JP2020510079A - Liquid formulation of anti-TNFα antibody - Google Patents

Abstract

The present invention relates to anti-TNFα antibodies, in particular liquid formulations of adalimumab.

Description

  The present invention relates to a liquid formulation of an anti-TNFα antibody, specifically, adalimumab.

  Tumor necrosis factor α (TNF alpha, TNFα) is a cytokine produced by various cells such as mononuclear leukocytes and macrophages upon stimulation with endotoxin and the like. TNFα is a major mediator of major inflammatory, immunological, and pathophysiological responses that play a role in inducing responses such as T cell activation and thymocyte proliferation by activating TNF receptors (Non-Patent Document 1). ).

  Adalimumab is a recombinant human immunoglobulin G1 monoclonal antibody that selectively binds to tumor necrosis factor α in the body, thereby inhibiting the immune response by tumor necrosis factor α. Adalimumab was developed by BASP Bioresearch Corporation around 1993, and a therapeutic agent for rheumatoid arthritis manufactured by Abbott Laboratories was approved. Adalimumab is sold under the trade name Humira, which is licensed for the treatment of rheumatoid arthritis and is used in the treatment of Crohn's disease, ankylosing spondylitis, psoriatic arthritis, ulcerative colitis and the like.

  Adalimumab is the first fully human antibody developed as a pharmaceutical, developed by applying the phage display method, and has increased affinity by mutating CDRs. Adalimumab, also called D2E7, has a molecular weight of about 148 kD consisting of 1330 amino acids (Patent Document 1). Adalimumab is a TNFα inhibitor that inhibits the TNF-induced response by binding to TNF and preventing the TNF receptor and TNF on the p55 and p75 cell surfaces from reacting with each other.

  On the other hand, an antibody drug is a type of protein drug, and physical and chemical denaturation can occur due to various factors. Examples of protein denaturation include chemical denaturation such as oxidation, deamidation, and isomerization, and structural denaturation such as fragmentation and aggregation.When a protein is denatured, the protein loses its pharmacological activity, An unwanted immune response may be induced in the body as a side effect. When the antibody is fragmented, the binding affinity and the retention time in the body change, which may affect the pharmacological activity. In addition, there is a study that fragmented antibodies induce antibody aggregation. Furthermore, pharmacological activity may be reduced by aggregation. When comparing commercially available interferon β products, there is a research result that a product having a high content of aggregates and particles has a high ratio of producing neutralizing antibodies in the body (Non-Patent Document 2). . When a neutralizing antibody is produced in the body, even when a protein drug is injected, the neutralizing antibody binds to the protein drug, thereby affecting the safety, pharmacological effects, and pharmacokinetics of the protein drug. Also, denaturation and aggregation of Epoetin Alfa has been disclosed as a cause of the enhanced immunogenicity of the pharmaceutical Epoetin Alfa. Therefore, it is very important to produce a protein drug in an appropriate dosage form so as not to lose its physiological activity during the storage period and to prevent the protein from being fragmented, aggregated, or particulated. . Therefore, studies on dosage forms of various protein drugs have been actively conducted.

U.S. Pat. No. 6,090,382

Grell, M., et al. (1995) Cell, 83: 793-802 Barnard et al., 2013, J. Pharm. Sci. 102: 915

  The study of protein dosage forms aims to find the optimal combination by properly mixing various additives in consideration of the characteristics of each product, and to ensure stable storage until administered to patients. And The main purpose of adding the additives is to stabilize the protein and adjust the physical properties of the mixture. Additives are classified into surfactants, stabilizers, preservatives, buffers, isotonic agents, and the like, depending on their purpose and properties. In an antibody drug, it is necessary to administer a larger amount of protein than other protein drugs in order to obtain an effective therapeutic effect. In addition, when the administration route is subcutaneous injection, it is important to develop a high-concentration dosage form because an attempt to administer a large amount at one time may cause pain for the patient and inconvenience in production. As the concentration of the protein increases, intermolecular interaction increases, which causes problems such as increase in aggregates, increase in viscosity, gelation of the solution, and precipitation. When the viscosity is excessively increased, production is not easy, and administration to a patient becomes difficult due to increased injection pressure. Therefore, various methods for predicting and lowering the viscosity of a high-concentration antibody solution have been studied.

  An object of the present invention is to provide a liquid preparation of an anti-TNFα antibody.

  Another object of the present invention is to provide a method for producing the liquid preparation.

  Furthermore, an object of the present invention is to provide a method for increasing the stabilization of an anti-TNFα antibody using a composition comprising a stabilizer, a surfactant and arginine.

  Furthermore, another object of the present invention is to provide a method for increasing the stabilization of an anti-TNFα antibody using a composition that does not contain a buffer and contains a stabilizer, a surfactant, and arginine.

  The liquid formulation of the anti-TNFa antibody, in particular adalimumab, according to the present invention allows for long-term storage as it reduces by-product formation of adalimumab during storage. Further, since the denaturation and aggregation are prevented from occurring due to physical stress during the production process and transportation, the pharmacological efficacy of adalimumab can be stably stored for a long period of time. Therefore, it is expected that the liquid preparation according to the present invention can be effectively applied in the therapeutic field related to the pharmacological efficacy of adalimumab in the future.

FIG. 3 is a diagram showing the viscosity of each sample of Example 1 in the additive composition. It is a figure which shows the particle number of the sample of Example 7, and placebo before and after passing a pump.

  Hereinafter, the present invention will be described in more detail.

  Note that each description and embodiment disclosed in the present invention is also applied to each other description and embodiment. That is, any combination of various elements disclosed in the present invention is included in the present invention. Further, the present invention is not limited to the following specific description.

  Also, one of ordinary skill in the art can recognize and confirm the many equivalents of the specific embodiments of the invention described in the invention using only routine experimentation. There will be. Moreover, such equivalents are intended to be included in the present invention.

  One embodiment of the present invention for solving the above problem is a liquid preparation of an anti-TNFα antibody.

  The “anti-TNFα antibody” in the present invention means an antibody that binds to TNFα and regulates its biological activity. More specifically, the antibody has a function of suppressing signal transduction by TNFα by binding to TNFα and suppressing the binding between TNFα and its receptor. Further, such an anti-TNFα antibody may be a monoclonal antibody.

  The anti-TNFα antibody may be in the form of a full-length antibody or an antibody fragment containing an antigen-binding site thereof, but is not particularly limited thereto.

