Stable antibody composition

Abstract

The present invention relates to an aqueous pharmaceutical composition comprising risankizumab or an antigen-binding fragment thereof; and a stabilizer, and not containing a polyol, a method for producing the same, and uses thereof.

Description

[Technical Field]

[0001] The present invention relates to stable pharmaceutical compositions comprising risankizumab or an antigen-binding fragment thereof, methods for their preparation, and pharmaceutical uses thereof for the treatment of various diseases. [Background technology]

[0002] Risankizumab is a highly effective and specific inhibitor of IL-23. It is a humanized immunoglobulin G1 (IgG1) monoclonal antibody directed against the p19 subunit of IL-23. By binding to IL-23p19, risankizumab inhibits the function of IL-23 to induce and maintain T helper (Th)17 cells, innate lymphoid cells, gamma delta T cells, and natural killer (NK) cells, which are responsible for tissue inflammation, destruction, and abnormal tissue repair. Risankizumab is known to be effective in treating autoimmune diseases, specifically inflammatory bowel disease (e.g., Crohn's disease, ulcerative colitis), multiple sclerosis, rheumatoid arthritis, ankylosing spondylitis, psoriatic arthritis, and psoriasis (see WO2012 / 061448).

[0003] For effective administration of risankizumab, a pharmaceutical formulation with excellent stability is required. WO2021 / 048743 discloses a liquid pharmaceutical formulation containing risankizumab as well as a polyol and a surfactant as stabilizers. However, polyols, such as sugars or sugar alcohols, are known to contain monosaccharides as impurities, and polyols can decompose into monosaccharides under pH and temperature stress conditions (The effect of sucrose hydrolysis on the stability of protein therapeutics during accelerated formulation studies. Journal of Pharmaceutical Sciences, 2009 December;98(12):4501-10). However, monosaccharides can react with the amino acid groups of proteins to induce glycation, which is known to pose potential risks, such as causing protein denaturation (e.g., aggregation) or affecting protein efficacy (Glycation of polyclonal IgGs: Effect of sugar excipients during stability studies, European Journal of Pharmaceutics and Biopharmaceutics, Volume 102, May 2016, Pages 185-190; Quantitative analysis of glycation and its impact on antigen binding, MAbs. 2018, Volume 10, No. 3, 406-415). [Prior art documents] [Patent documents]

[0004] [Patent Document 1] WO2012 / 061448 [Patent Document 2] WO2021 / 048743 [Non-patent literature]

[0005] [Non-Patent Document 1] The effect of sucrose hydrolysis on the stability of protein therapeutics during accelerated formulation studies. Journal of pharmaceutical sciences, 2009 Dec;98(12):4501-10 [Non-patent document 2] Quantitative analysis of glycation and its impact on antigen binding, MAbs. 2018, Volume 10, No.3, 406-415 Summary of the Invention [Problem to be solved by the invention]

[0006] Therefore, there is a need to develop a risankizumab formulation that has a stabilizer that can replace polyol and further improve the stability of risankizumab.

[0007] One aspect of the present invention is to provide a stable pharmaceutical composition suitable for use as a medicament for the treatment of an individual comprising risankizumab or an antigen-binding fragment thereof.

[0008] Another aspect of the present invention is to provide a method of treating an autoimmune disease in an individual comprising administering to the individual the pharmaceutical composition described above.

[0009] Yet another aspect of the present invention is to provide a method for preparing said pharmaceutical composition. [Means for solving the problem]

[0010] One aspect of the present invention is (a) risankizumab or an antigen-binding fragment thereof; and (b) contains a stabilizer; A polyol-free aqueous pharmaceutical composition is provided.

[0011] Another aspect of the present invention is (a) risankizumab or an antigen-binding fragment thereof; and (b) an amino acid or a pharmaceutically acceptable salt thereof; (c) providing an aqueous pharmaceutical composition having a pH of 5.0 to 7.0;

[0012] Yet another aspect of the present invention provides a method of treating an autoimmune disease comprising administering to an individual the pharmaceutical composition described above. [Effects of the Invention]

[0013] It has been found that an aqueous pharmaceutical composition comprising risankizumab or an antigen-binding fragment thereof according to one embodiment of the present invention does not contain a polyol, which has been conventionally used as a stabilizer, but contains an amino acid or its salt or metal salt, such as sodium chloride, as a stabilizer, and thereby has excellent stability under various conditions, including heat stability, light stability, freezing and / or cold-thawing stability, and agitation stability. Furthermore, because the composition has excellent stability with or without a surfactant and / or buffer, it is possible to avoid side effects that may be caused by the inclusion of a surfactant and / or buffer. [Brief explanation of the drawings]

[0014] [Figure 1] FIG. 1 shows the amino acid sequence of the light chain of risankizumab (SEQ ID NO: 1). [Figure 2] FIG. 1 shows the amino acid sequence of the heavy chain of risankizumab (SEQ ID NO: 2). [Figure 3] 1 is a graph showing the change in HMW% (ΔHMW%) at the initial 1, 2, and 4 weeks for each stabilizer obtained in a thermal stability test for risankizumab formulations containing only stabilizers, without buffer or surfactant. [Figure 4]Five factors (protein concentration, pH, buffer histidine concentration, stabilizer proline concentration, and surfactant concentration) of a risankizumab formulation containing proline as a stabilizer were designed based on DoE (Design of Experiments), and stability modeling was performed on the results (SE-HPLC and WCX results) of a thermal stability test. This graph was obtained by performing stability modeling using DoE statistical analysis. [Figure 5] Five factors (protein concentration, pH, buffer histidine concentration, stabilizer proline concentration, and surfactant concentration) of a risankizumab formulation containing proline as a stabilizer were designed based on DoE (Design of Experiments), and stability modeling was performed on the results (SE-HPLC and WCX results) of a thermal stability test. This graph was obtained by performing stability modeling using DoE statistical analysis. [Figure 6] Five factors (protein concentration, pH, buffer histidine concentration, stabilizer proline concentration, and surfactant concentration) of a risankizumab formulation containing proline as a stabilizer were designed based on DoE (Design of Experiments), and stability modeling was performed on the results (SE-HPLC and WCX results) of a thermal stability test. This graph was obtained by performing stability modeling using DoE statistical analysis. [Figure 7] Five factors (protein concentration, pH, buffer histidine concentration, stabilizer proline concentration, and surfactant concentration) of a risankizumab formulation containing proline as a stabilizer were designed based on DoE (Design of Experiments), and stability modeling was performed on the results (SE-HPLC and WCX results) of a thermal stability test. This graph was obtained by performing stability modeling using DoE statistical analysis. DETAILED DESCRIPTION OF THE INVENTION

[0015] Unless otherwise specified, all technical terms used herein have the same meaning as commonly understood by those skilled in the art. Furthermore, although preferred methods and samples are described herein, similar or equivalent methods and samples are also included within the scope of the present invention. Furthermore, numerical values ​​described herein are considered to include the meaning of "about" even if not explicitly stated. The contents of all publications referenced herein are incorporated herein by reference in their entirety.

[0016] The present invention, in one aspect, comprises: (a) risankizumab or an antigen-binding fragment thereof; and (b) contains a stabilizer; A polyol-free aqueous pharmaceutical composition is provided.

