Pharmaceutical composition comprising Anti-il-4r antibody

Abstract

Provided are a pharmaceutical formulation, such as a liquid formulation, comprising an anti-IL-4R antibody and a buffer, and use of the pharmaceutical formulation in the treatment of autoimmune diseases. The pharmaceutical formulation is stable, reduces the use of pharmaceutical excipients, and improves the economic benefit and safety of the pharmaceutical product.

Description

Pharmaceutical compositions containing anti-IL-4R antibodies

[0001] Cross-reference to related applications

[0002] This application claims priority and benefit to Patent Application No. 202410691878.8, filed with the State Intellectual Property Office of the People's Republic of China on May 30, 2024, the entire contents of which are hereby incorporated herein by reference: Technical Field

[0003] This disclosure pertains to the field of pharmaceutical formulations, and specifically relates to pharmaceutical compositions comprising anti-IL-4R antibodies and their use in the treatment of autoimmune diseases. Background Technology

[0004] Interleukin-4 (IL-4) is composed of 153 amino acids and has a molecular weight of approximately 17 kDa. It was initially discovered because IL-4 can stimulate B cell proliferation and was named B cell stimulating factor-1 (BSF-1). IL-4 needs to bind to a membrane receptor to exert its biological function. The human interleukin receptor (IL-4R) is a heterodimer formed by two polypeptide chains, one of which, the α chain, has a high affinity for IL-4. Because the IL-4Rα chain plays a dominant role in IL-4 binding in the IL-4R complex, IL-4Rα is often used instead of IL-4R in many scientific studies and reports. IL-4R is expressed on various cell types, including human B cells, mast cells, eosinophils, basophils, macrophages / monocytes, dendritic cells (DCs), fibroblasts, airway epithelium, and smooth muscle cells. IL-4Rα can form two types of receptor complexes with other subunits; in hematopoietic stem cells, the type I receptor, composed of IL-4Rα and γc, is mainly expressed. In non-hematopoietic stem cells, IL-4 primarily functions through the type II receptor composed of IL-4Rα and IL-13Rα1. The type II receptor is a common receptor for both IL-4 and IL-13, with IL-13 binding to IL-13Rα1 to exert its function. Both type I and type II receptors transduce signals through the Jak / STAT pathway. IL-4Rα, γc, and IL-13Rα1 bind to Jak1, Jak3, and Tyk2, respectively, activating downstream pathways. IL-4 and IL-13 can also transduce signals through the insulin receptor substrate family (IRS), ultimately activating nuclear PI3-K and NF-κB. Blocking IL-4R can inhibit the biological functions of both IL-4 and IL-13.

[0005] Like any protein, the biological activity of an antibody depends on the conformational integrity of at least its core amino acid sequence, which protects multiple functional groups of the protein from degradation. Both chemical and physical instabilities can lead to antibody degradation. Because antibodies are larger and more complex than traditional organic and inorganic drugs, their formulation presents unique challenges. Antibody stability can be affected by a variety of factors, such as pH, temperature, repeated freeze / thaw cycles, and shear forces. Active antibodies can lose their activity as a result of physical and chemical instabilities, including denaturation, aggregation (formation of soluble and insoluble aggregates), precipitation, and adsorption, and chemical instabilities, such as racemization, hydrolysis, and deamidation. Any of these instabilities can potentially lead to the formation of antibody byproducts or derivatives with reduced biological activity, increased toxicity, and / or increased immunogenicity.

[0006] In 2017, the U.S. FDA approved Dupixent (dupilumab) for the treatment of moderate to severe eczema (atopic dermatitis) in adult patients. The active ingredient is an antibody that binds to the α subunit of the interleukin-4 receptor (IL-4Rα), a protein that causes inflammation. By binding to this protein, Dupixent inhibits the inflammatory response that plays a role in the development of atopic dermatitis. The drug formulation of dupilumab is as follows: in 2 ml of antibody, it also contains L-arginine hydrochloride (10.5 mg), L-histidine (6.2 mg), polysorbate 80 (4 mg), sodium acetate (2 mg), sucrose (100 mg), and water for injection, pH 5.9.

[0007] Dupilumab uses sucrose, which can promote protein stability in liquid formulations. However, using sucrose alone as an osmotic pressure regulator results in high viscosity formulations with high protein concentrations. Although dupilumab uses arginine hydrochloride in addition to sucrose to reduce viscosity, this makes the formulation more complex, significantly increasing costs and impacting production efficiency.

[0008] Therefore, it is necessary to conduct in-depth research on specific anti-IL-4Rα humanized monoclonal antibodies in order to develop drug formulations suitable for clinical applications.

[0009] Invention Overview

[0010] In a first aspect, this disclosure provides a pharmaceutical composition comprising an anti-IL-4R antibody, a buffer solution, an osmotic regulator, and optionally a surfactant, wherein the anti-IL-4R antibody comprises a heavy chain variable region and a light chain variable region, wherein

[0011] The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO:1;

[0012] The amino acid sequence of the light chain variable region is shown in SEQ ID NO:2;

[0013] Preferably, the anti-IL-4R antibody comprises a heavy chain as shown in SEQ ID NO:3 and a light chain as shown in SEQ ID NO:4.

[0014] In some embodiments, the buffer solution is an aqueous solution of a buffer pair selected from histidine-weak acid buffer pairs, histidine / histidine hydrochloride buffer pairs, acetate / arginine buffer pairs, and acetate / sodium acetate buffer pairs. In some embodiments, the osmotic pressure regulator is a non-saccharide osmotic pressure regulator. In some embodiments, the surfactant is polysorbate 80.

[0015] Secondly, this disclosure provides the use of the pharmaceutical composition described in the first aspect of this disclosure in the preparation of a medicament for treating autoimmune diseases.

[0016] Thirdly, this disclosure provides a method for treating an autoimmune disease in an individual in need, comprising administering to the individual the pharmaceutical composition described in the first aspect of this disclosure.

[0017] Fourthly, this disclosure provides a pharmaceutical composition as described in the first aspect of this disclosure for the treatment of autoimmune diseases.

[0018] Fifthly, this disclosure provides the use of the pharmaceutical compositions described in the first aspect of this disclosure in the treatment of autoimmune diseases.

[0019] Brief description of the attached figures

[0020] Figures 1A to 1D show the relative solubility of GR1802 protein in different buffer solutions in the examples determined by the method of inducing GR1802 protein precipitation with 10% (1A), 20% (1B), 30% (1C), and 40% (1D) PEG3350, respectively.

[0021] Figure 2 shows the relative solubility of GR1802 protein in the examples determined by the method of inducing GR1802 protein precipitation with 30% PEG3350 after adding different osmotic pressure regulators to the H60 formulation of this disclosure.

[0022] Figure 3 shows the viscosity values ​​of the H60 formulation of this disclosure after adding different osmotic pressure regulators.

[0023] Figure 4 shows the effect of osmotic pressure regulators on the viscosity of the formulation.

[0024] Figure 5 shows the effect of the osmotic pressure regulator on the change in the SEC-HPLC polymer ratio after freeze-thaw.

[0025] Figure 6 shows the effect of osmotic pressure regulator on the % change of monomer in SEC-HPLC.

[0026] Figure 7 shows the effect of osmotic pressure regulator on the % change of the basic peak in CEX-HPLC.

[0027] Figure 8 shows the effect of the osmotic pressure regulator on the change in purity % of non-reduced CE-SDS.

