NZ757965A - Liquid formulation of anti-tnf alpha antibody - Google Patents

Abstract

The present invention relates to a liquid formulation of an anti-TNF alpha antibody, particularly adalimumab. Particularly, the present invention relates to a liquid formulation comprising an anti-TNF-α antibody at a concentration of 50 mg/mL to 130 mg/mL, a stabilizer, a surfactant at a concentration of 0.5 mg/mL to 1.5 mg/mL, a methionine at a concentration of 2.5 mM to 35 mM, and arginine at a concentration of 20 mM to 120 mM, wherein the stabilizer is a polyol, an amino acid other than arginine, or a combination thereof, wherein the polyol is present at a concentration of 25 mg/mL to 82.5 mg/mL and wherein the amino acid other than arginine is present at a concentration of 10 mM to 160 mM, wherein the liquid formulation does not comprise a buffer.

Description

The present invention s to a liquid formulation of an anti-TNF alpha antibody, ularly adalimumab. Particularly, the present invention relates to a liquid formulation comprising an anti-TNF-α antibody at a concentration of 50 mg/mL to 130 mg/mL, a stabilizer, a surfactant at a concentration of 0.5 mg/mL to 1.5 mg/mL, a methionine at a concentration of 2.5 mM to 35 mM, and arginine at a concentration of 20 mM to 120 mM, wherein the stabilizer is a polyol, an amino acid other than arginine, or a combination f, n the polyol is present at a concentration of 25 mg/mL to 82.5 mg/mL and wherein the amino acid other than arginine is present at a concentration of 10 mM to 160 mM, wherein the liquid formulation does not comprise a buffer.

NZ 757965 [DESCRIPTION] [Invention Title] LIQUID FORMULATION OF ANTI-TNF ALPHA ANTIBODY [Technical Field] The present invention relates to a liquid formulation of an anti-TNF-α dy, and specifically to a liquid formulation of adalimumab.

[Background Art] Tumor necrosis factor alpha (TNF-α) is a cytokine produced by various cell types such as monocytes, macrophages, etc. by stimulation of endotoxin, etc. TNF-α is a major mediator of major inflammatory, immunological, and pathophysiological reactions that tes TNF receptors and induces responses such as activation of T cells, proliferation of ytes, etc. (Grell, M., et al. (1995) Cell, 83: 793 to 802).

Adalimumab is a recombinant human immunoglobulin G1 monoclonal antibody that inhibits immune responses induced by TNF-α by selectively binding to TNF-α in vivo. umab was developed by BASP Bioresearch Corporation in 1993 and approved through Abbott Laboratories for sale as a treatment for rheumatoid arthritis. Adalimumab is currently sold under the name HUMIRA®, and upon receipt of approval for its sale as a therapeutic agent for rheumatoid arthritis, HUMIRA® has been used for the ent of Crohn’s disease, ankylosing spondylitis, tic arthritis, tive colitis, etc.

Adalimumab is the first fully human antibody developed as a drug. Adalimumab was developed by applying phage display technology and has enhanced affinity by inducing a mutation in CDRs. umab, also called D2E7, consists of 1,330 amino acids and has a molecular weight of about 148 kD (US Patent No. 6090382). Adalimumab is a TNF-α inhibitor that binds to TNF and blocks TNF-induced reactions by preventing TNF from interacting with cell e TNF receptors (i.e., p55 and p75).

Meanwhile, antibody drugs are a kind of protein drug, and physical and chemical denaturation may occur due to s factors. As the protein denaturation, a chemical denaturation such as oxidation, deamidation, and ization or a structural denaturation such as fragmentation or aggregation may occur. When protein is denatured, it is possible that the protein may lose its own pharmacological activity and thereby induce ssary immune responses in vivo as a side effect. When an antibody is fragmented, its binding affinity or retention time in the body is altered, and thereby the cological activity of the antibody is affected. Additionally, studies have shown that fragmented dies can induce aggregation of antibodies. Additionally, pharmacological activity may be reduced by aggregation. According to studies, as a result of comparison of commercial interferon beta products, it was confirmed that those products which have high contents of ates and particles are highly likely to have a high percentage of neutralizing antibodies in the body (Barnard et al., 2013, J. Pharm. Sci. 102: 915). If neutralizing antibodies are produced in vivo, neutralizing antibodies bind to a protein drug when the drug is injected into the body, thereby affecting the safety, pharmacological effect, and pharmacokinetics of the protein drug.

Additionally, the denaturation and aggregation of epoetin alfa were also shown to be the causes of the increased immunogenicity of epoetin alfa drugs. Accordingly, in the case of protein drugs, it is very important to prepare the protein drugs into appropriate formulations so as not to lose their physiological activity during the period of storage, and also that the proteins are not fragmented, aggregated, or formed into particles. ingly, active studies on the formulations for various protein drugs are underway.

The object of the study on protein formulations is to find an optimal combination by appropriately mixing several additives considering the properties of each product so that the product can be stably stored until it is administered to a t. The main object of adding additives is to stabilize proteins and control the physical properties of mixed materials.

Additives can be d into surfactants, stabilizers, preservatives, buffers, isotonic agents, etc. depending on their purposes and characteristics. In the case of antibody drugs, a greater amount of a protein is administered compared to other protein drugs so as to achieve ive eutic s. In case of a subcutaneous injection, it is important to develop a high-concentration formulation due to difficulties of delivering a large volume at one time, such as the patient’s pain, production, etc.,. As the concentration of a protein increases, its intermolecular interaction ses, thus g problems such as increased ation, increased viscosity, gelation, precipitation, etc. Among these, an excessive increase in viscosity makes production difficult, and also makes the administration to a patient difficult due to an increase in the ion pressure. ore, various methods for predicting and lowering the viscosity of a high-concentration antibody solution are being studied.

[Disclosure] [Technical Problem] An object of the present invention is to provide a liquid formulation of an anti-TNF-α antibody. r object of the present invention is to provide a method for preparing the liquid formulation.

Still another object of the present invention is to provide a method for improving stability of an NF-α antibody using a ition ning a stabilizer, a surfactant, and arginine.

Still another object of the present invention is to provide a method for improving stability of an anti-TNF-α antibody using a composition containing a stabilizer, a tant, and arginine without a buffer.

[Technical on] Hereinafter, the t invention will be described in more detail.

Meanwhile, each of the explanations and exemplary embodiments disclosed herein can be applied to other explanations and exemplary embodiments. That is, all of the ations of s factors disclosed herein belong to the scope of the present ion. Furthermore, the scope of the present invention should not be limited by the specific disclosure provided hereinbelow.

Additionally, an ordinary person skilled in the art will be able to recognize or m using no more than routine experimentation with respect to a number of equivalents to the specific embodiments of the invention described in the present invention. Additionally, such equivalents are intended to be included in the present invention.

To achieve the above objects, an aspect of the present invention provides a liquid formulation of an anti-TNF-α antibody. ing to a first aspect, the present invention provides a liquid formulation of an adalimumab comprising an adalimumab at a concentration of 50 mg/mL to 130 mg/mL, a stabilizer, a surfactant at a concentration of 0.5 mg/mL to 1.5 mg/mL, a methionine at a concentration of 2.5 mM to 35 mM, and arginine at a concentration of 20 mM to 120 mM, wherein the stabilizer is a polyol, an amino acid other than arginine, or a combination f, wherein the polyol is present at a concentration of 25 mg/mL to 82.5 mg/mL and wherein the amino acid other than arginine is present at a concentration of 10 mM to 160 mM, wherein the liquid formulation does not comprise buffer.

According to a second aspect, the present invention provides a method for preparing the liquid formulation of the invention, comprising mixing an umab at a concentration of 50 mg/mL to 130 mg/mL, a stabilizer, a surfactant at a concentration of 0.5 mg/mL to 1.5 mg/mL, a methionine at a tration of 2.5 mM to 35 mM, and arginine at a concentration of 20 mM to 120 mM, wherein the stabilizer is a polyol, an amino acid other than arginine, or a combination thereof, n the polyol is present at a tration of 25 mg/mL to 82.5 mg/mL and wherein the amino acid other than arginine is present at a concentration of 10 mM to 160 mM, wherein the liquid formulation does not comprise buffer.

According to a third , the present invention provides a liquid formulation according to the ion, substantially as herein described with reference to any one of the embodiments of the invention illustrated in the accompanying figures and/or examples, or part thereof.

According to a fourth aspect, the present invention provides a method according to the invention, substantially as herein described with reference to any one of the embodiments of the invention illustrated in the accompanying figures and/or examples, or part thereof.

As used herein, the term "anti-TNF-α dy" refers to an antibody that binds to TNF-α and regulates its biological activity. More specifically, the antibody may bind to TNF-α and inhibit the binding between TNF-α and its receptors, thereby inhibiting signaling by TNF-α. In addition, the anti-TNF-α dy may be a monoclonal dy. 3a followed by page 4 The anti-TNF-α antibody may be in the form of a full-length antibody or an antibody fragment containing an antigen-binding region thereof, but the anti-TNF-α antibody is not ularly limited thereto.

More specifically, the anti-TNF-α antibody may be a recombinant human immunoglobulin G1 monoclonal dy, and even more specifically, may be adalimumab.

The information on adalimumab can be easily obtained by those skilled in the art from known databases.

The antibody may be produced through recombinant DNA technology using a mammalian cell expression system, but the preparation method is not limited thereto.

The antibody may be ned in the liquid formulation according to the present invention in a therapeutically effective amount. In a specific ment, the antibody may be present at a tration of 1 mg/mL to 250 mg/mL, specifically 20 mg/mL to 200 mg/mL, more specifically 50 mg/mL to 200 mg/mL, and even more specifically at a concentration of 50 mg/mL, 100 mg/mL, or 130 mg/mL, but the antibody concentration is not particularly limited thereto.

