference. No.; PCT Patent Publication Nos. WO 01 / 58957; WO 02 / 06919; WO 04 / 016750; WO 04 / 02920 No. 7; WO 04 / 035752 and WO 05 / 040217).

[0078] In certain embodiments, the variant Fc region is selected from the group consisting of: Numbered 234D, 234E, 234N, 234Q, 234T, 234H, 234Y, 234I, 234V, 234F, 23 5A, 235D, 235R, 235W, 235P, 235S, 235N, 235Q, 235T, 235H, 235Y, 235I, 235V, 235F , 236E, 239D, 239E, 239N, 239Q, 239F, 239T, 239H, 239Y, 2401, 240A, 240T, 240M, 241W, 241 L, 241Y, 241E, 241R, 243W, 243L 243Y, 243R, 243Q, 244H, 245A, 247V, 24 7G, 252Y, 254T, 256E, 262I, 262A, 262T, 262E, 263I, 263A, 263T, 263M, 264L, 264I , 264W, 264T, 264R, 264F, 264M, 264Y, 264E, 265G, 265N, 265Q, 265Y, 265F, 265V, 265I, 265L, 265H, 265T, 266I, 266A, 266T, 266M, 267Q, 267L, 269H, 269Y, 269F, 26 9R, 296E, 296Q, 296D, 296N, 296S, 296T, 296L, 296I, 296H, 269G, 297S, 297D, 297E , 298H, 298I, 298T, 298F, 299I, 299L, 299A, 299S, 299V, 299H, 299F, 299E, 313F, 325Q, 325L, 325I, 325D, 325E, 325A, 325T, 325V, 325H, 327G, 327W, 327N, 327L, 32 8S, 328M, 328D, 328E, 328N, 328Q, 328F, 328I, 328V, 328T, 328H, 328A, 329F, 329H , 329Q, 330K, 330G, 330T, 330C, 330L, 330Y, 330V, 330I, 330F, 330R, 330H, 332D, Selected from the group consisting of 332S, 332W, 332F, 332E, 332N, 332Q, 332T, 332H, 332Y, and 332A Optionally, the Fc region comprises at least one non-naturally occurring amino acid residue. The present invention may include additional and / or alternative non-naturally occurring amino acid residues known to those skilled in the art (e.g., See, for example, U.S. Patent No. 5,624,821, the entire contents of which are incorporated herein by reference; Nos. 6,277,375; 6,737,056; PCT Patent Publication Nos. WO 01 / 58957; WO 02 / 06919; WO See WO 04 / 016750; WO 04 / 029207; WO 04 / 035752 and WO 05 / 040217 stomach).

[0079] Other known Fc variants that can be used in the FcRn antagonists disclosed herein is incorporated herein by reference in its entirety, without limitation. ie et al., 1997, Nat. Biotech. 15:637-40; Duncan et al., 1988, Nature 332:563- 564; Lund et al., 1991, J. Immunol., 147:2657-2662; Lund et al., 1992, Mol. Immun. munol., 29:53-59; Alegre et al., 1994, Transplantation 57:1537-1543; Hutchins et al. 1995, Proc Natl. Acad Sci USA, 92:11980-11984; Jefferis et al., 1995, I mmunol Lett., 44:111-117; Lund et al., 1995, Faseb J., 9:115-119; Jefferis et al. Lund et al., 1996, J. Immunol., 157:4963 -4969; Armour et al., 1999, Eur J Immunol 29:2613-2624; Idusogie et al., 2000, J. Immunol., 164:4178-4184; Reddy et al., 2000, J. Immunol., 164:1925-1933; Xu et al., 2000, Cell Immunol., 200:16-26; Idusogie et al., 2001, J. Immunol., 166:2571-2575; Shields et al., 2001, J Biol. Chem., 276:6591-6604; Jefferis et al. 2002, Immunol Lett., 82:57-65; Presta et al., 2002, Biochem Soc Trans., 3 0:487-490); U.S. Patent Nos. 5,624,821; 5,885,573; 5,677,425; 6,165,745; No. 6,277,375; No. 5,869,046; No. 6,121,022; No. 5,624,821; No. 5,648,260; No. 6,5 Nos. 28,624; 6,194,551; 6,737,056; 6,821,505; 6,277,375; U.S. Patent Publication Patent Publication No. 2004 / 0002587 and PCT Publication Nos. WO 94 / 29351; WO 99 / 58572; WO 00 / 42072; The methods disclosed in WO 02 / 060919; WO 04 / 029207; WO 04 / 099249; WO 04 / 063351 Fc variants.

[0080] In certain embodiments, the variant Fc region is a heterodimer, wherein the constituent elements The constituent Fc domains are different from each other. Methods for producing Fc heterodimers are known in the art. are known in the art (see, for example, US 8216805, which is incorporated herein by reference in its entirety). In certain embodiments, the variant Fc region is a single chain Fc region, where the component Fc domains are linked together by linker moieties. Methods for producing the same are known in the art (e.g., see refs. 1 and 2, each of which is incorporated herein in its entirety). See US20090252729A1 and US20110081345A1, which are incorporated by reference.

[0081] Pathogenic IgG antibodies observed in autoimmune diseases are thought to be the pathogenic triggers of these diseases. These may contribute to or contribute to disease progression and may contribute to the progression of disease through inappropriate activation of cellular Fc receptors. It is believed to be a vehicle for the formation of aggregated autoantibodies and / or autoantigens complexed with Autoantibodies (immune complexes) bind to activated Fc receptors and cause numerous autoimmune diseases. causing inflammatory bowel disease (which is caused in part by immunologically mediated inflammation directed against self-tissue) (See, e.g., Clarkson et al., each of which is incorporated by reference in its entirety. References: NEJM 314(9), 1236-1239 (2013); US20040010124A1; US20040047862A1; and US2004 (See, e.g., US Pat. No. 6,265,321A1). Thus, antibody-mediated disorders (e.g., autoimmune diseases) can be prevented. Treatment involves removing harmful autoantibodies and activating Fc receptors of these antibodies. Both by blocking immune complex interactions with receptors (e.g., Fcγ receptors such as CD16a) and by blocking immune complex interactions with receptors (e.g., Fcγ receptors such as CD16a). It would be more beneficial.