[0028] Figure 9 shows the effect of the osmotic pressure regulator on the change in the purity % of reduced CE-SDS.

[0029] Figure 10 shows the peaks of non-reduced CE-SDS in formulations containing different osmotic pressure regulators after being placed at 37°C for 4 weeks.

[0030] Figure 11 shows the peaks of reduced CE-SDS in formulations containing different osmotic pressure regulators after being placed at 37°C for 4 weeks.

[0031] Figure 12 shows the amount of insoluble microparticles (≥2 μm) in H58RE and H58P formulations containing different concentrations of polysorbate 80 during the shaking test.

[0032] Figure 13 shows the amount of insoluble microparticles (≥5 μm) in H58RE and H58P formulations containing different concentrations of polysorbate 80 during the shaking test.

[0033] Figure 14 shows the amount of insoluble microparticles (≥10 μm) in H58RE and H58P formulations containing different concentrations of polysorbate 80 during the shaking test.

[0034] Figure 15 shows the amount of insoluble microparticles (≥25 μm) in H58RE and H58P formulations containing different concentrations of polysorbate 80 during the shaking test.

[0035] Figure 16 shows the SEC-HPLC polymers in H58RE and H58P formulations containing different concentrations of polysorbate 80 during the shaking test.

[0036] Figure 17 shows the SEC-HPLC monomers in formulations with different compositions after being stored at 37°C for 4 weeks.

[0037] Figure 18 shows the SEC-HPLC polymers of formulations with different compositions after being stored at 37°C for 4 weeks.

[0038] Figure 19 shows the percentage of SEC-HPLC monomer peaks (A) and SEC-HPLC polymer peaks (B) in pre-filled syringe formulations with different compositions.

[0039] Information regarding some of the sequences involved in this disclosure is described in the table below:

[0040] Detailed description of the invention

[0041] The principles and features of this disclosure are described below with reference to the accompanying drawings. The examples given are for illustrative purposes only and are not intended to limit the scope of this disclosure.

[0042] definition

[0043] The term “comprising” encompasses both the meaning of “including” and “consisting of”. For example, a composition that “comprising” X may consist of only X or may include other components, such as X+Y.

[0044] The term "optional" means that the event or condition described below may or may not occur, including both the occurrence and non-occurrence of the event or condition. For example, "a pharmaceutical composition contains an optional surfactant" means that the pharmaceutical composition contains a surfactant, and that the pharmaceutical composition does not contain a surfactant.

[0045] The term “about” for the numerical value x means ±10%, unless otherwise specified in the context.

[0046] Each month means approximately every 4 weeks (e.g., every 4 weeks), which is approximately every 28 days (e.g., every 28 days).

[0047] As used herein, the term "antibody" refers to the entire antibody and any antigen-binding fragments or single chains thereof. Naturally occurring "antibodies" are glycoproteins comprising at least two heavy chains (H) and two light chains (L) linked together by disulfide bonds. Each heavy chain contains a heavy chain variable region (VH) and a heavy chain constant region. The heavy chain constant region contains three domains, CH1, CH2, and CH3. Each light chain contains a light chain variable region (VL) and a light chain constant region. The light chain constant region contains one domain, CL. The VH and VL regions can be further subdivided: highly variable regions called hypervariable regions or complementarity-determining regions (CDRs) are interspersed with more conserved regions called framework regions (FRs). Each VH and VL contains three CDRs and four FRs, arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain binding domains that interact with the antigen. The constant region of an antibody can mediate the binding of immunoglobulins to host tissues or factors, including cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system.

[0048] Buffer refers to a buffer solution, buffering agent, or buffer pair. These terms are used interchangeably and all refer to a pair of solutions or reagents that can, to some extent, counteract changes in the pH of a solution caused by a small amount of added strong acid or base, or that do not cause a significant change in the pH of the solution upon slight dilution. It is commonly used, but is not limited to, maintaining a specific pH value of the solution to ensure the stability and efficacy of the contained drug, reducing the impact of pH changes on other components, and helping to control the acidity or alkalinity of the formulation. In pharmaceutical formulations, choosing a suitable buffer solution or buffering agent is crucial because it can reduce damage to the active ingredients in the drug formulation.

[0049] PEG3350 is polyethylene glycol with a molecular weight of 3350.

[0050] As used herein, a “stable” composition refers to a composition in which the protein substantially retains its stability (e.g., physical, chemical, and / or biological activity) under storage conditions. Various analytical techniques for determining protein stability exist in the art and are reviewed in *Peptide and Protein Drug Delivery*, 247-301, edited by Vincent Lee, Marcel Dekker, Inc., New York, NY, Pubs. (1991) and Jones, A. Adv. Drug Delivery Rev. 10:29-90 (1993). Stability can be tested at selected temperatures over selected time periods. A “stable” liquid antibody composition is one in which no significant changes are observed after being kept at refrigerated temperatures (2-8°C) for at least 6 months, 12 months, preferably 2 years, more preferably 3 years; or at room temperature (23-27°C) for at least 3 months, preferably 6 months, more preferably 1 year; or under stress conditions (~40°C) for at least 1 month, preferably 3 months, more preferably 6 months. Various stability standards can be used, such as a degradation rate of no more than 10%, preferably 5%, of the antibody monomer (e.g., determined by SEC purity, RP-HPLC purity, CEX purity, CE-SDS purity (non-reducing), etc.). Alternatively, stability can be indicated if the solution remains clear to slightly milky white by visual analysis or by using a suspension meter.

[0051] The term "CEX purity" as used in this article refers to the percentage of the dominant peak in the CEX and can be used to assess the stability of pharmaceutical compositions containing the GR1802 protein. The charge heterogeneity of GR1802 is evaluated by determining the percentage of acidic and basic variants using CEX.

[0052] The term "SEC purity" as used herein refers to the percentage of monomers in SEC and can be used to assess the stability of pharmaceutical compositions containing the GR1802 protein. SEC was used to separate the monomer GR1802 from aggregates and fragments based on size under non-denaturing conditions. The sum of peaks eluted before the main peak was reported as the percentage of aggregates, and the sum of peaks eluted after the main peak was reported as the percentage of degradation products.

[0053] The term "CE-SDS purity" as used herein refers to the percentage of intact antibody in CE-SDS, which can be used to assess the stability of pharmaceutical compositions containing GR1802 protein. CE-SDS was used to separate byproducts and degradation products from intact GR1802 based on molecular size under non-reducing conditions. The sum of peaks separated from the main peak was reported as the percentage of impurities.

[0054] The detection methods for SEC-HPLC, CEX-HPLC, NrCE-SDS, and rCE-SDS in this disclosure are as follows:

[0055] Purity determination by SEC-HPLC (size exclusion chromatography) was performed using a TSKgel G3000SWXL column and a high-performance liquid chromatograph. The mobile phase was 50 mM PB / 300 mM NaCl solution at pH 7.0. After sample dilution, the sample was injected and isocratic eluted at a flow rate of 1.0 ml / min for 15 min. The sample was detected at 280 nm UV wavelength, and the peak area percentage of monomers and polymers was obtained by peak area normalization.

