PHARMACEUTICAL FORMULATIONS OF FcRn INHIBITORS SUITABLE FOR SUBCUTANEOUS ADMINISTRATION

Abstract

To provide improved formulations and methods of administration of FcRn inhibitors for use in the treatment of autoimmune diseases.SOLUTION: Provided are various aqueous formulations of the neonatal Fc receptor (FcRn) antagonist ARGX-113, including formulations useful as pharmaceutical compositions, methods for their preparation, devices comprising the various formulations, and uses thereof. In certain embodiments the formulations are suitable and useful for administration of ARGX-113 to a human subject. In certain embodiments the formulations are suitable and useful for subcutaneous administration of ARGX-113 to a human subject. The formulations can be used in the treatment of any disease that would benefit from inhibition of FcRn-mediated antibody recycling. Such diseases can include any one or more of various antibody-mediated autoimmune diseases, including, for example and without limitation, myasthenia gravis (MG) and immune thrombocytopenia (ITP).SELECTED DRAWING: Figure 1

Description

[Background technology]

[0001] (background) Immunoglobulin gamma (IgG) antibodies are used to treat autoimmune diseases, inflammatory diseases, and conditions where the pathology is IgG antibodies. The pathology of many disorders, including those characterized by their overexpression (e.g., hypergammaglobulinemia) (See, e.g., Junghans, Immunol Res. 16(1):29 (1997) reference).

[0002] The half-life of IgG in serum is long compared to the serum half-lives of other plasma proteins (Roop Enian et al., J Immunol. 170:3528 (2003); Junghans and Anderson, Proc. Natl. Acad. Sci. USA 93:5512(1996)). This long half-life is due in part to the neonatal Fc receptor (FcRn) FcRn is a receptor that originally acts as a transport receptor for maternal IgG to the newborn. It has been characterized as a cytotoxic agent, but also functions to protect IgG from degradation in adults. FcRn binds pinocytosed IgG and recycles it back to the extracellular compartment. This recycling protects IgG from trafficking to the degradative lysosomes. The IgG / FcRn interaction is driven by pH-dependent binding of IgG at extracellular physiological pH. is stronger at the acidic pH of the endosome than at

[0003] When the serum concentration of IgG reaches a level that exceeds the available FcRn molecules, unbound IgG is cleaved As a result, they are not protected from degradation mechanisms and have a reduced serum half-life. Thus, inhibition of IgG binding to FcRn may inhibit I by preventing endosomal recycling of IgG. Therefore, agents that antagonize IgG binding to FcRn, e.g. For example, the compounds may be useful in regulating, treating, or preventing antibody-mediated disorders, such as autoimmune diseases and inflammatory diseases. Some of these diseases may be due, at least in part, to human donor-derived genes. Many of these autoimmune diseases are currently treated with intravenous infusion of immune IgG (IVIg). Because most infections are chronic, affected individuals may need to use IVIg and / or other treatments to manage their disease. Repeated administration of the appropriate therapy may be required.

[0004] In another approach, a blocking antibody against FcRn was developed to inhibit IgG Fc binding to FcRn. (See, for example, WO 2002 / 043658.) Peptides have also been identified (see, e.g., US 6,212,022 and US 8,101,186). A variant with enhanced FcRn binding and reduced pH dependence that antagonizes FcRn binding to IgG. Full-length IgG antibodies that contain specific Fc receptors have also been identified (see, e.g., U.S. Pat. No. 8,163,881 and Vaccaro et al. (See, e.g., Nat Biotechnol. 23(10): 1283-1288 (2005)).

[0005] Recently, another F, named efgaltigimod, which is a modified version of the human IgG1 Fc fragment, has been reported. A cRn inhibitor (also known as ARGX-113) has been developed, the entire contents of which are incorporated herein by reference. See WO 2015 / 100299, incorporated herein by reference. ARGX-113 is currently being used to treat myasthenia gravis (M It is currently undergoing clinical trials in several autoimmune diseases, including immune thrombocytopenia (ITP) and rheumatoid arthritis (HR). This is currently being done.

[0006] Improved formulations and administration of FcRn inhibitors for use in the treatment of autoimmune diseases There remains a need for methods of providing Summary of the Invention

[0007] (overview) Disclosed herein are various formulations of ARGX-113, including formulations useful as pharmaceutical compositions. , methods for their preparation, devices containing the various formulations, and uses thereof. Thus, the formulation is suitable and useful for administration of ARGX-113 to a human subject. The formulation is suitable and useful for subcutaneous administration of ARGX-113 to a human subject. It can be used in the treatment of any disease that benefits from inhibition of antibody recycling. Such diseases include, but are not limited to, myasthenia gravis (MG) and immune thrombocytopenia. The present invention may include any one or more of a variety of antibody-mediated autoimmune diseases, including immune thrombocytopenic purpura (ITP). This can be done.

[0008] One aspect of the present invention is an isolated neonatal Fc receptor (FcRn) antagonist at about 100 to 300 mg / mL. The mixture was diluted with 20-60 mM histidine / histidine HCl, 0-70 mM sucrose, 0-150 mM NaCl, 0-250 mM mM arginine HCl, 0% to 0.05% (w / v) polysorbate 20 or polysorbate 80, 0 to 15 mM L -methionine, pH 6.0 to 6.5, wherein the isolated FcRn antagonist is The antigen comprises a variant Fc region, wherein the variant Fc region forms a homodimer. and wherein the amino acid sequence of each of the Fc domains is the sequence It consists of sequence number 1, sequence number 2, or sequence number 3.

[0009] In one embodiment, the aqueous formulation contains about 100-300 mg / mL of isolated neonatal Fc receptor (FcRn ) The antagonist was added to a 20-60 mM histidine / histidine HCl, 0-70 mM sucrose, approximately 100 mM NaCl, 0% to 0.05% (w / v) polysorbate 20 or polysorbate 80, 0 to 15 mM L-methionine and pH 6.0 to 6.5, wherein the isolated FcRn antagonist comprises a variant Fc domain. wherein the variant Fc region is composed of two Fc domains that form a homodimer. wherein the amino acid sequence of each of the Fc domains is selected from SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3. It consists of column number 3.

[0010] In one embodiment, the aqueous formulation contains about 100-300 mg / mL of isolated neonatal Fc receptor (FcRn ) The antagonist was added to a 20-60 mM histidine / histidine HCl, 0-70 mM sucrose, 100-250 mM mM arginine HCl, 0% to 0.05% (w / v) polysorbate 20 or polysorbate 80, 0 to 15 mM L -methionine, pH 6.0 to 6.5, wherein the isolated FcRn antagonist is wherein the variant Fc region comprises two Fc domains that form a homodimer. domains, wherein the amino acid sequence of each of the Fc domains is SEQ ID NO:1, SEQ ID NO:2, No. 2, or No. 3.

[0011] In one embodiment, the aqueous formulation contains about 100-300 mg / mL of isolated neonatal Fc receptor (FcRn ) The antagonist was added in a concentration of 20-60 mM histidine / histidine HCl, 100-250 mM arginine HCl, 0 %~0.05% (w / v) polysorbate 20 or polysorbate 80, 0~15mM L-methionine, pH 6. 0 to 6.5, wherein the isolated FcRn antagonist comprises a variant Fc region. wherein the variant Fc region consists of two Fc domains that form a homodimer; wherein the amino acid sequence of each of the Fc domains is SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3. It consists of:

[0012] In each of the above aspects and embodiments, in certain embodiments, the aqueous formulation comprises 20 or Contains 50mM histidine.

[0013] In each of the above aspects and embodiments, in certain embodiments, the aqueous formulation comprises 0.02% Contains ~0.04% (w / v) polysorbate 20 or polysorbate 80.

[0014] In each of the above aspects and embodiments, in certain embodiments, the aqueous formulation comprises 0 or 1 Contains 0 mM L-methionine.

[0015] In each of the above aspects and embodiments, in certain embodiments, the pH is 6.0 or 6.5. be.

[0016] In each of the foregoing aspects and embodiments, in certain embodiments, each of the Fc domains The amino acid sequence consists of SEQ ID NO:1.

[0017] One aspect of the present invention is an isolated neonatal Fc receptor (FcRn) antagonist at about 100 to 200 mg / mL. The solution was prepared in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.02% to 0.04% ethanol. % (w / v) polysorbate 20 or polysorbate 80, pH 6.0, The isolated FcRn antagonist comprises a variant Fc region, wherein the variant The Fc region of a serogroup is composed of two Fc domains that form a homodimer, where the Fc domains Each of the amino acid sequences consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0018] In one embodiment, the aqueous formulation contains about 100-200 mg / mL of ARGX-113 in 20 mM histidine / hybrid. Styridine HCl, 60 mM sucrose, 100 mM NaCl, and 0.02% to 0.04% (w / v) polysorbate 20 or polysorbate 80, pH 6.0, wherein ARGX-113 comprises a variant Fc region. and an isolated FcRn antagonist comprising: The antibody comprises two Fc domains forming an Fc domain having an amino acid sequence similar to that of the Fc domain. It consists of column number 1.

[0019] One embodiment of the present invention is a method for the preparation of 150 mg / mL ARGX-113 in 20 mM histidine / histidine HCl, 60 mM sucrose, The aqueous formulation contained 100 mM glucose, 100 mM NaCl, and 0.04% (w / v) polysorbate 20, pH 6.0. wherein ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc region. wherein the variant Fc region consists of two Fc domains that form a homodimer; Here, the amino acid sequence of each of the Fc domains consists of SEQ ID NO:1.

[0020] One embodiment of the present invention is a method for the preparation of 165 mg / mL ARGX-113 in 20 mM histidine / histidine HCl, 60 mM sucrose, An aqueous formulation containing 100 mM glucose, 100 mM NaCl, and approximately 0.04% (w / v) polysorbate 20, pH 6.0. wherein ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc region. wherein the variant Fc region consists of two Fc domains that form a homodimer. , wherein the amino acid sequence of each of the Fc domains consists of SEQ ID NO:1.

[0021] One embodiment of the present invention is a method for the preparation of 175 mg / mL ARGX-113 in 20 mM histidine / histidine HCl, 60 mM sucrose, The aqueous formulation contained 100 mM glucose, 100 mM NaCl, and 0.04% (w / v) polysorbate 20, pH 6.0. wherein ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc region. wherein the variant Fc region consists of two Fc domains that form a homodimer; Here, the amino acid sequence of each of the Fc domains consists of SEQ ID NO:1.

[0022] One embodiment of the present invention is a method for the preparation of 180 mg / mL ARGX-113 in 20 mM histidine / histidine HCl, 60 mM sucrose, The aqueous formulation contained 100 mM glucose, 100 mM NaCl, and 0.04% (w / v) polysorbate 20, pH 6.0. wherein ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc region. wherein the variant Fc region consists of two Fc domains that form a homodimer; Here, the amino acid sequence of each of the Fc domains consists of SEQ ID NO:1.

[0023] One embodiment of the present invention is a suspension of 200 mg / mL ARGX-113 in water, 20 mM histidine / histidine HCl, 60 mM Aqueous formulation containing sucrose, 100 mM NaCl, and 0.04% (w / v) polysorbate 20, pH 6.0 wherein ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc region. wherein the variant Fc region comprises two Fc domains that form a homodimer. wherein the amino acid sequence of each of the Fc domains consists of SEQ ID NO:1.

[0024] One aspect of the present invention is an isolated neonatal Fc receptor (FcRn) antagonist at about 100 to 200 mg / mL. The solution was diluted with 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and and 0.02% to 0.04% (w / v) polysorbate 20 or polysorbate 80, pH 6.0 an aqueous formulation, wherein the isolated FcRn antagonist comprises a variant Fc region. wherein the variant Fc region consists of two Fc domains that form a homodimer; wherein the amino acid sequence of each of the Fc domains is SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3. It consists of:

[0025] In one embodiment, the aqueous formulation contains about 100-200 mg / mL of ARGX-113 in 20 mM histidine / hybrid. Styridine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.02% to 0.04% (w / v) polysorbate 20 or polysorbate 80, pH 6.0, wherein A RGX-113 is an isolated FcRn antagonist consisting of a variant Fc region, wherein: The variant Fc region is composed of two Fc domains that form a homodimer, wherein the Fc The amino acid sequence of each of the domains consists of SEQ ID NO:1.

[0026] One embodiment of the present invention is a method for administering 165 mg / mL of an isolated neonatal Fc receptor (FcRn) antagonist to a patient in need thereof. mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0 0.04% (w / v) polysorbate 20, pH 6.0, wherein the isolated Fc The Rn antagonist comprises a variant Fc region, wherein the variant Fc region is a homozygous Fc region. It consists of two Fc domains that form a dimer, wherein the amino acid sequence of each of the Fc domains is The string consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0027] In one embodiment, the aqueous formulation contains 165 mg / mL ARGX-113 in 20 mM histidine / histidine. HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.04% (w / v) polysorbate and 20, pH 6.0, wherein ARGX-113 is an isolated Fc antibody comprising a variant Fc region. Rn antagonist, wherein the variant Fc region comprises two Fs that form a homodimer. and c domains, wherein the amino acid sequence of each of the Fc domains consists of SEQ ID NO:1. .

[0028] One embodiment of the present invention is a method for the preparation of 175 mg / mL ARGX-113 in 20 mM histidine / histidine HCl, 60 mM sucrose, in 100 mM NaCl, 10 mM L-methionine, and 0.03% (w / v) polysorbate 20, pH 6.0 wherein ARGX-113 is an isolated FcRn comprising a variant Fc region. antagonist, wherein the variant Fc region comprises two Fc domains that form a homodimer. domains, wherein the amino acid sequence of each of the Fc domains consists of SEQ ID NO:1. .

[0029] One embodiment of the present invention is a method for the preparation of 180 mg / mL ARGX-113 in 20 mM histidine / histidine HCl, 60 mM sucrose, in 100 mM NaCl, 10 mM L-methionine, and 0.03% (w / v) polysorbate 20, pH 6.0 wherein ARGX-113 is an isolated FcRn comprising a variant Fc region. antagonist, wherein the variant Fc region comprises two Fc domains that form a homodimer. domains, wherein the amino acid sequence of each of the Fc domains consists of SEQ ID NO:1. .

[0030] One embodiment of the present invention is a method for the preparation of 200 mg / mL ARGX-113 in 20 mM histidine / histidine HCl, 60 mM sucrose, in 100 mM NaCl, 10 mM L-methionine, and 0.03% (w / v) polysorbate 20, pH 6.0 wherein ARGX-113 is an isolated FcRn comprising a variant Fc region. antagonist, wherein the variant Fc region comprises two Fc domains that form a homodimer. domains, wherein the amino acid sequence of each of the Fc domains consists of SEQ ID NO:1. .

[0031] One aspect of the present invention is an isolated neonatal Fc receptor (FcRn) antagonist at about 100 to 200 mg / mL. The mixture was diluted with 50 mM histidine / histidine HCl, 60 mM sucrose, 150 mM arginine HCl, and 0.02 0.04% (w / v) polysorbate 20 or polysorbate 80, pH 6.0. wherein the isolated FcRn antagonist consists of a variant Fc region, The variant Fc region is composed of two Fc domains that form a homodimer, wherein the Fc The amino acid sequence of each of the domains consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0032] In one embodiment, the aqueous formulation contains about 100-200 mg / mL of ARGX-113 in 50 mM histidine / hybrid. Stidine HCl, 60 mM sucrose, 100 mM arginine HCl, and 0.02% to 0.04% (w / v) Polysol and polysorbate 20 or polysorbate 80, pH 6.0, wherein ARGX-113 is a variant Fc region. and an isolated FcRn antagonist comprising a variant Fc region, The Fc domain comprises a main homodimer, and the amino acid sequence of each of the Fc domains is selected from SEQ ID NO: 1 and SEQ ID NO: 2. Or it will be.

[0033] One aspect of the present invention is an isolated neonatal Fc receptor (FcRn) antagonist at about 100 to 200 mg / mL. The mixture was supplemented with 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM arginine HCl, 10 mM L- Methionine, and 0.02% to 0.04% (w / v) polysorbate 20 or polysorbate 80, pH 6.0 wherein the isolated FcRn antagonist is a variant Fc region. wherein the variant Fc region is composed of two Fc domains that form a homodimer. wherein the amino acid sequence of each of the Fc domains is selected from SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3. It consists of column number 3.

[0034] In one embodiment, the aqueous formulation contains about 100-200 mg / mL of ARGX-113 in 20 mM histidine / hybrid. Styridine HCl, 60 mM sucrose, 100 mM arginine HCl, 10 mM L-methionine, and 0.02% 0.04% (w / v) polysorbate 20 or polysorbate 80, pH 6.0, wherein ARGX-11 3 is an isolated FcRn antagonist consisting of a variant Fc region, The ant Fc region consists of two Fc domains that form a homodimer, where the Fc domains The amino acid sequence of each of the peptides consists of SEQ ID NO:1.

[0035] One embodiment of the present invention is a method for the preparation of 175 mg / mL ARGX-113 in 20 mM histidine / histidine HCl, 60 mM sucrose, sucrose, 100 mM arginine HCl, 10 mM L-methionine, and 0.03% (w / v) polysorbate 20, and an aqueous formulation comprising, at pH 6.0, an isolated antibody comprising a variant Fc region. and an FcRn antagonist, wherein the variant Fc region forms a homodimer. and two Fc domains, wherein the amino acid sequence of each of the Fc domains is selected from the group consisting of SEQ ID NO:1 and SEQ ID NO:2. Or it will be.

