TW201836637A - Liquid pharmaceutical agent comprising antibody characterized by realizing the stability of dimer formation inhibition and deamidation inhibition during long-term storage and being suitable for a wide range of antibody concentration - Google Patents

Abstract

Provided is a recombinant monoclonal antibody-containing liquid pharmaceutical agent which can realize the stability of dimer formation inhibition and deamidation inhibition during long-term storage and is suitable for a wide range of antibody concentration. A recombination monoclonal antibody-containing liquid pharmaceutical agent comprises 10 mg/ml ~ 200 mg/ml recombinant monoclonal antibody and 40 mM ~ 94 mM amino acid components having histidine component of less than 5 mM and has a pH value of 5.5 to 7.0. Furthermore, the amino acid components other than the histidine component have at least one component selected from the group consisting of arginine, arginine hydrochloride and methionine.

Description

 Liquid preparation containing antibody  

The present invention relates to a liquid preparation containing an antibody.

A recombinant monoclonal antibody such as Tocilizumab is currently used as an active ingredient of a pharmaceutical product. In general, in pharmaceutical pharmaceutical preparations, many proteins are highly concentrated compared to other biopharmaceutical preparations, and the concern of forming aggregates is necessary. In addition, in order to obtain a sufficient therapeutic effect, the amount of the recombinant monoclonal antibody administered per time may be large. However, for example, when a subcutaneous injection preparation capable of self-injection is used, the amount of the injection once is limited. Therefore, it may be necessary to increase the concentration of the recombinant monoclonal antibody contained in the preparation.

Further, in general, injections are known to have various administration routes such as subcutaneous administration or intravenous administration, and liquid preparations are required because of the convenience of being usable on any preparation, if only ordinary When the prescription of the lyophilized preparation used is simply dissolved, the stability which can be used as a pharmaceutical for a long period of time is not satisfied.

Heretofore, as a liquid preparation containing a high concentration of recombinant monoclonal antibody, a formulation containing a specific amount of arginine, methionine, or polysorbate is known (Patent Document 1); or a specific amount of essence is contained. A formulation of an amine hydrochloride, a histidine, or a polysorbate (Patent Document 2).

In addition, when the recombinant monoclonal antibody content is 150 mg/mL or more, the content of the amino acid component such as arginine is 100 mM or more.

 [Previous Technical Literature]    [Patent Document]  

[Patent Document 1] International Publication No. 2009/084659

[Patent Document 2] International Publication No. 2004/091658

When a liquid preparation containing a recombinant monoclonal antibody is provided, it can be used as a practical stability of a pharmaceutical product. Moreover, it is required to manufacture a liquid preparation containing a monoclonal antibody at a low cost. For example, when a preparation for subcutaneous injection, a preparation for intravenous injection, or the like, and a liquid preparation containing recombinant antibodies of different concentrations, it is required to use an aqueous solution of the same composition.

Further, conventionally, in order to prepare a liquid preparation containing a high concentration of a recombinant monoclonal antibody, it is considered necessary to add a high concentration of an amino acid component of 100 mM or more. Therefore, in order to provide a liquid preparation containing a recombinant monoclonal antibody that achieves at least one of inhibition of dimer formation, inhibition of production of decomposition products, inhibition of reduction of biological activity, and inhibition of deamidation during long-term storage, further techniques are required. Development.

The present invention provides a recombinant monoclonal antibody liquid preparation which achieves stability of at least one of dimer formation inhibition, decomposition product formation inhibition, bioactivity reduction, and deamination inhibition during long-term storage. Or a recombinant recombinant monoclonal antibody liquid preparation which achieves at least one of dimer formation inhibition, decomposition product formation inhibition, bioactivity reduction, and deamination inhibition during long-term storage, and provides a preparation at a low cost. Or a high-concentration recombinant monoclonal antibody liquid preparation that achieves at least one of dimer formation inhibition, decomposition product formation inhibition, bioactivity reduction, and deamination inhibition during long-term storage, and is provided at a low cost. A liquid preparation containing a high concentration of recombinant monoclonal antibodies capable of subcutaneous injection of kinetic viscosity. Or a recombinant recombinant monoclonal antibody liquid preparation which achieves at least one of dimer formation inhibition, decomposition product formation inhibition, bioactivity reduction, and deamination inhibition during long-term storage, and is practical at a low cost. The preparation for intravenous injection and the preparation for subcutaneous injection. Or a method for producing a recombinant recombinant monoclonal antibody liquid preparation which achieves at least one of stability of dimer formation, inhibition of decomposition product formation, reduction of biological activity, and inhibition of deamidation during long-term storage. Or a method for stabilizing a recombinant monoclonal antibody liquid preparation which achieves at least one of inhibition of dimer formation, inhibition of production of decomposition products, reduction in biological activity, and inhibition of deamidation during long-term storage. Or an aqueous solution for formulating a preparation containing a recombinant monoclonal antibody.

Further, an object of the present invention is to provide a preparation containing a recombinant monoclonal antibody which improves at least one of these, a method for producing the same, a method for the stabilization, and an aqueous solution for producing a preparation containing a recombinant monoclonal antibody.

The invention is as follows.

That is, the first aspect of the present invention is the following liquid preparation containing a recombinant monoclonal antibody.

<1-1> A liquid preparation containing a recombinant monoclonal antibody, which comprises a recombinant monoclonal antibody of 10 mg/mL to 200 mg/mL, a 45 mM to 94 mM amino acid component having a histidine component of less than 5 mM, and a pH It is 5.5~7.0.

<1-2> A liquid preparation containing a recombinant monoclonal antibody, which comprises a recombinant monoclonal antibody of 10 mg/mL to 200 mg/mL, an amino acid component of 45 mM to 94 mM having a histidine component of less than 5 mM, and a pH. It is 5.5~6.7.

<1-3> A liquid preparation such as <1-1> or <1-2>, wherein the amino acid component other than the histidine component is selected from the group consisting of arginine, arginine hydrochloride and methylamine At least one of the groups of acids.

The liquid preparation according to any one of <1-1> to <1-3> wherein the histidine component is at least selected from the group consisting of histidine and histidine hydrochloride. 1 species.

The liquid preparation according to any one of <1-1> to <1-4> which contains an amino acid component of 75 mM to 93 mM.

<1-6> The liquid preparation according to any one of <1-1> to <1-5> which contains a 90 mM amino acid component.

The liquid preparation according to any one of <1-1> to <1-6, wherein the pH is 5.8 to 6.4.

The liquid preparation according to any one of <1-1> to <1-7, wherein the pH is 6.0 to 6.2.

<1-9> The liquid preparation according to any one of <1-1> to <1-8, which contains a recombinant monoclonal antibody of 150 mg/mL to 200 mg/mL.

The liquid preparation according to any one of <1-1> to <1-9, which contains a recombinant monoclonal antibody of 170 mg/mL to 190 mg/mL.

<1-11> The recombinant monoclonal antibody liquid preparation according to any one of <1-1> to <1-10> which contains a recombinant monoclonal antibody of 180 mg/mL.

The liquid preparation according to any one of <1-1> to <1-11, which further contains a polyhydric alcohol.

The liquid preparation according to any one of <1-1> to <1-12> which further contains a surfactant.

<1-14> The liquid preparation according to <1-13>, wherein the aforementioned surfactant is a nonionic surfactant.

<1-15> The liquid preparation according to <1-14>, wherein the aforementioned nonionic surfactant is polysorbate or polyoxyethylene polyoxypropylene diol.

<1-16> The liquid preparation according to <1-15>, wherein the polysorbate is at least one selected from the group consisting of polysorbate 80, polysorbate 40, and polysorbate 20. .

<1-17> The liquid preparation according to <1-15>, wherein the polyoxyethylene polyoxypropylene diol is a polyoxyethylene (160) polyoxypropylene (30) diol.

The liquid preparation according to any one of <1> to <1-17, wherein the recombinant monoclonal antibody is selected from recombinant individual plants derived from animals (human, mouse, rat, etc.) At least one of a group of an antibody, a chimeric antibody, a humanized antibody, an antibody fragment, and a low molecular weight antibody.

The liquid preparation according to any one of <1> to <1-18, wherein the immunoglobulin type of the recombinant monoclonal antibody is selected from the group consisting of IgG (IgG1, IgG2, IgG3, IgG4), At least one of the groups of IgA, IgD, IgE, and IgM.

The liquid preparation according to any one of <1> to <1-19, wherein the immunoglobulin type of the recombinant monoclonal antibody is IgG1 (humanized IgG1 or fully human IgG1) derived from human. .

The liquid preparation according to any one of <1> to <1-20, wherein the recombinant monoclonal antibody is selected from the group consisting of an anti-tumor necrosis factor (TNF) antibody (for example, an anti-TNFα antibody), and an anti-tumor antibody. Interleukin (IL) receptor antibodies (eg, anti-IL-6 receptor antibodies, anti-IL-17 receptor antibodies), anti-IL antibodies (eg, anti-IL-5 antibodies, anti-IL-17 antibodies, anti-IL-17A antibodies) , anti-IL-1β antibody, anti-IL12/IL23-p40 antibody), anti-surface antigen antibody (eg anti-CD3 antibody, anti-CD20 antibody, anti-CD25 antibody, anti-CD30 antibody, anti-CD33 antibody, anti-CD52 antibody, anti-RANKL antibody, Anti-SLAMF7 antibody, anti-CTLA-4 antibody, anti-VEGFR-2 antibody, anti-CCR4 antibody, anti-PD-1 antibody), anti-viral antibody (such as anti-RS virus antibody), anti-integrin antibody (such as anti-α4 integrin) An antibody, an anti-vascular endothelial cell proliferation factor antibody (eg, an anti-VEGF antibody), an anti-receptor type tyrosine kinase antibody (eg, an anti-EGFR antibody, an anti-HER2 antibody), an anti-PCSK9 antibody, an anti-Dabigatran antibody, an anti-IgE antibody, and At least one of the anti-complement C5 antibodies.