[0082] Thus, in certain embodiments, the variant Fc region of the FcRn antagonist is CD16a (e.g., human CD16a). This indicates that the FcRn antagonist However, immune complex-induced inflammatory responses and autoantibodies are targeted for clearance by FcRn inhibition. It is particularly advantageous in that it can antagonize CD16a (e.g., human CD16a Any art-recognized method for increasing affinity for the ATP can be utilized. In certain embodiments, the FcRn antagonist comprises an N-linked group (e.g., at EU position 297). In this case, the glycan structure is altered. This may increase the binding affinity of the FcRn antagonist to CD16a. Alterations of the N-linked glycans in the c region are well known in the art. For example, afucosylated N- N-glycans with linked glycans or bisecting GlcNac structures bind to CD16a. It has been shown that N-bonds exhibit increased affinity. The glycans are afucosylated. Afucosylation is an art-recognized This can be achieved by any method that is suitable for this purpose. For example, the FcRn antagonist can be a fucosylated FcRn antagonist. The transferase can be expressed in cells deficient in the The N-linked glycan at EU position 297 of the resident Fc region is not attached to the glycan (e.g., the entire (See US 8,067,232, the entire disclosure of which is incorporated herein by reference.) Specific Embodiments In the present invention, the N-linked glycan has a bisecting GlcNac structure. The GlcNac structure can be achieved using any art-recognized method. For example, FcRn antagonists inhibit β1-4-N-acetylglucosaminyltransferase III (G nTIII), resulting in the bisecting GlcNac is added to the N-linked glycan at EU position 297 of the variant Fc region (e.g., (See US 8021856, the entirety of which is incorporated herein by reference). Alternatively, the alteration of the N-linked glycan structure can also be achieved by in vitro enzymatic methods. It can be achieved.

[0083] In certain embodiments, the FcRn antagonist comprises multiple FcRn antagonist molecules. wherein at least 50% (optionally, at least 60, 70, 80, 90, 95, or 99%) contain an afucosylated N-linked glycan at EU position 297 A variant Fc region, or an FcRn-binding fragment thereof.

[0084] In certain embodiments, the FcRn antagonist comprises multiple FcRn antagonist molecules. wherein at least 50% (optionally, at least 60, 70, 80, 90, 95, or 99%) are N-linked glutamic acid with a bisecting GlcNac at EU position 297 The variant Fc region comprises a glycan, or an FcRn-binding fragment thereof.

[0085] In certain embodiments, the variant Fc region does not contain N-linked glycans. This can be achieved using any art-recognized method. For example, can be expressed in cells that are incapable of N-linked glycosylation. Alternatively, the amino acid sequence of the Fc variant may be modified (e.g., by mutation of the NXT sequon). The amino acid sequence of the amino acid sequence of the present invention can be altered (e.g., by the addition of an amino acid sequence of the present invention) to prevent or inhibit N-linked glycosylation. Fc variants can be synthesized in a cell-free system (eg, chemical synthesis).

[0086] In certain embodiments, the FcRn antagonist specifically binds to FcRn by a covalent bond. For example, the FcRn antagonist may be provided with a molecule (e.g., a molecule that binds or The FcRn antagonist molecule can be modified by covalently attaching a compound (e.g., an imaging moiety) to the FcRn antagonist molecule. For example, but not by way of limitation, the FcRn antagonist may be a glycosylated, acetylated, or PEGylation, phosphorylation, amidation, derivatization with known protective blocking groups, proteolytic cleavage The polypeptides can be modified, for example, by linkage to cellular ligands or other proteins.

[0087] In certain embodiments, the FcRn antagonist is linked to a half-life extender. As used herein, the term "half-life extender" refers to a variant Fc region of the present invention. When linked to the FcRn antagonists disclosed herein, the half-life of the FcRn antagonist is Any molecule that increases half-life (covalently or non-covalently) In certain embodiments, the FcRn antagonist may be linked to the FcRn antagonist. Half-life extenders are polyethylene glycol or human serum albumin. In an embodiment, the FcRn antagonist is a half-life extender present in the subject. a binding molecule that specifically binds to the antibody, such as serum albumin (e.g., human serum albumin) The antibody is linked to a blood-borne molecule or cell, such as IgG, red blood cells, etc.

[0088] IV. PHARMACEUTICAL COMPOSITIONS In certain embodiments, the method comprises administering an FcRn antagonist or FcRn antagonist disclosed herein. A pharmaceutical composition comprising an Rn antagonist composition and a pharmaceutically acceptable carrier or excipient is provided. Examples of suitable pharmaceutical carriers are listed in "Remington's Pharmaceutical Sciences" Examples of excipients include starch, glucose, , lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, stevia Sodium stearate, glycerol monostearate, talc, sodium chloride, dried starch Examples include kimmed milk, glycerol, propylene glycol, water, and ethanol. The composition can also contain pH buffering agents, and wetting or emulsifying agents.

[0089] The pharmaceutical compositions are formulated for parenteral administration (e.g., intravenous or intramuscular) by bolus injection. Injectable preparations can be formulated in unit dosage form, for example, ampoules with added preservatives. The compositions may be presented in the form of a suspension, solution, or oil. The compositions may take such forms as emulsions in aqueous or aqueous vehicles, and suspensions, It may contain formulatory agents such as stabilizers and / or dispersants. Alternatively, the active ingredient may be in the form of a powder for constitution with a suitable vehicle, e.g., pyrogen-free water. It can be in the form of.

[0090] In the disclosed methods, the FcRn antagonist is a compound described in U.S. Pat. No. 5,326,856. The peptide-chelator conjugate can be linked to a chelator such as The compound is then radiolabeled and used to prepare a compound for the diagnosis or treatment of diseases or conditions involving modulation of IgG levels. A contrast agent can be provided. [Example]

[0091] V. Working Examples The present invention is further illustrated by the following examples, which should not be construed as further limiting. The sequence listing, figures, and all references cited throughout this application The contents of all such patents and published patent applications are expressly incorporated herein by reference.