[0036] One embodiment of the present invention is a method for the preparation of 180 mg / mL ARGX-113 in 20 mM histidine / histidine HCl, 60 mM sucrose, sucrose, 100 mM arginine HCl, 10 mM L-methionine, and 0.03% (w / v) polysorbate 20, and an aqueous formulation comprising, at pH 6.0, an isolated antibody comprising a variant Fc region. and an FcRn antagonist, wherein the variant Fc region forms a homodimer. and two Fc domains, wherein the amino acid sequence of each of the Fc domains is selected from the group consisting of SEQ ID NO:1 and SEQ ID NO:2. Or it will be.

[0037] One embodiment of the present invention is a method for the preparation of 200 mg / mL ARGX-113 in 20 mM histidine / histidine HCl, 60 mM sucrose, sucrose, 100 mM arginine HCl, 10 mM L-methionine, and 0.03% (w / v) polysorbate 20, and an aqueous formulation comprising, at pH 6.0, an isolated antibody comprising a variant Fc region. and an FcRn antagonist, wherein the variant Fc region forms a homodimer. and two Fc domains, wherein the amino acid sequence of each of the Fc domains is selected from the group consisting of SEQ ID NO:1 and SEQ ID NO:2. Or it will be.

[0038] One aspect of the invention is to provide a method for treating a patient with ARGX-113 at 250 mg / mL, 300 mg / mL, or greater than 300 mg / mL at a pH of about 6.0. 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM arginine HCl, 10 mM L -methionine, and 0.03% (w / v) polysorbate 20, -113 is an isolated FcRn antagonist consisting of a variant Fc region, The mutant Fc region consists of two Fc domains that form a homodimer, where the Fc domains The amino acid sequence of each of the ins consists of SEQ ID NO:1.

[0039] One aspect of the present invention is an isolated neonatal Fc receptor (FcRn) antagonist at about 100 to 300 mg / mL. The compound was dissolved in about 50 mM histidine / histidine HCl, about 200 mM arginine HCl, having a pH of 6.5. wherein the isolated FcRn antagonist is an aqueous formulation comprising a variant Fc region. wherein the variant Fc region comprises two Fc domains that form a homodimer. wherein the amino acid sequence of each of the Fc domains is SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO: It consists of No. 3.

[0040] In one embodiment, the aqueous formulation contains about 200-300 mg / mL of isolated neonatal Fc receptor (FcRn ) The antagonist was dissolved in 50 mM histidine / histidine HCl, 200 mM arginine, with a pH of 6.5. and in HCl, wherein the isolated FcRn antagonist comprises a variant Fc region. wherein the variant Fc region consists of two Fc domains that form a homodimer; Here, the amino acid sequence of each of the Fc domains consists of SEQ ID NO:1.

[0041] In one embodiment, the aqueous formulation contains about 250-300 mg / mL of isolated neonatal Fc receptor (FcRn ) The antagonist was dissolved in 50 mM histidine / histidine HCl, 200 mM arginine, with a pH of 6.5. and in HCl, wherein the isolated FcRn antagonist comprises a variant Fc region. wherein the variant Fc region consists of two Fc domains that form a homodimer; Here, the amino acid sequence of each of the Fc domains consists of SEQ ID NO:1.

[0042] In certain embodiments of each of the aforementioned aspects of the invention, the aqueous formulation is substantially free of dissolved oxygen. Does not include.

[0043] In certain embodiments of each of the aforementioned aspects of the invention, the aqueous formulation is suitable for in vivo use. This is preferable.

[0044] In certain embodiments of each of the aforementioned aspects of the invention, the aqueous formulation is for subcutaneous use in vivo. It is suitable for use.

[0045] One aspect of the invention is to provide a method for the preparation of a pharmaceutical composition comprising administering to a patient a therapeutically effective amount of the aqueous formulation of any one of the preceding aspects and embodiments. A packaged pharmaceutical product comprising a sterile container containing the

[0046] One aspect of the invention is to provide a method for the preparation of a pharmaceutical composition comprising administering to a patient a therapeutically effective amount of the aqueous formulation of any one of the preceding aspects and embodiments. The device includes:

[0047] In some embodiments, the device includes or dispenses from a syringe containing the aqueous formulation. become.

[0048] In some embodiments, the syringe is a pre-filled syringe. [Brief description of the drawings]

[0049] BRIEF DESCRIPTION OF THE DRAWINGS [Figure 1] FIG. 1 shows the shear thinning / thickening behavior of ARGX-113 at the maximum concentration tested with a ramped shear rate from 0 to 4000 s-1. [Diagram 2] Figure 2 shows the concentration (mg / mL) versus viscosity (mPa·s) of ARGX-113 in HisHCl+ salt at the indicated concentrations and temperatures (°C). [Diagram 3] Figure 3 shows protein concentrations measured by UV / Vis for ARGX-113 formulations described in Example 2. sh5, shaking at 5°C; sh25, shaking at 25°C; 4w5, 4 weeks at 5°C; 4w25, 4 weeks at 25°C; 4w40, 4 weeks at 40°C; 8w5, 8 weeks at 5°C; 8w25, 8 weeks at 25°C; 4w40, 8 weeks at 40°C. [Figure 4]4A and 4B respectively show the results of size exclusion chromatography (SEC) of the main peak and high molecular weight (HMW) species of the ARGX-113 formulation described in Example 2. sh5, shaking at 5°C; sh25, shaking at 25°C; 4w5, 4 weeks at 5°C; 4w25, 4 weeks at 25°C; 4w40, 4 weeks at 40°C; 8w5, 8 weeks at 5°C; 8w25, 8 weeks at 25°C; 4w40, 8 weeks at 40°C. [Diagram 5] 5A-5C show the chemical degradation determined by iCE of the main peak, acidic variant, and basic variant of the ARGX-113 formulation described in Example 2, respectively: sh5, shaking at 5° C.; sh25, shaking at 25° C.; 4w5, 4 weeks at 5° C.; 4w25, 4 weeks at 25° C.; 4w40, 4 weeks at 40° C.; 8w5, 8 weeks at 5° C.; 8w25, 8 weeks at 25° C.; 4w40, 8 weeks at 40° C. [Figure 6] 6A-6D show non-visible particles of diameters ≧2 μm, ≧5 μm, ≧10 μm, and ≧25 μm, respectively, for the ARGX-113 formulation described in Example 2. sh5, shaking at 5° C.; sh25, shaking at 25° C.; 4w5, 4 weeks at 5° C.; 4w25, 4 weeks at 25° C.; 4w40, 4 weeks at 40° C.; 8w5, 8 weeks at 5° C.; 8w25, 8 weeks at 25° C.; 4w40, 8 weeks at 40° C. [Figure 7] FIG. 7 shows the turbidity of the ARGX-113 formulations described in Example 2. [Figure 8] Figure 8 shows protein concentration as measured by UV / Vis for ARGX-113 formulations described in Example 3. sh5, shaking at 5°C; sh25, shaking at 25°C; 2w5, 2 weeks at 5°C; 2w25, 2 weeks at 25°C; 2w40, weeks at 40°C. [Figure 9] FIG. 9 shows the osmolality as measured by freezing point depression of the ARGX-113 formulations described in Example 3. [Figure 10]Figure 10 shows the turbidity of the ARGX-113 formulations described in Example 3: sh 5, shake at 5°C; sh 25, shake at 25°C; 2w5, 2 weeks at 5°C; 2w25, 2 weeks at 25°C; 2w40, weeks at 40°C. [Figure 11] 11A and 11B respectively show the results of size exclusion chromatography (SEC) of the main peak and high molecular weight (HMW) species of the ARGX-113 formulation described in Example 3. sh 5, shaking at 5°C; sh 25, shaking at 25°C; 2w5, 2 weeks at 5°C; 2w25, 2 weeks at 25°C; 2w40, weeks at 40°C. [Figure 12] 12A-12C show the chemical degradation determined by iCE of the main peak, acidic variant, and basic variant of the ARGX-113 formulation described in Example 3, respectively: sh5, shaking at 5° C.; sh25, shaking at 25° C.; 2w5, 2 weeks at 5° C.; 2w25, 2 weeks at 25° C.; 2w40, weeks at 40° C. [Figure 13] 13A-13D show subvisible particles of diameters ≧2 μm, ≧5 μm, ≧10 μm, and ≧25 μm, respectively, for the ARGX-113 formulation described in Example 3. sh 5, shaking at 5° C.; sh 25, shaking at 25° C.; 2w5, 2 weeks at 5° C.; 2w25, 2 weeks at 25° C.; 2w40, weeks at 40° C. [Figure 14] Figure 14 shows the turbidity of the ARGX-113 formulations described in Example 4: sh 5, shake at 5°C; sh 25, shake at 25°C; 3w5, 3 weeks at 5°C; 3w25, 3 weeks at 25°C; 3w40, 3 weeks at 40°C; 6w5, 6 weeks at 5°C; 6w25, ​​6 weeks at 25°C; 6w40, 6 weeks at 40°C; 9w5, 9 weeks at 5°C; 9w25, 9 weeks at 25°C; 9w40, 9 weeks at 40°C. [Figure 15]15A and 15B respectively show the results of size exclusion chromatography (SEC) of the main peak and high molecular weight (HMW) species of the ARGX-113 formulation described in Example 4. sh 5, shaking at 5°C; sh 25, shaking at 25°C; 3w5, 3 weeks at 5°C; 3w25, 3 weeks at 25°C; 3w40, 3 weeks at 40°C; 6w5, 6 weeks at 5°C; 6w25, ​​6 weeks at 25°C; 6w40, 6 weeks at 40°C; 9w5, 9 weeks at 5°C; 9w25, 9 weeks at 25°C; 9w40, 9 weeks at 40°C. [Figure 16] 16A-16C show the chemical degradation determined by iCE of the main peak, acidic variant, and basic variant of the ARGX-113 formulation described in Example 4, respectively: sh 5, shaking at 5°C; sh 25, shaking at 25°C; 3w5, 3 weeks at 5°C; 3w25, 3 weeks at 25°C; 3w40, 3 weeks at 40°C; 6w5, 6 weeks at 5°C; 6w25, ​​6 weeks at 25°C; 6w40, 6 weeks at 40°C; 9w5, 9 weeks at 5°C; 9w25, 9 weeks at 25°C; 9w40, 9 weeks at 40°C. [Figure 17] 17A-17D show non-visible particles of diameters ≧2 μm, ≧5 μm, ≧10 μm, and ≧25 μm, respectively, for the ARGX-113 formulation described in Example 4. sh 5, shaking at 5° C.; sh 25, shaking at 25° C.; 3w5, 3 weeks at 5° C.; 3w25, 3 weeks at 25° C.; 3w40, 3 weeks at 40° C.; 6w5, 6 weeks at 5° C.; 6w25, ​​6 weeks at 25° C.; 6w40, 6 weeks at 40° C.; 9w5, 9 weeks at 5° C.; 9w25, 9 weeks at 25° C.; 9w40, 9 weeks at 40° C. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0050] (Detailed Description) (ARGX-113) In certain embodiments, the isolated FcRn antagonist comprises a variant Fc region. wherein the variant Fc region is composed of two Fc domains that form a homodimer, wherein the amino acid sequence of each of the Fc domains is selected from SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3. ARGX-113 is a variant Fc region of human IgG1, wherein the Fc region comprises They contain the amino acids Y, T, E, K, F, and Y at EU positions 252, 254, 256, 433, 434, and 436, respectively. nothing.

[0051] In particular, ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc region, wherein the variant Fc region consists of an Fc domain homodimer, wherein the Fc domain The amino acid sequence of each of these consists of SEQ ID NO:1. [ka]

[0052] The N-terminal aspartic acid residue (D) of SEQ ID NO:1 corresponds to EU position 221 and the C-terminal Lysine (K) corresponds to EU position 447.

[0053] In some embodiments, the aqueous formulations and pharmaceutical compositions of the invention contain a polypeptide of SEQ ID NO:1. In some embodiments, the aqueous formulations and pharmaceutical compositions of the present invention are substantially homogeneous. The article includes a population of polypeptides, wherein at least 90% of the polypeptides (e.g., At least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more More preferably, the Fc domain homodimer comprises The amino acid sequence consists of SEQ ID NO:1.

[0054] In one embodiment, the aqueous formulations and pharmaceutical compositions of the invention contain the polypeptide of SEQ ID NO:2. In certain preferred embodiments, the aqueous formulation of the present invention is substantially homogeneous with respect to the amount of the tide. and a pharmaceutical composition comprising a population of polypeptides, wherein at least 90% of the polypeptides are % (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more) comprises an Fc domain homodimer, wherein the Fc domain of the homodimer The amino acid sequence of each of the main polypeptides consists of SEQ ID NO:2. More than 90% of the peptides lack the C-terminal lysine residue (K) at EU position 448.

[0055] In one embodiment, each Fc domain of ARGX-113 comprises an N-linked glycan at EU position 297. wherein the N-linked glycan is a bisecting N-acetylglucosamine. It has a methylamino(GlcNAc) structure.

[0056] In some embodiments, the aqueous formulations and pharmaceutical compositions of the invention contain a polypeptide of SEQ ID NO:3. In some embodiments, the aqueous formulations and pharmaceutical compositions of the present invention are substantially homogeneous. The article includes a population of polypeptides, wherein at least 90% of the polypeptides (e.g., At least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more More preferably, the Fc domain homodimer comprises The amino acid sequence consists of SEQ ID NO:3.

[0057] (Antibody-mediated autoimmune disease) The formulations and compositions of the present invention are useful in the treatment of antibody-mediated and / or antibody-associated autoimmune diseases. Many applications will be found in this area.

[0058] Antibody-mediated and / or antibody-associated autoimmune diseases are well known. Non-limiting examples of associated autoimmune diseases include pancreatic islet transplant rejection, alopecia areata, and ankylosing leukemia. Spondylitis, antiphospholipid syndrome, autoimmune Addison's disease, Alzheimer's disease, antineutrophil cytoplasmic anti Autoimmune diseases of the adrenal gland (ANCA), autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune myopathy inflammation, autoimmune neutropenia, autoimmune oophoritis and orchitis, autoimmune thrombocytopenia, Autoimmune urticaria, Behcet's disease, bullous pemphigoid, cardiomyopathy, Castleman syndrome, Celiac sprue dermatitis, chronic fatigue immune dysfunction syndrome, chronic inflammatory demyelinating polyneuropathy CIDP, Churg-Strauss syndrome, cicatricial pemphigoid, CREST (calcinosis, Raynaud's phenomenon) , esophageal motility disorder, digital sclerosis, and telangiectasia syndrome, cold agglutinin disease, Crohn's disease , dermatomyositis, discoid lupus, mixed essential cryoglobulinemia, factor VIII deficiency, fibromyalgia Myalgia - Fibromyositis, Glomerulonephritis, Graves' disease, Guillain-Barre syndrome, Goodpasture's syndrome syndrome, graft-versus-host disease (GVHD), Hashimoto's thyroiditis, hemophilia A, idiopathic pulmonary fibrosis, idiopathic hematologic malignancies Thrombocytopenic purpura (immune thrombocytopenia; ITP), IgA neuropathy, IgM polyneuropathy , immune-mediated thrombocytopenia, juvenile arthritis, Kawasaki disease, lichen planus, lupus erythematosus, Nierre's disease, mixed connective tissue disease, multiple sclerosis, type 1 diabetes, multifocal motor neuropathy - (MMN), myasthenia gravis (MG), paraneoplastic bullous pemphigoid, pemphigus vulgaris, pemphigus foliaceus, Pernicious anemia, polyarteritis nodosa, polychondritis, polyglandular syndrome, polymyalgia rheumatica, polymyalgia Myositis and dermatomyositis, primary agammaglobulinemia, primary biliary cirrhosis, psoriasis, psoriatic Arthritis, Raynaud's phenomenon, Reiter's syndrome, rheumatoid arthritis, sarcoidosis, scleroderma, Egren's syndrome, solid organ graft rejection, stiff-man syndrome, systemic lupus erythematosus Systemic lupus erythematosus (SLE), Takayasu's arteritis, toxic epidermal necrolysis (TEN), Stevens-Johnson syndrome (S JS), temporal arteritis / giant cell arteritis, thrombotic thrombocytopenic purpura, ulcerative colitis, uveal ulcer vitiligo, dermatitis herpetiformis vasculitis, antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis, vitiligo, and Wegener's disease Granulomatous disease is one example.

[0059] In certain embodiments, the antibody-mediated autoimmune disease is immune thrombocytopenia (ITP) and severe thrombocytopenia. Myasthenia gravis (MG).

[0060] (formulation) The formulations and compositions of the invention are intended to reduce serum levels of an Fc-containing drug in a subject. Fc-containing drugs may find use in any disease or disorder for which the drug is desired. These include, but are not limited to, autoantibodies, therapeutic antibodies, diagnostic antibodies, and immune complexes. Further non-limiting examples of Fc-containing agents include imaging agents (e.g., labeled antibodies), antibodies -drug conjugates (ADCs), Fc fusion proteins (e.g., immunoadhesins), and immunogens (e.g., non-human antibodies).