[0017] The stabilizer comprises an amino acid or a pharmaceutically acceptable salt or metal salt thereof. The metal salt can be NaCl, KCl, NaF, KBr, NaBr, Na2SO4, NaSCN, CaCl2, MgCl2, or K2SO4. The metal salt can be, for example, NaCl or Na2SO4. The metal salt is present in the pharmaceutical composition at a concentration of 0.5 to 1 wt%. In one specific example, the pharmaceutical composition contains 0.8 wt% sodium chloride. The concentration of the metal salt can be freely adjusted within a range that maintains the stability of risankizumab or its antigen-binding fragment in the pharmaceutical composition and can vary individually depending on the specific metal salt.

[0018] The present invention, in another aspect, comprises (a) risankizumab or an antigen-binding fragment thereof; and (b) an amino acid or a pharmaceutically acceptable salt thereof; (c) The aqueous pharmaceutical composition has a pH of 5.0 to 7.0. The pharmaceutical composition of this embodiment may be free of polyol.

[0019] The above two aspects of the pharmaceutical composition will be explained in more detail below.

[0020] The amino acid acts as a stabilizer and includes, but is not limited to, lysine, arginine, glycine, proline, histidine, alanine, valine, leucine, isoleucine, proline, phenylalanine, tyrosine, tryptophan, serine, threonine, cysteine, methionine, asparagine, glutamine, aspartic acid, glutamic acid, or a pharmaceutically acceptable salt thereof, or a mixture thereof. In one embodiment, the amino acid is lysine, arginine, glycine, proline, histidine, or a pharmaceutically acceptable salt thereof, or any combination thereof. In one embodiment, the amino acid is proline.

[0021] As the stabilizer, the concentration of amino acid is 0.1 to 300.0 mM, 0.5 to 300.0 mM, 1.0 to 300.0 mM, 5.0 to 300.0 mM, 10.0 to 300.0 mM, 25.0 to 300.0 mM, 30.0 to 300.0 mM, 50.0 to 300.0 mM, 80.0 to 300.0 mM, 100.0 to 300.0 mM, 120.0 to 300.0 mM, 0.1 to 250.0 mM, 0.5 to 250.0 mM, 1.0 to 250.0 mM, 5.0 to 250.0 mM, 10.0 to 250.0 mM M, 25.0 to 250.0 mM, 30.0 to 250.0 mM, 50.0 to 250.0 mM, 80.0 to 250.0 mM, 100.0 to 250.0 mM, 120.0 to 250.0 mM, 0.1 to 200.0 mM, 0.5 to 200.0 mM, 1.0 to 200.0 mM, 5.0 to 200.0 mM, 10.0 to 200.0 mM, 25.0 to 200.0 mM, 30.0 to 200.0 mM, 50.0 to 200.0 mM, 80.0 to 200.0 mM, 100.0 to 200.0 mM, 120.0 to 2 00.0mM, 0.1 to 160.0mM, 0.5 to 160.0mM, 1.0 to 160.0mM, 5.0 to 160.0mM, 10.0 to 160.0mM, 25.0 to 160.0mM, 30.0 to 160.0mM, 50.0 to 160.0mM, 80.0 to 160.0mM, 100.0 to 160.0mM, 120.0 to 160.0mM, 130.0 to 150.0mM, 0.1 to 100.0mM, 0.5 to 100.0mM, 1.0 to 100.0mM, 5.0 to 100.0mM, 10.0 to 1 00.0mM, 25.0 to 100.0mM, 30.0 to 100.0mM, 50.0 to 100.0mM, 80.0 to 100.0mM, 0.1 to 50.0mM, 0.5 to 50.0mM, 1.0 to 50.0mM, 5.0 to 50.0mM, 10.0 to 50.0mM, 25.0 to 50.0mM, 30.0 to 50.0mM, 0.1 to 40.0mM, 0.5 to 40.0mM, 1.0 to 40.0mM, 5.0 to 40.0mM, 10.0 to 40.0mM, 25.0 to 40.0mM, 30.0 to 40.The amino acid concentration is 0 mM, 0.1 to 30.0 mM, 0.5 to 30.0 mM, 1.0 to 30.0 mM, 5.0 to 30.0 mM, 10.0 to 30.0 mM, 25.0 to 30.0 mM, 0.1 to 20.0 mM, 0.5 to 20.0 mM, 1.0 to 20.0 mM, 5.0 to 20.0 mM, 10.0 to 20.0 mM, 0.1 to 10.0 mM, 0.5 to 10.0 mM, 1.0 to 10.0 mM, or 5.0 to 10.0 mM. Alternatively, the amino acid concentration is 1 to 3% by weight. In one specific example, the amino acid is 100 to 200 mM histidine, specifically 150 mM histidine, a pharmaceutically acceptable salt thereof, or a mixture thereof. In one embodiment, the amino acid is 1 to 3% by weight of lysine, arginine, glycine, or proline. The concentration of the amino acid can be adjusted within a range that allows stability of risankizumab or an antigen-binding fragment thereof without affecting the desired pH of the pharmaceutical composition, and may vary depending on the particular amino acid. In one embodiment, the amino acid is 2.5% by weight of proline.

[0022] The polyol not contained in the pharmaceutical composition is specifically sorbitol, sucrose, trehalose, mannose, maltose, mannitol, or a mixture thereof. The pharmaceutical composition contains a metal salt or amino acid stabilizer without a polyol, thereby exhibiting excellent stability (see Test Example 3, Formulations 5 to 9, 14 to 18, 20, and 22; Test Example 4, Formulations 5 to 8). This was an unexpected effect, given that conventional pharmaceutical formulations containing risankizumab generally contain polyols for stability. Furthermore, polyols may contain monosaccharides as impurities. However, monosaccharides are known to induce denaturation (e.g., aggregation) of risankizumab or have the potential to affect protein efficacy (see Non-Patent Documents 1 and 2). However, the pharmaceutical composition does not contain a polyol, thereby avoiding the potential risk of monosaccharides destabilizing the protein active ingredient.

[0023] Furthermore, the pharmaceutical composition may or may not contain a surfactant, and can have excellent stability whether or not it contains a surfactant (see Test Example 3, Formulations 5 to 9, 20 vs. Formulations 14 to 18, 22; see Test Example 4, Formulations 5 to 8).

[0024] Furthermore, the pharmaceutical composition may or may not contain a buffering agent, and can have excellent stability whether or not it contains a buffering agent (see Test Example 3, Formulations 5 to 9, 20 vs. Formulations 14 to 18, 22; see Test Example 4, Formulations 5 to 8).

[0025] In one embodiment, the pharmaceutical composition is free of polyols and surfactants. The pharmaceutical composition provided by the present invention "does not contain component A" means that the pharmaceutical preparation does not contain or is substantially free of component A. "Substantially free of component A" is interpreted to include cases where component A is not contained at all, or where component A, if present, is present in a trace amount that does not substantially affect the properties of the pharmaceutical composition, or where component A is present in an undetectable amount.

[0026] The surfactant may be, for example, a polysorbate, a poloxamer, a sorbitan ester of another fatty acid, or a mixture thereof. The polysorbate may be, for example, polysorbate 20, polysorbate 80, or a mixture thereof. In one embodiment, the surfactant is polysorbate 20. The concentration of the surfactant in the pharmaceutical composition is 0.001 to 2% by weight of the pharmaceutical composition and may vary depending on the specific surfactant. In one embodiment, the surfactant is 0.02% by weight of polysorbate 20.