The liquid preparation according to any one of <1-1> to <1-21, wherein the recombinant monoclonal antibody is adapted to be a disease selected from the group consisting of rheumatoid arthritis, juvenile idiopathic arthritis, Castleman's disease, ankylosing spondylitis, Crohn's disease, ulcerative colitis, pancreatitis, vasculitis, Kawasaki disease, systemic lupus erythematosus, cognac, dry arthritis, Sjogren Disease, Still's disease, multiple sclerosis, osteoporosis, bone disease, thrombosis, cancer (eg breast cancer, leukemia, ovarian cancer, melanoma, prostate cancer, pancreatic cancer, lymph Tumor, lung cancer, gastric cancer, renal cell carcinoma, colorectal cancer, mesothelioma, soft tissue sarcoma, multiple myeloma, etc.), cachexia, chronic rejection of transplanted organs and cells, heart failure, ischemic-induced severe arrhythmia, At least a group of hypercholesterolemia, viral infection (such as RS virus infection, HIV infection, EBV infection, etc.), plasma cell hyperplasia, hyperimmune globulinemia, anemia, renal cell hyperplasia nephritis, and asthma 1 person.

The liquid preparation according to any one of <1-1> to <1-22, wherein the recombinant monoclonal antibody is selected from the group consisting of Tocilizumab, Trastuzumab, Rituximab, Palivizumab, Infliximab, Basiliximap, Gemtuzumab ozogamicin, Bevacizumab , Ibritumomab tiuxetan, Adalimumab, Cetuximab, Ranibizumab, Omalizumab, Eculizumab, Panitumumab, Ustekinumab, Golimumab, Canakinumab, Denosumab, Mogamulizumab, Ofatumumab, Pertuzumab, Trastuzumab emtansine, Brentuximab vedotin, Natalizumab, Nivolumab, Alemtuzumab, Secukinumab, Ramucirumab and Ipilimumab At least one of the groups.

The liquid preparation according to any one of <1> to <1>, wherein the liquid preparation is a preparation for subcutaneous injection or a preparation for intravenous injection.

(1) The liquid preparation according to any one of the above <1-1> to <1-24>, such as <1- 22> The treatment of at least one of the applicable diseases described is the amount necessary for the necessary patient to be treated.

The second aspect of the present invention is the following liquid preparation containing a recombinant monoclonal antibody.

<2-1> A liquid preparation containing a recombinant monoclonal antibody, which comprises a recombinant monoclonal antibody of 10 mg/mL to 200 mg/mL, an amino acid component of 45 mM to 94 mM having a histidine component of less than 5 mM, and a pH. It is 5.5~7.0.

<2-2> A liquid preparation containing a recombinant monoclonal antibody, which comprises a recombinant monoclonal antibody of 10 mg/mL to 200 mg/mL, an amino acid component of 45 mM to 94 mM having a histidine component of less than 5 mM, and a pH It is 5.5~6.7.

<2-3> A liquid preparation such as <2-1> or <2-2>, wherein the amino acid component other than the histidine component is selected from the group consisting of arginine, arginine hydrochloride and methylamine At least one of the groups of acids.

The liquid preparation according to any one of <2-1>, wherein the histidine component is at least selected from the group consisting of histidine and histidine hydrochloride. 1 species.

<2-5> The liquid preparation according to any one of <2-1> to <2-4> which contains an amino acid component of 75 mM to 93 mM.

<2-6> The liquid preparation according to any one of <2-1> to <2-5> which contains a 90 mM amino acid component.

<2-7> The liquid preparation according to any one of <2-1> to <2-6, wherein pH is 5.8 to 6.7.

<2-8> The liquid preparation according to any one of <2-1> to <2-7, wherein the pH is 6.0 to 6.5.

<2-9> The liquid preparation according to any one of <2-1> to <2-8, which contains a recombinant monoclonal antibody of 10 mg/mL to 30 mg/mL.

<2-10> The liquid preparation according to any one of <2-1> to <2-9, which contains 20 mg/mL of recombinant monoclonal antibody.

<2-11> The liquid preparation according to any one of <2-1> to <2-8, which contains a recombinant monoclonal antibody of more than 30 mg/mL to less than 150 mg/mL.

The liquid preparation according to any one of <2-1> to <2-11> which further contains a polyhydric alcohol.

The liquid preparation according to any one of <2-1> to <2-12> which further contains a surfactant.

<2-14> The liquid preparation according to <2-13>, wherein the aforementioned surfactant is a nonionic surfactant.

<2-15> The liquid preparation according to <2-14>, wherein the aforementioned nonionic surfactant is polysorbate or polyoxyethylene polyoxypropylene diol.

<2-16> The liquid preparation according to <2-15>, wherein the polysorbate is at least one selected from the group consisting of polysorbate 80, polysorbate 40, and polysorbate 20. .

<2-17> The above polyoxyethylene polyoxypropylene diol is a liquid preparation of polyoxyethylene (160) polyoxypropylene (30) diol <2-15>.

The liquid preparation of any one of <2> to <2-17>, wherein the immunoglobulin type of the recombinant monoclonal antibody is IgG1 derived from human.

<2-19> The liquid preparation according to <2-18>, wherein the aforementioned IgG1 derived from human is Tocilizumab.

The liquid preparation according to any one of <2> to <2>, wherein the liquid preparation is a preparation for subcutaneous injection or a preparation for intravenous injection.

<2-21> A liquid preparation containing a recombinant monoclonal antibody, which comprises a recombinant monoclonal antibody of 10 mg/mL to 200 mg/mL, and an amino acid component of 45 mM to 94 mM having a histidine component of less than 5 mM, histamine The amino acid component other than the acid component is at least one selected from the group consisting of arginine, arginine hydrochloride, and methionine; and contains 0.05 to 20 mg/mL of a surfactant, pH. It is 5.5~7.0.

<2-22> The liquid preparation according to <2-21>, wherein the aforementioned surfactant is a nonionic surfactant.

<2-23> The liquid preparation according to <2-22>, wherein the aforementioned nonionic surfactant is polysorbate or polyoxyethylene polyoxypropylene diol.

<2-24> The liquid preparation according to <2-23>, wherein the polysorbate is at least one selected from the group consisting of polysorbate 80, polysorbate 40, and polysorbate 20. .

<2-25> The liquid preparation according to <2-23>, wherein the polyoxyethylene polyoxypropylene diol is a polyoxyethylene (160) polyoxypropylene (30) diol.

The liquid preparation of any one of <2-21> to <2-25>, wherein the immunoglobulin type of the recombinant monoclonal antibody is IgG1 derived from human.

<2-27> The liquid preparation according to <2-26>, wherein the aforementioned IgG1 derived from human is Tocilizumab.

<2-28> The liquid preparation according to any one of <2-21> to <2-27> which contains the recombinant monoclonal antibody of 20 mg/mL.

<2-29> A liquid preparation containing a recombinant monoclonal antibody for intravenous injection, comprising Tocilizumab of 10 mg/mL to 30 mg/mL, and amino acid component of 45 mM to 94 mM having a histidine component of less than 5 mM, group The amino acid component other than the amino acid component contains at least one selected from the group consisting of arginine, arginine hydrochloride, and methionine; and contains 0.05 to 20 mg/mL of polysorbate. 80, pH is 5.5~7.0.

<2-30> A liquid preparation according to <2-21> to <2-29> which contains 0.1 to 0.5 mg/mL of polysorbate 80.

The liquid preparation according to any one of <2-21> to <2-30> which contains an amino acid component of 75 mM to 93 mM.

<2-32> The liquid preparation according to any one of <2-21> to <2-31, which contains Tocilizumab of 10 mg/mL - 30 mg/mL, and 85 mM - 92 mM of histidine component which is less than 5 mM. The amino acid component and the amino acid component other than the histidine component contain at least one selected from the group consisting of arginine, arginine hydrochloride and methionine; and 0.15 to 0.25 mg /mL polysorbate 80, pH 5.5 ~ 7.0, and for intravenous injection.

<2-33> The liquid preparation according to any one of <2-21> to <2-32, which contains 20 mg/mL of Tocilizumab.

The third aspect of the present invention is a pharmaceutically stable preparation of the following recombinant monoclonal antibodies.

<3-1> A pharmaceutical stable preparation of a recombinant monoclonal antibody, which is a recombinant monoclonal antibody of 10 mg/mL to 200 mg/mL, an amino acid component of 45 mM to 94 mM, and a histidine component of less than 5 mM. It is composed of an aqueous solution having a pH of 5.5 to 7.0.

<3-2> A pharmaceutical stable preparation of a recombinant monoclonal antibody, which is a recombinant monoclonal antibody of 10 mg/mL to 200 mg/mL, an amino acid component of 45 mM to 94 mM, and a histidine component of less than 5 mM. It is composed of an aqueous solution having a pH of 5.5 to 6.7.

<3-3> The preparation of <3-1> or <3-2>, wherein the amino acid component other than the histidine component is selected from the group consisting of arginine, arginine hydrochloride and methionine At least one of the groups formed.

The preparation according to any one of <3-1> to <3-3, wherein the histidine component is at least 1 selected from the group consisting of histidine and histamine hydrochloride. Kind.

<3-5> The preparation according to any one of <3-1> to <3-4> which contains an amino acid component of 75 mM to 93 mM.

<3-6> The preparation according to any one of <3-1> to <3-5> wherein the pH is 5.8 to 6.7.

<3-7> The preparation according to any one of <3-1> to <3-6> which further contains a polyhydric alcohol.

<3-8> The preparation of any one of <3-1> to <3-7> which further contains a surfactant.

<3-9> The preparation according to <3-8>, wherein the aforementioned surfactant is a nonionic surfactant.

<3-10> The preparation according to <3-9>, wherein the aforementioned nonionic surfactant is polysorbate or polyoxyethylene polyoxypropylene diol.

<3-11> The preparation according to <3-10>, wherein the polysorbate is at least one selected from the group consisting of polysorbate 80, polysorbate 40, and polysorbate 20.

<3-12> The preparation according to <3-10>, wherein the polyoxyethylene polyoxypropylene diol is a polyoxyethylene (160) polyoxypropylene (30) diol.

<3-13> The preparation according to any one of <3-1> to <3-12, wherein the immunoglobulin type of the recombinant monoclonal antibody is IgG1 derived from human.