[0061] Furthermore, in a disease or illness requiring administration of a therapeutic agent, the subject may develop resistance to the therapeutic agent. These often result in the production of anti-drug antibodies (e.g., anti-drug antibodies) that, in turn, inhibit the therapeutic agent from acting as intended. or prevent it from being used for the intended therapeutic purpose or cause adverse reactions in the subject. Thus, the formulations and compositions disclosed herein are intended to prevent or reduce the therapeutic effects that may occur in a subject. It may also be used to remove antibodies against the therapeutic agent (eg, anti-drug antibodies).

[0062] One aspect of the present invention is an isolated neonatal Fc receptor (FcRn) antagonist at about 100 to 300 mg / mL. The mixture was diluted with 20-60 mM histidine / histidine HCl, 0-70 mM sucrose, 0-150 mM NaCl, 0-250 mM mM arginine HCl, 0% to 0.05% (w / v) polysorbate 20 or polysorbate 80, 0 to 15 mM L -methionine, pH 6.0 to 6.5, wherein the isolated FcRn antagonist is The antigen comprises a variant Fc region, wherein the variant Fc region forms a homodimer. and wherein the amino acid sequence of each of the Fc domains is the sequence It consists of sequence number 1, sequence number 2, or sequence number 3.

[0063] In one embodiment, the aqueous formulation comprises about 100-300 mg / mL of isolated neonatal Fc receptor (Fc Rn) antagonists were diluted in 20-60 mM histidine / histidine HCl, 0-70 mM sucrose, 100 mM NaCl, 0% to 0.05% (w / v) polysorbate 20 or polysorbate 80, 0 to 15 mM L-methionine and pH 6.0 to 6.5, wherein the isolated FcRn antagonist comprises a variant Fc domain. wherein the variant Fc region is composed of two Fc domains that form a homodimer. wherein the amino acid sequence of each of the Fc domains is selected from SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3. It consists of column number 3.

[0064] In one embodiment, the aqueous formulation comprises about 100-300 mg / mL of isolated neonatal Fc receptor (Fc Rn) antagonists were added to 20-60 mM histidine / histidine HCl, 0-70 mM sucrose, 100-2 50 mM Arginine HCl, 0% to 0.05% (w / v) Polysorbate 20 or Polysorbate 80, 0 to 15 mM L-methionine, pH 6.0 to 6.5, wherein the isolated FcRn antagonist is wherein the variant Fc region comprises two F domains that form a homodimer. and c domains, wherein the amino acid sequence of each of the Fc domains is SEQ ID NO:1, SEQ ID NO:2, No. 2, or No. 3.

[0065] In one embodiment, the aqueous formulation comprises about 100-300 mg / mL of isolated neonatal Fc receptor (Fc Rn) antagonists were selected from 20-60 mM histidine / histidine HCl, 100-250 mM arginine HCl, 0%-0.05% (w / v) polysorbate 20 or polysorbate 80, 0-15 mM L-methionine, pH 6 0.0 to 6.5, wherein the isolated FcRn antagonist comprises a variant Fc region. wherein the variant Fc region consists of two Fc domains that form a homodimer. wherein the amino acid sequence of each of the Fc domains is SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO: It consists of 3.

[0066] In each of the above aspects and embodiments, in certain embodiments, the aqueous formulation comprises 20 or Contains 50mM histidine.

[0067] In each of the above aspects and embodiments, in certain embodiments, the aqueous formulation comprises 0.02% Contains ~0.04% polysorbate 20 or polysorbate 80.

[0068] In each of the above aspects and embodiments, in certain embodiments, the aqueous formulation comprises 0 or 1 Contains 0 mM L-methionine.

[0069] In each of the above aspects and embodiments, in certain embodiments, the pH is 6.0 or 6.5. be.

[0070] In each of the foregoing aspects and embodiments, in certain embodiments, each of the Fc domains The amino acid sequence consists of SEQ ID NO:1.

[0071] In one embodiment of the present invention, an isolated FcRn antagonist is dissolved in about 100 to about 300 mg / mL of 20 mM histidine. The solution consisted of 10 mM lysine / histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.02% to 0.04% (w / v) polyisobutene. and polysorbate 20 or polysorbate 80, pH 6.0, wherein the isolated The FcRn antagonist comprises a variant Fc region, wherein the variant Fc region comprises: It consists of two Fc domains that form a homodimer, where each amino acid of the Fc domain is The acid sequence consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0072] In one embodiment of the present invention, an isolated FcRn antagonist is dissolved in about 100 to about 200 mg / mL of 20 mM histidine. The solution consisted of 10 mM lysine / histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.02% to 0.04% (w / v) polyisobutene. and polysorbate 20 or polysorbate 80, pH 6.0, wherein the isolated The FcRn antagonist comprises a variant Fc region, wherein the variant Fc region comprises: It consists of two Fc domains that form a homodimer, where each amino acid of the Fc domain is The acid sequence consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0073] One aspect of the present invention is an isolated neonatal Fc receptor (FcRn) antagonist at about 100 to 300 mg / mL. The samples were prepared in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0% to 0.04% ( (w / v) polysorbate 20 or polysorbate 80, pH 6.0, wherein The isolated FcRn antagonist comprises a variant Fc region, wherein the variant The Fc region consists of two Fc domains that form a homodimer, where each of the Fc domains Each amino acid sequence consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0074] In one embodiment, the present invention relates to a method for the preparation of a medicament for the treatment of atopic dermatitis with a subject comprising administering about 100 to about 300 mg / mL of ARGX-113 in 20 mM histidine / Histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.02% to 0.04% (w / v) polysorbate 20 or polysorbate 80, pH 6.0, wherein ARGX-113 is an aqueous formulation containing an isolated FcRn antagonist comprising a variant Fc region, It consists of two Fc domains that form a homodimer, where each agonist of the Fc domain The amino acid sequence consists of SEQ ID NO:1.

[0075] One aspect of the present invention is an isolated neonatal Fc receptor (FcRn) antagonist at about 100 to 200 mg / mL. The solution was prepared in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.02% to 0.04% ethanol. % (w / v) polysorbate 20 or polysorbate 80, pH 6.0, The isolated FcRn antagonist comprises a variant Fc region, wherein the variant The Fc region of a serogroup is composed of two Fc domains that form a homodimer, where the Fc domains Each of the amino acid sequences consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0076] In one embodiment, the present invention relates to a method for the preparation of a pharmaceutical composition comprising administering about 100 to about 200 mg / mL of ARGX-113 in 20 mM histidine / Histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.02% to 0.04% (w / v) polysorbate 20 or polysorbate 80, pH 6.0, wherein ARGX-113 is an aqueous formulation containing an isolated FcRn antagonist comprising a variant Fc region, It consists of two Fc domains that form a homodimer, where each agonist of the Fc domain The amino acid sequence consists of SEQ ID NO:1.

[0077] As used herein, the term "about" means ±10% of any given specified amount. For example, about 200 mg / mL means 90% to 110% of 200 mg / mL, that is, 180 to 220 mg. g / mL.

[0078] In some embodiments, the aqueous formulation comprises 100-300 mg / mL of ARGX-113.

[0079] In some embodiments, the aqueous formulation comprises 100-200 mg / mL of ARGX-113.

[0080] In some embodiments, the aqueous formulation contains about 150 to about 200 mg / mL of ARGX-113. In an embodiment, the aqueous formulation comprises 150-200 mg / mL of ARGX-113.

[0081] In some embodiments, the aqueous formulation contains about 150 to about 180 mg / mL of ARGX-113. In an embodiment, the aqueous formulation comprises 150-180 mg / mL of ARGX-113.

[0082] In some embodiments, the aqueous formulation comprises about 165 mg / mL of ARGX-113.

[0083] In some embodiments, the aqueous formulation comprises about 175 mg / mL of ARGX-113.

[0084] In some embodiments, the aqueous formulation comprises about 180 mg / mL of ARGX-113.

[0085] In some embodiments, the aqueous formulation comprises about 250 mg / mL of ARGX-113.

[0086] In some embodiments, the aqueous formulation comprises about 300 mg / mL of ARGX-113.

[0087] According to each of the above embodiments, in some embodiments, the aqueous formulation comprises 0.02% to 0.04% (w / v) polysorbate 20. In some embodiments, the aqueous formulation contains 0.02% (w / v) polysorbate 20. In one embodiment, the aqueous formulation contains 0.03% (w / v) polysorbate 20. In some embodiments, the aqueous formulation comprises 0.04% (w / v) polysorbate 20.

[0088] Alternatively, according to each of the preceding embodiments, in some embodiments, the aqueous formulation comprises 0.02% ~0.04% (w / v) polysorbate 80. In some embodiments, the aqueous formulation contains 0.02% (w / v) Polysorbate 80. In certain embodiments, the aqueous formulation contains 0.03% (w / v) Polysorbate 80. In one embodiment, the aqueous formulation contains 0.04% (w / v) polysorbate 80. include.

[0089] In one embodiment, the aqueous formulation contains 165 mg / mL ARGX-113 in 20 mM histidine / histidine. HCl, 60 mM sucrose, 100 mM NaCl, and 0.04% (w / v) polysorbate 20, pH 6.0. ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc region. wherein the variant Fc region consists of two Fc domains that form a homodimer; Here, the amino acid sequence of each of the Fc domains consists of SEQ ID NO:1.

[0090] One embodiment of the present invention is a method for the preparation of an isolated FcRn antagonist in a suspension of about 150 mg / mL of 20 mM histidine / Histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.04% (w / v) polysorbate 20, pH 6 0.0, wherein the isolated FcRn antagonist is a variant FcRn antagonist. wherein the variant Fc region comprises two Fc domains that form a homodimer. wherein the amino acid sequence of each of the Fc domains is SEQ ID NO:1, SEQ ID NO:2, or It consists of sequence number 3.

[0091] In one embodiment, the present invention provides a method for treating ARGX-113 at 150 mg / mL in 20 mM histidine / histidine H Cl, 60 mM sucrose, 100 mM NaCl, and 0.04% (w / v) polysorbate 20, pH 6.0 ARGX-113 is an aqueous formulation, wherein the ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc region. agonist, wherein the variant Fc region comprises two Fc domains that form a homodimer. wherein the amino acid sequence of each of the Fc domains consists of SEQ ID NO:1.

[0092] One embodiment of the present invention is a method for the preparation of an isolated FcRn antagonist in a suspension of about 175 mg / mL in 20 mM histidine / Histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.04% (w / v) polysorbate 20, pH 6 0.0, wherein the isolated FcRn antagonist is a variant FcRn antagonist. wherein the variant Fc region comprises two Fc domains that form a homodimer. wherein the amino acid sequence of each of the Fc domains is SEQ ID NO:1, SEQ ID NO:2, or It consists of sequence number 3.

[0093] In one embodiment, the present invention provides a method for treating ARGX-113 at 175 mg / mL in 20 mM histidine / histidine H Cl, 60 mM sucrose, 100 mM NaCl, and 0.04% (w / v) polysorbate 20, pH 6.0 ARGX-113 is an aqueous formulation, wherein the ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc region. agonist, wherein the variant Fc region comprises two Fc domains that form a homodimer. wherein the amino acid sequence of each of the Fc domains consists of SEQ ID NO:1.

[0094] One embodiment of the present invention is a method for the preparation of an isolated FcRn antagonist in a concentration of about 200 mg / mL in 20 mM histidine / Histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.04% (w / v) polysorbate 20, pH 6 0.0, wherein the isolated FcRn antagonist is a variant FcRn antagonist. wherein the variant Fc region comprises two Fc domains that form a homodimer. wherein the amino acid sequence of each of the Fc domains is SEQ ID NO:1, SEQ ID NO:2, or It consists of sequence number 3.

[0095] In one embodiment, the present invention provides a method for treating ARGX-113 at 200 mg / mL in 20 mM histidine / histidine H Cl, 60 mM sucrose, 100 mM NaCl, and 0.04% (w / v) polysorbate 20, pH 6.0 ARGX-113 is an aqueous formulation, wherein the ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc region. agonist, wherein the variant Fc region comprises two Fc domains that form a homodimer. wherein the amino acid sequence of each of the Fc domains consists of SEQ ID NO:1.

[0096] In one embodiment of the present invention, about 100 to 300 mg / mL of an isolated FcRn antagonist is dissolved in 20 mM histidine. dl / histidine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0% to 0.04% % (w / v) polysorbate 20 or polysorbate 80, pH 6.0, The isolated FcRn antagonist comprises a variant Fc region, wherein the variant The Fc region of a serogroup is composed of two Fc domains that form a homodimer, where the Fc domains Each of the amino acid sequences consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0097] In one embodiment, the present invention relates to a method for treating a patient having an ARGX-113 deficiency, comprising administering about 100 to 300 mg / mL of ARGX-113 in a 20 mM histidine / histidine / sodium glycerol solution. thionine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0% to 0.04% (w / v) polysaccharide. An aqueous formulation comprising resorbate 20 or polysorbate 80, pH 6.0, wherein ARGX-1 13 is an isolated FcRn antagonist consisting of a variant Fc region, The ant Fc region consists of two Fc domains that form a homodimer, where the Fc domains The amino acid sequence of each of the peptides consists of SEQ ID NO:1.

[0098] In one embodiment of the present invention, about 100 to 200 mg / mL of an isolated FcRn antagonist is dissolved in 20 mM histidine. ds / histidine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.02% An aqueous formulation containing 0.04% (w / v) polysorbate 20 or polysorbate 80, pH 6.0, wherein the isolated FcRn antagonist comprises a variant Fc region, The ant Fc region consists of two Fc domains that form a homodimer, where the Fc domains The amino acid sequence of each of the peptides consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0099] In one embodiment, the present invention relates to a method for treating a patient having an ARGX-113 deficiency of about 100-200 mg / mL in a 20 mM histidine / histidine / sodium phosphate buffer. Thymine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.02% to 0.04% (w / v ) Polysorbate 20 or Polysorbate 80, an aqueous formulation containing ARG in pH 6.0 X-113 is an isolated FcRn antagonist consisting of a variant Fc region, wherein the The variant Fc region consists of two Fc domains that form a homodimer, where the Fc domains The main amino acid sequence of each consists of SEQ ID NO:1.

[0100] One embodiment of the invention provides an isolated neonatal Fc receptor (FcRn) antagonist at about 165 mg / mL in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and and 0.04% (w / v) polysorbate 20, pH 6.0, wherein the isolated The FcRn antagonist comprises a variant Fc region, wherein the variant Fc region comprises: It consists of two Fc domains that form a homodimer, where each amino acid of the Fc domain is The acid sequence consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0101] In one embodiment, the present invention provides a method for treating ARGX-113 at 165 mg / mL in 20 mM histidine / histidine H Cl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.04% (w / v) polysorbate. 20, pH 6.0, wherein ARGX-113 is a single polypeptide comprising a variant Fc region. an isolated FcRn antagonist, wherein the variant Fc region forms a homodimer. and two Fc domains corresponding to each other, wherein the amino acid sequence of each of the Fc domains is It consists of No. 1.

[0102] One embodiment of the present invention is a method for the preparation of an isolated FcRn antagonist in a suspension of about 175 mg / mL in 20 mM histidine / Histidine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.03% (w / v) polysaccharide. resorbate 20, pH 6.0, wherein the isolated FcRn antagonist is The antibody comprises a variant Fc region, wherein the variant Fc region forms a homodimer. and two Fc domains corresponding to each other, wherein the amino acid sequence of each of the Fc domains is SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.

[0103] In one embodiment, the present invention provides a method for treating ARGX-113 at 175 mg / mL in 20 mM histidine / histidine H Cl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.03% (w / v) polysorbate. 20, pH 6.0, wherein ARGX-113 is a single polypeptide comprising a variant Fc region. an isolated FcRn antagonist, wherein the variant Fc region forms a homodimer. and two Fc domains corresponding to each other, wherein the amino acid sequence of each of the Fc domains is It consists of No. 1.

[0104] One embodiment of the present invention is a method for the preparation of an isolated FcRn antagonist in a concentration of about 200 mg / mL in 20 mM histidine / Histidine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.03% (w / v) polysaccharide. resorbate 20, pH 6.0, wherein the isolated FcRn antagonist is The antibody comprises a variant Fc region, wherein the variant Fc region forms a homodimer. and two Fc domains corresponding to each other, wherein the amino acid sequence of each of the Fc domains is SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.

[0105] In one embodiment, the present invention provides a method for treating ARGX-113 at 200 mg / mL in 20 mM histidine / histidine H Cl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.03% (w / v) polysorbate. 20, pH 6.0, wherein ARGX-113 is a single polypeptide comprising a variant Fc region. and an isolated FcRn antagonist, wherein the variant Fc region is an Fc domain homodimer. wherein the amino acid sequence of each of the Fc domains consists of SEQ ID NO:1.