[0056] CEX-HPLC (cation exchange chromatography) purity determination was performed using MabPac TM SCX-10BioLC TM An Analytical column and a high-performance liquid chromatograph were used. Mobile phase A consisted of 10 mM MES + 10 mM HEPES at pH 5.0; mobile phase B consisted of 10 mM MES + 10 mM HEPES at pH 10.0; mobile phase C consisted of 1.0 M sodium chloride solution; and mobile phase D consisted of water. Samples were diluted and injected, and nonlinear gradient elution was performed at a flow rate of 0.8 mL / min for 40 min. Detection was performed at 280 nm UV wavelength, and the peak area percentages of acidic peaks, the main peak, and the basic peaks were obtained using peak area normalization.

[0057] Both non-reducing CE-SDS (sodium dodecyl sulfate capillary electrophoresis) (NrCE-SDS) and reduced CE-SDS (rCE-SDS) methods were performed using a capillary electrophoresis apparatus. An uncoated capillary column with an effective length of 20 cm and an SDS-MW gel separation gel were used. The column temperature was 25℃, the separation voltage was 15 kV, and the detection wavelength was 214 nm. The purity of the samples was determined under both non-reducing and reducing conditions.

[0058] Differential scanning fluorescence (DSF) was measured using a Real-Time PCR instrument. The sample was diluted, mixed with a fluorescent dye (Orange Protein Gel Stain, Life), and transferred to a PCR tube. A temperature program was set (25°C to 95°C, 1°C / min). Fluorescence was collected and data recorded in real time. The derivative of the fluorescence-temperature curve was calculated; the highest peak of the derivative curve is the temperature at time T. m value.

[0059] The term “viscosity” as used herein refers to the physical property of the pharmaceutical composition itself, which is measured using a microvisometer, but other methods may also be used for measurement.

[0060] As used herein, the term “freeze-thaw stability” refers to the ability of a pharmaceutical composition to maintain its physical, chemical, and / or biological properties during freezing and thawing cycles.

[0061] The term “pKa” used in this article refers to the acidity coefficient or the dissociation constant of a drug. In chemistry and biochemistry, it refers to a specific equilibrium constant that represents the ability of an acid to dissociate hydrogen ions.

[0062] Anti-IL-4R antibody (GR1802 antibody / GR1802 protein)

[0063] The anti-IL-4R antibody disclosed herein is also known as GR1802 antibody and GR1802 protein. Chinese Patent Application No. 201810360234.5 has conducted preliminary research on anti-IL-4R antibodies, describing their structure, amino acid sequence, preparation method, and biological activity. This disclosure is a follow-up study of that Chinese patent application, the entire contents of which are hereby incorporated herein by reference.

[0064] In a first aspect, the antigen-binding fragment for binding human IL-4R described in this disclosure comprises:

[0065] Heavy chain variable region and light chain variable region, wherein:

[0066] The amino acid sequence of the variable region of the heavy chain of the antibody is shown in SEQ ID NO:1;

[0067] The amino acid sequence of the variable region of the antibody light chain is shown in SEQ ID NO:2.

[0068] In some embodiments, the amino acid sequence of the heavy chain variable region of the antibody is shown in SEQ ID NO:1, and the amino acid sequence of the light chain variable region of the antibody is shown in SEQ ID NO:2.

[0069] In some embodiments, the anti-IL-4R antibody comprises a heavy chain as shown in SEQ ID NO:3 and a light chain as shown in SEQ ID NO:4.

[0070] In some embodiments of the first aspect, the antibody is a whole antibody, a Fab fragment, an F(ab')2 fragment, or a single-chain Fv fragment (scFv).

[0071] In some embodiments of the first aspect, the antibody is a fully human antibody.

[0072] In some embodiments of the first aspect, the antibody further comprises a heavy chain constant region selected from IgG1, IgG2 or IgG4 subtypes and / or a light chain constant region selected from κ or λ subtypes.

[0073] Pharmaceutical Composition

[0074] This disclosure provides pharmaceutical compositions comprising the anti-IL-4R antibody described herein.

[0075] In a first aspect, this disclosure provides a pharmaceutical composition comprising the anti-IL-4R antibody and a buffer solution. In some embodiments, the anti-IL-4R antibody is the GR1802 protein.

[0076] In some embodiments, the pharmaceutical composition further comprises an osmotic pressure regulator. In some embodiments, the pharmaceutical composition further comprises an osmotic pressure regulator and a surfactant.

[0077] In some embodiments, the buffer solution is an aqueous solution of a buffer pair formed by histidine and a weak acid. In some embodiments, the buffer solution is an aqueous solution of a buffer pair formed by histidine and a weak acid with a lower acidity than histidine. In some embodiments, the buffer pair formed by histidine and a weak acid, or the buffer pair formed by histidine and a weak acid with a lower acidity than histidine, is selected from histidine / acetic acid buffer pair, histidine / glutamate buffer pair, or histidine / aspartic acid buffer pair; preferably, the buffer pair formed by histidine and a weak acid, or the buffer pair formed by histidine and a weak acid with a lower acidity than histidine, is a histidine / glutamate buffer pair.

[0078] In some embodiments, the buffer solution is an aqueous solution selected from buffer pairs such as histidine / histidine hydrochloride buffer pair, acetate / arginine buffer pair, and acetate / sodium acetate buffer pair.

[0079] In some embodiments, the concentration of the buffer solution is from about 10 mM to about 50 mM, for example, about 10 mM, about 15 mM, about 20 mM, about 25 mM, about 30 mM, about 35 mM, about 40 mM, about 45 mM and about 50 mM.

[0080] In some embodiments, the osmotic pressure regulator is a non-carbohydrate osmotic pressure regulator. In some embodiments, the non-carbohydrate osmotic pressure regulator is selected from proline, glutamic acid, arginine and acetic acid, arginine and glutamic acid, or histidine and aspartic acid. Preferably, the non-carbohydrate osmotic pressure regulator is proline, arginine and acetic acid, or arginine and glutamic acid; more preferably, the non-carbohydrate osmotic pressure regulator is arginine and glutamic acid.

[0081] In some embodiments, the surfactant is polysorbate 80, or the surfactant is at least about 0.01%, for example, about 0.01% to about 0.10% of polysorbate 80, or the surfactant is at least about 0.02%, for example, about 0.02% to about 0.10% of polysorbate 80, where % refers to weight / volume percentage or %w / v.

[0082] In some embodiments, the pH of the pharmaceutical composition is about 4.5 to about 6.5 or about 5.0 to about 6.5; for example, pH is about 5.0, about 5.1, about 5.2, about 5.3, about 5.4, about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, about 6.0, about 6.1 or about 6.2; preferably, pH is about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, about 6.0, about 6.1 or about 6.2; more preferably, pH is about 5.8.

[0083] In some embodiments, the pharmaceutical composition comprises about 20 mg / ml to about 190 mg / ml of an anti-IL-4R antibody, a buffer pair formed by histidine and a weak acid weaker than histidine, a non-carbohydrate osmoregulator, and water. In some embodiments, the pharmaceutical composition comprises about 20 mg / ml to about 190 mg / ml of an anti-IL-4R antibody, a buffer pair formed by histidine and a weak acid weaker than histidine, a non-carbohydrate osmoregulator, and water, wherein the buffer pair formed by histidine and a weak acid weaker than histidine is selected from histidine / acetic acid buffer pairs, histidine / glutamate buffer pairs, or histidine / aspartic acid buffer pairs, and the non-carbohydrate osmoregulator is selected from proline, arginine and acetic acid, arginine and glutamate, or histidine and aspartic acid.

