Anti-IL-4R alpha antibody preparation as well as preparation method and application thereof

Abstract

The invention provides an anti-IL-4R alpha antibody preparation and a preparation method and application thereof, the antibody preparation comprises 10-200 mg / mL anti-IL-4R alpha antibody, a buffer agent, a stabilizer and a surfactant, and the pH value of the antibody preparation is 5.0-7.0. The antibody preparation provided by the invention has better stability under the conditions of high temperature, room temperature, illumination and the like, can be kept in a liquid form and is convenient to use.

Description

Technical Field

[0001] The present invention belongs to the field of biological preparations, and specifically relates to an anti-IL-4Rα antibody preparation and a preparation method and application thereof. Background Art

[0002] Interleukin-4 (IL-4, also known as B cell stimulating factor or BSF-1) was originally characterized by its ability to stimulate B cell proliferation in response to low concentrations of anti-surface immunoglobulin antibodies. IL-4 has been shown to have a wide range of biological activities, including growth stimulation of T cells, mast cells, granulocytes, megakaryocytes, and erythrocytes. IL-4 induces the expression of class II major histocompatibility complex molecules in resting B cells and enhances the secretion of IgE and IgG1 isotypes by stimulated B cells. The biological activities of IL-4 are mediated by specific cell surface IL-4 receptors.

[0003] Anti-IL-4R antibodies can be used to treat or prevent diseases such as atopic dermatitis, asthma and other conditions. In order to realize the use of anti-IL-4R antibodies in treating diseases, in-depth research on anti-IL-4R antibodies is needed to develop stable formulations suitable for clinical applications. Summary of the invention

[0004] The present invention provides a stable antibody preparation comprising an antibody that can specifically bind to human interleukin-4 receptor α (hIL-4Rα).

[0005] In some embodiments, the antibody formulation of the present invention comprises 10-200 mg / mL of an anti-IL-4Rα antibody, as well as a buffer, a stabilizer, and a surfactant; the anti-IL-4Rα antibody comprises a heavy chain variable region as shown in SEQ ID NO: 1 and a light chain variable region as shown in SEQ ID NO: 2. In some embodiments, the heavy chain of the anti-IL-4Rα antibody comprises a sequence as shown in SEQ ID NO: 3, and the light chain of the anti-IL-4Rα antibody comprises a sequence as shown in SEQ ID NO: 4. In some embodiments, the anti-IL-4Rα antibody is dupilumab.

[0006] In some embodiments, the pH of the antibody formulation is 5.0-7.0.

[0007] In some embodiments, the stabilizer is selected from one or more of sucrose, trehalose, mannitol, arginine or a pharmaceutically acceptable salt thereof, glycine or a pharmaceutically acceptable salt thereof, proline or a pharmaceutically acceptable salt thereof, methionine or a pharmaceutically acceptable salt thereof. In some embodiments, the stabilizer comprises glycine, proline, or a combination of proline and methionine or a pharmaceutically acceptable salt thereof. In some embodiments, the stabilizer comprises proline or a pharmaceutically acceptable salt thereof. In some embodiments, the stabilizer comprises proline or a pharmaceutically acceptable salt thereof and methionine or a pharmaceutically acceptable salt thereof.

[0008] In some embodiments, the concentration of the stabilizer is 50-500 mM. In some embodiments, the concentration of the stabilizer is 100-450 mM. In some embodiments, the concentration of the stabilizer is 120-350 mM. In some embodiments, the concentration of the stabilizer is 145-330 mM. In some embodiments, the concentration of the stabilizer is about 50 mM, about 60 mM, about 70 mM, about 80 mM, about 90 mM, about 100 mM, about 110 mM, about 120 mM, about 130 mM, about 146 mM, about 150 mM, about 154 mM, about 160 mM, about 164 mM, about 170 mM, about 180 mM, about 190 mM, about 201 mM, about 217 mM, about 220 mM, about 227 mM, about about 230mM, about 234mM, about 237mM, about 240mM, about 247mM, about 250mM, about 260mM, about 270mM, about 280mM, about 290mM, about 293mM, about 300mM, about 320mM, about 325mM, about 350mM, about 380mM, about 400mM, about 450mM, about 500mM, or a range (including endpoints) between any two of these values ​​or any value therein.

[0009] In some embodiments, the stabilizer comprises 100-300 mM proline or a pharmaceutically acceptable salt thereof. In some embodiments, the stabilizer comprises 180-230 mM proline or a pharmaceutically acceptable salt thereof. In some embodiments, the stabilizer comprises about 217 mM proline or a pharmaceutically acceptable salt thereof. In some embodiments, the stabilizer comprises about 191 mM proline or a pharmaceutically acceptable salt thereof.

[0010] In some embodiments, the stabilizer comprises 100-300mM proline or a pharmaceutically acceptable salt thereof and 1-30mM methionine or a pharmaceutically acceptable salt thereof. In some embodiments, the stabilizer comprises 180-230mM proline or a pharmaceutically acceptable salt thereof and 5-15mM methionine or a pharmaceutically acceptable salt thereof. In some embodiments, the stabilizer comprises about 217mM proline or a pharmaceutically acceptable salt thereof and about 10mM methionine or a pharmaceutically acceptable salt thereof. In some embodiments, the stabilizer comprises about 191mM proline or a pharmaceutically acceptable salt thereof and about 10mM methionine or a pharmaceutically acceptable salt thereof.

[0011] In some embodiments, the stabilizer comprises 20-100mM sucrose and 101-300mM arginine or a pharmaceutically acceptable salt thereof. In some embodiments, the stabilizer comprises 35-90mM sucrose and 101-245mM arginine or a pharmaceutically acceptable salt thereof. In some embodiments, the pharmaceutically acceptable salt of arginine is arginine hydrochloride or arginine acetate. In some embodiments, the stabilizer comprises about 44mM sucrose and about 110mM arginine hydrochloride. In some embodiments, the stabilizer comprises about 88mM sucrose and about 237mM arginine hydrochloride. In some embodiments, the stabilizer further comprises 1-30mM methionine. In some embodiments, the concentration of methionine is about 10mM. In some embodiments, the stabilizer comprises about 44mM sucrose, about 110mM arginine hydrochloride and about 10mM methionine. In some embodiments, the stabilizer further comprises 0.01-0.5mM EDTA. In some embodiments, the concentration of EDTA is about 0.05 mM.

[0012] In some embodiments, the pH of the antibody formulation is 5.3-6.5. In some embodiments, the pH of the antibody formulation is 5.9 ± 0.3. In some embodiments, the pH of the antibody formulation is about 5.0, about 5.3, about 5.6, about 5.7, about 5.8, about 5.9, about 6.0, about 6.1, about 6.2, about 6.3, about 6.4, about 6.5, about 6.8, about 7.0, or a range (including endpoints) between any two of these values ​​or any value therein.

[0013] In some embodiments, the buffer is selected from histidine, citric acid, histidine-acetic acid and histidine-citric acid. In some embodiments, the concentration of the buffer is 1-80mM. In some embodiments, the concentration of the buffer is 5-80mM. In some embodiments, the concentration of the buffer is 10-80mM. In some embodiments, the concentration of the buffer is 10-60mM. In some embodiments, the concentration of the buffer is 15-60mM. In some embodiments, the concentration of the buffer is 20-55mM. In some embodiments, the concentration of the buffer is about 1mM, about 3mM, about 5mM, about 8mM, about 10mM, about 12mM, about 15mM, about 16mM, about 18mM, about 19mM, about 20mM, about 21mM, about 22mM, about 23mM, about 25mM, about 27mM, about 30mM, about 35mM, about 40mM, about 50mM, about 60mM, about 70mM, about 80mM, or the range (including endpoints) between any two values ​​in these numerical values ​​or any value therein. In some embodiments, the buffer is about 20mM histidine buffer. In some embodiments, the histidine buffer includes histidine and histidine hydrochloride. In some embodiments, the buffer comprises about 20mM histidine and 26±15mM acetic acid. In some embodiments, the buffer comprises about 20 mM histidine and 26±5 mM acetate.