[0106] In one embodiment of the present invention, an isolated FcRn antagonist is dissolved in about 100 to about 300 mg / mL of 50 mM histidine. Thysidine / histidine HCl, 60 mM sucrose, 150 mM arginine HCl, and 0.02% to 0.04% (w / v ) an aqueous formulation containing polysorbate 20 or polysorbate 80, pH 6.0, wherein The isolated FcRn antagonist comprises a variant Fc region, wherein the variant Fc The region consists of two Fc domains that form a homodimer, where each of the Fc domains The amino acid sequence consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0107] In one embodiment, the present invention relates to a method for the preparation of a medicament for the treatment of atopic dermatitis with a subject having ... Stidine HCl, 60 mM sucrose, 150 mM arginine HCl, and 0.02% to 0.04% (w / v) Polysol and an aqueous formulation comprising ARGX-113 in polysorbate 20 or polysorbate 80, pH 6.0, An isolated FcRn antagonist comprising a variant Fc region, wherein the variant The Fc region consists of two Fc domains that form a homodimer, where each of the Fc domains Each amino acid sequence consists of SEQ ID NO:1.

[0108] In one embodiment according to this aspect of the invention, the aqueous formulation contains 100-300 mg / mL of ARGX-11 Includes 3.

[0109] In one embodiment according to this aspect of the invention, the aqueous formulation contains about 150 to about 200 mg / mL of ARG. In one embodiment, according to this aspect of the invention, the aqueous formulation comprises 150 to 20 Contains 0 mg / mL ARGX-113.

[0110] In one embodiment according to this aspect of the invention, the aqueous formulation contains about 150 to about 180 mg / mL of ARG. In one embodiment, according to this aspect of the invention, the aqueous formulation comprises 150-180 Contains 100 mg / mL of ARGX-113.

[0111] According to each of the above embodiments, in some embodiments, the aqueous formulation comprises 0.02% to 0.04% (w / v) polysorbate 20. In some embodiments, the aqueous formulation contains 0.02% (w / v) polysorbate 20. In one embodiment, the aqueous formulation contains 0.03% (w / v) polysorbate 20. In some embodiments, the aqueous formulation comprises 0.04% (w / v) polysorbate 20.

[0112] In one embodiment of the present invention, an isolated FcRn antagonist is dissolved in about 100 to about 200 mg / mL of 50 mM histidine. Thysidine / histidine HCl, 60 mM sucrose, 150 mM arginine HCl, and 0.02% to 0.04% (w / v ) an aqueous formulation containing polysorbate 20 or polysorbate 80, pH 6.0, wherein The isolated FcRn antagonist comprises a variant Fc region, wherein the variant Fc The region consists of two Fc domains that form a homodimer, where each of the Fc domains The amino acid sequence consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0113] In one embodiment, the present invention relates to a method for the preparation of a medicament for the treatment of atopic dermatitis with a subject having ... Stidine HCl, 60 mM sucrose, 150 mM arginine HCl, and 0.02% to 0.04% (w / v) Polysol and an aqueous formulation comprising ARGX-113 in polysorbate 20 or polysorbate 80, pH 6.0, An isolated FcRn antagonist comprising a variant Fc region, wherein the variant The Fc region consists of two Fc domains that form a homodimer, where each of the Fc domains Each amino acid sequence consists of SEQ ID NO:1.

[0114] In one embodiment according to this aspect of the invention, the aqueous formulation contains 100-300 mg / mL of ARGX-11 Includes 3.

[0115] In one embodiment according to this aspect of the invention, the aqueous formulation contains about 150 to about 200 mg / mL of ARG. In one embodiment, according to this aspect of the invention, the aqueous formulation comprises 150 to 20 Contains 0 mg / mL ARGX-113.

[0116] In one embodiment according to this aspect of the invention, the aqueous formulation contains about 150 to about 180 mg / mL of ARG. In one embodiment, according to this aspect of the invention, the aqueous formulation comprises 150 to 18 Contains 0 mg / mL ARGX-113.

[0117] According to each of the above embodiments, in some embodiments, the aqueous formulation comprises 0.02% to 0.04% (w / v) polysorbate 20. In some embodiments, the aqueous formulation contains 0.02% (w / v) polysorbate 20. In one embodiment, the aqueous formulation contains 0.03% (w / v) polysorbate 20. In some embodiments, the aqueous formulation comprises 0.04% (w / v) polysorbate 20.

[0118] Alternatively, according to each of the preceding embodiments, in some embodiments, the aqueous formulation comprises 0.02% ~0.04% (w / v) polysorbate 80. In some embodiments, the aqueous formulation contains 0.02% (w / v) Polysorbate 80. In certain embodiments, the aqueous formulation contains 0.03% (w / v) Polysorbate 80. In one embodiment, the aqueous formulation contains 0.04% (w / v) polysorbate 80. include.

[0119] In one embodiment, the aqueous formulation contains about 150 mg / mL of ARGX-113 in 50 mM histidine / histidine. arginine HCl, 60 mM sucrose, 150 mM arginine HCl, and 0.04% (w / v) polysorbate 80, pH Included in 6.0.

[0120] In one embodiment, the aqueous formulation comprises 150 mg / mL ARGX-113 in 50 mM histidine / histidine. HCl, 60 mM sucrose, 150 mM arginine HCl, 10 mM L-methionine, and 0.04% (w / v) polysaccharide. Included in resorbate 80, pH 6.0.

[0121] In one embodiment of the present invention, about 100 to 200 mg / mL of an isolated FcRn antagonist is dissolved in 20 mM histidine. arginine / histidine HCl, 60 mM sucrose, 100 mM arginine HCl, 10 mM L-methionine, and 0 An aqueous formulation containing 0.02% to 0.04% (w / v) polysorbate 20 or polysorbate 80, pH 6.0. wherein the isolated FcRn antagonist consists of a variant Fc region, The variant Fc region is composed of two Fc domains that form a homodimer, wherein the F The amino acid sequence of each of the c domains consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0122] In one embodiment, the present invention relates to a method for treating a patient having an ARGX-113 deficiency of about 100-200 mg / mL in a 20 mM histidine / histidine / sodium phosphate buffer. Thyidine HCl, 60 mM sucrose, 100 mM arginine HCl, 10 mM L-methionine, and 0.02% to 0. It is an aqueous formulation containing 0.4% (w / v) polysorbate 20 or polysorbate 80, pH 6.0, Here, ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc region, wherein the variant Fc region is composed of two Fc domains that form a homodimer, The amino acid sequence of each of the Fc domains consists of SEQ ID NO:1.

[0123] One embodiment of the present invention is a method for the preparation of an isolated FcRn antagonist in a suspension of about 175 mg / mL in 20 mM histidine / Histidine HCl, 60 mM sucrose, 100 mM arginine HCl, 10 mM L-methionine, and 0.03% (w / v) polysorbate 20, pH 6.0, wherein the isolated FcRn antigen is The antagonist comprises a variant Fc region, wherein the variant Fc region is a homodimer. The antibody comprises two Fc domains forming a fusion protein, wherein the amino acid sequence of each of the Fc domains is , SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0124] In one embodiment, the present invention provides a method for treating ARGX-113 at 175 mg / mL in 20 mM histidine / histidine H Cl, 60 mM sucrose, 100 mM arginine HCl, 10 mM L-methionine, and 0.03% (w / v) polyisobutene. The formulation is an aqueous formulation containing ARGX-113 in 1000 mM NaCl, pH 6.0, wherein the ARGX-113 is a variant Fc region. and an isolated FcRn antagonist comprising an FcRn domain comprising: and wherein the amino acid sequence of each of the Fc domains comprises SEQ ID NO:1. do.

[0125] One embodiment of the present invention is a method for the preparation of an isolated FcRn antagonist in a concentration of about 200 mg / mL in 20 mM histidine / Histidine HCl, 60 mM sucrose, 100 mM arginine HCl, 10 mM L-methionine, and 0.03% (w / v) polysorbate 20, pH 6.0, wherein the isolated FcRn antigen is The antagonist comprises a variant Fc region, wherein the variant Fc region comprises an Fc domain wherein the amino acid sequence of each of the Fc domains is SEQ ID NO:1; It consists of sequence number 2 or sequence number 3.

[0126] In one embodiment, the present invention provides a method for treating ARGX-113 at 200 mg / mL in 20 mM histidine / histidine H Cl, 60 mM sucrose, 100 mM arginine HCl, 10 mM L-methionine, and 0.03% (w / v) polyisobutene. The formulation is an aqueous formulation containing ARGX-113 in 1000 mM NaCl, pH 6.0, wherein the ARGX-113 is a variant Fc region. and an isolated FcRn antagonist comprising an FcRn domain comprising: and wherein the amino acid sequence of each of the Fc domains comprises SEQ ID NO:1. do.

[0127] One aspect of the present invention is an isolated neonatal Fc receptor (FcRn) antagonist at about 100 to 300 mg / mL. in an aqueous formulation containing 50 mM histidine / histidine HCl, 200 mM arginine HCl, pH 6.5. wherein the isolated FcRn antagonist consists of a variant Fc region, The variant Fc region is composed of two Fc domains that form a homodimer, wherein the F The amino acid sequence of each of the c domains consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0128] In one embodiment, the aqueous formulation contains about 200-300 mg / mL of isolated neonatal Fc receptor (FcRn ) Antagonists were dissolved in 50 mM histidine / histidine HCl, 200 mM arginine HCl, pH 6.5. wherein the isolated FcRn antagonist is an aqueous formulation comprising a variant Fc region. wherein the variant Fc region comprises two Fc domains that form a homodimer. wherein the amino acid sequence of each of the Fc domains consists of SEQ ID NO:1.

[0129] In one embodiment, the aqueous formulation contains about 250-300 mg / mL of isolated neonatal Fc receptor (FcRn ) Antagonists were dissolved in 50 mM histidine / histidine HCl, 200 mM arginine HCl, pH 6.5. wherein the isolated FcRn antagonist is an aqueous formulation comprising a variant Fc region. wherein the variant Fc region comprises two Fc domains that form a homodimer. wherein the amino acid sequence of each of the Fc domains consists of SEQ ID NO:1.

[0130] According to each of the foregoing aspects and embodiments, in certain embodiments, the aqueous formulation comprises a dissolved acid. As used herein, the term "substantially free" means For example, in some embodiments, the aqueous formulation is at least 95% free of soluble In various embodiments, the aqueous formulation is at least 95% free of dissolved oxygen. Contains at least 96%, at least 97%, at least 98%, at least 99%, or 100% Assuming that the water is equilibrated with an atmosphere that is typically 20% oxygen, in one embodiment In the present invention, an aqueous formulation that is substantially free of dissolved oxygen contains 1% or less dissolved oxygen. In embodiments, the aqueous formulation is 100% free of dissolved oxygen.

[0131] According to each of the above aspects and embodiments, in certain embodiments, the aqueous formulation comprises an in vivo For example, in such an embodiment, the aqueous formulation is sterile and and does not contain medicamentically unacceptable amounts of toxic substances, such as endotoxins. Such aqueous formulations may be formulated, for example, in accordance with regulations issued by the U.S. Food and Drug Administration (FDA). Ability to comply with Good Manufacturing Practice (GMP) quality standards.

[0132] (Method of preparation of formulation) The formulations according to the invention can be prepared using any suitable method. 13 is prepared from a eukaryotic cell containing an expression vector or a nucleic acid sequence encoding the Fc domain. For example, eukaryotic cells include Chinese hamster ovary (CHO) cells, DG44 and DUXB11 (CHO cells). Neesy hamster ovary line, DHFR minus), HELA (human cervical cancer), CVI (monkey kidney line), COS ( CVI derivative with SV40 T antigen), R1610 (Chinese hamster fibroblast), BALBC / 3 T3 (mouse fibroblasts), HAK (hamster kidney line), SP2 / 0 (mouse myeloma), BFA-1c1BPT (human The cells may be human endothelial cells, RAJI (human lymphocytes), 293 (human kidney), or NS0 cells. In one embodiment, the eukaryotic cell used to express ARGX-113 is a CHO cell. See, for example, WO 2015 / 100299, the entire contents of which are incorporated herein by reference. ARGX-113 is typically isolated from cells using techniques known in the art. It is usually expressed as a secreted protein that can be isolated and concentrated. The protein product was then purified by lysis in Tris / glycine, pH 7.2 or 20 mM L-histidine / L-histidine. It can be placed in a sterile aqueous solution such as thiamine HCl·H2O, pH 6.0.

[0133] This initial product is then diluted with ARGX-113 at a concentration equal to or exceeding the target final concentration. If necessary, increase the concentration to reach a concentrated protein solution containing For example, the concentration increase and buffer exchange are carried out by changing about 200 mg / mL of ARGX-113 into 20 mM L-hyd. The intermediate product was produced by diluting the mixture in 100 mM NaCl, pH 6.0 with 100 mM lysine / L-histidine. This can be done.

[0134] Concentration can be accomplished using any suitable method known in the art. Such methods include, but are not limited to, tangential flow filtration (TFF), These include dialysis, ultrafiltration, and freeze-drying. For commercial production, TFF is usually It can be used.

[0135] Additional ingredients can then be added to arrive at the desired final formulation. For example, NaCl Additional ingredients such as arginine HCl, sucrose, and / or polysorbate may be added to the additional ingredients. can be added from concentrated stock solutions of each, and, if desired, water can be added to obtain the desired concentration. In certain embodiments, polysorbate (PS20) can be used to arrive at a final formulation of It is added as the very last excipient in the formulation to ensure that the correct pH value is achieved (Polysorbate). Since the molecular weight of polysorbate is the same as that of ARGX-113, it is recommended to add polysorbate last. (This will avoid an increase in concentration.)

[0136] In some embodiments, the intermediate solution and additional components are degassed or otherwise treated to Reduce or eliminate dissolved oxygen. For example, purify the intermediate solution and additional components with argon or nitrogen. It can be equilibrated.

[0137] In some embodiments, the final aqueous formulation is degassed or otherwise treated to remove dissolved oxygen. For example, the final aqueous formulation may be prepared by reducing or eliminating dissolved oxygen from the formulation. by bubbling argon or nitrogen through the final aqueous formulation for a period of time sufficient to In one embodiment, the final aqueous formulation is then equilibrated with the gas. Store under nitrogen.

[0138] The aqueous formulations so prepared are typically sterile filtered and then subsequently sterilized as described herein. The solution is dispensed and stored in a sterile container or device that has been

[0139] (Route of Administration) The aqueous formulations of the present invention are suitable for parenteral administration. The formulation is suitable for subcutaneous administration. In some embodiments, the aqueous formulation of the present invention is suitable for intravenous administration. In certain embodiments, the aqueous formulations of the present invention are suitable for intraperitoneal administration. .

[0140] (effective dose) The formulations and compositions will generally be administered in an effective amount. In some embodiments, an effective amount refers to a therapeutically effective amount, That is, it refers to an amount sufficient to achieve a desired therapeutic effect in a subject. Examples of such symptoms include, but are not limited to, a decrease in serum total IgG, and the development of myasthenia gravis (MG) and and the treatment of various antibody-mediated autoimmune diseases such as immune thrombocytopenia (ITP).

[0141] (subject) As used herein, a "subject" generally refers to a mammal. In one embodiment, the subject is a mammal other than a human or a non-human primate. The subject is a human or a non-human primate. In some embodiments, the subject is a human. In some embodiments, the subject is an adult, i.e., a human who is at least 18 years of age. In the method, the subject is a human under the age of 18.

[0142] (Pharmaceutical products) One aspect of the present invention is a packaged pharmaceutical composition comprising a sterile container containing a therapeutically effective amount of the aqueous formulation of the present invention. In various embodiments, the packaged pharmaceutical products are single-use vials, multi-use vials, or It can be presented as a multi-use vial or as a pre-filled syringe.

[0143] (Device) One aspect of the invention is a device comprising a therapeutically effective amount of the aqueous formulation of the invention.

[0144] In one embodiment, the aqueous formulation contains about 100-200 mg / mL of an isolated FcRn antagonist. The solution was prepared in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.02% to 0.04% ethanol. % (w / v) polysorbate 20 or polysorbate 80, pH 6.0, The isolated FcRn antagonist comprises a variant Fc region, wherein the variant The Fc region of a serogroup is composed of two Fc domains that form a homodimer, where the Fc domains Each of the amino acid sequences consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0145] In one embodiment, the aqueous formulation contains 150 mg / mL of the isolated FcRn antagonist in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.04% (w / v) Polysol. and an aqueous formulation comprising 100% ethanol at 20° C., pH 6.0, wherein the isolated FcRn antagonist is and a variant Fc region, wherein the variant Fc region is a homodimer-forming two and two Fc domains, wherein the amino acid sequence of each of the Fc domains is set forth in SEQ ID NO:1, It consists of SEQ ID NO:2 or SEQ ID NO:3.

[0146] In one embodiment, the aqueous formulation contains 175 mg / mL of the isolated FcRn antagonist in 20 mL of mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.04% (w / v) Polysol. and an aqueous formulation comprising 100% ethanol at 20° C., pH 6.0, wherein the isolated FcRn antagonist is and a variant Fc region, wherein the variant Fc region is a homodimer-forming two and two Fc domains, wherein the amino acid sequence of each of the Fc domains is set forth in SEQ ID NO:1, It consists of SEQ ID NO:2 or SEQ ID NO:3.