The liquid formulation of the present invention may contain a stabilizer, a surfactant, and arginine, in on to the anti-TNF-α antibody. The liquid formulation may be a solution formulation capable of stably storing the anti-TNF-α antibody.

Specifically, the stability of the anti-TNF-α antibody may be measured using any protein stability assay well known in the art. Stability may be measured at a selected temperature during the selected time. For a rapid test, the formulation may be stored at a higher or "accelerated temperature" (e.g., at 40°C for 2 weeks to 1 month or more), at which point the time-dependent stability is measured.

In the present invention, "providing stability to an anti-TNF-α dy" means that the loss of an active ingredient under n storage ions (specifically a certain ature) for a certain period of time is less than a specified amount (e.g., less than 10%).

Usually, when a residual rate of the anti-TNF-α antibody in the formulation is 90% or more, specifically about 92% or more at 5±3°C for 2 years, at 25±2°C for 6 months, or 40±2°C for 1 to 2 months, these ations can be interpreted as being stable.

The stabilizer to be contained in the liquid formulation of the present invention may be a , an amino acid, or a combination thereof. The amino acid may be an amino acid other than arginine.

Specifically, the stabilizer may be 1) one kind of a polyol; 2) a combination of one kind of a polyol and one kind of an amino acid; 3) a combination of one kind of a polyol, a first amino acid, and a second amino acid; 4) a combination of a first polyol and a second polyol; 5) a ation of a first polyol, a second polyol, and one kind of an amino acid; 6) a ation of a first polyol, a second polyol, a first amino acid, and a second amino acid; or 7) one kind of an amino acid.

More specifically, the polyol may be mannitol, e, trehalose, PEG, or a combination thereof, and more specifically, sucrose, trehalose, PEG, or a combination thereof.

The PEG may specifically be PEG400 or PEG4000, but is not particularly limited thereto.

In the above preparation, the polyol may be present at a concentration of 0.1 mg/mL to 100 mg/mL.

More specifically, the amino acid other than arginine may be glycine, leucine, isoleucine, phenylalanine, or proline. In the above formulation, the amino acid may be present at a concentration of 1 mM to 300 mM.

Additionally, as used herein, the term "amino acid" also includes all of those in the form of analogues, solvates, hydrates, stereoisomers, and pharmaceutically acceptable salts of the corresponding amino acid that exhibit substantially the same effect.

As used herein, the term aceutically acceptable salt" includes pharmaceutically acceptable inorganic acids, organic acids, or salts derived from bases.

Examples of riate acids may include hloric acid, bromic acid, sulfuric acid, nitric acid, perchloric acid, fumaric acid, maleic acid, phosphoric acid, glycolic acid, lactic acid, salicylic acid, succinic acid, toluene-p-sulfonic acid, tartaric acid, acetic acid, citric acid, methanesulfonic acid, formic acid, benzoic acid, c acid, naphthalenesulfonic acid, benzenesulfonic acid, etc. Examples of salts derived from appropriate bases may include an alkali metal such as , potassium, etc.; an alkali earth metal such as magnesium, etc.; um, etc.

Additionally, as used herein, the term "solvate" means that an amino acid or a salt thereof forms a complex with a solvent molecule.

More specifically, the izer may be one selected from the group consisting of (i) sucrose or trehalose, (ii) PEG having a number average molecular weight of 200 to 600 or PEG having a number e molecular weight of 1000 to 8000; (iii) glycine or leucine; and (iv) a combination of at least two among (i) to (iii), but the stabilizer is not particularly limited thereto.

In a more specific embodiment, the stabilizer may be one selected from the group consisting of 1) any one of sucrose, trehalose, and PEG400; 2) a combination of sucrose or trehalose with glycine or leucine; 3) a combination of sucrose or trehalose with glycine and e; 4) a combination of sucrose or trehalose with PEG4000; 5) a combination of sucrose or trehalose with PEG4000 and glycine; 6) a combination of sucrose or trehalose with PEG4000 and leucine; 7) a combination of e or trehalose with PEG4000, glycine, and leucine; and 8) glycine, but the stabilizer is not particularly limited thereto.

The surfactant to be contained in the liquid formulation of the t invention may be a nonionic surfactant. More specifically, the surfactant may be polysorbate or ically, the surfactant may be rbate 80, polysorbate 20, or poloxamer 188, but the surfactant is not particularly limited thereto.

In the above formulation, the surfactant may be present at a concentration of 0.1 mg/mL to 5 mg/mL.

The arginine to be contained in the liquid formulation of the present invention may be present in the form of a salt, and more ically, in the form of a pharmaceutically acceptable salt.

More ically, the arginine may be in the form of arginine hydrochloride, but the arginine is not particularly limited thereto.

In the above formulation, the arginine may be present at a concentration of 0.1 mM to 200 mM. More specifically, the ne may be present at a concentration of 0.1 mM to 140 mM when the antibody is present in the formulation at a tration of 100 mg/mL, whereas the arginine may be present at a concentration of 0.1 mM to 100 mM when the antibody is present in the formulation at a concentration of 50 mg/mL, but the arginine concentration is not particularly limited thereto.

The arginine may be contained as a ity-reducing agent in the liquid formulation of the present invention.

By containing arginine, the liquid formulation of the present invention may have a viscosity of about 1 cps to 6 cps, but the viscosity is not particularly limited thereto. The ity may be ed using various methods known in the art, for example, in the same manner as described in Example 1, but the measurement method is not particularly limited thereto.

The liquid formulation of the present invention may further contain an idant.

As used herein, the "antioxidant" may act to inhibit the generation of ties that may occur by the oxidation reaction of proteins in a solution state.

Examples of the antioxidant may include sodium hydrogen sulfate, ascorbic acid, ascorbyl palmitate, citric acid, butylhydroxyanisole (BHA), butylhydroxytoluene (BHT), thioglycerol, propyl gallate, methionine, sodium ascorbate, sodium citrate, sodium e, sodium sulfite, EDTA, and other antioxidants, but the antioxidant is not limited thereto.

In the above formulation, the idant (specifically methionine) may be present at a tration of 1 mM to 50 mM, but the antioxidant is not particularly limited thereto.

Additionally, the liquid formulation of the present ion may have a pH of 4 to 6, but the pH is not particularly limited thereto.

Meanwhile, the liquid formulation may not further contain a salt and/or a buffer, but the liquid formulation is not particularly limited thereto.

In a non-limiting embodiment, the formulation containing the anti-TNF-α antibody at a tration of 50 mg/mL or higher may not further contain a salt and/or a buffer.

As confirmed in an embodiment of the present ion, the formulation according to the present invention which does not contain a salt and a buffer can provide greater stability of the NF-α antibody to heat, compared to formulations which do not contain a salt, a buffer, or both of these, but the formulation according to the present invention is not particularly limited thereto. ile, if an additional buffer is not added to the formulation of a high-concentration antibody drug (e.g., a ment where antibodies are present at a concentration of 50 mg/mL or higher) or an additional additive is not added to the formulation in such an amount that makes the c re of the solution deviate from the c re range similar to that of body fluids, the pain at the time of administration can be d, thereby improving patient convenience.

Meanwhile, the liquid formulation according to the present invention may have the following effects, but the effects are not particularly limited thereto.

The liquid formulation containing ne according to the present invention may exhibit a relatively low content of high molecular weight (HMW) products due to the inhibition of aggregation of the anti-TNF-α dy proteins compared to a formulation containing no arginine; and/or may include a relatively low amount of acidic variants of antibody due to the inhibition of production of acidic variants of antibody, compared to a formulation containing no arginine. In particular, the liquid formulation ning arginine according to the present invention may have an effect of reducing antibody produced by denaturation and/or effects of reducing aggregation and le formation in response to certain physical stresses.

The solution formulation containing arginine according to the present ion may further contain a preservative. The preservative refers to a compound added to a pharmaceutical ation to act as an antimicrobial agent. Examples of the preservative may include benzalkonium chloride, benzethonium, chlorohexidine, , m-cresol, benzyl alcohol, methyl paraben, propyl paraben, chlorobutanol, o-cresol, p-cresol, chlorocresol, phenylmercuric nitrate, thimerosal, benzoic acid, etc., but the preservative is not limited thereto. These preservatives may be used alone or in any combination of at least two kinds.

The liquid formulation according to the present invention may be in the form of a pharmaceutical composition.

Additionally, the liquid formulation according to the present invention may contain various pharmaceutically acceptable carriers in addition to the ients described above.

The formulations of the present invention may be used for the prevention or treatment of toid arthritis, psoriasis, psoriatic arthritis, axial spondyloarthritis (e.g., ankylosing spondylitis, severe axial spondyloarthritis without radiographic evidence of ankylosing spondylitis), vasculitis, Alzheimer’s disease, ulcerative colitis, Behcet’s enteritis, hidradenitis suppurativa, uveitis, juvenile idiopathic arthritis, pediatric plaque psoriasis, or Crohn’s e (including adult Crohn’s disease and pediatric Crohn’s disease), but the diseases for prevention or ent are not limited thereto.

The formulations according to the present ion may be administered into the body by oral administration or parenteral administration including subcutaneous, intramuscular, intraperitoneal, ternal, percutaneous, and enous injections and infusions, but the administrations are not limited thereto.

Still another aspect of the present ion provides a method for preparing the liquid formulation of an anti-TNF-α antibody, including mixing an anti-TNF-α antibody, a stabilizer, a surfactant, and arginine with one r.

The above terms are the same as explained above. Additionally, it is obvious that all of the specific embodiments of these terms will also apply to the present aspect.