[0027] The pH of the pharmaceutical composition is 5.0 to 7.0. Specifically, for example, pH 5 to 6.8, pH 5 to 6.5, pH 5 to 6.3, pH 5.2 to 6.3, pH 5.0 to 6.0, pH 5.2 to 6.0, pH 5.0 to 5.8, pH 5.2 to 5.8, pH 5.0 to 5.6, pH 5.2 to 5.6, pH 5.1, pH 5.2, pH 5.3, pH 5.4, pH 5.5, pH 5.7, or pH 6.0. In one specific example, the pH of the pharmaceutical composition is 5.7.

[0028] The pharmaceutical composition may contain a buffer. Examples of the buffer include, but are not limited to, acetate, succinate, citrate, glutamate, glycine, lactate, malate, phosphate, tartrate, or histidine buffers, or any combination thereof. The buffer is present in the pharmaceutical composition at an appropriate concentration to maintain the selected pH during storage. The buffer is present in the pharmaceutical composition at, for example, 10 to 60 mM, and the concentration varies depending on the specific buffer. In one specific example, the buffer is 16 mM histidine. The main ingredient of the pharmaceutical composition, risankisumab, is an antibody used to treat autoimmune diseases, particularly psoriasis or psoriatic arthritis, and is currently marketed under the name SKYRIZI®. It is known that risankizumab can be used to treat autoimmune diseases, inflammatory diseases, respiratory diseases, metabolic disorders, or cancer, more specifically, psoriasis, psoriatic arthritis, inflammatory bowel disease, multiple sclerosis, rheumatoid arthritis, Crohn's disease, ulcerative colitis, ankylosing spondylitis, asthma, or chronic obstructive pulmonary disease (COPD), etc. Risankizumab may also include "biosimilars" or "biobetters" of the active risankizumab present in the commercially available drug SKYRIZI®.

[0029] Risankizumab can be an antibody comprising a light chain having the amino acid sequence of SEQ ID NO:1 and a heavy chain having the amino acid sequence of SEQ ID NO:2.

[0030] In the present invention, the term "antigen-binding fragment" refers to a fragment capable of binding to an antibody, i.e., the target antigen IL-23p19 of risankizumab, and includes, but is not limited to, for example, a Fab fragment, a F(ab')2 fragment, an Fc fragment, or an scFv fragment.

[0031] Risankizumab can be produced by common methods well known in the art. For example, WO2012 / 061448 describes methods that a person skilled in the art can use to produce risankizumab. These methods are incorporated herein by reference.

[0032] The concentration of risankizumab or an antigen-binding fragment thereof in the pharmaceutical composition is, for example, 9 to 170 mg / mL, 9 to 45 mg / mL, 10 to 40 mg / mL, 15 to 35 mg / mL, 20 to 30 mg / mL, 130 to 160 mg / mL, or about 150 mg / mL.

[0033] The pharmaceutical composition may be an aqueous liquid formulation.

[0034] The pharmaceutical composition is administered by parenteral delivery. Parenteral administration includes, for example, subcutaneous, intramuscular, intradermal, and intramedullary injection, as well as intrathecal, direct intraventricular, intravenous, intraperitoneal, and intravitreal injection. Drugs can be administered in a variety of conventional ways, such as intraperitoneal, parenteral, intraarterial, or intravenous injection.

[0035] The pharmaceutical composition is for subcutaneous injection, intramuscular injection, or intravenous injection. The pharmaceutical composition may further contain a suitable aqueous carrier to make it suitable for injection. The aqueous carrier is a pharmaceutically acceptable solution that is safe, non-toxic, and suitable for human administration, such as water, saline solution, infusion solution, dextrose, or a mixture thereof. In one embodiment, the aqueous carrier is water.

[0036] The pharmaceutical composition has an osmolality within a suitable range for subcutaneous or intravenous injection, such as 200 to 400 mOsm / kg, 200 to 350 mOsm / kg, 250 to 300 mOsm / kg, 250 to 290 mOsm / kg, 270 to 328 mOsm / kg, 250 to 269 mOsm / kg, or 328 to 350 mOsm / kg. The osmolality is appropriately adjusted to minimize pain during administration.

[0037] In one specific example, the pharmaceutical composition may have a concentration of the risankizumab or antigen-binding fragment thereof of 9 to 170 mg / mL, the stabilizer is sodium chloride, lysine, arginine, glycine, proline, histidine, or a pharmaceutically acceptable salt thereof, or a mixture thereof, or sodium chloride, the surfactant is polysorbate 20, the pH is 5.0 to 7.0, and the composition may be polyol-free.

[0038] In one specific example, the pharmaceutical composition has a concentration of the risankizumab or antigen-binding fragment thereof of 9 to 170 mg / mL, the stabilizer is sodium chloride, lysine, arginine, glycine, proline, histidine, or a pharmaceutically acceptable salt thereof, or a mixture thereof, or sodium chloride, and has a pH of 5.0 to 7.0, and is free of polyols and surfactants.

[0039] In one embodiment, the pharmaceutical composition comprises 150 mg / mL risankizumab, 16 mM histidine, 2.5% by weight proline, 200.02% by weight polysorbate, and has a pH of 5.7, optionally free of polyols and / or surfactants.

[0040] In one embodiment, the pharmaceutical composition comprises 150 mg / mL risankizumab, 10 mM histidine, 2.5% by weight proline, 200.02% by weight polysorbate, and has a pH of 5.7, optionally free of polyols and / or surfactants.

[0041] In the pharmaceutical composition of the present invention, risankizumab or its antigen-binding fragment is stabilized. The term "stabilization" means that risankizumab or its antigen-binding fragment substantially retains its physical stability, chemical stability, and / or biological activity before and after administration, during additional manufacturing processes, storage, or preservation. The physical stability, chemical stability, and / or biological activity can be evaluated by commonly known methods. In one specific example, the stability can be evaluated by thermal stability, light stability, cold-thaw stability, and agitation stability tests, as described in the following embodiments.

[0042] Risankizumab can be used to treat any disease known in the art to be effective. For example, WO2012 / 061448 lists indications that can be treated by administering risankizumab. These methods are incorporated herein by reference.

[0043] Thus, yet another aspect of the present invention provides a method for treating an autoimmune disease, specifically psoriasis or psoriatic arthritis, comprising administering the pharmaceutical composition to an individual. In addition to the above diseases, other conditions that can be treated include inflammatory diseases, respiratory diseases, metabolic disorders, and cancer, more specifically, inflammatory bowel disease (Crohn's disease, ulcerative colitis), multiple sclerosis, rheumatoid arthritis, ankylosing spondylitis, asthma, and chronic obstructive pulmonary disease (COPD). The method for treating includes administering a therapeutically effective amount of the pharmaceutical preparation to an individual. The individual includes humans. Yet another aspect of the present invention provides the pharmaceutical composition as described above for the treatment of autoimmune diseases, cancer, psoriasis, psoriatic arthritis, inflammatory bowel disease, multiple sclerosis, rheumatoid arthritis, ankylosing spondylitis, asthma, or chronic obstructive pulmonary disease (COPD), etc.

[0044] Yet another aspect of the present invention includes the steps of: adding a stabilizer to an aqueous carrier to prepare a mixed solution; and adding risankizumab or an antigen-binding fragment thereof to the mixed solution; or adding risankizumab or an antigen-binding fragment thereof to an aqueous carrier to produce a solution; and adding a stabilizer to the solution. As a method for preparing the pharmaceutical composition, the preparation step is optionally carried out without adding a polyol.

[0045] The aqueous carrier is an aqueous solvent (eg, water or saline).