<3-14> The preparation according to <3-13>, wherein the aforementioned IgG1 derived from human is Tocilizumab.

<3-15> The aforementioned preparation is a preparation for subcutaneous injection or a preparation for intravenous injection <3-1> to <3-14>.

The fourth aspect of the present invention is the production method of the following liquid preparation containing a recombinant monoclonal antibody.

<4-1> A method for producing a liquid preparation containing a recombinant monoclonal antibody having a pH of 5.5 to 7.0 and containing 10 mg/mL to 200 mg/mL, which comprises an amino acid having a histidine component of less than 5 mM of 45 mM to 94 mM The steps of the ingredients.

<4-2> A method for producing a liquid preparation containing a recombinant monoclonal antibody of 10 mg/mL to 200 mg/mL, which has a pH of 5.5 to 6.7, which comprises an amino acid having a histidine component of less than 5 mM of 45 mM to 94 mM The steps of the ingredients.

<4-3> The production method of <4-1> or <4-2>, wherein the amino acid component other than the histidine component is selected from the group consisting of arginine, arginine hydrochloride and methylamine At least one of the groups of acids.

The method of any one of <4> to <4-3> wherein the histidine component is at least selected from the group consisting of histidine and histamine hydrochloride. 1 species.

<4-5> The production method according to any one of <4-1> to <4-4> which contains an amino acid component of 75 mM to 93 mM.

<4-6> The production method according to any one of <4-1> to <4-5, wherein the pH is 5.8 to 6.7.

<4-7> The production method according to any one of <4-1> to <4-6, which comprises a recombinant monoclonal antibody of 150 mg/mL to 200 mg/mL.

<4-8> The production method according to any one of <4-1> to <4-6, which contains a recombinant monoclonal antibody of 10 mg/mL to 30 mg/mL.

The method of any one of <4> to <4-8> which further contains a polyhydric alcohol.

The method of any one of <4> to <4-9> which further contains a surfactant.

<4-11> The manufacturing method of <4-10> wherein the surfactant is a nonionic surfactant.

<4-12> The production method according to <4-11>, wherein the nonionic surfactant is polysorbate or polyoxyethylene polyoxypropylene diol.

<4-13> The production method of <4-12>, wherein the polysorbate is at least one selected from the group consisting of polysorbate 80, polysorbate 40, and polysorbate 20. .

<4-14> The production method according to <4-12>, wherein the polyoxyethylene polyoxypropylene diol is a polyoxyethylene (160) polyoxypropylene (30) diol.

<4-15> The production method according to any one of <4-1> to <4-14, wherein the immunoglobulin type of the recombinant monoclonal antibody is IgG1 derived from human.

<4-16> The production method according to <4-15>, wherein the aforementioned IgG1 derived from human is Tocilizumab.

<4-17> The production method according to any one of <4-1> to <4-16, wherein the pH of the step of adding a 45 mM to 94 mM amino acid component having a histidine component of less than 5 mM is A method for producing a liquid preparation containing a recombinant monoclonal antibody of 10 mg/mL to 200 mg/mL from 5.5 to 7.0, which comprises the following steps A to C:

Step A: Step of preparing an antibody drug substance comprising any of the following steps A-1 to A-3

Step A-1: Step of melting the frozen antibody drug substance into a liquid state

Step A-2: Steps of preparing a liquid antibody drug substance containing any solvent

Step A-3: Steps of preparing the antibody raw material of the powder

Step B: adding an amino acid to the solvent to prepare an additive solution

Step C: a step of mixing the drug substance prepared in the step A with the additive solution prepared in the step B.

<4-18> The production method according to <4-17>, which further comprises the following step D: Step D: a step of bringing the pH of the solution prepared in the step C to 5.5 to 7.0.

<4-19> The production method according to any one of <4-1> to <4-16, wherein the pH of the step of adding a 45 mM to 94 mM amino acid component having a histidine component of less than 5 mM is A method for producing a liquid preparation containing recombinant monoclonal antibodies of 10 mg/mL to 200 mg/mL from 5.5 to 6.7, which comprises the following steps A to C:

Step A: Step of preparing an antibody drug substance comprising any of the following steps A-1 to A-3

Step A-1: Step of melting the frozen antibody drug substance into a liquid state

Step A-2: Steps of preparing a liquid antibody drug substance containing any solvent

Step A-3: Steps of preparing the antibody raw material of the powder

Step B: adding an amino acid to the solvent to prepare an additive solution

Step C: a step of mixing the drug substance melted in the step A and the additive solution prepared in the step B.

<4-20> The production method according to <4-19>, which further comprises the following step D: Step D: a step of bringing the pH of the solution prepared in the step C to 5.5 to 6.7.

<4-21> The production method according to any one of <4-1> to <4-20, which further comprises the following step E: Step E: a step of sterilizing the prepared solution by filtration.

<4-22> The production method according to <4-21>, which further comprises the following step F: Step F: a step of filling the solution prepared in the step E and capping.

The fifth aspect of the present invention is the stabilization method of the following solution containing a recombinant monoclonal antibody.

<5-1> A method for stabilizing a solution of a recombinant monoclonal antibody having a pH of 5.5 to 7.0 and containing 10 mg/mL to 200 mg/mL, which comprises an amino acid having a histidine component of less than 5 mM of 45 mM to 94 mM. The steps of the ingredients.

<5-2> A method for the stabilization of a solution containing a recombinant monoclonal antibody of 10 mg/mL to 200 mg/mL, which has a pH of 5.5 to 6.7, which comprises an amino acid having a histidine component of less than 5 mM of 45 mM to 94 mM. The steps of the ingredients.

<5-3> The stabilization method of <5-1> or <5-2>, wherein the amino acid component other than the histidine component is selected from the group consisting of arginine, arginine hydrochloride and methyl sulfide At least one group of amine acids.

<5-4> The method for stabilization according to any one of <5-1> to <5-3, wherein the histidine component is selected from the group consisting of histidine and histamine hydrochloride. At least one.

<5-5> The method of stabilization according to any one of <5-1> to <5-4> which contains an amino acid component of 75 mM to 93 mM.

<5-6> The method of stabilization according to any one of <5-1> to <5-5>, wherein the pH is 5.8 to 6.4.

<5-7> The method for stabilization according to any one of <5-1> to <5-6>, which comprises a recombinant monoclonal antibody of 150 mg/mL to 200 mg/mL.

<5-8> The method for stabilization according to any one of <5-1> to <5-6>, which comprises a recombinant monoclonal antibody of 10 mg/mL to 30 mg/mL.

<5-9> The method for stabilization according to any one of <5-1> to <5-6>, which comprises a recombinant monoclonal antibody of more than 30 mg/mL to less than 150 mg/mL.

<5-10> The method of stabilization according to any one of <5-1> to <5-9, which further contains a polyhydric alcohol.

<5-11> The method of stabilization according to any one of <5-1> to <5-10> which further contains a surfactant.

<5-12> The method for stabilization according to <5-11>, wherein the surfactant is a nonionic surfactant.

<5-13> The method for stabilization according to <5-12>, wherein the nonionic surfactant is a polysorbate or a polyoxyethylene polyoxypropylene diol.

<5-14> The method for stabilization according to <5-13>, wherein the polysorbate is at least 1 selected from the group consisting of polysorbate 80, polysorbate 40, and polysorbate 20. Kind.

<5-15> The method for stabilization according to <5-13>, wherein the polyoxyethylene polyoxypropylene diol is a polyoxyethylene (160) polyoxypropylene (30) diol.

<5-16> The method for stabilization according to any one of <5-1> to <5-15, wherein the immunoglobulin type of the recombinant monoclonal antibody is IgG1 derived from human.

<5-17> The method for stabilization according to <5-16>, wherein the aforementioned IgG1 derived from human is Tocilizumab.

The sixth aspect of the present invention is an aqueous solution for producing a liquid preparation containing a recombinant monoclonal antibody.

<6-1> An aqueous solution for producing a liquid preparation containing a recombinant monoclonal antibody, which comprises a 45 mM to 94 mM amino acid component having a histidine component of less than 5 mM, and a pH of 5.5 to 7.0.

<6-2> The aqueous solution of <6-1>, wherein the amino acid component other than the histidine component is at least selected from the group consisting of arginine, arginine hydrochloride and methionine 1 species.

<6-3> The aqueous solution of <6-1> or <6-2>, wherein the histidine component is at least one selected from the group consisting of histidine and histamine hydrochloride.

<6-4> An aqueous solution according to any one of <6-1> to <6-3> which contains an amino acid component of 75 mM to 93 mM.

<6-5> The aqueous solution of any one of <6-1> to <6-4> which contains 90 mM amino acid component.

<6-6> An aqueous solution according to any one of <6-1> to <6-5, wherein the pH is 5.5 to 6.7.

<6-7> The aqueous solution according to any one of <6-1> to <6-6>, which is used to dilute the concentration of the recombinant monoclonal antibody to 10 mg/mL to 200 mg/mL.

<6-8> The aqueous solution of any one of <6-1> to <6-7> which further contains a polyhydric alcohol.

<6-9> The aqueous solution of any one of <6-1> to <6-8> which further contains a surfactant.

<6-10> The aqueous solution of <6-9>, wherein the aforementioned surfactant is a nonionic surfactant.

<6-11> The aqueous solution of <6-10>, wherein the aforementioned nonionic surfactant is polysorbate or polyoxyethylene polyoxypropylene diol.

<6-12> The aqueous solution of <6-11>, wherein the polysorbate is at least one selected from the group consisting of polysorbate 80, polysorbate 40, and polysorbate 20.

<6-13> The aqueous solution of <6-11>, wherein the polyoxyethylene polyoxypropylene diol is polyoxyethylene (160) polyoxypropylene (30) diol.

<6-14> The aqueous solution of any one of <6-1> to <6-13>, wherein the immunoglobulin type of the recombinant monoclonal antibody is IgG1 derived from human.

<6-15> An aqueous solution of <6-14>, wherein the aforementioned IgG1 derived from human is Tocilizumab.

