Pharmaceutical formulations of FcRn inhibitors suitable for subcutaneous administration

Abstract

Provided are various water-soluble formulations of the neonatal Fc receptor (FcRn) antagonist ARGX-113, which include formulations useful as pharmaceutical compositions, methods of preparing the formulations, devices comprising the various formulations, and uses thereof. In some embodiments, the formulations are suitable and useful for administering ARGX-113 to human subjects. In some embodiments, the formulations are suitable and useful for administering ARGX-113 subcutaneously to human subjects. These formulations may be used to treat any condition in which inhibition of FcRn-mediated antibody recycling would be beneficial. Such conditions may include one or more of various antibody-mediated autoimmune diseases, including, for example and without limitation, myasthenia gravis (MG) and immune thrombocytopenia (ITP).

Description

Background Technology

[0001] Immunoglobulin gamma (IgG) antibodies play a major role in the pathology of many diseases, such as autoimmune diseases, inflammatory diseases, and pathological diseases characterized by the overexpression of IgG antibodies (e.g., hypergammaglobulinemia) (Junghans, Immunol Res . 16 (1):29 (1997) see also)).

[0002] The half-life of IgG in serum is relatively long compared to the serum half-life of other plasma proteins (Roopenian et al., J Immunol . 170:3528 (2003); Junghans and Anderson, Proc. Natl . Acad . Sci USA 93:5512 (1996)). This long half-life is, in part, due to the binding of the Fc site of IgG to the neonatal Fc receptor (FcRn). Although FcRn was originally identified as the neonatal transport receptor for maternal IgG, it also functions to protect against the degradation of IgG in adults. FcRn binds to pinocytosed IgG and recycles the IgG into the extracellular region, preventing the IgG from being transported to degradable lysosomes. This recycling is facilitated by the pH-dependent binding of IgG to FcRn, where the IgG / FcRn interaction is stronger at acidic endosomal pH than at extracellular physiological pH.

[0003] When serum IgG concentrations reach levels exceeding available FcRn molecules, unbound IgG is not protected from degradation mechanisms and therefore will have a reduced serum half-life. Consequently, inhibition of IgG binding to FcRn reduces the serum half-life of IgG by preventing the recycling of IgG endosomes. Thus, agents that antagonize IgG binding to FcRn may be useful for controlling, treating, or preventing antibody-mediated disorders such as autoimmune diseases and inflammatory diseases. Some of these diseases, at least in part, are currently treated with intravenous infusion of pooled IgG (IVIg) collected from human donors. Since many of these autoimmune diseases are chronic, affected individuals may require repeated IVIg administration and / or other appropriate treatments to manage the disease.

[0004] As another approach, blocking antibodies against FcRn was developed to inhibit the binding of IgGFc to FcRn (see, e.g., WO 2002 / 043658). Additionally, peptides that bind to FcTn and antagonize FcRn function were identified (see, e.g., US 6,212,022 and US 8,101,186). Furthermore, full-length IgG antibodies containing variant Fc receptors with enhanced FcRn binding and reduced pH dependence were also identified, which antagonize the binding of FcRn to Ig (see, e.g., US 8,163,881 and Vaccaro et al. Nat Biotechnol . 23(10): 1283-1288 (2005)).

[0005] Recently, a modified version of the human IgG1 Fc fragment, called another FcRn inhibitor, efgartigimod (also known as ARGX-113), was developed. See WO 2015 / 100299, the full text of which is incorporated here. Clinical trials for ARGX-113 are currently underway in many autoimmune diseases, including myasthenia gravis (MG) and immune thrombocytopenia (ITP).

[0006] There is still a need for improved formulations and methods of administering FcRn inhibitors for use in the treatment of autoimmune diseases.

[0007] Disclosed herein are various formulations of ARGX-113, including formulations useful as pharmaceutical compositions, methods for manufacturing the same, devices comprising the various formulations, and the use thereof. In some embodiments, these formulations are suitable and useful for administering ARGX-113 to human subjects. In some embodiments, these formulations are suitable and useful for administering ARGX-113 subcutaneously to human subjects. These formulations may be used to treat any condition that may benefit from inhibiting FnRn-mediated antibody recycling. Such conditions may include one or more of various antibody-mediated autoimmune diseases, including, for example and without limitation, myasthenia gravis (MG) and immune thrombocytopenia (ITP).

[0008] One aspect of the present invention is a water-soluble formulation comprising about 100-300 mg / mL of isolated neonatal Fc receptor (FcRn) antagonist in 20-60 mM histidine / 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, 0-15 mM L-methionine, and pH 6.0-6.5, wherein the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is SEQ ID NO. 1, It consists of sequence number 2, or sequence number 3.

[0009] In one embodiment, the water-soluble formulation comprises about 100-300 mg / mL of isolated neonatal Fc receptor (FcRn) antagonist in 20-60 mM histidine / histidine HCl, 0-70 mM sucrose, about 100 mM NaCl, 0%-0.05% (w / v) polysorbate 20 or polysorbate 80, 0-15 mM L-methionine, and pH 6.0-6.5, wherein the isolated neonatal FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0010] In one embodiment, the water-soluble formulation comprises about 100-300 mg / mL of isolated neonatal Fc receptor (FcRn) antagonist in 20-60 mM histidine / histidine HCl, 0-70 mM sucrose, 100-250 mM arginine HCl, 0%-0.05% (w / v) polysorbate 20 or polysorbate 80, 0-15 mM L-methionine, and pH 6.0-6.5, wherein the isolated neonatal FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is SEQ ID NO. 1, SEQ ID NO. 2, or It consists of sequence number 3.

[0011] In one embodiment, the water-soluble formulation comprises about 100-300 mg / mL of isolated neonatal Fc receptor (FcRn) antagonist in 20-60 mM histidine HCl, 100-250 mM arginine HCl, 0%-0.05% (w / v) polysorbate 20 or polysorbate 80, 0-15 mM L-methionine, and pH 6.0-6.5, wherein the isolated neonatal FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0012] In each aspect and embodiment mentioned above, in some embodiments, the water-soluble formulation contains 20 or 50 mM histidine.

[0013] In each aspect and embodiment mentioned above, in some embodiments, the water-soluble formulation comprises 0.02%-0.04 (w / v) polysorbate 20 or polysorbate 80.

[0014] In each aspect and embodiment mentioned above, in some embodiments, the water-soluble formulation contains 0 or 10 mM L-methionine.

[0015] In each of the aforementioned aspects and embodiments, in some embodiments, the pH is 6.0 or 6.5.

[0016] In each aspect and embodiment mentioned above, in some embodiments, the amino acid sequence of each Fc domain consists of sequence number 1.

[0017] One aspect of the present invention is a water-soluble formulation comprising about 100-200 mg / mL of isolated neonatal Fc receptor (FcRn) antagonist in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.02%-0.04% (w / v) polysorbate 20 or polysorbate 80, pH 6.0, wherein the isolated FcRn antagonist is composed of a variant Fc site, said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0018] In one embodiment, the water-soluble formulation comprises about 100-200 mg / mL ARGX-113 in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.02%-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 site, the aforementioned variant Fc site consists of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0019] One aspect of the present invention is a water-soluble formulation comprising about 150 mg / mL ARGX-113 in 20 mM histidine / histidineHCl, 60 mM 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 site, the said variant Fc site consists of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0020] One aspect of the present invention is a water-soluble formulation comprising 165 mg / mL ARGX-113 in 20 mM histidine / histidineHCl, 60 mM sucrose, 100 mM NaCl, and about 0.04% (w / v) polysorbate 20, pH 6.0, wherein ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc site, the said variant Fc site consists of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0021] One aspect of the present invention is a water-soluble formulation comprising 175 mg / mL ARGX-113 in 20 mM histidine / histidineHCl, 60 mM sucrose, 100 mM NaCl, and about 0.04% (w / v) polysorbate 20, pH 6.0, wherein ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc site, the said variant Fc site consists of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0022] One aspect of the present invention is a water-soluble formulation comprising 180 mg / mL ARGX-113 in 20 mM histidine / histidineHCl, 60 mM sucrose, 100 mM NaCl, and about 0.04% (w / v) polysorbate 20, pH 6.0, wherein ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc site, the said variant Fc site consists of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0023] One aspect of the present invention is a water-soluble formulation comprising 200 mg / mL ARGX-113 in water, 20 mM histidine / histidineHCl, 60 mM 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 site, the said variant Fc site consists of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0024] One aspect of the present invention is a water-soluble formulation comprising about 100-200 mg / m of isolated neonatal Fc receptor (FcRn) antagonist in 20 mM histidine / histidineHCl, 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 the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0025] In one embodiment, the water-soluble formulation comprises about 100-200 mg / mL of ARGX-113 in 20 mM histidine / histidineHCl, 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 ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc site, the aforementioned variant Fc site consists of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0026] One aspect of the present invention is a water-soluble formulation comprising 165 mg / mL of isolated neonatal Fc receptor (FcRn) antagonist in 20 mM histidine / histidineHCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.04% (w / v) polysorbate 20, pH 6.0, wherein the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0027] In one embodiment, the water-soluble formulation comprises about 165 mg / mL of ARGX-113 in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.02%-0.04% (w / v) polysorbate 20, pH 6.0, wherein ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc site, the aforementioned variant Fc site consists of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0028] One aspect of the present invention is a water-soluble formulation comprising 175 mg / mL ARGX-113 in 20 mM histidine / histidineHCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.03% (w / v) polysorbate 20, pH 6.0, wherein ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc site, the aforementioned variant Fc site consists of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0029] One aspect of the present invention is a water-soluble formulation comprising 180 mg / mL ARGX-113 in 20 mM histidine / histidineHCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.03% (w / v) polysorbate 20, pH 6.0, wherein ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc site, the aforementioned variant Fc site consists of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0030] One aspect of the present invention is a water-soluble formulation comprising 200 mg / mL ARGX-113 in 20 mM histidine / histidineHCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.03% (w / v) polysorbate 20, pH 6.0, wherein ARGX-113 is an isolated FcRn antagonist composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1.

[0031] One aspect of the present invention is a water-soluble formulation comprising about 100-200 mg / mL of isolated neonatal Fc receptor (FcRn) antagonist in 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 is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0032] In one embodiment, the water-soluble formulation comprises about 100-200 mg / mL ARGX-113 in 50 mM histidine / histidine HCl, 60 mM sucrose, 100 mM arginine HCl, and 0.02-0.04% (w / v) polysorbate 20 or polysorbate 80, pH 6.0, wherein ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc site, the aforementioned variant Fc site consists of an Fc domain homomer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0033] One aspect of the present invention is a water-soluble formulation comprising about 100-200 mg / mL of isolated neonatal Fc receptor (FcRn) antagonist in 20 mM histidine / histidine 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 the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0034] In one embodiment, the water-soluble formulation comprises about 100-200 mg / mL ARGX-113 in 20 mM histidine / histidine 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-113 is an isolated FcRn antagonist consisting of a variant Fc site, the aforementioned variant Fc site consists of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0035] One aspect of the present invention is a water-soluble formulation comprising 175 mg / mL ARGX-113 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 ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc site, the aforementioned variant Fc site consists of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0036] One aspect of the present invention is a water-soluble formulation comprising 180 mg / mL ARGX-113 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 ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc site, the aforementioned variant Fc site consists of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0037] One aspect of the present invention is a water-soluble formulation comprising 200 mg / mL ARGX-113 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 ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc site, the aforementioned variant Fc site consists of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0038] One aspect of the present invention is a water-soluble formulation comprising 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM arginine HCl, 10 mM L-methionine, and 0.03% (w / v) polysorbate 20 having a pH of about 6.0, with 250 mg / mL, 300 mg / mL, or more than 300 mg / mL of ARGX-113, wherein ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc site, wherein the aforementioned variant Fc site consists of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0039] One aspect of the present invention is a water-soluble formulation comprising about 100-300 mg / mL of isolated neonatal Fc receptor (FcRn) antagonist in about 200 mM arginine HCl, 50 mM histidine / histidine HCl, having a pH of 6.5, wherein the isolated FcRn antagonist is composed of a variant Fc site, wherein the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0040] In one embodiment, the water-soluble formulation comprises about 200-300 mg / mL of isolated neonatal Fc receptor (FcRn) antagonist in 50 mM histidine / histidine HCl and 200 mM arginine HCl having a pH of 6.5, wherein the isolated FcRn antagonist is composed of a variant Fc site, wherein the aforementioned variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1.

[0041] In one embodiment, the water-soluble formulation comprises about 250-300 mg / mL of isolated neonatal Fc receptor (FcRn) antagonist in 50 mM histidine / histidine HCl and 200 mM arginine HCl having a pH of 6.5, wherein the isolated FcRn antagonist is composed of a variant Fc site, wherein the aforementioned variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1.

[0042] In any embodiment of each prior art aspect of the present invention, the water-soluble formulation is substantially free of dissolved oxygen.

[0043] In any embodiment of each prior art aspect of the present invention, the water-soluble formulation is suitable for in vivo use.

[0044] In any embodiment of each prior art aspect of the present invention, the water-soluble formulation is suitable for use subcutaneously in vivo.

[0045] One aspect of the present invention is a packaged pharmaceutical product comprising a sterile container containing a therapeutically effective amount of a water-soluble formulation of any one of the prior art aspects and embodiments.