[0014] In some embodiments, the concentration of the anti-IL-4Rα antibody is 20-200 mg / mL. In some embodiments, the concentration of the anti-IL-4Rα antibody is 40-200 mg / mL. In some embodiments, the concentration of the anti-IL-4Rα antibody is 20-150 mg / mL. In some embodiments, the concentration of the anti-IL-4Rα antibody is 100-200 mg / mL. In some embodiments, the concentration of the anti-IL-4Rα antibody is 120-180 mg / mL. In some embodiments, the concentration of the anti-IL-4Rα antibody is 145-180 mg / mL. In some embodiments, the concentration of the anti-IL-4Rα antibody is 20-80 mg / mL. In some embodiments, the concentration of the anti-IL-4Rα antibody in the antibody preparation is about 20 mg / mL, about 30 mg / mL, about 40 mg / mL, about 47 mg / mL, about 50 mg / mL, about 60 mg / mL, about 70 mg / mL, about 80 mg / mL, about 90 mg / mL, about 100 mg / mL, about 110 mg / mL, about 120 mg / mL, about 130 mg / mL, about 140 mg / mL, about 150 mg / mL, about 160 mg / mL, about 170 mg / mL, about 175 mg / mL, about 180 mg / mL, about 190 mg / mL, about 200 mg / mL, or a range (including endpoints) between any two of these values ​​or any value therein. In some embodiments, the concentration of the anti-IL-4Rα antibody in the antibody preparation is about 150 mg / mL. In some embodiments, the concentration of the anti-IL-4Rα antibody in the antibody formulation is about 175 mg / mL.

[0015] In some embodiments, the surfactant is polysorbate or poloxamer. In some embodiments, the surfactant is polysorbate 80, polysorbate 20 or poloxamer 188. In some embodiments, the surfactant is polysorbate 80. In some embodiments, the concentration of the surfactant is 0.1-5mg / mL. In some embodiments, the concentration of the surfactant is 0.5-5mg / mL. In some embodiments, the concentration of the surfactant is 0.5-3mg / mL. In some embodiments, the concentration of the surfactant is about 0.1mg / mL, 0.2mg / mL, 0.3mg / mL, 0.5mg / mL, 1mg / mL, 2mg / mL, 3mg / mL, 5mg / mL, or the range (including endpoints) between any two values ​​in these numerical values ​​or any value therein. In some embodiments, the surfactant is 0.5-3mg / mL polysorbate 80. In some embodiments, the surfactant is 1-2mg / mL polysorbate 80. In some embodiments, the surfactant is about 2 mg / mL polysorbate 80.

[0016] In some embodiments, the antibody formulation is free or substantially free of acetate.

[0017] In some embodiments, the antibody formulation does not contain arginine or a pharmaceutically acceptable salt thereof.

[0018] In some embodiments, the antibody formulation comprises 20-200 mg / mL anti-IL-4Rα antibody, 5-80 mM buffer, 200-450 mM stabilizer and surfactant. In some embodiments, the antibody formulation comprises 20-200 mg / mL anti-IL-4Rα antibody, 5-80 mM buffer, 210-450 mM stabilizer and surfactant.

[0019] In some embodiments, the present invention provides an antibody formulation comprising an anti-IL-4Rα antibody, wherein the antibody formulation comprises 20-200 mg / mL anti-IL-4Rα antibody, 5-80 mM buffer, 100-450 mM stabilizer, 0.1-5 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody formulation is 5.3-6.5.

[0020] In some embodiments, the antibody formulation comprises 20-100 mg / mL anti-IL-4Rα antibody, 10-30 mM buffer, 120-350 mM stabilizer, 0.5-3 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody formulation is 5.3-6.5.

[0021] In some embodiments, the antibody formulation comprises 45-55 mg / mL anti-IL-4Rα antibody, 10-30 mM buffer, 145-330 mM stabilizer, 1-3 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody formulation is 5.3-6.5.

[0022] In some embodiments, the antibody formulation comprises 100-200 mg / mL anti-IL-4Rα antibody, 5-80 mM buffer, 120-350 mM stabilizer, 0.5-3 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody formulation is 5.3-6.5.

[0023] In some embodiments, the antibody formulation comprises 130-200 mg / mL anti-IL-4Rα antibody, 10-60 mM buffer, 145-330 mM stabilizer, 0.5-3 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody formulation is 5.3-6.5.

[0024] In some embodiments, the buffer is selected from histidine, citric acid, histidine-acetic acid and histidine-citric acid. In some embodiments, the stabilizer is selected from glycine or a pharmaceutically acceptable salt thereof, proline or a pharmaceutically acceptable salt thereof, a combination of proline or a pharmaceutically acceptable salt thereof and methionine or a pharmaceutically acceptable salt thereof. In some embodiments, the stabilizer is selected from a combination of glycine, proline, proline and methionine. In some embodiments, the anti-IL-4Rα antibody comprises a heavy chain variable region shown in SEQ ID NO: 1 and a light chain variable region shown in SEQ ID NO: 2.

[0025] In some embodiments, the antibody formulation comprises 20-200 mg / mL anti-IL-4Rα antibody, a buffer, proline, polysorbate, or poloxamer.

[0026] In some embodiments, the antibody formulation comprises 20-200 mg / mL anti-IL-4Rα antibody, 5-80 mM buffer, proline, 0.1-5 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody formulation is 5.3-6.5.

[0027] In some embodiments, the antibody formulation comprises 20-200 mg / mL anti-IL-4Rα antibody, 5-80 mM buffer, 100-300 mM proline, 0.1-5 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody formulation is 5.3-6.5.

[0028] In some embodiments, the antibody formulation comprises 20-200 mg / mL anti-IL-4Rα antibody, 5-80 mM buffer, 100-300 mM proline, 0.5-3 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody formulation is 5.3-6.5.

[0029] In some embodiments, the antibody formulation comprises 45-55 mg / mL anti-IL-4Rα antibody, 10-30 mM buffer, 150-250 mM proline, 1-3 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody formulation is 5.3-6.5.

[0030] In some embodiments, the antibody formulation comprises 100-200 mg / mL anti-IL-4Rα antibody, 5-80 mM buffer, 150-250 mM proline, 0.5-3 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody formulation is 5.3-6.5.

[0031] In some embodiments, the antibody formulation comprises 20-200 mg / mL anti-IL-4Rα antibody, a buffer, proline, methionine, polysorbate, or poloxamer.

[0032] In some embodiments, the antibody formulation comprises 20-200 mg / mL anti-IL-4Rα antibody, 5-80 mM buffer, proline, methionine, 0.1-5 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody formulation is 5.3-6.5.

[0033] In some embodiments, the antibody formulation comprises 20-200 mg / mL anti-IL-4Rα antibody, 5-80 mM buffer, 100-300 mM proline, 5-30 mM methionine, 0.1-5 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody formulation is 5.3-6.5.

[0034] In some embodiments, the antibody formulation comprises 20-200 mg / mL anti-IL-4Rα antibody, 5-80 mM buffer, 100-300 mM proline, 5-30 mM methionine, 0.5-3 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody formulation is 5.3-6.5.

[0035] In some embodiments, the antibody formulation comprises 45-55 mg / mL anti-IL-4Rα antibody, 10-30 mM buffer, 150-250 mM proline, 5-30 mM methionine, 1-3 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody formulation is 5.3-6.5.

[0036] In some embodiments, the antibody formulation comprises 100-200 mg / mL anti-IL-4Rα antibody, 5-80 mM buffer, 150-250 mM proline, 5-30 mM methionine, 0.5-3 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody formulation is 5.3-6.5.

[0037] In some embodiments, the antibody formulation comprises 20-200 mg / mL anti-IL-4Rα antibody, 5-80 mM buffer, 20-100 mM sucrose, 101-300 mM arginine or a pharmaceutically acceptable salt thereof, 0.1-5 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody formulation is 5.3-6.5.

[0038] In some embodiments, the antibody preparation comprises 20-200 mg / mL anti-IL-4Rα antibody, 5-80 mM buffer, 20-100 mM sucrose, 101-300 mM arginine hydrochloride, 0.5-3 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody preparation is 5.3-6.5.