[0046] In the method, a buffering agent (or a buffer solution) and / or a surfactant may be optionally added in addition to the step of adding a stabilizer.

[0047] The details of the method for producing the pharmaceutical composition are the same as those described for the pharmaceutical composition according to one aspect of the present invention.

[0048] The configuration and effects of the present invention will be described in more detail below with reference to embodiments and experimental examples. However, the following embodiments and experimental examples are provided for illustrative purposes only to facilitate understanding of the present invention, and the scope and spirit of the present invention are not limited thereby.

[0049] [Table A]

[0050] Analysis method In the analytical methods of the following test examples, SE-HPLC and WCX were performed on the samples to confirm the heat stability, cold-thaw stability, and stirring stability. HMW%, LMW%, and Monomer% were measured through SE-HPLC analysis. Acidic%, Basic%, and Main% were measured through WCX analysis.

[0051] In addition, to confirm the photostability, HIC was performed in addition to SE-HPLC and WCX. Hydrophobic%, hydrohilic%, and main% were measured through HIC analysis.

[0052] The SE-HPLC analytical method uses a high-performance liquid chromatography (HPLC) system, such as a Waters (Milford, MA, USA) Alliance HPLC system containing a size-exclusion chromatography (SEC) column. Proteins separated from the SEC column are detected by UV absorbance at 280 nm, and relative amounts are determined by calculating the area under the curve (AUC) for each separated peak. Peaks are assigned to various species based on their separation times, which correspond to the molecular size of the species. To measure the relative HMW%, LMW%, and Monomer% of antibodies, particularly monomeric antibodies, in the preparation, HMW and LMW, if present in the preparation, are separated from each other. Specifically, the relative content or amount is expressed as a percentage, and the sum of Monomer%, HMW%, and LMW% equals 100%.

[0053] Weak cation exchange chromatography (WCX) uses a high-performance liquid chromatography (HPLC) system, such as a Waters (Milford, MA, USA) Alliance HPLC system containing a WCX column. Proteins separated from the WCX column are detected by UV absorbance at 280 nm, and relative amounts are determined by calculating the area under the curve (AUC) for each resolved peak or group of resolved peaks. Peaks are assigned to various species by their separation time, which corresponds to the surface charge of the antibody species. For stability analysis, the measurements are performed after formulation dispensing (TO) and then after the indicated storage time under the described storage conditions. Acidic% or Basic% includes all peaks before or after the peak of the Main% original antibody variant. These peaks include antibody variants that are more acidic and / or basic than the Main% original antibody variant, or that have more negative or positive charges on the surface under chromatographic conditions. In particular, the relative content or amount is expressed as a percentage value, and the sum of Main%, Acidic% and Basic% is 100%.

[0054] HIC uses a high-performance liquid chromatography (HPLC) system, such as the Alliance HPLC system from Waters (Milford, MA, USA), which includes an HIC column. Proteins separated from the HIC column are detected by UV absorbance at 280 nm, and relative amounts are determined by calculating the area under the curve (AUC) for each separated peak or group of separated peaks. Peaks are assigned to various species based on their separation time, which corresponds to the hydrophobicity of the species. To determine the relative hydrophobic%, hydrohilic%, and main% of the antibody in the preparation, hydrophobic and hydrohilic forms, if present in the preparation, are separated from each other. Specifically, the relative content or amount is expressed as a percentage, and the sum of hydrophobic%, hydrohilic%, and main% equals 100%.

[0055] Test Example 1: Analysis of formulation stability depending on pH and buffer solution Aqueous risankizumab liquid formulations having the compositions shown in Table 1 below were prepared and analyzed for thermal stability, light stability, cold-thaw stability, and stirring stability.

[0056] [Table 1]

[0057] Sample Preparation: To confirm the stability of formulation samples, samples were collected after storage under various stress conditions. Dialysis was performed using each manufactured buffer solution to produce formulations with the target protein concentration. The manufactured formulations were sterile filtered, filled into syringes at 1 mL, and exposed to various stress conditions. The stress conditions were as follows: For thermal stability, samples were stored and collected at 40±2°C for 1, 2, 4, and 5 weeks. For light stability, samples were stored and collected under integrated near-UV light conditions with an illumination intensity of 1.2 million lux-hours or more and 200 watt-hours / cubic meter or more. For cold-thaw stability, samples were collected after five cycles of cold-thawing at room temperature and at or below -60°C. For stirring stability, samples were collected after stirring at 0 rpm and 300 rpm.

[0058] Results: The results of measuring the thermal stability, light stability, cold-thaw stability, and stirring stability are shown in Tables 2 to 28 below.

[0059] [Table 2]

[0060] [Table 3]

[0061] [Table 4]

[0062] The results confirmed by SE-HPLC analysis at a storage temperature of 40°C are shown in Tables 2 to 4.

[0063] During the 5-week storage period, the HMW% increased by 2 to 5%, with the largest increases measured at +4.10%, +4.51%, +4.33%, and +4.99% for formulations 3, 6, 9, and 17, respectively, and the remaining formulations, excluding the formulation that showed the largest increase, measured an average of +2.17%.

[0064] During the 5-week storage period, the LMW% increased by 2 to 14%, with the largest increases measured at +8.60%, +14.25%, +9.50%, and +4.15% for formulations 4, 6, 9, and 17, respectively, and the remaining formulations, excluding the formulation that showed the largest increase, measured an average of +2.35%.

[0065] During the 5-week storage period, the Monomer% decreased by 3 to 19%. The maximum decreases were measured for Formulations 3, 6, 9, and 17 at -12.59%, -18.77%, -13.85%, and -9.14%, respectively, and the remaining formulations, excluding the formulation with the maximum decrease, showed an average decrease of -4.52%.

[0066] [Table 5]

[0067] [Table 6]

[0068] [Table 7]

[0069] The results confirmed by WCX analysis at a storage temperature of 40°C are shown in Tables 5-7.

[0070] During the 5-week storage period, the Acid % increased from 0 to 36%. The maximum increases were measured for Formulations 3, 6, 9, and 9 at +0.14%, +5.75%, and +4.76%, respectively, and the remaining formulations, excluding the formulation that showed the smallest increase, measured an average of +24.78%.

[0071] During the 5-week storage period, Basic% increased by 1 to 56%. The maximum increases were measured for Formulations 3, 6, 9, and 17 at +48.50%, +55.72%, +54.58%, and +15.45%, respectively, with the remaining formulations measuring an average of +8.26%.

[0072] During the 5-week storage period, the Main% decreased by 24 to 62%. The maximum decreases were measured for Formulations 3, 6, 9, and 17 at -48.65%, -61.47%, -59.35%, and -51.71%, respectively, and the remaining formulations, excluding the formulation with the maximum decrease, were measured at an average of -36.96%.

[0073] [Table 8]

[0074] [Table 9]

[0075] [Table 10]

[0076] The results confirmed by SE-HPLC analysis under conditions of illuminance of 1.2 million lux hours or more and integrated near-UV light of 200 watt-hours / cubic meter or more are shown in Tables 8 to 10.

[0077] After light exposure, HMW% increased by 1 to 14%, with the largest increases measured at +11.38% and +13.67% for formulations 16 and 17, and the remaining formulations, excluding the formulation with the largest increase, measured an average of +4.08%.

[0078] After light exposure, there was no significant increase in LMW%, measured at an average of +0.29%.

[0079] After light exposure, the Monomer% increased by 2 to 14%. The largest decreases were measured for Formulations 16 and 17 at -11.60% and -13.82%, respectively, with the remaining formulations excluding the formulation with the largest decrease measuring an average of -4.38%.