[0046] One aspect of the present invention is a device comprising a therapeutically effective amount of a water-soluble formulation of any one of the prior art aspects and embodiments.

[0047] In some embodiments, the device comprises or is configured as a syringe containing a water-soluble formulation.

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

[0049] Fig. 1 is 0-4000 s -1 It shows the shear thinning / thickening behavior of ARGX-113 at the highest concentration studied by shear rate ramping. Figure 2 shows the viscosity (mPa·s) of ARGX-113 versus concentration (mg / mL) in HisHCl + salt (Salt) at the presented concentration and temperature (°C). Figure 3 shows the protein concentration of the ARGX-113 formulation described in Example 2 as measured by UV / Vis. 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; 8w40, 8 weeks at 40 °C. Figures 4A and 4B show the size exclusion chromatography (SEC) results for the main peak and high molecular weight (HMW) species 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; 8w40, 8 weeks at 40 °C. Figures 5A–5C show the chemical degradation measured by iCE for the major 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; 8w40, 8 weeks at 40 °C. FIGS. 6A-6D show subvisible particles of the ARGX-113 formulation described in Example 2, with diameters ≥2 μm, ≥5 μm, ≥10 μm, and ≥25 μm, 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; 8w40, 8 weeks at 40 °C. Figure 7 shows the turbidity of the ARGX-113 formulation described in Example 2. Figure 8 shows the protein concentration as measured by UV / Vis of the ARGX-113 formulation 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, 2 weeks at 40 °C. Figure 9 shows the osmolality as measured by the freezing point depression of the ARGX-13 formulation described in Example 3. Figure 10 shows the turbidity of the ARGX-113 formulation 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, 2 weeks at 40 °C. Figures 11A and 11B show the size exclusion chromatography (SEC) results for the main peak and high molecular weight (HMW) species 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, 2 weeks at 40 °C. FIGS. 12A-12C show the chemical degradation measured by iCE for the major 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, 2 weeks at 40 °C. FIGS. 13A-13D show subvisible particles of the ARGX-113 formulation described in Example 3, with diameters ≥2 μm, ≥5 μm, ≥10 μm, and ≥25 μm, respectively. sh5, shaking at 5 °C; sh25, shaking at 25 °C; 2w5, 2 weeks at 5 °C; 2w25, 2 weeks at 25 °C; 2w40, 2 weeks at 40 °C. Figure 14 shows the turbidity of the ARGX-113 formulation described in Example 4. sh5, shaking at 5 °C; sh25, 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. Figures 15A-15B show the size exclusion chromatography (SEC) results for the main peak and high molecular weight (HMW) species of the ARGX-113 formulation described in Example 4, respectively. sh5, shaking at 5 °C; sh25, 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. FIGS. 16A-16B show the chemical degradation measured by iCE for the major peak, acidic variant, and basic variant of the ARGX-113 formulation described in Example 4, respectively. sh5, shaking at 5 °C; sh25, 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. FIGS. 17A-17D show subvisible particles of the ARGX-113 formulation described in Example 4, with diameters ≥2 μm, ≥5 μm, ≥10 μm, and ≥25 μm, respectively. sh5, shaking at 5 °C; sh25, 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. Specific details for implementing the invention

[0050] ARGX -113

[0051] In some embodiments, the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ No. 1, SEQ No. 2, or SEQ No. 3. ARGX-113 is a variant Fc site of human IgG, wherein the Fc site comprises the Y, T, E, K, F, and Y amino acids at EU positions 52, 254, 256, 433, 434, and 436, respectively.

[0052] In particular, ARGX-113 is an isolated FcRn antagonist composed of a variant Fc site, and the aforementioned variant Fc site is composed of an Fc domain homomer, wherein the amino acid sequence of each Fc domain is composed of sequence number 1.

[0053]

[0054] The N-terminal aspartic acid residue (D) in sequence number 1 corresponds to EU position 221, and the C-terminal lysine (K) in sequence number 2 corresponds to EU position 447.

[0055] In some embodiments, the water-soluble formulation and pharmaceutical composition of the present invention is a significantly homogeneous polypeptide of SEQ ID NO. 1. In some embodiments, the water-soluble formulation and pharmaceutical composition of the present invention comprises a polypeptide group, wherein at least 90% of the polypeptide (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) consists of Fc domain homomers, wherein the amino acid sequence of each Fc domain of the homomer consists of SEQ ID NO. 1.

[0056] In some embodiments, the water-soluble formulation and pharmaceutical composition of the present invention is a significantly homogeneous polypeptide of SEQ ID NO. 2. In a particularly preferred embodiment, the water-soluble formulation and pharmaceutical composition of the present invention comprises a polypeptide group wherein at least 90% of the polypeptide (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) consists of Fc domain homomers, wherein the amino acid sequence of each Fc domain of the homomer consists of SEQ ID NO. 2. In some embodiments, 90% of the polypeptide lacks a C-terminal lysine (K) residue at EU position 448.

[0057] In some embodiments, each Fc domain of ARGX-113 further comprises an N-linked glycan at EU position 297, wherein the N-linked glycan is bisecting N -Acetylglucosamine ( N It has the structure of acetylglucosamine (GlcNAc).

[0058] In some embodiments, the water-soluble formulation and pharmaceutical composition of the present invention is a significantly homogeneous polypeptide of SEQ ID NO. 3. In some embodiments, the water-soluble formulation and pharmaceutical composition of the present invention comprises a polypeptide group wherein at least 90% of the polypeptide (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) consists of Fc domain homomers, wherein the amino acid sequence of each Fc domain of the homomer consists of SEQ ID NO. 3.

[0059] The formulations and compositions of the present invention will find use in the treatment of antibody-mediated and / or antibody-associated autoimmune diseases.

[0060] Antibody-mediated autoimmune disease

[0061] The formulations and compositions of the present invention will find use in the treatment of antibody-mediated and / or antibody-associated autoimmune diseases.

[0062] Antibody-mediated and / or antibody-associated autoimmune diseases are well known. Examples of non-restrictive antibody-mediated and / or antibody-associated autoimmune diseases include allogenic islet graft rejection, alopecia areata, ankylosing spondylitis, antiphospholipid syndrome, autoimmune Addison's disease, Alzheimer's disease, antineutrophil cytoplasmic antibodies (ANCA), autoimmune diseases of the adrenal gland, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune myocarditis, autoimmune neutropenia, autoimmune oophoritis and orchitis, and autoimmune thrombocytopenia. (autoimmune thrombocytopenia), autoimmune urticaria, Behcet's disease, bullous pemphigoid, cardiomyopathy, Castleman's syndrome, celiac sprue-dermatitis, chronic fatigue immune dysfunction syndrome, chronic inflammatory demyelinating polyneuropathy,CIDP), Churg-Strauss syndrome, cicatricial pemphigoid, CREST syndrome (calcinosis, Raynaud phenomenon, esophageal dysmotility, sclerodactyly, and telangiectasia), cold agglutinin disease, Crohn's disease, dermatomyositis, discoid lupus, essential mixed cryoglobulinemia, factor VIII deficiency, fibromyalgia-fibromyositis, glomerulonephritis, Grave's disease, Guillain-Barré syndrome, Goodpasture's syndrome, graft-versus-host disease (GVHD), Hashimoto's thyroiditis, hemophilia A, idiopathic pulmonary fibrosis, idiopathic thrombocytopenic purpura (immune thrombocytopenia (ITP)), IgA neuropathy, IgM polyneuropathies, immune-mediated thrombocytopenia, juvenile arthritis,Kawasaki's disease, lichen planus, lupus erythematosus, Meniere's disease, mixed connective tissue disease, multiple sclerosis, type 1 diabetes mellitus, multifocal motor neuropathy (MMN), myasthenia gravis (MG), paraneoplastic bullous pemphigoid, pemphigus vulgaris, pemphigus foliaceus, pernicious anemia, polyarteritis nodosa, polychrondritis, polyglandular syndromes, polymyalgia rheumatica), polymyositis and dermatomyositis, primary agammaglobulinemia, primary biliary cirrhosis, psoriasis, psoriatic arthritis, Raynaud's phenomenon, Reiter's syndrome, rheumatoid arthritis, sarcoidosis, scleroderma, Sjogren's syndrome, solid organ transplant rejection,Stiff-Man syndrome, systemic lupus erythematosus (SLE), Takayasu arteritis, toxic epidermal necrolysis (TEN), Stevens-Johnson syndrome (SJS), temporal arteritis / giant cell arteritis, thrombotic thrombocytopenia purpura, ulcerative colitis, uveitis, dermatitis herpetiformis vasculitis, anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitides, vitiligo, and Wegener's granulomatosis granulomatosis) is included.

[0063] In some embodiments, the antibody-mediated autoimmune disease is selected from the group consisting of immune thrombocytopenia (ITP) and myasthenia gravis (MG).

[0064] Formulation ( Formulas )

[0065] The formulations and compositions of the present invention will find use in any disease or condition where it is desirable to reduce serum levels of Fc-containing agents in an individual. Fc-containing agents include, without limitation, autoantibodies, therapeutic antibodies, diagnostic antibodies, and immune complexes. Additional non-limiting examples of Fc-containing agents include imaging agents (e.g., labeled antibodies), antibody-drug conjugates (ADCs), Fc fusion proteins (e.g., immunoadhesins), and immunogenic agents (e.g., non-human antibodies).

[0066] Additionally, in diseases or conditions requiring the administration of a therapeutic agent, the individual will frequently develop antibodies against the therapeutic agent (e.g., anti-drug antibodies), which, therefore, interfere with the use of the therapeutic agent for its intended therapeutic purpose or cause adverse reactions in the individual. Accordingly, the formulations and compositions disclosed herein may also be used to remove antibodies against the therapeutic agent (e.g., anti-drug antibodies) developed in the individual.

[0067] One aspect of the present invention is a water-soluble formulation comprising about 100-300 mg / mL of isolated neonatal Fc receptor (FcRn) antagonist in 20-60 mM histidine / 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, 0-15 mM L-methionine, and pH 6.0-6.5, wherein the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is SEQ ID NO. 1, It consists of sequence number 2, or sequence number 3.

[0068] In some embodiments, the water-soluble formulation comprises about 100-300 mg / mL of isolated neonatal Fc receptor (FcRn) antagonist in 20-60 mM histidine / histidine HCl, 0-70 mM sucrose, 100 mM NaCl, 0%-0.05% (w / v) polysorbate 20 (polymerase 20) or polysorbate 80 (polymerase 80), 0-15 mM L-methionine, and pH 6.0-6.5, wherein the isolated neonatal FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0069] In some embodiments, the water-soluble formulation comprises 20-60 mM histidine HCl, 0-70 mM sucrose, 100-250 mM arginine HCl, 0%-0.05% (w / v) polysorbate 20 or polysorbate 80, 0-15 mM L-methionine, and about 100-300 mg / mL of isolated neonatal Fc receptor (FcRn) antagonist at pH 6.0-6.5, wherein the isolated neonatal FcRn antagonist consists of a variant Fc site, the said variant Fc site consists of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. It consists of 3.

[0070] In some embodiments, the water-soluble formulation comprises about 100-300 mg / mL of isolated neonatal Fc receptor (FcRn) antagonist in 20-60 mM histidine HCl, 100-250 mM arginine HCl, 0%-0.05% (w / v) polysorbate 20 (polymerase 20) or polysorbate 80 (polymerase 80), 0-15 mM L-methionine, and pH 6.0-6.5, wherein the isolated neonatal FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0071] In each aspect and embodiment mentioned above, in some embodiments, the water-soluble formulation contains 20 or 50 mM histidine.

[0072] In each aspect and embodiment mentioned above, in some embodiments, the water-soluble formulation comprises 0.02%-0.04 (w / v) polysorbate 20 or polysorbate 80.

[0073] In each aspect and embodiment mentioned above, in some embodiments, the water-soluble formulation contains 0 or 10 mM L-methionine.

[0074] In each of the aforementioned aspects and embodiments, in some embodiments, the pH is 6.0 or 6.5.

[0075] In each aspect and embodiment mentioned above, in some embodiments, the amino acid sequence of each Fc domain consists of sequence number 1.

[0076] One aspect of the present invention is a water-soluble formulation comprising about 100 to about 300 mg / mL of isolated neonatal FcRn antagonist in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.02%-0.04% (w / v) polysorbate 20 or polysorbate 80, pH 6.0, wherein the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0077] One aspect of the present invention is a water-soluble formulation comprising about 100 to about 200 mg / mL of isolated FcRn antagonist in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.02%-0.04% (w / v) polysorbate 20 or polysorbate 80, pH 6.0, wherein the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0078] One aspect of the present invention is a water-soluble formulation comprising about 100-300 mg / mL of isolated neonatal Fc receptor (FcRn) antagonist in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.02%-0.04% (w / v) polysorbate 20 or polysorbate 80, pH 6.0, wherein the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0079] In one embodiment, the present invention is a water-soluble formulation comprising about 100 to about 300 mg / mL of ARGX-113 in 20 mM histidine / histidineHCl, 60 mM sucrose, 100 mM NaCl, and 0.02%-0.04% (w / v) polysorbate 20 or polysorbate 80, pH 6.0, wherein ARGX-113 is an isolated FcRn antagonist composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1.