[0092] Example 1: Effect of Fc-Abdeg on serum IgG levels in cynomolgus monkeys The serum IgG levels of tracer antibodies against human anti-lysozyme IgG (HEL-Abdeg) and Human chromatid containing amino acids Y, T, E, K, F, and Y at EU positions 252, 254, 256, 433, 434, and 436 The effect of the IgG Fc region (Fc-Abdeg) (Fc-Abdeg; SEQ ID NO: 2) was determined in cynomolgus monkeys Specifically, cynomolgus monkeys were injected with 1 mg / kg of anti-mouse CD70 hIgG1 tracer antibody (FR70-hI gG1; Oshima et al., Int Immunol 10(4): 517-26 (1998)) by intravenous bolus injection. Five minutes later, animals were given either 7 mg / kg Fc-Abdeg, 20 mg / kg HEL-Abdeg, or PBS. The infusion was administered within 1 hour and the animals were given a volume of 10 ml / kg. Blood samples (3 x 150 μl) were taken 5 minutes before dosing ("pre-dose") and at the end of the infusion. Samples were collected 5 minutes, 2 hours, 6 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, and 120 hours after the end of the experiment. Tracer levels were determined by performing mCD70 binding ELISA, and data were collected from the end-of-dose tracer. The total cynomolgus IgG levels were also determined. These experimental results show that Fc-Abdeg acts as a more efficient tracer antibody than an equivalent amount of HEL-Abdeg. This shows that the

[0093] In addition to its important role in the IgG recycling pathway, FcRn is also involved in albumin homeostasis. (Chaudhury et al., J Exp Med. 197(3):315-22 (2003)). FcRn regulates IgG-Fc and arginine-associated proteins. The binding of phospholipids to phospholipids can occur simultaneously (Andersen et al., Natl. Clin. mmun. 3:610 (2012)). Conceptually, blocking IgG recycling using Abdeg-modified molecules is Therefore, albumin-FcRn interaction should not be interfered with. This hypothesis is supported by in vitro studies in mice. This was confirmed in a study in which the authors demonstrated that hIgG1 molecules with Abdeg bind to albumin. showed no effect on the levels of erythrocytes (Patel et al., J Immunol 187(2): 1015-22 (20 11)). In the experiment described above, albumin was measured on days -3, 3, and 17 after the end of the infusion. As in the mouse study, albumin levels were also determined after treatment with Fc-Abdeg or HEL-Abdeg. No significant changes in min levels were observed (see Figure 3).

[0094] In subsequent experiments, the antibody removal efficacy of Fc-Abdeg was compared with that of IVIG. Two days before administration of Fc-Abdeg or 2 g / kg IVIG, 1 mg / kg tracer antibody (FR70-hIgG1) was administered. Infusion of Fc-Abdeg and IVIG was administered to martins (2 monkeys per group). The animals were administered a volume of 20 ml / kg. Blood samples (3 x 150 μl) were taken 5 min before dosing. ("Before administration"), 5 minutes after the end of infusion, 2 hours, 6 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours Tracer levels were determined by mCD70 binding ELISA. The clinical dose (2 g / kg) of IVIG treatment was compared with the control group and plotted against the pre-treatment levels (Figure 4). In comparison, 70 mg / kg Fc-Abdeg showed significantly enhanced tracer clearance kinetics, More efficient clearance was achieved (>95% tracer clearance in 4 days for Abdeg). for IVIG, ~75% clearance in 7 days).

[0095] Example 2: Effect of afucosylation on the affinity of Fc-Abdeg to human CD16a and mouse CD16-2 effect) The binding affinity of Fc-Abdeg to hCD16a was determined and compared with the afucosylated form (Fc-Abdeg-POT). The same experiment included the Fc-Abdeg variant (Fc-Abdeg), which shows improved affinity for all FcγRs. Specifically, Maxisorp plates were filled with 100 ng / well of NeutrAbdeg-S239D / I332E. The plate was coated with ravidin biotin-binding protein (ThermoScientific, 31000) and incubated at 4°C. The next day, the plates were blocked with PBS + 1% casein for 2 hours at room temperature. Next, biotin-labeled hCD16a (Sino Biological Co., Ltd., 10389-H27H1-B) was added to the 25 100 μl / well of a 0 ng / ml solution (diluted in PBS + 0.1% casein) was added to the plate, and Fc-Ab A concentration gradient (1 μM to 0.005 nM) of either deg or Fc-Abdeg-POT molecules was applied to the cells for 1 h at room temperature. The antibody was incubated for an additional hour after application. Binding to hCD16a was confirmed by HRP binding. Synthetic polyclonal goat anti-human Fc antibody (Jackson ImmunoResearch, 109-035-008) (room temperature Incubate for 1 hour at 100°C, followed by 1 / 50,000 dilution in PBS + 0.1% casein. Detection was performed by adding 1 μl of TMB (SDT Reagent #s TMB) equilibrated to room temperature. The mixture was incubated for 10 minutes, after which 100 μl of 0.5 N H2SO4 was added and the OD450nm was measured. EC50 values ​​were determined using GraphPad Prism software. Experimental results show that defucosylation of the Fc-Abdeg molecule increases its affinity for hCD16a by >30-fold. (EC for Fc-Abdeg-POT) 50 = 13 nM, compared with fucosylated Fc-Abdeg EC 50 As expected, the Fc-Abdeg-S239D / I332E variant showed a significant increase in binding to hCD16a. The binding affinity of wild-type Fc-Abdeg (EC 50 =6 nM).

[0096] Using experimental procedures similar to those described previously, mouse CD16-2 (Sino Biological Co., Ltd., 50036- The binding affinity of the IgG to the IgG antibody (M27H-B) was determined. The results of these experiments, described in Figure 6, also The afucosylated variants exhibited higher affinity compared to wild-type Fc-Abdeg. (EC 50 = 11 nM vs. EC 50 >100 nM). mCD16 of Fc-Abdeg-POT variants relative to wild-type Fc-Abdeg. The fold increase in affinity for β-2 is compared to the fold increase observed for binding to human CD16a. This effect is low compared with the Fc-Abdeg-S239D / I332E variant (EC 50 = 2 nM) was observed. The variant exhibited similar binding to both human and mouse CD16 compared to wild-type Fc-Abdeg. with a fold increase in affinity (EC 50 =2 nM).

[0097] Autoantibodies complexed with autoantigens bind to activated FcγRs, thereby promoting autoimmunity. Autoimmune diseases are partly caused by immunologically mediated inflammation directed against the body's own tissues. This occurs due to the interaction of Fc-Abdeg with autoimmune antibodies and FcγRIII receptors on NK cells. The ability of the compounds to antagonize IL-1 was assessed in two ADCC-based assays.

