Pharmaceutical compositions of anti-IL4R antibodies and uses thereof

Abstract

The present invention provides a pharmaceutical composition of an anti-IL4R antibody and uses thereof. The pharmaceutical composition comprises an anti-IL4R antibody or an antigen-binding fragment thereof and a buffer, and may further comprise a surfactant and a stabilizer.

Description

[Technical field]

[0001] The present disclosure is in the field of formulations, and in particular, the present disclosure relates to stable pharmaceutical compositions comprising anti-IL4R antibodies or antigen-binding fragments thereof, and uses of said pharmaceutical compositions for the treatment and / or prevention of diseases associated with excessive IL4 and / or IL13 signaling. [Background technology]

[0002] Type 2 inflammation-related allergic diseases, such as atopic dermatitis, anaphylaxis, allergic rhinitis, and allergic asthma, affect more than 3 billion people worldwide, and their incidence continues to increase. According to the hygiene hypothesis, one of the reasons for the high incidence is that as living standards improve, the opportunities for exposure to infectious substances decrease, and the immune system becomes more sensitive to allergens that are normally harmless (Stephen J. Galli et al., (2008) Nature 454 (7203): 445-454). Interleukin 4 (IL4 or IL-4) and IL-13 are two key factors in type 2 immunity and are required to drive most of the key markers associated with type 2 inflammation, such as the production of immunoglobulin E and the recruitment of innate immune cells to the site of inflammation (Gruning G et al., (1998) Science 282:2261-2263; Rankin JA et al., (1996) Proc Natl Acad Sci USA 93:7821-7825; Wills-Karp M et al., (1998) Science 282:2258-2261).

[0003] IL-4 and IL-13 regulate cell functions and activate transcriptional machinery by binding to cell surface receptors. Specifically, IL-4 first binds to the IL-4Rα chain with picomolar affinity and then recruits IL-2Rγ (γc) to form a type I receptor complex or IL-13Rα1 to form a type II receptor complex. Studies have shown that non-hematopoietic cells do not express γc or express it at low levels and highly express IL-13Rα1, whereas lymphocytes do the opposite and myeloid cells are intermediate between these two. The type II receptor complex is also formed by the binding of IL-13 to the IL-13Rα1 chain (with nanomolar affinity) and the further recruitment of the IL-4Rα chain. In addition, IL-13 can bind to IL-13Rα2 with picomolar affinity and initiate the formation of decoy receptors (Irina G. Luzina et al., (2012) J Leukoc Biol 92(4):753-764). Upon formation of the IL-4 receptor complex, intracellular signaling molecules are activated, and STAT6 and IRS signaling are activated in response to type I IL-4 receptor, whereas type II IL-4 receptor cannot significantly activate IRS (Heller NM et al., (2008) Sci Signal 1(51):ra17-ra17). STAT6 signaling is involved in the activation of T H It is very important for IL-2 cell differentiation and the production of IL-4, and IRS can activate PI3K and mTOR signaling pathways (Gadani SP et al., (2012) J Immunol 189: 4213-4219). Research has shown that excessive IL-4 and / or IL-13 signaling can cause allergic diseases. Other studies have also found that STAT6 inhibitors can inhibit the proliferation of prostate cancer cells (Nappo G et al., (2017) Oncogenesis 6 (5): e342). Therefore, therapeutic antibodies that regulate IL-4 and / or IL-13-mediated signaling have been developed one after another, one example of which is the antibody Dupilumab against IL4R.

[0004] Antibody drugs for humans need to maintain their biological activity and stability during storage and subsequent use, but antibody drugs have large molecular weights and complex structures, and are prone to instability due to degradation, polymerization, or undesirable chemical modifications. Therefore, in order to solve the above problems, the art needs to develop stable antibody pharmaceutical compositions (e.g., anti-IL4R antibody pharmaceutical compositions) that can provide antibody drugs that are suitable for production and administration to patients and have a long shelf life. However, when antibodies are present at high concentrations, problems such as high levels of aggregation, osmotic pressure higher than physiological levels, and reduced injectability due to increased viscosity, among others, need to be solved. Summary of the Invention [Problem to be solved by the invention]

[0005] The present disclosure provides pharmaceutical compositions that contain high concentrations of anti-IL4Rα antibodies or antigen-binding fragments thereof and have acceptable viscosity levels without causing high levels of aggregation. [Means for solving the problem]

[0006] The pharmaceutical composition according to the present disclosure comprises (a) an anti-IL4Rα antibody or an antigen-binding fragment thereof, (b) a buffering agent, (c) a surfactant, and (d) a stabilizer.

[0007] In some embodiments, the concentration of the anti-IL4Rα antibody or antigen-binding fragment thereof in the pharmaceutical composition is 30 mg / mL to 300 mg / mL, 50 mg / mL to 250 mg / mL, 70 mg / mL to 200 mg / mL, 100 mg / mL to 180 mg / mL, 120 mg / mL to 180 mg / mL, 120 mg / mL to 150 mg / mL, or 150 mg / mL to 180 mg / mL. In some embodiments, the concentration of the anti-IL4Rα antibody or antigen-binding fragment thereof in the pharmaceutical composition is about 150 mg / mL. In some embodiments, the concentration of the anti-IL4Rα antibody or antigen-binding fragment thereof in the pharmaceutical composition is about 175 mg / mL. In some embodiments, examples of the concentration of the anti-IL4Rα antibody or antigen-binding fragment thereof in the pharmaceutical composition include, but are not limited to, 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 125 mg / mL, about 130 mg / mL, about 135 mg / mL, about 140 mg / mL, about 145 mg / mL, about 150 mg / mL, about 155 mg / mL, about 160 mg / mL, about 165 mg / mL, about 170 mg / mL, about 175 mg / mL, about 180 mg / mL, about 190 mg / mL, or about 200 mg / mL.

[0008] In some embodiments, an example of the buffer in the pharmaceutical composition includes a phosphate buffer, an acetate buffer, or a histidine buffer. A preferred example of the buffer in the pharmaceutical composition includes a histidine buffer. In some specific embodiments, an example of the phosphate buffer includes a sodium phosphate buffer or a potassium phosphate buffer, an example of the acetate buffer includes a sodium acetate buffer, a potassium acetate buffer, or a sodium acetate-acetate buffer, and an example of the histidine buffer includes a histidine-hydrochloric acid buffer, a histidine-acetate buffer, a histidine-histidine hydrochloride buffer, or a histidine hydrochloride buffer.

[0009] In certain embodiments, the sodium phosphate buffer comprises disodium hydrogen phosphate and sodium dihydrogen phosphate. In certain embodiments, the sodium acetate-acetic acid buffer comprises sodium acetate and acetic acid. In certain embodiments, the histidine-histidine hydrochloride buffer comprises histidine and histidine hydrochloride.

[0010] In some embodiments, the concentration of the buffer in the pharmaceutical composition is 1 mM to 100 mM, 2 mM to 80 mM, 4 mM to 60 mM, 8 mM to 40 mM, 10 mM to 30 mM, 10 mM to 20 mM, or 20 mM to 30 mM. In some embodiments, the concentration of the buffer in the pharmaceutical composition is about 20 mM. In some embodiments, examples of the concentration of the buffer in the pharmaceutical composition include, but are not limited to, about 8 mM, about 9 mM, about 10 mM, about 11 mM, about 12 mM, about 13 mM, about 14 mM, about 15 mM, about 16 mM, about 17 mM, about 18 mM, about 19 mM, about 20 mM, about 21 mM, about 22 mM, about 23 mM, about 24 mM, about 25 mM, about 26 mM, about 27 mM, about 28 mM, about 30 mM, about 32 mM, about 34 mM, about 36 mM, about 38 mM, or about 40 mM.

[0011] In some embodiments, the pH of the buffer in the pharmaceutical composition is 4 to 7, 5 to 7, 5 to 6.5, 5.3 to 6.5, 5.3 to 6.3, 5.3 to 5.8, or 5.8 to 6.3. In some embodiments, the pH of the buffer in the pharmaceutical composition is about 5.8. In some embodiments, the pH of the buffer in the pharmaceutical composition is about 6. In some embodiments, the pH of the buffer in the pharmaceutical composition is about 5. In some embodiments, examples of the pH of the buffer in the pharmaceutical composition include, but are not limited to, about 5, 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, about 6.1, about 6.2, about 6.3, about 6.4, about 6.5, about 6.6, about 6.7, about 6.8, about 6.9, or about 7.

[0012] In some embodiments, the surfactant in the pharmaceutical composition is selected from a polysorbate (e.g., polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 65, polysorbate 80, polysorbate 81, or polysorbate 85), a poloxamer (e.g., poloxamer 181, poloxamer 188, or poloxamer 407), a polyethylene glycol, or a polyhydroxy substance, etc. In some embodiments, the surfactant in the pharmaceutical composition is polysorbate 80. In some embodiments, the surfactant in the pharmaceutical composition is polysorbate 20.

[0013] In some embodiments, the concentration of the surfactant in the pharmaceutical composition is 0.01 mg / mL to 2 mg / mL, 0.05 mg / mL to 1 mg / mL, 0.1 mg / mL to 0.8 mg / mL, 0.2 mg / mL to 0.6 mg / mL, or 0.2 mg / mL to 0.4 mg / mL. In some embodiments, the concentration of the surfactant in the pharmaceutical composition is about 0.4 mg / mL. In some embodiments, examples of the concentration of the surfactant in the pharmaceutical composition include, but are not limited to, about 0.05 mg / mL, about 0.06 mg / mL, about 0.07 mg / mL, about 0.08 mg / mL, about 0.09 mg / mL, about 0.1 mg / mL, about 0.2 mg / mL, about 0.3 mg / mL, about 0.4 mg / mL, about 0.5 mg / mL, about 0.6 mg / mL, about 0.7 mg / mL, about 0.8 mg / mL, about 0.9 mg / mL, or about 1 mg / mL.

[0014] In some embodiments, the stabilizer in the pharmaceutical composition is one or more selected from trehalose, mannitol, sucrose, arginine or a pharma- ceutically acceptable salt thereof, proline, or sodium chloride. In some embodiments, the stabilizer in the pharmaceutical composition comprises sucrose. In some embodiments, the stabilizer in the pharmaceutical composition comprises arginine or a pharma- ceutically acceptable salt thereof. In some embodiments, the stabilizer in the pharmaceutical composition comprises proline. In some embodiments, the stabilizer in the pharmaceutical composition comprises sodium chloride. An example of a pharma- ceutically acceptable salt of arginine is arginine hydrochloride or arginine acetate. In some particular embodiments, the stabilizer in the pharmaceutical composition comprises sucrose and arginine hydrochloride. In some particular embodiments, the stabilizer in the pharmaceutical composition comprises proline and sodium chloride. In some particular embodiments, the stabilizer in the pharmaceutical composition comprises sucrose, proline, and sodium chloride.

[0015] In some embodiments, the concentration of sucrose in the pharmaceutical composition is 10 mg / mL to 100 mg / mL, 20 mg / mL to 80 mg / mL, 30 mg / mL to 60 mg / mL, 40 mg / mL to 50 mg / mL, or 50 mg / mL to 60 mg / mL. In some embodiments, the concentration of sucrose in the pharmaceutical composition is about 50 mg / mL. In some embodiments, examples of the concentration of sucrose in the pharmaceutical composition include, but are not limited to, about 20 mg / mL, about 22 mg / mL, about 24 mg / mL, about 26 mg / mL, about 28 mg / mL, about 30 mg / mL, about 32 mg / mL, about 34 mg / mL, about 36 mg / mL, about 38 mg / mL, about 40 mg / mL, about 42 mg / mL, about 44 mg / mL, about 46 mg / mL, about 48 mg / mL, about 50 mg / mL, about 52 mg / mL, about 54 mg / mL, about 56 mg / mL, about 58 mg / mL, about 60 mg / mL, about 62 mg / mL, about 64 mg / mL, about 66 mg / mL, about 68 mg / mL, about 70 mg / mL, about 72 mg / mL, about 74 mg / mL, about 76 mg / mL, about 78 mg / mL, or about 80 mg / mL.

[0016] In some embodiments, the concentration of the arginine or a pharma- ceutically acceptable salt thereof (e.g., arginine hydrochloride) in the pharmaceutical composition is 10 mM to 100 mM, 20 mM to 80 mM, 30 mM to 60 mM, 40 mM to 50 mM, or 50 mM to 60 mM. In some embodiments, the concentration of the arginine or a pharma- ceutically acceptable salt thereof (e.g., arginine hydrochloride) in the pharmaceutical composition is about 50 mM. In some embodiments, examples of the concentration of arginine or a pharma- ceutically acceptable salt thereof (e.g., arginine hydrochloride) in the pharmaceutical composition include, but are not limited to, about 20 mM, about 22 mM, about 24 mM, about 26 mM, about 28 mM, about 30 mM, about 32 mM, about 34 mM, about 36 mM, about 38 mM, about 40 mM, about 42 mM, about 44 mM, about 46 mM, about 48 mM, about 50 mM, about 52 mM, about 54 mM, about 56 mM, about 58 mM, about 60 mM, about 62 mM, about 64 mM, about 66 mM, about 68 mM, about 70 mM, about 72 mM, about 74 mM, about 76 mM, about 78 mM, or about 80 mM.