[0080] One aspect of the present invention is a water-soluble formulation comprising about 100-200 mg / mL of isolated neonatal Fc receptor (FcRn) antagonist in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.02%-0.04% (w / v) polysorbate 20 or polysorbate 80, pH 6.0, wherein the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0081] In one embodiment, the present invention is a water-soluble formulation comprising about 100 to about 200 mg / mL of ARGX-113 in 20 mM histidine / histidineHCl, 60 mM sucrose, 100 mM NaCl, and 0.02%-0.04% (w / v) polysorbate 20 or polysorbate 80, pH 6.0, wherein ARGX-113 is an isolated FcRn antagonist composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1.

[0082] As used herein, the term "about" means an amount within ± 10% of any given specific amount. For example, about 200 mg / mL includes 90% to 110% of 200 mg / mL, i.e., 180 to 220 mg / mL.

[0083] In some embodiments, the water-soluble formulation contains 100-300 mg / mL ARGX-113.

[0084] In some embodiments, the water-soluble formulation contains 100-200 mg / mL ARGX-113.

[0085] In some embodiments, the water-soluble formulation contains ARGX-113 in an amount of about 150 to about 200 mg / mL. In some embodiments, the water-soluble formulation contains about 150-200 mg / mL of ARGX-113.

[0086] In some embodiments, the water-soluble formulation contains ARGX-113 in amounts of about 150 to about 180 mg / mL. In some embodiments, the water-soluble formulation contains about 150-180 mg / mL of ARGX-113.

[0087] In some embodiments, the water-soluble formulation contains about 165 mg / mL of ARGX-113.

[0088] In some embodiments, the water-soluble formulation contains about 175 mg / mL of ARGX-113.

[0089] In some embodiments, the water-soluble formulation contains about 180 mg / mL of ARGX-113.

[0090] In some embodiments, the water-soluble formulation contains about 250 mg / mL of ARGX-113.

[0091] In some embodiments, the water-soluble formulation contains about 300 mg / mL of ARGX-113.

[0092] According to each of the previously mentioned embodiments, in some embodiments, the water-soluble formulation comprises 0.02%-0.04% (w / v) polysorbate 20. In some embodiments, the water-soluble formulation comprises 0.02% (w / v) polysorbate 20. In some embodiments, the water-soluble formulation comprises 0.03% (w / v) polysorbate 20. In some embodiments, the water-soluble formulation comprises 0.04% (w / v) polysorbate 20.

[0093] Otherwise, according to each of the previously mentioned embodiments, in some embodiments, the water-soluble formulation comprises 0.02%–0.04% (w / v) polysorbate 80. In some embodiments, the water-soluble formulation comprises 0.02% (w / v) polysorbate 80. In some embodiments, the water-soluble formulation comprises 0.03% (w / v) polysorbate 80. In some embodiments, the water-soluble formulation comprises 0.04% (w / v) polysorbate 80.

[0094] In one embodiment, the water-soluble formulation comprises 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, wherein ARGX-113 is an isolated FcRn antagonist composed of a variant Fc site, the aforementioned variant Fc site consists of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1.

[0095] One aspect of the present invention is a water-soluble formulation comprising about 150 mg / mL of isolated FcRn antagonist in 20 mM histidine / histidineHCl, 60 mM sucrose, 100 mM NaCl, and 0.04% (w / v) polysorbate 20, pH 6.0, wherein the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0096] In one embodiment, the invention is a water-soluble formulation comprising 150 mg / mL of ARGX-113 in 20 mM histidine / histidineHCl, 60 mM 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 site, the aforementioned variant Fc site consists of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0097] One aspect of the present invention is a water-soluble formulation comprising about 175 mg / mL of isolated FcRn antagonist in 20 mM histidine / histidineHCl, 60 mM sucrose, 100 mM NaCl, and 0.04% (w / v) polysorbate 20, pH 6.0, wherein the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0098] In one embodiment, the present invention is a water-soluble formulation comprising 175 mg / mL of ARGX-113 in 20 mM histidine / histidineHCl, 60 mM sucrose, 100 mM NaCl, and 0.04% (w / v) polysorbate 20, pH 6.0, wherein ARGX-113 is an isolated FcRn antagonist composed of a variant Fc site, the aforementioned variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1.

[0099] One aspect of the present invention is a water-soluble formulation comprising about 200 mg / mL of isolated FcRn antagonist in 20 mM histidine / histidineHCl, 60 mM sucrose, 100 mM NaCl, and 0.04% (w / v) polysorbate 20, pH 6.0, wherein the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0100] In one embodiment, the present invention is a water-soluble formulation comprising 200 mg / mL of ARGX-113 in 20 mM histidine / histidineHCl, 60 mM 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 site, the aforementioned variant Fc site consists of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0101] One aspect of the present invention is a water-soluble formulation comprising about 100-300 mg / mL of 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 or polysorbate 80, pH 6.0, wherein the isolated FcRn antagonist is composed of a variant Fc site, wherein the said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0102] In one embodiment, the present invention is a water-soluble formulation comprising 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%-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 site, the aforementioned variant Fc site consists of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0103] One aspect of the present invention is a water-soluble formulation comprising about 100-200 mg / mL of isolated FcRn antagonist in 20 mM histidine / 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 the isolated FcRn antagonist is composed of a variant Fc site, said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0104] In one embodiment, the present invention is a water-soluble formulation comprising about 100-200 mg / mL of ARGX-113 in 20 mM histidine / 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 ARGX-113 is an isolated FcRn antagonist composed of a variant Fc site, the aforementioned variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1.

[0105] One aspect of the present invention is a water-soluble formulation comprising about 165 mg / mL of isolated neonatal Fc receptor (FcRn) antagonist in 20 mM histidine / histidineHCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.04% (w / v) polysorbate 20, pH 6.0, wherein the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0106] In one embodiment, the present invention is a water-soluble formulation comprising 165 mg / mL of ARGX-113 in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.04% (w / v) polysorbate 20, pH 6.0, wherein ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc site, the aforementioned variant Fc site consists of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0107] One aspect of the present invention is a water-soluble formulation comprising about 175 mg / mL of isolated FcRn antagonist in 20 mM histidine / histidineHCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.03% (w / v) polysorbate 20, pH 6.0, wherein the isolated FcRn antagonist is composed of a variant Fc site, said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0108] In one embodiment, the present invention is a water-soluble formulation comprising 175 mg / mL of ARGX-113 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, wherein ARGX-113 is an isolated FcRn antagonist composed of a variant Fc site, the aforementioned variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1.

[0109] One aspect of the present invention is a water-soluble formulation comprising about 200 mg / mL of isolated FcRn antagonist in 20 mM histidine / histidineHCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.03% (w / v) polysorbate 20, pH 6.0, wherein the isolated FcRn antagonist is composed of a variant Fc site, said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0110] In one embodiment, the present invention is a water-soluble formulation comprising 200 mg / mL of ARGX-113 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, wherein ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc site, the aforementioned variant Fc site consists of an Fc domain homomer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0111] One aspect of the present invention is a water-soluble formulation comprising about 100 to about 300 mg / mL of isolated FcRn antagonist in 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 is composed of a variant Fc site, wherein the said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0112] In one embodiment, the present invention is a water-soluble formulation comprising about 100 to about 300 mg / mL of ARGX-113 in 50 mM histidine / histidine HCl, 60 mM sucrose, 150 mM arginine HCl, and 0.02%-0.04% (w / v) polysorbate 20 or polysorbate 80 at pH 6.0, wherein ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc site, the said variant Fc site consists of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0113] In some embodiments according to aspects of the present invention, the water-soluble formulation comprises 100-300 mg / mL of ARGX-113.

[0114] In some embodiment according to an aspect of the present invention, the water-soluble formulation comprises ARGX-113 in an amount of about 150 to about 200 mg / mL. In some embodiment according to an aspect of the present invention, the water-soluble formulation comprises ARGX-113 in an amount of 150-200 mg / mL.

[0115] In some embodiment according to an aspect of the present invention, the water-soluble formulation comprises ARGX-113 in an amount of about 150 to about 180 mg / mL. In some embodiment according to an aspect of the present invention, the water-soluble formulation comprises ARGX-113 in an amount of 150-180 mg / mL.

[0116] According to each of the previously mentioned embodiments, in some embodiments, the water-soluble formulation comprises 0.02%-0.04% (w / v) polysorbate 20. In some embodiments, the water-soluble formulation comprises 0.02% (w / v) polysorbate 20. In some embodiments, the water-soluble formulation comprises 0.03% (w / v) polysorbate 20. In some embodiments, the water-soluble formulation comprises 0.04% (w / v) polysorbate 20.

[0117] One aspect of the present invention is a water-soluble formulation comprising about 100 to about 200 mg / mL of isolated FcRn antagonist in 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 is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0118] In one embodiment, the present invention is a water-soluble formulation comprising about 100 to about 200 mg / mL of ARGX-113 in 50 mM histidine / histidine HCl, 60 mM sucrose, 150 mM arginine HCl, and 0.02%-0.04% (w / v) polysorbate 20 or polysorbate 80 at pH 6.0, wherein ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc site, the said variant Fc site consists of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0119] In some embodiments according to aspects of the present invention, the water-soluble formulation comprises 100-300 mg / mL of ARGX-113.

[0120] In some embodiment according to an aspect of the present invention, the water-soluble formulation comprises ARGX-113 in an amount of about 150 to about 200 mg / mL. In some embodiment according to an aspect of the present invention, the water-soluble formulation comprises ARGX-113 in an amount of 150-200 mg / mL.

[0121] In some embodiment according to an aspect of the present invention, the water-soluble formulation comprises ARGX-113 in an amount of about 150 to about 180 mg / mL. In some embodiment according to an aspect of the present invention, the water-soluble formulation comprises ARGX-113 in an amount of 150-180 mg / mL.

[0122] In one embodiment, the water-soluble formulation contains about 150 mg / mL ARGX-113 in 50 mM histidine / histidine HCl, 60 mM sucrose, 150 mM arginine HCl, and 0.04% (w / v) polysorbate 80, pH 6.0.

[0123] In one embodiment, the water-soluble formulation contains about 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) polysorbate 80, pH 6.0.

[0124] One aspect of the present invention is a water-soluble formulation comprising about 100-200 mg / mL of isolated FcRn antagonist in 20 mM histidine / histidine 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 the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0125] In one embodiment, the present invention is a water-soluble formulation comprising about 100-200 mg / mL of ARGX-113 in 20 mM histidine / histidine 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-113 is an isolated FcRn antagonist composed of a variant Fc site, the aforementioned variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1.

[0126] One aspect of the present invention is a water-soluble formulation comprising about 175 mg / mL of isolated FcRn antagonist 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 antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0127] In one embodiment, the present invention is a water-soluble formulation comprising 175 mg / mL ARGX-113 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 ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc site, the aforementioned variant Fc site consists of an Fc domain homomer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0128] One aspect of the present invention is a water-soluble formulation comprising about 200 mg / mL of isolated FcRn antagonist in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM arginine HCl, 0 mM L-methionine, and 0.03% (w / v) polysorbate 20, pH 6.0, wherein the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of an Fc domain homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0129] In one embodiment, the present invention is a water-soluble formulation comprising 200 mg / mL ARGX-113 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 ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc site, the aforementioned variant Fc site consists of an Fc domain homomer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0130] One aspect of the present invention is a water-soluble formulation comprising about 100-300 mg / mL of isolated neonatal Fc receptor (FcRn) antagonist in 50 mM histidine / histidine HCl, 200 mM arginine HCl, pH 6.5, wherein the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0131] In one embodiment, the water-soluble formulation comprises about 200-300 mg / mL of isolated neonatal Fc receptor (FcRn) antagonist in 50 mM histidine / histidine HCl, 200 mM arginine HCl, pH 6.5, wherein the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1.

[0132] In one embodiment, the water-soluble formulation comprises about 250-300 mg / mL of isolated neonatal Fc receptor (FcRn) antagonist in 50 mM histidine / histidine HCl, 200 mM arginine HCl, pH 6.5, wherein the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1.

[0133] According to each prior art aspect and embodiment, in some embodiments, the water-soluble formulation is substantially free of dissolved oxygen. As used herein, "substantially free" means that at least 95% is absent. For example, in some embodiments, the water-soluble formulation is at least 95% free of dissolved oxygen. In various embodiments, the water-soluble formulation is at least 96%, at least 97%, at least 98%, at least 99%, or 100% free of dissolved oxygen. Assuming that water is generally in equilibrium with air, 20% oxygen, in some embodiments, the water-soluble formulation that is substantially free of dissolved oxygen contains less than 1% or 1% dissolved oxygen. In some embodiments, the water-soluble formulation is 100% free of dissolved oxygen.

[0134] According to each prior art aspect and embodiment, in some embodiments, the water-soluble formulation is suitable for in vivo use. For example, in such embodiments, the water-soluble formulation is sterile and does not contain pharmaceutically unacceptable amounts of toxic substances such as endotoxins. Such water-soluble formulation may meet quality standards of Good Manufacturing Process (GMP) in accordance with regulations promulgated by, for example, the US Food and Drug Administration (FDA).