  More specifically, the anti-TNFα antibody may be a recombinant human immunoglobulin G1 monoclonal antibody, and more specifically, adalimumab. Information on adalimumab can be easily obtained by those skilled in the art from a known database.

  The antibody can be produced by a recombinant DNA technique using a mammalian cell expression system, but is not particularly limited thereto.

  The antibody may be included in the liquid formulation according to the present invention in a therapeutically effective amount. For example, a concentration of 1 to 250 mg / mL, specifically a concentration of 20 to 200 mg / mL, more specifically a concentration of 50 mg / mL to 200 mg / mL, more specifically 50 mg / mL, 100 mg / mL or It may be present at a concentration of 130 mg / mL, but is not limited thereto.

  The liquid preparation of the present invention may contain a stabilizer, a surfactant, and arginine in addition to the anti-TNFα antibody. The liquid formulation may be in the form of a solution that can stably store the anti-TNFα antibody.

  Specifically, in order to measure the stability of the anti-TNFα antibody, a protein stability analysis method known in the art may be used. Stability may be measured at a particular time at a particular temperature. For rapid testing, the dosage form can be stored at a higher or "accelerated" temperature, e.g., 40 <0> C for 2 weeks to 1 month or more, at which point time dependent stability is measured.

  In the present invention, “to impart stabilization to an anti-TNFα antibody” means that the loss of the active ingredient is less than a specific amount, for example, less than 10% under a specific storage condition, specifically, at a specific temperature for a predetermined time. means. Usually, 90% or more, specifically, about 92% or more of anti-TNFα antibody remains at 5 ± 3 ° C. for 2 years, at 25 ± 2 ° C. for 6 months, or at 40 ± 2 ° C. for 1 to 2 months. If the rate is maintained, the formulation is considered stable.

  The stabilizer contained in the liquid preparation of the present invention may be a polyol, an amino acid or a combination thereof. Here, the amino acid may be another amino acid other than arginine.

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

  More specifically, the polyol may be mannitol, sucrose, trehalose, PEG or a combination thereof, and more specifically, sucrose, trehalose, PEG or a combination thereof. The PEG may be specifically PEG400 or PEG4000, but is not particularly limited thereto. In the formulation, the polyol may be present at a concentration of 0.1-100 mg / mL.

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

  Further, the term "amino acid" in the present invention includes all analogs, solvates, hydrates, stereoisomers, and pharmaceutically acceptable salt forms of the relevant amino acid having substantially the same potency. Things.

  The “pharmaceutically acceptable salt” in the present invention includes a salt derived from a pharmaceutically acceptable inorganic acid, organic acid or base. Examples of suitable acids include hydrochloric, bromic, sulfuric, nitric, perchloric, fumaric, maleic, phosphoric, glycolic, lactic, salicylic, succinic, toluene-p-sulfonic, tartaric, acetic acid , Citric acid, methanesulfonic acid, formic acid, benzoic acid, malonic acid, naphthalene-2-sulfonic acid, benzenesulfonic acid and the like. Salts derived from appropriate bases include alkali metals such as sodium and potassium, alkaline earth metals such as magnesium, ammonium and the like.

  The “solvate” in the present invention means a compound in which an amino acid or a salt thereof forms a complex with a solvent molecule.

  More specifically, the stabilizer comprises (i) sucrose or trehalose, (ii) PEG having a number average molecular weight of 200 to 600, or PEG having a number average molecular weight of 1000 to 8000, (iii) glycine or leucine, And (iv) may be selected from the group consisting of at least two combinations of the above (i) to (iii), but are not particularly limited thereto.

  More specifically, the stabilizer includes 1) any one of sucrose, trehalose and PEG400, 2) a combination of sucrose or trehalose and glycine or leucine, 3) a combination of sucrose or trehalose and glycine and leucine, 5) a combination of sucrose or trehalose, PEG 4000 and glycine, 6) a combination of sucrose or trehalose, PEG 4000 and leucine, 7) a combination of sucrose or trehalose, PEG 4000, glycine and leucine. And 8) one selected from the group consisting of glycine, but is not particularly limited thereto.

  The surfactant contained in the liquid preparation of the present invention may be a nonionic surfactant. More specifically, the surfactant may be a polysorbate or a poloxamer.

  Specifically, the surfactant may be polysorbate 80, polysorbate 20, or poloxamer 188, but is not particularly limited thereto.

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

  Arginine contained in the liquid preparation of the present invention may exist in the form of a salt. More specifically, it may be in the form of a pharmaceutically acceptable salt.

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

  In said formulation, arginine may be present at a concentration of 0.1 to 200 mM. More specifically, arginine may be present at a concentration of 0.1 to 140 mM when the antibody is present in the formulation at 100 mg / mL, and may be present at a concentration of 0.1 to 140 mM when the antibody is present in the formulation at 50 mg / mL. It may be present at a concentration of 1 to 100 mM, but is not particularly limited thereto.

  The arginine may be included in the liquid formulation of the present invention as a viscosity reducing agent.

  By including arginine, the liquid formulation of the present invention has a viscosity of about 1 to 6 cps, but is not particularly limited thereto. The measurement of the viscosity can be performed by various methods known in the art, for example, the method of Example 1 of the present invention, but is not particularly limited thereto.

  The liquid preparation of the present invention may further contain an antioxidant.

  The “antioxidant” in the present invention plays a role in suppressing the generation of impurities that may occur in a solution state due to a protein oxidation reaction or the like.

  Such antioxidants include sodium bisulfate, ascorbic acid, ascorbyl palmitate, citric acid, butylhydroxyanisole (BHA), butylhydroxytoluene (BHT), thioglycerin, propylgallate, methionine, sodium ascorbate, citric acid Sodium acid, sodium sulfide, sodium sulfite, EDTA and other antioxidants. However, it is not limited to these.

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

  The pH of the liquid preparation of the present invention may be 4 to 6, but is not particularly limited thereto.

  On the other hand, although not particularly limited thereto, the liquid formulation may not include additional salts and / or buffers.

  For example, a preparation containing 50 mg / mL or more of the anti-TNFα antibody may not contain an additional salt and / or buffer, but is not particularly limited thereto.

  As shown in one embodiment of the invention, formulations according to the invention without them, as compared to formulations containing salts, buffers or both, provide the anti-TNFα antibody with greater stabilization against heat. However, it is not particularly limited to this.