[0017] In some embodiments, the concentration of the proline in the pharmaceutical composition is 60 mM to 600 mM, 80 mM to 500 mM, 100 mM to 450 mM, 120 mM to 400 mM, 150 mM to 350 mM, 200 mM to 350 mM, or 250 mM to 350 mM. In some embodiments, the concentration of the proline in the pharmaceutical composition is about 250 mM. In some embodiments, the concentration of the proline in the pharmaceutical composition is about 350 mM. In some embodiments, examples of the concentration of proline in the pharmaceutical composition include, but are not limited to, about 120 mM, about 130 mM, about 140 mM, about 150 mM, about 160 mM, about 170 mM, about 180 mM, about 190 mM, about 200 mM, about 210 mM, about 220 mM, about 230 mM, about 240 mM, about 250 mM, about 260 mM, about 270 mM, about 280 mM, about 290 mM, about 300 mM, about 310 mM, about 320 mM, about 330 mM, about 340 mM, about 350 mM, about 360 mM, about 370 mM, about 380 mM, about 390 mM, or about 400 mM.

[0018] In some embodiments, the concentration of the sodium chloride in the pharmaceutical composition is 1 mM to 80 mM, 2 mM to 60 mM, 4 mM to 50 mM, 8 mM to 40 mM, 10 mM to 30 mM, or 10 mM to 20 mM. In some embodiments, the concentration of the sodium chloride in the pharmaceutical composition is about 20 mM. In some embodiments, examples of the concentration of sodium chloride in the pharmaceutical composition include, but are not limited to, about 8 mM, about 9 mM, about 10 mM, about 11 mM, about 12 mM, about 13 mM, about 14 mM, about 15 mM, about 16 mM, about 17 mM, about 18 mM, about 19 mM, about 20 mM, about 21 mM, about 22 mM, about 23 mM, about 24 mM, about 25 mM, about 26 mM, about 27 mM, about 28 mM, about 29 mM, about 30 mM, about 32 mM, about 34 mM, about 36 mM, about 38 mM, or about 40 mM.

[0019] In some embodiments, the pH of the pharmaceutical composition is 4 to 7, 5 to 7, 5 to 6.5, 5.3 to 6.5, 5.3 to 6.3, 5.3 to 5.8, or 5.8 to 6.3. In some embodiments, the pH of the pharmaceutical composition is about 5.8. In some embodiments, the pH of the pharmaceutical composition is about 6. In some embodiments, the pH of the pharmaceutical composition is about 5. In some embodiments, examples of the pH of the pharmaceutical composition include, but are not limited to, about 5, 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, about 6.1, about 6.2, about 6.3, about 6.4, about 6.5, about 6.6, about 6.7, about 6.8, about 6.9, or about 7.

[0020] In some embodiments, the pharmaceutical composition comprises (a) an anti-IL4Rα antibody or an antigen-binding fragment thereof; (b) a phosphate buffer, an acetate buffer, and / or a histidine buffer; (c) polysorbate 80 or polysorbate 20; and (d) trehalose, mannitol, sucrose, arginine or a pharma- ceutically acceptable salt thereof, proline, and / or sodium chloride.

[0021] In some embodiments, the pharmaceutical composition comprises (a) an anti-IL4Rα antibody or antigen-binding fragment thereof; (b) a phosphate buffer or a histidine salt buffer; (c) polysorbate 80; and (d) sucrose, arginine or a pharma- ceutically acceptable salt thereof, proline, and / or sodium chloride.

[0022] In some embodiments, the pharmaceutical composition comprises (a) an anti-IL4Rα antibody or antigen-binding fragment thereof, (b) a histidine salt buffer, (c) polysorbate 80, and (d) sucrose and arginine or a pharma- ceutically acceptable salt thereof.

[0023] In some embodiments, the pharmaceutical composition comprises (a) an anti-IL4Rα antibody or antigen-binding fragment thereof; (b) a histidine salt buffer; (c) polysorbate 80; and (d) sucrose, proline, and sodium chloride.

[0024] In some embodiments, the pharmaceutical composition comprises (a) an anti-IL4Rα antibody or antigen-binding fragment thereof, (b) a histidine salt buffer, (c) polysorbate 80, and (d) proline and sodium chloride.

[0025] In some embodiments, the pharmaceutical composition comprises (a) an anti-IL4Rα antibody or antigen-binding fragment thereof, (b) a histidine salt buffer, (c) polysorbate 80, and (d) proline.

[0026] In some embodiments, the pharmaceutical composition comprises (a) 30 mg / mL to 300 mg / mL of an anti-IL4Rα antibody or an antigen-binding fragment thereof, (b) 1 mM to 100 mM of a buffering agent, (c) 0.01 mg / mL to 2 mg / mL of a surfactant, and (d) a stabilizer comprising trehalose, mannitol, sucrose, arginine or a pharma- ceutically acceptable salt thereof, proline and / or sodium chloride, and the pH of the pharmaceutical composition is 4 to 7, 5 to 7, 5 to 6.5, 5.3 to 6.5, 5.3 to 6.3, 5.3 to 5.8, or 5.8 to 6.3.

[0027] In some embodiments, the pharmaceutical composition comprises (a) 50 mg / mL to 250 mg / mL of an anti-IL4Rα antibody or an antigen-binding fragment thereof, (b) 2 mM to 80 mM of a buffering agent, (c) 0.05 mg / mL to 1 mg / mL of a surfactant, and (d) a stabilizer comprising trehalose, mannitol, sucrose, arginine or a pharma- ceutically acceptable salt thereof, proline and / or sodium chloride, and the pH of the pharmaceutical composition is 4 to 7, 5 to 7, 5 to 6.5, 5.3 to 6.5, 5.3 to 6.3, 5.3 to 5.8, or 5.8 to 6.3.

[0028] In some embodiments, the pharmaceutical composition comprises (a) 70 mg / mL to 200 mg / mL of an anti-IL4Rα antibody or an antigen-binding fragment thereof, (b) 4 mM to 60 mM of a buffering agent, (c) 0.1 mg / mL to 0.8 mg / mL of a surfactant, and (d) a stabilizer comprising trehalose, mannitol, sucrose, arginine or a pharma- ceutically acceptable salt thereof, proline and / or sodium chloride, and the pH of the pharmaceutical composition is 4 to 7, 5 to 7, 5 to 6.5, 5.3 to 6.5, 5.3 to 6.3, 5.3 to 5.8, or 5.8 to 6.3.

[0029] In some embodiments, the pharmaceutical composition comprises: (a) 100 mg / mL to 180 mg / mL or 120 mg / mL to 180 mg / mL of an anti-IL4Rα antibody or antigen-binding fragment thereof; (b) 8 mM to 40 mM or 10 mM to 30 mM of a buffer; (c) 0.2 mg / mL to 0.6 mg / mL of a surfactant; and (d) a stabilizer comprising trehalose, mannitol, sucrose, arginine or a pharma- ceutically acceptable salt thereof, proline and / or sodium chloride; and the pH of the pharmaceutical composition is 4 to 7, 5 to 7, 5 to 6.5, 5.3 to 6.5, 5.3 to 6.3, 5.3 to 5.8, or 5.8 to 6.3.

[0030] In some embodiments, the pharmaceutical composition comprises: (a) 120 mg / mL to 150 mg / mL or 150 mg / mL to 180 mg / mL of an anti-IL4Rα antibody or antigen-binding fragment thereof; (b) 10 mM to 20 mM or 20 mM to 30 mM of a buffer; (c) 0.2 mg / mL to 0.4 mg / mL of a surfactant; and (d) a stabilizer comprising trehalose, mannitol, sucrose, arginine or a pharma- ceutically acceptable salt thereof, proline and / or sodium chloride; and the pH of the pharmaceutical composition is 4 to 7, 5 to 7, 5 to 6.5, 5.3 to 6.5, 5.3 to 6.3, 5.3 to 5.8, or 5.8 to 6.3.

[0031] In some embodiments, the pharmaceutical composition comprises (a) 30 mg / mL to 300 mg / mL of an anti-IL4Rα antibody or an antigen-binding fragment thereof, (b) 1 mM to 100 mM of a phosphate buffer, acetate buffer, or histidine buffer, (c) 0.01 mg / mL to 2 mg / mL of polysorbate 80 or polysorbate 20, and (d) 10 mg / mL to 100 mg / mL of sucrose, 10 mM to 100 mM of arginine or a pharma- ceutically acceptable salt thereof, 60 mM to 600 mM of proline, and / or 1 mM to 80 mM of sodium chloride, and the pH of the pharmaceutical composition is 4 to 7, 5 to 7, 5 to 6.5, 5.3 to 6.5, 5.3 to 6.3, 5.3 to 5.8, or 5.8 to 6.3.

[0032] In some embodiments, the pharmaceutical composition comprises (a) 50 mg / mL to 250 mg / mL of an anti-IL4Rα antibody or an antigen-binding fragment thereof, (b) 2 mM to 80 mM of a phosphate buffer, acetate buffer, or histidine buffer, (c) 0.05 mg / mL to 1 mg / mL of polysorbate 80 or polysorbate 20, and (d) 20 mg / mL to 80 mg / mL of sucrose, 20 mM to 80 mM of arginine or a pharma- ceutically acceptable salt thereof, 80 mM to 500 mM of proline, and / or 2 mM to 60 mM of sodium chloride, and the pH of the pharmaceutical composition is 4 to 7, 5 to 7, 5 to 6.5, 5.3 to 6.5, 5.3 to 6.3, 5.3 to 5.8, or 5.8 to 6.3.

[0033] In some embodiments, the pharmaceutical composition comprises (a) 70 mg / mL to 200 mg / mL of an anti-IL4Rα antibody or an antigen-binding fragment thereof, (b) 4 mM to 60 mM of a phosphate buffer, acetate buffer, or histidine buffer, (c) 0.1 mg / mL to 0.8 mg / mL of polysorbate 80 or polysorbate 20, and (d) 30 mg / mL to 60 mg / mL of sucrose, 30 mM to 60 mM of arginine or a pharma- ceutically acceptable salt thereof, 100 mM to 450 mM or 120 mM to 400 mM of proline, and / or 4 mM to 50 mM of sodium chloride, and the pH of the pharmaceutical composition is 4 to 7, 5 to 7, 5 to 6.5, 5.3 to 6.5, 5.3 to 6.3, 5.3 to 5.8, or 5.8 to 6.3.

[0034] In some embodiments, the pharmaceutical composition comprises: (a) 100 mg / mL to 180 mg / mL or 120 mg / mL to 180 mg / mL of an anti-IL4Rα antibody or antigen-binding fragment thereof; (b) 8 mM to 40 mM or 10 mM to 30 mM of a phosphate buffer, acetate buffer, or histidine buffer; and (c) 0.2 mg / mL to 0.6 mg / mL of polysorbate 80 or polysorb C10. and (d) 40 mg / mL to 50 mg / mL of sucrose, 40 mM to 50 mM of arginine or a pharma- ceutically acceptable salt thereof, 150 mM to 350 mM of proline, and / or 8 mM to 40 mM of sodium chloride, and the pH of the pharmaceutical composition is 4 to 7, 5 to 7, 5 to 6.5, 5.3 to 6.5, 5.3 to 6.3, 5.3 to 5.8, or 5.8 to 6.3.

[0035] In some embodiments, the pharmaceutical composition comprises: (a) 120 mg / mL to 150 mg / mL or 150 mg / mL to 180 mg / mL of an anti-IL4Rα antibody or antigen-binding fragment thereof; (b) 10 mM to 20 mM or 20 mM to 30 mM of a phosphate buffer, acetate buffer, or histidine buffer; (c) 0.2 mg / mL to 0.4 mg / mL of polysorbate 80 or polysorbate 20; and (d) 50 mM to 150 mg / mL of an anti-IL4Rα antibody or antigen-binding fragment thereof. The pharmaceutical composition comprises 50 mg / mL to 60 mg / mL of sucrose, 50 mM to 60 mM of arginine or a pharma- ceutically acceptable salt thereof, 200 mM to 350 mM or 250 mM to 350 mM of proline, and / or 10 mM to 30 mM or 10 mM to 20 mM of sodium chloride, and the pH of the pharmaceutical composition is 4 to 7, 5 to 7, 5 to 6.5, 5.3 to 6.5, 5.3 to 6.3, 5.3 to 5.8, or 5.8 to 6.3.

[0036] In some embodiments, the pharmaceutical composition comprises (a) 30 mg / mL to 300 mg / mL of an anti-IL4Rα antibody or antigen-binding fragment thereof, (b) 1 mM to 100 mM of a phosphate buffer or a histidine salt buffer, (c) 0.01 mg / mL to 2 mg / mL of polysorbate 80, and (d) 60 mM to 600 mM of proline and 1 mM to 80 mM of sodium chloride, and optionally further comprises 10 mg / mL to 100 mg / mL of sucrose, and the pH of the pharmaceutical composition is 4 to 7, 5 to 7, 5 to 6.5, 5.3 to 6.5, 5.3 to 6.3, 5.3 to 5.8, or 5.8 to 6.3.