[0084] In some embodiments, the pharmaceutical composition comprises about 20 mg / ml to about 190 mg / ml of an anti-IL-4R antibody, a buffer pair formed by histidine and a weak acid weaker than histidine, arginine, glutamic acid, and water. In some embodiments, the buffer pair is a histidine / glutamate buffer pair, and the osmolarity regulator is arginine and glutamate; therefore, the pharmaceutical composition may comprise about 20 mg / ml to about 190 mg / ml of an anti-IL-4R antibody, histidine, arginine, glutamate, and water. In some embodiments, the buffer pair is a histidine / acetic acid buffer pair, and the osmolarity regulator is arginine and acetic acid; therefore, the pharmaceutical composition may comprise about 20 mg / ml to about 190 mg / ml of an anti-IL-4R antibody, histidine, arginine, acetic acid, and water.

[0085] In some embodiments, the pharmaceutical composition comprises about 20 mg / ml to about 500 mg / ml of anti-IL-4R antibody, histidine, arginine, glutamic acid, polysorbate 80, and water.

[0086] In some embodiments, the pharmaceutical composition comprises about 20 mg / ml to about 190 mg / ml of anti-IL-4R antibody, about 10 mM to about 50 mM of histidine, about 100 mM to about 300 mM of arginine, about 100 mM to about 300 mM of glutamic acid, about 0.1 mg / ml to about 1.0 mg / ml of polysorbate 80, and water.

[0087] In some embodiments, the concentration of histidine is from about 10 mM to about 50 mM, for example about 10 mM, about 15 mM, about 20 mM, about 25 mM, about 30 mM, about 35 mM, about 40 mM, about 45 mM and about 50 mM.

[0088] In some embodiments, the concentration of arginine is from about 100 mM to about 300 mM, for example, about 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 174, 175, 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, 270, 275, 280, 285, 290, 295 or 300 mM.

[0089] In some embodiments, the concentration of arginine is from about 100 mM to about 300 mM, for example, about 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 156, 160, 165, 170, 174, 175, 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, 270, 275, 280, 285, 290, 295 or 300 mM.

[0090] In some embodiments, the concentration of the polysorbate 80 is at least about 0.01%, for example about 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, or 0.10%.

[0091] In some embodiments, the pharmaceutical composition comprises about 150 mg / ml of anti-IL-4R antibody, about 20 mM histidine, about 174 mM arginine, about 156 mM glutamic acid, at least about 0.2 mg / ml of polysorbate 80, and water.

[0092] In some embodiments, the pharmaceutical composition comprises an anti-IL-4R antibody at a concentration of about 20 mg / ml to about 500 mg / ml, about 20 mg / ml to about 190 mg / ml, or about 150 mg / ml, histidine / histidine hydrochloride, proline, and water.

[0093] In some embodiments, the pharmaceutical composition comprises about 150 mg / ml of anti-IL-4R antibody, 20 mM histidine / histidine hydrochloride, 25 mg / ml proline, 0.02% polysorbate 80, and water, and the pH of the pharmaceutical composition is 5.5-6.2 or 5.5-5.8 or 5.8.

[0094] In some embodiments, the pharmaceutical composition comprises an anti-IL-4R antibody at a concentration of about 20 mg / ml to about 500 mg / ml, about 20 mg / ml to about 190 mg / ml, or about 150 mg / ml, acetic acid, arginine, glutamic acid, and water.

[0095] In some embodiments, the pharmaceutical composition comprises about 150 mg / ml of anti-IL-4R antibody, 20 mM acetic acid, arginine, 25 mg / ml glutamic acid, 0.02% polysorbate 80 and water, and the pH of the pharmaceutical composition is 5.2-5.5 or 5.2.

[0096] In some embodiments, the pharmaceutical composition comprises an anti-IL-4R antibody at a concentration of about 20 mg / ml to about 500 mg / ml, about 20 mg / ml to about 190 mg / ml, or about 150 mg / ml, acetate / sodium acetate, proline, and water.

[0097] In some embodiments, the pharmaceutical composition comprises about 150 mg / ml of anti-IL-4R antibody, 20 mM acetate / sodium acetate, 25 mg / ml proline, 0.02% polysorbate 80, and water, and the pH of the pharmaceutical composition is 5.0-5.5 or 5.2.

[0098] In some embodiments, the pharmaceutical composition may also contain the remainder water.

[0099] In some embodiments, the pharmaceutical composition is free of chloride ions and / or sucrose.

[0100] In some embodiments, the pharmaceutical compositions of this disclosure have any one of the following characteristics:

[0101] a. After storage at 37±2°C for 8 weeks, the SEC-HPLC monomer percentage remains at least about 94.7%, and / or, after storage at 37±2°C for 8 weeks, the SEC-HPLC polymer percentage remains at most about 2.8%;

[0102] b. After storage at 37±2°C for 6 weeks, the SEC-HPLC monomer percentage remains at least about 95.4%, and / or, after storage at 37±2°C for 6 weeks, the SEC-HPLC polymer percentage remains at most about 2.7%;

[0103] c. After storage at 37±2°C for 4 weeks, the SEC-HPLC monomer percentage remains at least approximately 96.4%, and / or, after storage at 37±2°C for 4 weeks, the SEC-HPLC polymer percentage remains at most approximately 2.5%; or

[0104] d. After storage at 37±2°C for 2 weeks, the SEC-HPLC monomer percentage remains at least about 97.3%, and / or, after storage at 37±2°C for 2 weeks, the SEC-HPLC polymer percentage remains at most about 2.2%.

[0105] Secondly, this disclosure provides the use of the pharmaceutical composition described in the first aspect of this disclosure in the preparation of a medicament for treating autoimmune diseases.

[0106] Thirdly, this disclosure provides a method for treating an autoimmune disease in an individual in need, comprising administering to the individual the pharmaceutical composition described in the first aspect of this disclosure.

[0107] Fourthly, this disclosure provides a pharmaceutical composition as described in the first aspect of this disclosure for the treatment of autoimmune diseases.

[0108] Fifthly, this disclosure provides the use of the pharmaceutical compositions described in the first aspect of this disclosure in the treatment of autoimmune diseases.

[0109] The disclosed pharmaceutical composition containing a specific anti-IL-4Rα humanized monoclonal antibody is sucrose-free and maintains stability. In particular, acetic acid, glutamic acid, and aspartic acid can form buffer pairs with histidine and can also act as osmotic regulators with arginine and / or histidine, which reduces the use of pharmaceutical excipients compared to Dupixent formulations and significantly improves the cost-effectiveness and safety of the pharmaceutical product. Example

[0110] The partial sequence information of the IL-4R antibody (i.e., GR1802 protein) in the examples is as follows:

[0111] The IL-4R antibody contains both heavy chain variable regions and light chain variable regions.

[0112] The heavy chain variable region sequence of the IL-4R antibody is SEQ ID NO:1, and the light chain variable region sequence is SEQ ID NO:2.

[0113] The heavy chain sequence of the IL-4R antibody is as follows:

[0114] The light chain sequence of the IL-4R antibody is as follows:

[0115] Example 1: Effect of buffer salt type and pH of formulation on the solubility of drug formulations containing anti-IL-4R antibodies.

[0116] Table 1 shows the pKa and buffer range of commonly used acetate buffers, histidine buffers, citrate buffers, and phosphate buffers in the pharmaceutical field.