[0039] In some embodiments, the antibody formulation comprises 45-55 mg / mL anti-IL-4Rα antibody, 10-30 mM buffer, 35-90 mM sucrose, 101-245 mM arginine hydrochloride, 1-3 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody formulation is 5.3-6.5.

[0040] In some embodiments, the antibody preparation comprises 100-200 mg / mL anti-IL-4Rα antibody, 5-80 mM buffer, 35-90 mM sucrose, 101-245 mM arginine hydrochloride, 0.5-3 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody preparation is 5.3-6.5.

[0041] In some embodiments, the buffer is selected from the group consisting of histidine, citric acid, histidine-acetic acid, and histidine-citrate.In some embodiments, the anti-IL-4Rα antibody comprises a heavy chain variable region as shown in SEQ ID NO:1 and a light chain variable region as shown in SEQ ID NO:2.

[0042] In some embodiments, the antibody preparation comprises about 50 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, about 146 mM sucrose (equivalent to about 5% sucrose), about 2 mg / mL polysorbate 80 (equivalent to about 0.2% polysorbate 80), and the pH of the antibody preparation is 5.3-6.5.

[0043] In some embodiments, the antibody formulation comprises about 50 mg / mL anti-IL-4Rα antibody, about 20 mM citric acid, about 146 mM sucrose, about 2 mg / mL polysorbate 80, and the pH of the antibody formulation is 5.3-6.5.

[0044] In some embodiments, the antibody formulation comprises about 50 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, about 234 mM sucrose (equivalent to about 8% sucrose), about 2 mg / mL polysorbate 80, and the pH of the antibody formulation is 5.9±0.3.

[0045] In some embodiments, the antibody formulation comprises about 50 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, about 225 mM trehalose (equivalent to about 8.5% trehalose dihydrate), about 2 mg / mL polysorbate 80, and the pH of the antibody formulation is 5.9±0.3.

[0046] In some embodiments, the antibody formulation comprises about 50 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, about 293 mM glycine (equivalent to about 2.2% glycine), about 2 mg / mL polysorbate 80, and the pH of the antibody formulation is 5.9±0.3.

[0047] In some embodiments, the antibody formulation comprises about 50 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, about 247 mM mannitol (equivalent to about 4.5% mannitol), about 2 mg / mL polysorbate 80, and the pH of the antibody formulation is 5.9±0.3.

[0048] In some embodiments, the antibody preparation comprises about 50 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, about 237 mM arginine hydrochloride (equivalent to about 5% arginine hydrochloride), and about 2 mg / mL polysorbate 80, and the pH of the antibody preparation is 5.9±0.3.

[0049] In some embodiments, the antibody preparation comprises about 50 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, about 88 mM sucrose (equivalent to about 3% sucrose), about 237 mM arginine hydrochloride, and about 2 mg / mL polysorbate 80, and the pH of the antibody preparation is 5.9±0.3.

[0050] In some embodiments, the antibody formulation comprises about 50 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, about 217 mM proline (equivalent to about 2.5% proline), about 2 mg / mL polysorbate 80, and the pH of the antibody formulation is 5.9±0.3.

[0051] In some embodiments, the antibody preparation comprises about 175 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, about 110 mM arginine hydrochloride, about 44 mM sucrose (equivalent to about 1.5% sucrose), and about 1 mg / mL polysorbate 80 (equivalent to about 0.1% polysorbate 80), and the pH of the antibody preparation is 5.9±0.3.

[0052] In some embodiments, the antibody formulation comprises about 175 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, about 110 mM arginine hydrochloride, about 44 mM sucrose, about 0.05 mM EDTA, about 1 mg / mL polysorbate 80, and the pH of the antibody formulation is 5.9±0.3.

[0053] In some embodiments, the antibody formulation comprises about 175 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, about 110 mM arginine hydrochloride, about 44 mM sucrose, about 10 mM methionine, about 1 mg / mL polysorbate 80, and the pH of the antibody formulation is 5.9±0.3.

[0054] In some embodiments, the antibody formulation comprises about 175 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, about 20 mM citric acid, about 110 mM arginine hydrochloride, about 44 mM sucrose, and about 1 mg / mL polysorbate 80, and the pH of the antibody formulation is 5.9±0.3.

[0055] In some embodiments, the antibody formulation comprises about 175 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, 33±5 mM acetic acid, about 217 mM proline, about 10 mM methionine, about 1 mg / mL polysorbate 80, and the pH of the antibody formulation is 5.9±0.3.

[0056] In some embodiments, the antibody formulation comprises about 175 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, about 110 mM arginine hydrochloride, about 44 mM sucrose, and about 2 mg / mL polysorbate 80, and the pH of the antibody formulation is 5.9±0.3.

[0057] In some embodiments, the antibody preparation comprises about 175 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, about 110 mM arginine hydrochloride, about 44 mM sucrose, about 0.5 mg / mL poloxamer 188 (equivalent to about 0.05% poloxamer 188), and the pH of the antibody preparation is 5.9±0.3.

[0058] In some embodiments, the antibody formulation comprises about 175 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, about 217 mM proline, about 2 mg / mL polysorbate 80, and the pH of the antibody formulation is 5.9±0.3.

[0059] In some embodiments, the antibody formulation comprises about 175 mg / mL anti-IL-4Rα antibody, about 10 mM histidine, about 217 mM proline, about 2 mg / mL polysorbate 80, and the pH of the antibody formulation is 5.9±0.3.

[0060] In some embodiments, the antibody formulation comprises about 175 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, 31±5 mM acetic acid, about 217 mM proline, about 2 mg / mL polysorbate 80, and the pH of the antibody formulation is 5.9±0.3.

[0061] In some embodiments, the antibody formulation comprises about 175 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, 26±5 mM acetic acid, about 217 mM proline, about 2 mg / mL polysorbate 80, and the pH of the antibody formulation is 5.9±0.3.

[0062] In some embodiments, the antibody formulation comprises about 150 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, 26±5 mM acetic acid, about 217 mM proline, about 2 mg / mL polysorbate 80, and the pH of the antibody formulation is 5.9±0.3.

[0063] In some embodiments, the antibody formulation comprises about 175 mg / mL anti-IL-4Rα antibody, about 10 mM histidine, 20±5 mM acetic acid, about 191 mM proline, about 2 mg / mL polysorbate 80, and the pH of the antibody formulation is 5.9±0.3.

[0064] In some embodiments, the antibody formulation comprises about 150 mg / mL anti-IL-4Rα antibody, about 10 mM histidine, 20±5 mM acetic acid, about 191 mM proline, about 2 mg / mL polysorbate 80, and the pH of the antibody formulation is 5.9±0.3.

[0065] In some embodiments, the antibody formulation comprises about 175 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, about 217 mM proline, about 10 mM methionine, about 2 mg / mL polysorbate 80, and the pH of the antibody formulation is 5.9±0.3.

[0066] In some embodiments, the antibody formulation comprises about 175 mg / mL anti-IL-4Rα antibody, about 10 mM histidine, about 217 mM proline, about 10 mM methionine, about 2 mg / mL polysorbate 80, and the pH of the antibody formulation is 5.9±0.3.

[0067] In some embodiments, the antibody formulation comprises about 175 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, 31±5 mM acetic acid, about 217 mM proline, about 10 mM methionine, about 2 mg / mL polysorbate 80, and the pH of the antibody formulation is 5.9±0.3.

[0068] In some embodiments, the antibody formulation comprises about 175 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, 26±5 mM acetic acid, about 217 mM proline, about 10 mM methionine, about 2 mg / mL polysorbate 80, and the pH of the antibody formulation is 5.9±0.3.

[0069] In some embodiments, the antibody formulation comprises about 150 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, 26±5 mM acetic acid, about 217 mM proline, about 10 mM methionine, about 2 mg / mL polysorbate 80, and the pH of the antibody formulation is 5.9±0.3.