According to the present invention, it is possible to provide a recombinant recombinant monoclonal antibody liquid preparation which achieves at least one of dimer formation inhibition, decomposition product formation inhibition, biological activity reduction inhibition, and deamination inhibition during long-term storage. Or a recombinant recombinant monoclonal antibody liquid preparation which achieves at least one of dimer formation inhibition, decomposition product formation inhibition, bioactivity reduction inhibition, and deamination inhibition during long-term storage, and can be provided at low cost. preparation. Or a recombinant recombinant monoclonal antibody liquid preparation which achieves at least one of dimer formation inhibition, decomposition product formation inhibition, bioactivity reduction inhibition, and deamination inhibition during long-term storage, and can be provided at low cost. A liquid preparation containing a recombinant monoclonal antibody capable of exhibiting the kinetic viscosity of subcutaneous injection. Or a recombinant recombinant monoclonal antibody liquid preparation which achieves at least one of dimer formation inhibition, decomposition product formation inhibition, bioactivity reduction inhibition, and deamination inhibition during long-term storage, and can provide a practical vein A preparation for injection or a preparation for subcutaneous injection. Alternatively, a method for producing a recombinant recombinant monoclonal antibody liquid preparation which achieves at least one of dimer formation inhibition, decomposition product formation inhibition, bioactivity reduction inhibition, and deamination inhibition in long-term storage can be provided. Alternatively, it is possible to provide a method for stabilizing a recombinant monoclonal antibody liquid preparation which achieves both dimer formation inhibition, decomposition product formation inhibition, biological activity reduction inhibition, and deamination inhibition in long-term storage. Alternatively, an aqueous solution for formulating a formulation containing the recombinant monoclonal antibody can be provided.

Hereinafter, the present invention will be described in detail.

The liquid preparation containing the recombinant monoclonal antibody (hereinafter also referred to as "liquid preparation") in the present invention contains a recombinant monoclonal antibody of 10 mg/mL to 200 mg/mL, and a 45 mM to 94 mM having a histidine component of less than 5 mM. The amino acid component (total) and the pH is 5.5 to 7.0 or 5.5 to 6.7. Further, the liquid preparation may contain other ingredients. Also, liquid preparations can be provided at low cost. Further, the liquid preparation may be a preparation of any administration route.

The liquid preparation of the present invention contains a 45 mM to 94 mM amino acid component having a histidine component of less than 5 mM, and has a pH of 5.5 to 6.7, even if it contains a recombinant monoclonal antibody of 150 mg/mL to 200 mg/mL. In the liquid preparation, at least one of inhibition of dimer formation, inhibition of production of decomposition products, inhibition of reduction of biological activity, and inhibition of deamimination of the drug during long-term storage can be achieved, and the effect of exhibiting the kinetic viscosity of subcutaneous injection can be exhibited. Therefore, it is used as a preparation for subcutaneous injection. Further, since the liquid preparation of the present invention contains a low amount of an amino acid component of an amino acid component of 45 mM to 94 mM, it can be low in cost compared with the conventional liquid preparation containing the recombinant monoclonal antibody. provide.

Further, among the liquid preparations of the present invention, a liquid preparation containing a recombinant monoclonal antibody of 10 mg/mL to 30 mg/mL can be used as a dimer formation inhibition, a decomposition product formation inhibition, and a biological activity reduction in long-term storage. For example, at least one of inhibition and deamikation inhibition is a preparation for intravenous injection. A liquid preparation containing a recombinant monoclonal antibody of more than 30 mg/mL to less than 150 mg/mL is capable of inhibiting dimer formation, inhibiting production of decomposition products, and inhibiting reduction of biological activity during long-term storage during storage and storage. At least one of the inhibition of amiodalation. For example, it can be diluted and used at the time of use, and can be used as an intravenous preparation.

An aqueous solution for producing a liquid preparation containing a recombinant monoclonal antibody (hereinafter also referred to as "aqueous solution for producing an antibody preparation") of the present invention contains an amino acid component of 45 mM to 94 mM having a histidine component of less than 5 mM (total) And the pH is 5.5~7.0 or 5.5~6.7. Further, the aqueous solution for producing an antibody preparation may contain other components.

The aqueous solution for producing an antibody preparation of the present invention can achieve dimerization in long-term storage by containing a 45 mM to 94 mM amino acid component having a histidine component of less than 5 mM and having a pH of 5.5 to 7.0 or 5.5 to 6.7. At least one of inhibition of production of inhibitory substances, inhibition of reduction of biological activity, and inhibition of deamidation, when a preparation containing a recombinant monoclonal antibody is produced by using an aqueous solution for producing an antibody preparation, the recombinant monoclonal antibody can be stabilized. The effect. Further, any solution containing an antibody may be substituted with an aqueous solution for producing an antibody preparation by dialysis or the like to prepare a preparation containing a recombinant monoclonal antibody. Further, a preparation having a relatively high antibody concentration (for example, a preparation for subcutaneous injection) and an aqueous solution for producing an antibody preparation may be used to prepare a preparation having a relatively low concentration (for example, a preparation for intravenous injection). In this way, it is possible to prepare a common antibody preparation (for example, a subcutaneous injection preparation or an intravenous preparation) having a different concentration, an additive or the like, in a solution having the same composition, which is a common ingredient, or a manufacturing efficiency, and a cost. Reduction, etc., is industrially convenient.

In addition, in this specification, the numerical range represented by "~" is a range which shows the numerical value of the minimum value and the maximum value, respectively, including the value of the

Further, the unit of the molar concentration is M (molar), and the mM is 10 ^-3 mol/L.

In the present specification, the long-term storage may be, for example, stored at 2 ° C to 8 ° C for 1 year or longer, preferably at 2 ° C to 8 ° C for 2 years or more, and more preferably at 2 ° C to 8 ° C for 2 years. For 5 years, the best can be stored at 2 ° C ~ 8 ° C for 2 to 3 years. Alternatively, for example, it may be stored at 25 ° C for 6 months or more, and more preferably at 25 ° C for 1 year or more. Alternatively, for example, it may be stored at 40 ° C for 4 weeks or more, preferably at 40 ° C for 8 weeks or more, and more preferably at 40 ° C for 3 months to 6 months. Alternatively, for example, it can be stored at 50 ° C for 2 weeks. Alternatively, for example, it may be stored at 60 ° C for 1 to 2 weeks.

 (recombinant monoclonal antibody)  

Recombinant monoclonal antibody refers to an antibody produced by cells transformed with recombinant DNA technology. The recombinant monoclonal antibody is preferably as long as it is expressed or secreted in animal cells, but the type of the recombinant monoclonal antibody is not particularly limited. For example, a recombinant monoclonal antibody which can be used as a pharmaceutical is preferred.

Furthermore, the recombinant monoclonal antibody which can be contained in the liquid preparation of the present invention is not only a recombinant monoclonal antibody derived from an animal such as a human, a mouse or a rat, but also a recombinant individual such as a chimeric antibody or a humanized antibody. antibody. Further, the immunoglobulin type of the antibody is not particularly limited, and may be any of IgG, IgA, IgD, IgE, IgM, and the like such as IgG1, IgG2, IgG3, or IgG4. Among them, IgG1 derived from humans (for example, humanized IgG1 and fully human IgG1) is preferred. As IgG1 from humans, it is especially good for Tocilizumab.

Further, the recombinant monoclonal antibody also includes an antibody fragment such as Fv, Fab or F(ab) ₂ or a monovalent or bivalent or higher single chain in which a variable region of the antibody is linked to a linker such as a peptide linker. A low molecular weight antibody such as Fv (such as scFv, sc (Fv) ₂ or scFv dimer) or the like.

Such recombinant monoclonal antibodies can be formulated according to the methods described in International Publication No. 92/019759 and International Publication No. 2005/090405.

The type of the recombinant monoclonal antibody is not limited, and examples thereof include an anti-tumor necrosis factor (TNF) antibody or an anti-interleukin (IL) receptor antibody. Alternatively, for example, an anti-TNFα antibody, an anti-IL-6 receptor antibody, an anti-IL-17 receptor antibody, an anti-IL-5 antibody, an anti-IL-17 antibody, an anti-IL-17A antibody, an anti-IL-1β antibody, and an anti-IL-17 antibody may be mentioned. IL12/IL23-p40 antibody, anti-CD3 antibody, anti-CD20 antibody, anti-CD25 antibody, anti-CD30 antibody, anti-CD33 antibody, anti-CD52 antibody, anti-RANKL antibody, anti-SLAMF7 antibody, anti-CTLA-4 antibody, anti-VEGFR-2 antibody , anti-CCR4 antibody, anti-PD-1 antibody, anti-RS virus antibody, anti-α4 integrin antibody, anti-VEGF antibody, anti-EGFR antibody, anti-HER2 antibody, anti-PCSK9 antibody, anti-Dabigatran antibody, anti-IgE antibody, and anti-complement C5 antibody.

The type of the recombinant monoclonal antibody is not limited, and examples thereof include rheumatoid arthritis, juvenile idiopathic arthritis, Karber's disease, ankylosing spondylitis, Crohn's disease, ulcerative colitis, Pancreatitis, vasculitis, Kawasaki disease, systemic lupus erythematosus, dryness, dryness arthritis, Sjogren's disease, Steller's disease, multiple sclerosis, osteoporosis, bone disease, thrombosis, cancer For example, breast cancer, leukemia, ovarian cancer, melanoma, prostate cancer, pancreatic cancer, lymphoma, lung cancer, gastric cancer, renal cell carcinoma, colorectal cancer, mesothelioma, soft tissue sarcoma, multiple myeloma, etc.), cachexia , chronic rejection of transplanted organs and cells, heart failure, ischemic-induced severe arrhythmia, hypercholesterolemia, viral infections (eg RS virus infection, HIV infection, EBV infection, etc.), plasma cell hyperplasia, high immunoglobulin Proteinemia, anemia, renal cell proliferative nephritis, and asthma.

Examples of the recombinant monoclonal antibody include, but are not limited to, Trastuzumab, Rituximab, Palivizumab, Infliximab, Basiliximap, Gemtuzumab ozogamicin, Bevacizumab, Ibritumomab tiuxetan, Tocilizumab, Adalimumab, Cetuximab, Ranibizumab, Omalizumab, Eculizumab, Panitumumab, Ustekinumab, Golimumab, Canakinumab, Denosumab, Mogamulizumab, Ofatumumab, Pertuzumab, Trastuzumab emtansine, Brentuximab vedotin, Natalizumab, Nivolumab, Alemtuzumab, Secukinumab, Ramucirumab and Ipilimumab.