  On the other hand, in a dosage form of a high concentration of antibody drug, eg, a drug in which 50 mg / mL or more antibody is present, an additional buffer or an amount that causes the osmotic pressure of the solution to deviate from the osmotic pressure range equivalent to the body fluid. If no additional additives are used, pain will be reduced upon administration, thus improving patient convenience.

  On the other hand, although not particularly limited thereto, the liquid preparation according to the present invention has the following effects.

  The liquid preparation according to the present invention containing arginine exhibits a relatively low content of high molecular impurities (HMW), since aggregation of the anti-TNFα antibody protein is suppressed as compared with the preparation not containing arginine, and / or Compared to a preparation containing no arginine, the production of an acidic isomer antibody is suppressed, so that a relatively low acidic isomer antibody is contained. In particular, the liquid preparations according to the present invention have the effect of reducing the production of antibodies due to denaturation and / or reducing the aggregation and particle generation against specific physical stress.

  Solution dosage forms according to the invention may further comprise a preservative. The preservative means a compound added to a pharmaceutical formulation to act as an antibacterial agent. Examples of such a preservative include benzalkonium chloride, benzethonium, chlorhexidine, phenol, m-cresol, benzyl alcohol, Examples include, but are not limited to, methyl paraben, propyl paraben, chlorobutanol, o-cresol, p-cresol, chlorocresol, phenylmercuric nitrate, thimerosal, benzoic acid, and the like. One of these preservatives may be used alone, or at least two of them may be used in appropriate combination.

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

  In addition, various pharmaceutically acceptable carriers may be included in addition to the above components.

  The dosage form of the present invention may be used for rheumatoid arthritis, psoriasis, psoriatic arthritis, axial spondyloarthritis (eg, ankylosing spondylitis, severe axial spondyloarthritis without radiologically confirmed ankylosing spondylitis), vasculitis Prevention or treatment of Alzheimer's disease, ulcerative colitis, intestinal Behcet's disease, purulent sweat glanditis, uveitis, pediatric idiopathic arthritis, pediatric psoriasis vulgaris, Crohn's disease (including adult Crohn's disease and pediatric Crohn's disease) , But is not limited thereto.

  The dosage form according to the present invention may be administered to the body by oral administration or parenteral administration including, but not limited to, subcutaneous, intramuscular, intraperitoneal, intrasternal, transdermal and intravenous injection and infusion. Not something.

  Another embodiment of the present invention is a method for producing a liquid preparation of the anti-TNFα antibody, comprising mixing an anti-TNFα antibody, a stabilizer, a surfactant, and arginine.

  The terms are as described above. Needless to say, all the specific examples of the terms also apply to the present embodiment.

  Yet another aspect of the present invention is a method for increasing the stabilization of an anti-TNFα antibody using a composition comprising a stabilizer, a surfactant and arginine.

  The terms are as described above. Needless to say, all the specific examples of the terms also apply to the present embodiment.

  Furthermore, another object of the present invention is to provide a method for increasing the stabilization of an anti-TNFα antibody using a composition that does not contain a buffer and contains a stabilizer, a surfactant, and arginine.

  The terms are as described above. Needless to say, all the specific examples of the terms also apply to the present embodiment.

  Hereinafter, the present invention will be described in detail with reference to examples. However, these examples are for the purpose of exemplifying the present invention, and the present invention is not limited to these examples.

Confirmation of Viscosity Lowering Effect and Stability of Adalimumab Solution by Additive In order to confirm the additive used for the production of adalimumab liquid preparation, sucrose 55 mg / mL, methionine 5 mM, polysorbate 80 1 mg / mL, adalimumab 100 mg / mL, pH 5. A dosage form 1 having a composition of 2 was prepared.

  Further, arginine hydrochloride, lysine hydrochloride, leucine, isoleucine, phenylalanine, glutamic acid, glycine, proline, alanine, sodium chloride, calcium chloride, and magnesium chloride were further added to the composition of dosage form 1 to form dosage forms 2 to 13, respectively. Was prepared, and the viscosity was measured using an mVROC instrument manufactured by Rheosense. Table 1 and FIG. 1 show the types and concentrations of additives added to the dosage forms and the viscosities of the dosage forms.

  As shown in the viscosity value of each dosage form in Table 1, in the dosage form 1 comprising sucrose, methionine, polysorbate 80, and adalimumab, the viscosity was 3.23. In comparison, in the dosage form to which amino acids such as leucine, isoleucine, phenylalanine, glycine, and proline were added, the viscosity was 3.20 to 3.28, and did not change much. In contrast, it was confirmed that the viscosity was reduced to 2.94 to 3.14 in the dosage form to which arginine hydrochloride, lysine hydrochloride, glutamic acid, sodium chloride, calcium chloride, magnesium chloride and the like were added.

  In order to compare the stability of each dosage form, each dosage form was sterile-filtered with a filter having a pore size of 0.2 μm, filled into a glass syringe having a capacity of about 1.0 mL by 0.4 mL, and sealed. Stored at 40 ° C. for 2 months. After storage, each sample is analyzed by SE-HPLC and contains high molecular weight impurities (HMW) such as oligomers and aggregates and low molecular weight impurities (LMW) which are fragments of adalimumab molecules. The amount was measured. First, the SE-HPLC results of the dosage forms with reduced viscosity, that is, dosage form 2, dosage form 3, dosage form 7, dosage form 11, dosage form 12, dosage form 13, and dosage form 1 as a control group are shown in Table 2. Shown in In addition, the SE-HPLC results of the dosage forms whose viscosities do not significantly change, namely, dosage form 4, dosage form 5, dosage form 6, dosage form 8, dosage form 9, dosage form 10 and dosage form 1 as a control group are shown in the table. 3 is shown.

  From the results in Table 2, the HMW, LMW and the total amount of impurities before storage were the same, but in the total amount of impurities after storage at 40 ° C. for 2 months, the dosage form 11 containing sodium chloride contained 7.10% and calcium chloride The dosage form 12 containing 7.38% and the dosage form 13 containing magnesium chloride are 7.42%, which can be significantly increased as compared with the total amount of impurities of the control dosage form 1 of 6.59%. confirmed. In contrast, dosage form 2, dosage form 3, and dosage form 7 containing arginine hydrochloride, lysine hydrochloride, and glutamic acid have a total impurity amount of 6.45 to 6.61% after storage at 40 ° C. for 2 months. , The same level as 6.59% of the total amount of impurities in the control form, dosage form 1.