[0147] In one embodiment, the aqueous formulation contains 200 mg / mL of the isolated FcRn antagonist. mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.04% (w / v) Polysol. and an aqueous formulation comprising 100% ethanol at 20° C., pH 6.0, wherein the isolated FcRn antagonist is and a variant Fc region, wherein the variant Fc region is a homodimer-forming two and two Fc domains, wherein the amino acid sequence of each of the Fc domains is set forth in SEQ ID NO:1, It consists of SEQ ID NO:2 or SEQ ID NO:3.

[0148] In one embodiment, the aqueous formulation comprises about 100-300 mg / mL of ARGX-113 in 20 mM histidine / Histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0% to 0.04% (w / v) polysorbate 20 or polysorbate 80, pH 6.0, wherein ARGX-113 is an isolated FcRn antagonist comprising a variant Fc region, wherein the variant Fc region comprises It consists of two Fc domains that form a homodimer, where each amino acid of the Fc domain The amino acid sequence consists of SEQ ID NO:1.

[0149] In one embodiment, the aqueous formulation comprises about 100-200 mg / mL of ARGX-113 in 20 mM histidine / Histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.02% to 0.04% (w / v) polysorbate 20 or polysorbate 80, pH 6.0, wherein ARGX-113 is an aqueous formulation containing an isolated FcRn antagonist comprising a variant Fc region, It consists of two Fc domains that form a homodimer, where each agonist of the Fc domain The amino acid sequence consists of SEQ ID NO:1.

[0150] In one embodiment, the aqueous formulation contains 150 mg / mL ARGX-113 in 20 mM histidine / histidine. in 50 mM NaCl, 60 mM sucrose, 100 mM NaCl, and 0.04% (w / v) polysorbate 20, pH 6.0. wherein ARGX-113 is an aqueous formulation comprising an isolated FcRn antigen comprising a variant Fc region. and wherein the variant Fc region comprises two Fc domains that form a homodimer. wherein the amino acid sequence of each of the Fc domains consists of SEQ ID NO:1.

[0151] In one embodiment, the aqueous formulation contains 175 mg / mL ARGX-113 in 20 mM histidine / histidine. in 50 mM NaCl, 60 mM sucrose, 100 mM NaCl, and 0.04% (w / v) polysorbate 20, pH 6.0. wherein ARGX-113 is an aqueous formulation comprising an isolated FcRn antigen comprising a variant Fc region. and wherein the variant Fc region comprises two Fc domains that form a homodimer. wherein the amino acid sequence of each of the Fc domains consists of SEQ ID NO:1.

[0152] In one embodiment, the aqueous formulation contains 200 mg / mL ARGX-113 in 20 mM histidine / histidine. in 50 mM NaCl, 60 mM sucrose, 100 mM NaCl, and 0.04% (w / v) polysorbate 20, pH 6.0. wherein ARGX-113 is an aqueous formulation comprising an isolated FcRn antigen comprising a variant Fc region. and wherein the variant Fc region comprises two Fc domains that form a homodimer. wherein the amino acid sequence of each of the Fc domains consists of SEQ ID NO:1.

[0153] In one embodiment, the aqueous formulation contains about 100-300 mg / mL of an isolated FcRn antagonist. The solution was diluted with 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and and aqueous solutions containing 0% to 0.04% (w / v) polysorbate 20 or polysorbate 80, pH 6.0. a formulation, wherein the isolated FcRn antagonist comprises a variant Fc region; wherein the variant Fc region is composed of two Fc domains that form a homodimer, and each amino acid sequence of the Fc domain is selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3. become.

[0154] In one embodiment, the aqueous formulation contains 150 mg / mL of the isolated FcRn antagonist in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0 0.04% (w / v) polysorbate 20, pH 6.0, wherein the isolated The FcRn antagonist comprises a variant Fc region, wherein the variant Fc region is and comprising two Fc domains forming a monodimer, wherein the amino acids of each of the Fc domains are The sequence consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0155] In one embodiment, the aqueous formulation contains 165 mg / mL of the isolated FcRn antagonist in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0 0.04% (w / v) polysorbate 20, pH 6.0, wherein the isolated Fc The Rn antagonist comprises a variant Fc region, wherein the variant Fc region is a homozygous Fc region. It consists of two Fc domains that form a dimer, wherein the amino acid sequence of each of the Fc domains is The string consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0156] In one embodiment, the aqueous formulation contains 175 mg / mL of the isolated FcRn antagonist in 20 mL of mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0 0.04% (w / v) polysorbate 20, pH 6.0, wherein the isolated Fc The Rn antagonist comprises a variant Fc region, wherein the variant Fc region is a homozygous Fc region. It consists of two Fc domains that form a dimer, wherein the amino acid sequence of each of the Fc domains is The string consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0157] In one embodiment, the aqueous formulation contains 180 mg / mL of the isolated antagonist in 20 mM Hb. Stidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.04 % (w / v) polysorbate 20, pH 6.0, wherein the isolated FcRn The antagonist comprises a variant Fc region, wherein the variant Fc region is a homodimer. The Fc domains each have an amino acid sequence consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0158] In one embodiment, the aqueous formulation contains 200 mg / mL of the isolated FcRn antagonist. mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0 0.04% (w / v) polysorbate 20, pH 6.0, wherein the isolated Fc The Rn antagonist comprises a variant Fc region, wherein the variant Fc region is a homozygous Fc region. It consists of two Fc domains that form a dimer, wherein the amino acid sequence of each of the Fc domains is The string consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0159] In one embodiment, the aqueous formulation contains 300 mg / mL of the isolated FcRn antagonist in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0 0.04% (w / v) polysorbate 20, pH 6.0, wherein the isolated Fc The Rn antagonist comprises a variant Fc region, wherein the variant Fc region is a homozygous Fc region. It consists of two Fc domains that form a dimer, wherein the amino acid sequence of each of the Fc domains is The string consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0160] In one embodiment, the aqueous formulation comprises about 100-300 mg / mL of ARGX-113 in 20 mM histidine / Histidine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0% to 0.04% (w / v) Polysorbate 20 or Polysorbate 80, an aqueous formulation at pH 6.0, wherein AR GX-113 is an isolated FcRn antagonist consisting of a variant Fc region, wherein the The variant Fc region consists of two Fc domains that form a homodimer, where the Fc domains The main amino acid sequence of each consists of SEQ ID NO:1.

[0161] In one embodiment, the aqueous formulation contains 150 mg / mL ARGX-113 in 20 mM histidine / histidine. gin HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.03% (w / v) Polysol. and a 5'-aminobutyric acid (AgX-113) derivative comprising a variant Fc region of the 5'-aminobutyric acid (AgX-113) and a variant Fc region of the ... and an isolated FcRn antagonist comprising: wherein the amino acid sequence of each of the Fc domains is It consists of sequence number 1.

[0162] In one embodiment, the aqueous formulation comprises about 165 mg / mL of ARGX-113 in 20 mM histidine / histidine. thionine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.04% (w / v) polysorbate The formulation is an aqueous formulation containing ARGX-113 in 1000 mM NaCl, pH 6.0, wherein the ARGX-113 is a variant Fc region. and wherein the variant Fc region is a homodimer. The antibody comprises two Fc domains forming a fusion protein, wherein the amino acid sequence of each of the Fc domains is , consisting of SEQ ID NO:1.

[0163] In one embodiment, the aqueous formulation contains about 165 mg / mL of the isolated FcRn antagonist. 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and and 0.04% (w / v) polysorbate 20, pH 6.0, in which isolated F The cRn antagonist comprises a variant Fc region, wherein the variant Fc region is and comprising two Fc domains forming a monodimer, wherein the amino acids of each of the Fc domains are The sequence consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0164] In one embodiment, the aqueous formulation contains 165 mg / mL ARGX-113 in 20 mM histidine / histidine. gin HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.04% (w / v) Polysol. and a 5'-aminobutyric acid (AgX-113) derivative comprising a variant Fc region of the 5'-aminobutyric acid (AgX-113) and a variant Fc region of the ... and an isolated FcRn antagonist comprising: wherein the amino acid sequence of each of the Fc domains is It consists of sequence number 1.

[0165] In one embodiment, the aqueous formulation contains 175 mg / mL ARGX-113 in 20 mM histidine / histidine. gin HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.03% (w / v) Polysol. and a 5'-aminobutyric acid (AgX-113) derivative comprising a variant Fc region of the 5'-aminobutyric acid (AgX-113) and a variant Fc region of the ... and an isolated FcRn antagonist comprising: wherein the amino acid sequence of each of the Fc domains is It consists of sequence number 1.

[0166] In one embodiment, the aqueous formulation contains 200 mg / mL ARGX-113 in 20 mM histidine / histidine. gin HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.03% (w / v) Polysol. and a 5'-aminobutyric acid (AgX-113) derivative comprising a variant Fc region of the 5'-aminobutyric acid (AgX-113) and a variant Fc region of the ... and an isolated FcRn antagonist comprising: wherein the amino acid sequence of each of the Fc domains is It consists of sequence number 1.

[0167] In one embodiment, the aqueous formulation contains about 100 to about 200 mg / mL of an isolated FcRn antagonist. The stock was prepared from 50 mM histidine / histidine HCl, 60 mM sucrose, 150 mM arginine HCl, and 0. An aqueous preparation containing 0.2% to 0.04% (w / v) polysorbate 20 or polysorbate 80, pH 6.0. wherein the isolated FcRn antagonist consists of a variant Fc region, The variant Fc region is composed of two Fc domains that form a homodimer, wherein the F The amino acid sequence of each of the c domains consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0168] In one embodiment, the aqueous formulation comprises about 100 to about 200 mg / mL of ARGX-113 in 50 mM histidine. 60 mM arginine / histidine HCl, 60 mM sucrose, 150 mM arginine HCl, and 0.02% to 0.04% (w / v) polysaccharide / histidine HCl. An aqueous formulation comprising resorbate 20 or polysorbate 80, pH 6.0, wherein ARGX-1 13 is an isolated FcRn antagonist consisting of a variant Fc region, The ant Fc region consists of two Fc domains that form a homodimer, where the Fc domains The amino acid sequence of each of the peptides consists of SEQ ID NO:1.

[0169] In one embodiment, the aqueous formulation contains about 100 to about 200 mg / mL of an isolated FcRn antagonist. The stock solution was diluted with 50 mM histidine / histidine HCl, 60 mM sucrose, 100 mM arginine HCl, 10 mM L -Methionine and 0.02% to 0.04% (w / v) polysorbate 20 or polysorbate 80, pH 6.0 wherein the isolated FcRn antagonist is a variant Fc region. wherein the variant Fc region is composed of two Fc domains that form a homodimer. wherein the amino acid sequence of each of the Fc domains is selected from SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3. It consists of column number 3.

[0170] In one embodiment, the aqueous formulation contains about 150 mg / mL of the isolated FcRn antagonist. 50mM histidine / histidine HCl, 60mM sucrose, 100mM arginine HCl, 10mM L-methionine and 0.02% to 0.04% (w / v) polysorbate 20, pH 6.0. wherein the isolated FcRn antagonist comprises a variant Fc region, The ant Fc region consists of two Fc domains that form a homodimer, where the Fc domains The amino acid sequence of each of the peptides consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0171] In one embodiment, the aqueous formulation contains about 175 mg / mL of the isolated FcRn antagonist. 50mM histidine / histidine HCl, 60mM sucrose, 100mM arginine HCl, 10mM L-methionine and 0.02% to 0.04% (w / v) polysorbate 20, pH 6.0. wherein the isolated FcRn antagonist comprises a variant Fc region, The ant Fc region consists of two Fc domains that form a homodimer, where the Fc domains The amino acid sequence of each of the peptides consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0172] In one embodiment, the aqueous formulation contains about 200 mg / mL of the isolated FcRn antagonist. 50mM histidine / histidine HCl, 60mM sucrose, 100mM arginine HCl, 10mM L-methionine and 0.02% to 0.04% (w / v) polysorbate 20, pH 6.0. wherein the isolated FcRn antagonist comprises a variant Fc region, The ant Fc region consists of two Fc domains that form a homodimer, where the Fc domains The amino acid sequence of each of the peptides consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.

[0173] In one embodiment, the aqueous formulation comprises about 100 to about 200 mg / mL of ARGX-113 in 50 mM histidine. arginine / histidine HCl, 60 mM sucrose, 100 mM arginine HCl, 10 mM L-methionine, and 0.0 An aqueous preparation containing 2% to 0.04% (w / v) polysorbate 20 or polysorbate 80, pH 6.0. wherein ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc region. wherein the variant Fc region consists of two Fc domains that form a homodimer; Here, the amino acid sequence of each of the Fc domains consists of SEQ ID NO:1.

[0174] In one embodiment, the aqueous formulation comprises about 150 mg / mL of ARGX-113 in 50 mM histidine / histidine. arginine HCl, 60 mM sucrose, 100 mM arginine HCl, 10 mM L-methionine, and 0.03% (w / v ) an aqueous formulation comprising polysorbate 20, pH 6.0, wherein ARGX-113 is variant F an isolated FcRn antagonist consisting of a variant Fc region, wherein the variant Fc region comprises: It consists of two Fc domains that form a homodimer, where each amino acid of the Fc domain is The acid sequence consists of SEQ ID NO:1.

[0175] In one embodiment, the aqueous formulation comprises about 175 mg / mL of ARGX-113 in 50 mM histidine / histidine. arginine HCl, 60 mM sucrose, 100 mM arginine HCl, 10 mM L-methionine, and 0.03% (w / v ) an aqueous formulation comprising polysorbate 20, pH 6.0, wherein ARGX-113 is variant F an isolated FcRn antagonist consisting of a variant Fc region, wherein the variant Fc region comprises: It consists of two Fc domains that form a homodimer, where each amino acid of the Fc domain is The acid sequence consists of SEQ ID NO:1.

[0176] In one embodiment, the aqueous formulation comprises about 200 mg / mL of ARGX-113 in 50 mM histidine / histidine. arginine HCl, 60 mM sucrose, 100 mM arginine HCl, 10 mM L-methionine, and 0.03% (w / v ) an aqueous formulation comprising polysorbate 20, pH 6.0, wherein ARGX-113 is variant F an isolated FcRn antagonist consisting of a variant Fc region, wherein the variant Fc region comprises: It consists of two Fc domains that form a homodimer, where each amino acid of the Fc domain is The acid sequence consists of SEQ ID NO:1.

[0177] In one embodiment, the aqueous formulation comprises about 100-300 mg / mL of isolated neonatal Fc receptor (Fc Rn) antagonists were dissolved in 50 mM histidine / histidine HCl, 200 mM arginine HCl, pH 6.5. wherein the isolated FcRn antagonist is a variant Fc region. wherein the variant Fc region comprises two Fc domains that form a homodimer. wherein the amino acid sequence of each of the Fc domains is SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3. It consists of the number 3.

[0178] In one embodiment, the aqueous formulation contains about 200-300 mg / mL of isolated neonatal Fc receptor (FcRn ) Antagonists were dissolved in 50 mM histidine / histidine HCl, 200 mM arginine HCl, pH 6.5. wherein the isolated FcRn antagonist is an aqueous formulation comprising a variant Fc region. wherein the variant Fc region comprises two Fc domains that form a homodimer. wherein the amino acid sequence of each of the Fc domains consists of SEQ ID NO:1.

[0179] In one embodiment, the aqueous formulation contains about 250-300 mg / mL of isolated neonatal Fc receptor (FcRn ) Antagonists were dissolved in 50 mM histidine / histidine HCl, 200 mM arginine HCl, pH 6.5. wherein the isolated FcRn antagonist is an aqueous formulation comprising a variant Fc region. wherein the variant Fc region comprises two Fc domains that form a homodimer. wherein the amino acid sequence of each of the Fc domains consists of SEQ ID NO:1.

[0180] According to each of the foregoing embodiments of this aspect, in certain embodiments, the device comprises from about 1 to about 2 According to each of the foregoing embodiments of this aspect, in one embodiment Thus, the device contains 1 to 2.5 mL of the aqueous formulation.

[0181] In some embodiments, the device includes a syringe containing the aqueous formulation. Optionally, a suitable syringe may be used to administer at least a portion of the aqueous solution contained in the syringe to a subject. Fine gauge needles (small diameter) cause less pain to the patient. However, it requires a drug with low viscosity. The needle gauge is preferably 27 gauge or larger (i.e., The outer diameter of the needle may be 0.413 mm, 0.41 mm, or smaller.

[0182] In some embodiments, the syringe is provided as a pre-filled syringe. The filled syringes are suitable for single use or alternatively for multiple (two or more) uses. Such pre-filled syringes can optionally contain a small amount of the aqueous solution contained in the syringe. In some embodiments, the catheter may be equipped with a needle suitable for administering at least some of the catheter to a subject. In the present invention, the prefilled syringes are provided in a single unit package.

[0183] In some embodiments, the prefilled syringe is substantially free of air. In such embodiments, the aqueous formulation contained in the prefilled syringe is substantially depleted of dissolved oxygen. For example, the aqueous formulation contained in a prefilled syringe may be administered using the methods described herein. It is then placed in a syringe and filled with nitrogen to remove the air. In one such embodiment, the prefilled syringe can be sealed at ambient temperature. It may be provided in a gas impermeable package.