[0080] [Table 11]

[0081] [Table 12]

[0082] [Table 13]

[0083] The results confirmed by WCX analysis under integrated near-UV conditions of 1.2 million lux-hours or greater and 200 watt-hours per cubic meter or greater are shown in Tables 11 through 13.

[0084] After light exposure, there was no significant increase in Acidic%. The average was measured at +4.96%.

[0085] After light exposure, there was no significant increase in Basic%, measuring an average of +16.81%.

[0086] After light exposure, there was no significant decrease in Main%. The average was measured at -21.77%.

[0087] [Table 14]

[0088] [Table 15]

[0089] [Table 16]

[0090] The results of the HIC analysis under conditions of illuminance of 1.2 million lux hours or greater and integrated near-UV radiation of 200 watt-hours per cubic meter or greater are shown in Tables 14 through 16.

[0091] After light exposure, the Hydrophobic% increased by 8 to 27%. The smallest increases were measured for Formulations 13, 14, 15, 16, and 17 at +12.69%, +9.63%, +12.79%, +11.36%, and +8.25%, respectively, with the remaining formulations measuring an average of +22.21%.

[0092] After light exposure, there was no significant increase in Hydrophilic%. An average of +2.96% was measured.

[0093] After light exposure, the Main% decreased by 15 to 28%. The smallest decreases were measured for Formulations 13, 14, 15, 16, and 17 as -14.73%, -12.52%, -17.18%, -18.48%, and -14.83%, respectively, and the remaining formulations, excluding the formulation that showed the smallest decrease, were measured at an average of -24.43%.

[0094] [Table 17]

[0095] [Table 18]

[0096] [Table 19]

[0097] After five cycles of freezing and thawing under room temperature and -60°C or lower, the results confirmed by SE-HPLC analysis are shown in Tables 17 to 19.

[0098] After five freeze-thaw cycles, there was no significant increase in HMW%. The average was measured at -0.23%.

[0099] After five freeze-thaw cycles, there was no significant increase in LMW%, with an average measured value of -0.36%.

[0100] After five freeze-thaw cycles, there was no significant increase in Monomer%. The average was measured at +0.59%.

[0101] [Table 20]

[0102] [Table 21]

[0103] [Table 22]

[0104] The results confirmed by WCX analysis after five cycles of freezing and thawing under room temperature and -60°C or lower are shown in Tables 20 to 22.

[0105] After five freeze-thaw cycles, there was no significant increase in Acidic%. The average was measured at +0.71%.

[0106] After five freeze-thaw cycles, there was no significant increase in Basic%, with an average measured value of -0.72%.

[0107] After five freeze-thaw cycles, there was no significant decrease in Main%. The average was measured as +0.01%.

[0108] [Table 23]

[0109] [Table 24]

[0110] [Table 25]

[0111] After stirring at 300 rpm, the results confirmed by SE-HPLC analysis are shown in Tables 23 to 25.

[0112] After stirring, there was no significant increase in HMW%. An average of +0.09% was measured.

[0113] After stirring, there was no significant increase in LMW%, which was measured at an average of -0.79%.

[0114] After stirring, there was no significant decrease in Monomer%, which was measured as an average of +0.70%.

[0115] [Table 26]

[0116] [Table 27]

[0117] [Table 28]

[0118] After stirring at 300 rpm, the results confirmed by WCX analysis are shown in Tables 26 to 28.

[0119] After stirring, there was no significant increase in Acidic%. The average was measured as +0.04%.

[0120] After stirring, there was no significant increase in Basic%, measured at an average of -0.32%.

[0121] After stirring, there was no significant decrease in Main%. The average was measured as +0.30%.

[0122] Discussion of Results: According to the results in Tables 2 to 28, formulations containing various buffers at pH 5.0 to 7.0 were confirmed to be stable under various stress conditions (heat, light, cold-thawing, and agitation stress).

[0123] Test Example 2-1: Analysis of formulation stability with surfactants Aqueous risankizumab liquid formulations having the compositions shown in Table 29 below were prepared and analyzed for thermal stability, light stability, cold-thaw stability, and stirring stability.

[0124] [Table 29]

[0125] Sample Preparation: To confirm the stability of formulation samples, samples were collected after storage under various stress conditions. Dialysis was performed using each manufactured buffer solution to produce formulations with the target protein concentration. The manufactured formulations were sterile filtered, filled into syringes at 1 mL, and exposed to various stress conditions. The stress conditions were as follows: For thermal stability, samples were stored and collected at 40±2°C for 1, 2, 4, and 6 weeks. For light stability, samples were stored and collected under integrated near-UV light conditions with an illumination intensity of 1.2 million lux-hours or more and 200 watt-hours / cubic meter or more. For cold-thaw stability, samples were collected after five cycles of cold-thawing at room temperature and at or below -60°C. For stirring stability, samples were collected after stirring at 0 rpm and 300 rpm.

[0126] Results: The results of measuring heat stability, light stability, stirring stability, and cold-thaw stability are shown in Tables 30 to 56 below.

[0127] [Table 30]

[0128] [Table 31]

[0129] [Table 32]

[0130] The results confirmed by SE-HPLC analysis at a storage temperature of 40°C are shown in Tables 30-32.

[0131] There was no significant increase in HMW% during the 6-week storage period, measured at an average of +2.26%.

[0132] There was no significant increase in LMW% over the 6-week storage period, measured at an average of +2.04%.

[0133] There was no significant decrease in Monomer% during the 6-week storage period, measured at an average of -4.30%.

[0134] [Table 33]

[0135] [Table 34]

[0136] [Table 35]

[0137] The results confirmed by WCX analysis at a storage temperature of 40°C are shown in Tables 33 through 35. There was no significant increase in Acidic% over the 6-week storage period, measured at an average of +25.59%. There was no significant increase in Basic% over the 6-week storage period, measured at an average of +1.76%. There was no significant decrease in Main% over the 6-week storage period, measured at an average of -27.35%.

[0138] [Table 36]

[0139] [Table 37]

[0140] [Table 38]

[0141] The results confirmed by SE-HPLC analysis under conditions of illuminance of 1.2 million lux hours or more and integrated near-UV light of 200 watt-hours / cubic meter or more are shown in Tables 36-38.

[0142] After light exposure, there was no significant increase in HMW%. An average of +6.65% was measured.

[0143] After light exposure, there was no significant increase in LMW%, measured at an average of +0.00%.

[0144] After light exposure, there was no significant decrease in Monomer%. The average was measured at -6.65%.

[0145] [Table 39]

[0146] [Table 40]

[0147] [Table 41]

[0148] The results confirmed by WCX analysis under integrated near-UV conditions of 1.2 million lux-hours or greater and 200 watt-hours / cubic meter or greater are shown in Tables 39 through 41.

[0149] After light exposure, there was no significant increase in Acidic%. The average was measured at +6.73%.

[0150] After light exposure, there was no significant increase in Basic%, measuring an average of +14.19%.

[0151] After light exposure, there was no significant decrease in Main%. The average was measured at -20.92%.

[0152] [Table 42]

[0153] [Table 43]

[0154] [Table 44]

[0155] The results determined by HIC analysis under conditions of illuminance of 1.2 million lux hours or greater and integrated near-UV radiation of 200 watt-hours per cubic meter or greater are shown in Tables 42 through 44.

[0156] After light exposure, there was no significant increase in Hydrophobic%. The average was measured at +15.46%.