  From the results in Table 3, the dosage form containing amino acids such as leucine, isoleucine, phenylalanine, glycine, and proline has a total impurity amount of 5.94 to 6.17% after storage at 40 ° C. for 2 months, and the control group It was confirmed that the content was significantly reduced as compared with 6.59% of the impurity content of Dosage Form 1 which was the following. Therefore, it was confirmed that amino acids such as leucine, isoleucine, phenylalanine, glycine, and proline contribute to the stability of adalimumab.

Stability evaluation by content of arginine hydrochloride 1
In order to evaluate the stability of adalimumab dosage form according to the content of arginine hydrochloride, a dosage form was prepared as follows.

  Dosage form 14 was prepared to be 55 mg / mL of sucrose, 5 mM of methionine, 1 mg / mL of polysorbate 80 and 100 mg / mL of adalimumab, and the composition of dosage form 14 was added with 20 mM of arginine hydrochloride and 40 mM of arginine hydrochloride. Dosage forms 15 and 16 were prepared, and 0.4 mL was filled into a 1 mL glass syringe. Each syringe was stored at 40 ° C. for 2 months, and after storage, analyzed by SE-HPLC to evaluate the stability. Table 4 shows the composition of each dosage form and the HMW content before / after storage.

  In the SE-HPLC results in Table 4, the HMW content of the sample before storage was increased when the content of arginine hydrochloride was increased from 0 mM (formulation 14) to 20 mM (formulation 15) and 40 mM (formulation 16). The HMW content after storage at 40 ° C. for 2 months was 1.00% for arginine hydrochloride-free dosage form 14, and 0.16 to 0.17% for arginine hydrochloride. When the dosage forms 15 and 16 containing 20 mM and 40 mM contain 0.82% of arginine hydrochloride and 20 mM or 40 mM of arginine hydrochloride, the degree of HMW generation is reduced as compared with those without arginine hydrochloride. Therefore, it was confirmed that arginine hydrochloride has an effect of preventing adalimumab from aggregating.

Comparison of the degree of charge variant production by arginine content To compare the degree of charge variant production by arginine content, prepare dosage forms with and without arginine and store at 40 ° C for 1 month After storage, the production aspects of charge variants were compared by CEX-HPLC. Table 5 shows each dosage form and the charge variant content before and after storage.

  Dosage form A was made to contain 55 mg / mL sucrose, 5 mM methionine, 1 mg / mL polysorbate 80 (PS80) and 100 mg / mL adalimumab, and dosage form B was prepared by adding arginine hydrochloride (Arginine hydrochloride) to the composition of dosage form A. Hydrochloride) was prepared to further contain 20 mM. The content of the acidic isomer antibody before storage at 40 ° C. for one month is the same, and when the content of the acidic isomer antibody after storage is compared, the dosage form B containing arginine hydrochloride is compared with the dosage form A. It can be seen that the content of the acidic isomer antibody is low and the content of K0 is high.

  Therefore, it was confirmed that arginine hydrochloride reduces the production of acidic isomer antibodies of adalimumab.

Comparison of Dosage Form and Dosage Form Stability by Type of Surfactant In order to compare the stability of the adalimumab liquid formulation by type of surfactant, a dosage form having the following composition was prepared.

  Similarly to the dosage form 14 of Example 2, a dosage form 17 was prepared in which sucrose was 55 mg / mL, methionine was 5 mM, polysorbate 80 was 1 mg / mL, and adalimumab was 100 mg / mL. Further, the content of the surfactant was fixed to be the same, and the types of the surfactant were changed to polysorbate 20 and poloxamer 188, to thereby prepare a dosage form 18 and a dosage form 19.

  Each dosage form was sterilized, filtered, filled into a 1 mL glass syringe in 0.4 mL portions, and stored at 40 ° C. for 1 month. Samples before and after storage were analyzed by SE-HPLC to determine the impurity content of HMW and LMW. Table 6 shows the structures of the dosage forms 17 to 19 and the results of analyzing the SE-HPLC content before and after storage at 40 ° C. for one month.

  In the results of Table 6, the contents of HMW and LMW before and after storage at 40 ° C. for 1 month did not change so much depending on the type of surfactant. That is, the stability of the dosage form containing polysorbate 80, the dosage form containing polysorbate 20, and the dosage form containing poloxamer 188 were equivalent.

  Therefore, it was confirmed that the effects of polysorbate 80, polysorbate 20, and poloxamer 188 on the stability of adalimumab were equivalent.

Comparison of dosage form and stability by type of polyol In order to compare the stability of adalimumab liquid preparation by type of polyol, a dosage form having the following composition was prepared.

  Similarly to the dosage form 14 of Example 2, a dosage form 20 was prepared so as to be 55 mg / mL of sucrose, 5 mM of methionine, 1 mg / mL of polysorbate 80, and 100 mg / mL of adalimumab (same as the dosage form 17 of Example 4). ). In addition, the total content of the polyol was fixed to be the same, and the types of the polyol were changed to trehalose, PEG400, and PEG4000 to prepare dosage forms 21 to 23. Each dosage form was sterilized, filtered, filled into a 1 mL glass syringe in 0.4 mL portions, and stored at 40 ° C. for 1 month. Samples before and after storage were analyzed by SE-HPLC to determine the impurity content of HMW and LMW.

  Table 7 shows the composition of the dosage forms 20 to 23 and the results of analyzing the SE-HPLC content before and after storage at 40 ° C. for one month.

  From the results in Table 7, it was confirmed that the content of HMW and LMW before and after storage at 40 ° C. for one month greatly changed depending on the type of polyol. All the dosage forms had the same impurity content of HMW and LMW before storage at 40 ° C. for 1 month, but after storage at 40 ° C. for 1 month, the dosage form 20 containing sucrose, trehalose and PEG400 ~ 22, the HMW content was relatively low compared to dosage form 23 containing PEG4000. All LMW contents were equivalent. Thus, it was confirmed that there was a difference in the stability of the dosage form depending on the type of the polyol used, and it was confirmed that sucrose and trehalose were more effective than other polyols in improving the stability.

Comparison of the stability of a Humira dosage form with a dosage form composed of a polyol, arginine, methionine, a surfactant, and an additional stabilizer In a composition using 5 mM of methionine as a stabilizer, the type of the surfactant or the polyol, Various compositions are prepared by changing the presence or absence of a stabilizer, the content of arginine hydrochloride, etc. to prepare samples, the compositions are prepared in a commercially available Humira dosage form, and they are stored at 40 ° C. -Stability of each dosage form was analyzed by HPLC. Table 8 shows the composition of each dosage form, and Table 9 shows the impurity content of each composition before and after storage at 40 ° C for 2 months.