[0037] In some embodiments, the pharmaceutical composition comprises (a) 50 mg / mL to 250 mg / mL of an anti-IL4Rα antibody or antigen-binding fragment thereof, (b) 2 mM to 80 mM of a phosphate buffer or a histidine salt buffer, (c) 0.05 mg / mL to 1 mg / mL of polysorbate 80, and (d) 80 mM to 500 mM of proline and 2 mM to 60 mM of sodium chloride, and optionally further comprises 20 mg / mL to 80 mg / mL of sucrose, and the pH of the pharmaceutical composition is 4 to 7, 5 to 7, 5 to 6.5, 5.3 to 6.5, 5.3 to 6.3, 5.3 to 5.8, or 5.8 to 6.3.

[0038] In some embodiments, the pharmaceutical composition comprises (a) 70 mg / mL to 200 mg / mL of an anti-IL4Rα antibody or antigen-binding fragment thereof, (b) 4 mM to 60 mM of a phosphate buffer or a histidine salt buffer, (c) 0.1 mg / mL to 0.8 mg / mL of polysorbate 80, and (d) 100 mM to 450 mM or 120 mM to 400 mM of proline and 4 mM to 50 mM of sodium chloride, and optionally further comprises 30 mg / mL to 60 mg / mL of sucrose, and the pH of the pharmaceutical composition is 4 to 7, 5 to 7, 5 to 6.5, 5.3 to 6.5, 5.3 to 6.3, 5.3 to 5.8, or 5.8 to 6.3.

[0039] In some embodiments, the pharmaceutical composition comprises (a) 100 mg / mL to 180 mg / mL or 120 mg / mL to 180 mg / mL of an anti-IL4Rα antibody or antigen-binding fragment thereof, (b) 8 mM to 40 mM or 10 mM to 30 mM of a phosphate buffer or a histidine salt buffer, (c) 0.2 mg / mL to 0.6 mg / mL of polysorbate 80, and (d) 150 mM to 350 mM of proline and 8 mM to 40 mM of sodium chloride, and optionally further comprises 40 mg / mL to 50 mg / mL of sucrose, and the pH of the pharmaceutical composition is 4 to 7, 5 to 7, 5 to 6.5, 5.3 to 6.5, 5.3 to 6.3, 5.3 to 5.8, or 5.8 to 6.3.

[0040] In some embodiments, the pharmaceutical composition comprises (a) 120 mg / mL to 150 mg / mL or 150 mg / mL to 180 mg / mL of an anti-IL4Rα antibody or antigen-binding fragment thereof, (b) 10 mM to 20 mM or 20 mM to 30 mM of a phosphate buffer or a histidine salt buffer, (c) 0.2 mg / mL to 0.4 mg / mL of polysorbate 80, and (d) 200 mM to 350 mM or 250 mM to 350 mM of proline and 10 mM to 30 mM or 10 mM to 20 mM of sodium chloride, and optionally further comprises 50 mg / mL to 60 mg / mL of sucrose, and the pH of the pharmaceutical composition is 4 to 7, 5 to 7, 5 to 6.5, 5.3 to 6.5, 5.3 to 6.3, 5.3 to 5.8, or 5.8 to 6.3.

[0041] In some embodiments, the pharmaceutical composition comprises (a) 150 mg / mL of an anti-IL4Rα antibody or antigen-binding fragment thereof, (b) 20 mM histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL polysorbate 80, and (d) 50 mg / mL sucrose and 50 mM arginine hydrochloride, and the pH of the pharmaceutical composition is 5.8.

[0042] In some embodiments, the pharmaceutical composition comprises (a) 150 mg / mL of an anti-IL4Rα antibody or antigen-binding fragment thereof, (b) 20 mM histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL polysorbate 80, and (d) 250 mM proline and 20 mM sodium chloride, and the pH of the pharmaceutical composition is 5.8.

[0043] In some embodiments, the pharmaceutical composition comprises (a) 150 mg / mL of an anti-IL4Rα antibody or antigen-binding fragment thereof, (b) 20 mM histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL polysorbate 80, and (d) 350 mM proline, and the pH of the pharmaceutical composition is 5.8.

[0044] In some embodiments, the pharmaceutical composition comprises (a) 150 mg / mL of an anti-IL4Rα antibody or antigen-binding fragment thereof, (b) 20 mM histidine-histidine hydrochloride buffer, (c) 0.4 mg / mL polysorbate 80, and (d) 50 mg / mL sucrose, 250 mM proline, and 20 mM sodium chloride, and the pH of the pharmaceutical composition is 5.8.

[0045] In any embodiment, the anti-IL4Rα antibody or antigen-binding fragment thereof in the pharmaceutical composition comprises a heavy chain variable region, the heavy chain variable region comprising heavy chain variable region CDR1, heavy chain variable region CDR2 and heavy chain variable region CDR3 comprising amino acid sequences having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to the amino acid sequences represented by SEQ ID NOs: 1, 2 and 3, respectively.

[0046] In any embodiment, the anti-IL4Rα antibody or antigen-binding fragment thereof in the pharmaceutical composition comprises a heavy chain variable region, wherein the heavy chain variable region comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to the amino acid sequence represented by SEQ ID NO: 7, 8 or 9. Here, the amino acid sequence of SEQ ID NO: 8 is EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYGMSWVRQAPGKGLVX1VX2TINSNGGSTSYPDSVKGRFTISRDNAKNTLYLQMNSLRAEDTAVYYCARFFRFRNAMDYWGQGTLVTVSS (X1=W and X2=S, or X1=L and X2=A, or X1=W and X2=A).

[0047] In any embodiment, the anti-IL4Rα antibody or antigen-binding fragment thereof in the pharmaceutical composition comprises a light chain variable region, the light chain variable region comprising light chain variable region CDR1, light chain variable region CDR2 and light chain variable region CDR3 comprising amino acid sequences having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to the amino acid sequences represented by SEQ ID NOs: 4, 5 and 6, respectively.

[0048] In any embodiment, the anti-IL4Rα antibody or antigen-binding fragment thereof in the pharmaceutical composition comprises a light chain variable region, wherein the light chain variable region comprises an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to the amino acid sequence represented by SEQ ID NO: 10, 11 or 12. Here, the amino acid sequence of SEQ ID NO: 11 is DIQMTQSPSSLSASVGDRVTITCRTSENIYSYLAWYQQKPGKAPKX1LX2YNAKTLAEGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQHYYGPPTWTFGQGTKVEIK (X1=L and X2=I, or X1=F and X2=V, or X1=F and X2=I).

[0049] In any embodiment, the anti-IL4Rα antibody or antigen-binding fragment thereof in the pharmaceutical composition comprises a heavy chain variable region and a light chain variable region, each of which comprises CDR1, CDR2 and CDR3, wherein the heavy chain variable region CDR1, the heavy chain variable region CDR2, the heavy chain variable region CDR3, the light chain variable region CDR1, the light chain variable region CDR2 and the light chain variable region CDR3 comprise amino acid sequences that have at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to the amino acid sequences represented by SEQ ID NOs: 1, 2, 3, 4, 5 and 6, respectively.

[0050] In any embodiment, the anti-IL4Rα antibody or antigen-binding fragment thereof in the pharmaceutical composition comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region and the light chain variable region comprising: (1) an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to the amino acid sequences represented by SEQ ID NOs: 7 and 10, respectively; (2) an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to the amino acid sequences represented by SEQ ID NOs: 8 and 11, respectively (X1=W and X2=S in SEQ ID NO: 8); (3) an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to the amino acid sequences represented by SEQ ID NOs: 8 and 12, respectively (X1=W and X2=S in SEQ ID NO: 8); (4) an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to the amino acid sequences represented by SEQ ID NOs: 9 and 11, respectively; (5) an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to the amino acid sequences represented by SEQ ID NOs: 9 and 12, respectively; (6) an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to the amino acid sequences represented by SEQ ID NOs: 8 and 11, respectively (X1=L and X2=A in SEQ ID NO: 8); (7) an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to the amino acid sequences represented by SEQ ID NOs: 8 and 12, respectively (X1=L and X2=A in SEQ ID NO: 8); (8) an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to the amino acid sequences represented by SEQ ID NOs: 8 and 11, respectively (X1=W and X2=A in SEQ ID NO: 8); or (9) An amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to the amino acid sequences represented by SEQ ID NOs: 8 and 12, respectively (X1=W and X2=A in SEQ ID NO: 8). wherein the amino acid sequence of SEQ ID NO:8 is EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYGMSWVRQAPGKGLVX1VX2TINSNGGSTSYPDSVKGRFTISRDNAKNTLYLQMNSLRAEDTAVYYCARFFRFRNAMDYWGQGTLVTVSS, The amino acid sequence of SEQ ID NO:11 is DIQMTQSPSSLSASVGDRVTITCRTSENIYSYLAWYQQKPGKAPKX1LX2YNAKTLAEGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQHYYGPPTWTFGQGTKVEIK (X1=L and X2=I, X1=F and X2=V, or X1=F and X2=I).

[0051] In any embodiment, the anti-IL4Rα antibody or antigen-binding fragment thereof in the pharmaceutical composition comprises a heavy chain comprising a heavy chain variable region and a heavy chain constant region, and a light chain comprising a light chain variable region and a light chain constant region, wherein the C-terminus of the heavy chain variable region is linked to the N-terminus of the heavy chain constant region, and the C-terminus of the light chain variable region is linked to the N-terminus of the light chain constant region, wherein the heavy chain variable region and the light chain variable region comprise the amino acid sequences described above, the heavy chain constant region has a human IgG4 constant region having the amino acid sequence shown in SEQ ID NO:13, and the light chain constant region has a human κ constant region having the amino acid sequence shown in SEQ ID NO:14.

[0052] In any embodiment, the anti-IL4Rα antibody or antigen-binding fragment thereof in the pharmaceutical composition may be a full-length antibody, for example, a full-length antibody of IgG1, IgG2 or IgG4 isotype, preferably a full-length antibody of IgG4 isotype. In other embodiments, the anti-IL4Rα antibody or antigen-binding fragment thereof in the pharmaceutical composition may be a single-chain variable domain (scFv) antibody or an antibody fragment such as a Fab or F(ab')2 fragment.

[0053] In any embodiment, the anti-IL4Rα antibody or antigen-binding fragment thereof in the pharmaceutical composition is the C2C1A1A1 antibody described in patent application no. PCT / CN2021077784.

[0054] The present disclosure further provides a method for producing the pharmaceutical composition, comprising the step of contacting the anti-IL4Rα antibody or antigen-binding fragment thereof with a buffer. The step includes, for example, replacing the anti-IL4Rα antibody or antigen-binding fragment thereof with a buffer. The buffer includes a phosphate buffer, an acetate buffer, a histidine buffer, and the like. The production method further includes adding a surfactant and a stabilizer in any order. The stabilizer includes trehalose, mannitol, sucrose, arginine or a pharma- ceutically acceptable salt thereof, proline, and / or sodium chloride, and the surfactant includes a polysorbate, a poloxamer, a polyethylene glycol, or a polyhydroxy substance.

[0055] The present disclosure further provides a method for producing a lyophilized formulation comprising an anti-IL4Rα antibody or an antigen-binding fragment thereof, comprising the step of lyophilizing the pharmaceutical composition. In some embodiments, the lyophilization is performed according to methods known in the art, including, but not limited to, the steps of pre-freezing, primary drying, and secondary drying. It will be understood by those skilled in the art that any method for removing water from the pharmaceutical composition of the present disclosure is applicable to the present disclosure.

[0056] The present disclosure further provides a lyophilized preparation comprising an anti-IL4Rα antibody or an antigen-binding fragment thereof, produced by the above-mentioned method for producing a lyophilized preparation.

[0057] The present disclosure further provides a lyophilized formulation comprising an anti-IL4Rα antibody or antigen-binding fragment thereof, which can be dissolved to form the above-described pharmaceutical composition.

[0058] The present disclosure further provides an article of manufacture comprising a container filled with the pharmaceutical composition or the lyophilized formulation.

[0059] The pharmaceutical composition or lyophilized formulation of the present disclosure can be administered by known methods. For example, it can be injected or infused over a period of time by a suitable method, for example, subcutaneously, intravenously, intraperitoneally, intramuscularly, intraarterially, intralesionally or intraarticularly, locally, by inhalation, or by sustained or delayed release. The pharmaceutical composition or lyophilized formulation of the present disclosure can be diluted to an appropriate concentration with a suitable diluent before administration to provide an optimal desired response (e.g., therapeutic response).

[0060] The present disclosure provides a method for reducing IL4 and / or IL13 signaling in a subject, comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition, lyophilized formulation or product of the present disclosure. IL4 signals through receptors including IL-4Rα and γc, and IL13 signals through receptors including IL-4Rα and IL13Rα1. IL4 and / or IL13 signaling is manifested as activation and / or proliferation of B cells, eosinophils, macrophages, proliferation of fibroblasts, and proliferation of smooth muscle cells (such as proliferation of airway smooth muscle cells).

[0061] The present disclosure provides a method of reducing a type 2 immune response in a subject comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition, lyophilized formulation, or product of the disclosure.