[0157] After light exposure, there was no significant increase in Hydrophilic%. An average of +4.84% was measured.

[0158] After light exposure, there was no significant decrease in Main%. The average was measured at -20.31%.

[0159] [Table 45]

[0160] [Table 46]

[0161] [Table 47]

[0162] After five cycles of freezing and thawing under room temperature and -60°C or lower, the results confirmed by SE-HPLC analysis are shown in Tables 45 to 47.

[0163] After five freeze-thaw cycles, there was no significant increase in HMW%. The average was measured at +0.03%.

[0164] After five freeze-thaw cycles, there was no significant increase in LMW%, with an average measured value of +0.03%.

[0165] After five cycles of freezing and thawing, there was no significant increase in the Monomer%. The average was measured at -0.06%.

[0166] [Table 48]

[0167] [Table 49]

[0168] [Table 50]

[0169] The results confirmed by WCX analysis after five cycles of freezing and thawing under room temperature and -60°C or lower are shown in Tables 48 to 50.

[0170] After five freeze-thaw cycles, there was no significant increase in Acidic%. The average was measured at +0.73%.

[0171] After five freeze-thaw cycles, there was no significant increase in Basic%, with an average measured value of -0.09%.

[0172] After five freeze-thaw cycles, there was no significant decrease in Main%. The average was measured at -0.64%.

[0173] [Table 51]

[0174] [Table 52]

[0175] [Table 53]

[0176] After stirring at 300 rpm, the results confirmed by SE-HPLC analysis are shown in Tables 51 to 53.

[0177] After stirring, there was no significant increase in HMW%. An average of -0.02% was measured.

[0178] After stirring, there was no significant increase in LMW%, measured at an average of -0.14%.

[0179] After stirring, there was no significant decrease in Monomer%, with an average measured value of +0.17%.

[0180] [Table 54]

[0181] [Table 55]

[0182] [Table 56]

[0183] After stirring at 300 rpm, the results confirmed by WCX analysis are shown in Tables 54 to 56.

[0184] After stirring, there was no significant increase in Acidic%. The average was measured at -0.04%.

[0185] After stirring, there was no significant increase in Basic%, measured at an average of +0.02%.

[0186] After stirring, there was no significant decrease in Main%. The average was measured as +0.02%.

[0187] Discussion of Results: The results in Tables 30 to 56 show that both surfactant-containing and surfactant-free formulations are stable under various stress conditions (heat, light, cold-thawing, and agitation stress).

[0188] Test Example 2-2: Analysis of formulation stability with surfactants Aqueous risankizumab liquid formulations having the compositions shown in Table 57 below were prepared and analyzed for thermal stability.

[0189] [Table 57]

[0190] Sample preparation: To confirm the stability of the formulation samples, they were collected after storage under various stress conditions. Dialysis was performed using each prepared buffer solution to produce a formulation with the target protein concentration. The prepared formulation was sterile filtered and filled into a syringe at 1 mL to confirm thermal stability. Thermal stability was confirmed by storing and collecting samples at a temperature of 40±2°C for 1, 2, and 4 weeks.

[0191] Results: The results of the thermal stability measurements are shown in Tables 58 to 63 below.

[0192] [Table 58]

[0193] [Table 59]

[0194] [Table 60]

[0195] The results confirmed by SE-HPLC analysis at a storage temperature of 40°C are shown in Tables 58-60.

[0196] There was no significant increase in HMW% over the 4 week storage period, measured at an average of +1.14%.

[0197] There was no significant increase in LMW% over the 4-week storage period, measured at an average of +1.86%.

[0198] There was no significant decrease in Monomer% during the 4-week storage period, measured at an average of -3.00%.

[0199] [Table 61]

[0200] [Table 62]

[0201] [Table 63]

[0202] The results confirmed by WCX analysis at a storage temperature of 40°C are shown in Tables 61 through 63. There was no significant increase in Acidic% over the 4-week storage period, measured at an average of +13.78%. There was no significant increase in Basic% over the 4-week storage period, measured at an average of +8.02%. There was no significant decrease in Main% over the 4-week storage period, measured at an average of -21.79%.

[0203] Discussion of Results: The results in Tables 58 to 63 above show that both surfactant-containing and surfactant-free formulations are stable under heat temperature stress conditions.

[0204] Test Example 3: Analysis of formulation stability with stabilizers Aqueous risankizumab liquid formulations having the compositions shown in Table 64 below were prepared and analyzed for thermal stability, light stability, cold-thaw stability, and stirring stability.

[0205] [Table 64]

[0206] Sample Preparation: To confirm the stability of formulation samples, samples were collected after storage under various stress conditions. Dialysis was performed using each manufactured buffer solution to produce formulations with the target protein concentration. The manufactured formulations were sterile filtered, filled into syringes at 1 mL, and exposed to various stress conditions. The stress conditions were as follows: For thermal stability, samples were stored and collected at 40±2°C for 2, 4, and 6 weeks. For light stability, samples were stored and collected under integrated near-UV light conditions with an illumination intensity of 1.2 million lux-hours or more and 200 watt-hours / cubic meter or more. For cold-thaw stability, samples were collected after five cycles of cold-thawing at room temperature and at or below -60°C. For stirring stability, samples were collected after stirring at 0 rpm and 300 rpm.

[0207] Results: The stability measurements are shown in Tables 65 to 91 below.

[0208] [Table 65]

[0209] [Table 66]

[0210] [Table 67]

[0211] The results confirmed by SE-HPLC analysis at a storage temperature of 40°C are shown in Tables 65 to 67. Over the 6-week storage period, HMW% increased by 0.8 to 2.0%. For formulations containing polyol or salt and surfactant, an average increase of 1.75% was measured (formulations 1 to 5). For formulations containing amino acid and surfactant, an average increase of 1.27% was measured (formulations 6 to 9, formulation 20). For formulations containing polyol or salt, an average increase of 1.78% was measured (formulations 10 to 14). For formulations containing amino acid, an average increase of 1.26% was measured (formulations 15 to 18, formulation 22).

[0212] There was no significant increase in LMW% during the storage period of the six strains, with an average of +2.8% measured.

[0213] During the 6-week storage period, the Monomer% decreased by 3.74 to 4.97%. For formulations containing polyol or salt and surfactant (formulations 1 to 5), an average decrease of 4.41% was measured. For formulations containing amino acid and surfactant (formulations 6 to 9, formulation 20), an average decrease of 4.19% was measured. For formulations containing polyol or salt, an average increase of 4.40% was measured (formulations 10 to 14). For formulations containing amino acid (formulations 15 to 18, formulation 22), an average decrease of 4.16% was measured.

[0214] [Table 68]

[0215] [Table 69]

[0216] [Table 70]

[0217] The results confirmed by WCX analysis at a storage temperature of 40°C are shown in Tables 68 through 70. There was no significant increase in Acidic% over the 6-week storage period. An average of +17.73% was measured.

[0218] There was no significant increase in Basic% over the 6-week storage period, measured at an average of +6.41%.

[0219] There was no significant decrease in Main% during the 6-week storage period. An average decrease of 24.14% was measured.

[0220] According to the results in Tables 65 to 70, it was confirmed that the formulations containing or not containing an amino acid and optionally a surfactant have better stability under thermal conditions than the formulations containing or not containing a polyol and optionally a surfactant.