* Commercially available Humira dosage forms: Sodium Phosphate Monobasic Dihydrate (0.86 mg / mL), Sodium Phosphate Dibasic Dihydrate (1.53 mg / mL), Sodium Citrate (0.3 mg / ml / cid / cid / cid / cid / cid / cid / cid / cid / cid / cid / cid / cid / cid / cid / cid / cid / cid / cid / cid / cid / cid / cid / cid / cid / cid / cid / cid.htm). ), Mannitol (12 mg / mL), Sodium Chloride (6.16 mg / mL), Polysorbate 80 (PS80) (1 mg / mL), pH 5.2

  As shown in Table 8, sucrose, trehalose, a combination of sucrose and PEG4000, and a combination of trehalose and PEG4000 were used as polyols, and polysorbate 80 and poloxamer 188 were used as surfactants. Arginine was used at 0, 20 or 40 mM, and to confirm the role of additional additives, a dosage form was designed with glycine (Gly), leucine (Leu) or a combination of glycine and leucine, at adalimumab 100 mg / mL. Stability was compared to a commercial Humira dosage form. The stability at 50 mg / mL was also compared for the compositions of the commercial Humira dosage form and dosage form 44 (identical to dosage form 26 except for the adalimumab content).

  A comparison of the stability of each dosage form in Table 9 confirms that all dosage forms show superior stability over the commercial Humira dosage form. The total amount of impurities before storage was 0.57 to 0.74, which were all the same. However, in terms of stability after storage at 40 ° C. for 2 months, a commercial Humira dosage form with a content of adalimumab of 100 mg / mL (formulation 43) and a commercial Humira with a content of adalimumab of 50 mg / mL. It was confirmed that the adalimumab impurity content of other dosage forms was lower than that of the dosage form (formulation 45).

  Therefore, it was confirmed that the present dosage form comprising various polyols, a surfactant, arginine hydrochloride, and an additional stabilizer is a more stable dosage form in terms of increasing impurities than a commercially available Humira dosage form. Was.

  Further, in Example 5, when PEG4000 was used as the polyol, the total amount of impurities after storage at 40 ° C. for one month was about 0.4% higher than that in the dosage form using sucrose and trehalose. In the examples, despite the storage period of 2 months, the use of PEG 4000 with sucrose or trehalose resulted in the same impurity content of adalimumab as compared to using sucrose and trehalose alone. Was. Therefore, replacing part of sucrose or trehalose with PEG having a high molecular weight is useful as a method for maintaining osmotic pressure and stability when using an additional additive to obtain further effects such as antioxidation. It was confirmed that.

Comparative evaluation of the stability of commercial Humira dosage forms and dosage forms containing arginine hydrochloride against mechanical stress Comparison of the stability of commercial Humira dosage forms with dosage forms containing arginine hydrochloride against mechanical stress For this purpose, a sample having the following composition was prepared and passed through a rotary piston pump to evaluate the number of particles before and after passing through the pump.

  In addition, in order to confirm whether the measured particles are derived from adalimumab, a placebo was prepared by removing only adalimumab from the composition of each sample and passed through a rotary piston pump under the same conditions before and after passing the pump. Was evaluated. The method for evaluating the number of particles is to use a micro flow imaging device manufactured by Protein Simple. Table 10 shows the composition of each sample, and Table 11 and FIG. 2 show the number of particles before and after the sample and the placebo passed through the pump.

  As shown in Table 10, the dosage form 46 and the dosage form 47 have the same additive composition, the adalimumab content is different, the dosage form 46 is 100 mg / mL, and the dosage form 47 is 50 mg / mL. is there. In dosage form 48, the composition and adalimumab content were adjusted to be the same as those of commercially available Humira. As a result of analyzing the number of particles before and after the passage by passing a rotary piston pump through each dosage form, as shown in Table 11 and FIG. 2, the dosage form 46 was 85743 particles / mL and the dosage form 47 was 53732 particles / mL. It was confirmed that the dosage form 46 containing a large amount of adalimumab had more particles than the dosage form 47 containing a small amount of adalimumab. On the other hand, in the case of the commercially available Humira dosage form 48, the number of particles after passing through the pump was 150617 / mL, which was larger than that of the dosage forms 46 and 47. In addition, the particle concentration of placebo in all dosage forms was 3618 particles / mL and 7938 particles / mL after passing through the pump, which was significantly lower than the particle concentration of the sample containing adalimumab after passing through the pump. It was confirmed that the particles measured in the sample containing were derived from adalimumab. Thus, it can be seen that the dosage form containing arginine hydrochloride effectively protects adalimumab from mechanical stress as compared to the commercially available Humira dosage form.

Experiment of Viscosity Lowering Effect by Arginine Hydrochloride Concentration The following experiment was performed to confirm the range of arginine concentration at which viscosity decreases when arginine is added to adalimumab solution. As a control group not containing arginine hydrochloride (ArgHCl), a sample was prepared so as to contain adalimumab 100 mg / mL and polysorbate 80 1 mg / mL, or adalimumab 50 mg / mL and polysorbate 80 1 mg / mL. As a test group, the concentration of arginine hydrochloride was increased by 20 mM in the composition of each control group, and a sample containing arginine hydrochloride at a final concentration of 180 mM at the maximum was prepared. The pH of all samples was about 5.2. The viscosity of the prepared sample was measured with an mVROC instrument manufactured by Rheosense. Table 12 shows the measurement results of the viscosity.

  As shown in Table 12, adalimumab 100 mg / ml, polysorbate 80 1 mg / mL solution without ArgHCl at the viscosity of adalimumab 100 mg / mL, polysorbate 80 1 mg / mL and various concentrations of arginine hydrochloride (ArgHCl) additive forms Had a viscosity of 2.71 cp. It was confirmed that when ArgHCl was added thereto at a final concentration of 20 to 120 mM, the viscosity was reduced as compared with the composition without ArgHCl. When ArgHCl was added at a final concentration of 140 mM, the viscosity was equivalent to the composition without ArgHCl added. It was confirmed that when ArgHCl was added at a final concentration of 160 mM or more, the effect of lowering the viscosity did not appear, and the viscosity increased as compared with the dosage form without ArgHCl. In the dosage form containing 50 mg / mL adalimumab and 1 mg / mL polysorbate 80, the viscosity without ArgHCl is 1.47 cp, and the viscosity when the concentration of ArgHCl is 20-80 mM is 1.42-1.45. And had a lower viscosity than the ArgHCl non-added form. The viscosity when ArgHCl was added at a final concentration of 100 mM was equivalent to the viscosity of the composition without the addition of ArgHCl. When ArgHCl was added at a final concentration of 120 mM or more, the viscosity was higher than when it was not added.