Further, recombinant monoclonal antibodies, preferably Tocilizumab, Trastuzumab, Rituximab, Palivizumab, Infliximab, Basiliximab, Bevacizumab, Adalimumab, Cetuximab, Ranibizumab, Omalizumab, Eculizumab, Panitumumab, Ustekinumab, Golimumab, Canakinumab, Denosumab, Mogamulizumab, Ofatumumab, Pertuzumab, Natalizumab, Nivolumab, Alemtuzumab, Secukinumab, Ramucirumab or Ipilimumab; more preferably Trastuzumab, Tocilizumab, Infliximab, Bevacizumab, Adalimumab, Ustekinumab, Golimumab or Denosumab; still more preferably Tocilizumab, Infliximab, Bevacizumab, Adalimumab or Denosumab.

Further, the recombinant monoclonal antibody is preferably Tocilizumab.

Tocilizumab, as long as it has the same amino acid sequence as the amino acid sequence of the H chain of the Actrama (registered trademark) (Gln1-Gly448) and the amino acid sequence of the L chain (Asp1-Cys214) . Further, the amino acid sequence of the H chain of Actemra (Glu1-Gly448) and the amino acid sequence of the L chain (Asp1-Cys214) are described in the Sequence Listing attached to International Publication No. 2005/090405.

Further, the N-terminal residue of the H chain may be pyroglutamic acid (pGlu) instead of glutamic acid. The C-terminal residue of the H chain may be up to 447 of proline (Pro), or may be a 449 amino acid residue of lysine (Gly) added to the amino acid (Lys) of No. 448. To replace 448 amino acid residues.

Antibody pharmaceuticals are generally higher in concentration than other biopharmaceuticals.

The liquid preparation of the present invention contains a recombinant monoclonal antibody of 10 mg/mL to 200 mg/mL. Moreover, from the viewpoint of sufficiently exerting the effects of the present invention, it is preferred to contain a recombinant monoclonal antibody of 170 mg/mL to 190 mg/mL, and more preferably a recombinant monoclonal antibody containing 180 mg/mL. Alternatively, it is preferably a recombinant monoclonal antibody containing 90 mg/mL to 110 mg/mL, more preferably a recombinant monoclonal antibody containing 100 mg/mL. Alternatively, it is preferably a recombinant monoclonal antibody containing 10 mg/mL to 30 mg/mL, more preferably a recombinant monoclonal antibody containing 15 mg/mL to 25 mg/mL, and still more preferably a recombinant monoclonal antibody containing 20 mg/mL. The recombinant monoclonal antibody used in the present invention is preferably not subjected to freeze-drying or reconstitution in the production method.

 (amino acid component)  

The liquid preparation of the present invention contains an amino acid component of 45 mM to 94 mM having a histidine component of less than 5 mM.

The type of the amino acid component other than the histidine component is not limited, and examples thereof include arginine, arginine hydrochloride, methionine, glycine, phenylalanine, aspartic acid, and glutamine. Acid, lysine, aspartame, tryptophan, cysteine and cysteine hydrochloride. In addition, from the viewpoint of at least one of inhibition of dimer formation, inhibition of production of decomposition products, suppression of reduction in biological activity, and inhibition of deamination, in the long-term storage, an amino acid component other than the histidine component Preferably, it is at least one selected from the group consisting of arginine, arginine hydrochloride, and methionine. Further, as the amino acid component other than the histidine component, it is preferred to contain arginine, arginine hydrochloride and methionine; more preferably, it contains arginine hydrochloride and methylamine. acid.

The histidine acid component is preferably at least one selected from the group consisting of histidine and histamine hydrochloride, more preferably histamine and histamine hydrochloride.

The liquid preparation of the present invention may contain a histamic acid of less than 5 mM in a liquid preparation, and preferably contains 1 mM or more and 4 mM or less of histidine, from the viewpoint that the pH can be adjusted to a preferred range. The component preferably contains 2 mM or more of a histidine component of 4 mM or less.

The amino acid component preferably contains a 45 mM to 94 mM amino acid component having a histidine component of less than 5 mM with respect to the entire liquid preparation. Further, it preferably contains an amino acid component of 75 mM to 93 mM, more preferably an amino acid component of 85 mM to 92 mM, an amino acid component particularly preferably containing 88 mM to 91 mM, and an amino acid component preferably containing 90 mM.

 (Polyol)  

The liquid preparation may further contain a polyol.

Examples of the polyhydric alcohol include propylene glycol, glycerin (glycerin), isoerythrocyanose, threitol, erythritol, erythritol, ribose, arabinose, arabitol, lyxose, maltitol, sorbitol, and sorbus. Sugar, glucose, mannose, mannitol, levulose, dextrose, maltose, trehalose, fructose, xylitol, inositol, galactose, xylose, fructose, sucrose, 1,2,6-hexanetriol . Among them, as the polyol, sucrose, trehalose and the like are preferable, and sucrose is preferred.

The liquid preparation may contain one or more of these polyols in combination.

Further, when the content of the amino acid component is less than 70 mM, it is preferred to use a combination of polyols from the viewpoint of suppressing the formation of a dimer.

The content of the polyol is not particularly limited, and may be appropriately determined from the viewpoint of the isomerization of the liquid preparation. For example, it may be 30 mg/mL to 80 mg/mL, 40 mg/mL to 70 mg/mL, and 50 mg/mL to 60 mg/mL.

 (surfactant)  

The liquid preparation may further contain a surfactant.

The surfactant may be selected from a cationic surfactant, an anionic surfactant, an amphoteric surfactant, a nonionic surfactant, etc., preferably a nonionic surfactant.

Examples of the surfactant include polyoxyethylene hardened castor oil (polyoxyethylene hardened castor oil 50, polyoxyethylene hardened castor oil 60, etc.), polyoxyethylene castor oil, castor oil fatty acid ethyl ester, nicotinic acid decylamine, and poly Oxyethylene sorbitan fatty acid ester (also known as polysorbate, Tween, etc., for example, polyoxyethylene (20) sorbitan (NIKKOL TL-10, polysorbate 20, Tween 20), single palm Acid polyoxyethylene (20) sorbitan (NIKKOL TP-10V, polysorbate 40, Tween 40), polyoxyethylene monostearate (20) sorbitan (NIKKOL TS-10MV, polysorbate 60, Tween 60), tristearate polyoxyethylene (20) sorbitan (NIKKOL TS-30V, polysorbate 65), monoisostearic acid polyoxyethylene (20) sorbitan (NIKKOL TI-10V), Monooleic acid polyoxyethylene (20) sorbitan (NIKKOL TO-10MV, polysorbate 80, Tween 80), trioleic acid polyoxyethylene (20) sorbitol (NIKKOL TO-30V, polysorbate 85 )), and polyoxyethylene polyoxypropylene diol (also known as Pluronic, Poloxamer, etc., such as polyoxyethylene (160) polyoxypropylene (30) diol (Pluronic F-68)). Among them, as the surfactant, especially polysorbate and polyoxyethylene polyoxypropylene diol are preferred; particularly preferred are polysorbate 80, polysorbate 40, polysorbate 20 and polyoxyethylene ( 160) Polyoxypropylene (30) diol; most preferably polysorbate 80.

One or two or more kinds of these surfactants may be combined in a liquid preparation.

In addition, when the content of the amino acid component is less than 70 mM, it is preferably a combination interface from the viewpoint of at least one of suppressing dimer formation, inhibiting decomposition product formation, reducing biological activity, and suppressing deamination. The active agent is used. Alternatively, it is preferred to add a surfactant as a stabilizer.

The content of the surfactant is not particularly limited and may be appropriately determined. For example, it may be 0.05 mg/mL to 20 mg/mL, 0.1 mg/mL to 10 mg/mL, 0.1 mg/mL to 5 mg/mL, 0.1 mg/mL to 0.5 mg/mL, and 0.15 mg/mL to 0.25 mg/mL.

The liquid formulation can be selected from any combination of an amino acid and a surfactant. For example, a combination of arginine and/or arginine hydrochloride, methionine, polysorbate 80; arginine and/or arginine hydrochloride, methionine, polysorbate may be selected. a combination of alcohol esters 60; a combination of arginine and/or arginine hydrochloride, methionine, polysorbate 40; arginine and/or arginine hydrochloride, methionine, Combination of polyoxyethylene (160) polyoxypropylene (30) diol; arginine and / or arginine hydrochloride, methionine, histidine and / or histidine hydrochloride, poly sorbate a combination of alcohol esters 80; a combination of arginine and/or arginine hydrochloride, methionine, histidine and/or histamine hydrochloride, polysorbate 60; arginine and/or Or a combination of arginine hydrochloride, methionine, histidine and / or histidine hydrochloride, polysorbate 40; arginine and / or arginine hydrochloride, methyl thiamine A combination of an acid, a histidine acid and/or a histidine hydrochloride, a polyoxyethylene (160) polyoxypropylene (30) diol.

 (other ingredients)  

The liquid preparation may contain, in addition to the recombinant monoclonal antibody and the amino acid component, other components necessary for the formulation of the liquid preparation. The other component may, for example, contain a dissolution aid, an isotonic agent, a preservative, an anti-adsorbent, a sulfur-containing reducing agent, and an antioxidant; preferably a dissolution aid.

The dissolution aid may, for example, be a surfactant, particularly a nonionic surfactant, and specifically, polyoxyethylene hardened castor oil (polyoxyethylene hardened castor oil 50, polyoxyethylene hardened castor oil 60, etc.), poly Sorbitol ester (polysorbate 80, polysorbate 40, polysorbate 20, etc.), polyoxyethylene sorbitan monolaurate, polyoxyethylene castor oil, castor oil fatty acid ethyl ester, and nicotine Acid amide and the like. When the same compound can be used as the surfactant and the dissolution aid, it is not necessary to use a separate dissolution aid, and it is preferable because the addition component other than the recombinant monoclonal antibody of the active ingredient can be minimized.