[0184] In certain embodiments, the present invention provides a method for the preparation of a medicament for use in a pharmaceutical composition comprising, for example, 360 mg / 2 mL (= 180 mg / mL) or 330 mg / 2 mL (= 1 65 mg / mL) of an isolated neonatal Fc receptor (FcRn) antagonist, such as ARGX-113; A pre-filled syringe containing 2 mL or 2.1 mL of the aqueous formulation described herein. The present invention relates to, for example, an isolated neonatal Fc receptor (FcRn) antigen of 360 mg / 2.2 mL (=165 mg / mL). The solution is filled with 2.2 mL of an aqueous formulation described herein containing an antagonist, e.g., ARGX-113. Such vials are suitable for use in kits containing an aqueous solution. At least a portion of the fluid can be coupled with a needle suitable for administering to a subject.

[0185] The present invention further provides an additional formulation comprising more than 2 mL, 2.1 mL, 2.2 mL, or about 2.5 mL of an aqueous formulation according to the present invention. Such devices include, by way of example and not limitation, the present invention. Approximately 1.8mL, 2mL, 2.1mL, 2.2mL, 2.4mL, 2.6mL, 2.8mL, 3mL, 5mL, approximately 10mL, approximately 20mL This includes 2 mL, 2.1 mL, about 50 mL, and about 100 mL of the aqueous formulation according to the present invention. The formulation is administered subcutaneously by using a prefilled syringe containing 1.0 L, 2.2 mL, or 5 mL of the aqueous formulation. It has the advantage that it can be administered by the patient in one shot as an injection. Such a "push" subcutaneous administration takes approximately 12-20 seconds or up to 1 minute. For comparison: The administration of the injection to the subject by a nurse or caregiver may take anywhere from a few minutes to a few hours; An IV infusion of the drug takes approximately 60 minutes. Prefilled syringes are available for self-administration, subcutaneous injection, It can be used as a maintenance dose, providing benefit to the patient.

[0186] The present invention is further illustrated by the following examples, which should not be construed as further limiting. The sequence listing, figures, and all cited throughout this application are incorporated herein by reference in their entirety. The contents of all references, patents, and published patent applications are expressly incorporated herein by reference. can be. EXAMPLES

[0187] (Example) Example 1. Rheological characterization This example describes experiments performed to develop and characterize high concentration formulations of ARGX-113. In particular, the purpose of this series of experiments was to confirm the concentration-viscosity relationship. The experiment was carried out at two temperatures, 5°C and 25°C. Five concentrations of ARGX-113 at the optimal shear rate were Screening and characterization of the rheological profile of ARGX-113 in platform buffer I confirmed the rule.

[0188] Starting with 4-5 mg / mL ARGX-113 in Tris / glycine buffer, pH 7.2, Replace the ferrofluid with sodium phosphate + salt, pH 6.7, to the maximum possible concentration (target 250 mg / mL), and and separately in histidine HCl (HisHCl) + salt at pH 6.0. The viscosity and pH were monitored. The viscoelastic behavior was measured from 0 to 4000 s -1 The shear rate is increased by The highest concentration was studied in HisHCl + salt at pH 6.0. Serial dilutions were performed (6 concentrations) and the UV-absorbance was Degree measurement (A 280 ) was confirmed. Viscosity vs. concentration measurements were performed for 2000s -1 The shear rate was 100 rpm.

[0189] ARGX-113 in sodium phosphate, pH 6.7+ salt precipitated in solution at ∼130 mg / mL.

[0190] The process of increasing the concentration of ARGX-113 in sodium phosphate + salt at pH 6.7 was very slow. ARGX-113 at a higher concentration (~130mg / mL) in sodium phosphate + salt at storage conditions (5°C vs. 25°C) Thus, a reversible solid / liquid phase transition was observed. ARGX-113 at 5°C (~130 mg / mL) showed visible particles that were significantly larger in number and size.

[0191] In contrast, ARGX-113 in HisHCl+salt at pH 6.0 remained in solution up to at least 260 mg / mL. It remained.

[0192] Treatment with increasing concentrations of ARGX-113 in HisHCl+salts at pH 6.0 was rapid.

[0193] In contrast to sodium phosphate buffer + salt, neither precipitation nor phase separation was evident during the concentration increase. was not seen.

[0194] A concentration-dependent sol-gel transition was observed after storage at 5°C. The HisHCl formulation had a concentration of ∼260 mg / mL. At the 260>200>>120mg / mL concentrations, the gelatinous appearance was very pronounced, which could be seen by warming to room temperature or by pipetting. When pipetted it quickly converted to a liquid.

[0195] ARGX-113 is a novel agonist that inhibits acetylcholinesterase activity in HisHCl+ salt at very high concentrations and low temperatures (>180mg / mL, 5°C) at pH 6.0. It appears to have some thixotropic behavior.

[0196] Shear thinning / thickening of ARGX-113 at maximum concentration (267 mg / mL in HisHCl+salt at pH 6.0, 25 °C) , 0~4000s -1 A ramp of shear rates was tested. Representative results are shown in Figure 1. As shown in the figure, ARGX-113 is -1 Shear viscosity is significant in the range of showed no melting or fluidization. -1 was chosen as the shear rate for further testing.

[0197] Representative results of a concentration vs. viscosity study of ARGX-113 in HisHCl+ salt at pH 6.0 are shown in Figure 2. As shown in the figure, the viscosities at 5°C and 25°C and approximately 180 mg / mL are 6 and 18, respectively. The viscosity at 5°C and 25°C at approximately 200 mg / mL is 9 and 33 mPa s, respectively. I did.

[0198] Additional studies will be conducted to evaluate various excipients and pH values ​​of increasing concentrations of ARGX-113 solutions. Starting with 4–5 mg / mL of ARGX-113 in Tris / glycine buffer, pH 7.2, ARGX-113 Exchange the buffer with sodium phosphate + salt, pH 6.7, to a target concentration of 175 mg / mL. Separately, the concentration was increased in HisHCl + salt at pH 6.0. The concentration and pH were monitored during the treatment. Stock solutions of different excipients were prepared to obtain various target formulation compositions. The formulation conditions of 11(11) using excipients and pH were examined for viscosity reduction evaluation at a high concentration of 175 mg / mL. The various formulations that were investigated are shown in Table 1. Table 1. [Table 1] ArgHCl: Arginine HCl SodPhos: Sodium phosphate

[0199] Phosphate formulations could not be formulated above 100 mg / mL due to significant precipitation. All formulations were stored at 5°C for approximately 48 hours to observe phase transitions, if any. All 11 formulations Viscosity measurement at 5℃ for 2000s -1 and viscosity measurements of selected formulations were performed at 25°C. .

[0200] All formulations except the stock remained liquid and clear even after 2 days when stored at 5°C. F11 (SodPhos+NaCl) became turbid and precipitated upon increasing the concentration to ~129 mg / mL. The phosphate formulation formed a clear solution after formulation and storage at 5°C. F9 (pH 6.5 ) The formulation was slightly opaque, which further became slightly clearer at 5°C.

[0201] Viscosity was low (<25 mPa·s) at 5°C for all formulations F1 to F11. F2 and F5 had low viscosity. Sucrose was effectively reduced at 175 mg in 20 mM His / HisHCl, pH 6.0 (F6 and F8). / mL increased the viscosity.

[0202] Example 2. Further rheological characterization This example was conducted to develop and characterize further high concentration formulation candidates of ARGX-113. In particular, the purpose of this series of experiments was to characterize specific characteristics and short-term stability studies. Identify potential high-concentration liquid formulations of ARGX-113 for preclinical toxicology testing and early clinical trials based on It was to do so.

[0203] The compositions of the four aqueous formulations of ARGX-113 examined in this example are shown in Table 2. Table 2. [Table 2]

[0204] Starting with ~136 mg / mL ARGX-113 in 20 mM L-histidine / L-histidine HCl HO, The -113 buffer was exchanged to achieve the target buffer concentration and pH, and then the The concentration was increased beyond the target concentration. Since the material solidifies when the concentration is increased, F14 was increased to 100 mg / mL. This resulted in a formulation concentration of 100 ml after blending with the stock solution. Limited to g / mL.

[0205] Initial characterization of these formulations included measurements of pH, osmolality via freezing point depression, and actual Determination of protein concentration was included. Representative results are shown in Table 3. Table 3. [Table 3]

[0206] Aliquots of each formulation were incubated in an orbital shaker at a target speed of 115 rpm at room temperature and They were subjected to shaking stress in a horizontal position under cold conditions for about 7 days.

[0207] Aliquots of each formulation were frozen / thawed five times in a vertical position from -65°C to room temperature. It was offered to Ikuru.

[0208] As shown in Figure 3, ARGX-113 concentrations were stable under all test conditions. F1=F12; F2=F13; F3=F14; F4=F15. The bars shown in Figure 3 from left to right are: The following are included: F1 initial; F1 shaken at 5°C (shake sh 5); F1 shaken at 25°C (shake sh 2 5); F1 5 cycles of freeze / thaw stress (freeze / thaw F / T); F1 4 weeks at 5°C (4W5); F1 25 ℃ for 4 weeks (4W25); F1 40℃ for 4 weeks (4W40); F1 5℃ for 8 weeks (8w5); F1 25℃ for 8 weeks (8w2 5); F1 40℃ for 8 weeks (8w40); F2 early stage; F2 5℃ shaking (shaking sh 5); F2 25℃ shaking ( shaking sh 25); F2 5 cycles of freeze / thaw stress (freeze / thaw F / T); F2 4 weeks at 5°C (4W5 ); F2 4 weeks at 25℃ (4W25); F2 4 weeks at 40℃ (4W40); F2 8 weeks at 5℃ (8w5); F2 25℃ 8 weeks (8w25); F2 8 weeks at 40°C (8w40); F3 early stage; F3 shaking at 5°C (shaking sh 5); F3 25 F3 at 5°C (shaking sh 25); F3 at 5°C (freeze / thaw F / T); F3 at 5°C (shaking sh 25); 4 weeks (4W5); F3 4 weeks at 25°C (4W25); F3 4 weeks at 40°C (4W40); F3 8 weeks at 5°C (8w5); F 3 8 weeks at 25°C (8w25); F3 8 weeks at 40°C (8w40); F4 early stage; F4 shaking at 5°C (shaking sh 5) F4 Shaking at 25°C (Shaking sh 25); F4 5 cycles of freeze / thaw stress (Freeze / thaw F / T); F 4 4 weeks at 5℃ (4W5); F4 4 weeks at 25℃ (4W25); F4 4 weeks at 40℃ (4W40); F4 8 weeks at 5℃ (8w5); F4 8 weeks at 25℃ (8w25); F4 8 weeks at 40℃ (8w40).

[0209] As shown in Figures 4A-4B, size exclusion chromatography was performed for all tested liquid formulations. No significant differences in aggregation and fragmentation were observed by SEC. As shown in Figure 5C, integrated chip-based capillary electrophoresis (iCE) No significant chemical decomposition was observed. As shown in Figures 6A-6D, the overall visible grain size The number of small and subvisible particles was low upon shaking and freeze / thaw stress. There was no significant difference between polysorbate 20 and polysorbate 80 preparations, and Both polysorbate 20 and polysorbate 80 at 0.04% w / v were resistant to agitation and freeze / thaw stress. suggested that ARGX-113 was equally protective. In Figures 4A-4B, 5A-5C, and 6A-6D: F1=F12; F2=F13; F3=F14; F4=F15. From left to right in Figures 4A-4B, 5A-5C, and 6A-6D. The bars shown are: F1 initial; F1 shaken at 5°C (shake sh 5); F1 25 F1 at 5°C with shaking (shaking sh 25); F1 at 5°C with freeze / thaw stress for 5 cycles (freeze / thaw F / T); F1 at 5°C with shaking (shaking sh 25); 4 weeks (4W5); F1 4 weeks at 25℃ (4W25); F1 4 weeks at 40℃ (4W40); F1 8 weeks at 5℃ (8w5); F 1 8 weeks at 25℃ (8w25); F1 8 weeks at 40℃ (8w40); F2 early stage; F2 shaking at 5℃ (shaking sh 5) F2 Shaking at 25°C (Shaking sh 25); F2 5 cycles of freeze / thaw stress (Freeze / thaw F / T); F 2 4 weeks at 5℃ (4W5); F2 4 weeks at 25℃ (4W25); F2 4 weeks at 40℃ (4W40); F2 8 weeks at 5℃ (8w5); F2 8 weeks at 25℃ (8w25); F2 8 weeks at 40℃ (8w40); F3 early stage; F3 shaking at 5℃ (shaking F3 shaking at 25°C (shaking sh 25); F3 5 cycles of freeze / thaw stress (freeze / thaw F / T); F3 4 weeks at 5℃ (4W5); F3 4 weeks at 25℃ (4W25); F3 4 weeks at 40℃ (4W40); F3 5 ℃ for 8 weeks (8w5); F3 25℃ for 8 weeks (8w25); F3 40℃ for 8 weeks (8w40); F4 early; F4 5℃ F4 Shaking at 25°C (shaking sh 5); F4 Shaking at 25°C (shaking sh 25); F4 5 cycles of freeze / thaw stress F4 (frozen / thawed F / T); F4 4 weeks at 5°C (4W5); F4 4 weeks at 25°C (4W25); F4 4 weeks at 40°C (4W 40); F4 8 weeks at 5℃ (8w5); F4 8 weeks at 25℃ (8w25); F4 8 weeks at 40℃ (8w40).

[0210] Short-term stability data for up to 2 months (8 weeks) indicates that ARGX-113 is 0.8-1.7% after 8 weeks at 40°C by SEC. These results suggest that ARGX-113 has a moderate tendency to aggregate, accompanied by an increase in the aggregation area. The results were dependent on the concentration, pH, and composition of the specific storage conditions. At 40°C, F14 formulation showed greater aggregation compared to F12 containing arginine. , showed greater aggregation compared to F15 at 100 mg / mL concentration at pH 6.0 at 40°C. suggested that ARGX-113 has better stability at pH 6.0 compared to pH 6.5. Fragmentation was below the limit of quantification (LOQ) by SEC.

[0211] In iCE, ARGX-113 showed highly basic and highly acidic species with an early low main peak. However, this decreased rapidly in stability depending on pH and composition, especially at elevated temperatures. The iCE profile showed that the main peak of ARGX-113 in the liquid formulation was stable for 8 weeks at 40°C. The results suggested that the 100mg / mL and 150mg / mL concentrations of 1,2-dichlorophenylalanine (1,2-dichlorophenylalanine) were primarily converted to the acidic variant (~24%). ARGX-113 is stable over a two-month period and the rate and extent of chemical degradation in the liquid formulation is unknown. For all formulations and stability time points tested, the basic peak No significant changes were observed.

[0212] In addition, CE-SDS (Caliper, PerkinElmer) showed no significant changes over the 8-week stability study. Not shown (results not shown).

[0213] All formulations showed comparable initial results, except for formulation F12, which showed many particles (particle cloud) after 8 weeks at 40°C. The solution was free of visible particles after 8 weeks at 5, 25, and 40°C. The color of the solution remained consistent after storage and formulation. Depending on the condition, the color ranged from slightly brown to brown. All preparations were initially as follows: After 8 weeks at 5 and 40°C, a stable target pH (±0.2) was observed.

[0214] As shown in FIG. 7, the overall turbidity of the 150 mg / mL formulation was 15 FNU (formazin nephelometry). In addition, the sodium chloride formulation had a higher creatinine concentration than the arginine-containing formulation. F13 showed a significant increase in turbidity after 4 and 8 weeks at 40°C. In Figure 7: F1=F12; F2=F13; F3=F14; F4=F15. The bars shown in Figure 7 from left to right are: The following are the results: F1 initial; F1 shaken at 5°C (shake sh 5); F1 shaken at 25°C (shake sh sh 25); F1 5 cycles of freeze / thaw stress (freeze / thaw F / T); F1 4 weeks at 5°C (4W5); F 1 4 weeks at 25℃ (4W25); F1 4 weeks at 40℃ (4W40); F1 8 weeks at 5℃ (8w5); F1 8 weeks at 25℃ (8w25); F2 early stage; F2 shaking at 5℃ (shaking sh 5); F2 shaking at 25℃ (shaking sh 25); F2 5 cycles of freeze / thaw stress (freeze / thaw F / T); F2 4 weeks at 5°C (4W5); F2 4 weeks at 25°C (4W25); F2 4 weeks at 40℃ (4W40); F2 8 weeks at 5℃ (8w5); F2 8 weeks at 25℃ (8w25); F3 Initial; F3 shaking at 5°C (shaking sh 5); F3 shaking at 25°C (shaking sh 25); F3 5 cycles of freezing Freeze / thaw stress (freeze / thaw F / T); F3 4 weeks at 5°C (4W5); F3 4 weeks at 25°C (4W25); F3 4 weeks at 4 4 weeks at 0℃ (4W40); F3 8 weeks at 5℃ (8w5); F3 8 weeks at 25℃ (8w25); F4 early stage; F4 5℃ F4 Shaking at 25°C (shaking sh 5); F4 Shaking at 25°C (shaking sh 25); F4 5 cycles of freeze / thaw stress F4 (frozen / thawed F / T); F4 4 weeks at 5°C (4W5); F4 4 weeks at 25°C (4W25); F4 4 weeks at 40°C (4W 40); F4 8 weeks at 5℃ (8w5); F4 8 weeks at 25℃ (8w25).