[0135] Methods of making formulations

[0136] The formulation according to the present invention can be prepared using any suitable method. Generally, ARGX-113 can be prepared from an expression vector or from eukaryotic cells containing a nucleic acid sequence encoding an Fc domain. For example, eukaryotic cells include Chinese hamster ovary (CHO) cells), DG44 and DUXB11 (Chinese hamster ovary lines, DHFR-negative), HELA (human cervical carcinoma), CVI (monkey kidney line), COS (CVI derivative with SV40 T antigen), R1610 (Chinese hamster fibroblast), BALBC / 3T3 (mouse fibroblast), HAK (hamster kidney line), SP2 / 0 (mouse myeloma), BFA-1c1BPT (bovine endothelial cells), RAJI (human lymphocyte), 293 It may be a human kidney or NS0 cell. In one embodiment, the eukaryotic cell used to express ARGX-113 is a CHO cell. For example, see WO 2015 / 100299, the full text of which is incorporated herein by reference. ARGX-113 is typically expressed as a secreted protein that can be isolated from cells using techniques known in the art. Generally, the isolated and unconcentrated protein product is then placed in a sterile aqueous solution such as Tris / Glycine, pH 7.2 or 20 mM L-histidine / L-histidine HCl·H2O, pH 6.0.

[0137] This initial product is then up-concentrated and appropriately buffered to reach a concentrated protein solution containing ARGX-113 at a concentration equivalent to or greater than the target final concentration. For example, up-concentration and buffering may yield an intermediate product containing about 200 mg / mL of ARGX-113 in 20 mM L-histidine / L-histidine HCl·H2O, 100 mM NaCl, pH 6.0.

[0138] High concentration can be performed using any suitable method of the technology in this field. Such methods may include, without limitation, tangential flow filtration (TFF), dialysis, ultrafiltration, and lyophilization. For commercial production purposes, TFF may typically be used.

[0139] Additional ingredients may subsequently be added to achieve a desired final formulation. For example, additional ingredients such as NaCl, arginine HCl, sucrose, and / or polysorbate may be added from concentrated stock solutions of each of the mentioned additional ingredients, and if necessary, water may be added to achieve a desired final formulation. In a specific embodiment, polysorbate (PS20) is added as the final additive to the formulation to achieve an accurate pH value (adding polysorbate last avoids an increase in concentration due to the molecular weight of polysorbate with ARGX-113).

[0140] In some embodiments, the intermediate solution and additional components are otherwise treated to remove gas or to reduce or remove dissolved oxygen. For example, the aforementioned intermediate solution and components may be equilibrated with argon or nitrogen.

[0141] In some embodiments, the final aqueous formulation is otherwise treated to remove gas or to reduce or remove dissolved oxygen. For example, the final aqueous formulation may be equilibrated with argon or nitrogen by bubbling the final aqueous formulation with argon or nitrogen for a sufficient time to reduce or remove dissolved oxygen from the formulation. In some embodiments, the final aqueous formulation is then stored under a nitrogen atmosphere.

[0142] The water-soluble formulation prepared in this way will typically be sterile filtered and then aliquoted and stored in a sterile container or device as described herein.

[0143] Routes of administration

[0144] The water-soluble formulation of the invention is suitable for parenteral administration. In some embodiments, the water-soluble formulation of the invention is suitable for subcutaneous administration. In some embodiments, the water-soluble formulation of the invention is suitable for intravenous administration. In some embodiments, the water-soluble formulation of the invention is suitable for intraperitoneal administration.

[0145] Effective amount

[0146] The formulation and composition are generally administered in an effective amount. "Effective amount" means an amount sufficient to achieve a desired effect. In some embodiments, the effective amount is a therapeutically effective amount, that is, an amount sufficient to achieve a desired therapeutic effect in the individual. Examples of desired therapeutic effects include, without limitation, a reduction in serum total IgG, and the treatment of various antibody-mediated autoimmune diseases such as myasthenia gravis (MG) and immune thrombocytopenia (ITP).

[0147] As used herein, "subject" generally means a mammal. In some embodiments, the subject is a non-human mammal or a non-human primate. In some embodiments, 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 human, i.e., a human who is at least 18 years of age. In some embodiments, the subject is a human who is 18 years of age or younger.

[0148] Pharmaceutical product

[0149] One aspect of the present invention is a packaged pharmaceutical product comprising a sterile container containing a therapeutically effective amount of a water-soluble formulation of the present invention. In various embodiments, the packaged pharmaceutical product may be presented as a single-use vial, a multi-use vial, or a pre-filled syringe.

[0150] Devices

[0151] One aspect of the present invention is a device comprising a therapeutically effective amount of the water-soluble formulation of the present invention.

[0152] In some embodiments, the water-soluble formulation comprises about 100-200 mg / mL of isolated FcRn antagonist in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.02%-0.04% (w / v) polysorbate 20 or polysorbate 80, pH 6.0, wherein the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0153] In some embodiments, the water-soluble formulation comprises 150 mg / mL of isolated FcRn antagonist in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.04% (w / v) polysorbate 20, pH 6.0, wherein the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of sequence number 1, sequence number 2, or sequence number 3.

[0154] In some embodiments, the water-soluble formulation comprises 175 mg / mL of isolated FcRn antagonist in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.04% (w / v) polysorbate 20, pH 6.0, wherein the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of sequence number 1, sequence number 2, or sequence number 3.

[0155] In some embodiments, the water-soluble formulation comprises 200 mg / mL of isolated FcRn antagonist in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.04% (w / v) polysorbate 20, pH 6.0, wherein the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0156] In some embodiments, the water-soluble formulation is a water-soluble formulation comprising about 100-300 mg / mL of ARGX-113 in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0%-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 site, the aforementioned variant Fc site consists of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0157] In some embodiments, the water-soluble formulation is a water-soluble formulation comprising about 100-200 mg / mL of ARGX-113 in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.02%-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 site, the aforementioned variant Fc site consists of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0158] In some embodiments, the water-soluble formulation is a water-soluble formulation comprising 150 mg / mL of ARGX-113 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 an isolated FcRn antagonist consisting of a variant Fc site, the aforementioned variant Fc site consists of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0159] In some embodiments, the water-soluble formulation is a water-soluble formulation comprising 175 mg / mL of ARGX-113 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 an isolated FcRn antagonist consisting of a variant Fc site, the aforementioned variant Fc site consists of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0160] In some embodiments, the water-soluble formulation is a water-soluble formulation comprising 200 mg / mL of ARGX-113 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 an isolated FcRn antagonist consisting of a variant Fc site, the aforementioned variant Fc site consists of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0161] In some embodiments, the water-soluble formulation comprises about 100-300 mg / mL of 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 or polysorbate 80, pH 6.0, wherein the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0162] In some embodiments, the water-soluble formulation is a water-soluble formulation comprising 150 mg / mL of isolated FcRn antagonist in 20 mM histidine / 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 antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0163] In some embodiments, the water-soluble formulation is a water-soluble formulation comprising 165 mg / mL of isolated FcRn antagonist in 20 mM histidine / 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 antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0164] In some embodiments, the water-soluble formulation is a water-soluble formulation comprising 175 mg / mL of isolated FcRn antagonist in 20 mM histidine / 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 antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0165] In some embodiments, the water-soluble formulation comprises 180 mg / mL of isolated FcRn antagonist in 20 mM histidine / 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 antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0166] In some embodiments, the water-soluble formulation comprises 200 mg / mL of isolated FcRn antagonist in 20 mM histidine / 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 antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0167] In some embodiments, the water-soluble formulation comprises 300 mg / mL of isolated FcRn antagonist in 20 mM histidine / 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 antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0168] In some embodiments, the water-soluble formulation is a water-soluble formulation comprising 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%-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 site, the aforementioned variant Fc site consists of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0169] In some embodiments, the water-soluble formulation is a water-soluble formulation comprising about 150 mg / mL of ARGX-113 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, wherein ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc site, the aforementioned variant Fc site consists of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0170] In some embodiments, the water-soluble formulation is a water-soluble formulation comprising about 165 mg / mL of ARGX-113 in 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.04% (w / v) polysorbate 20, pH 6.0, wherein ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc site, the aforementioned variant Fc site consists of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0171] In some embodiments, the water-soluble formulation comprises about 165 mg / mL of isolated FcRn antagonist in 20 mM histidine / 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 antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0172] In some embodiments, the water-soluble formulation is a water-soluble formulation comprising 175 mg / mL of ARGX-113 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, wherein ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc site, the aforementioned variant Fc site consists of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0173] In some embodiments, the water-soluble formulation is a water-soluble formulation comprising 200 mg / mL of ARGX-113 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, wherein ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc site, the aforementioned variant Fc site consists of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0174] In some embodiments, the water-soluble formulation comprises about 100 to about 200 mg / mL of isolated FcRn antagonist in 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 is composed of a variant Fc site, wherein the said variant Fc site is composed of two Fc domains forming a homodimer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0175] In some embodiments, the water-soluble formulation is a water-soluble formulation comprising about 100 to about 200 mg / mL of ARGX-113 in 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 ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc site, the aforementioned variant Fc site consists of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0176] In some embodiments, the water-soluble formulation is a water-soluble formulation comprising about 100 to about 200 mg / mL of isolated FcRn antagonist in 50 mM histidine / histidine 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 the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0177] In some embodiments, the water-soluble formulation comprises about 150 mg / mL of isolated FcRn antagonist in 50 mM histidine / histidine HCl, 60 mM sucrose, 100 mM arginine HCl, 10 mM L-methionine, and 0.02-0.04% (w / v) polysorbate 20, pH 6.0, wherein the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0178] In some embodiments, the water-soluble formulation comprises about 175 mg / mL of isolated FcRn antagonist in 50 mM histidine / histidine HCl, 60 mM sucrose, 100 mM arginine HCl, 10 mM L-methionine, and 0.02-0.04% (w / v) polysorbate 20, pH 6.0, wherein the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0179] In some embodiments, the water-soluble formulation comprises about 200 mg / mL of isolated FcRn antagonist in 50 mM histidine / histidine HCl, 60 mM sucrose, 100 mM arginine HCl, 10 mM L-methionine, and 0.02-0.04% (w / v) polysorbate 20, pH 6.0, wherein the isolated FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0180] In some embodiments, the water-soluble formulation is a water-soluble formulation comprising about 100 to about 200 mg / mL of ARGX-113 in 50 mM histidine / histidine 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-113 is an isolated FcRn antagonist consisting of a variant Fc site, the aforementioned variant Fc site consists of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0181] In some embodiments, the water-soluble formulation is a water-soluble formulation comprising about 150 mg / mL of ARGX-113 in 50 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 ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc site, the aforementioned variant Fc site consists of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0182] In some embodiments, the water-soluble formulation is a water-soluble formulation comprising about 175 mg / mL of ARGX-113 in 50 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 ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc site, the aforementioned variant Fc site consists of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0183] In some embodiments, the water-soluble formulation is a water-soluble formulation comprising about 200 mg / mL of ARGX-113 in 50 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 ARGX-113 is an isolated FcRn antagonist consisting of a variant Fc site, the aforementioned variant Fc site consists of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain consists of SEQ ID NO. 1.

[0184] In one embodiment, the water-soluble formulation comprises about 100-300 mg / mL of isolated neonatal Fc receptor (FcRn) antagonist in 50 mM histidine HCl, 200 mM arginine HCl, pH 6.5, wherein the isolated neonatal FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1, SEQ ID NO. 2, or SEQ ID NO. 3.

[0185] In one embodiment, the water-soluble formulation comprises about 200-300 mg / mL of isolated neonatal Fc receptor (FcRn) antagonist in 50 mM histidine HCl, 200 mM arginine HCl, pH 6.5, wherein the isolated neonatal FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1.

[0186] In one embodiment, the water-soluble formulation comprises about 250-300 mg / mL of isolated neonatal Fc receptor (FcRn) antagonist in 50 mM histidine HCl, 200 mM arginine HCl, pH 6.5, wherein the isolated neonatal FcRn antagonist is composed of a variant Fc site, the said variant Fc site is composed of two Fc domains forming a homomer, wherein the amino acid sequence of each Fc domain is composed of SEQ ID NO. 1.

[0187] According to each preceding embodiment of this aspect, in some embodiments, the device comprises a water-soluble formulation of about 1 to about 2.5 mL. According to each preceding embodiment of this aspect, in some embodiments, the device comprises a water-soluble formulation of 1 to 2.5 mL.

[0188] In some embodiments, the device comprises a syringe containing a water-soluble formulation. Such a syringe may optionally be fitted with a needle suitable for administering at least a portion of the water-soluble solution contained within the syringe to an individual. Fine gauge needles (small diameter) should be used for drugs with low viscosity that cause less pain to the patient. The needle gauge is preferably 27 gauge or higher (i.e., smaller diameter). The outer diameter of the needle may be 0.413 mm, 0.41 mm, or smaller.

[0189] In some embodiments, the syringe is presented as a pre-filled syringe. Such a pre-filled syringe may be suitable for single use or, on the other hand, for multiple uses (two or more). Such a pre-filled syringe may optionally be fitted with a needle suitable for administering at least a portion of the aqueous solution contained in the pre-filled syringe to an individual. In some embodiments, the pre-filled syringe is presented as a single-unit package.

[0190] In some embodiments, the pre-filled syringe is largely free of ambient air. That is, in such embodiments, the water-soluble formulation contained within the pre-filled syringe is largely free of dissolved oxygen. For example, the water-soluble formulation contained within the pre-filled syringe may be prepared with nitrogen as described herein and then placed into the syringe and sealed under a nitrogen atmosphere to exclude atmospheric oxygen. In some such embodiments, the pre-filled syringe may be presented in gas-impermeable packaging.