Still another aspect of the present invention provides a method for improving stability of an anti-TNF-α antibody using a composition that contains a stabilizer, a tant, and arginine.

The above terms are the same as ned above. Additionally, it is s that all of the specific embodiments of these terms will also apply to the present aspect.

Another object of the present invention is to provide a method for improving ity of an anti-TNF-α dy using a composition that contains a stabilizer, a surfactant, and arginine without a buffer.

The above terms are the same as explained above. Additionally, it is obvious that all of the specific embodiments of these terms will also apply to the present aspect.

[Advantageous Effects of the Invention] A liquid formulation of the anti-TNF-α antibody according to the present invention, specifically a liquid formulation of adalimumab, can reduce the formation of adalimumab by-products during storage, thus enabling long-term e. Additionally, adalimumab’s pharmacological effect can be stably preserved for a long period of time by preventing denaturation and aggregation caused in response to physical stresses during the processes of tion and transportation. Accordingly, the liquid formulation according to the present invention can effectively be used in the therapeutic field associated with the adalimumab’s pharmacological cy.

[Brief ption of Drawings] shows the viscosity of each sample in Example 1 according to the additive compositions. shows the number of particles in each sample and placebo thereof in Example 7 before and after passing through a pump.

[DETAILED DESCRIPTION OF THE INVENTION] Hereinafter, the present invention will be bed in more detail with reference to the following Examples. However, these Examples are for illustrative purposes only and the invention is not limited by these Examples. Effect of decreasing viscosity of umab solution ing to additives and confirmation of stability To confirm the additives to be used for the preparation of a liquid formulation of adalimumab, Formulation 1 with a composition of e (55 mg/mL), methionine (5 mM), polysorbate 80 (1 mg/mL), and adalimumab (100 mg/mL) at pH 5.2 was prepared.

Additionally, Formulations 2 to 13 were prepared by further adding each of arginine hydrochloride, lysine hydrochloride, leucine, isoleucine, phenylalanine, glutamic acid, e, proline, alanine, sodium chloride, calcium chloride, and magnesium chloride to the composition of Formulation 1, and the viscosity of each formulation was measured using the mVROC apparatus (Rheosense Inc.). The kinds and trations of the additives added to the formulations and the viscosity of each formulation are shown in Table 1 below and [Table 1] Formulation No. Composition of Additives ity (cp) Formulation 1 - 3.23 Formulation 2 Arginine hydrochloride (20 mM) 3.04 Formulation 3 Lysine hydrochloride (40 mM) 3.14 Formulation 4 Leucine (40 mM) 3.28 Formulation 5 Isoleucine (40 mM) 3.26 Formulation 6 alanine (15 mM) 3.21 ation 7 Glutamic acid (7.5 mM) 3.07 Formulation 8 Glycine (40 mM) 3.20 Formulation 9 Proline (40 mM) 3.20 Formulation 10 Alanine (40 mM) 3.21 Formulation 11 Sodium chloride (40 mM) 3.12 Formulation 12 Calcium chloride (20 mM) 2.94 Formulation 13 Magnesium chloride (20 mM) 3.01 Referring to the viscosity for each formulation shown in Table 1, the viscosity for Formulation 1 consisting of sucrose, methionine, polysorbate 80, and adalimumab was 3.23.

In comparison, the viscosities of the formulations containing amino acids such as leucine, isoleucine, phenylalanine, glycine, and proline were in the range of 3.20 to 3.28, thus not showing any significant difference from that of Formulation 1. In contrast, in cases where ne hydrochloride, lysine hydrochloride, ic acid, sodium de, calcium chloride, magnesium chloride, etc. were added, it was confirmed that the viscosities were in the range of 2.94 to 3.14, thus showing a decrease compared to that of Formulation 1.

To compare the stability of each formulation, each formulation was ized by filtration with a filter (pore size: 0.2 μm), sed into glass syringes (volume: about 1.0 mL) in an amount of 0.4 mL each, stoppered, and stored at 40°C for 2 months. After storage, each sample was analyzed by SE-HPLC to analyze the contents of high molecular weight ties (HMW) (e.g., oligomers, aggregates, etc.) and low molecular weight impurities (LMW) (i.e., nts of adalimumab molecules). First, the SE-HPLC results of formulations with reduced viscosity (Formulations 2, 3, 7, 11, 12, and 13) and that of Formulation 1 as the l group are shown in Table 2 below. Additionally, the SE-HPLC results of formulations with no significant changes in viscosity (Formulations 4, 5, 6, 8, 9, and 10) and that of Formulation 1 as the control group are shown in Table 3 below.

[Table 2] Before Storage After Storage at 40°C for 2 Months HMW (%) LMW (%) Total (%) HMW (%) LMW (%) Total (%) Formulation 1 0.33 0.54 0.87 1.46 5.13 6.59 Formulation 2 0.33 0.53 0.86 1.31 5.21 6.52 Formulation 3 0.33 0.53 0.85 1.42 5.18 6.61 Formulation 7 0.33 0.56 0.89 1.40 5.06 6.45 Formulation 11 0.35 0.55 0.89 1.70 5.40 7.10 ation 12 0.33 0.54 0.87 1.36 6.02 7.38 ation 13 0.34 0.54 0.88 1.72 5.70 7.42 [Table 3] Before Storage After Storage at 40°C for 2 Months HMW (%) LMW (%) Total (%) HMW (%) LMW (%) Total (%) Formulation 1 0.33 0.54 0.87 1.46 5.13 6.59 Formulation 4 0.33 0.52 0.85 1.27 4.81 6.07 Formulation 5 0.33 0.56 0.88 1.28 4.74 6.02 ation 6 0.34 0.52 0.86 1.32 4.85 6.17 Formulation 8 0.33 0.48 0.81 1.21 4.77 5.98 Formulation 9 0.32 0.55 0.87 1.14 4.80 5.94 Formulation 10 0.34 0.56 0.90 1.24 4.84 6.08 Referring to Table 2 above, the HMW, LMW, and total amounts of impurities before storage were shown to be similar among the ations. However, the total amount of impurities, after the storage at 40°C for 2 months, was 7.10% in Formulation 11 containing sodium chloride, 7.38% in Formulation 12 containing calcium chloride, and 7.42% in Formulation 13 containing magnesium chloride, thus showing a significant increase compared to 6.59% of Formulation 1 (i.e., the control group), respectively. In contrast, in the cases of Formulations 2, 3, and 7, where arginine hydrochloride, lysine hloride, and glutamic acid were contained, respectively, the total amounts of impurities after the storage at 40°C for 2 months were shown to be in the range of 6.45% to 6.61%, thus maintaining levels of the total amount of impurities similar to 6.59% of Formulation 1 (i.e., the control group).

Referring to Table 3 above, in the cases of the formulations where amino acids such as leucine, isoleucine, phenylalanine, glycine, e, etc. were added, the total amounts of impurities after the storage at 40°C for 2 months were shown to be in the range of 5.94% to 6.17%, thus showing a significant se compared to 6.59% of Formulation 1 (i.e., the control group). From these results, it was confirmed that amino acids, such as leucine, cine, phenylalanine, e, proline, etc. can contribute to the stability of adalimumab. Stability evaluation 1 according to arginine hydrochloride content To evaluate the stability of adalimumab formulations according to the content of arginine hydrochloride, formulations were prepared as follows.

Formulation 14 was prepared to contain sucrose (55 mg/mL), methionine (5 mM), polysorbate 80 (1 mg/mL), and adalimumab (100 . Formulations 15 and 16 were prepared by adding arginine hydrochloride (20 mM) and arginine hydrochloride (40 mM) to the composition of Formulation 14 and filled into 1mL glass syringes in an amount of 0.4 mL, respectively. Each syringe was stored at 40°C for 2 months and then subjected to SE-HPLC analysis for stability analysis. The composition of each formulation and HMW contained therein before and after storage are shown in Table 4 below.

[Table 4] HMW (%) Formulation Composition Before Storage at After Storage at 40°C for 2 Months 40°C for 2 Months Sucrose (55 mg/mL), Methionine (5 mM), Polysorbate 80 ation 14 0.17 1.00 (1 mg/mL), and Arginine hydrochloride (0 mM) Formulation 15 e (55 , Methionine 0.17 0.82 (5 mM), Polysorbate 80 (1 mg/mL), and Arginine hydrochloride (20 mM) Sucrose (55 mg/mL), Methionine (5 mM), Polysorbate 80 ation 16 0.16 0.82 (1 mg/mL), and Arginine hydrochloride (40 mM) Referring to the SE-HPLC results in Table 4, when the content of arginine hydrochloride was increased from 0 mM (Formulation 14) to 20 mM (Formulation 15) and 40 mM (Formulation 16), the amount of HMW of the samples before storage was r, in the range of 0.16% to 0.17%. However, the content of HMW after the storage at 40°C for 2 months was 1.00% in Formulation 14 (containing no ne hydrochloride), and 0.82% in Formulations 15 and 16 ining 20 mM arginine hydrochloride and 40 mM arginine hydrochloride, respectively), thus showing that the formulations containing 20 mM arginine hydrochloride and 40 mM arginine hydrochloride had reduced levels of HMW compared to the formulation containing no ne hydrochloride. Accordingly, it was confirmed that arginine hydrochloride has a preventive effect against aggregation of adalimumab. Comparison of formation of charge variants of antibody according to ne To compare the formation levels of charge variants of antibody according to arginine contents, a formulation containing arginine and a formulation ning no ne were prepared, stored at 40°C for 1 month, and the features of producing charge variants were compared by CEX-HPLC. Each of the formulations and the contents of charge variants before and after storage are shown in Table 5 below.