[0215] From the results obtained in this example, (i) the formulation containing NaCl was more effective than the formulation containing arginine. The increase in turbidity was higher for the 150 mg / mL NaCl formulation at 40° C.; ) Overall number of invisible particles, except for F12, which shows an increase in visible and invisible particles at 40°C. (iii) the initial level of aggregation is high, but the aggregation The rate of increase is moderate after 8 weeks at 40°C; (iv) the arginine formulation shows a significant increase compared to the NaCl formulation. (v) ARGX-113 at concentrations of 150 and 100 mg / mL shows lower aggregation with respect to 8-week stability; It shows good physical stability at 100 mg / mL, but the physical stability at 150 mg / mL at pH 6.0 is poor. (vi) the initial level of the charge variant is high and the main peak is It was concluded that in all liquid formulations, the predominant variant was the acidic variant. showed slightly higher formation of acidic variants compared to the pH 6.0 formulation; and (vii) Polysorbate 20 and polysorbate 80 at 0.04% w / v concentration were used to treat agitation and freeze-thaw stress. were equally effective in protecting ARGX-113 from

[0216] Example 3. pH and Surfactant Optimization This example was conducted to develop and characterize further high concentration formulation candidates of ARGX-113. In particular, the purpose of this series of experiments was to characterize specific characteristics and short-term stability of A candidate high-concentration liquid formulation of ARGX-113 for preclinical toxicology testing and early clinical trials based on efficacy studies The aim of the study was to identify:

[0217] The compositions of the seven aqueous formulations of ARGX-113 examined in this example are shown in Table 4. Table 4. [Table 4]

[0218] Experiments were performed on each of the formulations to determine stability for 2 weeks at 5°C, 25°C, and 40°C, and for 1 week at 5°C. Tested under shaking stress, shaking stress at 25℃ for 1 week, and freeze / thaw stress for 5 cycles did.

[0219] Buffer exchange and concentration increase were performed at the selected pH in 20 mM HisHCl, 100 mM NaCl. Immediately after, appropriate amounts of sucrose and surfactant stocks were added to achieve the target concentrations. The formulations were stored overnight at 5°C and observed for phase transition. Phase separation was observed in F17 (200 mg / mL), After equilibration at room temperature, it returned to the liquid phase. The various formulations were then filtered and separated into separate vials. I put it in the folder.

[0220] After storage at 5°C for 1 week, and also after stirring at 5°C for 5 days, a reversible solid-liquid phase transition was observed. A clear transition was observed in samples F17, F18, and F19. Upon warming / equilibration to room temperature, the formulations The other formulations (F12, F16, F20, and F21) were visibly visibly irritated under these same conditions. It showed no visible solidification / phase transition.

[0221] After storage at 5°C for 2 weeks, a reversible solid-liquid phase transition was observed in samples F17, F18, and F19. Upon warming / equilibration to room temperature, the formulation reverted to a clear liquid solution. 16, F20, and F21) did not show any visible solidification / phase transition under these same conditions.

[0222] After 2 weeks of storage at 25°C, all samples remained in the liquid phase, but many particles were visible in F21. .

[0223] After 2 weeks of storage at 40°C, an increase in opalescence was observed in F13, F18, F19, and F20. Given the results of Example 2, the increase in opalescence observed in F13 was unexpected.

[0224] FIG. 8 shows that there was no significant change in ARGX-113 protein concentration during these stability studies. This shows that.

[0225] FIG. 9 shows the osmolality of the formulations after two weeks of storage at 5° C.

[0226] Figure 10 shows that after 2 weeks of storage at 40°C, F13 and F18 showed a significant increase in turbidity. After 2 weeks of storage at 40°C, the turbidity doubled in F19 and F20.

[0227] 11A-11B show the aggregation results.

[0228] 12A-12C show the results of iCE.

[0229] 13A-13D show the results for subvisible particles.

[0230] In Figures 8, 9, 10, 11A-11B, 12A-12C, 13A-13D: F1=F13; F2=F16; F3=F17; F4=F 18; F5=F19; F6=F20; F7=F21. From left to right in Figures 8, 10, 11A-11B, 12A-12C, and 13A-13D The bars shown in order from right to left are as follows (Note: Some bars in Figures 13A-13D are very (The order of the bars is the same even if the bars are barely visible in the graph): F 1 Initial; F1 shaken at 5°C (shake sh 5); F1 shaken at 25°C (shake sh 25); F1 frozen for 5 cycles Freeze / thaw stress (freeze / thaw F / T); F1 2 weeks at 5°C (2W5); F1 2 weeks at 25°C (2W25); F1 4 2 weeks at 0°C (2W40); F2 initial stage; F2 shaking at 5°C (shaking shaking sh 5); F2 shaking at 25°C (shaking sh 25); F2 5 cycles of freeze / thaw stress (freeze / thaw F / T); F2 2 weeks at 5°C (2W5); F2 2 2 weeks at 5℃ (2W25); F2 2 weeks at 40℃ (2W40); F3 initial stage; F3 shaking at 5℃ (shaking sh 5); F3 Shaking at 25°C (Shaking sh 25); F3 5 cycles of freeze / thaw stress (Freeze / Thaw F / T); F3 5°C (2W5); F3 2 weeks at 25℃ (2W25); F3 2 weeks at 40℃ (2W40); F4 early stage; F4 5℃ F4 Shaking (shaking sh 5); F4 Shaking at 25°C (shaking sh 25); F4 5 cycles of freeze / thaw stress (freezing Freeze / thaw F / T); F4 2 weeks at 5℃ (2W5); F4 2 weeks at 25℃ (2W25); F4 2 weeks at 40℃ (2W40); F5 initial; F5 shaking at 5°C (shaking sh 5); F5 shaking at 25°C (shaking sh 25); F5 5 cycles Freeze / thaw stress (Freeze / thaw F / T); F5 2 weeks at 5°C (2W5); F5 2 weeks at 25°C (2W25); F5 2 weeks at 40°C (2W40); F6 initial stage; F6 shaking at 5°C (shaking sh 5); F6 shaking at 25°C (shaking sh 25); F6 5 cycles of freeze / thaw stress (freeze / thaw F / T); F6 2 weeks at 5°C (2W5); F6 25 ℃ for 2 weeks (2W25); F6 40℃ for 2 weeks (2W40); F7 initial stage; F7 shaking at 5℃ (shaking sh 5); F7 2 Shaking at 5°C (shaking sh 25); F7 5 cycles of freeze / thaw stress (freeze / thaw F / T); F7 5°C F7 at 25℃ for 2 weeks (2W5); F7 at 25℃ for 2 weeks (2W25); F7 at 40℃ for 2 weeks (2W40).

[0231] From the results obtained in this example, (i) 0.02% polysorbate 20 and 0.04% polysorbate 2 0, shake and freeze ARGX-113 regardless of protein concentration (e.g., 150 or 175 mg / mL). (ii) are equally effective at protecting against thawing stress; Polysorbate 80 protects ARGX-113 from shaking and freeze / thaw stress (iii) Polysorbate 20 and Poloxamer 188 were equally effective in preventing ARGX-113 after shaking and (iv) aggregation is concentration-dependent and equally effective at protecting against freeze / thaw stress; F16 and F17 (175 mg / mL and 200 mg / mL, respectively) were stable after 2 weeks of storage at 40°C. , has greater aggregation compared to F13 (150 mg / mL); (v) ARGX-113 concentrations >175 mg / mL , which exhibits a reversible temperature-dependent solid-liquid phase transition; and (vi) the lower pH (5.0 and 5.3) The pH of 5.0 indicates a higher risk of a temperature-dependent solid-liquid phase transition and the pH of 5.0 indicates a higher risk of forming basic species. It was concluded that this represents a possible risk of chemical decomposition and potential for fragmentation.

[0232] Example 4. Further characterization of excipients This example was conducted to develop and characterize further high concentration formulation candidates of ARGX-113. In particular, the purpose of this series of experiments was to identify specific characteristics and short High-concentration liquid formulation of ARGX-113 for preclinical toxicology testing and early clinical trials based on phase stability studies The objective of this study was to identify potential therapeutic candidates.

[0233] The compositions of the seven aqueous formulations of ARGX-113 examined in this example are shown in Table 5. Table 5. [Table 5]

[0234] F22 and F23 were prepared as 2.0 mL aliquots in vials.

[0235] F24 and F25 were prepared as 2.0 mL aliquots in Nuovo Ompi glass syringes.

[0236] F26 and F27 were prepared as 2.0 mL aliquots in BD SCF Neopak glass syringes.

[0237] For F22, F24, and F26, 20 mM HisHCl with or without 10 mM L-methionine , buffer exchange and concentration increase at selected pH in 100 mM ArgCl, followed by bulk Filter and add appropriate amounts of sucrose and surfactant stock to the blended samples. The target concentration was achieved. The blended and unblended formulations were stored overnight at 5°C and observed for phase transition. No phase separation was observed in either the blended or unblended formulations. The blended formulation was then filtered, filled and finished.

[0238] For F23, F25, and F27, 20 mM HisHCl with or without 10 mM L-methionine Buffer exchange and concentration increase were performed at the selected pH in 100 mM NaCl, followed by bulk filtration. For the blended samples, appropriate amounts of sucrose and surfactant stock were added to obtain the target The blended and unblended formulations were stored overnight at 5°C and observed for phase transition. Phase separation was not observed in either the blended or unblended formulations, but was observed in the unblended bulk formulation. The blended formulation was then filtered, filled and finished.

[0239] All formulations are then subjected to specific storage conditions for a specified period of time and are then inspected for appearance, color, clarity, pH, non-visible particles, purity by SE-HPLC, purity by iCE, purity by CE-SDS, viscosity, and breakage The shear and sliding force measurements were analyzed.

[0240] Under the conditions of storage at 5°C for 3 weeks, storage at 25°C for 3 weeks, and storage at 40°C for 3 weeks, all of the formulation remained in the liquid phase.

[0241] Under the conditions of storage at 5°C for 6 weeks, storage at 25°C for 6 weeks, and storage at 40°C for 6 weeks, all The formulations remained in the liquid phase, while some of the solubility in F22 and F26 remained in the liquid phase after storage at 40°C for 6 weeks. The formation of a haze was observed.

[0242] Under the conditions of storage at 5°C for 9 weeks, storage at 25°C for 9 weeks, and storage at 40°C for 9 weeks, all Most of the formulations remained in the liquid phase, but some precipitate was observed in all formulations stored at 40°C for 9 weeks. The lord was observed.

[0243] The protein concentration was measured for each of F22 to F27 by shaking at 5°C, shaking at 25°C, freezing / thawing, and Store at 5°C for 3 weeks, 25°C for 3 weeks, 40°C for 3 weeks, 5°C for 6 weeks, 25°C for 6 weeks storage at 25°C for 9 weeks, at 40°C for 6 weeks, at 5°C for 9 weeks, at 25°C for 9 weeks, and at 40°C for 9 weeks. It was found to be essentially stable under storage conditions of 10 weeks (initial concentration of 10%). within.

[0244] The pH was measured for each of F22 to F27 after shaking at 5°C, shaking at 25°C, freezing / thawing, and storing at 5°C for 3 weeks. storage at 25℃ for 3 weeks, storage at 40℃ for 3 weeks, storage at 5℃ for 6 weeks, storage at 25℃ for 6 weeks, Storage at 0°C for 6 weeks, 5°C for 9 weeks, 25°C for 9 weeks, and 40°C for 9 weeks. It was found to be stable under these conditions.

[0245] Except for F22 and F24 after 6 weeks storage at 40°C and all formulations after 9 weeks storage at 40°C As a result, all formulations at all time periods tested were virtually free of visible particles.

[0246] Osmolality, viscosity, break force, and slip force of various syringe formulations stored at 5°C for 9 weeks is shown in Table 6. Table 6. [Table 6]

[0247] As shown in Figure 14, the increase in turbidity was significant for all formulations stored at 40°C for 9 weeks. The NaCl formulation and the formulation without L-methionine showed slightly higher levels than the ArgCl formulation. The turbidity was low.

[0248] Aggregation was evaluated using SE-HPLC. Figures 15A-15B show the aggregation results. After 1 week of storage, a loss of approximately 1.5-1.9% of the main peak was observed depending on the formulation. 25 (NaCl formulation) had a slightly higher main peak at 40°C compared to F22 and F24 (ArgCl formulations). F26 and F27 (without L-methionine) showed a loss of 1.0% compared to the formulations containing L-methionine. The loss of monomer was mainly due to the high The reasons for this were due to the high molecular weight (HMW) and aggregate formation. F23 and F25 (NaCl formulations) were gCl formulation) at 40°C. (without L-methionine) showed slightly higher HMW at 40°C compared to the formulation with L-methionine. Ta.

[0249] 16A-16C show the iCE results. After 9 weeks of storage at 40° C., all formulations showed comparable The loss of the main peak (29-32%) of β-glucose was observed. A similar increase in the basic and acidic species was observed. .

[0250] 17A-17D show the results for invisible particles. For samples stored at 40° C. for 9 weeks, The data presented may not be reliable due to precipitation in these samples.

[0251] From the results obtained in this example, (i) the ArgCl-free formulation had a 200 mg / kg saturation after storage at 5° C. / mL; (ii) in contrast, NaCl-containing unformulated formulations remain in a liquid state at concentrations of approximately 200 mg / mL (iii) ArgCl-containing formulations do not remain liquid after storage at 40°C. (iv) In contrast, the NaCl-containing formulation showed precipitation after 6 weeks of storage at 40°C. (v) the NaCl-containing formulation exhibits a higher aggregation rate than the ArgCl formulation; and (vi) It was concluded that precipitation of NaCl-containing formulations was observed after storage at 40°C for 9 weeks. .

[0252] Figure 14, 15A~15B, 16A~16C, 17A~17D: F1V=F22; F2V=F23; F1S=F24; F2S=F25; F3S=F2 6; F4S=F27. Bars shown in order from left to right in Figs. 14, 15A-15B, 16A-16C, and 17A-17D. The following are the results (Note: Some bars in Figures 17A-17D are very low, but the bars are The order of the bars is the same even if few are seen in the middle): F1V initial; F1V at 5°C F1V shake (shake sh 5); F1V shake at 25°C (shake sh 25); F1V 5 cycles of freeze / thaw stress ( Frozen / Thawed F / T); F1V 3 weeks at 5°C (3W5); F1V 3 weeks at 25°C (3W25); F1V 3 weeks at 40°C (3W 40); F1V 6 weeks at 5°C (6W5); F1V 6 weeks at 25°C (6W25); F1V 6 weeks at 40°C (6W40); F1V 9 weeks at 5℃ (9W5); F1V 9 weeks at 25℃ (9W25); F1V 9 weeks at 40℃ (9W40); F2V Early; F2 V shaking at 5°C (shaking sh 5); F2V shaking at 25°C (shaking sh 25); F2V 5 cycles of freeze / thaw Stress (freeze / thaw F / T); F2V 3 weeks at 5°C (3W5); F2V 3 weeks at 25°C (3W25); F2V 40°C F2V at 5°C for 3 weeks (3W40); F2V at 5°C for 6 weeks (6W5); F2V at 25°C for 6 weeks (6W25); F2V at 40°C for 6 weeks (6W 40); F2V 9 weeks at 5°C (9W5); F2V 9 weeks at 25°C (9W25); F2V 9 weeks at 40°C (9W40); F1S Initial; F1S shaking at 5°C (shaking sh 5); F1S shaking at 25°C (shaking sh 25); F1S 5 cycles of freezing Freeze / thaw stress (freeze / thaw F / T); F1S 3 weeks at 5°C (3W5); F1S 3 weeks at 25°C (3W25); F1S 3 weeks at 40℃ (3W40); 6 weeks at F1S 5℃ (6W5); 6 weeks at F1S 25℃ (6W25); 6 weeks at F1S 40℃ (6 weeks (6W40); F1S 5℃ for 9 weeks (9W5); F1S 25℃ for 9 weeks (9W25); F1S 40℃ for 9 weeks (9W40) ; F2S initial; F2S shaking at 5℃ (shaking sh 5); F2S shaking at 25℃ (shaking sh 25); F2S 5th Freeze / thaw stress in clones (freeze / thaw F / T); F2S at 5°C for 3 weeks (3W5); F2S at 25°C for 3 weeks (3W 25); F2S 3 weeks at 40°C (3W40); F2S 6 weeks at 5°C (6W5); F2S 6 weeks at 25°C (6W25); F2S 6 weeks at 40°C (6W40); 9 weeks at F2S 5°C (9W5); 9 weeks at F2S 25°C (9W25); 9 weeks at F2S 40°C (9W40); F3S initial; F3S shaking at 5℃ (shaking sh 5); F3S shaking at 25℃ (shaking sh 25); F3S Five cycles of freeze / thaw stress (freeze / thaw F / T); F3S at 5°C for 3 weeks (3W5); F3S at 25°C for 3 weeks weeks (3W25); F3S 40℃ for 3 weeks (3W40); F3S 5℃ for 6 weeks (6W5); F3S 25℃ for 6 weeks (6W25) ; F3S 6 weeks at 40℃ (6W40); F3S 9 weeks at 5℃ (9W5); F3S 9 weeks at 25℃ (9W25); F3S 40 ℃ for 9 weeks (9W40); F4S initial; F4S shaking at 5℃ (shaking sh 5); F4S shaking at 25℃ (shaking sh 2 5); F4S 5 cycles of freeze / thaw stress (freeze / thaw F / T); F4S 3 weeks at 5°C (3W5); F4S 3 weeks at 25℃ (3W25); 3 weeks at 40℃ (3W40); 6 weeks at 5℃ (6W5); 6 weeks at 25℃ (6W25); F4S at 40℃ for 6 weeks (6W40); F4S at 5℃ for 9 weeks (9W5); F4S at 25℃ for 9 weeks (9W25); F4S 9 weeks at 40℃ (9W40).