  Therefore, it was confirmed that in the 100 mg / mL adalimumab solution, the viscosity was reduced by adding 140 mM or less of ArgHCl, and in the 50 mg / mL adalimumab solution, the viscosity was reduced by adding 100 mM or less of arginine. .

Experiment 1 to confirm the effect of buffer and salt on adalimumab preparations 1
In order to understand the effects of buffers and salts on the stability of adalimumab, a sample of a dosage form containing no buffer or salt was prepared, and a sample of a dosage form containing a buffer or salt was prepared. After storage at 40 ° C for 2 weeks and 1 month, the stability was compared by SE-HPLC, and the pH of each sample was measured.

* 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 hydramine / m3d.

  Table 13 shows the dosage forms of the samples and the results of the SE-HPLC analysis of each sample after zero-time storage at 40 ° C. for 2 weeks and storage at 40 ° C. for 1 month. A-1 samples were prepared so that adalimumab was 100 mg / mL and PS80 1 mg / mL, and A-2 to A-5 were prepared so that they contained a buffer or a salt. A-6 to A-10 were prepared by adjusting the concentration of adalimumab in the compositions of A-1 to A-5 to 50 mg / mL. From the SE-HPLC results of each sample, comparing A-1 containing no buffer with A-2 containing buffer (or comparing A-6 with different adalimumab concentrations and A-7), the buffer containing The smaller dosage form showed a smaller increase in HMW and LMW, indicating a more stable dosage form.

  Also, comparing A-1 and A-6 with dosage forms containing salts such as NaCl, ammonium sulfate (Ammonium Sulfate), and sodium sulfate (Sodium Sulfate), a dosage form containing no salt is better than a dosage form containing salt. Therefore, the increase width of HMW and LMW is small. Thus, it can be seen that adalimumab dosage forms containing no salt are preferable in view of the stability of adalimumab.

  Also, the pH of all samples was 5.2 before storage, after storage for two weeks, and after storage for one month, and was kept constant. Therefore, it was confirmed that adalimumab at 50 mg / mL or more had a sufficient self-buffering effect, so that the use of a buffer was unnecessary. Further, it was confirmed that a highly stable dosage form could be constituted by not using a buffer.

Confirmation experiment of the effect of buffer and salt on adalimumab preparation 2
To understand the effects of buffers and salts on adalimumab stability, adalimumab (100 mg / mL or 50 mg / mL), stabilizing agent (sucrose 55 mg / mL or glycine 160 mM), arginine hydrochloride (ArgHCl) 50 mM, methionine A sample of a dosage form composed of 5 mM and polysorbate 80 1 mg / mL was prepared, and a sample of a dosage form in which a buffer or a salt was added to the dosage form was prepared. For comparison, a sample having a Humira composition of adalimumab 100 mg / mL or adalimumab 50 mg / mL was prepared, and each dosage form was filled into a glass syringe by 0.4 mL, stored at 55 ° C. for 1 week, and subjected to SE-HPLC. The stability was compared and the pH was measured. The composition and monomer content before / after storage at 55 ° C. for 1 week are shown in the table below.

* Humira composition: Sodium phosphate monobasic dihydrate 0.86 mg / mL, Sodium phosphate dibasic dihydrate 1.53 mg / mL, Sodium citrate / modichloride 0.3 mg / mL, citric acid. 16mg / mL, PS80 1mg / mL
** Humira Composition Buffer: Sodium phosphate monobasic dihydrate 0.86 mg / mL, Sodium phosphate dibasic dihydrate 1.53 mg / mL, Sodium citrate 0.3 mg / mlc.

  First, the pH of all samples was 5.2 before and after storage at 55 ° C. for one week and was kept constant. Therefore, it was confirmed that adalimumab of 50 mg / mL or more has a sufficient self-buffering effect in the compositions and the similar compositions in the above table.

As shown in Table 14, the monomer content of each sample before storage was 98.8 to 99.0%, which was equivalent. After storage at 55 ° C. for one week, the monomer content was a value that varied depending on the composition. The sample (A-11) having the composition of adalimumab 100 mg / mL, sucrose 55 mg / mL, ArgHCl 50 mM, methionine 5 mM and polysorbate 80 1 mg / mL has a monomer content of 94.8% after storage, NaCl, ammonium sulfate, salts of MgCl ₂ and CaCl _2, or the use of sodium acetate buffer (a-12~A-16), the content of the monomer after storage becomes 94.1 to 94.7% In addition, the monomer content was lower than that of a sample having a composition in which no salt or buffer was used. In the case of using Gly 160 mM instead of sucrose as a stabilizer (A-17), the content of the monomer after storage was 95.2%, while the use of a salt or a buffer (A-17) -18 to A-23), the monomer content after storage was 94.0 to 94.9%, and the monomer content was lower than the composition without using a salt or a buffer. Even in the composition in which the content of adalimumab was reduced to 50 mg / mL, the one using sucrose as a stabilizer without using a salt or a buffer (A-25) had a monomer content of 95.000 after storage. 2% and Glycine (Gly) as stabilizer (A-32) has a monomer content of 95.6% after storage, but additional salt / buffer in each dosage form When the agent is used, the dosage form (A-26 to A-31) having a monomer content of sucrose after storage becomes 93.5 to 95.0%, and the dosage form containing Gly (A-33) -A-38) is 93.6-95.4%, and the monomer content of the dosage form with additional salt / buffer is higher than that of the dosage form without salt / buffer. Less than body content. Therefore, it has been confirmed that unless an additional salt or buffer is used in a dosage form containing adalimumab, arginine, a stabilizer and a surfactant, the stability of adalimumab is increased, and by utilizing the buffering effect of adalimumab itself. It was confirmed that a composition having increased stability could be constituted without using an additional buffer.

  However, when compared with the Humira composition sample (A-24, A-39) having the same adalimumab content, the monomer content of the dosage form containing the salt and the buffer after storage at 55 ° C. for one week is different from that of the Humira composition. The monomer content was higher than the sample after storage at 55 ° C. for one week.