[0117] Table 1

[0118] Without adding other excipients, a drug formulation of GR1802 protein was prepared by mixing a certain amount of GR1802 protein with acetate buffer, histidine buffer and citrate buffer at a concentration of 20 mM and a pH range of 4.5 to 6.5, in order to investigate the effect of buffer salt type and pH of the formulation on the solubility of GR1802 protein.

[0119] 1.1 Composition and Numbering of the Formulation

[0120] A formulation of IL-4R monoclonal antibody (i.e. GR1802 protein) with different types of buffer salts and formulation pH was prepared (batch number: GR1802-20180917). The composition, pH and number of the formulation are shown in Table 2. The concentration of GR1802 protein is approximately 17.3 mg / ml.

[0121] Table 2

[0122] / indicates that the formulation does not contain the corresponding excipient.

[0123] 1.2 Experimental Methods

[0124] ① 10% PEG3350 (polyethylene glycol) induces precipitation

[0125] 50 μl of formulation sample, 5 μl of 10× formulation buffer, 20 μl of 50% PEG3350 and 25 μl of water were mixed to prepare an induction precipitation solution containing 10% PEG3350. The solution was incubated overnight at 4°C to induce GR1802 protein precipitation. The supernatant was collected by centrifugation and mixed with water at a ratio of 1:1. The protein concentration in the supernatant of each formulation sample was determined.

[0126] ② 20% PEG3350 (polyethylene glycol) induced precipitation

[0127] 50 μl of sample, 5 μl of 10× formulation buffer, 40 μl of 50% PEG3350 and 5 μl of water were mixed to prepare an induction precipitation solution containing 20% ​​PEG3350. The solution was incubated overnight at 4°C to induce GR1802 protein precipitation. The supernatant was collected by centrifugation and mixed with water at a ratio of 1:1. The protein concentration in the supernatant of each formulation sample was determined.

[0128] ③ 30% PEG3350 (polyethylene glycol) induced precipitation

[0129] To prepare an induction precipitation solution containing 30% PEG3350, 30 μl of sample, 7 μl of 10× formulation buffer, 60 μl of 50% PEG3350, and 3 μl of water were mixed. The solution was incubated overnight at 4°C to induce GR1802 protein precipitation. The supernatant was collected by centrifugation and mixed with water at a 1:1 ratio. The protein concentration in the supernatant of each formulation sample was then determined.

[0130] ④40% PEG3350 (polyethylene glycol) induced precipitation

[0131] To prepare an induction precipitation solution containing 40% PEG3350, 10 μl of sample, 9 μl of 10× formulation buffer, 80 μl of 50% PEG3350, and 1 μl of water were mixed. The solution was incubated overnight at 4°C to induce GR1802 protein precipitation. The supernatant was collected by centrifugation and mixed with water at a 1:1 ratio. The protein concentration in the supernatant of each formulation sample was then determined.

[0132] 1.3 Experimental Results

[0133] The relative solubility of GR1802 protein in different formulations was determined by precipitation induction with 10%, 20%, 30%, and 40% PEG3350, respectively. The experimental results are shown in Figures 1A to 1D and Tables 3 to 6.

[0134] Table 3. Concentration (mg / ml) of GR1802 protein in the supernatant after 10% PEG3350-induced precipitation.

[0135] Table 4. Concentration (mg / ml) of GR1802 protein in the supernatant after precipitation induced by 20% PEG3350

[0136] Table 5. Concentration (mg / ml) of GR1802 protein in the supernatant after precipitation induced by 30% PEG3350

[0137] Table 6. Concentration (mg / ml) of GR1802 protein in the supernatant after 40% PEG3350-induced precipitation.

[0138] - indicates that the formulation does not contain the corresponding excipient.

[0139] After inducing precipitation with 10% PEG3350, the concentration of GR1802 protein in the supernatant of the formulations using acetate buffer and histidine buffer at different pH values ​​was close to the theoretical concentration (8.64 mg / ml) for which precipitation did not occur. This indicates that most of the GR1802 protein did not precipitate in either of these two buffers. However, the concentration of GR1802 protein in the supernatant of the formulation using citrate buffer decreased to below 2 mg / ml, indicating that most of the GR1802 protein in the sample of the formulation using citrate buffer precipitated after induction with 10% PEG3350.

[0140] Increasing the concentration of PEG3350 to induce precipitation to 20% resulted in near-complete precipitation of GR1802 protein in formulations using acetate buffer at pH 6.0 and 6.5, while the formulation using histidine buffer showed virtually no precipitation.

[0141] When the concentration of PEG3350 used to induce precipitation was further increased to 30%, some GR1802 protein precipitated in the formulation using acetate buffer at pH 5.5, while almost no precipitation occurred in the formulation using histidine buffer.

[0142] When the concentration of PEG3350 used to induce precipitation was finally increased to 40%, a significant amount of GR1802 protein precipitated in formulations using acetate buffer at pH 5.0–6.5, while in formulations using histidine buffer, only a portion of GR1802 protein precipitated in the pH 5.5 formulation.

[0143] Based on the above results, the GR1802 protein has the best solubility in the histidine buffer, followed by the acetate buffer, and the worst solubility in the citrate buffer.

[0144] Example 2: Effects of different osmotic pressure regulators on the solubility of GR1802 protein and the viscosity and stability of formulations containing histidine salt buffer.

[0145] 2.1 Preliminary investigation of the solubility of GR1802 protein and the viscosity of formulations containing histidine salt buffers using different osmotic regulators.

[0146] Different osmotic pressure regulators (25 mg / ml proline, 27 mg / ml arginine hydrochloride, 17 mg / ml glycine, and 75 mg / ml trehalose) were added to formulation H60 (20 mM histidine / histidine hydrochloride, pH 6.0), along with a certain amount of GR1802 protein, to prepare formulation samples with a GR1802 protein concentration of approximately 150 mg / ml. The relative solubility of GR1802 protein in each formulation was determined using a 30% PEG3350-induced precipitation method, and the viscosity of each formulation sample at 20°C was also measured. The results are shown in Figures 2 and 3. The results show that proline, glycine, and trehalose can all increase the solubility of GR1802 protein, but arginine hydrochloride significantly reduces the solubility of GR1802 protein. Glycine and trehalose have virtually no effect on reducing the viscosity of the obtained formulations, proline can reduce the viscosity of the formulation by about one-third, while arginine hydrochloride can reduce the viscosity of the formulation by about half.

[0147] In summary, adding proline to the formulation can simultaneously improve the solubility of GR1802 protein and reduce the viscosity of the formulation, while arginine hydrochloride, although it can significantly reduce the viscosity of the formulation, also reduces the solubility of GR1802 protein.

[0148] 2.2. Investigation of the effects of different osmotic pressure regulators on the viscosity and stability of formulations containing histidine salt buffers.

[0149] 2.2.1, Scheme

[0150] Based on the experimental results in Example 1, the GR1802 protein exhibits high solubility in histidine buffer. Therefore, this experiment primarily investigated formulations containing histidine buffer. Different osmotic pressure regulators (trehalose, sucrose, proline, arginine / trehalose hydrochloride, arginine / acetic acid, and arginine / glutamic acid) and 0.2 mg / ml polysorbate 80 were added to 20 mM histidine buffer. The pH of each formulation was 5.8, and the concentration of GR1802 protein was approximately 150 mg / ml. The viscosity and Tg of each formulation were measured. m The viscosity and stability of formulations containing GR1802 protein were compared by performing freeze-thaw stability tests and high-temperature (37°C) stability tests, and by using different osmotic pressure regulators.