[0098] First, we used an ADCC reporter bioassay (Promega, G7016) to detect Fc-Abdeg, Fc-Abd The competitive hCD16a binding potency of eg-POT and Fc-Abdeg-S239D / I332E was analyzed. 10,000 CD20-expressing Raji cells ( The target cells were co-incubated with 60,000 hCD16a-expressing Jurkat cells (effector cells). The cells were incubated at 37°C for 6 hours, after which the ADCC activity was assessed. Bioluminescence signals were measured. Luciferase signals were measured at 100 ng / ml in the absence of competitors. These were plotted against the signal obtained with anti-CD20 (see Figure 7). The experiment demonstrated that both Fc-Abdeg-POT and Fc-Abdeg-S239D / I332E effectively inhibited anti-CD20-induced ADCC signals. In contrast, incubation with wild-type Fc-Abdeg efficiently blocked the expression of Fc-Abdeg on Jurkat cells. These results demonstrate that the antibody does not result in competitive binding to hCD16a.

[0099] In the following ADCC assay, anti-hCD70 antibody (27B3-hIgG1) was used with Fc-Abdeg and Fc-Abdeg-POT. Inhibition of the lytic activity of approximately 50,000 hCD16 antibodies was tested as a measure of competitive hCD16 binding. hCD70-expressing U266 cells were cultured in the presence of 50 ng / ml of anti-hCD70 antibody and Fc-Abdeg, Fc-Abdeg-POT, and IVIG. Approximately 300,000 freshly purified PBMCs from healthy donors were added in the presence of a concentration gradient of U266 cells were incubated for 2 days, and subsequent cell lysis was performed to identify a marker specific for U266 cells (CD2 The results of these experiments, shown in Figure 8, demonstrate that anti-CD70 antibodies It efficiently lyses U266 cells, and addition of Fc-Abdeg-POT induced this elimination in a dose-dependent manner. Although this clearance can be attenuated by both wild-type Fc-Abdeg and IVIG, These data indicate that the Fc-Abdeg POT is unable to differentiate between wild-type Fc-Abdeg and IV These results demonstrate that IG possesses enhanced competitive CD16a binding properties compared to IG.

[0100] Example 3: Mouse Acute ITP Model The therapeutic efficacy of Fc-Abdeg, Fc-Abdeg-POT, and Fc-Abdeg-S239D / I332E molecules in acute immune thrombocytopenia Specifically, C57BL / 6 mice were administered I. VIG (20 mg / animal), Fc-Abdeg (1 mg / animal), Fc-Abdeg-POT (1 mg / animal), Fc-Abdeg-S239D The animals were treated with I332E (1 mg / animal) or saline (5 animals / group). After 1 hour, the mice were treated with 5 μg / animal of anti-mammary antibody. The mice were treated with the mouse platelet antibody MWReg30 (Nieswandt et al., Blood 94:684-93 (1999)). Platelet counts were monitored over a 24-hour period. Platelet counts were determined using flow cytometry via anti-CD61 staining, normalized to 0.01%. The results of these experiments described in 9 demonstrate that pretreatment with Fc-Abdeg inhibits MWReg30-induced hematopoietic proliferation. thrombocytopenia was reduced with comparable efficacy compared to IVIG at a 7-fold higher molar dose, and Furthermore, Fc-Abdeg PO Blockade of FcγRs by T and Fc-Abdeg-S239D / I332E has a synergistic beneficial effect in this model This indicates that

[0101] Example 4: Producibility of Fc-Abdeg Fc-Abdeg (containing the Fc domain with SEQ ID NO: 2) was transiently transfected into CHO cells ( Following transfection, high titers of Fc-Abdeg were detected in the supernatant. Fc-Abdeg was injected into CHO GS-XCEED cell line (Lonza, Great Britain) at 200-400 mg / ml. Similar good production profiles were observed when expressed from expression constructs stably integrated into GFP. Stable transfectants produced an average of 3 g / L in a 10 L stirred tank bioreactor. Several clones producing up to 6 g / L of Fc-Abdeg were identified.

[0102] Aggregates and degradation products after Protein A purification of the previously described Fc-Abdeg production runs The manufacturability of Fc-Abdeg was further investigated by analyzing the product. Specifically, 137 μg of Fc-Abdeg Superdex 200 10 / 300 GL gel filtration coupled to an AktaPurifier chromatography system The results of this experiment, shown in Figure 10, show that very small Only a small percentage of Fc-Abdeg aggregates (~0.5%) were observed, whereas Fc-Abdeg degradation products Furthermore, various substances were not detected in the protein A-purified Fc-Abdeg. Even when various stress conditions (freeze-thaw, rotation or temperature stress) are applied, the physicochemical and functional properties remain unchanged. Taken together, these data suggest that demonstrates the excellent manufacturability of Fc-Abdeg.

[0103] Example 5: Dose-escalation study of Fc-Abdeg in cynomolgus monkeys To determine the onset and saturation dose of pharmacokinetic effects, Fc-A was administered in cynomolgus monkeys. A dose-escalation study of bdeg was conducted. For this purpose, cynomolgus monkeys were treated with 1 mg / kg of anti-mouse CD70 hI IgG1 tracer antibody (FR70-hIgG1) was administered by intravenous bolus injection. After 48 hours, the animals Various doses of FC-Abdeg (0.2 mg / kg, 2 mg / kg, 20 mg / kg, or 200 mg / kg) or vehicle were administered to the mice. The infusion was performed within 3 hours and the animals received a volume of 36.36 ml / kg. Each test group consisted of two animals. Five minutes before dosing ("pre-dose"), and after fluid infusion. 5 minutes, 2 hours, 6 hours, 24 hours, 48 ​​hours, 72 hours, 5 days, 7 days, 10 days, and 14 days after the end of Blood samples (3 x 150 μl) were taken at 10 min. Levels of both IgG (see Figure 12) were determined by ELISA and compared to pre-dose levels. The data from the 70 mg / kg dose were added from the experiment described in Figure 4 herein. Administration of 0.2 mg / kg of FC-Abdeg to animals did not significantly affect the clearance rate of the tracer. No effect was observed, and there was no effect on endogenous IgG levels. Pharmacokinetics were evident at 2 mg / kg. This effect plateaued at doses of 20 mg / kg or higher. As a result, IgG in cynomolgus monkeys decreases by up to 55% within 3 to 4 days.