  Therefore, adalimumab dosage forms containing arginine, surfactants and stabilizers preferably do not use additional salts or buffers in terms of stability, but dosage forms containing arginine, surfactants and stabilizers are preferred. , Regardless of whether additional salts or buffers were included, were found to be more stable than the commercial Humira dosage form.

Confirmation experiment of stabilizing effect of polyol in adalimumab preparation To compare the stabilizing effect of polyol used for enhancing the in-solution stability of adalimumab, adalimumab 112 mg / mL solution, and adalimumab 112 mg / mL and each polyol A dosage form containing 42 mg / mL was prepared as follows, and repeatedly subjected to 5 cycles and 10 cycles of cold thawing at -70 ° C. and 5 ° C. and analyzed by SE-HPLC to measure the content of HMW, LMW and monomer. did.

  Table 15 shows the SE-HPLC results at each sampling point of the sample dosage form and stability test. From the above results, when cold thawing of each sample was repeated, the increase in HMW and LMW in the dosage form to which mannitol, sucrose, or trehalose was added was smaller than that in the dosage form without addition of polyol. As a result, it was confirmed that the polyol had a stabilizing effect. Comparing the stabilizing effects of each polyol type, those with the addition of sucrose or trehalose have the same HMW and LMW content as the sample before cold thawing, and the same purity even after 10 cycles of cold thawing as before cold thawing. And it was confirmed that the stabilizing effect was large. In contrast, the sample containing mannitol showed a tendency for the HMW content to increase when cold thawing was repeated, and it was confirmed that the purity was low during the cold thawing period. Therefore, it was confirmed that sucrose and trehalose were more excellent in stabilizing effect than mannitol.

Cold thawing test of adalimumab dosage form for confirmation and comparison of stabilizing effects of arginine, methionine, glycine and sucrose To confirm the effect of arginine, methionine, glycine and sucrose on stabilization during cold thawing of the stock solution, The sample was prepared as described above, and 1 mL was filled into a 5 mL polycarbonate bottle. Immediately after the preparation, cold thawing at −70 ° C. and 5 ° C. was repeated 5 times, and then analyzed by SE-HPLC. The composition of each sample and the results of SE-HPLC before and after cold thawing are shown below.

  A sample was made to be 130 mg / mL adalimumab, 1 mg / mL polysorbate 80, and a sample was made so that it contained additional stabilizers. The impurity content of each sample before cold thawing was the same for both HMW and LMW. After cold thawing, the LMWs of all samples were comparable, but the HMW content varied depending on the type and content of the additional stabilizer. Without additional stabilizer, the HMW increased to 1.76% after 5 cold thaws, whereas with 50 mM arginine hydrochloride, the HMW was 0.67% after 5 cold thaws, Significantly reduced. In a dosage form further containing methionine in 50 mM of arginine hydrochloride, those containing 5 mM of methionine did not show a further decrease in HMW, but those containing 25 mM showed that the HMW was further reduced to about 0.52%. Was done. In the case where glycine or sucrose was further added together with arginine hydrochloride and methionine, the HMW was further reduced to 0.41 to 0.43% after 5 times of cold thawing, and the dosage form containing glycine was equivalent to the dosage form containing sucrose. It was confirmed to exhibit stability. Therefore, it was confirmed that methionine, arginine, glycine, and sucrose all contribute to the stability of adalimumab. In addition, it was confirmed that an equivalent stabilizing effect can be obtained by appropriately combining a polyol or an amino acid as a stabilizer.

Comparative study of stability of glycine-containing dosage form, sucrose-containing dosage form, and commercial Humira dosage form To compare the stability of glycine-containing dosage form, sucrose-containing dosage form, and commercial Humira dosage form , Adalimumab content was 100 mg / mL or 50 mg / mL, samples were prepared to contain 50 mM arginine hydrochloride (ArgHCl), 1 mg / mL polysorbate 80 (PS80), 5 mM methionine, and additional stabilization in its composition Glycine, a combination of glycine and methionine, or sucrose was added as an agent to prepare a sample. Further, a sample was prepared with a commercially available Humira composition such that the content of adalimumab was 100 mg / mL or 50 mg / mL. 0.4 mL of each was filled in a 1 mL glass syringe, and the contents of HMW and LMW before / after storage at 40 ° C. for 4 weeks were analyzed by SE-HPLC. Each composition and the result of SE-HPLC are shown below.

  Samples with a composition comprising arginine hydrochloride, polysorbate 80, methionine, adalimumab show equivalent stability before storage at 40 ° C. for 4 weeks. On the other hand, the sample of the commercially available Humira composition had an HMW about 0.1% higher than other samples immediately after preparation. The total content of HMW and LMW after storage is 2.81 to 2.99% for adalimumab 100 mg / mL and 2.56 to 2.71 for adalimumab 50 mg / mL in all samples including arginine hydrochloride. %, Which was relatively small, but was 4.03% at 100 mg / mL adalimumab and 4.06% at 50 mg / mL adalimumab in the commercially available Humira composition sample. And LMW content was high. Therefore, the combination of the additives of this example, that is, the dosage form containing arginine, the dosage form containing arginine and a polyol or an amino acid as an additional stabilizer is a dosage form superior in terms of stability to the commercially available Humira dosage form. Was confirmed.

Comparative study of the stability of adalimumab dosage form according to the concentration of adalimumab, sucrose, glycine, leucine, methionine, sodium chloride and arginine. In order to compare the stability of adalimumab dosage form, adalimumab, arginine hydrochloride (ArgHCl), sodium chloride ( NaCl), polysorbate 80 (PS80), methionine (Met), sucrose (Sucrose), glycine (Gly), and leucine (Leu) were used to prepare samples having various compositions. For comparison, a sample was prepared with a commercially available Humira composition such that the content of adalimumab was 100 mg / mL or 50 mg / mL. 0.4 mL of each dosage form was filled into a 1 mL glass syringe, and the monomer content before / after storage at 40 ° C. for 4 weeks was analyzed by SE-HPLC. Each composition and the result of SE-HPLC are shown below.

* Commercially available humira dosage forms: Sodium phosphate monobasic dihydrate 0.86 mg / mL, Sodium phosphate dibasic dihydrate 1.53 mg / mL, Sodium citrate 0.3 mg / ml, nicotine / demochloride chloride 6.16mg / mL, PS80 1mg / mL

  The monomer content of all samples before storage at 40 ° C. was 99.27% to 99.33%, which was comparable regardless of dosage form.