[0070] In some embodiments, the antibody formulation comprises about 175 mg / mL anti-IL-4Rα antibody, about 10 mM histidine, 20±5 mM acetic acid, about 191 mM proline, about 10 mM methionine, about 2 mg / mL polysorbate 80, and the pH of the antibody formulation is 5.9±0.3.

[0071] In some embodiments, the antibody formulation comprises about 150 mg / mL anti-IL-4Rα antibody, about 10 mM histidine, 20±5 mM acetic acid, about 191 mM proline, about 10 mM methionine, about 2 mg / mL polysorbate 80, and the pH of the antibody formulation is 5.9±0.3.

[0072] In some embodiments, the anti-IL-4Rα antibody comprises a heavy chain variable region shown in SEQ ID NO:1 and a light chain variable region shown in SEQ ID NO:2.

[0073] In some embodiments, the heavy chain of the anti-IL-4Rα antibody comprises the sequence as shown in SEQ ID NO: 3, or a sequence having at least 90% identity with the sequence shown in SEQ ID NO: 3, or an amino acid sequence having one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO: 3; and / or

[0074] The light chain of the anti-IL-4Rα antibody comprises the sequence shown in SEQ ID NO:4, or a sequence having at least 90% identity with the sequence shown in SEQ ID NO:4, or an amino acid sequence having one or more conservative amino acid substitutions compared with the sequence shown in SEQ ID NO:4.

[0075] In some embodiments, the heavy chain of the anti-IL-4Rα antibody comprises the sequence shown in SEQ ID NO:3, and the light chain of the anti-IL-4Rα antibody comprises the sequence shown in SEQ ID NO:4.

[0076] In some embodiments, the anti-IL-4Rα antibody is Dupilumab (eg, Dupilumab or their biosimilars).

[0077] In some embodiments, the antibody formulation is a liquid formulation.

[0078] In some embodiments, the liquid formulation contains water.

[0079] In some embodiments, the present invention provides a method for preparing the antibody preparation, comprising: taking the prescribed amount of each component, adding water to dissolve and mix, and adjusting the pH value to 5.0-7.0 to obtain the antibody preparation.

[0080] In some embodiments, the present invention provides a method for preparing the antibody preparation, comprising: preparing a buffer solution, ultrafiltering the anti-IL-4Rα antibody into a buffer solution, adding excipients, and diluting the antibody to a specified concentration to obtain an antibody preparation.

[0081] In some embodiments, the antibody preparation is a lyophilized preparation obtained by freeze-drying the antibody preparation described herein.

[0082] In some embodiments, the antibody formulation is a reconstituted formulation obtained by reconstituting a lyophilized formulation described herein.

[0083] In some embodiments, the present invention provides the use of the antibody formulations described herein in the preparation of a medicament for treating a disease. In some embodiments, the disease is a disease associated with IL-4R overexpression. In some embodiments, the disease is an immune disease or condition. In some embodiments, the disease is selected from asthma, nasal polyps, eczema, sinusitis, chronic sinusitis, chronic sinusitis with nasal polyps, allergic skin diseases, chronic obstructive pulmonary disease, allergic rhinitis, arthritis, inflammatory diseases, allergic reactions, autoimmune lymphoproliferative syndrome, autoimmune hemolytic anemia, Barrett's esophagus, autoimmune uveitis, tuberculosis, kidney disease, prurigo, nodular prurigo, eosinophilic esophagitis, atopic dermatitis, urticaria, chronic urticaria, pruritus, bullous pemphigoid, allergic bronchopulmonary aspergillosis, neurodermatitis, ulcerative colitis, milk allergy and peanut allergy.

[0084] In some embodiments, the present invention provides a product, which includes an antibody preparation as described herein, and a container for storing the antibody preparation. In some embodiments, the product also includes instructions for use. The container can be any container conventionally used in the art for storing drugs, such as a prefilled container (such as a prefilled syringe), an injection pen, a vial (such as a vial), a centrifuge tube, a pouch, etc. The antibody preparation of the present invention has good stability under conditions such as high temperature, room temperature, and light, and can be maintained in a liquid form for easy use.

[0085] In some embodiments, the antibody formulations of the present invention are stable for at least 2 weeks under high temperature conditions (40 ± 2 ° C). In some embodiments, the antibody formulations of the present invention are stable for at least 4 weeks under high temperature conditions (40 ± 2 ° C). In some embodiments, the antibody formulations of the present invention are stable for at least 14 days under light conditions (4500 ± 500 lx, 25 ± 2 ° C). DETAILED DESCRIPTION

[0086] The present invention will be described below by means of specific embodiments, but the contents of the present invention are not limited thereto.

[0087] It should be noted that, in the present invention, the "%" related to the preparation components refers to the weight volume (w / v) percentage. For example, 1% stabilizer in the preparation means that 100 mL of the preparation contains 1 g of stabilizer, or the stabilizer content is 0.01 g / mL.

[0088] "About" or "approximately" refers to the conventional error range of the corresponding numerical value that is easily known to those skilled in the relevant technical field. In some embodiments, "about" or "approximately" mentioned herein refers to the described numerical value and its ±10%, ±5% or ±1% range.

[0089] "Including" or "comprising" means that compositions and methods, etc. include the listed elements (such as components in a composition, steps in a method, etc.), but do not exclude other elements. When "consisting essentially of..." is used to define compositions and methods, it means excluding other elements that have a fundamental impact on the combination for the intended use, but does not exclude elements that do not substantially affect the characteristics of the composition or method. "Consisting of..." means excluding elements that are not specifically listed. Embodiments defined by each of these transition terms are within the scope of the present invention. For example, when a composition is described as including components A, B, and C, a composition consisting essentially of A, B, and C and a composition consisting of A, B, and C are independently within the scope of the present invention.

[0090] The term "buffer" is also referred to as a buffer system, which includes, but is not limited to, organic acids and salts thereof, such as succinic acid, acetic acid, citric acid, ascorbic acid, gluconic acid, carbonic acid, tartaric acid or phthalic acid and salts thereof; Tris, or inorganic acids and salts thereof, such as phosphate buffers. In addition, amino acids may also be used as buffers. Such amino acids include, but are not limited to, glycine, histidine, arginine, lysine, ornithine, isoleucine, leucine, alanine, glutamic acid or aspartic acid and salts thereof.

[0091] The amount of buffer in the present invention refers to the total amount of buffer pairs in the buffer system that constitutes the buffer. In some embodiments, molar concentration is used as the unit of the amount of the buffer, and its value refers to the molar concentration of the buffer pair in the buffer system of the buffer. For example, when the histidine buffer is composed of histidine and histidine hydrochloride, the given concentration of histidine buffer (such as 20mM) is the combined concentration of histidine and histidine hydrochloride.

[0092] The preparation of the present invention can be prepared with the auxiliary material or its hydrate. For example, histidine hydrochloride, also known as histidine hydrochloride, can be anhydrous histidine hydrochloride or a histidine hydrochloride hydrate, such as histidine hydrochloride monohydrate.

[0093] The term "surfactant" includes, but is not limited to, polysorbates (Tweens, such as polysorbate 20 and polysorbate 80); poloxamers (e.g., poloxamer 188); Triton; sodium dodecyl sulfate (SDS); sodium lauryl sulfate; sodium octyl glucoside; lauryl sulfobetaine, myristyl sulfobetaine, linoleyl sulfobetaine, or stearyl sulfobetaine; lauryl sarcosine, myristyl sarcosine, linoleyl sarcosine, or stearyl sarcosine; linoleyl betaine, myristyl betaine, or cetyl betaine ; Lauramidopropyl betaine, cocoamidopropyl betaine, linoleamidopropyl betaine, myristamidopropyl betaine, palmitopropyl betaine or isostearamidopropyl betaine (e.g. lauramidopropyl); myristamidopropyl dimethylamine, palmitopropyl dimethylamine or isostearamidopropyl dimethylamine; sodium methyl cocoyl taurate or disodium methyl oleyl taurate; polyethylene glycol, polypropylene glycol, and copolymers of ethylene and propylene glycol (e.g. Pluronics, PF68, etc.); etc. In some embodiments, the surfactant is polysorbate 80, also known as PS80, Tween 80 or Tween 80.