[0062] The present disclosure provides a method for treating a disease associated with excessive IL4 and / or IL13 signaling in a subject, comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition, lyophilized formulation or product of the present disclosure, wherein the disease associated with excessive IL4 and / or IL13 signaling is an allergic disease, non-limiting examples of which include atopic dermatitis, allergic reactions, allergic rhinitis or allergic asthma.

[0063] The present disclosure provides a method for treating a tumor associated with increased STAT6 activation in a subject, comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition, lyophilized formulation, or product of the present disclosure. In some embodiments, the tumor comprises a solid tumor or a non-solid tumor. In some embodiments, examples of the tumor include, but are not limited to, melanoma, lung cancer, renal cancer, prostate cancer, cervical cancer, colorectal cancer, gastric cancer, pancreatic cancer, ovarian cancer, and urothelial cancer.

[0064] Other features and advantages of the present disclosure will become more apparent from the following detailed description and examples, which are not intended to be limiting. The contents of all references, Genbank records, patents and published patent applications cited in this application are expressly incorporated herein by reference. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0065] To facilitate understanding of this disclosure, some terms are first defined. Other definitions are provided in the detailed description. Unless otherwise defined herein, all other technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0066] "Buffer" refers to a pharma- ceutically acceptable substance or mixture of substances that can maintain the pH of a pharmaceutical composition in a desired pH range. Examples of buffers that can be applied to the pharmaceutical composition of the present disclosure include phosphate buffers, acetate buffers, or histidine buffers.

[0067] A "phosphate buffer" is a buffer containing phosphate ions. For example, it includes a buffer containing phosphoric acid and / or a phosphate salt, where phosphoric acid includes phosphoric acid and / or its hydrate, and phosphate salt includes phosphate salt and / or its hydrate. Examples of phosphate buffer include, but are not limited to, sodium phosphate buffer or potassium phosphate buffer, and a preferred phosphate buffer is sodium phosphate buffer. In one particular example, the sodium phosphate buffer includes sodium dihydrogen phosphate monohydrate (NaH2PO4·H2O) and disodium hydrogen phosphate dodecahydrate (Na2HPO4·12H2O).

[0068] An "acetate buffer" is a buffer containing acetate ions. For example, it includes a buffer containing acetic acid and / or acetate, where acetic acid includes acetic acid and / or its hydrates, and acetate includes acetate and / or its hydrates. Examples of acetate buffers include, but are not limited to, potassium acetate buffer, ammonium acetate buffer, or sodium acetate buffer, and sodium acetate-acetic acid buffer, and a preferred acetate buffer is sodium acetate-acetic acid buffer. Sodium acetate-acetic acid buffer contains acetic acid and sodium acetate, where acetic acid includes acetic acid and / or its hydrates, and sodium acetate includes sodium acetate and / or its hydrates. In a particular example, sodium acetate-acetic acid buffer contains acetic acid and sodium acetate trihydrate (CH3COONa·3H2O).

[0069] A "histidine salt buffer" is a buffer containing histidine ions. For example, it includes a buffer containing histidine and / or a histidine salt, where histidine includes histidine and / or its hydrate, and histidine salt includes histidine salt and / or its hydrate. Examples of histidine salt buffers include histidine-hydrochloric acid buffer, histidine-acetate buffer, histidine-histidine hydrochloride, or histidine hydrochloride buffer, and the preferred histidine salt buffer is histidine-histidine hydrochloride buffer. Histidine-histidine hydrochloride includes histidine and histidine hydrochloride, where histidine includes histidine and / or its hydrate, and histidine hydrochloride includes histidine hydrochloride and / or its hydrate. In one particular example, the histidine-histidine hydrochloride buffer comprises histidine and histidine hydrochloride monohydrate (C6H9N3O2·HCl·H2O).

[0070] "Stabilizer" refers to a pharma- ceutically acceptable substance or mixture of substances for maintaining the stability of an active ingredient in a pharmaceutical composition. In the present disclosure, the stabilizer also functions as a viscosity-lowering agent and / or an isotonicity agent, etc.

[0071] "Pharmaceutical composition" refers to any combination of specific active ingredients (e.g., antibodies) in a specific amount, and any combination of specific active ingredients in a specific amount, either directly or indirectly. The purpose of a pharmaceutical composition is to make the active ingredients suitable for manufacture and administration to a patient, and to maintain biological activity and / or stability during storage and subsequent use. In some embodiments, the pharmaceutical composition is an aqueous soluble injectable formulation. The aqueous soluble injectable formulation includes a water-soluble formulation that is not lyophilized or that is reconstituted from a lyophilized powder. In another embodiment, the pharmaceutical composition is a lyophilized formulation. In this disclosure, "pharmaceutical composition" and "formulation" are not mutually exclusive.

[0072] A "stable" or "stabilized" pharmaceutical composition refers to a pharmaceutical composition in which an active ingredient (e.g., an antibody) substantially retains its physical stability and / or chemical stability and / or biological activity when stored therein. Various analytical techniques for measuring the stability of an active ingredient are known in the art and are described, for example, in Peptide and Protein Drug Delivery, 247-301, Vincent Lee Ed., Marcel Dekker, Inc., New York, NY, Pubs. (1991) Jones, A. Adv. Drug Delivery Rev. 10:29-90 (1993). Stability can be measured at a selected temperature, for a selected period of time, and other storage conditions. For example, an active ingredient is considered to "retain its physical stability" in a pharmaceutical composition if the active ingredient does not show significant increases in aggregation, precipitation, and / or denaturation as determined by visual inspection of color and / or clarity, or by UV light scattering, size exclusion chromatography (SEC), differential scanning calorimetry (DSC), or differential scanning fluorescence (DSF). Preferably, when using the pharmaceutical composition of the present disclosure, no more than 10%, no more than 5%, no more than 4% of the active ingredient forms aggregates (also called polymeric impurities) as measured by SEC or other suitable methods used to measure aggregate formation. An active ingredient (e.g., an antibody) is considered to "retain its chemical stability" in the pharmaceutical composition if the active ingredient does not show significant chemical changes. Chemical stability can be evaluated by detection and quantification of chemically altered antibodies. Processes that generally alter the chemical structure of proteins include hydrolysis and cleavage (assessed by methods such as size exclusion chromatography and SDS-PAGE), oxidation (assessed by methods such as mass spectrometry and peptide mapping in combination with MALDI / TOF / MS), deamidation (assessed by methods such as ion exchange chromatography, capillary isoelectric focusing, peptide mapping, isoaspartic acid measurement), and isomerization (assessed by measuring isoaspartic acid content, peptide mapping, etc.).An active ingredient is considered to "retain its biological activity" in a pharmaceutical composition for a given period of time if the biological activity of the active ingredient (e.g., an antibody) for that period of time is within a given range of biological activity indicated at the time of manufacture of the pharmaceutical composition, which can be confirmed, for example, by an antigen binding assay.

[0073] "Macromolecular weight impurities" or "aggregates" are a collective term for impurities that have a larger molecular weight than the active ingredient of interest (e.g., an antibody).

[0074] "Charge variant" refers to a variant in which the charge of an antibody molecule is changed directly or indirectly by glycosylation, deamidation, oxidation, and / or isomerization of the antibody. These charge variants can be detected by capillary isoelectric focusing (CIEF) or cation exchange chromatography (CEX-HPLC), etc.

[0075] The articles "a," "an," and "the" are used to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. For example, "a pharmaceutical composition" may refer to one pharmaceutical composition or to multiple pharmaceutical compositions.

[0076] "About" or "approximately" means that the value is within an acceptable error range for a particular value as determined by one of ordinary skill in the art, depending in part on how the value is measured or determined (i.e., the limitations of the measurement system). For example, in the art, "about" or "approximately" can mean within 1 or more than 1 standard deviation. Alternatively, "about" or "approximately" can mean a range of plus or minus 15%, 10%, 5% or 1% for such value. Furthermore, particularly for biological systems or processes, the term can mean within an order of magnitude of a value or within 5 times the value. Unless otherwise specified, "about XX" or "approximately XX" or "substantially including XX" means a value within an acceptable error range for a particular value "XX" (including the value "XX" itself and values ​​within an acceptable error range for this value as determined by one of ordinary skill in the art).

[0077] As used herein, any percentage range, ratio range, or integer range, unless otherwise specified, is understood to include any integer value within the recited range and, where appropriate, fractions thereof (such as 1 / 10 and 1 / 100 of an integer).

[0078] In this specification, unless the context indicates otherwise, the terms "comprise", "include", and "contain" mean including the recited step or element or group of steps or elements, but not excluding any other step or element or group of steps or elements. "Consisting of" means including and limited to the elements that follow "consisting of". Thus, the phrase "consisting of" indicates that the recited elements are necessary or essential, and no other elements are present. "Consisting essentially of" means including any elements recited before this phrase, and limited to other elements that do not interfere with or contribute to the activity or action described in the present invention with respect to those recited elements. Thus, the phrase "consisting essentially of" means that the recited elements are necessary or essential, but other elements are optional and may or may not be present depending on whether they affect the activity or action of the recited elements.

[0079] The term "IL4Rα" refers to the interleukin 4 receptor α subunit. The term "IL4Rα" includes variants, isoforms, homologs, orthologs, and paralogs. For example, in some cases, an antibody specific for human IL4Rα protein may cross-react with IL4Rα protein of a species other than human (e.g., monkey). In other embodiments, an antibody specific for human IL4Rα protein has no cross-reactivity with other species or types of proteins, is completely specific for human IL4Rα protein, or is cross-reactive with IL4Rα of only a particular species and not all other species.

[0080] The term "human IL4Rα" refers to an IL4Rα protein having a human amino acid sequence, such as the amino acid sequence of human IL4Rα in Genbank Accession No. NP_001244335.1. The terms "cynomolgus IL4Rα" and "marmoset IL4Rα" refer to IL4Rα sequences having, for example, the amino acid sequences of Genbank Accession Nos. EHH60265.1 and NP_001244161.1, respectively.

[0081] The term "antibody" as used herein includes full length antibodies and any antigen-binding fragments thereof or single chains thereof. Full length antibodies are glycoproteins that contain two heavy (H) chains and two light (L) chains, which are linked via disulfide bonds. Each of the heavy chains contains a heavy chain variable region (V H ) and the heavy chain constant region. The heavy chain constant region is H1 , C H2 and C H3 Each light chain is composed of three domains: L ) and the light chain constant region. The light chain constant region consists of one domain, C L It consists of: V H and V L The region can be divided into hypervariable regions, also called complementarity determining regions (CDRs), interspersed with more conserved framework regions (FRs). H and V L consists of 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 antigens. The constant region of the antibody can mediate the binding of the immunoglobulin to host tissues or factors, including various immune system cells (e.g., effector cells) and the first component (C1q) of the classical complement system.

[0082] As used herein, an "antigen-binding fragment" of an antibody (or abbreviated as "antibody fragment") refers to one or more fragments of an antibody that retain the ability to specifically bind to an antigen (e.g., IL4Rα protein). It has been revealed that the antigen-binding function of an antibody can be exerted by a fragment of a full-length antibody. Examples of antibody fragments included in the "antigen-binding fragment" of an antibody include (i) V L , V H , C L and C H1 (ii) a F(ab')2 fragment, which is a bivalent fragment comprising two Fab fragments linked by disulfide bonds in the hinge region; and (iii) a V H and C. H1 (iv) a single-arm V of an antibody L and V H (v) an Fv fragment consisting of V H (vi) isolated complementarity determining regions (CDRs); and (vii) a heavy chain variable region comprising one variable domain and two constant domains. In addition, the two domains of the Fv fragment, V, are included. L and V H are encoded by separate genes, which can be recombined to produce L and V H can be linked by a synthetic linker that allows them to be produced as a single protein chain that pairs to form a monovalent molecule (known as a single-chain Fv (scFv); see, e.g., Bird et al., (1988) Science 242:423-426 and Huston et al., (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883). Such single-chain antibodies are also encompassed by the term "antigen-binding fragment" of an antibody. These antibody fragments can be obtained by conventional techniques known to those of skill in the art, and can be screened for utility in the same manner as full-length antibodies.

[0083] As used herein, the term "isolated antibody" refers to an antibody that is substantially free of other antibodies having different antigenic specificities (e.g., an isolated antibody that specifically binds to an IL4Rα protein is substantially free of antibodies that specifically bind to antigens other than the IL4Rα protein). However, an isolated antibody that specifically binds to a human IL4Rα protein may have cross-reactivity to other antigens, such as, for example, IL4Rα proteins from other species. Furthermore, an isolated antibody is substantially free of other cellular material and / or chemicals.

[0084] As used herein, the term "murine antibody" refers to an antibody in which both the framework and CDR regions of the variable regions are derived from mouse germline immunoglobulin sequences. Furthermore, if the antibody contains a constant region, the constant region is also derived from mouse germline immunoglobulin sequences. The murine antibodies herein may include amino acid residues not encoded by mouse germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo). However, the term "murine antibody" as used herein does not include antibodies in which CDR sequences from another mammalian species have been grafted onto murine framework sequences.

[0085] The term "chimeric antibody" refers to an antibody that combines genetic material of non-human origin with genetic material of human origin, or more broadly, a chimeric antibody refers to an antibody that has genetic material from one species and genetic material from another species.