  The monomer content of the sample (A-40 to A-61) having a content of adalimumab of 100 mg / mL after storage at 40 ° C. for 4 weeks is determined by the dosage form (A-40 to A-A) containing no sodium chloride. -55) from 97.04% to 97.23%, and from 96.85% to 97.00 in dosage forms containing sodium chloride (A-56 to A-61, excluding the commercial Humira composition A-62). %, Slightly less than the dosage form without sodium chloride. However, the difference in the monomer content after storage at 40 ° C. for 4 weeks due to the content of sodium chloride, sucrose, methionine, glycine, and leucine is relatively small, and the composition containing arginine (A-40 to A- It was confirmed that the monomer content of 61) was 1% or more higher than the 95.64% monomer content of a commercially available Humira composition (A-62) having the same adalimumab concentration.

  Even when the adalimumab content was 50 mg / mL, the results were the same as the analysis results of the adalimumab composition of 100 mg / mL. The monomer content of the sample (A-63 to A-85) having a content of adalimumab of 50 mg / mL after storage at 40 ° C. for 4 weeks was determined in the dosage form containing no sodium chloride (A-63 to A-85). -79) and 97.15% to 97.38%, and 96.81% to 97.06 in dosage forms containing sodium chloride (A-80 to A-85, excluding the commercially available Humira composition A-86). %, Slightly less than the dosage form without sodium chloride. However, the difference in the monomer content after storage at 40 ° C. for 4 weeks due to the content of sodium chloride, sucrose, methionine, glycine, and leucine is relatively small, and the composition containing arginine (A-63 to A- It was confirmed that the monomer content of 85) was 1% or more higher than the monomer content of 95.47% of the commercially available Humira composition (A-86) having the same adalimumab concentration.

  Therefore, it was confirmed that the combination of additives and the similar dosage form of this example, that is, the dosage form containing arginine is a dosage form superior in stability to the commercially available Humira dosage form.

  From the above description, those skilled in the art to which the present invention pertains will appreciate that the present invention can be embodied in other specific forms without altering the technical spirit or essential characteristics. . It should be understood that the above embodiments are merely illustrative and not restrictive. The present invention should be construed to include all modifications and modified forms derived from the meaning and scope of the claims and not the specification, and equivalents thereof.

Claims (25)

  A liquid preparation of an anti-TNFα antibody, comprising an anti-TNFα antibody, a stabilizer, a surfactant, and arginine.   The liquid preparation of an anti-TNFα antibody according to claim 1, wherein the stabilizer is a polyol, another amino acid other than arginine, or a combination thereof.   The liquid preparation of an anti-TNFα antibody according to claim 2, wherein the polyol is selected from the group consisting of sucrose, trehalose, PEG and a combination thereof.   The liquid preparation of an anti-TNFα antibody according to claim 2, wherein the amino acid is selected from the group consisting of glycine, leucine, isoleucine, phenylalanine, proline and a combination thereof. The stabilizer is
(I) sucrose or trehalose,
(Ii) PEG having a number average molecular weight of 200 to 600, or PEG having a number average molecular weight of 1000 to 8000,
The liquid preparation of the anti-TNFα antibody according to claim 1, which is selected from the group consisting of (iii) glycine or leucine, and (iv) a combination of at least two of the above (i) to (iii).   The liquid preparation of an anti-TNFα antibody according to claim 2, wherein the polyol is present at a concentration of 0.1 to 100 mg / mL in the preparation.   The liquid preparation of the anti-TNFα antibody according to claim 2, wherein the amino acid other than arginine is present at a concentration of 1 to 300 mM in the preparation.   The liquid preparation of an anti-TNFα antibody according to claim 1, wherein the surfactant is a nonionic surfactant.   The liquid preparation of an anti-TNFα antibody according to claim 1, wherein the surfactant is polysorbate or poloxamer.   The liquid preparation of an anti-TNFα antibody according to claim 9, wherein the surfactant is polysorbate 80, polysorbate 20, or poloxamer 188.   The liquid formulation of an anti-TNFα antibody according to claim 1, wherein the surfactant is present at a concentration of 0.1 to 5 mg / mL in the formulation.   The liquid preparation of an anti-TNFα antibody according to claim 1, wherein the arginine is in the form of a salt.   The liquid preparation of an anti-TNFα antibody according to claim 12, wherein the arginine is in the form of arginine hydrochloride.   The liquid formulation of the anti-TNFα antibody according to claim 1, wherein arginine is present at a concentration of 0.1 to 200 mM in the formulation.   The liquid preparation of an anti-TNFα antibody according to claim 1, wherein the anti-TNFα antibody is adalimumab.   The liquid formulation of an anti-TNFα antibody according to claim 1, wherein the anti-TNFα antibody is present at a concentration of 1 to 250 mg / mL in the formulation.   The liquid formulation of an anti-TNFα antibody according to claim 1, wherein the anti-TNFα antibody is present at a concentration of 50 to 200 mg / mL in the formulation.   The liquid preparation of an anti-TNFα antibody according to claim 1, wherein the liquid preparation further comprises an antioxidant.   19. The liquid preparation of an anti-TNFα antibody according to claim 18, wherein the antioxidant is methionine.   20. The liquid formulation of an anti-TNFα antibody according to claim 19, wherein methionine is present at a concentration of 1 to 50 mM in said formulation.   21. The liquid preparation of an anti-TNFα antibody according to any one of claims 1 to 20, wherein the liquid preparation does not contain additional salts or buffers.   The liquid formulation of the anti-TNFα antibody according to any one of claims 1 to 20, wherein the pH of the liquid formulation is 4 to 6. The liquid formulation,
An anti-TNFα antibody at a concentration of 1-250 mg / mL;
A polyol having a concentration of 0.1 to 100 mg / mL;
A surfactant at a concentration of 0.1-5 mg / mL;
The liquid preparation of the anti-TNFα antibody according to claim 1, comprising arginine at a concentration of 0.1 to 200 mM. The liquid formulation,
An anti-TNFα antibody at a concentration of 1-250 mg / mL;
Other amino acids other than arginine at a concentration of 1 to 300 mM;
A surfactant at a concentration of 0.1-5 mg / mL;
The liquid preparation of the anti-TNFα antibody according to claim 1, comprising arginine at a concentration of 0.1 to 200 mM.   The method for producing a liquid preparation according to claim 1, comprising a step of mixing an anti-TNFα antibody, a stabilizer, a surfactant, and arginine.