[0094] "Treating" a patient's disease means (1) preventing the disease from developing in a patient who is susceptible to or who has not yet developed symptoms of the disease; (2) inhibiting the disease or its symptoms or arresting its development; or (3) alleviating the disease or its symptoms or causing their regression.

[0095] "Stability" and "stable" herein refer to the fact that in a preparation containing an antibody, the antibody (including its antibody fragment) does not or only rarely aggregates, degrades or fragments under given production, preparation, transportation and / or storage conditions. A "stable" preparation maintains biological activity under given production, preparation, transportation and / or storage conditions. The stability of the antibody can be assessed by measuring the degree of aggregation, degradation or fragmentation of the preparation, such as by SEC-HPLC, IEC-HPLC, CE-SDS (NR), light inspection and turbidity, insoluble particles, DLS detection of particle size, and the like.

[0096] The active ingredient in the preparation of the present invention is an anti-IL-4Rα antibody, including but not limited to monoclonal antibodies, chimeric antibodies, dAbs (domain antibodies), single-chain antibodies, Fabs, Fab- and F(ab')2 fragments, Fvs, scFvs, and Fab expression libraries. In some embodiments, the anti-IL-4Rα antibody is obtained by cell culture and purification of cells expressing the anti-IL-4Rα antibody. In some embodiments, the amino acid sequence of the heavy chain of the anti-IL-4Rα antibody is as shown in SEQ ID NO:3, and the amino acid sequence of the light chain is as shown in SEQ ID NO:4. In some embodiments, the anti-IL-4Rα antibody is Dupilumab (Dupilumab, such as Dabitux or their biosimilars).

[0097] In the following examples, unless otherwise specified, the reagents and instruments used are conventional reagents and instruments in the art and can be obtained commercially; the methods used are conventional technical methods in the art, and those skilled in the art can undoubtedly implement the methods and obtain corresponding results based on the contents of the examples.

[0098] The anti-IL-4Rα antibody used in the following examples is Dupilumab, which includes two heavy chains with identical sequences and two light chains with identical sequences, the amino acid sequence of the heavy chain variable region is shown in SEQ ID NO: 1, the amino acid sequence of the heavy chain is shown in SEQ ID NO: 3, the amino acid sequence of the light chain variable region is shown in SEQ ID NO: 2, and the amino acid sequence of the light chain is shown in SEQ ID NO: 4.

[0099] The amino acid sequence of the heavy chain variable region is as follows:

[0100] EVQLVESGGGLEQPGGSLRLSCAGSGFTFRDYAMTWVRQAPGKGLEWVSSISGSGGNTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDRLSITIRPRYYGLDVWGQGTTVTVSS(SEQ ID NO:1)

[0101] The amino acid sequence of the heavy chain is as follows:

[0102] EVQLVESGGGLEQPGGSLRLSCAGSGFTFRDYAMTWVRQAPGKGLEWVSSISGSGGNTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDRLSITIRPRYYGLDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG (SEQ ID NO:3)

[0103] The amino acid sequence of the light chain variable region is as follows:

[0104] DIVMTQSPLSLPVTPGEPASISCRSSQSLLYSIGYNYLDWYLQKSGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGFYYCMQALQTPYTFGQGTKLEIK (SEQ ID NO:2)

[0105] The amino acid sequence of the light chain is as follows:

[0106] DIVMTQSPLSLPVTPGEPASISCRSSQSLLYSIGYNYLDWYLQKSGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGFYYCMQALQTPYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO:4)

[0107] The DNA sequences of the heavy chain and light chain of the antibody were cloned into an expression vector, and then transfected into CHO cells. After cell culture and purification, the anti-IL-4Rα antibody was obtained.

[0108] In the following examples, SEC-HPLC, IEC-HPLC and CE-SDS (NR) detection were all carried out according to conventional methods.

[0109] Example 1: pH and buffer preliminary screening experiment

[0110] Based on the mass changes of monomers, polymers, fragments, etc. of the samples under high temperature and light conditions, a buffer suitable for anti-IL-4Rα antibodies and a preliminary pH range were screened.

[0111] 1.1 Experimental methods

[0112] The ingredients and contents of each preparation are shown in Table 1. The concentrations described in the preparation design are theoretical concentrations, and the contents of each ingredient in the actual preparation can be within a certain range, such as ±10% or ±5% of the theoretical concentration. The experimental conditions and test items are shown in Table 2.

[0113] Table 1. Formulation design

[0114]

[0115] Table 2. Experimental conditions and test items

[0116]

[0117] 1.2 Experimental Results

[0118] 1.2.1 Lighting experiment

[0119] Table 3. Physical and chemical test results of samples under lighting conditions (4500±500lx, 25±2℃)

[0120]

[0121] From the data in Table 3, it can be seen that for the same buffer system, after 14 days of illumination, the lower the pH, the better the stability; at the same pH, the main peak of CE-SDS (NR) of the histidine buffer system is higher than that of citric acid, and the CE-SDS (NR) fragments and aggregates are lower than those of citric acid. Overall, the stability of the preparation containing histidine is better than that of the preparation containing citric acid.

[0122] 1.2.2 High temperature experiment

[0123] Table 4. Physical and chemical test results of samples under high temperature conditions (40±2℃)

[0124]

[0125]

[0126] According to the SEC-HPLC test results, after 4 weeks of high temperature, the antibody has good stability in both histidine and citric acid buffer systems at pH 5.6 to pH 6.5, and there is little difference between the two buffer systems.

[0127] From the IEC-HPLC test results, for the same buffer system, as the high temperature time increases, the lower the pH, the slower the main peak decreases, the slower the acid zone rises, and the better the stability. Both buffer systems have the best stability at pH 5.3-pH 5.9; at the same pH, the histidine buffer system is more stable than the citric acid buffer system.

[0128] According to the CE-SDS (NR) test results, after 4 weeks of high temperature, the stability of the antibody in the histidine buffer system of pH 5.6-pH 6.5 and the citric acid buffer system was not much different, with pH 5.9-pH 6.5 being optimal and pH 5.6 being better.

[0129] Based on the results of the comprehensive investigation of light and high temperature, light has a greater impact on the samples than high temperature. The samples should be carefully avoided from being exposed to light conditions for a long time. The histidine buffer system is more stable than the citric acid buffer system as a whole. Based on comprehensive analysis, the buffer solution is tentatively determined to be histidine buffer with a pH range of 5.6 to 6.2.

[0130] Example 2: Stabilizer screening experiment

[0131] Based on 20 mM histidine buffer at pH 5.9, stabilizers were screened to understand the quality change trend of anti-IL-4Rα antibodies under different formulation component conditions and to determine stabilizers with good protective effects.

[0132] 1.1 Experimental methods

[0133] The ingredients and contents of each preparation are shown in Table 5. The experimental conditions and test items are shown in Table 6.

[0134] Table 5. Formulation design

[0135]

[0136] Table 6. Experimental conditions and test items

[0137]

[0138]

[0139] 1.2 Experimental Results

[0140] 1.2.1 Lighting experiment

[0141] Table 7. Physical and chemical test results of samples under lighting conditions (4500±500lx, 25±2℃)

[0142]

[0143] As can be seen from Table 7, after 12 days of illumination, the SEC monomers of preparations 2-6 and 2-7 were the highest, the aggregates were the least, and the stability was relatively the best, preparations 2-3 and 2-9 were relatively good, and preparations 2-2 and 2-5 were relatively poor. After 12 days of illumination, the main peaks of CE-SDS (NR) of each preparation were not much different.

[0144] 1.2.2 High temperature experiment

[0145] Table 8. Physical and chemical test results of samples under high temperature conditions (40±2℃)

[0146]

[0147] From the data in Table 8, it can be seen that after 4 weeks of high temperature experiment, the SEC test results of each preparation are not much different. Compared with other preparations, the SEC aggregate of the proline preparation rises the slowest, showing better stability. After 4 weeks of high temperature, the main peak of IEC of preparation 2-4 without stabilizer, preparation 2-5 with mannitol, and preparation 2-9 with proline is slightly higher than that of other preparations, and the acid peak is slightly lower, and the stability is better. After 4 weeks of high temperature, the main peak of CE-SDS (NR) of preparation 2-6 with arginine hydrochloride added is the lowest, the fragment is the highest, and the stability is the worst. The CE-SDS (NR) test results of the other preparations are not much different.