[0191] In a particular embodiment, the present invention is a pre-filled syringe filled with a 2 mL or 2.1 mL aqueous formulation containing, for example, 360 mg / 2 mL (= 180 mg / mL) or 330 mg / 2 mL (= 165 mg / mL) of an isolated neonatal Fc receptor (FcRn) antagonist such as ARGX-113, as described herein. On the other hand, the present invention is a vial filled with a 2.2 mL aqueous formulation containing, for example, 360 mg / 2.2 mL (= 165 mg / mL) of an isolated neonatal Fc receptor (FcRn) antagonist such as ARGX-113, as described herein. Such a vial may be in a kit with a needle suitable for administering at least a portion of the aqueous solution contained in the vial to an individual.

[0192] This immediate invention further considers additional devices comprising 2 mL, 2.1 mL, 2.2 mL, or about 2.5 mL or more of a water-soluble formulation according to the present invention. Such devices may comprise, for example and without limitation, about 1.8 mL, 2 mL, 2.1 mL, 2.2 mL, 2.4 mL, 2.6 mL, 2.8 mL, 3 mL, 5 mL, about 10 mL, about 20 mL, about 50 mL, and about 100 mL of a water-soluble formulation according to the present invention. This has the advantage that the patient (male / female) can administer this formulation one shot by subcutaneous injection, for example, using a syringe pre-filled with 2 mL, 2.1 mL, 2.2 mL, or 5 mL of a water-soluble formulation according to the present invention. Such a "push" subcutaneous administration takes about 12 to 20 seconds or up to 1 minute. For comparison: Infusion into the individual by a nurse or caregiver can take from a few minutes to several hours; IV (intravenous) infusion of the ARGX-113 formulation takes about 60 minutes. Pre-filled syringes offer advantages to the patient because they can be used as a subcutaneous maintenance dose via self-administration.

[0193] The present invention is further illustrated by the following embodiments, which should not be construed as further limiting. The contents of the Sequence Listing, figures, and all references, patents, and published patent applications cited in this application are specifically incorporated herein by reference.

[0194] Example ( EXAMPLES)

[0195] Example 1. Rheological Characterization

[0196] This example describes experiments performed to develop high-concentration ARGX-113 formulations and characterize their properties. In particular, the objective of this set of experiments was to identify the concentration-viscosity correlation. The study was conducted at two temperatures: 5 °C and 25 °C. To identify the rheological profile of ARGX-113, five concentrations of ARGX-113 were screened in platform buffer at optimal shear rates.

[0197] Starting with 4–5 mg / mL of ARGX-113 in Tris / glycine buffer pH 7.2, ARGX-113 was exchanged and highly concentrated to the maximum possible concentration (targeting 250 mg / mL) in sodium phosphate pH 6.7 + Salt and separately in histidine HCl (HisHCl) + Salt pH 6.0. Concentration and pH were monitored during this process. The viscoelastic reaction was observed during shear rate ramping from 0–4000 s. -1 HisHCl was studied at the highest concentration in pH 6.0 + salt. Serial dilutions (6 concentrations) were performed and UV-absorbance measurements (A 280 It was proven by ). The concentration-to-viscosity measurement was at a shear rate of 2000s. -1 It was performed in.

[0198] ARGX-113 in sodium phosphate pH 6.7 + salt was precipitated from the solution at ~130 mg / mL.

[0199] The up-concentration processing of ARGX-113 in sodium phosphate pH 6.7+ salt is very slow. High concentrations (~130 mg / mL) of ARGX-113 in sodium phosphate pH 6.7+ salt exhibited a reversible solid / liquid phase transition depending on storage conditions (25 °C versus 5 °C). High concentrations (~130 mg / mL) of ARGX-113 in sodium phosphate pH 6.7+ salt showed visible particles of very high numbers and sizes at 5 °C.

[0200] In contrast, ARGX-113 in HisHCl + salt (Salt) pH 6.0 remained as a solution up to at least ~260 mg / mL.

[0201] The ARGX-113 high-concentration process was fast in HisHCl + salt (Salt) pH 6.0.

[0202] In contrast to sodium phosphate buffer + salt, no significant precipitation or phase separation was observed during high concentration.

[0203] After storage at 5 °C, a concentration-dependent sol-gel transition was observed. The HisHCl formulation appeared very jelly-like at a concentration of ~260 mg / mL (260 > 200 >> 120 mg / mL) and turned into a liquid when warmed to room temperature or pipetted.

[0204] The shear thinning / concentration behavior of ARGX-113 at the highest concentration (HisHCl + salt, pH 6.0, 267 mg / mL, 25 °C) is 0–4000 s as the shear rate increases -1로 연구되었다 Typical results are shown in Fig. 1. As shown in the figure, ARGX-113 runs for 1,000–4,000 s-1 No significant shear thickening or exfoliation was observed within the range. For further study, the 2000s -1 This shear rate was selected.

[0205] Representative results of the study on the viscosity versus concentration of ARGX-113 at HisHCl + salt, pH 6.0 are shown in Figure 2. As shown in the figure, the viscosity at approximately 180 mg / mL at 5 °C and 25 °C was 6 and 17 mPa·s, respectively, and the viscosity at approximately 200 mg / mL at 5 °C and 25 °C was 9 and 33 mPa·s, respectively.

[0206] Starting with 4-5 mg / mL of ARGX-113 in Tris / glycine buffer pH 7.2, ARGX-113 was exchanged and highly concentrated to a target concentration of 175 mg / mL in sodium phosphate + salt, pH 6.7, and separately in HisHCl + salt, pH 6.0. Concentration and pH were monitored during this process. Stock solutions of different additives were prepared to obtain various target formulation compositions. Eleven (11) formulation conditions with different additives and pH were studied to evaluate viscosity reduction at high concentrations of 175 mg / mL. The various formulations studied are shown in Table 1.

[0207] ID buffer pH Additive 1 Additive 2 Additive 3 F1 20 mM HisHCl 6.0 150 mM NaCl -- -- F2 50 mM HisHCl 6.0 150 mM ArgHCl -- -- F3 20 mM HisHCl 6.0 100 mM NaCl 50 mM ArgHCl -- F4 20 mM HisHCl 6.0 50 mM NaCl 50 mM ArgHCl 75 mM Sucrose F5 20 mM HisHCl 6.0 50 mM NaCl 100 mM ArgHCl -- F6 20 mM HisHCl 6.0 100 mM NaCl 75 mM Sucrose -- F7 20 mM HisHCl 6.0 50 mM NaCl 50 mM ArgHCl -- F8 20 mM HisHCl 6.0 50 mM NaCl 150 mM Sucrose -- F9 20 mM HisHCl 6.5 150 mM NaCl -- -- F10 20 mM HisHCl 5.5 150 mM NaCl -- -- F11 25 mM SodPhos 6.7 100 mM NaCl 150 mM ArgHCl --

[0208] ArgHCl: Arginine HCl

[0209] SodPhos: Sodium phosphate

[0210] Phosphate formulations could not be formulated at concentrations above 100 mg / mL due to significant precipitation. All formulations were stored at 5 °C for 48 hours to observe phase transitions, if any. Viscosity measurements were performed at a shear rate of 2000 s² for all 11 formulations. -1 It was performed at 5 °C, and optional formulations were performed at 25 °C.

[0211] Except for storage, all formulations remained liquid and clear even after 2 days of storage at 5 °C. F11 (SodPhos + NaCl) became turbid and showed precipitation at high concentrations of ~129 mg / mL. The phosphate formulations formed a clear solution after preparation and storage at 5 °C. The F9 (pH 6.5) formulation was slightly opaque and subsequently became slightly clear at 5 °C.

[0212] In all formulations F1-F11, viscosity was low at 5 °C (< 25 mPa·s). F2 and F5 showed effectively low viscosity. Sucrose increased viscosity at 175 mg / mL in 20 mM His / HisHCl, pH 6.0 (F6 and F8).

[0213] Example 2. Determination of additional rheological properties

[0214] This example describes experiments conducted to develop and characterize additional candidate high-concentration formulations of ARGX-113. In particular, the objective of this set of experiments is to identify candidate high-concentration liquid formulations of ARGX-113 for preclinical toxicology and early phase clinical studies based on specific properties and short-term stability studies.

[0215] The compositions of the four water-soluble formulations of ARGX-113 studied in this example are shown in Table 2.

[0216] ARGX-113 buffer pH Additive 1 Additive 2 Additive 3 150 mg / mL 50 mM HisHCl 6.0 -- 150 mM ArgHCl 0.04% w / v PS80 150 mg / mL 20 mM HisHCl 6.0 100 mM NaCl 60 mM Sucrose 0.04% w / v PS20 100 mg / mL 20 mM HisHCl 6.5 75 mM NaCl 100 mM Sucrose 0.04% w / v PS20 100 mg / mL 20 mM HisHCl 6.0 75 mM NaCl 100 mM Sucrose 0.04% w / v PS20

[0217] PS20: Polysorbate 20

[0218] PS80: Polysorbate 80

[0219] Starting with ARGX-113 ~136 mg / mL in 20 mM L-histidine / L-histidine HCl·H2O, ARGX-113 was exchanged to achieve target buffer concentration and pH, and then highly concentrated above the target concentration shown in Table 2. F14 was prepared at 100 mg / mL because the material solidifies during high concentration, which limited the formulation concentration to 100 mg / mL after preparation using the stock solution.

[0220] The determination of the initial properties of these formulations includes pH determination, osmotic pressure due to freezing point depression, and actual protein concentration. Representative results are shown in Table 3.

[0221] Test Formulation F12 F13 F14 F15 pH 6.0 6.1 6.4 6.0 Osmotic pressure (mOsmol / kg) 367 322 385 395 Protein concentration (mg / mL) 160.1 153.4 99.2 101.7

[0222] An aliquot of each formulation was subjected to shake stress in a horizontal position on an orbital shaker at a target speed of 115 rpm under room temperature and cold temperature conditions for approximately 7 days.

[0223] The aliquots of each formulation were subjected to five freeze / thaw cycles from -65 °C or lower in the vertical position to room temperature.

[0224] Fig. 3 As shown in [the example], ARGX-113 concentration was stable under all tested conditions. Fig. 3 From: F1 = F12; F2 = F13; F3= F14; F4 = F15. Fig. 3The bars shown from left to right are as follows: F1 initial; F1 shaking at 5°C (shaking sh 5); F1 shaking at 25°C (shaking sh 25); F1 of five cycles of freeze / thaw stress (freeze / thawF / T); F1 for 4 weeks at 5°C (4W5); F1 for 4 weeks at 25°C (4W25); F1 for 4 weeks at 40°C (4W40); F1 for 8 weeks at 5°C (8w5); F1 for 8 weeks at 25°C (8w25); F1 for 8 weeks at 40°C (8w40); F2 initial; F2 shaking at 5°C (shaking sh 5); F2 shaking at 25°C (shaking sh 25); F2 of five cycles of freeze / thaw stress (freeze / thawF / T); 4 weeks at 5°C (4W5) F2; 4 weeks at 25°C (4W25) F2; 4 weeks at 40°C (4W40) F2; 8 weeks at 5°C (8w5) F2; 8 weeks at 25°C (8w25) F2; 8 weeks at 40°C (8w40) F2; F3 initial (F3 initial); F3 shaking at 5°C (shaking sh 5); F3 shaking at 25°C (shaking sh 25); F3 of five cycles of freeze / thaw stress (freeze / thawF / T); 4 weeks at 5°C (4W5) F3; 4 weeks at 25°C (4W25) F3; 4 weeks at 40°C (4W40) F3; 8 weeks at 5°C (8w5) F3; 8 weeks at 25°C (8w25) F3; 8 weeks at 40°C (8w40) F3; F4 initial; shaking at 5°C (shaking sh 5); shaking at 25°C (shaking sh 25); F4 of five cycles of freeze / thaw stress (freeze / thawF / T); 4 weeks at 5°C (4W5) F4; 4 weeks at 25°C (4W25) F4; 4 weeks at 40°C (4W40) F4; 8 weeks at 5°C (8w5) F4;8 weeks at 25°C (8w25) F4; 8 weeks at 40°C (8w40) F4.;