[0151] 2.2.2 Components:

[0152] The composition of the formulation (batch number: GR1802-20181123) is shown in Table 7.

[0153] Table 7. Composition of the formulation √: indicates that the formulation contains the corresponding excipient; -: indicates that the formulation does not contain the corresponding excipient.

[0154] 2.2.3 Experimental Methods:

[0155] Freeze-thaw test: Samples of each formulation were placed in the center of a 100-well square box, with 2ml EP tubes (filled with water) placed in the remaining wells. Two square boxes filled with 2ml EP tubes (filled with water) were stacked vertically. A freeze-thaw cycle consisted of freezing at -80℃ for 24 hours followed by thawing at room temperature for 24 hours. Three freeze-thaw cycles were performed. This method simulates a slow-freezing and slow-thawing process. The polymer ratio of each sample before and after freeze-thaw was determined using SEC-HPLC.

[0156] High-temperature (37°C) stability test: Samples of each formulation were placed under high-temperature (37°C) conditions for stability testing to accelerate the degradation of GR1802 protein, thereby allowing for the assessment of the stability of GR1802 protein in different formulations within a short period of time. The testing conditions and indicators for the high-temperature stability test are shown in Table 8.

[0157] Table 8. Examination conditions and indicators for high temperature stability test

[0158] 2.2.4 Results and Analysis:

[0159] Viscosity

[0160] The viscosity of each formulation sample at 20°C was measured using a microvisometer to investigate the effect of the type of osmotic pressure regulator on the viscosity of the formulation. The results are shown in Table 9 and Figure 4.

[0161] Table 9. Effect of osmotic pressure regulators on formulation viscosity - indicates that the formulation does not contain the corresponding excipient.

[0162] The results showed that in 20mM histidine / acetic acid buffer and 20mM histidine / glutamate buffer at pH 5.8, arginine / acetic acid and arginine / glutamate significantly reduced the viscosity of the formulation, proline slightly reduced the viscosity, while sugars (trehalose, sucrose) increased the viscosity. According to literature reports, to eject 1 ml of injection solution from a pre-filled syringe with a 27G needle within 10 seconds under a thrust of 30 N, the viscosity of the formulation needs to be controlled below approximately 12 mPa·s. Therefore, proline, arginine / acetic acid, and arginine / glutamate are suitable as osmotic pressure regulators.

[0163] freeze-thaw stability

[0164] A freeze-thaw cycle was defined as room temperature → -80℃ → room temperature, with three slow freeze-thaw cycles. The polymer ratio of each formulation sample before and after freeze-thaw was determined using SEC-HPLC. The effect of the type of osmotic pressure regulator on the increase in polymer ratio after freeze-thaw was investigated. The results are shown in Table 10 and Figure 5.

[0165] Table 10. Effect of osmotic pressure regulators on the change in the proportion of SEC-HPLC polymers after freeze-thaw cycles. - indicates that the formulation does not contain the corresponding excipient.

[0166] The results show that the addition of various osmotic pressure regulators to 20mM histidine / histidine hydrochloride buffer, 20mM histidine / acetic acid buffer, and 20mM histidine / glutamate buffer at pH 5.8 can inhibit the aggregation of GR1802 protein during freeze-thaw cycles to a certain extent, with arginine / glutamate showing the best inhibitory effect.

[0167] High temperature (37℃) stability

[0168] The stability of formulations using trehalose, sucrose, proline, arginine hydrochloride + trehalose, arginine / acetic acid, and arginine / glutamate as osmotic pressure regulators in 20mM histidine / histidine hydrochloride buffer (pH 5.8), 20mM histidine / acetic acid buffer (pH 5.8), and 20mM histidine / glutamate buffer (pH 5.8) at high temperature (37°C) was compared. The results are shown in Tables 11, 12, 13, and 14, and Figures 6 to 11.

[0169] Table 11. Effect of osmotic pressure regulators on high-temperature stability: SEC-HPLC monomer % - indicates that the formulation does not contain the corresponding excipient.

[0170] Table 12. Effect of osmotic pressure regulator on high-temperature stability: CEX-HPLC alkaline peak % - indicates that the formulation does not contain the corresponding excipient.

[0171] Table 13. Effect of osmotic pressure regulators on high-temperature stability: Purity % of non-reduced CE-SDS - indicates that the formulation does not contain the corresponding excipient.

[0172] Table 14. Effect of osmotic pressure regulator on high-temperature stability: Purity of reduced CE-SDS (%) - indicates that the formulation does not contain the corresponding excipient.

[0173] The results showed that the GR1802 protein was prone to hydrolysis and fragmentation in formulations using trehalose, sucrose, and trehalose / arginine hydrochloride as osmolarity regulators. After 4 weeks of storage at 37°C, these three formulations exhibited a high proportion of fragment peaks following the monomer peak in SEC-HPLC, abnormal acidic peaks in the CEX-HPLC chromatograms, and a high proportion of specific fragment peaks in both the non-reduced CE-SDS chromatogram (Figure 10) and the reduced CE-SDS chromatogram (Figure 11). Among the other formulations, the formulations using arginine / glutamate as osmolarity regulators showed a slower rate of increase in SEC-HPLC polymers and a slower rate of decrease in CE-SDS purity. This indicates that using sugars as osmolarity regulators in a 20mM histidine / histidine hydrochloride buffer system at pH 5.8 leads to abnormal fragmentation and degradation of the GR1802 protein under high-temperature conditions. The formulations using arginine / glutamate as osmolarity regulators showed the best stability under high-temperature conditions, followed by proline, and then arginine / acetic acid.

[0174] Based on the above results, the Tm values ​​of all formulations are above 55℃, indicating that the GR1802 protein can maintain its undenatured native conformation under high temperature conditions of 37℃. At the same time, under this condition, the interaction between GR1802 protein molecules is enhanced and the chemical degradation rate is accelerated. Therefore, the stability results of each formulation under this condition can reflect its stability under long-term storage conditions (2-8℃) to a certain extent. The use of arginine / glutamic acid, proline, and arginine / acetic acid can all maintain the GR1802 protein in the formulation in a good stable state.

[0175] In a 20 mM histidine / histidine hydrochloride (pH 5.8) buffer, formulations using sugars such as trehalose and sucrose as osmotic regulators exhibited relatively high viscosity and were prone to hydrolysis and fragmentation at high temperatures (37°C). Combinations of arginine with two weak acids (i.e., arginine / acetic acid and arginine / glutamate) significantly reduced the viscosity of the formulation, lowering the viscosity of a GR1802 protein concentration of 150 mg / ml to approximately 6 mPa·s. In particular, the arginine / glutamate combination showed the best effect in inhibiting GR1802 protein aggregation during freeze-thaw cycles and reducing purity at high temperatures (37°C).

[0176] In summary, in the GR1802 formulation, arginine / glutamic acid and arginine / acetic acid, as osmotic pressure regulators, can not only significantly reduce the viscosity of the formulation, but also inhibit the aggregation of GR1802 protein during freeze-thaw cycles and improve the stability of the formulation under high temperature (37°C) conditions. Proline, as an osmotic pressure regulator, can also reduce the viscosity of the formulation, inhibit the aggregation of GR1802 protein during freeze-thaw cycles, and improve the stability of the formulation under high temperature (37°C) conditions.