[0086] As used herein, the term "humanized antibody" refers to an antibody derived from a non-human species but whose protein sequence has been altered to increase similarity to natural human antibodies. In some specific examples, a "humanized antibody" contains complementarity determining regions (CDRs) from a non-human antibody and framework regions (FRs) and constant regions from a human antibody.

[0087] The term "isotype" refers to the antibody class (e.g., IgM or IgG1) that is encoded by heavy chain constant region genes.

[0088] The phrases "antibody that recognizes an antigen" and "antibody specific for an antigen / antibody having specificity for an antigen" are used interchangeably herein with the term "antibody that specifically binds to an antigen."

[0089] As used herein, an antibody that "specifically binds to human IL4Rα" means an antibody that binds to human IL4Rα protein (and optionally IL4Rα proteins of one or more non-human animal species), but does not substantially bind to non-IL4Rα proteins. Preferably, the antibody binds to human IL4Rα protein with "high affinity," i.e., greater than 5.0x. -8 M or less, preferably 1.0×10 -8 M or less, preferably 7.0×10 -9 K below M D It binds to the human IL4Rα protein.

[0090] As used herein, "does not substantially bind" to a protein or cell means that it does not bind to a protein or cell or does not bind with high affinity, i.e., less than 1.0×10 -6 M or more, preferably 1.0×10 -5 M or more, preferably 1.0×10 -4 M or more, preferably 1.0×10 -3 M or more, preferably 1.0×10 -2 K over M D This means that the molecule binds to a protein or cell via the agonist.

[0091] The term "high affinity" refers to an IgG antibody that binds to an antigen with a binding affinity of 1.0 x 10 -6 M or less, preferably 5.0×10 -8 M or less, preferably 1.0×10 -8 M or less, preferably 7.0×10 -9 M or less, preferably 1.0×10 -9 K below M DHowever, for other antibody isotypes, the meaning of "high affinity" may be different. For example, for the IgM isotype, "high affinity" means that the antibody has a binding affinity of 1.0 x 10 -6 M or less, preferably 1.0×10 -7 M or less, preferably 1.0×10 -8 K below M D It means having

[0092] As used herein, the term "K assoc " or "K a " refers to the binding rate of a particular antibody-antigen interaction, and the term "K dis " or "K d " refers to the off-rate of a particular antibody-antigen interaction. D " is K a K for d The ratio (i.e. K d / K a ) and refers to the dissociation constant expressed as molar concentration (M). D The K value can be measured by methods well known in the art. D A preferred method for measuring the value is a method using surface plasmon resonance, preferably, for example, Biacore. TM The method uses a biosensor system such as a fluorosensor system.

[0093] The term “EC 50 ", also called the median effective concentration, refers to the antibody concentration that induces a response halfway between the baseline and maximum after a specific exposure time. 50 ", also referred to as half-maximal inhibitory concentration, refers to the concentration of an antibody that inhibits a specific biological or biochemical function by 50% compared to the absence of the antibody.

[0094] The term "subject" includes any human or non-human animal. The term "non-human animal" encompasses all vertebrates, such as mammals and non-mammals, including, for example, non-human primates, sheep, dogs, cats, cows, horses, chickens, amphibians, reptiles, but is preferably a mammal, including, for example, non-human primates, sheep, dogs, cats, cows, and horses.

[0095] The term "therapeutically effective amount" refers to an amount of an antibody of the present invention sufficient to result in prevention or amelioration of symptoms associated with a disease or disorder (e.g., cancer) and / or a reduction in the severity of the disease or disorder. The therapeutically effective amount is related to the disease being treated, and the actual effective amount can be easily determined by one of ordinary skill in the art.

[0096] The term "identity" refers to the sequence similarity between two polynucleotide sequences or two polypeptides. Sequence comparison and percent identity between two sequences can be determined by the default settings of the algorithms BLASTN / BLASTP on the website of the National Center for Biotechnology Information.

[0097] The terms "Xn" and "Xaa" are the same and refer to an unspecified amino acid, the scope of which is defined by the definition in the relevant description.

[0098] Each aspect of the invention is described in more detail below.

[0099] Anti-IL4Rα antibody or antigen-binding fragment thereof The anti-IL4Rα antibodies or antigen-binding fragments thereof of the present disclosure specifically bind to human IL4Rα and have comparable (if not superior) binding affinity / capacity compared to reported anti-IL4Rα antibodies (e.g., Dupilumab).

[0100] The anti-IL4Rα antibody or antigen-binding fragment thereof of the present disclosure can block the binding of IL4Rα to IL4 or IL13-IL13Rα1, thereby blocking intracellular signal transduction. It has equal or greater blocking activity compared to reported anti-IL4Rα antibodies (e.g., Dupilumab).

[0101] Preferably, the anti-IL4Rα antibodies of the invention are humanized monoclonal antibodies. Additionally, or alternatively, the anti-IL4Rα antibodies of the invention can be, for example, murine, chimeric or human monoclonal antibodies.

[0102] The anti-IL4Rα antibodies of the present disclosure are monoclonal antibodies that are structurally and chemically characterized as described below and in the Examples. The amino acid sequence numbers of the heavy / light chain variable regions of the antibodies are summarized in Table 1 below. Some antibodies have the same V H Or V L The heavy chain constant region of the antibody may be a human IgG4 heavy chain constant region having the amino acid sequence represented by SEQ ID NO:13, and the light chain constant region of the antibody may be a human κ constant region having the amino acid sequence represented by SEQ ID NO:14.

[0103] [Table 1]

[0104] The heavy chain variable region CDRs and light chain variable region CDRs in Table 1 are defined according to the Kabat numbering system. However, as known in the art, CDR regions can also be determined based on the heavy / light chain variable region sequences using other numbering systems, such as the Chothia, IMGT, AbM or Contact numbering systems / methods. When an antibody or antigen-binding fragment thereof is characterized by a specific CDR sequence, the scope of said antibody includes an antibody or antigen-binding fragment thereof characterized by a CDR sequence defined by any numbering system in the art (e.g., the Chothia, IMGT, AbM or Contact numbering system).

[0105] V of other anti-IL4Rα antibodies that bind to human IL4Rα H and V L The V sequence (or CDR sequence) of the anti-IL4Rα antibody of the present disclosure H and V L The sequences (or CDR sequences) can be "mixed and matched". H and V L When mixing and pairing the chains (or the CDRs in those chains), a particular V H / V L Pair of V H The sequence is structurally similar to V H Similarly, a particular V H / V L Pair of V L The sequence is also structurally similar to V L It is replaced by the sequence.

[0106] Thus, in one embodiment, the anti-IL4Rα antibody or antigen-binding fragment thereof of the disclosure: (a) a heavy chain variable region comprising an amino acid sequence shown in Table 1; (b) the V amino acid sequence shown in Table 1, or another anti-IL4Rα antibody (the antibody or antigen-binding fragment thereof specifically binds to human IL4Rα). L and a light chain variable region comprising:

[0107] In another embodiment, the anti-IL4Rα antibody or antigen-binding fragment thereof of the present disclosure comprises: (a) CDR1, CDR2 and CDR3 of a heavy chain variable region shown in Table 1; (b) CDR1, CDR2 and CDR3 of a light chain variable region shown in Table 1, or the CDRs of another anti-IL4Rα antibody (the antibody or antigen-binding fragment thereof specifically binds to human IL4Rα).

[0108] In another embodiment, an anti-IL4Rα antibody or antigen-binding fragment thereof of the present disclosure comprises CDR2 of the heavy chain variable region of an anti-IL4Rα antibody of the present disclosure and CDRs of another antibody that binds to human IL4Rα, e.g., CDR1 and / or CDR3 of the heavy chain variable region, and / or CDR1, CDR2 and / or CDR3 of the light chain variable region from another anti-IL4Rα antibody.

[0109] Furthermore, it is well known in the art that the binding specificity of an antibody for a homologous antigen can be determined by the CDR3 domain alone, independent of the CDR1 and / or CDR2 domains, and that multiple antibodies with the same binding specificity can be predictably generated based on a common CDR3 sequence.

[0110] Thus, in another embodiment, the anti-IL4Rα antibody or antigen-binding fragment thereof of the present disclosure comprises the CDR2 of the heavy chain variable region of the anti-IL4Rα antibody of the present disclosure and at least the CDR3 of the heavy chain variable region and / or the light chain variable region of the anti-IL4Rα antibody of the present disclosure, or the CDR3 of the heavy chain variable region and / or the light chain variable region of another anti-IL4Rα antibody that specifically binds to human IL4Rα. Preferably, these antibodies and the anti-IL4Rα antibody of the present disclosure (a) compete to bind to IL4Rα, (b) retain functional characteristics, (c) bind to the same epitope, and / or (d) have similar binding affinity. In another embodiment, the anti-IL4Rα antibody of the present disclosure may further comprise the CDR2 of the light chain variable region of the anti-IL4Rα antibody of the present disclosure, or the CDR2 of the light chain variable region of another anti-IL4Rα antibody that specifically binds to human IL4Rα. In another embodiment, the anti-IL4Rα antibody of the present disclosure may further comprise CDR1 of the heavy chain variable region and / or light chain variable region of the anti-IL4Rα antibody of the present disclosure, or CDR1 of the heavy chain variable region and / or light chain variable region of another anti-IL4Rα antibody that specifically binds to human IL4Rα.

[0111] In another embodiment, an anti-IL4Rα antibody or antigen-binding fragment thereof of the present disclosure comprises a heavy chain variable region and / or a light chain variable region that comprises a CDR1, CDR2, and CDR3, respectively, that differ from the sequences of the CDR1, CDR2, and CDR3 of an anti-IL4Rα antibody of the present disclosure by one or more conservative modifications. It is understood in the art that certain conservative sequence modifications that do not eliminate antigen binding can be made. See, e.g., Brummell et al., (1993) Biochem 32:1180-8; de Wildt et al., (1997) Prot. Eng. 10:835-41; Komissarov et al., (1997) J. Biol. Chem. 272:26864-26870; Hall et al., (1992) J. Immunol. 149:1605-12; Kelley and O'Connell (1993) Biochem. 32:6862-35; Adib-Conquy et al., (1998) Int. Immunol. 10:341-6 and Beers et al., (2000) Clin. Can. Res. 6:2835-43.

[0112] Thus, in one embodiment, an anti-IL4Rα antibody or antigen-binding fragment thereof of the present disclosure comprises a heavy chain variable region and / or a light chain variable region comprising CDR1, CDR2, and CDR3, respectively, wherein: (a) the CDR1 of the heavy chain variable region comprises a sequence shown in Table 1, and / or a conservative modification thereof; and / or (b) the CDR2 of the heavy chain variable region comprises a sequence set forth in Table 1, and / or a conservative modification thereof; and / or (c) the CDR3 of the heavy chain variable region comprises a sequence set forth in Table 1, and / or a conservative modification thereof; and / or (d) CDR1, and / or CDR2, and / or CDR3 of the light chain variable region comprise a sequence set forth in Table 1, and / or a conservative modification thereof; and (e) the antibody or antigen-binding fragment thereof specifically binds to human IL4Rα.

[0113] As used herein, the term "conservative sequence modifications" refers to amino acid modifications that do not specifically affect or change the binding properties of an antibody. Such conservative modifications include amino acid substitutions, insertions, and deletions. Modifications can be introduced into the antibodies of the present disclosure by standard techniques known in the art, such as, for example, site-directed mutagenesis and PCR-mediated mutagenesis. A conservative amino acid substitution refers to the replacement of an amino acid residue with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains are known in the art. These families of amino acid residues include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), β-branched side chains (e.g., threonine, valine, isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). In this manner, one or more amino acid residues in the CDR regions of the anti-IL4Rα antibodies of the present disclosure can be replaced with other amino acid residues from the same side chain family, and the resulting antibodies can be tested for retained function (i.e., the functions described above) by the functional testing methods described herein.

[0114] The pharmaceutical compositions, lyophilized formulations or products of the disclosure have numerous in vitro and in vivo utilities, including, for example, the treatment of diseases associated with excessive IL4 and / or IL13 signaling.

[0115] In one aspect, the anti-IL4Rα antibody or antigen-binding fragment thereof can reduce type 2 immunity by blocking the binding of IL4Rα to IL4 or IL13-IL13Rα1. The present disclosure provides a method of treating a type 2 immunity-related allergic disease in a subject, comprising administering to the subject a pharmaceutical composition, lyophilized formulation, or product of the present disclosure. The allergic disease can be atopic dermatitis, allergic reaction, allergic rhinitis, or allergic asthma.

[0116] In another aspect, IL4 or IL13 signaling can activate STAT6, and STAT6 inhibitors have been found to inhibit the proliferation of cancer cells. The present disclosure provides a method for inhibiting the proliferation of tumor cells in a subject, comprising administering to the subject a pharmaceutical composition, lyophilized formulation or product of the present disclosure. Non-limiting examples of the tumor include, but are not limited to, melanoma, lung cancer, kidney cancer, prostate cancer, cervical cancer, colorectal cancer, gastric cancer, pancreatic cancer, ovarian cancer and urothelial cancer.