[0225] Figs. 4A-4B As shown in [figure], no significant differences in aggregation and fragmentation by size exclusion chromatography (SEC) were observed in all tested liquid formulations. In addition, Figs. 5A-5C As shown in [the example], no major chemical degradation was observed in integrated chip-based capillary electrophoresis (iCE). Figs. 6A-6D As shown in [figure], the overall visible and subvisible particle counts were low under shaking and freeze / thaw stress. This data suggests that there is no significant difference between the polysorbate 20 and 80 formulations, and between both polysorbate 20 and polysorbate 80 at 0.04% w / v, and between ARGX-113, which was equally protected against shaking and freeze / thaw stress. In Figs. 4A-4B, 5A-5C and 6A-6D: F1 =F12; F2 = F13; F3=F14; F4 = F15. Figs. 4A-4B, 5A-5C and 6A-6DThe bars shown in order from left to right are as follows: F1 initial; F1 shaking at 5°C (shaking sh 5); F1 shaking at 25°C (shaking sh 25); F1 of five cycles of freeze / thaw stress (freeze / thawF / T); 4 weeks at 5°C (4W5) F1; 4 weeks at 25°C (4W25) F1; 4 weeks at 40°C (4W40) F1; 8 weeks at 5°C (8w5) F1; 8 weeks at 25°C (8w25) F1; 8 weeks at 40°C (8w40) F1; F2 initial; F2 shaking at 5°C (shaking sh 5); F2 shaking at 25°C (shaking sh 25); F2 of five cycles of freeze / thaw stress (freeze / thawF / T); 4 weeks at 5°C (4W5) F2; 4 weeks at 25°C (4W25) F2; 4 weeks at 40°C (4W40) F2; 8 weeks at 5°C (8w5) F2; 8 weeks at 25°C (8w25) F2; 8 weeks at 40°C (8w40) F2; F3 initial (F3 initial); F3 shaking at 5°C (shaking sh 5); F3 shaking at 25°C (shaking sh 25); F3 of five cycles of freeze / thaw stress (freeze / thawF / T); 4 weeks at 5°C (4W5) F3; 4 weeks at 25°C (4W25) F3; 4 weeks at 40°C (4W40) F3; 8 weeks at 5°C (8w5) F3; 8 weeks at 25°C (8w25) F3; 8 weeks at 40°C (8w40) F3; F4 initial; shaking at 5°C (shaking sh 5); shaking at 25°C (shaking sh 25); F4 of five cycles of freeze / thaw stress (freeze / thawF / T); 4 weeks at 5°C (4W5) F4; 4 weeks at 25°C (4W25) F4; 4 weeks at 40°C (4W40) F4; 8 weeks at 5°C (8w5) F4;8 weeks at 25°C (8w25) F4; 8 weeks at 40°C (8w40) F4.;

[0226] Short-term stability data up to 2 months (8 weeks) indicate that ARGX-113 exhibited a tendency toward severe aggregation with an area increase of 0.8–1.7% after 8 weeks at 40 °C SEC. The aggregation rate of ARGX-113 depended on concentration, pH, and composition under specific storage conditions. The F13 formulation containing NaCl showed higher aggregation at 40 °C compared to F12 containing arginine. F14 at pH 6.5 showed higher aggregation at a concentration of 100 mg / mL at 40 °C compared to F15 at pH 6.0. These data suggest that ARGX-113 has better stability at pH 6.0 compared to pH 6.5. The aggregation was below the limit of quantification (LOQ) in SEC.

[0227] In iCE, ARGX-113 initially exhibited highly basic and acidic species with low major peaks, and stability decreased rapidly depending on pH and composition, particularly at elevated temperatures. The iCE profile suggests that in the liquid formulation, the major peak of ARGX-113 transitioned primarily to acidic variants (~24%) during 8 weeks of stability at 40 °C. ARGX-113 at 100 mg / mL and 150 mg / mL did not show major differences in the rate and extent of chemical degradation in the liquid formulation during 2 months of stability. No major changes in the basic peak were observed across all studied formulations and stability time points.

[0228] ARGX-113 did not show significant differences in the rate and extent of chemical degradation in 2-month stability in liquid formulations at 100 mg / mL and 150 mg / mL. No major changes in the basic peak were observed across all studied formulations and stability time points.

[0229] In addition, CE-SDS (Caliper, Perimeter) did not show any major changes in the 8-week stability study (results not shown).

[0230] Except for F12, which showed many particles (particle cloud) at 40 °C after 8 weeks, all formulations showed no visible particles at 5, 25, and 40 °C initially and after 8 weeks. The color of the solution varied from slightly brown to brown depending on storage and composition conditions, and all formulations showed a stable target pH (± 0.2) at 5, 25, and 40 °C initially and after 8 weeks.

[0231] Fig. 7 As shown in [Figure], the overall turbidity of the formulation at 150 mg / mL was higher than that of 15 FNU (formazin nephelometric turbidity units). Additionally, the sodium chloride formulation exhibited higher turbidity compared to the arginine-containing formulation. F13 showed a notable increase in turbidity at 40 °C after 4 and 8 weeks. In Fig. 7: F1 = F12; F2 = F13; F3=F14; F4 = F15. Fig. 7The bars shown in order from left to right are as follows: F1 initial; F1 shaking at 5°C (shaking sh 5); F1 shaking at 25°C (shaking sh 25); F1 of five cycles of freeze / thaw stress (freeze / thawF / T); F1 for 4 weeks at 5°C (4W5); F1 for 4 weeks at 25°C (4W25); F1 for 4 weeks at 40°C (4W40); F1 for 8 weeks at 5°C (8w5); F1 for 8 weeks at 25°C (8w25); F2 initial; F2 shaking at 5°C (shaking sh 5); F2 shaking at 25°C (shaking sh 25); F2 of five cycles of freeze / thaw stress (freeze / thawF / T); 4 weeks at 5°C (4W5) F2; 4 weeks at 25°C (4W25) F2; 4 weeks at 40°C (4W40) F2; 8 weeks at 5°C (8W5) F2; 8 weeks at 25°C (8W25) F2; F3 initial; shaking at 5°C F3; shaking at 25°C F3 (shaking F3 25); F3 of five cycles of freeze / thaw stress (freeze / thawF / T); 4 weeks at 5°C (4W5) F3; 4 weeks at 25°C (4W25) F3; 4 weeks at 40°C (4W40) F3; 8 weeks at 5°C (8W5) F3; 8 weeks at 25°C (8W25) F3; F4 initial; F4 shaking at 5°C (shaking sh 5); F4 shaking at 25°C (shaking sh 25); F4 of five cycles of freeze / thaw stress (freeze / thawF / T); 4 weeks at 5°C (4W5) F4; 4 weeks at 25°C (4W25) F4; 4 weeks at 40°C (4W40) F4; 8 weeks at 5°C (8w5) F4; 8 weeks at 25°C (8w25) F4.

[0232] From the results obtained in this example, the following conclusions were drawn: (i) the formulation containing NaCl showed higher turbidity than the formulation containing arginine, and the increase in turbidity was higher in the NaCl formulation at a concentration of 150 mg / mL at 40°C; (ii) except for F12, which showed increased visible and invisible particles at 40°C, the overall number of subvisible particles was notable in the 8-week stability study; (iii) the initial level of aggregates was high, but the rate of increase in aggregates was moderate after 8 weeks at 40°C; (iv) the arginine formulation showed lower aggregation in the 8-week stability compared to the NaCl formulation; (v) ARGX-113 showed good physical stability at concentrations of 150 and 100 mg / mL, but the physical stability at the 100 mg / mL concentration was slightly higher compared to the 150 mg / mL formulation at pH 6.0; (vi) The initial levels of the electrical properties of the charged variants were high, and the main peak was mainly the acidic variant in all liquid formulations. The formulation at pH 6.5 showed slightly higher formation of the acidic variant compared to the formulation at pH 6.0; and (vii) Polysorbate 20 and Polysorbate 80 were equally effective in protecting ARGX-113 from stirring and freeze / thaw stress at a concentration of 0.04% w / v.

[0233] Example 3. pH and Surfactant Optimization

[0234] This example describes additional experiments to develop additional ARGX-113 candidate high-concentration formulations and to perform property characterization. In particular, the goal of this set of experiments is to identify candidate high-concentration liquid formulations of ARGX-113 for preclinical toxicology and early phase clinical studies based on specific properties and short-term stability studies.

[0235] The compositions of the seven water-soluble formulations of ARGX-113 studied in this example are shown in Table 4.

[0236] ID ARGX-113 buffer pH Additive 1 Additive 2 surfactants F13 150 mg / mL 20 mM HisHCl 6.0 100 mM NaCl 60 mM Sucrose 0.04% w / v PS20 F16 175 mg / mL 20 mM HisHCl 6.0 100 mM NaCl 60 mM Sucrose 0.04% w / v PS20 F17 200 mg / mL 20 mM HisHCl 6.0 100 mM NaCl 60 mM Sucrose 0.04% w / v PS20 F18 175 mg / mL 20 mM HisHCl 6.0 100 mM NaCl 60 mM Sucrose 0.02% w / v PS20 F19 175 mg / mL 20 mM HisHCl 6.0 100 mM NaCl 60 mM Sucrose 0.02% w / v PS80 F20 175 mg / mL 20 mM HisHCl 6.0 100 mM NaCl 60 mM Sucrose 0.05% w / v PS80 F21 175 mg / mL 20 mM HisHCl 6.0 100 mM NaCl 60 mM Sucrose 0.04% w / v PX188

[0237] PS20: Polysorbate 20

[0238] PS80: Polysorbate 80

[0239] PX188: poloxamer 188 (poloxamer 188)

[0240] Experiments were conducted to study 2-week stability at 5 °C, 25 °C, and 40 °C, 1-week shaking stress at 5 °C, 1-week shaking stress at 25 °C, and 5 cycles of freeze / thaw stress for each formulation.

[0241] Buffer exchange and high concentration were performed in 20 mM HisHCl and 100 mM NaCl at a selected pH, followed immediately by the addition of appropriate amounts of sucrose and surfactant stock to achieve the target concentration. To observe any phase transition, the formulations were stored at 5 °C overnight. Phase separation was observed in F17 (200 mg / mL), but it returned to the liquid phase after equilibration at room temperature. The various formulations were then filtered and placed in separate vials.

[0242] After 1 week of storage at 5 °C, and similarly after 5 days of stirring at 5 °C, a reversible solid-liquid phase transition was observed in samples F17, F18, and F19. When warmed to room temperature / equilibrium, the formulations returned to a clear liquid solution. The other formulations (F12, F16, F20, and F21) did not show visible solidification / phase transition under these conditions.

[0243] After 2 weeks of storage at 5 °C, a reversible solid-liquid phase transition was observed in samples F17, F18, and F19. When warmed to room temperature and equilibrated, the formulations returned to a clear liquid solution. The other formulations (F12, F16, F20, and F21) did not show any visible solidification or phase transition under these conditions.

[0244] After 2 weeks of storage at 25 °C, all samples remained in the liquid phase, but many particles were visible in F21.

[0245] After 2 weeks of storage at 40°C, increased opalescence was observed in F13, F18, F19, and F20. Based on the results of Example 2, the increased opalescence observed in F13 was unexpected.

[0246] Fig. 8 This shows that there were no major changes in ARGX-113 protein concentration during this stability study.

[0247] Fig. 9 It shows the osmotic pressure of the formulations after 2 weeks of storage at 5 °C.

[0248] Fig. 10 F13 and F18 show a significant increase in turbidity after 2 weeks of storage at 40 °C.

[0249] Fig. 11A- 11 shows the aggregation results.

[0250] Figs. 12A-12C It shows the iCE results.

[0251] Fig. 13A- 13D It shows subvisible particle results.

[0252] do 8, 9, 10, 11A-11B, 12A-12C, 13A-13D: F1 = F13; F2 = F16; F3 = F17; F4 = F18; F5 = F19; F6 = F20; F7 = F21. do 8, 9, 10, 11A-11B, 12A-12C, 13A-13D The bar shown in order from left to right is as follows (Reference: In Figs. 13A-13DAlthough some bars are very low and few bars are visible on the graph, the order of the bars is the same): F1 initial; F1 shaking at 5°C (shakingsh 5); F1 shaking at 25°C (shakingsh 25); F1 of five cycles of freeze / thaw stress (freeze / thawF / T); F1 at 5°C for 2 weeks (2W5); F1 at 25°C for 2 weeks (2W25); F1 at 40°C for 2 weeks (2W40); F2 initial; F2 shaking at 5°C (shakingsh 5); F2 shaking at 25°C (shakingsh 25); F2 of five cycles of freeze / thaw stress (freeze / thawF / T); F2 at 5°C for 2 weeks (2W5); F2 at 25°C for 2 weeks (2W25); 2 weeks at 40°C (2W40); F3 initial (F3 initial); F3 shaking at 5°C (shakingsh 5); F3 shaking at 25°C (shaking sh 25); F3 of five cycles of freeze / thaw stress (freeze / thawF / T); F3 of 2 weeks at 5°C (2W5); F3 of 2 weeks at 25°C (2W25); 2 weeks at 40°C (2W40); F4 initial (F4 initial); F4 shaking at 5°C (shakingsh 5); F4 of 25°C (shaking sh 25); F4 of five cycles of freeze / thaw stress (freeze / thawF / T); F4 of 2 weeks at 5°C (2W5); F4 of 2 weeks at 25°C (2W25); F4 of 2 weeks at 40°C (2W40); F5 initial; F5 shaking at 5°C; F5 shaking at 25°C; F5 of five cycles of freeze / thaw stress (freeze / thawF / T); F5 for 2 weeks at 5°C (2W5); F5 for 2 weeks at 25°C (2W25); F5 for 2 weeks at 40°C (2W40); F6 initial;F6 shaking at 5°C (shakingsh 6); F6 shaking at 25°C (shaking sh 25); F6 of five cycles of freeze / thaw stress (freeze / thawF / T); F6 for 2 weeks at 5°C (2W5); F6 for 2 weeks at 25°C (2W25); F6 for 2 weeks at 40°C (2W40); F7 initial (F7 initial); F7 shaking at 5°C (shakingsh 5); F7 shaking at 25°C (shaking sh 25); F7 of five cycles of freeze / thaw stress (freeze / thawF / T); F7 for 2 weeks at 5°C (2W5); F7 for 2 weeks at 25°C (2W25); F7 for 2 weeks at 40°C (2W40).;

[0253] From the results obtained in this example, the following conclusions were obtained: (i) Polysorbate 20 was equally effective at 0.02% and 0.04% as to protecting ARGX-113 from shaking and freeze / thaw stress, regardless of protein concentration (e.g., 150 or 175 mg / mL); (ii) Polysorbate 20 and Polysorbate 80 were equally effective as to protecting ARGX-113 from shaking and freeze / thaw stress; (iii) Polysorbate 20 and Poloxamer 188 were equally effective as to protecting ARGX-113 from shaking and freeze / thaw stress; (iv) Aggregation is concentration-dependent, and F16 and F17 (175 mg / mL and 200 mg / mL, respectively) exhibit greater aggregation after 2 weeks of storage at 40 °C compared to F13 (150 mg / mL); (v) ARGX-113 concentrations > 175 mg / mL show a reversible temperature-dependent solid-liquid phase transition; and (vi) lower pH (5.0 and 5.3) show a higher risk of a reversible temperature-dependent solid-liquid phase transition, and pH 5.0 showed a possible risk of chemical degradation forming basic species and possible fragmentation.