[Table 5] For Summary of Before Storage at 40°C for 1 After Storage at 40°C for 1 mula Formulations Month Month tion Acidic K0 K1 Basic Acidic K0 K1 Basic e (55 mg/mL), Methionine (5 mM), Polysorbate 80 A 13.36 70.77 12.44 3.44 33.39 51.58 10.70 4.33 (1 mg/mL), and Adalimumab (100 mg/mL) Sucrose (55 mg/mL), Methionine (5 mM) rbate 80 (1 mg/mL), B 13.15 71.14 12.41 3.30 31.31 52.51 11.16 5.01 (100 mg/mL), and Arginine hydrochloride (20 mM) Formulation A was ed to contain sucrose (55 mg/mL), methionine (5 mM), polysorbate 80 (PS80: 1 mg/mL), and adalimumab (100 mg/mL), and Formulation B was prepared to further contain arginine hydrochloride (20 mM) in the composition of Formulation A. The contents of acidic variants of these two formulations before the storage at 40°C for 1 month were similar. Upon comparison of these two formulations after the storage, it was confirmed that Formulation B ning arginine hydrochloride had a lower content of acidic variants of antibody and a higher content of K0 compared to Formulation A.

Accordingly, it was confirmed that arginine hydrochloride lowers the formation of acidic variants of adalimumab. Formulations according to surfactant types and comparison of stability of ations thereof To compare the ity of liquid formulations of adalimumab according to surfactant types, formulations having the following compositions were prepared.

As in ation 14 of Example 2, Formulation 17 was prepared to contain sucrose (55 mg/mL), methionine (5 mM), polysorbate 80 (1 mg/mL), and adalimumab (100 mg/mL).

Additionally, Formulations 18 and 19 were prepared by varying the tant type to polysorbate 20 and poloxamer 188 while fixing the amount of the tant to be the same.

Each formulation was sterilized by filtration, filled into each 1 mL glass syringe in an amount of 0.4 mL, and stored at 40°C for 1 month. The amounts of impurities of HMW and LMW in samples before and after the storage were analyzed by C. The compositions of Formulations 17 to 19 and the SE-HPLC contents of these formulations before and after the storage at 40°C for 1 month were analyzed and the results are shown in Table 6 below.

[Table 6] Before Storage at After Storage at 40°C 40°C for 1 Month for 1 Month HMW LMW Total HMW LMW Total (%) (%) (%) (%) (%) (%) Formulation 17 Sucrose (55mg/mL), 0.14 0.37 0.51 0.46 2.05 2.52 Methionine (5mM), Polysorbate 80 (1mg/mL), pH5.2 Formulation 18 Sucrose (55mg/mL), 0.15 0.38 0.53 0.45 2.04 2.49 Methionine (5mM), Polysorbate 20 (1mg/mL), pH5.2 Formulation 19 Sucrose (55mg/mL), 0.15 0.37 0.53 0.44 1.99 2.43 Methionine (5mM), Poloxamer 188 (1mg/mL), pH5.2 Referring to the s of Table 6, the contents of HMW and LMW before and after the storage at 40°C for 1 month did not vary icantly according to surfactant types.

That is, the stabilities of the formulation containing polysorbate 80, the formulation ning polysorbate 20, and the formulation containing poloxamer 188 were similar to one another.

Accordingly, it was confirmed that the s of polysorbate 80, polysorbate 20, and poloxamer 188 on the stability of adalimumab were r to one another. Formulations according to polyol types and comparison of stability of formulations f To e the stability of liquid formulations of adalimumab according to polyol types, formulations having the following compositions were prepared.

As in Formulation 14 of Example 2, Formulation 20 was prepared to contain sucrose (55 mg/mL), methionine (5 mM), polysorbate 80 (1 mg/mL), and adalimumab (100 mg/mL) (the same as Formulation 17 in Example 4). Additionally, Formulations 21 to 23 were ed by varying the polyol type to ose, PEG400, and PEG4000 while fixing the total amount of the polyol to be the same.

Each formulation was sterilized by filtration, filled into each 1 mL glass syringe in an amount of 0.4 mL, and stored at 40°C for 1 month. The amounts of impurities of HMW and LMW in samples before and after the storage were analyzed by SE-HPLC.

The compositions of Formulations 20 to 23 and the C contents of these formulations before and after the storage at 40°C for 1 month were analyzed and the results are shown in Table 7 below.

[Table 7] Before Storage at 40°C After Storage at 40°C for 1 Month for 1 Month HMW LMW Total HMW LMW Total (%) (%) (%) (%) (%) (%) Formulation Sucrose (55 mg/mL), Methionine (5 mM), and 0.14 0.37 0.51 0.46 2.05 2.52 Polysorbate 80 (1 mg/mL), pH 5.2 Formulation Trehalose (55 mg/mL), 0.14 0.36 0.50 0.46 2.03 2.49 21 Methionine (5 mM), and Polysorbate 80 (1 mg/mL), pH 5.2 ation PEG400 (55 mg/mL), 22 Methionine (5 mM), and 0.16 0.38 0.54 0.62 2.02 2.63 Polysorbate 80 (1 mg/mL), pH 5.2 ation PEG4000 (55 mg/mL), 23 Methionine (5 mM), and 0.17 0.37 0.53 0.85 2.05 2.90 Polysorbate 80 (1 mg/mL), pH 5.2 ing to the results of Table 7 above, it can be seen that the contents of HMW and LMW vary significantly ing to polyol types before and after the storage at 40°C for 1 month. The impurity contents of HMW and LMW of all of the formulations were r to one another before the storage at 40°C for 1 month. However, after the e at 40°C for 1 month, ations 20 to 22, which contained sucrose, trehalose, and PEG400, respectively, showed a relatively low HMW content compared to that of Formulation 23, which contained PEG4000. The LMW content was shown to be similar among these formulations. Accordingly, it was confirmed that there is a difference in stability among formulations depending on the polyol types used, and among these, sucrose and trehalose were shown to be more effective in improving stability than other types of polyols. Comparison of stability between ations consisting of polyol, arginine, methionine, surfactant, and additional stabilizer and HUMIRA® ation Samples were prepared through various compositions by varying the types of surfactants and polyols, presence/absence of an additional stabilizer, contents of arginine hydrochloride, etc. in the composition where 5 mM methionine was used as a stabilizer, and a composition in a commercial Humira® formulation was prepared. These samples were stored at 40°C and then subjected to SE-HPLC analysis to compare the stability among these formulations. The compositions for each of these formulations are shown in Table 8, and the contents of impurities before and after the storage at 40°C for 2 months are shown in Table 9.