[0104] The binding of FcRn is restricted to the gamma subtype of immunoglobulins and is not associated with other immunoglobulins. This could explain the much longer half-life of the γ subtype compared to the γ subtype. Blockade of the IgG recycling function of FcRn by Fc-Abdeg reduces the levels of endogenous non-IgG immunoglobulins. This characteristic was observed in serum samples from animals treated with 200 mg / kg of Fc-Abdeg. This was demonstrated by measuring the levels of endogenous IgA and IgM (see Figures 13 and 14). ) as described in Example 1, which shows that treatment with Fc-Abdeg does not affect albumin levels. Taken together with the observed results, these data demonstrate specific pharmacokinetic effects of Fc-Abdeg. do.

[0105] Next, the pharmacokinetic profile of Fc-Abdeg was determined by ELISA. As calculated from the IgG antibody (see Figure 5), Fc-Abdeg has a short half-life (estimated half-life). At saturating levels, Fc-Abdeg that is not bound to FcRn binds to itself. Furthermore, the molecular size of Fc-Abdeg is This is close to the cutoff for renal clearance (~60 kDa).

[0106] Example 6: Repeated administration test of Fc-Abdeg in cynomolgus monkeys A single infusion of Fc-Abdeg at a dose of 20 mg / kg resulted in cynomolgus monkeys receiving steroids within approximately 3-4 days. In subsequent experiments, repeated administration of Fc-Abdeg reduced the levels of endogenous IgG in the IgG subpopulation by 55%. We tested whether the levels of α-glucan were further reduced in one group of monkeys. The subjects were given an infusion of the test compound every 24 hours for the first 4 days (days 1, 2, 3, and 4), whereas the subjects were given an infusion of the test compound every 24 hours for the first 4 days (days 1, 2, 3, and 4). The group received the drug every four days (days 1, 5, 9, and 13). For each individual administration, Fc-Abdeg Each test group consisted of two animals. The procedure was identical to that described in Example 1. No obvious pharmacodynamic differences between the two different groups. is observed at day 7 (see Figure 16). Daily Fc-Abdeg during the first 4 days administration is compared to a single infusion of the same dose (dashed line, data from Example 1 overlaid for reference). The administration of Fc-Abdeg once every four days reduced these levels. This results in a larger and longer decline in blood cholesterol. observed in the treatment group.

[0107] The pharmacokinetic profile of Fc-Abdeg was determined (see Figure 7). These results are consistent with findings from the dose escalation study described in Example 5.

[0108] Example 7: Further dose-escalation study of Fc-Abdeg in cynomolgus monkeys To further explore the onset and saturation of the pharmacodynamic effect, Fc-Abdeg was administered in cynomolgus monkeys. A follow-up dose escalation study was conducted. For this purpose, animals were treated with various doses of FC-Abdeg (10 mg / kg, 30 The mice were infused with either 0.1 mg / kg, 50 mg / kg, or 100 mg / kg of the vehicle (PBS). The infusion was administered within 2 hours. Each cohort consisted of four animals (two males and two females). IgG levels were determined by ELISA and plotted against pre-dose levels (see Figure 18). A moderate pharmacodynamic effect was observed at a dose of 10 mg / kg, and this effect was not observed at doses of 30 mg / kg or higher. The rate of IgG clearance, onset of action, and pharmacokinetic profile The results (see Figure 19, see Table 2) are comparable to those reported in Example 5. It was.

[0109] Table 2: Pharmacokinetic properties of Fc-Abdeg in cynomolgus monkeys [Table 2] #: Values ​​obtained from serum analysis of Fc-Abdeg; all other values ​​calculated from toxicokinetic analysis. It was. -: Impossible to calculate or not reasonably interpretable from available data DPF: Dose proportional factor = [AUC 0-t ラスト (x mg / kg) / AUC 0-t ラスト (10 mg / kg)] / [(x mg / k g) / (10 mg / kg)]

[0110] Example 8: Further repeated administration study of Fc-Abdeg in cynomolgus monkeys To further investigate the pharmacodynamic effects of long-term Fc-Abdeg administration, Fc-Abd was administered in cynomolgus monkeys. A follow-up repeated-dose study of FC-Abdeg (3 mg / kg e) was conducted. , 30 mg / kg and 100 mg / kg) or vehicle (PBS) were infused every 48 hours for a total of 15 infusions. The treatment period was followed by a 30-day recovery period. Each cohort consisted of 4 animals. with the exception of the 100 mg / kg cohort (which consisted of 3 animals). Endogenous IgG levels were determined by ELISA and plotted against pre-dose levels (Figure 20). (See Table 1. Mean ± SEM). A clear decrease in IgG levels was observed at all doses tested, This pharmacodynamic effect persisted until the end of the treatment period. After the treatment period, IgG levels remained stable for 10 days. The pharmacokinetic profile is shown in Figure 21.