[0221] [Table 71]

[0222] [Table 72]

[0223] [Table 73]

[0224] The results confirmed by SE-HPLC analysis under conditions of irradiance of 1.2 million lux hours or more and integrated near-UV light of 200 watt-hours / cubic meter or more are shown in Tables 71-73.

[0225] After light exposure, the HMW% increased by 0.5 to 5.2%. For formulations containing polyol or salt and surfactant (formulations 1 to 5), an average increase of 4.27% was measured. For formulations containing amino acid and surfactant (formulations 6 to 9, formulation 20), an average increase of 3.24% was measured. For formulations containing polyol or salt, an average increase of 4.05% was measured (formulations 10 to 14). For formulations containing amino acid (formulations 15 to 18, formulation 22), an average increase of 2.80% was measured.

[0226] After light exposure, there was no significant increase in LMW%, measured at an average of +0.07%.

[0227] After light exposure, the Monomer% decreased by 0.6 to 5.2%. For formulations containing polyol or salt and surfactant (formulations 1 to 5), an average decrease of 4.39% was measured. For formulations containing amino acid and surfactant (formulations 6 to 9, formulation 20), an average decrease of 3.37% was measured. For formulations containing polyol or salt (formulations 10 to 14), an average decrease of 4.13% was measured. For formulations containing amino acid (formulations 15 to 18, formulation 22), an average decrease of 2.82% was measured.

[0228] [Table 74]

[0229] [Table 75]

[0230] [Table 76]

[0231] The results confirmed by WCX analysis under integrated near-UV conditions of 1.2 million lux-hours or greater and 200 watt-hours per cubic meter or greater are shown in Tables 74 through 76.

[0232] After light exposure, the Acid % increased by 2.4 to 9.9%. For formulations containing polyol or salt and surfactant (formulations 1 to 5), an average increase of 7.48% was measured. For formulations containing amino acid and surfactant (formulations 6 to 9, formulation 20), an average increase of 6.87% was measured. For formulations containing polyol or salt (formulations 10 to 14), an average increase of 6.85% was measured. For formulations containing amino acid (formulations 15 to 18, formulation 22), an average increase of 5.41% was measured.

[0233] After light exposure, Basic% increased by 3.3 to 16.0%. For formulations containing polyol or salt and surfactant (formulations 1 to 5), an average increase of 14.70% was measured. For formulations containing amino acid and surfactant (formulations 6 to 9, formulation 20), an average increase of 12.83% was measured. For formulations containing polyol or salt (formulations 10 to 14), an average increase of 13.96% was measured. For formulations containing amino acid (formulations 15 to 18, formulation 22), an average increase of 10.82% was measured.

[0234] After light exposure, the Main% decreased by 5.7 to 25.0%. For formulations containing polyol or salt and surfactant (formulations 1 to 5), an average decrease of 22.18% was measured. For formulations containing amino acid and surfactant (formulations 6 to 9, formulation 20), an average decrease of 19.71% was measured. For formulations containing polyol or salt (formulations 10 to 14), an average decrease of 20.81% was measured. For formulations containing amino acid (formulations 15 to 18, formulation 22), an average decrease of 16.23% was measured.

[0235] [Table 77]

[0236] [Table 78]

[0237] [Table 79]

[0238] The results of the HIC analysis under conditions of illuminance of 1.2 million lux hours or greater and integrated near-UV radiation of 200 watt-hours per cubic meter or greater are shown in Tables 77 through 79.

[0239] After light exposure, the Hydrophobic% increased by 5 to 17%. The smallest increases were measured for Formulations 15, 18, 20, and 22 at +9.68%, +9.33%, +5.89%, and +4.74%, respectively, with the remaining formulations measuring an average of +12.70%.

[0240] After light exposure, the hydrophilic % increased by 0.1 to 3.0%. For formulations containing polyol or salt and surfactant (formulations 1 to 5), an average increase of 2.28% was measured. For formulations containing amino acid and surfactant (formulations 6 to 9, formulation 20), an average increase of 1.65% was measured. For formulations containing polyol or salt, an average increase of 2.23% was measured (formulations 10 to 14). For formulations containing amino acid (formulations 15 to 18, formulation 22), an average increase of 1.42% was measured.

[0241] After light exposure, the Main% decreased by 5 to 19%. The smallest decreases were measured for Formulations 15, 18, 20, and 22 at -11.09%, -11.34%, -6.08%, and -4.85%, respectively, and the remaining formulations, excluding the formulation that showed the smallest decrease, were measured at an average of -14.76%.

[0242] The results in Tables 71 to 79 confirm that the formulations containing or not containing an amino acid and optionally a surfactant have better photostability than the formulations containing or not containing a polyol and optionally a surfactant.

[0243] [Table 80]

[0244] [Table 81]

[0245] [Table 82]

[0246] The results confirmed by SE-HPLC analysis after five cycles of freezing and thawing under room temperature and -60°C or lower are shown in Tables 80 to 82.

[0247] After five freeze-thaw cycles, there was no significant increase in HMW%. The average was measured at +0.03%.

[0248] After five freeze-thaw cycles, there was no significant increase in LMW%, with an average measured value of +0.32%.

[0249] After five cycles of freezing and thawing, there was no significant decrease in the Monomer%. The average was measured at -0.35%.

[0250] [Table 83]

[0251] [Table 84]

[0252] [Table 85]

[0253] The results confirmed by WCX analysis after five cycles of freezing and thawing under room temperature and below -60°C are shown in Tables 83 to 85.

[0254] After five freeze-thaw cycles, there was no significant increase in the acidity, with the average measured value being -0.96%.

[0255] After five freeze-thaw cycles, there was no significant increase in Basic%, with an average measured value of +0.98%.

[0256] After five freeze-thaw cycles, there was no significant decrease in Main%. The average was measured at -0.01%.

[0257] According to the results in Tables 80 to 85, all the formulations were confirmed to be stable under cold-thaw stress conditions.

[0258] [Table 86]

[0259] [Table 87]

[0260] [Table 88]

[0261] After stirring at 300 rpm, the results confirmed by SE-HPLC analysis are shown in Tables 86 to 88. After stirring, there was no significant increase in HMW%. The average was measured to be +0.05%. After stirring, there was no significant increase in LMW%. The average was measured to be -0.06%. After stirring, there was no significant increase in Monomer%. The average was measured to be 0.00%.

[0262] [Table 89]

[0263] [Table 90]

[0264] [Table 91]

[0265] After stirring at 300 rpm, the results confirmed by WCX analysis are shown in Tables 89 to 91. After stirring, there was no significant increase in Acidic%. The average was measured to be -0.22%. After stirring, there was no significant increase in Basic%. The average was measured to be +0.13%. After stirring, there was no significant increase in Main%. The average was measured to be +0.08%.

[0266] According to the results in Tables 86 to 91, all the formulations were confirmed to be stable under agitation stress conditions.

[0267] Discussion of the results: Formulations containing amino acid stabilizers, with or without surfactants, are stable under a variety of stress conditions (heat, light, and agitation stress). In contrast, formulations containing polyol stabilizers tend to be less stable than those using amino acid stabilizers.

[0268] Test Example 4: Analysis of the stability of formulations containing only stabilizers Aqueous risankizumab liquid formulations having the compositions shown in Table 92 below were prepared and analyzed for thermal stability.