[Table 8] Formulation No. Composition of Formulations Adalimumab (100 mg/mL), Methionine (5 mM), Arginine Hydrochloride (0 mM), Polysorbate 80 (1 mg/mL), e (55 mg/mL) Adalimumab (100 , Methionine (5 mM), Arginine Hydrochloride (20 mM), rbate 80 (1 mg/mL), Sucrose (55 mg/mL) Adalimumab (100 , Methionine (5 mM), Arginine 26 hloride (40 mM), Polysorbate 80 (1 mg/mL), e (55 mg/mL) Adalimumab (100 mg/mL), Methionine (5 mM), Arginine 27 Hydrochloride (40 mM), Poloxamer 188 (1 mg/mL), Sucrose (55 mg/mL), Adalimumab (100 mg/mL), Methionine (5 mM), Arginine 28 Hydrochloride (40 mM), Poloxamer 188 (1 mg/mL), Sucrose (55 mg/mL), Glycine (20 mM) Adalimumab (100 mg/mL), Methionine (5 mM), Arginine 29 Hydrochloride (40 mM), Poloxamer 188 (1 mg/mL), Sucrose (55 mg/mL), Leucine (20 mM) Adalimumab (100 mg/mL), Methionine (5 mM), Arginine Hydrochloride (40 mM), Poloxamer 188 (1 mg/mL), Sucrose (55 mg/mL), Glycine (10 mM), Leucine (10 mM) Adalimumab (100 mg/mL), nine (5 mM), Arginine 31 Hydrochloride (40 mM), Poloxamer 188 (1 mg/mL), Trehalose (55 mg/mL) Adalimumab (100 mg/mL), Methionine (5 mM), Arginine 32 Hydrochloride (40 mM), Poloxamer 188 (1 mg/mL), Trehalose (55 mg/mL), Glycine (20 mM) Adalimumab (100 mg/mL), Methionine (5 mM), Arginine 33 Hydrochloride (40 mM), Poloxamer 188 (1 mg/mL), Trehalose (55 mg/mL), Leucine (20 mM) Adalimumab (100 mg/mL), Methionine (5 mM), ne 34 Hydrochloride (40 mM), Poloxamer 188 (1 mg/mL), Trehalose (55 mg/mL), Glycine (10 mM), Leucine (10 mM) Adalimumab (100 mg/mL), Methionine (5 mM), ne Hydrochloride (40 mM), Poloxamer 188 (1 mg/mL), Sucrose (27.5 mg/mL), PEG4000 (27.5 mg/mL) umab (100 mg/mL), Methionine (5 mM), Arginine 36 Hydrochloride (40 mM), Poloxamer 188 (1 mg/mL), e (27.5 , PEG4000 (27.5 mg/mL), Glycine (20 mM) Adalimumab (100 mg/mL), Methionine (5 mM), Arginine 37 hloride (40 mM), Poloxamer 188 (1 mg/mL), e (27.5 mg/mL), PEG4000 (27.5 mg/mL), Leucine (20 mM) Adalimumab (100 mg/mL), Methionine (5 mM), Arginine Hydrochloride (40 mM), Poloxamer 188 (1 mg/mL), Sucrose (27.5 mg/mL), PEG4000 (27.5 mg/mL), Glycine (10 mM), Leucine (10 mM) Adalimumab (100 mg/mL), nine (5 mM), Arginine 39 Hydrochloride (40 mM), Poloxamer 188 (1 mg/mL), Trehalose (27.5 , PEG4000 (27.5 mg/mL) Adalimumab (100 mg/mL), Methionine (5 mM), Arginine 40 Hydrochloride (40 mM), Poloxamer 188 (1 mg/mL), Trehalose (27.5 mg/mL), PEG4000 (27.5 mg/mL), Glycine (20 mM) Adalimumab (100 mg/mL), Methionine (5 mM), Arginine 41 Hydrochloride (40 mM), Poloxamer 188 (1 mg/mL), Trehalose (27.5 mg/mL), PEG4000 (27.5 mg/mL), e (20 mM) Adalimumab (100 mg/mL), Methionine (5 mM), Arginine 42 Hydrochloride (40 mM), Poloxamer 188 (1 mg/mL), ose (27.5 mg/mL), PEG4000 (27.5 mg/mL), Glycine (10 mM), Leucine (10 mM) 43 Adalimumab (100 mg/mL), Commercial HUMIRA® Formulation Adalimumab (50 mg/mL), Methionine (5 mM), Arginine Hydrochloride (40 mM), Polysorbate 80 (1 mg/mL), Sucrose (55 mg/mL) 45 Adalimumab (50 mg/mL), Commercial HUMIRA® Formulation* * Commercial HUMIRA® Formulation: Sodium Phosphate Monobasic Dihydrate (0.86 mg/mL), Sodium Phosphate c Dihydrate (1.53 mg/mL), Sodium Citrate (0.3 mg/mL), Citric Acid Monohydrate (1.3 mg/mL), Mannitol (12 , Sodium Chloride (6.16 mg/mL), Polysorbate 80 (PS80) (1 mg/mL), pH 5.2 [Table 9] Formulation No. Before e at 40°C for 2 Months After Storage at 40°C for 2 Months HMW (%) LMW (%) Total (%) HMW (%) LMW (%) Total (%) 24 0.17 0.44 0.60 1.00 4.42 5.43 0.17 0.45 0.62 0.82 4.49 5.32 26 0.16 0.41 0.57 0.82 4.69 5.52 27 0.17 0.43 0.60 0.85 4.57 5.42 28 0.17 0.45 0.61 0.77 4.55 5.32 29 0.16 0.44 0.60 0.80 4.55 5.34 0.17 0.44 0.60 0.79 4.54 5.33 31 0.17 0.45 0.62 0.76 4.58 5.34 32 0.17 0.45 0.62 0.73 4.55 5.28 33 0.17 0.43 0.60 0.76 4.55 5.31 34 0.16 0.44 0.60 0.76 4.57 5.33 0.18 0.45 0.62 0.99 4.56 5.55 36 0.17 0.44 0.61 1.06 4.50 5.56 37 0.17 0.43 0.61 1.02 4.50 5.52 38 0.18 0.44 0.62 0.95 4.47 5.42 39 0.17 0.43 0.61 0.96 4.52 5.48 40 0.17 0.44 0.62 0.97 4.51 5.48 41 0.17 0.42 0.59 0.99 4.50 5.49 42 0.18 0.43 0.61 0.99 4.55 5.55 43 0.29 0.44 0.73 1.44 5.52 6.95 44 0.19 0.43 0.62 0.50 4.62 5.12 45 0.29 0.45 0.74 1.15 5.66 6.81 Referring to Table 8, sucrose, ose, a combination of sucrose and PEG4000, and a combination of trehalose and PEG4000 were used as polyols, whereas polysorbate 80 and poloxamer 188 were used as surfactants.

Arginine was used at a tration of 0 mM, 20 mM, or 40 mM, and to m the roles of additional additives, formulations were designed using glycine (Gly), leucine (Leu), or a ation of glycine and leucine, and the stability of the ing formulations of adalimumab (100 mg/mL) was compared to that of the commercial HUMIRA® formulation. Additionally, with respect to the stability of adalimumab (50 mg/mL), the commercial HUMIRA® formulation was compared to the composition of ation 44, which is the same as Formulation 26 except for the content of adalimumab.

As a result of the comparison of the stability among the formulations in Table 9, it was confirmed that all of the formulations had excellent stability compared to the commercial ® formulation. The total impurity ts before storage were in the range of 0.57 to 0.74 and were similar to one another. However, with respect to the stability of these formulations after the storage at 40°C for 2 months, it was med that other formulations had a lower impurity content of adalimumab compared to a commercial HUMIRA® ation having the adalimumab content of 100 mg/mL (Formulation 43) and a commercial HUMIRA® formulation having the adalimumab content of 50 mg/mL lation 45).

Accordingly, it was confirmed that the formulations of the present invention, which consist of various polyols and surfactants, arginine hydrochloride, and an additional stabilizer, are more stable than the commercial HUMIRA® formulations, with regard to the increase of impurities.

Additionally, while the total ty content was about 0.4% higher in the formulations of Example 5 after the storage at 40°C for 1 month when PEG 4000 was used as a polyol in Example 5 compared to the formulations where sucrose or trehalose was used as a polyol, in the formulations of this Example, the total impurity content of adalimumab when e or trehalose was used together with PEG4000, although the e period was 2 months, was shown to be similar to those of the formulations where sucrose or trehalose was used alone. Accordingly, it was confirmed that when an additional additive is added to obtain an additional effect such as antioxidation, etc., the replacement of part of sucrose or trehalose with PEG, which has a higher molecular weight, can be used as a method to maintain osmotic pressure and stability. Comparative ity evaluation of commercial ® formulation and formulations containing arginine hydrochloride in response to mechanical stress To compare the stability of the commercial HUMIRA® formulation and formulations containing arginine hydrochloride in response to mechanical stress, formulations with the following compositions were prepared and allowed to pass through a rotary piston pump, and thereby the numbers of particles before and after the pump passage were evaluated.

Additionally, to confirm whether the measured particles were derived from adalimumab, placebos were prepared by excluding only umab in the ition of each sample, allowed to pass through a rotary piston pump in the same conditions, and thereby the number of particles before and after the pump passage was evaluated. For the evaluation of the number of particles, a micro flow imaging device from Protein Simple was used. The composition for each sample is shown in Table 10, and the number of particles of each sample and a placebo thereof before and after the pump passage are shown in Table 11 and [Table 10] Formulation Adalimumab (100 mg/mL), Sucrose (55 mg/mL), ne Hydrochloride 46 (40 mM), Methionine (5 mM), Polysorbate 80 (1 mg/mL), pH 5.2 ation Adalimumab (50 mg/mL), e (55 mg/mL), Arginine Hydrochloride 47 (40 mM), Methionine (5 mM), rbate 80 (1 , pH 5.2 Adalimumab (50 mg/mL), Sodium Phosphate Monobasic Dihydrate Formulation (0.86 mg/mL), Sodium Phosphate Dibasic Dihydrate (1.53 mg/mL), Sodium Citrate (0.3 mg/mL), Citric Acid Monohydrate (1.3 mg/mL), Mannitol (12 mg/mL), Sodium Chloride (6.16 mg/mL), Polysorbate 80 (PS80) (1 mg/mL), pH 5.2 sition of Commercial HUMIRA®) [Table 11] le Concentration (#/mL) Placebo Sample (containing umab) Before Passing After Passing Before Passing After Passing Through Pump Through Pump Through Pump Through Pump Formulation 2667 3618 699 85743 Formulation 2667 3618 769 53734 Formulation 1213 7938 1917 150617 As shown in Table 10, Formulation 46 and ation 47 have the same additive composition, and the adalimumab content of Formulation 46 is 100 mg/mL and that of Formulation 47 is 50 mg/mL. In the case of Formulation 48, the composition and adalimumab content were adjusted to be the same as that of the commercial HUMIRA®.

Each formulation was passed through a rotary piston pump and the number of particles before and after the passage was analyzed. As a result, as shown in Table 11 and it was confirmed that Formulation 46 had 85,743 particles/mL and Formulation 47 had 53,734 particles/mL, and Formulation 46 with a high adalimumab content had a higher number of particles compared to that of ation 47 with a low adalimumab content.

In contrast, in the case of Formulation 48, i.e., the cial HUMIRA® formulation, the number of particles which passed through the pump was 150,617 particles/mL, thus showing a higher number of particles compared to those of Formulations 46 and 47. onally, the particle concentrations of the os of all of the formulations after pump passage were 3,618 particles/mL and 7,938 particles/mL, and thus it was confirmed that the particles measured in the samples containing adalimumab after the pump passage were derived from adalimumab. Accordingly, it was confirmed that the formulations containing arginine hydrochloride can effectively protect adalimumab in response to mechanical , compared to the commercial HUMIRA® formulation. Experiment of viscosity-lowering effect according to arginine hydrochloride concentration To examine the range of an arginine tration at which the ity an adalimumab solution can be lowered when arginine is added thereto, experiments were med as follows. As the control group containing no arginine hydrochloride (ArgHCl), samples were prepared to n adalimumab (100 mg/mL) and polysorbate 80 (1 mg/mL), or adalimumab (50 mg/mL) and polysorbate 80 (1 mg/mL). As the experimental group, samples containing arginine hydrochloride were ed by gradually increasing the concentration of arginine hydrochloride being added to each composition of the control group by 20 mM up to 180 mM (final concentration). The pH of all of the samples was about 5.2.

The viscosities of the prepared samples were measured using the mVROC instrument from Rheosense. The results of ity measurement are shown in Table 12.