The isotonic agent may, for example, be a polyhydric alcohol, and examples thereof include salts such as sodium chloride, potassium chloride, and calcium chloride.

Examples of the preservative include methyl p-hydroxybenzoate, ethyl p-hydroxybenzoate, propyl p-hydroxybenzoate, sorbic acid, phenol, cresol, m-cresol, and chlorocresol.

Examples of the anti-adsorbing agent include human serum albumin, lecithin, dextran, hydroxypropyl vesin, methyl vesin, and polyethylene glycol.

Examples of the sulfur-containing reducing agent include N-ethinyl cysteine, N-ethylmercaptocysteine, lipoic acid, thiodiethylene glycol, thioethanolamine, thioglycerol, thiosorbitol, Thioglycolic acid and its salts, sodium thiosulfate, glutathione.

Examples of the antioxidant include erythorbic acid, dibutylhydroxytoluene, butylhydroxyanisole, α -tocopherol, tocopherol acetate, L-ascorbic acid and salts thereof, L-ascorbyl palmitate, and L-ascorbic acid stearic acid. Ester, sodium hydrogen sulfite, sodium sulfite, triamyl gallate, propyl gallate, disodium edetate (EDTA. 2Na), sodium pyrophosphate, sodium metaphosphate.

Furthermore, when preparing a liquid preparation containing a recombinant monoclonal antibody containing a recombinant monoclonal antibody of 30 mg/mL or less, for example, 20 mg/mL, 10 mg/mL, and 5 mg/mL, a histidine-containing component of less than 5 mM may be formulated. A liquid preparation containing a recombinant monoclonal antibody having a molecular weight of 45 mM to 94 mM and a pH of 5.5 to 7.0.

 (pH)  

The pH of the liquid preparation is preferably 5.5 to 7.0, more preferably 5.5 to 6.7, still more preferably 5.8 to 6.7, still more preferably 6.0 to 6.5, and particularly preferably 6.0 to 6.0 from the viewpoint of long-term stabilization effect. 6.2. Further, from the viewpoint of adjusting the dynamic viscosity of the liquid preparation to the range of subcutaneous injection or the stability of the secured preparation, the pH is preferably 5.5 to 6.7, more preferably 5.8 to 6.7, and even more preferably 6.0~6.5, especially good 6.0~6.2. In the case of the preparation for intravenous injection, that is, in the case of a concentration of 10 to 50 mg/mL, preferably in the case of a concentration of 10 to 30 mg/mL, the liquid preparation is from the viewpoint of the stability of the secured preparation. The pH is preferably 5.5 to 7.0, more preferably 5.5 to 6.7, still more preferably 6.0 to 6.5, and particularly preferably 6.0 to 6.2.

The measurement of pH can be carried out, for example, by a pH meter (model: HM-30G, manufactured by East Asia DKK Co., Ltd.). Further, the pH can be measured by the method described in the Japanese Pharmacopoeia of the 16th revision, for example, at room temperature (15 ° C to 25 ° C).

The pH of the liquid preparation can be adjusted by, for example, a histidine component contained in the liquid preparation, and other buffers can be used as needed to adjust the pH of the liquid preparation. Other buffering agents may, for example, be phosphate (sodium or potassium), sodium hydrogencarbonate, citrate (sodium or potassium), sodium acetate, sodium succinate, phosphoric acid, carbonic acid, citric acid, succinic acid, malic acid, gluconic acid, Glycine.

The method for producing a liquid preparation according to the present invention is a method comprising the following steps A to C, and optionally containing at least one of the steps D to F.

Step A: Step of preparing an antibody drug substance comprising any of the following steps A-1 to A-3

Step A-1: Step of melting the frozen antibody drug substance into a liquid state

Step A-2: Steps of preparing a liquid antibody drug substance containing any solvent

Step A-3: Steps of preparing the antibody raw material of the powder

Step B: adding an amino acid to the solvent to prepare an additive solution

Step C: mixing the raw materials prepared in the step A and the additive solution prepared in the step B

Step D: Step of making the pH of the solution adjusted in step C into 5.5 to 7.0 or 5.5 to 6.7

Step E: Step of filtering and sterilizing the prepared solution

Step F: Step of filling the solution prepared in step E and capping

The antibody drug substance is an antibody drug substance obtained by, for example, producing a recombinant monoclonal antibody and purifying it in a cell transformed with a recombinant DNA technique. An antibody bulk drug that has been preserved (e.g., refrigerated) can be used as long as it does not impair the activity of the antibody. Alternatively, an antibody drug substance obtained by producing a recombinant monoclonal antibody in a cell transformed with a recombinant DNA technique and purifying it. Or a commercially available frozen antibody bulk drug.

Step A is a step of preparing an antibody comprising any one of steps A-1 to A-3. Step A-1 is a step of melting the cryopreserved antibody drug substance into a liquid state. The frozen antibody drug substance can be melted into a liquid state by any method that does not impair the activity of the antibody. For example, it melts at room temperature, melts in a refrigerator, etc. The antibody drug substance may be frozen as long as it is a solvent. Although it is preferably an aqueous solution for producing a liquid preparation containing a recombinant monoclonal antibody of the present invention, it may be, for example, water, physiological saline, glucose solution, any buffer solution, ethanol solution, or other pharmaceutically usable solvent. The melted antibody bulk drug can also be subjected to solvent exchange using any solvent dialysis. Step A-2 is a step of preparing a liquid antibody drug substance containing any solvent. The liquid antibody drug substance can also be subjected to solvent exchange using any solvent dialysis. Step A-3 is a step of preparing an antibody raw material for powder.

Step B is a step of preparing an additive solution containing an amino acid. For example, an amino acid may be added to a solvent (for example, water, physiological saline, glucose solution, any buffer solution, ethanol solution, or other pharmaceutically acceptable solvent) as an additive solution.

Step C is a step of mixing the raw material drug melted in the above step A with the additive solution prepared in the above step B. The final concentration of the amino acid component in the entire liquid is adjusted, for example, to 45 mM to 94 mM having a histidine acid component of less than 5 mM.

The method for producing a liquid preparation of the present invention may further comprise the following step D, step E, and step F.

Step D is a step of adjusting the pH of the solution prepared in the above step C to 5.5 to 7.0. Alternatively, step D is a step of adjusting the pH of the solution prepared in the above step C to 5.5 to 6.7. The pH of the liquid preparation can also be adjusted to pH 5.5 to 7.0 or 5.5 to 6.7 by the step C, for example, by adjusting the histidine component contained in the liquid preparation, but other additives can also be used to adjust the liquid preparation. pH.

Step E is a step of sterilizing the solution prepared in the step C or the step D by filtration. For filter sterilization, for example, a sputum filter made of polyvinylidene fluoride can be used. Alternatively, other sterilization methods that are generally used may be used.

Step F is a step of filling the solution prepared in the step C, the step D or the step E, and capping. Alternatively, it can be filled in an ampoule or pre-filled syringe or as a premixed bag.

In the liquid preparation of the present invention, an antibody raw material (for example, a frozen antibody raw material drug) may be prepared, and a solvent (for example, water, physiological saline, glucose solution, any buffer solution, ethanol solution, or the like) may be added thereto. After the pharmaceutically acceptable solvent), an amino acid is added to produce.

The component optionally contained in the liquid preparation of the present invention may be added to the additive solution in advance, or may be added after the antibody is added to the solvent, together with the amino acid or before the addition of the amino acid.

The liquid preparation of the present invention can be administered, for example, by injection (subcutaneous injection, intravenous injection, intramuscular injection, etc.), transdermally, transmucosally, nasally, or pulmonaryly.

When subcutaneous injection is performed, the amount of recombinant antibody administered per time is a large amount of 150 mg/mL to 200 mg/mL, and the amount of the injection is limited. The liquid preparation of the present invention is a recombinant monoclonal antibody containing 150 mg/mL to 200 mg/mL and exhibits an kinetic viscosity capable of subcutaneous injection. Therefore, in the present invention, a liquid preparation containing a recombinant monoclonal antibody of 150 mg/mL to 200 mg/mL is preferably used as a subcutaneous injection from the viewpoint of exerting the effects of the present invention. Subcutaneous injection is not only carried out by experts who are medical practitioners, but may be self-injected by the patient. There are many patients who are uneasy about self-injection, preferably dynamic viscosity. Low, and its stability over time is excellent. Further, in the present invention, a liquid preparation containing a recombinant monoclonal antibody of 10 mg/mL to 30 mg/mL is preferably used as a preparation for intravenous injection. Moreover, when these preparations are manufactured, the preparation for subcutaneous injection and the preparation for intravenous injection having different concentrations can be prepared with a solution having the same composition, which is industrially advantageous.

The dynamic viscosity of the liquid preparation of the present invention (for example, a preparation having an antibody concentration of 150 mg/mL to 200 mg/mL) is preferably, for example, 6 mm ² /s to 15 mm ² /s, more preferably after preparation of the liquid preparation. 7mm ² /s~14mm ² /s, and more preferably 8mm ² /s~12mm ² /s.

The dynamic viscosity can also be measured by a Ubellohde type viscometer (the 16th revision of the Japanese Pharmacopoeia general test method 2.53 viscosity measurement method 1). Alternatively, it is also possible to measure the tension generated in the plunger when the injection needle is attached to the glass syringe, and to apply the tension to the same measurement of the dynamic viscosity of 2 mm 2 /s to 20 mm 2 . The viscosity of the /s viscometer is corrected by the calibration curve obtained by the tension and dynamic viscosity obtained by the standard solution (Grease, Japan).

The dimer product and the low molecular weight decomposed product can be measured, for example, by a high-speed liquid chromatography system (Prominence, manufactured by Shimadzu Corporation) by size screening chromatography.

Biological activity can be assessed by measuring binding to antigen. For example, in Tocilizumab, the IL-6 binding inhibitory effect on soluble IL-6 receptor by Tocilizumab can be determined by ELISA. Alternatively, the biological activity assay assesses the effect of the antigen or the inhibition or promotion of the in vivo response to the antigenic vector. For example, in Tocilizumab, the effect of Tocilizumab on the expression of IL-6 activity of the membrane-bound IL-6 receptor vector can be assessed by the proliferation inhibition of IL-6-dependent cells.