[0111] Example 9: Long-term administration test of Fc-Abdeg in cynomolgus monkeys Cynomolgus monkeys were infused with an intravenous loading dose of 20 mg / kg of FC-Abdeg, and the dose was administered 24 hours after the first dose. Patients were further treated with daily subcutaneous administration of 1, 3, or 5 mg / kg Fc-Abdeg starting from day 1 and continuing for 12 days. Each test group consisted of two animals. IgG levels were determined by ELISA and The data are plotted against pre-treatment levels (see Figure 22). A study was presented for one cynomolgus monkey from the 3 mg / kg cohort who continued treatment. The monkeys received an intravenous loading dose of 20 mg / kg of FC-Abdeg, and 24 hours after the first dose, Administer daily subcutaneous administration of 3 mg / kg Fc-Abdeg for 28 days, followed by an additional 32 days. A treatment-free period was then allowed between treatments (see Figure 23). IgG levels increased to a maximum of 60% of the baseline level. After the treatment period, IgG levels decreased within 2 weeks. Levels returned to baseline levels. The present application provides the following inventions. (Configuration 1) A method for reducing serum levels of an Fc-containing agent in a subject, comprising administering to the subject an effective amount of and administering an isolated FcRn antagonist comprising a resistant Fc region or an FcRn-binding fragment thereof. and wherein the Fc domain of the variant Fc region comprises EU positions 252, 254, 256, 433, 434, and 436. and wherein the FcRn antagonist comprises the amino acids Y, T, E, K, F, and Y, respectively, and The method, wherein the compound is administered at a dose of about 0.2 to about 200 mg / kg. (Configuration 2) A method for reducing serum levels of an Fc-containing agent in a subject, comprising administering to the subject an effective amount of and administering an isolated FcRn antagonist comprising a resistant Fc region or an FcRn-binding fragment thereof. and wherein the Fc domain of the variant Fc region comprises EU positions 252, 254, 256, 433, 434, and 436. and wherein the FcRn antagonist comprises the amino acids Y, T, E, K, F, and Y, respectively, and The method, wherein the compound is administered at least twice every 20 days. (Configuration 3) the FcRn antagonist is administered to the subject once every 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 days; 3. The method of claim 2, wherein the compound is administered at a frequency of 100 mg / kg / day. (Configuration 4) 3. The method of claim 2, wherein the FcRn antagonist is administered to the subject once every four days. Law. (Configuration 5) 3. The method of claim 2, wherein the FcRn antagonist is administered to the subject once every 7 days. Law. (Configuration 6) The FcRn antagonist is administered to the subject at 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32 4. The method of claim 2 or 3, wherein 13, 14, 15, 16, 17, 18, 19, or 20 doses are administered. (Configuration 7) A method for reducing serum levels of an Fc-containing agent in a subject, comprising administering to the subject an effective amount of and administering an isolated FcRn antagonist comprising a resistant Fc region or an FcRn-binding fragment thereof. and wherein the Fc domain of the variant Fc region comprises EU positions 252, 254, 256, 433, 434, and 436. and wherein the FcRn antagonist comprises the amino acids Y, T, E, K, F, and Y, respectively, and The method is administered once every 48 hours for four weeks. (Configuration 8) The FcRn antagonist is administered to the subject at a dose of about 0.2 to about 200 mg / kg. 7. The method according to any one of items 2 to 6. (Configuration 9) The FcRn antagonist is administered to the subject in an amount of about 0.2, 1, 2, 3, 5, 10, 20, 25, 30, 50, 70, 80, 90, 100, 100, 150, 160, 170, 180, 190, 210, 220, 230, 240, 250, 2 9. The method of any one of claims 1 to 8, wherein the compound is administered at a dose of 100, 100, or 200 mg / kg. (Configuration 10) 10. The method of any one of claims 1 to 9, wherein the FcRn antagonist is administered to the subject at a dose of about 10 mg / kg. 1. The method according to any one of the preceding items. (Configuration 11) 11. The method of claim 1, wherein the FcRn antagonist is administered to the subject at a dose of about 20 mg / kg. 10. The method of any one of claims 1 to 9. (Configuration 12) 12. The method of any one of claims 1 to 11, wherein the FcRn antagonist is administered to the subject at a dose of about 25 mg / kg. 10. The method of any one of claims 1 to 9. (Configuration 13) 13. The method according to any one of aspects 1 to 12, wherein the FcRn antagonist is administered intravenously. (Configuration 14) 14. The method according to any one of aspects 1 to 13, wherein the FcRn antagonist is administered subcutaneously. (Configuration 15) The FcRn antagonist is administered to the subject in two or more doses, wherein A first dose of the compound is administered intravenously, followed by one or more subsequent doses administered subcutaneously. The method according to any one of the preceding claims. (Configuration 16) 16. The method according to any one of Aspects 1 to 15, wherein the FcRn antagonist does not contain an antibody variable region. Law. (Configuration 17) 17. The method according to any one of aspects 1 to 16, wherein the FcRn antagonist does not contain a CH1 domain. Law. (Configuration 18) 18. The method according to any one of claims 1 to 17, wherein the FcRn antagonist does not contain a free cysteine ​​residue. How to post. (Configuration 19) 19. The method according to any one of aspects 1 to 18, wherein the variant Fc region is an IgG Fc region. (Configuration 20) 20. The method according to any one of aspects 1 to 19, wherein the variant Fc region is an IgG1 Fc region. (Configuration 21) The amino acid sequence of the Fc domain of the variant Fc region is set forth in SEQ ID NO: 1, 2, or 3. 21. The method according to any one of aspects 1 to 20, comprising the amino acid sequence of (Configuration 22) the amino acid sequence of the Fc domain of the variant Fc region is the amino acid sequence set forth in SEQ ID NO: 1 22. The method according to any one of aspects 1 to 21, wherein the sequence (Configuration 23) The FcRn antagonist comprises a variant Fc region, and the Fc domain of the variant Fc region The amino acid sequence of the amino acid sequence of SEQ ID NO: 1, 2, or 3 is selected from the group consisting of: The method according to any one of the preceding claims. (Configuration 24) the variant Fc region has an affinity for Fcγ receptors that is higher than that of a wild-type IgG1 Fc region 24. The method according to any one of aspects 1 to 23, wherein the antibody has increased affinity for an Fcγ receptor. (Configuration 25) 25. Any of configurations 1 to 24, wherein the variant Fc region has increased affinity for CD16a. or the method described in paragraph 1. (Configuration 26) the Fc domain of the variant Fc region does not contain an N-linked glycan at EU position 297. The method according to any one of items 1 to 25. (Configuration 27) The FcRn antagonist comprises a plurality of FcRn antagonist molecules, At least 