[Table 12] ArgHCl Adalimumab (100 mg/mL) Adalimumab (50 mg/mL) (mM) (Unit: cp) (Unit: cp) 0 2.71 1.47 2.59 1.42 40 2.59 1.42 60 2.62 1.42 80 2.62 1.45 100 2.63 1.48 120 2.67 1.49 140 2.70 1.50 160 2.75 1.52 180 2.79 1.54 In Table 12 above, reviewing the viscosities of the formulations where adalimumab (100 mg/mL), polysorbate80 (1 mg/mL), and arginine hydrochloride (ArgHCl) at various concentrations were added, the viscosity of the solution containing adalimumab (100 mg/mL) and polysorbate 80 (1 mg/mL) without ArgHCl was 2.71 cp. When ArgHCl was added to the solution at a final concentration of 20 mM to 120 mM, it was confirmed that the ities of these compositions sed compared to that of the composition where ArgHCl was not added. In the case where ArgHCl was added at a final concenration of 140 mM, the ity of the ition was similar to that where ArgHCl was not added.

In the cases where ArgHCl was added at a final concentration of 160 mM or higher, the viscosity-lowering effect was not observed and the viscosities were increased compared to the ation where ArgHCl was not added.

In the case of the formulations containing adalimumab (50 mg/mL) and rbate 80 (1 mg/mL), the viscosity of the solution containing no ArgHCl was 1.47 cp, and in the cases where ArgHCl was added at a concentration of 20 mM to 80 mM, the viscosities were in the range of 1.42 to 1.45, thus being lower compared to the formulation where ArgHCl was not added. In the cases where ArgHCl was added at a final concentration of 100 mM, the viscosity was similar to that where ArgHCl was not added, and in the cases where ArgHCl was added at a final concentration of 120 mM or higher, the ities were increased compared to the case where ArgHCl was not added.

Accordingly, it was confirmed that in the case of a solution of adalimumab (100 mg/mL), the viscosity can be lowered by adding thereto ArgHCl at a concentration of 140 mM or less, and in the case of a solution of adalimumab (50 mg/mL), the viscosity can be lowered by adding thereto ArgHCl at a concentration of 100 mM or less. Experiment 1 confirming effects of buffers and salts in adalimumab formulations To determine the effects of buffers and salts on the stability of adalimumab, samples of formulations without buffers and salts were prepared, and samples of formulations where a buffer or salt was added to the above formulations were prepared, and these formulations were stored at 40°C for 2 weeks and 1 month. The ity of these samples was ed by SE-HPLC and the pH of each sample was measured.

[Table 13] For After Storage at After Storage at 40°C Additional Zerotime mula 40°C for 2 Weeks for 1 Month Formulation Buffer and Salt HMW LMW Total HMW LMW Total HMW LMW Total A-1 Adalimumab No Buffer/No 0.36 0.38 0.74 0.47 1.19 1.66 0.65 2.04 2.69 (100 mg/mL), Salt A-2 PS80 Sodium (1 mg/mL), ate/ pH 5.2 Sodium Citrate 0.38 0.39 0.76 0.59 1.25 1.84 0.92 2.27 3.19 (Humira Composition Buffer)* A-3 NaCl 0.37 0.41 0.78 0.66 1.42 2.09 0.86 2.50 3.35 (100 mM) A-4 Ammonium e 0.37 0.41 0.78 0.50 1.42 1.92 0.69 2.54 3.24 (100 mM) A-5 Sodium Sulfate 0.39 0.41 0.80 0.54 1.41 1.95 0.75 2.49 3.24 (100 mM) A-6 Adalimumab No Buffer/No 0.37 0.40 0.77 0.28 1.21 1.49 0.38 2.03 2.41 (50 mg/mL), Salt A-7 PS80 Sodium (1 mg/mL), Phosphate/ pH 5.2 Sodium Citrate 0.38 0.41 0.79 0.43 1.27 1.69 0.66 2.20 2.86 (Humira Composition Buffer)* A-8 NaCl 0.38 0.42 0.80 0.43 1.38 1.81 0.63 2.50 3.13 (100 mM) A-9 Ammonium Sulfate 0.39 0.41 0.80 0.38 1.45 1.83 0.53 2.58 3.10 (100 mM) A-10 Sodium e 0.39 0.44 0.83 0.40 1.45 1.85 0.55 2.51 3.05 (100 mM) *Humira® Composition Buffer: sodium phosphate monobasic dihydrate (0.86 mg/mL), sodium phosphate dibasic dihydrate (1.53 mg/mL), sodium citrate (0.3 mg/mL), citric acid monohydrate (1.3 mg/mL) Table 13 shows the results of SE-HPLC analysis of the ations of the samples at zero-time of each sample, after the storage at 40°C for 2 weeks and after the e at 40°C for 1 month. Formulation A-1 was prepared to contain adalimumab (100 mg/mL) and PS80 (1 mg/mL), and Formulations A-2 to A-5 were prepared to contain a buffer or salt in Formulation A-1. Additionally, Formulations A-6 to A-10 were prepared by adjusting the adalimumab concentration of A-1~A-5 composition to 50 mg/mL. Reviewing the results of SE-HPLC analysis of each sample, when Formulation A-1 without a buffer and Formulation A-2 with a buffer were compared (or Formulations A-6 and A-7 with different concentrations of adalimumab were compared), it was found that the formulations without a buffer had a smaller increase in HMW and LMW, thus ming that these formulations are more stable.

Additionally, when ations A-1 and A-6 and formulations containing a salt including NaCl, um sulfate, sodium sulfate, etc. were compared, it was found that formulations without a salt had a smaller increase in HMW and LMW compared to ations with a salt. ingly, it was confirmed that umab formulations are preferred not to contain a salt from the aspect of stability.

Additionally, all of the samples were maintained to have a constant pH of 5.2 before storage, 2 weeks after storage, and 1 month after storage. Accordingly, it was confirmed that adalimumab at a concentration of 50 mg/mL or higher has a sufficient intrinsic buffering effect, thus not requiring the use of a buffer, and by not using a buffer, formulations with improved stability can be constituted. Experiment 2 confirming effects of s and salts in adalimumab formulations To ine the effects of buffers and salts on the stability of adalimumab, samples of formulations ting of adalimumab (100 mg/mL or 50 mg/mL), stabilizer se (55 mg/mL) or glycine (160 mM)), arginine hydrochloride l: 50 mM), methionine (5 mM), and polysorbate 80 (1 mg/mL) were prepared, and samples of formulations where a buffer or salt was added to the above formulations were prepared. For comparison purposes, samples with the Humira® composition containing adalimumab (100 mg/mL) or adalimumab (50 mg/mL) were prepared, and each formulation was filled into a glass syringe in an amount of 0.4 mL per syringe, stored at 55°C for one week, and the stability was compared by SE-HPLC and the pH was measured. The s of each composition and the monomer contents before and after storage at 55°C for 1 week are shown in Table 14 below.

[Table 14] r Content (%) Before After Formulation Additional Buffer and Salt Storage at Storage at 55°C for 1 55°C for 1 Week Week A-11 Adalimumab - 98.9 94.8 A-12 (100 mg/mL), Sodium Chloride (NaCl) (100 mM) 98.8 94.7 A-13 Sucrose Ammonium Sulfate (100 mM) 98.9 94.6 A-14 (55 mg/mL), Magnesium Chloride (MgCl2) (100 mM) 98.9 94.7 A-15 ArgHCl Calcium Chloride (CaCl2) (100 mM) 98.8 94.7 (50 mM), Met (5 mM), PS80 A-16 Sodium Acetate (20 mM) 98.9 94.1 (1 mg/mL), pH 5.2 A-17 Adalimumab - 98.9 95.2 A-18 (100 mg/mL), Sodium Chloride (100 mM) 98.9 94.8 A-19 Gly (160 mM), Ammonium Sulfate (100 mM) 98.9 94.8 A-20 ArgHCl Magnesium Chloride (100 mM) 98.9 94.8 A-21 (50 mM), Met Calcium Chloride (100 mM) 98.8 94.6 (5 mM), PS80 Sodium Phosphate/Sodium Citrate A-22 98.9 94.9 (1 mg/mL), (Humira® Composition Buffer)** A-23 pH 5.2 Sodium Acetate (20 mM) 98.9 94.0 A-24 umab (100 mg/mL), Humira® Composition* 98.8 93.9 A-25 Adalimumab - 99.0 95.2 A-26 (50 , Sodium Chloride (100 mM) 98.9 94.8 A-27 Sucrose Ammonium Sulfate (100 mM) 98.9 94.8 A-28 (55 mg/mL), Magnesium Chloride (100 mM) 98.9 95.0 A-29 ArgHCl Calcium Chloride (100 mM) 98.9 94.4 (50 mM), Met Sodium Phosphate/Sodium Citrate A-30 98.9 94.9 (5 mM), PS80 (Humira® Composition Buffer)** (1 mg/mL), A-31 Sodium e (20 mM) 98.9 93.5 pH 5.2 A-32 umab - 99.0 95.6 A-33 (50 mg/mL), Sodium Chloride (100 mM) 98.9 95.4 A-34 Gly (160 mM), Ammonium Sulfate (100 mM) 98.9 95.0 A-35 ArgHCl ium Chloride (100 mM) 98.9 94.9 A-36 (50 mM), Met Calcium Chloride (100 mM) 98.8 94.7 (5 mM), PS80 Sodium Phosphate/Sodium Citrate A-37 98.9 95.3 (1 mg/mL), (Humira® Composition )** A-38 pH 5.2 Sodium Acetate (20 mM) 98.9 93.6 A-39 Adalimumab (50 mg/mL), Humira® Composition* 98.9 93.6 * Humira® Composition: Sodium phosphate monobasic dihydrate (0.86 mg/mL), Sodium ate dibasic dihydrate (1.53 mg/mL), Sodium citrate (0.3 mg/mL), Citric acid monohydrate (1.3 mg/mL), Mannitol (12 mg/mL), Sodium chloride (6.16 mg/mL), PS80 (1 mg/mL) ** ® Composition Buffer: Sodium phosphate monobasic dihydrate (0.86 mg/mL), Sodium phosphate dibasic dihydrate (1.53 mg/mL), Sodium citrate (0.3 mg/mL), Citric acid monohydrate (1.3 mg/mL) First, all of the samples were maintained to have a constant pH of 5.2 before and after storage at 55°C for 1 week. Accordingly, it was confirmed that adalimumab at a concentration of 50 mg/mL or higher has a sufficient intrinsic buffering effect in the itions and similar compositions thereof in Table 14 above.