[0148] Based on the results of protein stability analysis, light exposure and high temperature investigation, trehalose and mannitol formulations had the worst stability; single arginine hydrochloride had poor stability, arginine hydrochloride and sucrose had good stability when added at the same time, and proline preparations had good stability as a whole.

[0149] Example 3: PS80 degradation study

[0150] The effects of different buffer systems, different antioxidants and metal chelators on the degradation of PS80 in the preparation were investigated.

[0151] 1.1 Experimental methods

[0152] The ingredients and contents of each preparation are shown in Table 9. The experimental conditions and test items are shown in Table 10.

[0153] Table 9. Formulation design

[0154]

[0155] Table 10. Experimental conditions and test items

[0156]

[0157] 1.2 Experimental Results

[0158] 1.2.1 Antibody concentration, pH and osmotic pressure analysis results

[0159] Table 11. Antibody concentration, pH and osmotic pressure analysis results

[0160]

[0161] It can be seen from the data in Table 11 that formulations 3-4, 3-5 and control formulation 3-6 all have relatively high osmotic pressures.

[0162] 1.2.2 Lighting experiment

[0163] Table 12. Physical and chemical test results of samples under lighting conditions (4500±500lx, 25±2℃)

[0164]

[0165] As can be seen from Table 12, after 12 days of illumination, there was no significant difference in the SEC test results of the preparation samples, and there was little difference in the CE-SDS (NR) test results of the preparations.

[0166] Table 13. Results of polysorbate 80 content test under illumination conditions (4500±500lx, 25±2℃)

[0167]

[0168]

[0169] It can be seen from the data in Table 13 that after 12 days of illumination, the PS80 content of preparation 3-5 decreased by about 15%, indicating that preparation 3-5 can effectively slow down the degradation of PS80 in the preparation under illumination conditions.

[0170] 1.2.3 High temperature experiment

[0171] Table 14. Physical and chemical test results of samples under high temperature conditions (40±2℃)

[0172]

[0173] From the data in Table 14, it can be seen that after 4 weeks of high temperature experiment, there was no significant difference in the SEC test results of each preparation sample. After 4 weeks of high temperature, the IEC main peak of preparation 3-4 with added citric acid decreased relatively quickly, the acid area increased rapidly, and the stability was relatively poor. There was no significant difference in the IEC test results of the other preparation samples. After 4 weeks of high temperature, the CE-SDS (NR) test results of each preparation were not much different.

[0174] Table 15. Results of polysorbate 80 content test under high temperature conditions (40±2℃)

[0175]

[0176] From the data in Table 15, it can be seen that formulations 3-5 can better slow down the degradation of PS80 in the formulations under high temperature conditions compared with other formulations.

[0177] In summary, the sample quality of the preparations with citric acid added decreased rapidly under light and high temperature examinations, and there was no obvious difference in the physical and chemical quality of the other preparations. However, the preparation 3-5PS80 using proline as a stabilizer degraded the slowest.

[0178] Example 4: Preparation stability study

[0179] The effects of different buffers, stabilizers and surfactants on the stability of the preparation were investigated.

[0180] 1.1 Experimental methods

[0181] The ingredients and contents of each preparation are shown in Table 16. The experimental conditions and test items are shown in Table 17.

[0182] Table 16. Formulation design

[0183]

[0184] Table 17. Experimental conditions and test items

[0185]

[0186] 1.2 Experimental Results

[0187] 1.2.1 Viscosity and sliding force test results

[0188] Table 18. Viscosity and sliding force test results of formulation samples

[0189]

[0190] Note: When testing the sliding force, the inner packaging material is: 2.25mL pre-filled syringe (with 27G1 / 2 injection needle), the filling volume is: 1.14mL / piece

[0191] From the data in Table 18, it can be seen that the viscosity of preparations 4-1 and 4-2 is lower than that of other preparations, indicating that the viscosity reduction effect of arginine hydrochloride is better than that of proline. The sliding force of preparations 4-1 and 4-2 after filling prefilled syringes (PFS) is also lower than that of proline preparations. Compared with preparations 4-5 and 4-6, the viscosity is slightly higher, but the sliding force is not much different. The viscosity and sliding force of each preparation in the prefilled syringe are within an acceptable range.

[0192] 1.2.2 Antibody concentration, pH and osmotic pressure analysis results

[0193] Table 19. Antibody concentration, pH and osmotic pressure analysis results

[0194]

[0195] 1.2.3 Lighting experiment

[0196] Table 20. Physical and chemical test results of samples under lighting conditions (4500±500lx, 25±2℃)

[0197]

[0198]

[0199] From the data in Table 20, it can be seen that after 14 days of illumination, the SEC aggregates of preparation 4-4 increased rapidly, and the SEC test results of the other preparation samples had no significant difference, indicating that the samples with higher buffer concentrations had better stability. After 14 days of illumination, the CE-SDS (NR) fragments of preparation 4-1 increased slightly faster, and the test results of the other preparations had little difference.

[0200] Table 21. Surfactant content test results under light conditions (4500±500lx, 25±2℃)

[0201]

[0202] It can be seen from the data in Table 21 that under light conditions, proline and methionine can inhibit the degradation of PS80 more than arginine hydrochloride; and poloxamer 188 in preparation 4-2 is relatively stable and not degraded.

[0203] 1.2.4 High temperature experiment

[0204] Table 22. Physical and chemical test results of samples under high temperature conditions (40±2℃)

[0205]

[0206]

[0207] From the data in Table 22, it can be seen that after 4 weeks of high temperature experiment, the monomer of preparation 4-1SEC decreased significantly, the aggregate increased significantly, and the stability was poor. The other preparations did not differ much. After 4 weeks of high temperature, the main peak of preparation 4-1IEC decreased significantly, the acid region increased significantly, and the stability was relatively poor. There was no significant difference in the IEC test results of the other preparations. After 4 weeks of high temperature, the main peak of preparation 4-1CE-SDS (NR) decreased rapidly, the fragment increased rapidly, and the stability was poor. The other preparations did not differ much.

[0208] Table 23. Surfactant content test results under high temperature conditions (40±2℃)

[0209]

[0210] It can be seen from the data in Table 23 that under high temperature conditions, proline and methionine can also inhibit the degradation of PS80 better than arginine hydrochloride; poloxamer 188 in preparation 4-2 is relatively stable and not degraded.

[0211] 1.2.5 Acceleration Experiment

[0212] Table 24. Physical and chemical test results of samples under accelerated conditions (25±2℃)

[0213]

[0214]

[0215] From the data in Table 24, it can be seen that after 3 months of accelerated experiments, the SEC monomer purity of each preparation was above 98.5%, showing good stability, and there was no significant difference between the preparations. After 3 months of acceleration, the main peak of IEC of preparations 4-4, 4-5 and 4-6 decreased and the acid region increased more slowly. After 3 months of acceleration, the main peak of CE-SDS (NR) of preparations 4-4, 4-5 and 4-6 decreased and the fragment increased more slowly.

[0216] Table 25. Surfactant content test results under accelerated conditions (25±2℃)

[0217]

[0218] From the data in Table 25, it can be seen that under high temperature accelerated conditions, proline and methionine can also inhibit the degradation of PS80 better than arginine hydrochloride; poloxamer 188 in preparation 4-2 is relatively stable and not degraded.

[0219] In summary, the viscosity of preparation 4-1 is the lowest compared to other preparations, but its stability is poor under various conditions. Proline and methionine are beneficial to the stability of PS80, and poloxamer 188 shows good stability under various conditions.

[0220] Example 5: Preparation stability study

[0221] The stability of two protein concentration preparations, 175 mg / mL and 150 mg / mL, was investigated under different buffer concentrations and proline concentrations, the range of variation of viscosity and osmotic pressure was explored, and the stability of the preparation was verified.