[0177] Example 3: Effect of surfactant concentration in formulation on the content of insoluble microparticles and the proportion of SEC-HPLC polymers in the formulation.

[0178] 3.1 Experimental Methods

[0179] To improve the stability of the formulation, polysorbate 80 was selected as the surfactant, and the effect of polysorbate 80 concentration on the content of insoluble microparticles and the proportion of SEC-HPLC polymers in the formulation was investigated. The concentration of polysorbate 80 and the composition of the formulation are shown in Table 15.

[0180] Table 15. Concentration and composition of polysorbate 80

[0181] In Table 15, "%" refers to weight / volume percentage or %w / v. For example, 0.05% polysorbate 80 means that the content of polysorbate 80 in the formulation is 0.5 mg / mL.

[0182] The experimental protocol for investigating the effect of polysorbate 80 concentration on formulation stability is shown in Table 16.

[0183] Table 16. Experimental protocols for the effect of polysorbate 80 concentration on formulation stability.

[0184] 3.2 Experimental Results

[0185] The results of the shaking experiment are shown in Figures 12-16. The results show that the GR1802 formulation without polysorbate 80, after shaking at room temperature for 4 days, had significantly higher numbers of insoluble particles and a higher proportion of SEC-HPLC polymers, indicating that the GR1802 formulation is easily affected by mechanical forces such as shaking. When 0.005%–0.10% concentrations of polysorbate 80 were added to the GR1802 formulation, and after shaking for 4 days at room temperature, when the concentration of polysorbate 80 in formulation H58RE increased to ≥0.01%, the number of insoluble particles and the proportion of SEC-HPLC polymers in the formulation were significantly lower than those in the formulation without polysorbate 80. After shaking tests, formulation H58P had a higher number of insoluble particles and a higher proportion of polymers in SEC-HPLC than formulation H58RE, indicating that formulation H58P was less protective against mechanical forces on GR1802 protein than formulation H58RE. However, when the concentration of polysorbate 80 was ≥0.02%, the difference between the two formulations became smaller, or even almost negligible.

[0186] Therefore, ≥0.01% polysorbate 80 is added to the GR1802 formulation H58RE to inhibit the aggregation of GR1802 protein caused by mechanical forces. Considering that polysorbate 80 may undergo adsorption and degradation during production and storage, ≥0.02% polysorbate 80 is preferably added to the GR1802 formulation, i.e., the polysorbate 80 content is ≥0.2 mg / ml. This disclosure provides a stable GR1802 formulation, effectively improving product quality.

[0187] Example 4: Further study on the composition of the formulation

[0188] 4.1 Composition of the formulation

[0189] The composition of GR1802 formulation (batch number: GR1802-20190131) is shown in Table 17.

[0190] Table 17. Composition of GR1802 formulation √: indicates that the formulation contains the corresponding excipient; -: indicates that the formulation does not contain the corresponding excipient.

[0191] In Table 17, "%" refers to weight / volume percentage or %w / v. For example, 0.02% polysorbate 80 means that the content of polysorbate 80 in the formulation is 0.2 mg / mL.

[0192] 4.2. Examination Conditions and Indicators

[0193] The evaluation conditions and indicators for the formulation are shown in Table 18.

[0194] Table 18. Evaluation conditions and indicators for GR1802 formulation

[0195] 4.3 Results

[0196] The stability of the formulations with the best stability among the four formulation systems (histidine / arginine / glutamic acid, histidine / histidine hydrochloride / proline, acetic acid / arginine / glutamic acid, and acetic acid / sodium acetate / proline) under high temperature (37℃) conditions was compared. The results are shown in Tables 19 and 20 (Figures 17 and 18).

[0197] Table 19. Formulation Comparison: SEC-HPLC Monomer %

[0198] Table 20. Formulation Comparison: SEC-HPLC Polymer %

[0199] The results showed that the ratio of SEC-HPLC monomers and SEC-HPLC polymers increased at the slowest rate in the histidine / arginine / glutamic acid / pH5.8 formulation.

[0200] The high-temperature stability of the four formulation systems in buffers at different pH values ​​was further investigated, and the results are shown in Table 21 below.

[0201] Table 21. High-temperature stability test results of GR1802 formulation

[0202] Histidine / arginine / glutamic acid preparations with a pH range of 5.5–6.2, acetic acid / sodium acetate / proline preparations with a pH range of 5.0–5.5, and histidine / hydrochloric acid histidine / proline preparations with a pH range of 5.5–5.8 can all reduce the viscosity of GR1802 preparations with a protein concentration of 150 mg / ml to about 6–7 mPa·s. The histidine / arginine / glutamic acid preparation with a pH of 5.8 showed the slowest increase in the proportion of SEC-HPLC polymers under high temperature (37°C) conditions.

[0203] Example 5: Stability Study of Pre-filled Syringe Formulation

[0204] The clinical indication for GR1802 is inflammatory diseases associated with type 2 hypersensitivity reactions, requiring long-term administration. The intended clinical route of administration is subcutaneous injection, and pre-filled syringe formulations are typically developed for ease of administration. The silicone oil coated inside the pre-filled glass syringe cannula to reduce friction may cause aggregation of the GR1802 protein in the formulation, and other exudates may also cause degradation of the GR1802 protein. Therefore, it is necessary to investigate the stability of GR1802 formulations, such as formulation H58RE (20mM histidine buffer + arginine + glutamate + 0.2mg / ml polysorbate 80, pH 5.8), in pre-filled syringe formulations. A high-temperature (37°C) stability test was designed to investigate the stability of GR1802 formulations, such as formulations H58RE and H58P.

[0205] 5.1 Composition of the formulation

[0206] The composition of the formulation used in the stability study of the pre-filled syringe is shown in Table 22 (batch number: GR1802-20190627). The concentration of histidine in the H58RE formulation is 20 mM, i.e., 3.113 mg / ml. When the concentration of glutamate is 23.0 mg / ml and the concentration of arginine is 24.0 mg / ml, the osmotic pressure of the GR1802 formulation is within the isotonic range of 285-310 mOsmol / kg, and the pH of the formulation is approximately 5.8.

[0207] Table 22. Composition of the formulation

[0208] In Table 22, “%” means weight / volume percentage or %w / v. For example, 0.02% polysorbate 80 means that the content of polysorbate 80 in the formulation is 0.2 mg / mL.

[0209] 5.2. Examination Conditions and Indicators

[0210] The conditions and indicators for the stability study of the pre-filled syringe formulation are shown in Table 23.

[0211] Table 23. Stability Study Conditions and Indicators for Pre-filled Syringe Formulations

[0212] 5.3 Results

[0213] As shown in Figure 19, after 8 weeks of storage at high temperature (37℃), the proportion of SEC-HPLC polymers in formulation H58RE increased by 0.7%, while the proportion of SEC-HPLC monomer peaks decreased by 4%. In formulation H58P, the rate of increase in the proportion of SEC-HPLC polymers was slightly faster, while the rate of decrease in SEC-HPLC monomer purity was slightly faster; the proportion of SEC-HPLC polymers increased by 1.4%, while the proportion of SEC-HPLC monomer peaks decreased by 4.5%.