[0253] Example 5. Further pH optimization studies This example presents a comparison of two preparation methods: Method 1 (pilot) compared to Method 2 (GMP). Method 2 resulted in a more accurate pH compared to preparation 1.

[0254] Some excipients are added to WFI (water for injection), dissolved, and then mixed with different excipients. Formulation buffer was brought to volume. Excipients were added in random order.

[0255] The resulting formulation buffer is used in the UF / DF (ultrafiltration / diafiltration) formulation process of the protein (ARGX-113). Polysorbate 20 had not yet been added at this stage. Table 7. [Table 7]

[0256] Polysorbate was then added via a 10% solution at a dilution of 996:4 (e.g., for a 1000 kg preparation). 4008 ml of excipient buffer was added to the product formulation buffer. Thus, polysorbate 20 (PS20) was added after the diafiltration / ultrafiltration step. .

[0257] This resulted in the following final ARGX-113 formulation: 20 mM L-histidine / L-histidine hydrochloride, 100 mM salt containing 0.04% (w / v) polysorbate 20 165 mg / mL ARGX-113 in sodium chloride, 60 mM sucrose, 10 mM L-methionine, pH 6.0.

[0258] Using the method of this example, protein (ARGX-113) concentrations greater than 165 mg / mL, e.g., 18 It will be appreciated by those skilled in the art that formulations of 0 mg / mL, 200 mg / mL, or even 300 mg / mL can also be made. Thus, it is understood.

[0259] Example 6. Ultra-high concentration formulation In this example, additional experiments were performed to determine whether the ARG concentration was increased to even higher concentrations, for example, 250 to 300 mg / mL. The potential for a viscosity reducing formulation of X-113 was evaluated. Three formulations of strengths were prepared. The three target formulations are shown in Table 8. Table 8. Target formulation [Table 8]

[0260] Materials and Methods The stock solution of ARGX-113 was subjected to buffer exchange and then increased in concentration to obtain the protein. The (ARGX-113) concentration was measured, diluted to approximately 250 mg / mL, and excipients were added. The samples were further divided into 5°C and 25°C storage lots, and then the viscosity, osmolality, and appearance were evaluated. Inspection and filtration test using a 0.22 μm filter were conducted periodically over a 14-day period. Analyses were performed on days 0 (D0; preparation day), 3 (D3), 7 (D7), and 14 (D14). I did.

[0261] (result) The observed pH, osmolality, protein concentration, viscosity, and appearance are shown in Table 9. do. Table 9. [Table 9] nd: Not implemented

[0262] To summarize: Three formulations of ARGX-113 at concentrations ≥ 250 mg / mL were prepared with 200 mM arginine. The viscosity of the three formulations decreased with pH value. The F103 formulation was stable at 5 and 25°C for 2 weeks. It was the formulation with the lowest viscosity (measured at pH = 6.5, 24 mPa·s, and 5°C). · The F101 formulation could not be prepared uniformly on a small scale due to the formation of gel particles. The F102 formulation showed unexpected variability in viscosity, which appeared to decrease over time. It seemed like. The F103 formulation showed a completely reproducible low viscosity within 2 weeks. The appearance of preserved formulations F102 and F103 remains stable for 2 weeks regardless of whether they are stored at 5°C or 25°C. The situation remained the same throughout. A whitish, highly viscous solution was observed in the stock solution containing 100 mM arginine after 7 days. After 14 days, a solid gel was observed (data not shown). Filtration tests were performed: after filtration at 5°C, the viscosity of F102 and F103 remained low (<26mP a·s). The present application provides the following aspects of the invention. (Aspect 1) Approximately 100-300 mg / mL of isolated neonatal Fc receptor (FcRn) antagonist was added to 20-60 mM histidine. Thyidine / histidine HCl, 0-70 mM sucrose, 0-150 mM NaCl, 0-250 mM arginine HCl, 0 %~0.05% (w / v) polysorbate 20 or polysorbate 80, 0~15mM L-methionine, pH 6. 0 to 6.5, wherein the isolated FcRn antagonist is a variant Fc region. wherein the variant Fc region comprises two Fc domains that form a homodimer. wherein the amino acid sequence of each of the Fc domains is SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3. The aqueous preparation comprises No. 3. (Aspect 2) Approximately 100-200 mg / mL of the isolated neonatal Fc receptor (FcRn) antagonist is dissolved in 20 mM histidine. The solution consisted of 10 mM lysine / histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.02% to 0.04% (w / v) polyisobutene. and in polysorbate 20 or polysorbate 80, pH 6.0, wherein each of the Fc domains is 2. The aqueous formulation of claim 1, wherein the amino acid sequence consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3. (Aspect 3) ARGX-113 at 150 mg / mL was dissolved in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl and 0.04% (w / v) polysorbate 20, pH 6.0, wherein ARGX-113 is wherein the amino acid sequence of each of the Fc domains is SEQ ID NO: 2. The aqueous formulation according to claim 1, comprising No. 1. (Aspect 4) ARGX-113 at 175 mg / mL was dissolved in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl and 0.04% (w / v) polysorbate 20, pH 6.0, wherein ARGX-113 is wherein the amino acid sequence of each of the Fc domains is SEQ ID NO: 2. The aqueous formulation according to claim 1, comprising No. 1. (Aspect 5) ARGX-113 at 200 mg / mL was dissolved in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl and 0.04% (w / v) polysorbate 20, pH 6.0, wherein ARGX-113 is wherein the amino acid sequence of each of the Fc domains is SEQ ID NO: 2. The aqueous formulation according to claim 1, comprising No. 1. (Aspect 6) Approximately 100-200 mg / mL of the isolated neonatal Fc receptor (FcRn) antagonist is dissolved in 20 mM histidine. Thysidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.02% ~0.04% (w / v) polysorbate 20 or polysorbate 80, pH 6.0, wherein said F The amino acid sequence of each of the c domains consists of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3. 1. An aqueous preparation as described in claim 1. (Aspect 7) Approximately 165 mg / mL of the isolated neonatal Fc receptor (FcRn) antagonist was dissolved in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.04% (w / v) poly resorbate 20, pH 6.0, wherein the amino acid sequence of each of the Fc domains is 2. The aqueous formulation of embodiment 1, comprising sequence number 1, sequence number 2, or sequence number 3. (Aspect 8) ARGX-113 at 175 mg / mL was dissolved in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl , 10 mM L-methionine, and 0.03% (w / v) polysorbate 20, pH 6.0, where AR GX-113 is the isolated FcRn antagonist, wherein each of the agonists of the Fc domain 2. The formulation of embodiment 1, wherein the amino acid sequence consists of SEQ ID NO:1. (Aspect 9) ARGX-113 at 200 mg / mL was dissolved in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl , 10 mM L-methionine, and 0.03% (w / v) polysorbate 20, pH 6.0, where AR GX-113 is the isolated FcRn antagonist, wherein each of the agonists of the Fc domain 2. The formulation of embodiment 1, wherein the amino acid sequence consists of SEQ ID NO:1. (Aspect 10) Approximately 100-200 mg / mL of the isolated neonatal Fc receptor (FcRn) antagonist is dissolved in 50 mM histidine. Thysidine / histidine HCl, 60 mM sucrose, 150 mM arginine HCl, and 0.02% to 0.04% (w / v ) in polysorbate 20 or polysorbate 80, pH 6.0, wherein the Fc domain 2. The method according to claim 1, wherein each of the amino acid sequences consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3. Sexual preparations. (Aspect 11) Approximately 100-200 mg / mL of the isolated neonatal Fc receptor (FcRn) antagonist is dissolved in 20 mM histidine. Thyidine / histidine HCl, 60 mM sucrose, 100 mM arginine HCl, 10 mM L-methionine, and and 0.02% to 0.04% (w / v) polysorbate 20 or polysorbate 80, pH 6.0, wherein the amino acid sequence of each of the Fc domains is selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3. 2. The aqueous formulation of embodiment 1, comprising: (Aspect 12) ARGX-113 at 175 mg / mL was dissolved in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM arginine, and 100 mM glycerol. in 10 mM L-methionine HCl, 10 mM L-methionine, and 0.03% (w / v) polysorbate 20, pH 6.0; wherein ARGX-113 is said isolated FcRn antagonist, and wherein said Fc domain The amino acid sequence of each of the above consists of SEQ ID NO:1. (Aspect 13) ARGX-113 at 200 mg / mL was dissolved in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM arginine, and 100 mM glycerol. in 10 mM L-methionine HCl, 10 mM L-methionine, and 0.03% (w / v) polysorbate 20, pH 6.0; wherein ARGX-113 is said isolated FcRn antagonist, and wherein said Fc domain 2. The aqueous formulation of embodiment 1, wherein the amino acid sequence of each of the above consists of SEQ ID NO:1. (Aspect 14) Approximately 100 to 300 mg / mL of the isolated neonatal Fc receptor (FcRn) antagonist is dissolved in 50 mM histidine. in 200 mM arginine / histidine HCl, pH 6.5, wherein the Fc domain 2. The method according to claim 1, wherein each of the amino acid sequences consists of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3. Aqueous formulation. (Aspect 15) A packaged product comprising a sterile container containing a therapeutically effective amount of the aqueous formulation according to any one of aspects 1 to 14. Pharmaceutical products. (Aspect 16) A device comprising a therapeutically effective amount of the aqueous formulation according to any one of embodiments 1 to 14. (Aspect 17) 17. The device of embodiment 16, wherein the device comprises a syringe containing the aqueous formulation.

Claims

1. Approximately 100-300 mg / mL of neonatal Fc receptor (FcRn) antagonist, 20-60 mM histidine / histidine 0-70 mM sucrose, 0-150 mM NaCl, 0-250 mM arginine HCl, 0%-0.05% (w / v) An aqueous formulation containing polysorbate 20 or polysorbate 80 and 0-15 mM L-methionine. the aqueous formulation has a pH of 6.0 to 6.5, and the FcRn antagonist is a variant Fc region. wherein the variant Fc region comprises two Fc domains that form a homodimer. wherein the amino acid sequence of each of the Fc domains is the amino acid sequence set forth in SEQ ID NO: 1 or SEQ ID NO:

2. The aqueous preparation comprises an amino acid sequence.

2. About 100-200 mg / mL of the FcRn antagonist, 20 mM histidine / histidine HCl, 60 mM sucrose cellulose, 100 mM NaCl, and 0.02% to 0.04% (w / v) polysorbate 20 or polysorbate 8 10. The aqueous formulation of claim 1, wherein the aqueous formulation has a pH of 6.

0.

3. 150 mg / mL of the FcRn antagonist, 20 mM histidine / histidine HCl, 60 mM sucrose , 100 mM NaCl, and 0.04% (w / v) polysorbate 20, wherein the aqueous formulation has a pH of 6.0 and the amino acid sequence of each of the Fc domains of the FcRn antagonist is SEQ ID NO:

2. The aqueous formulation of claim 1, comprising the amino acid sequence set forth in claim 1.

4. 175 mg / mL of the FcRn antagonist, 20 mM histidine / histidine HCl, 60 mM sucrose , 100 mM NaCl, and 0.04% (w / v) polysorbate 20, wherein the aqueous formulation has a pH of 6.0 and the amino acid sequence of each of the Fc domains of the FcRn antagonist is SEQ ID NO:

2. The aqueous formulation of claim 1, comprising the amino acid sequence set forth in claim 1.

5. 200 mg / mL of the FcRn antagonist, 20 mM histidine / histidine HCl, 60 mM sucrose , 100 mM NaCl, and 0.04% (w / v) polysorbate 20, wherein the aqueous formulation has a pH of 6.0 and the amino acid sequence of each of the Fc domains of the FcRn antagonist is SEQ ID NO:

2. The aqueous formulation of claim 1, comprising the amino acid sequence set forth in claim 1.

6. About 100-200 mg / mL of the FcRn antagonist, 20 mM histidine / histidine HCl, 60 mM sucrose cellulose, 100 mM NaCl, 10 mM L-methionine, and 0.02% to 0.04% (w / v) polysorbate 20 or polysorbate 80, wherein the aqueous formulation has a pH of 6.

0. Aqueous formulation.

7. About 165 mg / mL of the FcRn antagonist, 20 mM histidine / histidine HCl, 60 mM sucrose 100 mM NaCl, 10 mM L-methionine, and 0.04% (w / v) polysorbate 20, 2. The aqueous formulation of claim 1, wherein the aqueous formulation has a pH of 6.

0.

8. 175 mg / mL of the FcRn antagonist, 20 mM histidine / histidine HCl, 60 mM sucrose , 100 mM NaCl, 10 mM L-methionine, and 0.03% (w / v) polysorbate 20, The aqueous formulation has a pH of 6.0 and each of the Fc domains of the FcRn antagonist is 2. The aqueous preparation according to claim 1, wherein the amino acid sequence consists of the amino acid sequence set forth in SEQ ID NO:

1.

9. 200 mg / mL of the FcRn antagonist, 20 mM histidine / histidine HCl, 60 mM sucrose , 100 mM NaCl, 10 mM L-methionine, and 0.03% (w / v) polysorbate 20, The aqueous formulation has a pH of 6.0 and each of the Fc domains of the FcRn antagonist is 2. The aqueous preparation according to claim 1, wherein the amino acid sequence consists of the amino acid sequence set forth in SEQ ID NO:

1.

10. About 100-200 mg / mL of the FcRn antagonist, 50 mM histidine / histidine HCl, 60 mM sucrose cellulose, 150 mM arginine HCl, and 0.02% to 0.04% (w / v) polysorbate 20 or polysorbate 2. The aqueous formulation of claim 1, comprising Rubate 80, wherein the aqueous formulation has a pH of 6.

0.

11. About 100-200 mg / mL of the FcRn antagonist, 20 mM histidine / histidine HCl, 60 mM sucrose cellulose, 100 mM arginine HCl, 10 mM L-methionine, and 0.02% to 0.04% (w / v) Polysol. and wherein the aqueous formulation has a pH of 6.

0. Item 1. The aqueous formulation according to item 1.

12. 175 mg / mL of the FcRn antagonist, 20 mM histidine / histidine HCl, 60 mM sucrose , 100 mM arginine HCl, 10 mM L-methionine, and 0.03% (w / v) polysorbate 20; wherein the aqueous formulation has a pH of 6.0 and 2. The aqueous formulation according to claim 1, wherein each amino acid sequence consists of the amino acid sequence set forth in SEQ ID NO:

1. 。

13. 200 mg / mL of the FcRn antagonist, 20 mM histidine / histidine HCl, 60 mM sucrose , 100 mM arginine HCl, 10 mM L-methionine, and 0.03% (w / v) polysorbate 20; wherein the aqueous formulation has a pH of 6.0 and 2. The aqueous formulation according to claim 1, wherein each amino acid sequence consists of the amino acid sequence set forth in SEQ ID NO:

1. 。

14. About 100-300 mg / mL of the FcRn antagonist, 50 mM histidine / histidine HCl and 200 mM 2. The aqueous formulation of claim 1, comprising arginine HCl, wherein the aqueous formulation has a pH of 6.

5. Agent.

15. 100-200mg / mL FcRn antagonist, 20mM L-histidine / L-histidine HCl, 100mM Na Cl, 60 mM sucrose, 10 mM L-methionine, and 0.04% (w / v) polysorbate 20. The formulation, wherein the aqueous formulation has a pH of 6.0 to 6.5 and the FcRn antagonist is a variant FcRn antagonist. wherein the variant Fc region is composed of two Fc domains that form a homodimer. wherein the amino acid sequence of each of the Fc domains is set forth in SEQ ID NO:1 or SEQ ID NO:

2. The aqueous preparation comprises an amino acid sequence as follows:

16. The amino acid sequence of each of the Fc domains consists of the amino acid sequence set forth in SEQ ID NO:

1.

16. The aqueous formulation of claim 15.

17. 17. The aqueous formulation of claim 15 or 16, comprising 180 mg / mL of said FcRn antagonist.

18. A packaged pharmaceutical product comprising a sterile container containing the aqueous formulation according to any one of claims 1 to 17. 。

19. A device comprising the aqueous formulation according to any one of claims 1 to 17.

20. 20. The device of claim 19, wherein the device comprises a syringe containing the aqueous formulation.

21. The sterile container comprises a single-use vial, a multi-use vial, or a pre-filled syringe; 20. The packaged pharmaceutical product of claim 18.

22. 20. The packaged pharmaceutical product of claim 18, wherein the sterile container comprises a single-use vial.

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