[0222] 1.1 Experimental methods

[0223] The ingredients and contents of each preparation are shown in Table 26. The experimental conditions and test items are shown in Table 27.

[0224] Table 26. Formulation design

[0225]

[0226] Table 27. Experimental conditions and test items

[0227]

[0228] 1.2 Experimental Results

[0229] 1.2.1 Antibody concentration, pH and osmotic pressure test results

[0230] Table 28. Antibody concentration, pH and osmotic pressure test results

[0231] Preparation No. Protein concentration (mg / mL) pH Osmol / kg 5-1 174.29 5.94 295 5-2 151.14 5.92 286 5-3 174.36 5.89 356 5-4 150.50 5.90 343

[0232] From the data in Table 28, it can be seen that for preparations with the same excipients, the osmotic pressure of preparations with a protein concentration of 150 mg / mL will be approximately 10 mOsmol / kg lower than that of preparations with a protein concentration of 175 mg / mL. For preparations with the same protein concentration, when the buffer concentration is reduced by 10 mM, the proline concentration is reduced by 0.3%, and the osmotic pressure decreases by approximately 60 mOsmol / kg.

[0233] 1.2.2 Viscosity and sliding force test results

[0234] Table 29. Viscosity and sliding force test results

[0235]

[0236]

[0237] From the data in Table 29, it can be seen that for the same protein concentration, the viscosity of the sample will increase by about 1.5 cP when the buffer concentration decreases by 10 mM and the proline concentration decreases by 0.3%. For the preparations with the same excipients, the viscosity of the preparations with a protein concentration of 150 mg / mL is about 6 cP lower than that of 175 mg / mL. For the preparations with a protein concentration of 175 mg / mL, the sliding force of the 1 mL slender prefilled syringe (with a 27G1 / 2 injection needle) is about 50% lower than that of the 2.25 mL prefilled syringe (with a 27G1 / 2 injection needle). The viscosity of each preparation and the sliding force in the prefilled syringe are within the acceptable range.

[0238] 1.2.3 Lighting experiment

[0239] Table 30. Physical and chemical test results of samples under lighting conditions (4500±500lx, 25±2℃)

[0240]

[0241]

[0242] It can be seen from Table 30 that after 14 days of illumination, the SEC monomers of preparations 5-1 and 5-2 decreased to a similar extent, and the SEC monomers of preparations 5-3 and 5-4 decreased to a similar extent. Preparations 5-3 and 5-4, i.e., preparations with high buffer concentration and high proline content, had better stability. There was no significant difference in the SEC test results between samples of the same preparation with different protein concentrations. After 14 days of illumination, the main peak decline trend and amplitude of CE-SDS (NR) of the four preparations with different excipient concentrations and different protein concentrations were basically the same, with no significant difference.

[0243] Table 31. Results of polysorbate 80 content test under illumination conditions (4500±500lx, 25±2°C)

[0244]

[0245] From the data in Table 31, it can be seen that after 14 days of illumination, the PS80 content of the four preparations with different excipient concentrations and different protein concentrations decreased by about 5%, which was relatively stable overall, and there was no significant difference between the preparations.

[0246] 1.2.4 High temperature experiment

[0247] Table 32. Physical and chemical test results of samples under high temperature conditions (40±2℃)

[0248]

[0249] From the data in Table 32, it can be seen that after 4 weeks of high temperature, the downward trends and amplitudes of the SEC main peak, IEC main peak, and CE-SDS (NR) main peak of the four preparations with different excipient concentrations and different protein concentrations were basically consistent, with no significant differences, and the overall stability was good.

[0250] Table 33. Results of polysorbate 80 content test under high temperature conditions (40±2°C)

[0251]

[0252] From the data in Table 33, it can be seen that after 4 weeks of high temperature, the PS80 content of the four preparations with different excipient concentrations and different protein concentrations decreased by about 30%, and there was no significant difference between the preparations.

[0253] Based on the above results, the stability of the two protein concentrations of 150 mg / mL and 175 mg / mL was good, with no significant difference. Under light conditions, the stability of preparations 5-3 and 5-4 was slightly better than that of preparations 5-1 and 5-2, but the difference was not significant.

Claims

1. An antibody preparation comprising 10-200 mg / mL of an anti-IL-4Rα antibody, a buffer, a stabilizer and a surfactant; the anti-IL-4Rα antibody comprises a heavy chain variable region shown in SEQ ID NO: 1 and a light chain variable region shown in SEQ ID NO: 2; the stabilizer is selected from a combination of glycine, proline, and proline and methionine; the pH of the antibody preparation is 5.0-7.0, or 5.3-6.5, or 5.9±0.

3.

2. The antibody formulation of claim 1, wherein the concentration of the stabilizer is 50-500 mM, or 100-450 mM, or 120-350 mM, or 145-330 mM.

3. The antibody formulation of claim 1 or 2, wherein the stabilizer comprises 100-300 mM proline; or, the stabilizer comprises 180-230 mM proline; or, the stabilizer comprises about 217 mM proline; or, the stabilizer comprises about 191 mM proline.

4. An antibody preparation comprising 10-200 mg / mL of an anti-IL-4Rα antibody, as well as a buffer, a stabilizer and a surfactant; the anti-IL-4Rα antibody comprises a heavy chain variable region shown in SEQ ID NO:1 and a light chain variable region shown in SEQ ID NO:2; the stabilizer comprises 20-100 mM sucrose and 101-300 mM arginine or a pharmaceutically acceptable salt thereof; the pH of the antibody preparation is 5.0-7.0, or 5.3-6.5, or 5.9±0.

3.

5. The antibody formulation of any one of claims 1 to 4, wherein the buffer is selected from the group consisting of histidine, citric acid, histidine-acetate and histidine-citrate.

6. The antibody formulation of any one of claims 1-5, wherein the concentration of the buffer is 1-80 mM, or 5-80 mM, or 10-80 mM, or 10-60 mM, or 15-60 mM, or 20-55 mM.

7. The antibody preparation of any one of claims 1-6, wherein the concentration of the anti-IL-4Rα antibody is 20-200 mg / mL, or 40-200 mg / mL, or 20-150 mg / mL, or 100-200 mg / mL, or 120-180 mg / mL; or 145-180 mg / mL, or 20-80 mg / mL.

8. The antibody preparation according to any one of claims 2 to 7, further comprising 1-30 mM methionine or 0.01-0.5 mM EDTA. 9 . The antibody preparation according to claim 1 , wherein the surfactant is polysorbate or poloxamer; or, the surfactant is polysorbate 80, polysorbate 20 or poloxamer 188.

10. The antibody preparation according to any one of claims 1 to 9, wherein the concentration of the surfactant is 0.1-5 mg / mL, or 0.5-5 mg / mL, or 0.5-3 mg / mL.

11. An antibody preparation, wherein the antibody preparation comprises 20-200 mg / mL anti-IL-4Rα antibody, 5-80 mM buffer, 100-450 mM stabilizer, 0.1-5 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody preparation is 5.3-6.5; or The antibody preparation comprises 20-100 mg / mL anti-IL-4Rα antibody, 10-30 mM buffer, 120-350 mM stabilizer, and 0.5-3 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody preparation is 5.3-6.5; or The antibody preparation comprises 45-55 mg / mL anti-IL-4Rα antibody, 10-30 mM buffer, 145-330 mM stabilizer, and 1-3 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody preparation is 5.3-6.5; or The antibody preparation comprises 100-200 mg / mL anti-IL-4Rα antibody, 5-80 mM buffer, 120-350 mM stabilizer, and 0.5-3 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody preparation is 5.3-6.5; or The antibody preparation comprises 130-200 mg / mL anti-IL-4Rα antibody, 10-60 mM buffer, 145-330 mM stabilizer, and 0.5-3 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody preparation is 5.3-6.5; wherein the buffer is selected from the group consisting of histidine, citric acid, histidine-acetic acid and histidine-citrate; The stabilizer is selected from the group consisting of glycine, proline, and a combination of proline and methionine; The anti-IL-4Rα antibody comprises a heavy chain variable region shown in SEQ ID NO: 1 and a light chain variable region shown in SEQ ID NO:

2.