50% (optionally at least 60, 70, 80, 90, 95, or 99%) of the agonist molecules , a variant Fc region comprising an afucosylated N-linked glycan at EU position 297 or its FcRn binding 26. The method of any one of embodiments 1 to 25, comprising a fragment. (Configuration 28) The FcRn antagonist comprises a plurality of FcRn antagonist molecules, At least 50% (optionally at least 60, 70, 80, 90, 95, or 99%) of the agonist molecules , a variant Fc containing an N-linked glycan with a bisecting GlcNac at EU position 297 26. The method according to any one of aspects 1 to 25, comprising the region or an FcRn-binding fragment thereof. (Configuration 29) Any of configurations 1 to 28, wherein the variant Fc region is linked to a half-life extender. or the method described in paragraph 1. (Configuration 30) the half-life extender is polyethylene glycol or human serum albumin; The method according to any one of aspects 1 to 29. (Configuration 31) 31. The method according to any one of aspects 1 to 30, wherein the Fc-containing agent is an antibody or an immunoadhesin. Law. (Configuration 32) 32. The method according to any one of aspects 1 to 31, wherein the Fc-containing agent is a therapeutic agent or a diagnostic agent. (Configuration 33) 33. The method according to any one of aspects 1 to 32, wherein the Fc-containing agent is an imaging agent. (Configuration 34) 34. The method according to any one of aspects 1 to 33, wherein the Fc-containing agent is an antibody-drug conjugate. . (Configuration 35) 35. The method of any one of aspects 1 to 34, wherein the Fc-containing agent is a pathogenic antibody. (Configuration 36) 36. The method according to any one of aspects 1 to 35, wherein the Fc-containing agent is an autoantibody. (Configuration 37) the subject has an antibody-mediated disease or disorder, and the FcRn antagonist is administered to the subject. 37. The method according to any one of aspects 1 to 36, wherein the disease or disorder is improved by administering the (Configuration 38) 38. The method of claim 37, wherein the antibody-mediated disease or disorder is an autoimmune disease. (Configuration 39) The autoimmune disease is selected from the group consisting of allogeneic islet transplant rejection, alopecia areata, ankylosing spondylitis, anti-inflammatory drugs, and the like. Phospholipid syndrome, autoimmune Addison's disease, Alzheimer's disease, antineutrophil cytoplasmic autoantibodies (ANCA), autoimmune disease of the adrenal gland, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune Myocarditis, autoimmune neutropenia, autoimmune oophoritis and orchitis, autoimmune thrombocytopenia Hypothyroidism, autoimmune urticaria, Behçet's disease, bullous pemphigoid, cardiomyopathy, Castleman's disease syndrome, celiac disease-dermatitis, chronic fatigue immune deficiency syndrome, chronic inflammatory demyelinating polyneuropathy (C IDP), Churg-Strauss syndrome, cicatricial pemphigoid, CREST syndrome, cold agglutinin disease, Crohn's disease, dermatomyositis, dilated cardiomyopathy, discoid lupus erythematosus, epidermolysis bullosa acquisita, Atopic mixed cryoglobulinemia, factor VIII deficiency, fibromyalgia-fibromyositis, glomerular nephropathy inflammation, Graves' disease, Guillain-Barré syndrome, Goodpasture syndrome, graft-versus-host disease (GVHD) , Hashimoto's thyroiditis, hemophilia A, idiopathic membranous neuropathy, idiopathic pulmonary fibrosis, idiopathic platelet Inflammatory purpura (ITP), IgA neuropathy, IgM polyneuropathy, immune-mediated hematopoietic Platelet deficiency, juvenile arthritis, Kawasaki disease, lichen planus, lichen sclerosus, lupus erythematosus, Meniere's disease Malignant leukemia, mixed connective tissue disease, mucous membrane pemphigoid, multiple sclerosis, type 1 diabetes, multifocal motor neuron syndrome Myasthenia gravis (MMN), paraneoplastic bullous pemphigoid, pemphigoid gestationis, pemphigoid vulgaris Smallpox, pemphigus foliaceus, pernicious anemia, polyarteritis nodosa, polychondritis, polyglandular syndrome, rheumatoid arthritis Polymyalgia machinosum, polymyositis and dermatomyositis, primary agammaglobulinemia, primary cholesteatoma Juvenile cirrhosis, psoriasis, psoriatic arthritis, relapsing polychondritis, Raynaud's phenomenon, Reiter's syndrome , rheumatoid arthritis, sarcoidosis, scleroderma, Sjogren's syndrome, solid organ transplant rejection Asymptomatic reaction, stiff man syndrome, systemic lupus erythematosus, Takayasu's arteritis, toxic epidermal necrolysis syndrome (TEN), Stevens-Johnson syndrome (SJS), temporal arteritis / giant cell arteritis, Thrombotic thrombocytopenic purpura, ulcerative colitis, uveitis, dermatitis herpetiformis, vasculitis, anti-inflammatory selected from the group consisting of neutrophil cytoplasmic antibody-associated vasculitis, vitiligo, and Wegener's granulomatosis; 39. The method of claim 38. (Configuration 40) The disease or disorder is treated with intravenous immunoglobulin (IVIG), plasma exchange therapy, and / or immunotherapy. 38. The method of claim 37, wherein the treatment is possible using an adsorption method. (Configuration 41) 39. The method of claim 38, wherein the autoimmune disease is an autoimmune channelopathy. (Configuration 42) The channelopathy may be autoimmune limbic encephalitis, epilepsy, neuromyelitis optica, Lambert-I. Peterson's myasthenic syndrome, myasthenia gravis, anti-N-methyl-D-aspartate (NMDA) receptor encephalitis , anti-α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor brain Pediatric autoimmune psychiatric disorders associated with inflammatory bowel disease, Morvan's syndrome, neuromyotonia, and streptococcal infections neuropathy (PANDAS), and glycine receptor antibody-associated disorder. 41. The method described in 41. (Configuration 43) 38. The method of claim 37, wherein the antibody-mediated disease or disorder is hyperglobulinemia. (Configuration 44) The FcRn antagonist is administered to the subject simultaneously or sequentially with an additional therapeutic agent. The method according to any one of aspects 1 to 43, (Configuration 45) 45. The method of claim 44, wherein the additional therapeutic agent is an anti-inflammatory agent. (Configuration 46) 45. The method of claim 44, wherein the additional therapeutic agent is a leukoreducing agent. (Configuration 47) 46. ​​The method of claim 45, wherein the leukocyte depleting agent is a B cell depleting agent. (Configuration 48) 48. The method of claim 47, wherein the B cell depleting agent is an antibody. (Configuration 49) The B cell depleting agent is selected from the group consisting of CD10, CD19, CD20, CD21, CD22, CD23, CD24, CD37, CD53, and CD70 , CD72, CD74, CD75, CD77, CD79a, CD79b, CD80, CD81, CD82, CD83, CD84, CD85, or 49. The method of claim 48, wherein the antibody specifically binds to CD86. (Configuration 50) The additional therapeutic agent is rituximab, daclizumab, basiliximab, muronomab-CD3 , infliximab, adalimumab, omalizumab, efalizumab, natalizumab, Silizumab, eculizumab, golimumab, canakinumab, ustekinumab, belimumab, 50. The method of claim 49, wherein the method is a combination thereof. (Configuration 51) 51. The method according to any one of aspects 1 to 50, wherein the subject is a human or a cynomolgus monkey.