[0117] In another aspect, the present disclosure provides a method of reducing or inhibiting activation of cells responsive to IL-4 or IL-13 in a subject. In some embodiments, inhibiting activation comprises inhibiting cytokine production or secretion. In some embodiments, inhibiting activation comprises inhibiting proliferation. Cells that respond to IL-4 by activating a hybrid IL-4Rα / γc receptor include, but are not limited to, B cells, eosinophils, and macrophages. Cells that respond to IL-13 by activating a hybrid IL-4Rα / IL-13Rα1 receptor include, but are not limited to, fibroblasts and smooth muscle cells. Thus, in one embodiment, the present disclosure provides a method of inhibiting proliferation of smooth muscle cells. In another embodiment, the present disclosure provides a method of inhibiting proliferation of fibroblasts.

[0118] The present invention is further illustrated by the following examples, which are not intended to be limiting. All figures in this application and all references, Genebank sequences, patents and published patent applications cited in this application are expressly incorporated herein by reference. EXAMPLES

[0119] Methods for preparing and purifying the anti-IL4Rα antibodies of the present disclosure are described in patent application PCT / CN2021077784, the entire contents of which are incorporated herein by reference.

[0120] (Example 1) Affinity assay of anti-IL4Rα antibodies by BIACORE surface plasmon resonance The binding affinity and binding kinetics of the anti-IL4Rα antibodies (mAbs) of the present disclosure were measured using a Biacore T200 system (GE Healthcare, Pittsburgh, PA, USA).

[0121] Briefly, goat anti-human IgG (GE healthcare, Catalog BR100839, Human Antibody Capture Kit) was covalently coupled to a CM5 chip (carboxymethylated dextran coated chip) via primary amine groups using a standard amine coupling kit provided with Biacore (GE Healthcare, Pittsburgh, PA, USA). Unreacted sites on the biosensor surface were blocked with ethanolamine. Next, the anti-IL4Rα antibody of the present disclosure at a concentration of 66.67 nM and the anti-IL4Rα antibody reference substance (Dupilumab®, also known as BM) at a concentration of 10 μg / mL were flowed through the chip at a flow rate of 10 μL / min. Then, recombinant human IL4Rα-his protein (prepared in-house, amino acid sequence as shown in SEQ ID NO: 15), cynomolgus monkey IL4Rα-his protein (Sino biological inc., catalog 90897-C08H), or marmoset IL4Rα-his protein (custom-made by Sino biological inc., also called cal-IL4Rα-his, amino acid sequence as shown in SEQ ID NO: 16) serially diluted with HBSEP buffer (from Biacore) were flowed through the chip at a flow rate of 30 μL / min. The antigen-antibody binding rate was tracked for 2 minutes, and the dissociation rate was tracked for 10 minutes. The binding and dissociation curves were fitted to a 1:1 Langmuir binding model using the software BIA evaluation, and the K D , K a and K d The value was calculated.

[0122] (Example 2) Binding activity of anti-IL4Rα antibody to IL4Rα The binding activity of the anti-IL4Rα antibodies of the present disclosure to IL4Rα was measured by capture ELISA, flow cytometry (FACS), and indirect ELISA.

[0123] 2.1 Capture ELISA Briefly, affinity purified goat anti-human IgG (Fcγ fragment specificity, Jackson Immunoresearch, Cat. 109-005-098) dissolved in PBS at a concentration of 2 μg / mL was coated at 100 μL / well onto 96-well plates and incubated overnight at 4° C. Plates were washed once with wash buffer (PBS+0.05% w / v Tween-20, PBST) before adding blocking buffer (5% w / v skim milk in PBST) at 200 μL / well and blocking for 2 h at 37° C. The plate was washed again, and 100 μL of serially diluted (5-fold serial dilution from a starting concentration of 66.7 nM in PBST containing 2.5% w / v skim milk) anti-IL4Rα antibodies of the present disclosure, reference substances, or negative control hIgG (human immunoglobulin for intravenous injection (pH 4), Hualan Biological Engineering Inc.) was added to each well, and co-incubated at 37° C. for 40 minutes, after which the plate was washed four times. Biotin-labeled human IL4Rα-his protein (prepared in-house, amino acid sequence as shown in SEQ ID NO: 15, dissolved at a concentration of 0.14 nM in PBST containing 2.5% w / v skim milk) was added at 100 μL / well to the 96-well plate on which the anti-IL4Rα antibodies were captured, and co-incubated at 37° C. for 40 minutes, after which the plate was washed four times. Then, 100 μL / well of HRP-conjugated streptavidin (diluted 1:10000 in PBST, Jackson Immunoresearch, catalog 016-030-084) was added and incubated for 40 min at 37°C. After a final wash, 100 μL / well of ELISA substrate TMB (Innoreagents, catalog TMB-S-002) was added and incubated. After 10 min at 25°C, the reaction was stopped by adding 50 μL / well of 1M H2SO4 and the absorbance at 450 nm was measured. Data analysis was performed with the software Graphpad Prism, and EC 50 The value was calculated.

[0124] 2.2 Cell-based FACS binding The binding activity of the anti-IL4Rα antibodies of the present disclosure to IL4Rα expressed on the surface of 293F-IL4Rα cells was analyzed by flow cytometry (FACS). Briefly, 293F cells (Thermofisher Inc., catalog 11625019) were transfected with a pCMV-TP plasmid construct carrying nucleotides encoding human IL4Rα (amino acid residues 1-825 of uniprot #P24394-1) between EcoRI and XbaI, and a stable cell pool (designated 293F-IL4Rα) was selected for subsequent cell-based FACS binding and cell-based ligand blocking FACS analysis. 293F-IL4Rα cells were harvested from cell culture flasks, washed twice, and resuspended in FACS buffer (phosphate buffered saline (PBS) containing 2% v / v fetal bovine serum). Then, 2×10 5 Anti-IL4Rα antibodies or control substances serially diluted in FACS buffer (4-fold serial dilutions from a starting concentration of 80 nM) were added at 100 μL / well to a 96-well plate containing cells / well and incubated on ice for 40 min. Cells were washed twice with FACS buffer, and then R-phycoerythrin-labeled affinity-purified goat anti-human IgG (Fcγ fragment specificity, Jackson Immunoresearch, catalog 109-115-098, diluted 1:1000 in FACS buffer) was added at 100 μL / well. After 40 min incubation at 4°C in the dark, cells were washed three times and then resuspended in FACS buffer. Fluorescence values ​​were measured using a Becton Dickinson FACS CantoII-HTS. Data analysis was performed with the software Graphpad Prism, and EC 50 The value was calculated.

[0125] 2.3 Indirect ELISA The cross-reactivity of the anti-IL4Rα antibody of the present disclosure with cynomolgus monkey IL4Rα protein or cal-IL4Rα-his protein was analyzed. Briefly, cynomolgus monkey IL4Rα-his protein (Sino biological inc., catalog 90897-C08H) dissolved in carbonate / bicarbonate buffer (pH 9.6) at a concentration of 2 μg / mL or cal-IL4Rα-his protein (Sino biological inc. custom-made product, catalog BAX2) dissolved in carbonate / bicarbonate buffer (pH 9.6) at a concentration of 0.2 μg / mL was coated on a 96-well plate at 100 μL / well and incubated at 37° C. for 2 hours. The plate was washed once with washing buffer (PBS+0.05% w / v Tween-20, PBST), and then blocking buffer (PBST containing 5% w / v skim milk) was added at 200 μL / well and blocked for 2 hours at 37° C. The plate was washed again, and 100 μL of serially diluted (0.004-66.7 nM, 5-fold serial dilutions from a starting concentration of 66.7 nM in PBST containing 2.5% w / v skim milk) anti-IL4Rα antibodies of the present disclosure or control substances were added to each well and incubated at 37° C. for 40 minutes. After washing the plates four times, peroxidase-labeled affinity-purified F(ab')2 fragmented goat anti-human IgG (Fcγ fragment specificity, Jackson Immunoresearch, catalog 109-036-098, diluted 1:5000 in PBST buffer) was added at 100 μL / well and incubated for 40 min at 37°C. After the final wash, TMB (Innoreagents) was added at 100 μL / well and incubated. After 3–10 min, the reaction was stopped by adding 50 μL / well of 1 M H2SO4 at 25°C and the absorbance at 450 nm was measured. Data analysis was performed with the software Graphpad Prism, and EC 50 The value was calculated.

[0126] Example 3: Blocking activity of anti-IL4Rα antibodies against IL4Rα-reference substance or IL4Rα-IL4 interaction 3.1 Ligand blocking ELISA The ability of the anti-IL4Rα antibodies of the present disclosure to block IL4-IL4Rα interaction was confirmed by competitive ELISA. Briefly, human IL4Rα-his protein (prepared in-house, amino acid sequence as shown in SEQ ID NO: 15) dissolved in PBS at a concentration of 2 μg / mL was coated on a 96-well plate at 100 μL / well and incubated overnight at 4° C. The next day, the plate was washed with washing buffer (PBS+0.05% w / v Tween-20, PBST) and blocked by adding PBST containing 5% w / v skim milk at 37° C. for 2 hours. The plate was then washed again with washing buffer.

[0127] The anti-IL4Rα antibody of the present disclosure or a control substance was serially diluted (4-fold serial dilution from a starting concentration of 80 nM) in PBST buffer containing 2.5% w / v skim milk, and the serially diluted anti-IL4Rα antibody or control substance was added to the IL4Rα-coated plate at 100 μL / well and co-incubated with human IL4Rα-his protein for 40 minutes at 37° C. After washing the plate four times with washing buffer, 100 μL of biotin-labeled human IL4 protein (Sino biological inc., catalog 11846-HNAE) at a concentration of 0.56 nM was added to each well and incubated at 37° C. for 40 minutes. The plate was washed again with washing buffer. Then, 100 μL / well of HRP-conjugated streptavidin (diluted 1:10000 in PBST buffer, Jackson Immunoresearch, Cat. 016-030-084) was added and co-incubated for 40 min at 37°C. The plate was washed again with wash buffer. Finally, TMB was added, the reaction was stopped with 1M H2SO4, and the absorbance at 450 nm was measured. Data analysis was performed with the software Graphpad Prism, and IC 50 The value was calculated.

[0128] 3.2 Reference substance blocking ELISA The ability of the anti-IL4Rα antibody of the present disclosure to block reference substance-human IL4Rα binding was confirmed by competitive ELISA. Briefly, the reference substance dissolved in PBS at a concentration of 2 μg / mL was coated on a 96-well plate at 100 μL / well and incubated overnight at 4° C. The next day, the plate was washed with wash buffer (PBS+0.05% w / v Tween-20, PBST) and blocked for 2 hours at 37° C. by adding PBST containing 5% w / v skim milk. During blocking of the 96-well plate, the anti-IL4Rα antibody of the present disclosure or the control substance was diluted (4-fold serial dilution from a starting concentration of 100 nM) with biotin-labeled human IL4Rα-his protein (prepared in-house, amino acid sequence as shown in SEQ ID NO: 15, dissolved in PBST containing 2.5% w / v skim milk at a concentration of 0.55 nM) and incubated at 25° C. for 40 minutes. After washing the plate, the antibody / IL4Rα-his mixture was added to the reference coated plate at 100 μL / well. It was incubated at 37°C for 40 min and the plate was washed again with washing buffer. Next, 100 μL / well of HRP-labeled streptavidin was added to the plate and incubated at 37°C for 40 min to detect plate-bound biotin-labeled human IL4Rα-his. The plate was washed again with washing buffer. Finally, TMB was added, the reaction was stopped with 1M H2SO4, and the absorbance at 450 nm was measured. Data analysis was performed with the software Graphpad Prism and IC 50 The value was calculated.

[0129] 3.3 Cell-based ligand blockade FACS Using the 293F-IL4Rα cells prepared as described above, the activity of the anti-IL4Rα antibody to block the binding of IL4 protein to IL4Rα on the cell surface was evaluated by flow cytometry (FACS).

[0130] Briefly, 293F-IL4Rα cells were harvested from cell culture flasks, washed twice, and resuspended in FACS buffer (PBS containing 2% v / v fetal bovine serum). Then, 1 × 10 5100μL / well of anti-IL4Rα antibody or control substance serially diluted in FACS buffer (4-fold serial dilution from starting concentration of 80nM) was added to a 96-well plate containing cells / well and incubated on ice for 40 minutes. After washing the plate twice with FACS buffer, 100μL of biotin-labeled human IL4 protein (Sino biological inc., catalog 11846-HNAE) at a concentration of 1.67nM was added to each well and incubated at 4℃ in the dark for 40 minutes. After washing the plate twice with FACS buffer, 100μL of R-phycoerythrin-labeled streptavidin (diluted 1:500 in FACS buffer; Jackson Immunoresearch, catalog 016-110-084) was added to each well and incubated at 4℃ in the dark for 40 minutes. After washing the plate twice with FACS buffer, 100μL of R-phycoerythrin-labeled streptavidin (diluted 1:500 in FACS buffer; Jackson Immunoresearch, catalog 016-110-084) was added to each well and incubated at 4℃ in the dark for 40 minutes. After washing the cells twice, they were resuspended in FACS buffer. Fluorescence values ​​were measured using a Becton Dickinson FACS CantoII-HTS. Data were analyzed using the software Graphpad Prism, and IC 50 The value was calculated.