12. An antibody preparation, wherein the antibody preparation comprises 20-200 mg / mL anti-IL-4Rα antibody, 5-80 mM buffer, 20-100 mM sucrose, 101-300 mM arginine or a pharmaceutically acceptable salt thereof, 0.1-5 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody preparation is 5.3-6.5; or The antibody preparation comprises 20-200 mg / mL anti-IL-4Rα antibody, 5-80 mM buffer, 20-100 mM sucrose, 101-300 mM arginine hydrochloride, and 0.5-3 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody preparation is 5.3-6.5; or The antibody preparation comprises 45-55 mg / mL anti-IL-4Rα antibody, 10-30 mM buffer, 35-90 mM sucrose, 101-245 mM arginine hydrochloride, and 1-3 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody preparation is 5.3-6.5; or The antibody preparation comprises 100-200 mg / mL anti-IL-4Rα antibody, 5-80 mM buffer, 35-90 mM sucrose, 101-245 mM arginine hydrochloride, and 0.5-3 mg / mL polysorbate 80 or poloxamer 188, and the pH of the antibody preparation is 5.3-6.5; wherein the buffer is selected from the group consisting of histidine, citric acid, histidine-acetic acid and histidine-citrate; The anti-IL-4Rα antibody comprises a heavy chain variable region shown in SEQ ID NO: 1 and a light chain variable region shown in SEQ ID NO:

2.

13. An antibody preparation, wherein the antibody preparation comprises about 50 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, about 293 mM glycine, about 2 mg / mL polysorbate 80, and the pH of the antibody preparation is 5.9±0.3; or The antibody preparation comprises about 50 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, about 88 mM sucrose, about 237 mM arginine hydrochloride, and about 2 mg / mL polysorbate 80, and the pH of the antibody preparation is 5.9±0.3; or The antibody preparation comprises about 50 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, about 217 mM proline, and about 2 mg / mL polysorbate 80, and the pH of the antibody preparation is 5.9±0.3; or The antibody preparation comprises about 175 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, about 110 mM arginine hydrochloride, about 44 mM sucrose, and about 1 mg / mL polysorbate 80, and the pH of the antibody preparation is 5.9±0.3; or The antibody preparation comprises about 175 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, about 110 mM arginine hydrochloride, about 44 mM sucrose, about 0.05 mM EDTA, and about 1 mg / mL polysorbate 80, and the pH of the antibody preparation is 5.9±0.3; or The antibody preparation comprises about 175 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, about 110 mM arginine hydrochloride, about 44 mM sucrose, about 10 mM methionine, and about 1 mg / mL polysorbate 80, and the pH of the antibody preparation is 5.9±0.3; or The antibody preparation comprises about 175 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, about 20 mM citric acid, about 110 mM arginine hydrochloride, about 44 mM sucrose, and about 1 mg / mL polysorbate 80, and the pH of the antibody preparation is 5.9±0.3; or The antibody preparation comprises about 175 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, 33±5 mM acetic acid, about 217 mM proline, about 10 mM methionine, and about 1 mg / mL polysorbate 80, and the pH of the antibody preparation is 5.9±0.3; or The antibody preparation comprises about 175 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, about 110 mM arginine hydrochloride, about 44 mM sucrose, and about 2 mg / mL polysorbate 80, and the pH of the antibody preparation is 5.9±0.3; or The antibody preparation comprises about 175 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, about 110 mM arginine hydrochloride, about 44 mM sucrose, and about 0.5 mg / mL poloxamer 188, and the pH of the antibody preparation is 5.9±0.3; or The antibody preparation comprises about 175 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, about 217 mM proline, about 10 mM methionine, and about 2 mg / mL polysorbate 80, and the pH of the antibody preparation is 5.9±0.3; or The antibody preparation comprises about 175 mg / mL anti-IL-4Rα antibody, about 10 mM histidine, about 217 mM proline, about 10 mM methionine, and about 2 mg / mL polysorbate 80, and the pH of the antibody preparation is 5.9±0.3; or The antibody preparation comprises about 175 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, 31±5 mM acetic acid, about 217 mM proline, about 10 mM methionine, and about 2 mg / mL polysorbate 80, and the pH of the antibody preparation is 5.9±0.3; or The antibody preparation comprises about 175 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, 26±5 mM acetic acid, about 217 mM proline, about 10 mM methionine, and about 2 mg / mL polysorbate 80, and the pH of the antibody preparation is 5.9±0.3; or The antibody preparation comprises about 150 mg / mL anti-IL-4Rα antibody, about 20 mM histidine, 26±5 mM acetic acid, about 217 mM proline, about 10 mM methionine, and about 2 mg / mL polysorbate 80, and the pH of the antibody preparation is 5.9±0.3; or The antibody preparation comprises about 175 mg / mL anti-IL-4Rα antibody, about 10 mM histidine, 20±5 mM acetic acid, about 191 mM proline, about 10 mM methionine, and about 2 mg / mL polysorbate 80, and the pH of the antibody preparation is 5.9±0.3; or The antibody preparation comprises about 150 mg / mL anti-IL-4Rα antibody, about 10 mM histidine, 20±5 mM acetic acid, about 191 mM proline, about 10 mM methionine, and about 2 mg / mL polysorbate 80, and the pH of the antibody preparation is 5.9±0.3; The anti-IL-4Rα antibody comprises a heavy chain variable region shown in SEQ ID NO: 1 and a light chain variable region shown in SEQ ID NO:

2.

14. The antibody preparation of any one of claims 1 to 13, wherein the heavy chain of the anti-IL-4Rα antibody comprises the sequence as shown in SEQ ID NO: 3, or a sequence having at least 90% identity with the sequence shown in SEQ ID NO: 3, or an amino acid sequence having one or more conservative amino acid substitutions compared to the sequence shown in SEQ ID NO: 3; and / or The light chain of the anti-IL-4Rα antibody comprises the sequence shown in SEQ ID NO:4, or a sequence having at least 90% identity with the sequence shown in SEQ ID NO:4, or an amino acid sequence having one or more conservative amino acid substitutions compared with the sequence shown in SEQ ID NO:

4.

15. The antibody preparation according to any one of claims 1 to 14, wherein the anti-IL-4Rα antibody is dupilumab.

16. A method for preparing the antibody preparation according to any one of claims 1 to 15, comprising: Take the prescribed amount of each component, add water to dissolve and mix, adjust the pH value to 5.0-7.0, and obtain an antibody preparation; Alternatively, the method comprises: preparing a buffer solution, ultrafiltering the anti-IL-4Rα antibody to change the buffer solution, adding auxiliary materials, and diluting the antibody to a specified concentration to obtain an antibody preparation.

17. A lyophilized preparation obtained by freeze-drying the antibody preparation according to any one of claims 1 to 15.

18. A reconstituted preparation obtained by reconstituted the lyophilized preparation according to claim 17.

19. Use of the antibody preparation according to any one of claims 1 to 15, or the lyophilized preparation according to claim 17, or the reconstituted preparation according to claim 18 in the preparation of a medicament for treating a disease; or, the disease is a disease associated with IL-4R overexpression; or, the disease is an immune disease or condition; or, the disease is selected from asthma, nasal polyps, eczema, sinusitis, chronic sinusitis, chronic sinusitis with nasal polyps, allergic skin diseases, chronic obstructive pulmonary disease, allergic rhinitis, arthritis, inflammatory diseases, allergic reactions, autoimmune lymphoproliferative syndrome, autoimmune hemolytic anemia, Barrett's esophagus, autoimmune uveitis, tuberculosis, kidney disease, prurigo, prurigo nodularis, eosinophilic esophagitis, atopic dermatitis, urticaria, chronic urticaria, pruritus, bullous pemphigoid, allergic bronchopulmonary aspergillosis, neurodermatitis, ulcerative colitis, milk allergy and peanut allergy.

20. A product comprising the antibody preparation according to any one of claims 1 to 15, the lyophilized preparation according to claim 17, or the reconstituted preparation according to claim 18, and a container for storing the antibody preparation, lyophilized preparation, or reconstituted preparation.

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