As shown in Table 14 above, the monomer contents of the samples were similar to one another, in the range of 98.8% to 99.0%. After the storage at 55°C for 1 week, the monomer contents were shown to be different according to compositions. The monomer content of sample A-11, with a composition of adalimumab (100 mg/mL), e (55 mg/mL), ArgHCl (50 mM), methionine (5 mM), and polysorbate 80 (1 mg/mL), was 94.8% after the storage. However, in the cases of samples A-12 to A-16, where the salts of sodium chloride, ammonium sulfate, magnesium de, and calcium chloride, or a sodium citrate buffer were used, the monomer contents after the storage were in the range of 94.1% to 94.7%, which was lower ed to that of the sample with a composition where a salt and a buffer were not used. In the case of sample A-17, where Gly (160 mM) was used as a stabilizer instead of sucrose, the monomer content after the storage was 95.2%; however, in the cases of samples A-18 to A-23, where salts and buffers were used, the monomer contents after the storage were in the range of 94.0% to 94.9%, thus being lower compared to that of the sample with a composition where a salt and a buffer were not used. Even in the case where the ition was prepared by lowering the content of adalimumab to 50 mg/mL, when sucrose was used as a stabilizer without using a salt and a buffer (i.e., sample A-25), the monomer content after the storage was 95.2%, and when e (Gly) was used as a stabilizer without using a salt and a buffer (i.e., sample A-32), the monomer content after the storage was 95.6%. However, when additional salt/buffer was used in each formulation, in the cases of formulations (samples A-26 to A-31) where sucrose was contained, the r ts after the storage were in the range of 93.5% to 95.0%, and in the cases of formulations (samples A-33 to A-38) where Gly was contained, the monomer contents after the storage were in the range of 93.6% to 95.4%, thus showing that the monomer contents of the ations where additional salt/buffer was used was lower compared to that of the formulation where a salt and a buffer were not used. Accordingly, it was confirmed that when additional salt and buffer are not used in the formulations where adalimumab, arginine, stabilizer, and surfactants are contained, the stability of adalimumab can be improved, and that it is le to prepare a composition with ed ity using the intrinsic buffering effect of adalimumab itself, t using an additional buffer. r, when compared to samples A-24 and A-39, with the Humira® composition having the same adalimumab content, the monomer contents of the formulations where a salt and a buffer were contained, after the storage at 55°C for 1 week, were greater compared to those of the samples with the Humira® composition after the storage at 55°C for 1 week.

Accordingly, it is preferable from the aspect of stability that additional salt and buffer not be used in adalimumab formulations where arginine, tants, and stabilizers are contained; however, it was confirmed that the formulations containing arginine, surfactants, and stabilizers are more stable compared to the commercial Humira® formulations, regardless of whether these formulations contain additional salt and buffer. ment for confirming stabilizing effect by polyols in adalimumab formulations To compare the izing effect of polyols, which are used to improve the stability of adalimumab in a on, an adalimumab solution (112 mg/mL adalimumab) and a formulation ning adalimumab at a concentration of 112 mg/mL and each kind of polyol at a concentration of 42 mg/mL were prepared as follows. The contents of HMW, LMW, and monomers were ed using the SE-HPLC after repeating the freezing/thawing process 5 cycles and 10 cycles at -70°C and 5°C, respectively.

[Table 15] Formulation Zerotime FT 5C FT 10C HMW LMW Total HMW LMW Total HMW LMW Total (%) (%) (%) (%) (%) (%) (%) (%) (%) No Polyol 0.40 0.42 0.82 1.63 0.49 2.13 2.08 0.37 2.45 Mannitol (42 mg/mL) 0.39 0.41 0.80 0.51 0.48 0.99 0.65 0.36 1.01 Sucrose (42 mg/mL) 0.38 0.41 0.79 0.35 0.47 0.82 0.34 0.34 0.68 Trehalose (42 mg/mL) 0.38 0.41 0.79 0.35 0.47 0.82 0.35 0.35 0.70 Table 15 shows formulations of samples, and the results of SE-HPLC analysis of samples ing to sampling points of stability test. In the above results, when each sample was ted to a process of repeated freezing/thawing, the increase of HMW and LMW in the formulations where mannitol, sucrose, or trehalose was added tended to se compared to the formulation where no polyol was added, thus confirming the presence of a stabilizing effect by polyols. Comparing the stabilizing effect by each polyol ing to its type, when sucrose or trehalose was added, the contents of HMW and LMW were similar to those of the samples before undergoing a freezing/thawing process, and additionally, the purities were shown to be similar to those before a ng/thawing process even after they underwent 10 cycles of the freezing/thawing process, thus ming the significant izing effect by polyols. In contrast, in the case of the sample containing mannitol, when the sample was subjected to a process of repeated freezing/thawing, it was confirmed that the HMW content tended to increase and the purity of the sample was lowered during the freezing/thawing s. Accordingly, it was confirmed that sucrose and trehalose have a greater stabilizing effect compared to mannitol. Freezing/thawing experiment of adalimumab formulations for confirmation and comparison of stabilizing effects by ne, methionine, glycine, and sucrose To confirm the effects of arginine, methionine, glycine, and sucrose on stabilization during a freezing/thawing process of stock solutions, s were prepared as shown below.

Each sample in an amount of 1 mL was added into a polycarbonate bottle (5 mL), subjected to 5 cycles of the freezing/thawing process upon ation, and at -70°C and 5°C, respectively, and analyzed using SE-HPLC. The composition of each sample and SE-HPLC results of each sample before and after the freezing/thawing process are shown below.

[Table 16] Additional Stabilizer Before After 5 Cycles of Composition ArgHCl Gly Met sucrose Freezing/Thawing Freezing/Thawing (mM) (mM) (mM) (mg/mL) HMW LMW Total HMW LMW Total Adalimumab - - - - 0.32 0.40 0.72 1.76 0.28 2.04 (130 mg/mL), 50 - - - 0.36 0.39 0.74 0.67 0.31 0.98 Polysorbate 80 50 0.34 0.37 0.71 0.67 0.33 1.00 (1 mg/mL) 50 0.33 0.34 0.67 0.52 0.33 0.85 50 160 5 - 0.34 0.36 0.70 0.41 0.31 0.73 50 140 25 - 0.33 0.34 0.66 0.43 0.33 0.75 50 0 5 55 0.34 0.35 0.69 0.41 0.29 0.70 Samples were prepared to contain adalimumab (130 mg/mL) and polysorbate 80 (1 mg/mL), and to contain an additional stabilizer thereto. The contents of impurities of each sample before a freezing/thawing process were shown to be similar to one r both in HMW and LMW. After the freezing/thawing process, the LMW content was shown to be similar in all of the samples; however, the HMW content was shown to vary according to the type and content of the additional stabilizer. In the case of a sample where an additional izer was not added, the HMW content after undergoing 5 cycles of the freezing/thawing process sed up to 1.76%, s in the case of a sample containing ne hydrochloride (50 mM), the HMW content after undergoing 5 cycles of the freezing/thawing process icantly decreased to 0.67%. In the case of a formulation where methionine was added in addition to arginine hydrochloride (50 mM), an additional decrease of the HMW content was not confirmed when the formulation contained methionine at a concentration of 5 mM, whereas the HMW content was onally decreased by about 0.52% when the formulation contained methionine at a concentration of 25 mM. In the cases where glycine or sucrose was added in addition to ne hydrochloride and methionine, it was confirmed that the HMW content after undergoing 5 cycles of the ng/thawing process decreased further to be in the range of 0.41% to 0.43%, and both the formulation containing glycine and the formulation containing sucrose exhibited similar stability.

Accordingly, it was confirmed that all of methionine, arginine, glycine, and sucrose contribute to the stability of adalimumab. Additionally, it was confirmed that similar levels of stability could be attained when formulations are prepared by appropriate combinations using a polyol or amino acid as a stabilizer. Comparative ments on stability among ations containing glycine, formulations containing sucrose, and commercial Humira® formulations To compare the stability of formulations containing glycine, formulations containing sucrose, and commercial Humira® formulations, samples were prepared to contain umab at a concentration of 100 mg/mL or 50 mg/mL, arginine hydrochloride (ArgHCl) at a tration of 50 mM, polysorbate 80 (PS80) at a concentration of 1 mg/mL, and methionine at a concentration of 5 mM, and then, additional samples were prepared by adding glycine, a combination of glycine and methionine, or sucrose as an additional stabilizer to the above compositions. Additionally, samples were prepared to have commercial Humira® compositions and contain adalimumab at a tration of 100 mg/mL or 50 mg/mL. Each of the above samples was filled into 1 mL glass syringes in an amount of 0.4 mL per syringe. After storing each syringe at 40°C for 4 weeks, the HMW and LMW contents of each sample were analyzed by SE-HPLC. The composition of each composition and the results of C are shown in Table 17 below.