[0254] Example 4. Further Excipient Characterization

[0255] This example describes another set of experiments to develop additional ARGX-113 candidate high-concentration formulations and to perform property characterization. In particular, the goal of this set of experiments is to identify candidate high-concentration liquid formulations of ARGX-113 for preclinical toxicology and early phase clinical studies based on specific properties and short-term stability studies.

[0256] The compositions of the seven water-soluble formulations of ARGX-113 studied in this example are shown in Table 5.

[0257] ID ARGX-113 buffer pH Additive 1 Additive 2 Additive 3 surfactants F22 175 mg / mL 20 mM HisHCl 6.0 100 mM ArgCl 60 mM sucrose 10 mM L-methionine 0.03% w / v PS20 F23 175 mg / mL 20 mM HisHCl 6.0 100 mM NaCl 60 mM sucrose 10 mM L-methionine 0.03% w / v PS20 F24 175 mg / mL 20 mM HisHCl 6.0 100 mM ArgCl 60 mM sucrose 10 mM L-methionine 0.03% w / v PS20 F25 175 mg / mL 20 mM HisHCl 6.0 100 mM NaCl 60 mM sucrose 10 mM L-methionine 0.03% w / v PS20 F26 160 mg / mL 20 mM HisHCl 6.0 100 mM ArgCl 60 mM sucrose -- 0.03% w / v PS20 F27 160 mg / mL 20 mM HisHCl 6.0 100 mM NaCl 60 mM sucrose -- 0.03% w / v PS20

[0258] PS20: Polysorbate 20

[0259] F22 and F23 were prepared as 2.0 mL aliquots in vials.

[0260] F24 and F25 were prepared as 2.0 mL aliquots in Nuovo Ompi glass syringes.

[0261] F26 and F27 were prepared as 2.0 mL aliquots in BD SCF Neopak glass syringes.

[0262] For F22, F24, and F26, high concentration was performed in 20 mM HisHCl and 100 mM ArgCl at a selected pH with or without 10 mM L-methionine, followed by large-scale filtration, and for the compounded samples, appropriate amounts of sucrose and surfactant stock solutions were added to achieve the target concentration. The compounded and uncompounded formulations were stored at 5 °C overnight to observe any phase transition. No phase separation was observed in either the compounded or uncompounded formulations. Subsequently, the compounded formulation was filtered, filled, and finished.

[0263] For F23, F25, and F27, high concentration was performed in 20 mM HisHCl and 100 mM NaCl at a selected pH with or without 10 mM L-methionine, followed by bulk filtration, and for the compounded samples, appropriate amounts of sucrose and surfactant stock solutions were added to achieve target concentrations. The compounded and uncompounded formulations were stored at 5 °C overnight to observe any phase transitions. No phase separation was observed in any of the compounded formulations, but phase separation was observed in the uncompounded bulk formulation. The compounded formulations were then filtered, filled, and finished.

[0264] All formulations were then placed in specific storage conditions for a certain period of time before analysis, such as measurements of visible appearance, color, transparency, pH, non-visible particles, purity by SE-HPLC, purity by iCE, purity by CE-SDS, viscosity, and break and glide force.

[0265] All formulations remained in the liquid phase under storage conditions of 3 weeks at 5 °C, 3 weeks at 25 °C, and 3 weeks at 40 °C.

[0266] Although some haze was observed in F22 and F26 after 6 weeks of storage at 40 °C, all formulations remained in the liquid phase under storage conditions of 6 weeks at 5 °C, 6 weeks at 25 °C, and 6 weeks at 40 °C.

[0267] Some precipitation was observed in all formulations stored at 40 °C for 9 weeks, but all formulations remained in the liquid phase under storage conditions of 5 °C for 9 weeks, 25 °C for 9 weeks, and 40 °C for 9 weeks.

[0268] Protein concentrations in each F22-F27 were found to be fundamentally stable under storage conditions including shaking at 5 °C, shaking at 25 °C, freezing / thawing, storage at 5 °C for 3 weeks, storage at 25 °C for 3 weeks, storage at 40 °C for 3 weeks, storage at 5 °C for 6 weeks, storage at 25 °C for 6 weeks, storage at 40 °C for 6 weeks, storage at 5 °C for 9 weeks, storage at 25 °C for 9 weeks, and storage at 40 °C for 9 weeks (within 10% of the initial concentration).

[0269] In each F22-F27, pH was found to be stable under conditions of shaking at 5 °C, shaking at 25 °C, freezing / thawing, storage at 5 °C for 3 weeks, storage at 25 °C for 3 weeks, storage at 40 °C for 3 weeks, storage at 5 °C for 6 weeks, storage at 25 °C for 6 weeks, storage at 40 °C for 6 weeks, storage at 5 °C for 9 weeks, storage at 25 °C for 9 weeks, and storage at 40 °C for 9 weeks.

[0270] Except for 6 weeks of storage at 40°C for F22 and F24, and 9 weeks of storage at 40°C for all formulations, all formulations had virtually no visible particles at all times studied.

[0271] Table 6 shows the osmotic pressure, viscosity, break force, and glide force of various syringe formulations stored at 5 °C for 9 weeks.

[0272] ID Osmolality (mOsmol / kg) Viscosity (cP) Syringeability Break Force Glide Force F24 319 6 4.4 6.4 F25 331 6 4.7 6.6 F26 308 5 4.1 3.8 F27 307 5 4.3 4.2

[0273] Fig. 14 As shown in [figure], turbidity was observed in all formulations stored at 40 °C for 9 weeks, and the NaCl formulation and the formulation without L-methionine showed slightly higher turbidity than the ArgCl formulation.

[0274] Coagulation was evaluated using SE-HPLC. Figs. 15A-15B[Figure] shows the aggregation results. After storage at 40 °C for 9 weeks, a loss of approximately 1.5–1.9% of the main peak was observed depending on the formulation. F23 and F25 (NaCl formulations) showed slightly higher loss of the main peak at 40 °C compared to F22 and F24 (ArgCl formulations). F26 and F27 (without L-methionine) showed slightly higher loss of the main peak at 40 °C compared to formulations with L-methionine. The loss of monomers is mainly due to high molecular weights (MWS) and aggregate formation. F23 and F25 (NaCl formulations) showed slightly higher aggregation at 40 °C compared to F22 and F24 (ArgCl formulations). F26 and F27 (without L-methionine) showed slightly higher HMWs at 40 °C compared to the formulation with L-methionine.

[0275] Figs. 16A-16C This shows the iCE results. After storage at 40 °C for 9 weeks, all formulations showed a similar degree of major peak loss (29–32%). A similar degree of increase in basic and acidic species was observed.

[0276] Figs. 17A-17D It shows subvisible particle results. The data shown from samples stored at 40 °C for 9 weeks are not reliable due to precipitation in these samples.

[0277] From the results obtained from this sample, the following conclusions were obtained: (i) the unprepared ArgCl-containing formulation remained in a liquid state after storage at 5 °C even at a concentration of about 200 mg / mL; (ii) in contrast, the unprepared NaCl-containing formulation did not remain in a liquid state after storage at 5 °C at a concentration of about 200 mg / mL; (iii) the prepared ArgCl-containing formulation did not show precipitation after 6 weeks of storage at 40 °C; (v) the prepared NaCl-containing formulation showed a higher aggregation rate than the ArgCl formulation; and (vi) precipitation of the prepared NaCl-containing formulation was observed after 9 weeks of storage at 40 °C.

[0278] do In 14, 15A-15B, 16A-16C, 17A-17D: F1V = F22; F2V = F23; F1S = F24; F2S = F25; F3S = F26; F4S = F27. In Figs. 14, 15A-15B, 16A-16C, and 17A-17D The bar shown in order from left to right is as follows (Reference: In Figs. 17A-17DAlthough some bars are very low and few are visible on the graph, the order of the bars is the same): F1V initial; F1V shaking at 5°C (shakingsh 5); F1V shaking at 25°C (shakingsh 25); F1V of five cycles of freeze / thaw stress (freeze / thawF / T); F1V at 5°C for 3 weeks (3W5); F1V at 25°C for 3 weeks (3W25); F1V at 40°C for 3 weeks (3W40); F1V at 5°C for 6 weeks (6W5); F1V at 25°C for 6 weeks (6W25); F1V at 40°C for 6 weeks (6W40); F1V at 5°C for 9 weeks (9W5); F1V at 25°C for 9 weeks (9W25); 9 weeks at 40°C (9W40) F1V; F2V initial; shaking F2V at 5°C (shakingsh 5); shaking F2V at 25°C (shakingsh 25); F2V of five cycles of freeze / thaw stress (freeze / thawF / T); 3 weeks at 5°C (3W5) F2V; 3 weeks at 25°C (3W25) F2V; 3 weeks at 40°C (3W40) F2V; 6 weeks at 5°C (6W5) F2V; 6 weeks at 25°C (6W25) F2V; 6 weeks at 40°C (6W40) F2V; 9 weeks at 5°C (9W5) F2V; 9 weeks at 25°C (9W25) F2V; 9 weeks at 40°C (9W40) F2V; F1S initial; shaking F1S at 5°C (shakingsh 5); shaking F1S at 25°C (shakingsh 25); F1S of five cycles of freeze / thaw stress (freeze / thawF / T); 3 weeks at 5°C (3W5) F1S; 3 weeks at 25°C (3W25) F1S; 3 weeks at 40°C (3W40) F1S; 6 weeks at 5°C (6W5) F1S; 6 weeks at 25°C (6W25) F1S; 6 weeks at 40°C (6W40) F1S; 9 weeks at 5°C (9W5) F1S; 9 weeks at 25°C (9W25) F1S;9 weeks at 40°C (9W40) F1S; F2S initial (F2S initial); shaking F2S at 5°C (shakingsh 5); shaking F2S at 25°C (shakingsh 25); F2S of five cycles of freeze / thaw stress (freeze / thawF / T); 3 weeks at 5°C (3W5) F2S; 3 weeks at 25°C (3W25) 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) F2S; 9 weeks at 5°C (9W5) F2S; 9 weeks at 25°C (9W25) F2S; 9 weeks at 40°C (9W40) F2S; F3S initial (F3S initial); shaking F3S at 5°C (shakingsh 5); shaking F3S at 25°C (shakingsh 25); F3S of five cycles of freeze / thaw stress (freeze / thawF / T); 3 weeks at 5°C (3W5) F3S; 3 weeks at 25°C (3W25) F3S; 3 weeks at 40°C (3W40) F3S; 6 weeks at 5°C (6W5) F3S; 6 weeks at 25°C (6W25) F3S; 6 weeks at 40°C (6W40) F3S; 9 weeks at 5°C (9W5) F3S; 9 weeks at 25°C (9W25) F3S; 9 weeks at 40°C (9W40) F3S; F4S initial (F4S initial); shaking F4S at 5°C (shakingsh 5); shaking F4S at 25°C (shakingsh 25); F4S of five cycles of freeze / thaw stress (freeze / thawF / T); 3 weeks at 5°C (3W5) F4S; 3 weeks at 25°C (3W25) F4S; 3 weeks at 40°C (3W40) F4S; 6 weeks at 5°C (6W5) F4S; 6 weeks at 25°C (6W25) F4S; 6 weeks at 40°C (6W40) F4S; 9 weeks at 5°C (9W5) F4S; 9 weeks at 25°C (9W25) F42S; 9 weeks (9W40) F4S at 40°C;

[0279] Example 5. Further Testing for pH Optimization

[0280] This example describes a comparison between two manufacturing methods: Method 1 (pilot) was compared with Method 2 (GMP). Method 2 yields a more accurate result compared to manufacturing 1.

[0281] Several additives were added to WFI (water for injection), dissolved, and then the formulation buffer with other additives was made to a certain volume. The additives were added in a random order.

[0282] The formulation buffer of the result was used during the UF / DF (ultrafiltration / diafiltration) preparation step of the protein (ARGX-113). Polysorbate 20 was not yet added at this time.