[0131] Example 4: Cell-Based Functional Assay of Anti-IL4Rα Antibodies IL4 and IL13 can bind to IL4Rα and induce STAT6 phosphorylation in HEK293T-IL4Rα-STAT6-STAT6LUC-LB2 cells. STAT6 phosphorylation is crucial in the IL4 / IL13 signaling pathway.

[0132] Briefly, we stably transfected HEK293T cells (ATCC CRL-11268) naturally expressing IL13Rα1 with pcDNA3.1-Puro (YouBio biological inc., Catalog VT9222) plasmid construct (containing nucleotides encoding human IL4Rα between BamHI and XhoI), STAT6 plasmid (Sino biological inc., Catalog HG13190-NH) (containing nucleotides encoding human STAT6 between KpnI and XbaI), and STAT6 luciferase reporter plasmid STAT6-Luc (Yeasen biological inc., Catalog 11588ES03) to prepare HEK293T-IL4Rα-STAT6-STAT6LUC-LB2 cells in-house. We then selected a single cell clone LB2 for all subsequent functional assays.

[0133] The inhibitory effect of the anti-IL4Rα antibodies of the present disclosure on STAT6 phosphorylation induced by IL4 and IL13 was analyzed.

[0134] Briefly, log-phase HEK293T-IL4Rα-STAT6-STAT6LUC-LB2 cells were resuspended in culture medium (RPMI1640 + 10% FBS) and plated at 5 × 10 cells per well. 5Cells were seeded in a 96-well plate at 100 μL / well. Then, 50 μL of serially diluted (5-fold serial dilutions from a starting concentration of 100 nM) anti-IL4Rα antibodies of the present disclosure or control substances (including anti-CD22 antibodies prepared in-house) were added to each well and incubated at 37° C. for 30 minutes. Then, 50 μL of IL4 protein (600 pg / mL, Sino biological inc., catalog 11846-HNAE) or IL13 protein (50 ng / mL, Sino biological inc., catalog 10369-HNAC) was added to each well and incubated at 37° C. for 20 minutes. The plate was centrifuged and washed twice with staining buffer (prepared in-house, DPBS + 0.5% w / v BSA + 2mM EDTA), after which 50μL of fixation buffer (BD biosciences inc., catalog 5545655) was added to each well and incubated at 4℃ for 30 minutes. The cells were washed twice, and 200μL of permeabilization buffer (BD biosciences inc., part number 558050) was added to each well and incubated on ice for 30 minutes. The plate was washed three times with staining buffer. Then, anti-pSTAT6 antibody (pSTAT6 stock solution 20 times diluted, BD biosciences inc., catalog 562079) was added and incubated on ice for 60 minutes. Finally, the plate was washed twice and resuspended in staining buffer. Fluorescence values ​​were measured using Becton Dickinson FACS CantoII-HTS. Data analysis was performed using the software Graphpad Prism, and IC 50 The value was calculated.

[0135] Example 5: Production and analysis of chimeric antibodies The heavy and light chain variable regions of the mouse anti-IL4Rα monoclonal antibody were sequenced. The sequence numbers are summarized in Table 1.

[0136] The heavy chain variable region and light chain variable region of the mouse anti-IL4Rα monoclonal antibody C2C1A1A1 were cloned into a vector containing a human IgG4 heavy chain constant region (sequence number 13) and a vector containing a human κ light chain constant region (sequence number 14), respectively, such that the C-terminus of the variable region was linked to the N-terminus of the corresponding constant region.

[0137] A vector containing nucleotides encoding a heavy chain variable region linked to a human IgG4 heavy chain constant region and a vector containing nucleotides encoding a light chain variable region linked to a human kappa light chain constant region were transiently transfected into 50 mL of 293F suspension cells with 1 mg / mL PEI at a ratio of 60%:40% light chain construct:heavy chain construct. The cell supernatant was harvested after 6 days of culture in shake flasks, and the cells in the supernatant were sedimented by centrifugation and filtered through a 0.22 μm filter for isolation of immunoglobulins. The chimeric antibodies were purified by protein A affinity chromatography. Briefly, a protein A agarose column (bestchrom (Shanghai) Biosciences, catalog AA0273) was washed with 5–10 column volumes of PBS buffer. After the cell supernatant was passed through the protein A agarose column, the column was washed with PBS buffer until the absorbance of the protein returned to baseline. The eluate eluted from the column with elution buffer (0.1 M glycine-HCl, pH 2.7) was immediately collected in a 1.5 mL tube and neutralized with neutralization buffer (1 M Tris-HCl, pH 9.0). The fractions containing immunoglobulins were pooled and dialyzed in PBS at 4 °C overnight.

[0138] The purified antibodies were analyzed by capture ELISA, competitive ELISA, BIAcore affinity assay, cell-based FACS binding assay, and cell-based functional assay according to the protocols in the above examples. Meanwhile, for mouse C2C1A1A1, the analysis method was adjusted as follows in the above examples.

[0139] For the capture ELISA, 2 μg / mL goat anti-mouse IgG (Fcγ fragment specific, Jackson Immunoresearch, Cat 115-005-008) was added at 100 μL / well in place of goat anti-human IgG.

[0140] For BIAcore, goat anti-mouse IgG (GE healthcare, catalog BR100838, Mouse Antibody Capture Kit) was covalently coupled to a CM5 chip instead of goat anti-human IgG.

[0141] For cell-based FACS binding, R-phycoerythrin-labeled affinity-purified F(ab')2 fragmented goat anti-mouse IgG (Jackson Immunoresearch, catalog 115-116-146) was added at 100 μL / well instead of R-phycoerythrin-labeled affinity-purified goat anti-human IgG.

[0142] The results are shown in Table 2. The data showed that the chimeric antibodies had similar binding affinity / capacity and blocking activity as the parental murine antibodies.

[0143] [Table 2]

[0144] Example 6: Humanization of anti-IL4Rα monoclonal antibody The murine anti-IL4Rα antibody C2C1A1A1 was humanized and characterized. The murine antibody was humanized using established CDR grafting methods as follows.

[0145] To select an acceptor framework for humanizing the mouse antibody C2C1A1A1, the sequences of the light and heavy chain variable regions of each mouse antibody were BLASTed against the human immunoglobulin gene database. The human germline antibody with the highest homology was selected as the acceptor framework for humanization. The heavy / light chain variable region CDRs of the mouse antibody were inserted into the selected framework, and more candidate heavy / light chain variable regions were obtained by backmutating residues in the framework. A total of 16 exemplary humanized C2C1A1A1 antibodies, huC2C1A1A1-V1 to huC2C1A1A1-V16, were obtained. The sequence numbers of these heavy / light chain variable regions are shown in Table 1.

[0146] A vector containing nucleotides encoding a humanized heavy chain variable region linked to a human IgG4 heavy chain constant region (SEQ ID NO: 13) and a vector containing nucleotides encoding a humanized light chain variable region linked to a human kappa light chain constant region (SEQ ID NO: 14) were transiently transfected into 50 mL of 293F suspension cells with 1 mg / mL PEI at a ratio of 60%:40% light chain construct:heavy chain construct.

[0147] The cell supernatant was harvested after 6 days of culture in shake flasks, and the cells in the supernatant were sedimented by centrifugation and filtered through a 0.22 μm filter for isolation of immunoglobulins. The antibodies were purified by protein A affinity chromatography. Briefly, a protein A agarose column (bestchrom(Shanghai)Biosciences, catalog AA0273) was washed with 5–10 column volumes of PBS buffer. After the cell supernatant was passed through the protein A agarose column, the column was washed with PBS buffer until the absorbance of the protein returned to baseline. The eluate eluted from the column with elution buffer (0.1 M glycine-HCl, pH 2.7) was immediately collected in a 1.5 mL tube and neutralized with neutralization buffer (1 M Tris-HCl, pH 9.0). The fractions containing immunoglobulins were pooled and dialyzed in PBS at 4 °C overnight.

[0148] Example 7. Characterization of humanized antibodies According to the protocol of the above example, the binding affinity of the humanized antibody to human IL4Rα was evaluated by BIAcore technology, and the K a , K d and K D The results are summarized in Table 3.

[0149] [Table 3]

[0150] The results show that the humanized antibodies have similar human IL4Rα binding affinity to the chimeric antibodies, and all humanized huC2C1A1A1 antibodies showed higher human IL4Rα binding affinity compared to the reference.

[0151] Following the protocols of the above examples, humanized antibodies huC2C1A1A1-V14 and huC2C1A1A1-V15 were further analyzed by Biacore, capture ELISA, indirect ELISA, cell-based FACS binding, competitive ELISA, cell-based ligand blocking FACS and cell-based functional assays.

[0152] We also analyzed the thermal stability of the humanized antibody huC2C1A1A1-V15. (商標) The Tm (melting temperature) was measured by protein thermal shift assay using the Thermal Shift Protein Stability Kit (Biotium, Cat. 33022-T, Lot No. 181214). (商標)The dye was thawed to room temperature and the vial containing the dye was vortexed and centrifuged. 5 μL of 200× dye was then added to 95 μL of PBS to prepare 10× dye. 2 μL of 10× dye and 10 μg of humanized antibody were added to the reaction and PBS was added until the total reaction volume was 20 μL. The tube containing the dye and antibody was centrifuged briefly and placed in a real-time PCR thermal cycler (Roche, LightCycler 480II) with the program MeltCurve set with the parameters in Table 4. The results are shown in Tables 5-1, 5-2 and 5-3. The data show that the humanized C2C1A1A1 antibody exhibited comparable (if not better) human IL4Rα binding affinity / activity and blocking ability against IL4Rα-IL4 / IL13 compared to the reference material.

[0153] [Table 4]

[0154] [Table 5]

[0155] [Table 6]

[0156] [Table 7]

[0157] Example 8: Preparation and screening of pharmaceutical compositions [Measurement method] (1) Size Exclusion Chromatography (SEC-UPLC) To measure the purity of the antibody in the sample, high performance liquid chromatography was performed using a Thermo Vanquish F with a Waters ACQUITY UPLC Protein BEH SEC column (200 Å, 1.7 μm, 4.6 mm × 300 mm) and a Waters ACQUITY UPLC Protein BEH SEC guard column (200 Å, 1.7 μm, 4.6 mm × 30 mm) as a precolumn, elution was performed with 50 mmol / L phosphate buffer + 200 mmol / L sodium chloride (pH 7.0) as the mobile phase, and the measurement wavelength was set at 280 nm. The content of polymeric impurities and immunoglobulin monomers was calculated by the area normalization method.

[0158] (2) Differential scanning fluorescence (DSF) The unfolding temperature (Tm) of the sample was measured using a protein stability analyzer (Nano Temper, Prometheus NT.48). 10 μl of the sample was placed in the sample chamber using a capillary tube. For the process, the scan start temperature was 25°C, the scan end temperature was 95°C, and the heating rate was 0.3°C / min.

[0159] (3) The viscosity of the samples was analyzed using a HVROC-S viscometer (Rheosense, μVISC), the automatic mode was selected, and the measurement results were recorded.

[0160] (4) Whole-column imaging capillary isoelectric focusing (iCIEF) To identify the charge variants of the samples, we performed detection at a UV detection wavelength of 280 nm using an imaging capillary isoelectric focusing apparatus (Protein Simple iCE3), a fast and fully automated protein characterization system (Protein Simple, Maurice), a quartz capillary tube with a protective layer (Protein Simple), and a UV detector. The injection time was set to 60 seconds, and the samples were prefocused at 1500 volts for 1 minute and focused at 300 volts for 10 minutes. The peak area percentage of each charge variant was calculated by the peak area normalization method.

[0161] [Preparation of buffer] (1) Histidine-histidine hydrochloride buffer contains histidine and histidine hydrochloride. In a specific embodiment, the histidine-histidine hydrochloride buffer contains histidine and histidine hydrochloride monohydrate, for example, 20 mM histidine-histidine hydrochloride buffer (pH 5.8) contains about 1.17 mg / mL of histidine and about 2.62 mg / mL of histidine hydrochloride monohydrate.

[0162] (2) Sodium phosphate buffer includes disodium hydrogen phosphate and sodium dihydrogen phosphate. In a specific embodiment, the sodium phosphate buffer includes sodium dihydrogen phosphate monohydrate and disodium hydrogen phosphate dodecahydrate, for example, 20 mM sodium phosphate buffer (pH 6.0) is formed from about 2.539 mg / mL of sodium dihydrogen phosphate monohydrate and about 0.573 mg / mL of disodium hydrogen phosphate dodecahydrate.

[0163] (3) Sodium acetate-acetic acid buffer comprises sodium acetate and acetic acid. In a specific embodiment, the sodium acetate-acetic acid buffer comprises sodium acetate trihydrate and acetic acid, for example, 20 mM sodium acetate-acetic acid buffer (pH 6.0) is formed from about 2.605 mg / mL sodium acetate trihydrate and about 0.141 mg / mL acetic acid.