Claims

1. 1. A pharmaceutical composition for use in a method for reducing serum levels of IgG autoantibodies in a subject, the method comprising: the variant IgG1 Fc region consists of two Fc domains that form a dimer; the Fc domain of the variant IgG1 Fc region comprises amino acids Y, T, E, K, F, and Y at EU positions 252, 254, 256, 433, 434, and 436, respectively; the FcRn antagonist is administered to the subject at a dose of 2 to 200 mg / kg; and The pharmaceutical composition, wherein the FcRn antagonist is administered to the subject twice every 20 days.

2. The pharmaceutical composition of claim 1, wherein the FcRn antagonist is administered intravenously.

3. The pharmaceutical composition of claim 1, wherein the FcRn antagonist is administered subcutaneously.

4. The pharmaceutical composition of any one of claims 1 to 3, wherein the FcRn antagonist is administered once every 7, 8, 9, or 10 days.

5. The pharmaceutical composition of any one of claims 1 to 4, wherein the FcRn antagonist is administered once every seven days.

6. The pharmaceutical composition of any one of claims 1 to 4, wherein the FcRn antagonist is administered once every 10 days.

7. 7. The pharmaceutical composition of any one of claims 1 to 6, wherein the variant Fc region is linked to a half-life extender, and the half-life extender is polyethylene glycol or human serum albumin.

8. The pharmaceutical composition of any one of claims 1 to 7, wherein the subject has an antibody-mediated disease or disorder.

9. The pharmaceutical composition of any one of claims 1 to 8, wherein the subject has an autoimmune disease.

10. The autoimmune disease may be selected from the group consisting of allogeneic islet graft rejection, alopecia areata, ankylosing spondylitis, antiphospholipid syndrome, autoimmune Addison's disease, Alzheimer's disease, antineutrophil cytoplasmic autoantibodies (ANCA), autoimmune diseases of the adrenal gland, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune myocarditis, autoimmune neutropenia, autoimmune oophoritis and orchitis, autoimmune thrombocytopenia, autoimmune urticaria, Behcet's disease, bullous pemphigoid, cardiomyopathy, Castleman's disease, and parsley. Achon disease-dermatitis, chronic fatigue immune deficiency syndrome, chronic inflammatory demyelinating polyneuropathy (CIDP), Churg-Strauss syndrome, cicatricial pemphigoid, CREST syndrome, cold agglutinin disease, Crohn's disease, dermatomyositis, dilated cardiomyopathy, discoid lupus erythematosus, epidermolysis bullosa acquisita, essential mixed cryoglobulinemia, factor VIII deficiency, fibromyalgia-fibromyositis, glomerulonephritis, Graves' disease, Guillain-Barré syndrome, Goodpasture's syndrome, graft-versus-host disease (GVHD), Hashimoto's thyroiditis, hemophilia A, idiopathic membranous neuropathy, idiopathic pulmonary fibrosis, idiopathic thrombocytopenic purpura (ITP), IgA neuropathy, IgM polyneuropathy, immune-mediated thrombocytopenia, juvenile arthritis, Kawasaki disease, lichen planus, lichen sclerosus, lupus erythematosus, Meniere's disease, mixed connective tissue disease, mucous membrane pemphigoid, multiple sclerosis, type 1 diabetes, multifocal motor neuropathy (MMN), myasthenia gravis, paraneoplastic bullous pemphigoid, pemphigoid of pregnancy, pemphigoid vulgaris 10. The pharmaceutical composition of claim 9, wherein the inflammatory bowel disease is selected from the group consisting of acne, pemphigus foliaceus, pernicious anemia, polyarteritis nodosa, polychondritis, polyglandular syndrome, polymyalgia rheumatica, polymyositis and dermatomyositis, primary agammaglobulinemia, primary biliary cirrhosis, psoriasis, psoriatic arthritis, relapsing polychondritis, Raynaud's phenomenon, Reiter's syndrome, rheumatoid arthritis, sarcoidosis, scleroderma, Sjogren's syndrome, solid organ transplant rejection, stiff man syndrome, systemic lupus erythematosus, Takayasu's arteritis, toxic epidermal necrolysis (TEN), Stevens-Johnson syndrome (SJS), temporal arteritis / giant cell arteritis, thrombotic thrombocytopenic purpura, ulcerative colitis, uveitis, dermatitis herpetiformis vasculitis, antineutrophil cytoplasmic antibody-associated vasculitis, vitiligo, and Wegener's granulomatosis.

11. 10. The pharmaceutical composition of claim 9, wherein the autoimmune disease is an autoimmune channelopathy.

12. 12. The pharmaceutical composition of claim 11, wherein the channelopathy is selected from the group consisting of autoimmune limbic encephalitis, epilepsy, neuromyelitis optica, Lambert-Eaton myasthenic syndrome, myasthenia gravis, anti-N-methyl-D-aspartate (NMDA) receptor encephalitis, anti-α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor encephalitis, Morvan's syndrome, neuromyotonia, pediatric autoimmune neuropsychiatric disorder associated with streptococcal infection (PANDAS), and glycine receptor antibody-associated disorder.

13. The pharmaceutical composition according to any one of claims 1 to 12, wherein the FcRn antagonist is administered to the subject simultaneously or sequentially with an additional therapeutic agent.

14. 14. The pharmaceutical composition of claim 13, wherein the additional therapeutic agent is an anti-inflammatory agent or a leukoreducing agent.

15. 15. The pharmaceutical composition of claim 14, wherein the additional therapeutic agent is a leukocyte-depleting agent, and the leukocyte-depleting agent is a B-cell-depleting agent.

16. 16. The pharmaceutical composition of claim 15, wherein the B cell depleting agent is an antibody.

17. The pharmaceutical composition of claim 16, wherein the B cell depleting agent is an antibody that specifically binds to CD10, CD19, CD20, CD21, CD22, CD23, CD24, CD37, CD53, CD70, CD72, CD74, CD75, CD77, CD79a, CD79b, CD80, CD81, CD82, CD83, CD84, CD85, or CD86.

18. 14. The pharmaceutical composition of claim 13, wherein the additional therapeutic agent is rituximab, daclizumab, basiliximab, muromonab-CD3, infliximab, adalimumab, omalizumab, efalizumab, natalizumab, tocilizumab, eculizumab, golimumab, canakinumab, ustekinumab, belimumab, or a combination thereof.

19. The pharmaceutical composition according to any one of claims 1 to 18, wherein the subject is a human or a cynomolgus monkey.

20. The pharmaceutical composition of any one of claims 1 to 19, wherein the subject is a human.

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