[0283] compound Pilot (Method 1) GMP (Method 2) Concentration (g / L) L-Histidine 1.436 1.552 L-Histidine Monohydrochloride 2.252 2.096 Sodium Chloride 5.844 5.84 L-Methionine 1.492 1.492 Sucrose 20.54 20.54

[0284] Next, polysorbate was added through a 10% solution at a 996:4 dilution (e.g., 4008 ml of additive buffer was added to 1000 kg of product formulation buffer), which is called the additive addition step. Therefore, polysorbate 20 (PS20) is added after the diafiltration / ultrafiltration step.

[0285] This results in the following final ARGX-113 formulation:

[0286] 165 mg / mL ARGX-113 in 10 mM L-methionine at pH 6.0 with 20 mM L-histidine / L-histidine hydrochloride, 100 mM sodium chloride, 60 mM sucrose, and 0.04% (w / v) polysorbate 20.

[0287] Those skilled in the art understand that the methods from this embodiment can also be used to make formulations with protein (ARGX-113) concentrations higher than 165 mg / mL, for example, 180 mg / mL, 200 mg / mL, or 300 mg / mL.

[0288] Example 6. Ultra-High Concentration Formulations

[0289] In this example, additional experiments were conducted to evaluate the possibility of viscosity-reducing formulation conditions at higher concentrations of ARGX-113, e.g., 250 to 300 mg / mL. For this purpose, three formulations were prepared at different pH values ​​and ionic strengths.

[0290] Target formulations F101 F102 F103 pH 5.5 6.0 6.5 ARGX-113, mg / mL 250 250 250 Histidine buffer, mM 50 50 50 Arginine, mM 200 200 200 Volume, mL 1 1 1

[0291] Materials and Method

[0292] The ARGX-113 stock solution was buffer-exchanged, followed by high concentration, measurement of protein (ARGX-113) concentration, dilution to approximately 250 mg / mL, and spiking with additives. Each resulting formulation was further divided into 5°C and 25°C storage lots and subsequently analyzed periodically over 14 days for viscosity, osmolality, visual inspection, and filtration testing using a 0.22 μm filter. The analyses were performed on day 0 (D0; day of manufacture), day 3 (D3), day 7 (D7), and day 14 (D4).

[0293] result

[0294] The observed pH, osmotic pressure, protein concentration, viscosity, and visual appearance are shown in Table 9.

[0295] Units F101 F102 F103 pH -- 5.5 6.0 6.5 osmotic pressure mOsm / kg 556 516 469 Protein concentration mg / mL 254 251 272 D0 viscosity measured at 5°C mPA·s 169 95 24 D0 viscosity measured at 25°C mPA·s 110 96 12 D3 viscosity stored at 5°C measured at 5°C mPA·s nd 69 nd D3 viscosity stored at 5°C measured at 25°C mPA·s 4452 2828 1111 Visual appearance on day 7 at 5°C or 25°C - Liquid with gel particles Homogeneous and transparent at both temperatures Homogeneous and transparent at both temperatures Visual appearance on day 14 at 5°C or 25°C - Liquid with gel particles Homogeneous and transparent at both temperatures Homogeneous and transparent at both temperatures D14 filter stored at 5°C - impossibility possible possible D14 viscosity measured at 5°C mPA·s - 17 26 D14 viscosity measured at 25°C mPA·s - 1313 1113

[0296] nd: not done

[0297] In summary:

[0298] dot Three formulations of ARGX-113 with a concentration ≥250 mg / mL were prepared with 200 mM arginine.

[0299] dot The viscosity of the three formulations decreased with pH values. Formulation F103 was the lowest viscosity formulation over 2 weeks at 5 and 25°C (measured at pH=6.5, 24 mPa·s, at 5°C).

[0300] dot The F101 formulation could not be manufactured homogeneously on a small scale due to the formation of gel particles.

[0301] dotThe F102 formulation exhibited unexpected variability in viscosity, which appeared to decrease over time.

[0302] dot The F103 formulation showed low viscosity with fairly reproducible results within two weeks.

[0303] dot The visual appearance of the stored formulations of F102 and F103 remained the same over two weeks whether stored at 5°C or 25°C.

[0304] dot After 7 days, a very viscous, white-like solution was observed in the stock solution containing 100 mM arginine (270-280 mg / mL protein). After 14 days, a solid gel was observed (data not shown).

[0305] dot A filtration test was performed: the viscosity of F102 and F103 remained low (< 26 mPa·s) after filtration at 5°C.

Claims

Claim 1 A water-soluble formulation comprising 90-330 mg / mL of a neonatal Fc receptor (FcRn) antagonist, 20-60 mM histidine / 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, and 0-15 mM L-methionine, wherein the water-soluble formulation has a pH of 6.0-6.5, the FcRn antagonist is composed of a mutant Fc site, the mutant Fc site is composed of two Fc domains forming a homodimer, and the amino acid sequence of each Fc domain is sequence number A water-soluble formulation consisting of the amino acid sequence described in 1 or SEQ ID No.

2. Claim 2 In claim 1, the water-soluble formulation comprises 100-200 mg / mL of an FcRn antagonist, 20 mM histidine / histidine HCl, 50-100 mM NaCl, and 50 mM arginine HCl, and the water-soluble formulation has a pH of 6.

0. Claim 3 In claim 1, the water-soluble formulation comprises 90-220 mg / mL of an FcRn antagonist, 20 mM histidine / histidine HCl, 60 mM sucrose, 90-110 mM NaCl, and 0.02%-0.04% (w / v) polysorbate 20 or polysorbate 80, and has a pH of 6.

0. Claim 4 In claim 1, the water-soluble formulation comprises 100-200 mg / mL of an FcRn antagonist, 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.02%-0.04% (w / v) polysorbate 20 or polysorbate 80, and has a pH of 6.

0. Claim 5 A water-soluble formulation according to claim 1, wherein the water-soluble formulation comprises 150 mg / mL of an FcRn antagonist, 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.04% (w / v) polysorbate 20, has a pH of 6.0, and the amino acid sequence of each Fc domain of the FcRn antagonist consists of the amino acid sequence described in SEQ ID NO.

1. Claim 6 In claim 1, the water-soluble formulation comprises 157.5-192.5 mg / mL of FcRn antagonist, 20 mM histidine / histidine HCl, 60 mM sucrose, 90-110 mM NaCl, and 0.04% (w / v) polysorbate 20, and has a pH of 6.

0. Claim 7 A water-soluble formulation according to claim 1, wherein the water-soluble formulation comprises 175 mg / mL of an FcRn antagonist, 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.04% (w / v) polysorbate 20, has a pH of 6.0, and the amino acid sequence of each Fc domain of the FcRn antagonist consists of the amino acid sequence described in SEQ ID NO.

1. Claim 8 In claim 1, the water-soluble formulation comprises 180-220 mg / mL of FcRn antagonist, 20 mM histidine / histidine HCl, 60 mM sucrose, 90-110 mM NaCl, and 0.04% (w / v) polysorbate 20, and has a pH of 6.

0. Claim 9 A water-soluble formulation according to claim 1, wherein the water-soluble formulation comprises 200 mg / mL of an FcRn antagonist, 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, and 0.04% (w / v) polysorbate 20, has a pH of 6.0, and the amino acid sequence of each Fc domain of the FcRn antagonist consists of the amino acid sequence described in SEQ ID NO.

1. Claim 10 In claim 1, the water-soluble formulation comprises 90-220 mg / mL of an FcRn antagonist, 20 mM histidine / histidine HCl, 60 mM sucrose, 90-110 mM NaCl, 10 mM L-methionine, and 0.02%-0.04% (w / v) of polysorbate 20 or polysorbate 80, and has a pH of 6.

0. Claim 11 In claim 1, the water-soluble formulation comprises 100-200 mg / mL of an FcRn antagonist, 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.02%-0.04% (w / v) of polysorbate 20 or polysorbate 80, and has a pH of 6.

0. Claim 12 In claim 1, the water-soluble formulation comprises 148.5-181.5 mg / mL of FcRn antagonist, 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.04% (w / v) polysorbate 20, and has a pH of 6.

0. Claim 13 In claim 1, the water-soluble formulation comprises 157.5-192.5 mg / mL of FcRn antagonist, 20 mM histidine / histidine HCl, 60 mM sucrose, 90-110 mM NaCl, 10 mM L-methionine, and 0.03% (w / v) polysorbate 20, and has a pH of 6.

0. Claim 14 A water-soluble formulation according to claim 1, wherein the water-soluble formulation comprises 175 mg / mL of an FcRn antagonist, 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.03% (w / v) polysorbate 20, has a pH of 6.0, and the amino acid sequence of each Fc domain of the FcRn antagonist consists of the amino acid sequence described in SEQ ID NO.

1. Claim 15 A water-soluble formulation according to claim 1, wherein the water-soluble formulation comprises 180 mg / mL of an FcRn antagonist, 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.03% (w / v) polysorbate 20, wherein the water-soluble formulation has a pH of 6.0, and the amino acid sequence of each Fc domain of the FcRn antagonist consists of the amino acid sequence described in SEQ ID NO.

1. Claim 16 In claim 1, the water-soluble formulation comprises 180-220 mg / mL of FcRn antagonist, 20 mM histidine / histidine HCl, 60 mM sucrose, 90-110 mM NaCl, 10 mM L-methionine, and 0.03% (w / v) polysorbate 20, and has a pH of 6.

0. Claim 17 A water-soluble formulation according to claim 1, wherein the water-soluble formulation comprises 200 mg / mL of an FcRn antagonist, 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM NaCl, 10 mM L-methionine, and 0.03% (w / v) polysorbate 20, has a pH of 6.0, and the amino acid sequence of each Fc domain of the FcRn antagonist consists of the amino acid sequence described in SEQ ID NO.

1. Claim 18 In claim 1, the water-soluble formulation comprises 100-200 mg / mL of an FcRn antagonist, 50 mM histidine / histidine HCl, 60 mM sucrose, 150 mM arginine HCl, and 0.02%-0.04% (w / v) of polysorbate 20 or polysorbate 80, and has a pH of 6.

0. Claim 19 In claim 1, the water-soluble formulation comprises 100-200 mg / mL of an FcRn antagonist, 20 mM histidine / histidine HCl, 60 mM sucrose, 100 mM arginine HCl, 10 mM L-methionine, and 0.02%-0.04% (w / v) of polysorbate 20 or polysorbate 80, and has a pH of 6.

0. Claim 20 In claim 1, the water-soluble formulation comprises 157.5-192.5 mg / mL of 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, and has a pH of 6.

0. Claim 21 A water-soluble formulation according to claim 1, wherein the water-soluble formulation comprises 175 mg / mL of an 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, has a pH of 6.0, and the amino acid sequence of each Fc domain of the FcRn antagonist consists of the amino acid sequence described in SEQ ID NO.

1. Claim 22 A water-soluble formulation according to claim 1, wherein the water-soluble formulation comprises 180 mg / mL of an 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 water-soluble formulation has a pH of 6.0, and the amino acid sequence of each Fc domain of the FcRn antagonist consists of the amino acid sequence described in SEQ ID NO.

1. Claim 23 In claim 1, the water-soluble formulation comprises 180-220 mg / mL of 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, and has a pH of 6.

0. Claim 24 A water-soluble formulation according to claim 1, wherein the water-soluble formulation comprises 200 mg / mL of an 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, has a pH of 6.0, and the amino acid sequence of each Fc domain of the FcRn antagonist consists of the amino acid sequence described in SEQ ID NO.

1. Claim 25 In claim 1, the water-soluble formulation comprises 100-300 mg / mL of an FcRn antagonist, 50 mM histidine / histidine HCl, and 200 mM arginine HCl, and has a pH of 6.

5. Claim 26 A water-soluble formulation comprising 100-200 mg / mL of an FcRn antagonist, 20 mM L-histidine / L-histidine HCl, 90-110 mM NaCl, 60 mM sucrose, 10 mM L-methionine, and 0.036%-0.044% (w / v) polysorbate 20, wherein the water-soluble formulation has a pH of 6.0-6.5, the FcRn antagonist is composed of a variant Fc site, the variant Fc site is composed of two Fc domains forming a homodimer, and the amino acid sequence of each Fc domain is composed of the amino acid sequence described in SEQ ID NO. 1 or SEQ ID NO.

2. Claim 27 A water-soluble formulation according to claim 26, wherein the amino acid sequence of each Fc domain is composed of the amino acid sequence described in SEQ ID NO.

1. Claim 28 In claim 26, the above-mentioned water-soluble formulation is a water-soluble formulation comprising 162-198 mg / mL of an FcRn antagonist. Claim 29 In claim 26, the water-soluble formulation comprises 180 mg / mL of FcRn antagonist, 20 mM L-histidine / L-histidine HCl, 100 mM NaCl, 60 mM sucrose, 10 mM L-methionine, and 0.04% (w / v) polysorbate 20. Claim 30 A packaged pharmaceutical product comprising a sterile container containing a therapeutically effective amount of a water-soluble formulation of any one of claims 1 to 29. Claim 31 In paragraph 30, the above-mentioned packaged pharmaceutical product is a packaged pharmaceutical product comprising a single-use vial. Claim 32 A device comprising a therapeutically effective amount of a water-soluble formulation of any one of claims 1 to 29. Claim 33 In paragraph 32, the device comprises a syringe containing a water-soluble formulation.

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