[0269] [Table 92]

[0270] Sample preparation: To confirm the stability of formulation samples containing only stabilizers, without buffers or surfactants, samples were collected after storage under heat stress conditions. Dialysis was performed using buffers containing each stabilizer to produce formulations at the target protein concentration. The manufactured formulations were sterile filtered, filled into tubes in 0.3 mL volumes, and exposed to heat stress conditions. Thermal stability was evaluated by storing and collecting samples at 40±2°C for 1, 2, and 4 weeks.

[0271] Results: The results of the thermal stability measurements are shown in Tables 93 to 95 below.

[0272] [Table 93]

[0273] [Table 94]

[0274] [Table 95]

[0275] The results confirmed by SE-HPLC analysis at a storage temperature of 40°C are shown in Tables 93-95.

[0276] During the 4-week storage period, HMW% increased by 0.9 to 1.7%. For formulations containing only polyols (formulations 1-4), an average increase of 1.67% was measured. For formulations containing only amino acids (formulations 5-8), an average increase of 1.10% was measured. The change in HMW% (ΔHMW%) from the initial value at 1, 2, and 4 weeks is shown in Figure 3.

[0277] There was no significant increase in LMW% over the 4-week storage period, measured at an average of +1.39%.

[0278] During the storage period of the four strains, the increase in HMW% resulted in a decrease in Monomer% of 2.3 to 3.1%. For formulations containing only polyols (formulations 1 to 4), an average decrease of 2.95% was measured. For formulations containing only amino acids (formulations 5 to 8), an average decrease of 2.59% was measured.

[0279] The results in Tables 92 to 95 confirm that the formulations containing amino acids have better photostability than the formulations containing polyols.

[0280] Discussion of Results: The results in Tables 93 to 95 above indicate that formulations containing only amino acid stabilizers are more stable under heat stress conditions than formulations containing only polyol stabilizers.

[0281] Test Example 5: Analysis of stability depending on concentration of formulation containing proline stabilizer Aqueous risankizumab liquid formulations having the compositions shown in Table 96 below were prepared and analyzed for thermal stability, light stability, cold-thaw stability, and agitation stability.

[0282] [Table 96]

[0283] Sample Preparation: To confirm the stability of the formulation samples over an appropriate concentration range, a total of five factors (protein concentration, pH, buffer histidine concentration, stabilizer proline concentration, and surfactant concentration) were designed based on DoE (Design of Experiments), and samples were collected after production and storage. Dialysis was performed using each manufactured buffer solution to produce formulations with the target protein concentration. The manufactured formulations were sterile filtered, filled into syringes at 1 mL, and subjected to various stress conditions. The stress conditions were as follows: For thermal stability, samples were stored and collected at 40±2°C for 4 and 6 weeks. For light stability, samples were stored and collected under integrated near-UV light conditions with an illumination intensity of 1.2 million lux-hours or more and 200 watt-hours / cubic meter or more. For cold-thaw stability, samples were collected after five cycles of cold-thawing at room temperature and at temperatures below -60°C. For stirring stability, samples were collected after stirring at 0 rpm and 400 rpm.

[0284] Analysis method: A predictive model was established considering the main effects, interaction effects, and secondary effects of a total of five factors (protein concentration, pH, histidine concentration, proline concentration, and surfactant concentration), and the thermal stability, light stability, cold-thaw stability, and stirring stability at various concentrations were confirmed.

[0285] Results: The results of measuring heat stability, light stability, cold-thaw stability, and stirring stability are shown in Tables 97 to 123 below.

[0286] [Table 97]

[0287] [Table 98]

[0288] [Table 99]

[0289] [Table 100]

[0290] [Table 101]

[0291] [Table 102]

[0292] [Table 103]

[0293] [Table 104]

[0294] [Table 105]

[0295] [Table 106]

[0296] Table 107

[0297] Table 108

[0298] Table 109

[0299] Table 110

[0300] Table 111

[0301] Table 112

[0302] Table 113

[0303] Table 114

[0304] Table 115

[0305] Table 116

[0306] [Table 117]

[0307] [Table 118] [Table 119]

[0308] [Table 120]

[0309] [Table 121]

[0310] [Table 122]

[0311] [Table 123]

[0312] Discussion of results: A DoE statistical analysis of the thermal stability, cold-thaw stability, stirring stability, and light stability test results confirmed the appropriate concentration ranges for each component of the formulation, including proline.

[0313] The graphs obtained by performing stability modeling using DoE statistical analysis on the thermal stability test results (SE-HPLC and WCX results), stirring stability test results (SE-HPLC and WCX results), light stability test results (SE-HPLC, WCX, and HIC results), and cold-thaw stability test results (SE-HPLC and WCX results) are shown in Figures 4, 5, 6, and 7, respectively.

[0314] According to the results shown in Figures 4, 5, 6, and 7, 150 mg / mL risankizumab at pH 5.7, 16 mM histidine, 2.5 wt% proline stabilizer concentration, and 0.02 wt% polysorbate 20 concentration was found to have optimal stability in terms of thermal stability, stirring stability, light stability, and cold-thaw stability.

Claims

1. (a) risankizumab or an antigen-binding fragment thereof; (b) a stabilizer; A polyol-free aqueous pharmaceutical composition.

2. 2. The aqueous pharmaceutical composition according to claim 1, wherein the stabilizer is an amino acid or a pharmaceutically acceptable salt thereof, or sodium chloride.

3. (a) risankizumab or an antigen-binding fragment thereof; (b) an amino acid or a pharmaceutically acceptable salt thereof; (c) An aqueous pharmaceutical composition having a pH of 5.0 to 7.

0.

4. 4. The aqueous pharmaceutical composition of claim 3, which is polyol-free.

5. 4. The aqueous pharmaceutical composition according to claim 2 or 3, wherein the amino acid is lysine, arginine, glycine, proline, histidine, phenylalanine, tyrosine, tryptophan, or a pharmaceutically acceptable salt thereof, or a mixture thereof.

6. 5. The aqueous pharmaceutical composition according to claim 1, wherein the polyol is sorbitol, sucrose, trehalose, mannose, maltose, mannitol, or a mixture thereof.

7. 5. The aqueous pharmaceutical composition according to claim 1, further comprising a surfactant.

8. 5. The aqueous pharmaceutical composition according to claim 1, which does not contain a surfactant.

9. 8. The aqueous pharmaceutical composition of claim 7, wherein the surfactant is a polysorbate, a poloxamer, a sorbitan ester of another fatty acid, or a mixture thereof.

10. 10. The aqueous pharmaceutical composition of claim 9, wherein the polysorbate is polysorbate 20, polysorbate 80, or a mixture thereof.

11. 3. The aqueous pharmaceutical composition according to claim 1, wherein the pH is 5.0 to 7.

0.

12. 5. The aqueous pharmaceutical composition according to claim 1, further comprising a buffering agent.

13. 13. The pharmaceutical composition of claim 12, wherein the buffering agent is acetate, succinate, citrate, glutamate, glycine, lactate, maleate, phosphate, tartrate, histidine, or any combination thereof.

14. 5. The aqueous pharmaceutical composition of claim 1, wherein the concentration of the risankizumab or antigen-binding fragment thereof is 9 to 170 mg / ml.

15. 5. The aqueous pharmaceutical composition according to claim 1, which is for subcutaneous injection, intramuscular injection, or intravenous injection.

16. 5. The aqueous pharmaceutical composition according to claim 1, which is for the treatment of autoimmune diseases, cancer, psoriasis, psoriatic arthritis, inflammatory bowel disease, multiple sclerosis, rheumatoid arthritis, ankylosing spondylitis, asthma, or chronic obstructive pulmonary disease (COPD).

Images