The amidoxime can be measured, for example, by ion exchange chromatography using a high-speed liquid chromatography system (Prominence, manufactured by Shimadzu Corporation).

The invention is further illustrated by the following examples, which are not to be construed as limited. In the present specification, the expression "%" means % by mass unless otherwise specified.

 [Examples]    (Example 1)    Confirmation of the stability of liquid preparations  

For the liquid preparation containing the recombinant monoclonal antibody (180 mg/mL in Tocilizumab), the effect of the formulation of the present invention containing the 94 mM amino acid component on the stabilization of each pH was evaluated.

In this discussion, in order to confirm the stabilization effect of the liquid preparation, the evaluation samples No. 1 to No. 5 were prepared. The basis of each evaluation sample is as follows. Further, "-" in the compositions in Tables 1 and 5 indicates that it was not blended. Further, the Tocilizumab used in the examples can be prepared according to the methods described in International Publication No. 92/019759, International Publication No. 2005/090405, International Publication No. 99/063058, and International Publication No. 2002/072615.

In order to evaluate the stability of the liquid preparation, each evaluation sample was filled in a 0.5 mL to 2 mL small glass bottle, and the thermal acceleration test of each evaluation sample was performed (60 ° C - 2 weeks, 50 ° C - 2 weeks, and 40 ° C - 4 weeks). save). Then, the purity of the recombinant monoclonal antibody before and after the thermal acceleration was confirmed by size exclusion chromatography (SEC) and ion exchange chromatography (IEX). Also, the usability of the liquid preparation was evaluated by the dynamic viscosity. The conditions for analysis of size screening chromatography (SEC), ion exchange chromatography (IEX), and dynamic viscosity are as follows.

 [Size screening chromatography]  

The evaluation sample was diluted with the mobile phase to have a protein concentration of 1 mg/mL as an evaluation sample solution.

20 μL of the evaluation sample solution was tested by liquid chromatography under the following conditions, and the peak area of the polymer division, the main division, and the low molecular division was measured by an automatic analysis method, and the amount (%) was determined.

 Analysis condition  

Column: TSKgel G3000SWxl 7.8mm I.D.×30cm (made by Tosoh Corporation)

Protection column: TSKgel guard column SW _XL 6.0mm ID × 4cm (made by Tosoh Corporation)

Mobile phase: phosphate buffer pH 6.8 (20 mmol/L phosphate buffer containing pH 6.8 of 300 mmol/L sodium chloride)

Evaluation of sample injection amount: about 20 μg based on recombinant monoclonal antibody

Flow rate: 0.5mL/min

Detection wavelength: 280nm

 Computational  

Total area of each peak = peak area of main division + peak area of high polymer division + peak area of low molecular division

Polymer classification (%) = (total of each peak area of the polymer division / total area of each peak) × 100

Low molecular discrimination (%) = (total of each peak area of low molecular division / total area of each peak) × 100

 [ion exchange chromatography]  

The evaluation sample was diluted with mobile phase A to have a protein concentration of 2 mg/mL as an evaluation sample solution.

For evaluation of 20 μL of the sample solution, the liquid chromatography method was used to carry out the test under the following conditions, and the peak area of the acid separation, the main division, and the alkaline division was measured by an automatic analysis method, and the amount (%) was determined.

 Analysis condition  

Column: WCX-10 4mm I.D.×25cm (Thermo Fisher Scientific)

Protection column: WCX-10G 4mm I.D.×5cm (Thermo Fisher Scientific)

Mobile phase A: phosphate buffer pH 7.0 (10 mmol/L sodium phosphate buffer at pH 7.0)

Mobile phase B: pH 7.0 phosphate buffer (10 mmol/L sodium phosphate buffer containing 500 mmol/L sodium chloride pH 7.0)

Evaluation of sample injection amount: about 40 μg based on recombinant monoclonal antibody

Flow rate: 1.0mL/min

Detection wavelength: 280nm

 Computational  

Total area of each peak = peak area of main distinction + area of each peak of acid distinction + peak area of each alkaline division

Acidity (%) = (the total of the peak areas of each acid distinction / the total area of each peak) × 100

Alkaline discrimination (%) = (total of peak areas of each alkaline division / total area of each peak) × 100

The evaluation sample was directly used as the evaluation sample solution.

The tension generated in the plunger when the evaluation sample solution was sucked in a state where the injection needle (22 G × 1) was attached to a glass syringe (0.25 mL) was measured. And the obtained tension is fitted to a calibration curve made by the tension and dynamic viscosity obtained by measuring the viscous meter calibration standard solution (Grease, Japan) having the dynamic viscosity of 2 mm ² /s to 20 mm ² /s. Get the viscosity. The measured temperature of the dynamic viscosity was 20 °C.

The evaluation results of the size screening chromatography obtained in this example are shown in Table 2. The evaluation results of the ion exchange chromatography are shown in Table 3. The evaluation results of the dynamic viscosity are shown in Table 4.

The formulation containing the amino acid component of 94 mM and adjusting the pH to 6 (evaluation sample No. 5) has a small amount of amino acid and a content of histidine component, but at 60 ° C for 2 weeks. In the thermal acceleration test after storage at 50 ° C for 2 weeks and 40 ° C for 4 weeks, the amount of the polymer containing the dimer or the like and the low molecular weight of the decomposition product and the like are compared with the total amount of the amino acid and the histamine. The content of the acid component was the same as that of a large amount of the evaluation sample No. 1.

The product of the deamidamine reaction which adversely affects the stability of the protein preparation, i.e., the deaminating amine, is detected by acid separation in ion exchange chromatography. When the sample No. 5 was evaluated, it was confirmed that the increase in the acidity of the deaminated amine contained in the ion exchange chromatography was suppressed similarly to the evaluation sample No. 1. Further, regarding the alkaline separation in the ion exchange chromatography, the evaluation sample No. 5 also confirmed that the increase was suppressed to the same extent as the evaluation sample No. 1. This is considered to be that the decomposition product is reduced and the stability is improved. In addition, in the case where the pH was adjusted to 5 (evaluation sample No. 2) and the pH was adjusted to 6.8 (evaluation sample Nos. 3 and 4), the evaluation was made in comparison with the polymer containing dimer. In sample No. 5, an increase tendency was observed, and an increase tendency was observed in the acid division containing a deaminating substance or the like. Therefore, it is apparent that the pH range is an important management item. In particular, for example, under severe conditions such as 60 ° C, for example, under acidic conditions such as pH 5.0, it is confirmed that the increase in the molecular division and the increase in the basicity are observed, and the tendency to deteriorate in stability is observed.

From the evaluation results of the presence or absence of the addition of histidine (evaluation samples Nos. 3 and 4), it is apparent that the histidine component is important from the viewpoint of adjusting the pH of the liquid preparation to the desired pH range. ingredient.

Further, the dynamic viscosity of the sample No. 5 was evaluated, and the dynamic viscosity of the sample No. 1 was lower than that of the evaluation sample No. 1, and the change was small, so that the usability at the time of subcutaneous injection was improved.

Thus, it is apparent that even if the content of the amino acid component is 94 mM or less, a liquid preparation exhibiting the kinetic viscosity of subcutaneous injection can be prepared by adjusting the pH to a range of 5.5 to 6.7.

 (Example 2)  

Confirmation of the stability effect when the content of the amino acid component is further reduced

The effect of stabilization was evaluated for a liquid preparation containing a recombinant monoclonal antibody (180 mg/mL in Tocilizumab) when the content of the amino acid component was lower than that of 94 mM.

In this discussion, in order to evaluate the possibility of further reducing the concentration of the amino acid component, evaluation samples No. 6-1 to No. 8 were prepared. The basis of each evaluation sample is as follows.

In order to evaluate the stability of the liquid preparation, each evaluation sample was filled in a 0.5 mL to 2 mL small glass bottle, and the thermal acceleration test of each evaluation sample was performed (60 ° C - 1 week, 50 ° C - 2 weeks, 40 ° C - 8 weeks). And 25 ° C -4 months to save). The purity of the recombinant monoclonal antibody before and after thermal acceleration was evaluated by size screening chromatography and ion exchange chromatography, and the usability was evaluated by dynamic viscosity. The analysis conditions are as shown in Example 1.

The evaluation results of the size screening chromatography obtained in the present example are shown in Table 6. The evaluation results of the ion exchange chromatography are shown in Table 7, and the evaluation results of the dynamic viscosity are shown in Table 8.

Regardless of the prescription, as shown in Table 6, when compared with the evaluation sample (evaluation sample No. 1) in which the concentration of the amino acid component shown in Example 1 is more than 94 mM, the size sieve In the evaluation of the chromatographic method, it was confirmed that the increase in the molecular division containing the dimer or the like can be suppressed by the evaluation of 50 ° C - 2 weeks from the test in which the thermal stability is the same condition. In addition, it was confirmed that the increase in the low molecular weight of the decomposition product or the like was suppressed in the same manner as the polymer separation.

As a result, it is apparent that the content of the amino acid component is 93 mM or less, and the formation of a dimer and the formation of a decomposition product in the liquid preparation can be sufficiently suppressed.

Further, as shown in Table 7, it was confirmed that the acidity of the deacetylated amine was evaluated in the evaluation of 50 ° C - 2 weeks from the test in which the thermal stability was the same condition as compared with the evaluation sample No. 1. The generation inhibition effect of the distinction is further improved. In the same manner, it was confirmed that the alkaline differentiation generation suppressing effect was also improved. Further, regarding the dynamic viscosity, no significant increase was observed in any of the evaluation samples, and it was confirmed that the subcutaneous injection showed no problem of usability.

When the concentration of the amino acid component was lower than 90 mM (evaluation samples No. 7 and 8), it was confirmed that the concentration of the amino acid component was lower than 90 mM, and in the accelerated condition, the size was observed. In the evaluation of the screening chromatography, the polymer containing the dimer has a tendency to increase, but by adding sucrose (purified white sugar), the effect of the increase can be suppressed.