[0164] Anti-IL4Rα antibody (huC2C1A1A1-V15) was substituted with each of the buffers in Table 6 by ultrafiltration, and after the substitution, the mixture was concentrated, and then surfactants and stabilizers were added as shown in Table 6. After homogeneous mixing, the mixture was sterilized by filtration through a 0.22 μm filter, dispensed into vials, and then stoppered and capped. The SEC-UPLC measurement results of pharmaceutical compositions F1 to F4 are shown in Table 7. Among them, F4 showed a greater increase in polymeric impurities (increase from 1.36% to 5.66%) after storage at 40°C for 1 month than F1 to F3.

[0165] [Table 8]

[0166] [Table 9]

[0167] As an example, 20 mM histidine-histidine hydrochloride was selected as a buffer. Anti-IL4Rα antibody (huC2C1A1A1-V15) was replaced with the buffer in Table 8 by ultrafiltration, and after the replacement, the mixture was concentrated, and then surfactants and stabilizers were added as shown in Table 8. After uniform mixing, the mixture was filtered through a 0.22 μm filter for sterilization, dispensed into vials, and then stoppered and capped. Table 9 shows the Tm and SEC-UPLC measurement results for pharmaceutical compositions F5 to F7. Among them, F6 showed the smallest increase in polymeric impurities after storage at 40° C. for 1 month.

[0168] [Table 10]

[0169] [Table 11]

[0170] As an example, 20 mM histidine-histidine hydrochloride was selected as a buffer. Anti-IL4Rα antibody (huC2C1A1A1-V15) was replaced with the buffer in Table 10 by ultrafiltration, and after the replacement, the mixture was concentrated, and then surfactants and stabilizers were added as shown in Table 10. After uniform mixing, the mixture was sterilized by filtering through a 0.22 μm filter, dispensed into vials, and then stoppered and capped. The Tm, viscosity, SEC-UPLC and iCIEF measurement results of pharmaceutical compositions F8 and F9 are shown in Table 11. Among them, F8 had a lower viscosity than F9.

[0171] [Table 12]

[0172] [Table 13]

[0173] While this disclosure has been described with reference to one or more embodiments, it is to be understood that the disclosure is intended to encompass not only those embodiments, but also all alternatives, modifications, and equivalents falling within the spirit and scope of the appended claims. All documents referenced herein are hereby incorporated by reference. The sequence information of this disclosure is summarized in Table 12 below.

[0174] [Table 14-1]

[0175] [Table 14-2]

[0176] [Table 14-3]

Claims

1. A pharmaceutical composition comprising: (a) an anti-IL4Rα antibody or an antigen-binding fragment thereof; (b) a buffer; (c) a surfactant; and (d) a stabilizer.

2. The pharmaceutical composition of claim 1, wherein the anti-IL4Rα antibody or antigen-binding fragment thereof comprises a heavy chain variable region CDR1, a heavy chain variable region CDR2, a heavy chain variable region CDR3, a light chain variable region CDR1, a light chain variable region CDR2, and a light chain variable region CDR3, and the heavy chain variable region CDR1, the heavy chain variable region CDR2, the heavy chain variable region CDR3, the light chain variable region CDR1, the light chain variable region CDR2, and the light chain variable region CDR3 each comprise an amino acid sequence that is at least 85% identical to the amino acid sequences represented by SEQ ID NOs: 1, 2, 3, 4, 5, and 6, respectively.

3. The anti-IL4Rα antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising an amino acid sequence having at least 85% identity to the amino acid sequence represented by SEQ ID NO: 7, 8, or 9, wherein the amino acid sequence of SEQ ID NO: 8 is EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYGMSWVRQAPGKGLVX １ VX ２ TINSNGGSTSYPDSVKGRFTISRDNAKNTLYLQMNSLRAEDTAVYYCARFFRFRNAMDYWGQGTLVTVSS (X in SEQ ID NO: 8 １ = W and X ２ = S or X １ = L and X ２ = A or X １ = W and X ２ = A) and / or The anti-IL4Rα antibody or antigen-binding fragment thereof comprises a light chain variable region comprising an amino acid sequence having at least 85% identity to the amino acid sequence represented by SEQ ID NO: 10, 11, or 12, wherein the amino acid sequence of SEQ ID NO: 11 is DIQMTQSPSSLSASVGDRVTITCRTSENIYSYLAWYQQKPGKAPKX １ LX ２ YNAKTLAEGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQHYYGPPTWTFGQGTKVEIK (X in SEQ ID NO: 11 １ = L and X ２ = I or X １ = F and X ２ = V or X １ = F and X ２ 3. The pharmaceutical composition according to claim 1 or 2, wherein:

4. The anti-IL4Rα antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, (1) an amino acid sequence having at least 85% identity with the amino acid sequences represented by SEQ ID NOs: 7 and 10, respectively; (2) an amino acid sequence having at least 85% identity with the amino acid sequences represented by SEQ ID NOs: 8 and 11, respectively (X in SEQ ID NO: 8) １ = W and X ２ = S), (3) an amino acid sequence having at least 85% identity with the amino acid sequences represented by SEQ ID NOs: 8 and 12, respectively (X in SEQ ID NO: 8) １ = W and X ２ = S), (4) an amino acid sequence having at least 85% identity with the amino acid sequences represented by SEQ ID NOs: 9 and 11, respectively; (5) an amino acid sequence having at least 85% identity with the amino acid sequences represented by SEQ ID NOs: 9 and 12, respectively; (6) an amino acid sequence having at least 85% identity with the amino acid sequences represented by SEQ ID NOs: 8 and 11, respectively (X in SEQ ID NO: 8) １ = L and X ２ = A), (7) an amino acid sequence having at least 85% identity with the amino acid sequences represented by SEQ ID NOs: 8 and 12, respectively (X in SEQ ID NO: 8) １ = L and X ２ = A), (8) an amino acid sequence having at least 85% identity with the amino acid sequences represented by SEQ ID NOs: 8 and 11, respectively (X in SEQ ID NO: 8) １ = W and X ２ = A), or (9) an amino acid sequence having at least 85% identity with the amino acid sequences represented by SEQ ID NOs: 8 and 12, respectively (X in SEQ ID NO: 8) １ = W and X ２ = A), Here, the amino acid sequence of SEQ ID NO: 8 is EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYGMSWVRQAPGKGLVX １ VX ２ TINSNGGSTSYPDSVKGRFTISRDNAKNTLYLQMNSLRAEDTAVYYCARFFRFRNAMDYWGQGTLVTVSS, The amino acid sequence of SEQ ID NO: 11 is DIQMTQSPSSLSASVGDRVTITCRTSENIYSYLAWYQQKPGKAPKX １ LX ２ YNAKTLAEGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQHYYGPPTWTFGQGTKVEIK (X in SEQ ID NO: 11 １ = L and X ２ = I, X １ = F and X ２ = V or X １ = F and X ２ = I), The pharmaceutical composition according to any one of claims 1 to 3.

5. The pharmaceutical composition according to any one of claims 1 to 4, wherein the anti-IL4Rα antibody or antigen-binding fragment thereof further comprises a heavy chain constant region comprising the amino acid sequence represented by SEQ ID NO: 13, and a light chain constant region comprising the amino acid sequence represented by SEQ ID NO:

14.

6. The pharmaceutical composition according to any one of claims 1 to 5, wherein the concentration of the anti-IL4Rα antibody or antigen-binding fragment thereof is 30 mg / mL to 300 mg / mL, 50 mg / mL to 250 mg / mL, 70 mg / mL to 200 mg / mL, 100 mg / mL to 180 mg / mL, 120 mg / mL to 180 mg / mL, 120 mg / mL to 150 mg / mL, or 150 mg / mL to 180 mg / mL.

7. The pharmaceutical composition of any one of claims 1 to 6, wherein the buffer comprises a phosphate buffer, an acetate buffer, or a histidine buffer.

8. The pharmaceutical composition described in claim 7, wherein the phosphate buffer is a sodium phosphate buffer, the acetate buffer is a sodium acetate-acetic acid buffer, and / or the histidine buffer is a histidine-histidine hydrochloride buffer.

9. A pharmaceutical composition described in any one of claims 1 to 8, wherein the concentration of the buffering agent is 1 mM to 100 mM, 2 mM to 80 mM, 4 mM to 60 mM, 8 mM to 40 mM, 10 mM to 30 mM, 10 mM to 20 mM, or 20 mM to 30 mM.

10. The pharmaceutical composition of any one of claims 1 to 9, wherein the surfactant comprises polysorbate 80 or polysorbate 20.

11. A pharmaceutical composition described in any one of claims 1 to 10, wherein the concentration of the surfactant is 0.01 mg / mL to 2 mg / mL, 0.05 mg / mL to 1 mg / mL, 0.1 mg / mL to 0.8 mg / mL, 0.2 mg / mL to 0.6 mg / mL, or 0.2 mg / mL to 0.4 mg / mL.

12. The pharmaceutical composition according to any one of claims 1 to 11, wherein the stabilizer is selected from trehalose, mannitol, sucrose, arginine or a pharmaceutically acceptable salt thereof, proline, or sodium chloride.

13. The concentration of the sucrose is 10 mg / mL to 100 mg / mL, 20 mg / mL to 80 mg / mL, 30 mg / mL to 60 mg / mL, 40 mg / mL to 50 mg / mL, or 50 mg / mL to 60 mg / mL, the concentration of the arginine or a pharmaceutically acceptable salt thereof is 10 mM to 100 mM, 20 mM to 80 mM, 30 mM to 60 mM, 40 mM to 50 mM, or 50 mM to 60 mM, and the concentration of the proline is 10 mM to 100 mM, 20 mM to 80 mM, 30 mM to 60 mM, 40 mM to 50 mM, or 50 mM to 60 mM. and / or the concentration of sodium chloride is 1 mM to 80 mM, 2 mM to 60 mM, 4 mM to 50 mM, 8 mM to 40 mM, 10 mM to 30 mM, or 10 mM to 20 mM.

14. 14. The pharmaceutical composition according to any one of claims 1 to 13, wherein the pH of the pharmaceutical composition is 4 to 7, 5 to 7, 5 to 6.5, 5.3 to 6.5, 5.3 to 6.3, 5.3 to 5.8, or 5.8 to 6.

3.

15. The pharmaceutical composition comprises: (a) 30 mg / mL to 300 mg / mL of an anti-IL4Rα antibody or antigen-binding fragment thereof; (b) 1 mM to 100 mM of a buffering agent; (c) 0.01 mg / mL to 2 mg / mL of a surfactant; and (d) a stabilizer comprising trehalose, mannitol, sucrose, arginine or a pharmaceutically acceptable salt thereof, proline, and / or sodium chloride, wherein the pH of the pharmaceutical composition is 4 to 7, 5 to 7, 5 to 6.5, 5.3 to 6.5, 5.3 to 6.3, 5.3 to 5.8, or 5.8 to 6.3; or 15. The pharmaceutical composition according to any one of claims 1 to 14, comprising: (a) 30 mg / mL to 300 mg / mL of an anti-IL4Rα antibody or antigen-binding fragment thereof; (b) 1 mM to 100 mM of a phosphate buffer, acetate buffer, or histidine buffer; (c) 0.01 mg / mL to 2 mg / mL of polysorbate 80 or polysorbate 20; and (d) 10 mg / mL to 100 mg / mL of sucrose, 10 mM to 100 mM of arginine or a pharmaceutically acceptable salt thereof, 60 mM to 600 mM of proline, and / or 1 mM to 80 mM of sodium chloride, wherein the pH of the pharmaceutical composition is 4 to 7, 5 to 7, 5 to 6.5, 5.3 to 6.5, 5.3 to 6.3, 5.3 to 5.8, or 5.8 to 6.

3.

16. 16. A lyophilized formulation comprising an anti-IL4Rα antibody or antigen-binding fragment thereof, wherein the lyophilized formulation is obtained by lyophilizing the pharmaceutical composition of any one of claims 1 to 15, or is capable of being dissolved to form the pharmaceutical composition of any one of claims 1 to 15.

17. Use of an anti-IL4Rα antibody or an antigen-binding fragment thereof for producing a pharmaceutical composition according to any one of claims 1 to 15 or a freeze-dried preparation according to claim 16 for the treatment of an allergic disease.

18. The use described in claim 17, wherein the allergic disease is atopic dermatitis, allergic reaction, allergic rhinitis or allergic asthma.

19. Use of an anti-IL4Rα antibody or an antigen-binding fragment thereof for producing the pharmaceutical composition of any one of claims 1 to 15 or the lyophilized formulation of claim 16 for treating a tumor associated with increased STAT6 activation in a subject.

20. The use described in claim 19, wherein the tumor is a solid tumor.

21. The use of claim 19, wherein the tumor is melanoma, lung cancer, renal cancer, prostate cancer, cervical cancer, colorectal cancer, gastric cancer, pancreatic cancer, ovarian cancer and / or urothelial cancer.

22. Use of an anti-IL4Rα antibody or an antigen-binding fragment thereof for producing the pharmaceutical composition of any one of claims 1 to 15 or the lyophilized formulation of claim 16 for reducing IL4 and / or IL13 signaling in a subject or a type 2 immune response in a subject.

23. The use described in claim 22, wherein the IL4 and / or IL13 signaling is manifested as activation and / or proliferation of B cells, eosinophils, macrophages, proliferation of fibroblasts and proliferation of smooth skin.

24. The use described in claim 23, wherein the proliferation of smooth muscle cells is proliferation of airway smooth muscle cells.