[Table 17] Before Storage at 40°C After e at 40°C Composition Additional Stabilizer for 4 Weeks for 4 Weeks HMW LMW Total HMW LMW Total - 0.28 0.39 0.67 0.54 2.44 2.99 Gly (120 mM) 0.27 0.41 0.68 0.56 2.42 2.99 Gly (160 mM) 0.28 0.42 0.69 0.54 2.37 2.91 Adalimumab Gly (100 mM), Met (100 mg/mL), 0.26 0.40 0.67 0.54 2.40 2.94 (20 mM) Arg·HCl (50 mM), Gly (120 mM), Met PS80 (1 mg/mL), 0.27 0.39 0.65 0.53 2.46 2.99 (20 mM) Met (5 mM) Gly (140 mM), Met 0.28 0.43 0.70 0.49 2.32 2.81 (20 mM) Sucrose (55 mg/mL) 0.27 0.40 0.67 0.48 2.51 2.99 Adalimumab (100 mg/mL), Humira® 0.40 0.40 0.80 0.98 3.05 4.03 Composition Adalimumab - 0.27 0.38 0.64 0.35 2.36 2.71 (50 mg/mL), Gly (120 mM) 0.26 0.38 0.64 0.35 2.29 2.64 Arg·HCl (50 mM), Gly (160 mM) 0.28 0.43 0.71 0.32 2.26 2.58 PS80 (1 mg/mL), Gly (100 mM), Met 0.26 0.39 0.64 0.33 2.33 2.66 Met (5 mM) (20 mM) Gly (120 mM), Met 0.25 0.36 0.61 0.34 2.38 2.71 (20 mM) Gly (140 mM), Met 0.27 0.43 0.70 0.30 2.26 2.56 (20 mM) Sucrose (55 mg/mL) 0.26 0.38 0.64 0.31 2.33 2.64 Adalimumab (50 mg/mL), Humira® 0.43 0.41 0.84 0.82 3.23 4.06 composition The s with compositions ning arginine hydrochloride, polysorbate 80, methionine, and adalimumab showed similar levels of stability before the e at 40°C for 4 weeks. Meanwhile, the samples with the commercial Humira® itions showed a higher HMW content by about 0.1% upon preparation and comparison to other samples. With regard to the sum of the HMW and LMW contents after the storage, all of the samples containing arginine hydrochloride showed relatively low values: that is, 2.81% to 2.99% when adalimumab was contained at a concentration of 100 mg/mL; and 2.56% to 2.71% when umab was contained at a concentration of 50 mg/mL. However, in the case of commercial Humira® compositions, the sum of the HMW and LMW ts after the storage was 4.03% when adalimumab was contained at a concentration of 100 mg/mL; and 4.06% when adalimumab was contained at a concentration of 50 mg/mL, thus showing higher levels of HMW and LMW ts, ed to the samples with compositions containing arginine.

Accordingly, it was confirmed that the formulations containing combinations of additives bed in Examples of the present invention; that is, the formulations containing arginine and formulations containing a polyol or amino acid as an additional stabilizer are superior to the commercial Humira® formulations with respect to stability. Comparative experiments on stability of adalimumab formulations according to concentration of adalimumab, sucrose, glycine, leucine, methionine, sodium chloride (NaCl), and ne To compare the stability of adalimumab formulations, samples of various compositions were prepared by a combination of adalimumab, arginine hydrochloride (ArgHCl), sodium chloride (NaCl), polysorbate 80 (PS80), nine (Met), sucrose, glycine (Gly), and leucine (Leu). Additionally, samples were prepared to have commercial Humira® compositions and contain adalimumab at a concentration of 100 mg/mL or 50 mg/mL for comparison es. Each formulation was filled into a 1 mL glass syringe in an amount of 0.4 mL, stored at 40°C for 4 weeks, and the monomer contents before and after the storage were ed by SE-HPLC. The compositions and the SE-HPLC results are shown below.

[Table 18] Monomer Composition Content (%) Before After Storag Storag Adalimu NaC Sucros ArgH Met PS80 Gly Leu e at e at mab l e Cl (m (mg/m (m (m (mg/mL (m (mg/m 40°C 40°C (mM) M) L) M) M) ) M) L) for 4 for 4 Weeks Weeks 100 50 5 1 55 99.29 97.15 100 50 5 1 45 99.33 97.16 100 50 25 1 45 99.30 97.14 100 50 5 1 45 20 99.30 97.18 100 50 5 1 45 20 99.30 97.12 100 50 5 1 35 99.30 97.06 100 50 25 1 35 99.30 97.14 100 50 25 1 35 40 99.33 97.09 100 50 25 1 35 20 20 99.29 97.23 100 50 5 1 25 99.31 97.11 100 50 25 1 25 99.32 97.13 100 50 25 1 25 60 99.30 97.17 100 50 25 1 25 40 20 99.31 97.17 100 50 5 1 99.31 97.06 100 50 25 1 99.32 97.13 100 50 25 1 140 99.30 97.04 100 50 50 5 1 25 99.31 96.96 100 50 50 15 1 25 99.28 96.93 100 50 50 25 1 25 99.27 96.89 100 25 60 5 1 25 99.30 96.88 100 25 60 35 1 25 99.29 97.00 100 25 60 5 1 35 99.27 96.85 100 Commercial ® Formulations 99.29 95.64 50 50 5 1 55 99.30 97.23 50 50 5 1 45 99.30 97.21 50 50 25 1 45 99.31 97.28 50 50 5 1 45 20 99.30 97.30 50 50 5 1 45 20 99.30 97.32 50 50 5 1 35 99.30 97.23 50 50 25 1 35 99.31 97.23 50 50 25 1 35 40 99.30 97.27 50 50 25 1 35 20 20 99.30 97.34 50 50 5 1 25 99.31 97.15 50 50 25 1 25 99.30 97.24 50 50 25 1 25 60 99.30 97.32 A-7 50 50 25 1 25 40 20 99.31 97.26 50 50 5 1 99.29 97.25 50 50 25 1 99.30 97.33 50 50 25 1 140 99.29 97.33 50 50 25 1 120 20 99.31 97.38 50 50 50 5 1 25 99.29 97.06 50 50 50 15 1 25 99.30 96.90 50 50 50 25 1 25 99.27 97.02 50 25 60 5 1 25 99.30 96.81 50 25 60 35 1 25 99.30 97.00 50 25 60 5 1 35 99.29 96.92 50 Commercial Humira® Formulations 99.29 95.47 * Commercial Humira® Formulations: Sodium phosphate monobasic dihydrate (0.86 mg/mL), Sodium phosphate dibasic dihydrate (1.53 mg/mL), Sodium e (0.3 mg/mL), Citric acid monohydrate (1.3 mg/mL), Mannitol (12 mg/mL), Sodium chloride (6.16 mg/mL), PS80 (1 mg/mL) The monomer contents of all of the samples before the storage at 40°C were similar to one r, in the range of 99.27% to , regardless of the formulations. With regard to the monomer ts of samples (A-40 to A-61) containing adalimumab (100 mg/mL) after the storage at 40°C for 4 weeks, in the case of samples (A-40 to A-55) where sodium chloride was not contained, the r contents were in the range of 97.04% to 97.23%, whereas in the case of samples ning sodium chloride (A-56 to A-61 excluding A-62 which is a ® composition), the monomer contents were in the range of 96.85% to 97.00%, thus showing slightly lower monomer contents compared to those of the formulations where sodium chloride was not contained. However, the difference in monomer contents between these samples according to the contents of sodium chloride, e, methionine, glycine, and leucine after the storage at 40°C for 4 weeks was shown to be relatively insignificant, and the monomer contents of the compositions (A-40 to A-61) were shown to be higher than 95.64%, the monomer content of A-62, which is a Humira® ition containing adalimumab at the same concentration, by at least 1%.

In the case where adalimumab was contained at a concentration of 50 mg/mL, the results of analysis were the same as in the compositions where adalimumab was ned at a concentration of 100 mg/mL. With regard to the monomer ts of the samples containing adalimumab (50 mg/mL) (A-63 to A-85) after the storage at 40°C for 4 weeks, in the case of samples A-63 to A-79, where sodium chloride was not contained, the monomer contents were in the range of 97.15% to 97.38%, whereas in the case of samples containing sodium chloride (A-80 to A-85, excluding A-86, which is a ® composition), the monomer contents were in the range of 96.81% to 97.06%, thus showing slightly lower monomer contents compared to those of the formulations where sodium chloride was not contained. However, the difference in monomer contents between these samples ing to the contents of sodium chloride, sucrose, methionine, glycine, and leucine after the storage at 40°C for 4 weeks was shown to be relatively insignificant, and the monomer contents of the compositions (A-63 to A-85) were shown to be higher than 95.47%, the monomer content of A-86, which is a Humira® composition ning adalimumab at the same concentration, by at least 1%.

Accordingly, it was confirmed that the combinations of additives and formulations f described in es of the present invention, that is, the formulations containing arginine, are superior to the commercial ® formulations with respect to stability.

From the ing, a skilled person in the art to which the present invention pertains will be able to understand that the present invention may be embodied in other specific forms without modifying the technical concepts or essential characteristics of the present invention.

In this regard, the exemplary embodiments disclosed herein are only for illustrative purposes and should not be construed as limiting the scope of the present invention. On the contrary, the present invention is intended to cover not only the exemplary embodiments but also various alternatives, cations, equivalents, and other embodiments that may be included within the spirit and scope of the present invention as defined by the appended claims.

Claims (1)

1. A liquid ation of an adalimumab comprising adalimumab at a tration of 50 mg/mL to 130 mg/mL, a stabilizer, a surfactant at a concentration of 0.5 mg/mL to 1.5 mg/mL, a methionine at a concentration of 2.5 mM to 35 mM, and arginine at a concentration of 20 mM to