Further, even when 55.66 mg/mL of sucrose was added, no significant increase in dynamic viscosity was observed, and it was confirmed that subcutaneous injection showed no problem of useability. It is apparent that the effect of suppressing the formation of a dimer can be exhibited by combining an amino acid component and sucrose.

 (Example 3)    Confirmation of the stability of liquid preparations  

For the liquid preparation containing the recombinant monoclonal antibody (20 mg/mL in Tocilizumab), the effect of the pH of each of the formulations of the present invention containing 94 mM of the amino acid component on the stabilization was evaluated.

In this discussion, in order to confirm the stabilization effect of the liquid preparation, an evaluation sample No. 9 to No. 15 was prepared. The basis of each evaluation sample is as follows. Further, "-" in the compositions in Tables 9 and 10 indicates that it was not blended. Further, the Tocilizumab used in the examples can be prepared according to the methods described in International Publication No. 92/019759, International Publication No. 2005/090405, International Publication No. 99/063058, and International Publication No. 2002/072615.

In order to evaluate the stability of the liquid preparation, each evaluation sample was filled in a 0.5 mL to 2 mL small glass bottle, and the thermal acceleration test of each evaluation sample was performed (60 ° C - 2 weeks, 50 ° C - 2 weeks, and 40 ° C - 4 weeks). save). Then, the purity of the recombinant monoclonal antibody before and after the thermal acceleration was confirmed by size exclusion chromatography (SEC) and ion exchange chromatography (IEX). Also, the usability of the liquid preparation was evaluated by the dynamic viscosity. The conditions for analysis of size screening chromatography (SEC), ion exchange chromatography (IEX), and dynamic viscosity are as follows. The same evaluation results as in the first embodiment can be expected.

 [Size screening chromatography]  

The evaluation sample was diluted with the mobile phase to have a protein concentration of 1 mg/mL as an evaluation sample solution.

20 μL of the evaluation sample solution was tested by liquid chromatography under the following conditions, and the peak area of the polymer division, the main division, and the low molecular division was measured by an automatic analysis method, and the amount (%) was determined.

 Analysis condition  

Column: TSKgel G3000SWxl 7.8mm I.D.×30cm (made by Tosoh Corporation)

Protection column: TSKgel guard column SW _XL 6.0mm ID × 4cm (made by Tosoh Corporation)

Mobile phase: phosphate buffer pH 6.8 (20 mmol/L phosphate buffer containing pH 6.8 of 300 mmol/L sodium chloride)

Evaluation of sample injection amount: about 20 μg based on recombinant monoclonal antibody

Flow rate: 0.5mL/min

Detection wavelength: 280nm

 Computational  

Total area of each peak = peak area of main division + peak area of high polymer division + peak area of low molecular division

Polymer classification (%) = (total of each peak area of the polymer division / total area of each peak) × 100

Low molecular discrimination (%) = (total of each peak area of low molecular division / total area of each peak) × 100

 [ion exchange chromatography]  

The evaluation sample was diluted with mobile phase A to have a protein concentration of 2 mg/mL as an evaluation sample solution.

20 μL of the evaluation sample solution was tested by liquid chromatography under the following conditions, and the peak area of the acid classification, the main division, and the alkaline division was measured by an automatic analysis method, and the amount (%) was determined.

 Analysis condition  

Column: WCX-10 4mm I.D.×25cm (Thermo Fisher Scientific)

Protection column: WCX-10G 4mm I.D.×5cm (Thermo Fisher Scientific)

Mobile phase A: phosphate buffer pH 7.0 (10 mmol/L sodium phosphate buffer at pH 7.0)

Mobile phase B: pH 7.0 phosphate buffer (10 mmol/L sodium phosphate buffer containing 500 mmol/L sodium chloride pH 7.0)

Evaluation of sample injection amount: about 40 μg based on recombinant monoclonal antibody

Flow rate: 1.0mL/min

Detection wavelength: 280nm

 Computational  

Total area of each peak = peak area of main distinction + area of each peak of acid distinction + peak area of each alkaline division

Acidity (%) = (the total of the peak areas of each acid distinction / the total area of each peak) × 100

Alkaline discrimination (%) = (total of peak areas of each alkaline division / total area of each peak) × 100

 [Dynamic Viscosity Measurement]  

The evaluation sample was directly used as the evaluation sample solution.

The tension generated in the plunger when the evaluation sample solution was sucked in a state where the injection needle (22 G × 1) was attached to a glass syringe (0.25 mL) was measured. And the obtained tension is fitted to a calibration curve made by the tension and dynamic viscosity obtained by measuring the viscous meter calibration standard solution (Grease, Japan) having the dynamic viscosity of 2 mm ² /s to 20 mm ² /s. Get the viscosity. The measured temperature of the dynamic viscosity was 20 °C.

 (Example 4)  

Confirmation of the stability effect when the content of the amino acid component is further reduced

The stability effect was evaluated for a liquid preparation containing a recombinant monoclonal antibody (20 mg/mL in Tocilizumab) when the content of the amino acid component was further lowered than 94 mM.

In this discussion, in order to evaluate the possibility of further reducing the concentration of the amino acid component, evaluation samples No. 16-1 to No. 18 were prepared. The basis of each evaluation sample is as follows. The same evaluation results as in Example 2 can be expected.

In order to evaluate the stability of the liquid preparation, each evaluation sample was filled in a 0.5 mL to 2 mL small glass bottle, and the thermal acceleration test of each evaluation sample was performed (60 ° C - 1 week, 50 ° C - 2 weeks, 40 ° C - 8 weeks). And 25 ° C -4 months to save). The purity of the recombinant monoclonal antibodies before and after thermal acceleration was evaluated by size screening chromatography and ion exchange chromatography, and the usability was evaluated by dynamic viscosity. The analysis conditions are as shown in Example 1.

From Examples 3 and 4, it can be considered that, for example, compared with pH 5.5 to 7.0, at pH 5.0 and pH 7.2, sufficient long-term stabilization effect of the preparation was not observed, and certain stability was observed at a specific pH. The effect is expected.

 [Industrial Applicability]  

The preparation of the present invention is a recombinant single antibody liquid preparation which is stable in the prevention of dimer formation and inhibition of deamidation during long-term storage, and is industrially useful because it can provide a preparation at low cost.

The preparation of the present invention is a recombinant monoclonal antibody liquid preparation which achieves dimer formation inhibition and deamidation inhibition during long-term storage, and is capable of providing a dynamic viscosity capable of subcutaneous injection at a low cost. The liquid preparation of recombinant monoclonal antibody is industrially useful.

Claims (20)

 A liquid preparation containing a recombinant monoclonal antibody comprising a recombinant monoclonal antibody of 10 mg/mL to 200 mg/mL, and an amino acid component of 45 mM to 94 mM having a histidine component of less than 5 mM, and having a pH of 5.5 to 7.0.    A liquid preparation containing a recombinant monoclonal antibody comprising a recombinant monoclonal antibody of 10 mg/mL to 200 mg/mL, and an amino acid component of 45 mM to 94 mM having a histidine component of less than 5 mM, and having a pH of 5.5 to 6.7.    The liquid preparation according to claim 1 or claim 2, wherein the amino acid component other than the histidine component is at least one selected from the group consisting of arginine, arginine hydrochloride and methionine .    The liquid preparation according to any one of claims 1 to 3, wherein the histidine component is at least one selected from the group consisting of histidine and histamine hydrochloride.    The liquid preparation according to any one of claims 1 to 4, which contains an amino acid component of 75 mM to 93 mM.    The liquid preparation according to any one of claims 1 to 5, which contains a 90 mM amino acid component.    The liquid preparation according to any one of claims 1 to 6, wherein the pH is from 5.8 to 6.7.    The liquid preparation according to any one of claims 1 to 7, wherein the pH is from 6.0 to 6.5.    The liquid preparation according to any one of claims 1 to 8, which contains a recombinant monoclonal antibody of 10 mg/mL to 30 mg/mL.    A liquid preparation according to claim 9, which comprises a recombinant antibody of 20 mg/mL.    The liquid preparation according to any one of claims 1 to 10, further comprising a polyol.    The liquid preparation according to any one of claims 1 to 11, further comprising a surfactant.    A liquid preparation according to claim 12, wherein the aforementioned surfactant is a nonionic surfactant.    The liquid preparation of claim 13, wherein the aforementioned nonionic surfactant is polysorbate or polyoxyethylene polyoxypropylene diol.    The liquid preparation according to claim 14, wherein the polysorbate is at least one selected from the group consisting of polysorbate 80, polysorbate 40, and polysorbate 20.    The liquid preparation of claim 14, wherein the polyoxyethylene polyoxypropylene diol is a polyoxyethylene (160) polyoxypropylene (30) diol.    The liquid preparation according to any one of claims 1 to 16, wherein the recombinant monoclonal antibody is a recombinant monoclonal antibody, a chimeric antibody, a humanized antibody selected from the group consisting of an animal (human, mouse, rat, etc.) At least one of the group consisting of an antibody fragment and a low molecular weight antibody.    The liquid preparation according to any one of claims 1 to 17, wherein the immunoglobulin type of the recombinant monoclonal antibody is selected from the group consisting of IgG (IgG1, IgG2, IgG3, IgG4), IgA, IgD, IgE, and IgM At least one of the groups.    The liquid preparation according to any one of claims 1 to 18, wherein the immunoglobulin type of the recombinant individual is IgG1 (humanized antibody and fully human antibody) derived from human.    The liquid preparation according to any one of claims 1 to 19, wherein the recombinant monoclonal antibody is selected from the group consisting of Tocilizumab, Trastuzumab, Rituximab, Palivizumab, Infliximab, Basiliximap, Gemtuzumab ozogamicin, Bevacizumab, Ibritumomab tiuxetan, Adalimumab, Cetuximab, Ranibizumab At least one of the group consisting of Omalizumab, Eculizumab, Panitumumab, Usteninumab, Golimumab, Canakinumab, Denosumab, Mogamulizumab, Ofatumumab, Pertuzumab, Trastuzumab emtansine, Brentuximab vedotin, Natalizumab, Nivolumab, Alemtuzumab, Secukinumab, Ramucirumab and Ipilimumab.