[0214] Through the investigation of various amino acids and sugars, arginine / glutamic acid, arginine / acetic acid, or proline can be selected as osmotic pressure regulators and stabilizers for GR1802 formulations. These can effectively reduce the viscosity of GR1802 formulations and inhibit the aggregation of GR1802 protein in the formulation. The concentration of histidine buffer was determined to be 20 mM. At this concentration, the formulation buffer can maintain the pH of the formulation and can be rapidly neutralized to the physiological pH when it enters the human body for clinical use.

[0215] In the description of this specification, the terms "one embodiment," "some embodiments," "specific embodiment," etc., refer to a specific feature, structure, material, or characteristic described in connection with that embodiment or example, which is included in at least one embodiment or example of this disclosure. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples.

[0216] The above description is only a preferred embodiment of this disclosure and is not intended to limit this disclosure. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this disclosure should be included within the protection scope of this disclosure.

Claims

1. A pharmaceutical composition comprising an anti-IL-4R antibody, a buffer, an osmotic pressure adjusting agent, and optionally a surfactant, wherein the anti-IL-4R antibody comprises a heavy chain variable region and a light chain variable region, wherein the amino acid sequence of the heavy chain variable region is set forth in SEQ ID NO: 1; the amino acid sequence of the light chain variable region is set forth in SEQ ID NO: 2; preferably, the anti-IL-4R antibody comprises a heavy chain as set forth in SEQ ID NO: 3 and a light chain as set forth in SEQ ID NO:

4.

2. The pharmaceutical composition of claim 1, further comprising a surfactant.

3. The pharmaceutical composition of any one of claims 1-2, wherein the buffer is an aqueous solution of a buffer pair formed by histidine and a weak acid, or, the buffer is an aqueous solution of a buffer pair formed by histidine and a weak acid that is weaker than histidine.

4. The pharmaceutical composition of claim 3, wherein the buffer pair formed by histidine and a weak acid or the buffer pair formed by histidine and a weak acid that is weaker than histidine is selected from the group consisting of a histidine / acetic acid buffer pair, a histidine / glutamic acid buffer pair, or a histidine / aspartic acid buffer pair; preferably, the buffer pair formed by histidine and a weak acid or the buffer pair formed by histidine and a weak acid that is weaker than histidine is a histidine / glutamic acid buffer pair.

5. The pharmaceutical composition of any one of claims 1-2, wherein the buffer is an aqueous solution of a buffer pair selected from the group consisting of a histidine / histidine hydrochloride buffer pair, an acetic acid / arginine buffer pair, and an acetic acid / sodium acetate buffer pair.

6. The pharmaceutical composition of any one of claims 1-5, wherein the osmotic pressure adjusting agent is a non-sugar osmotic pressure adjusting agent.

7. The pharmaceutical composition of claim 6, wherein the non-sugar osmotic pressure adjusting agent is selected from the group consisting of proline, glutamic acid, arginine and acetic acid, arginine and glutamic acid, or histidine and aspartic acid; preferably, the non-sugar osmotic pressure adjusting agent is proline, arginine and acetic acid, or arginine and glutamic acid, more preferably, the non-sugar osmotic pressure adjusting agent is arginine and glutamic acid.

8. The pharmaceutical composition of any one of claims 2-7, wherein the surfactant is polysorbate 80, or, the surfactant is at least about 0.01%, e.g., about 0.01%-about 0.10% of polysorbate 80, or, the surfactant is at least about 0.02%, e.g., about 0.02%-about 0.10% of polysorbate 80, wherein % means weight / volume percent or % w / v.

9. The pharmaceutical composition of any one of claims 1-8, wherein the pH of the pharmaceutical composition is about 4.5-about 6.5 or about 5.0-about 6.5; e.g., the pH is about 5.0, about 5.1, about 5.2, about 5.3, about 5.4, about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, about 6.0, about 6.1, or about 6.

2. ​ ​ ​ 10. The pharmaceutical composition of any one of claims 1, 2, 8-9, comprising about 20 mg / ml to about 190 mg / ml of the anti-IL-4R antibody, histidine, arginine, glutamic acid, and water.

11. The pharmaceutical composition of any one of claims 1 and 9, comprising about 20 mg / ml to about 190 mg / ml of the anti-IL-4R antibody, about 10 mM to about 50 mM of histidine, about 100 mM to about 300 mM of arginine, about 100 mM to about 300 mM of glutamic acid, about 0.1 mg / ml to about 1.0 mg / ml of polysorbate 80, and water.

12. The pharmaceutical composition of any one of claims 1 and 9, comprising about 150 mg / ml of the anti-IL-4R antibody, about 20 mM histidine, about 174 mM arginine, about 156 mM glutamic acid, at least about 0.2 mg / ml of polysorbate 80, e.g., about 0.2 mg / ml to about 1.0 mg / ml of polysorbate 80, and water.

13. The pharmaceutical composition of any one of claims 1, 2, 8-9, comprising about 20 mg / ml to about 500 mg / ml, about 20 mg / ml to about 190 mg / ml, or about 150 mg / ml of the anti-IL-4R antibody, histidine / histidine hydrochloride, proline, and water.

14. The pharmaceutical composition of any one of claims 1, 2, 8-9, comprising about 20 mg / ml to about 500 mg / ml, about 20 mg / ml to about 190 mg / ml, or about 150 mg / ml of the anti-IL-4R antibody, acetic acid, arginine, glutamic acid, and water.

15. The pharmaceutical composition of any one of claims 1, 2, 8-9, comprising about 20 mg / ml to about 500 mg / ml, about 20 mg / ml to about 190 mg / ml, or about 150 mg / ml of the anti-IL-4R antibody, acetic acid / sodium acetate, proline, and water.

16. The pharmaceutical composition of any one of claims 1-15, which is free of chloride ions and / or sucrose.

17. The pharmaceutical composition of any one of claims 1, 8-9, and 16, comprising about 20 mg / ml to about 500 mg / ml of the anti-IL-4R antibody, histidine, arginine, glutamic acid, polysorbate 80, and water.

18. The pharmaceutical composition of any one of claims 1-17, having any one of the following properties: a. SEC-HPLC monomer % is at least about 94.7% after storage at 37 ± 2 °C for 8 weeks, and / or SEC-HPLC % of aggregates is at most about 2.8% after storage at 37 ± 2 °C for 8 weeks; b. SEC-HPLC monomer % is at least about 95.4% after storage at 37 ± 2 °C for 6 weeks, and / or SEC-HPLC % of aggregates is at most about 2.7% after storage at 37 ± 2 °C for 6 weeks; c. After storage at 37±2°C for 4 weeks, the SEC-HPLC monomer percentage remains at least approximately 96.4%, and / or, after storage at 37±2°C for 4 weeks, the SEC-HPLC polymer percentage remains at most approximately 2.5%; or d. After storage at 37±2°C for 2 weeks, the SEC-HPLC monomer percentage remains at least about 97.3%, and / or, after storage at 37±2°C for 2 weeks, the SEC-HPLC polymer percentage remains at most about 2.2%.

19. Use of the pharmaceutical composition of any one of claims 1 to 18 in the preparation of a medicament for treating autoimmune diseases.

20. A method of treating an individual with an autoimmune disease, comprising administering to said individual the pharmaceutical composition of any one of claims 1 to 18.

21. A pharmaceutical composition according to any one of claims 1 to 18 for the treatment of autoimmune diseases.

22. Use of the pharmaceutical composition according to any one of claims 1 to 18 in the treatment of autoimmune diseases.

Images