WO2017057644A1 - Liquid formulation containing high concentration of antibodies - Google Patents

Liquid formulation containing high concentration of antibodies Download PDF

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Publication number
WO2017057644A1
WO2017057644A1 PCT/JP2016/078942 JP2016078942W WO2017057644A1 WO 2017057644 A1 WO2017057644 A1 WO 2017057644A1 JP 2016078942 W JP2016078942 W JP 2016078942W WO 2017057644 A1 WO2017057644 A1 WO 2017057644A1
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Prior art keywords
liquid preparation
recombinant monoclonal
amino acid
monoclonal antibody
histidine
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PCT/JP2016/078942
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French (fr)
Japanese (ja)
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龍 奥脇
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持田製薬株式会社
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Publication of WO2017057644A1 publication Critical patent/WO2017057644A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/08—Solutions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
    • A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/20—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing sulfur, e.g. dimethyl sulfoxide [DMSO], docusate, sodium lauryl sulfate or aminosulfonic acids

Definitions

  • the present invention relates to a liquid preparation containing a high concentration antibody.
  • Recombinant monoclonal antibodies such as tocilizumab are currently used as active ingredients in pharmaceuticals.
  • the dose of the recombinant monoclonal antibody per administration may be large.
  • the amount of injection when using a preparation for subcutaneous injection capable of self-injection, the amount of injection once There are limitations. Therefore, it may be necessary to increase the concentration of the recombinant monoclonal antibody contained in the preparation.
  • Patent Document 1 a pharmaceutical formulation containing a specific amount of arginine, methionine, polysorbate
  • Patent Document 2 a pharmaceutical formulation containing a specific amount of arginine hydrochloride, histidine, polysorbate
  • the content of the recombinant monoclonal antibody is 150 mg / mL or more
  • the content of amino acid components such as arginine is 100 mM or more.
  • the present invention is a stable high-concentration recombinant monoclonal antibody liquid preparation that achieves dimer formation suppression and deamidation suppression during long-term storage, and also exhibits high-concentration recombination that exhibits kinematic viscosity that enables subcutaneous injection
  • An object is to provide a liquid preparation containing a monoclonal antibody at low cost.
  • the present invention is as follows. ⁇ 1> High-concentration recombination containing 150 mg / mL to 200 mg / mL recombinant monoclonal antibody and 45 mM to 94 mM amino acid component having a histidine component of less than 5 mM and a pH of 5.5 to 6.7 Monoclonal antibody-containing liquid preparation. ⁇ 2> The liquid preparation according to ⁇ 1>, 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.
  • ⁇ 3> The liquid formulation according to ⁇ 1> or ⁇ 2>, wherein the histidine component is at least one selected from the group consisting of histidine and histidine hydrochloride.
  • ⁇ 4> The liquid preparation according to any one of ⁇ 1> to ⁇ 3>, comprising an amino acid component of 75 mM to 93 mM.
  • ⁇ 5> The liquid preparation according to any one of ⁇ 1> to ⁇ 4>, containing 90 mM amino acid component.
  • ⁇ 6> The liquid preparation according to any one of ⁇ 1> to ⁇ 5>, wherein the pH is 5.8 to 6.4.
  • ⁇ 7> The liquid preparation according to any one of ⁇ 1> to ⁇ 6>, wherein the pH is 6.0 to 6.2.
  • ⁇ 8> The liquid preparation according to any one of ⁇ 1> to ⁇ 7>, comprising 170 mg / mL to 190 mg / mL recombinant monoclonal antibody.
  • ⁇ 9> The liquid preparation according to any one of ⁇ 1> to ⁇ 8>, which contains 180 mg / mL recombinant monoclonal antibody.
  • ⁇ 10> The liquid preparation according to any one of ⁇ 1> to ⁇ 9>, further containing a polyol.
  • Recombinant monoclonal antibodies are tocilizumab, trastuzumab, rituximab, rituximab, rituximab, rituximab, rituximab, rituximab, rituximab, rituximab, rituximab, rituximab, rituximab, rituximab, rituximab, rituximab, rituximab, rituximab, rituximab, rituximab, rituximab At least one liquid selected from the group consisting of canakinumab, denosumab, mogamulizumab, ofatumumab, pertuzumab, trastuzumab emtansine, brentuximab vedotin, natalizumab, n
  • the present invention is a stable high-concentration recombinant monoclonal antibody liquid preparation that realizes dimer formation inhibition and deamidation inhibition during long-term storage, and has a high concentration that exhibits kinematic viscosity that enables subcutaneous injection.
  • a liquid preparation containing a recombinant monoclonal antibody can be provided at low cost.
  • a liquid preparation containing a high-concentration recombinant monoclonal antibody in the present invention (hereinafter also referred to as “liquid preparation”) comprises a recombinant monoclonal antibody of 150 mg / mL to 200 mg / mL and 45 mM to 94 mM having a histidine component of less than 5 mM.
  • the amino acid components (total) are contained, and the pH is 5.5 to 6.7.
  • the liquid preparation may contain other components.
  • the liquid preparation in the present invention contains an amino acid component of 45 mM to 94 mM in which the histidine component is less than 5 mM, and the pH is adjusted to the range of 5.5 to 6.7, and as high as 150 mg / mL to 200 mg / mL. Even if it contains a recombinant monoclonal antibody at a concentration, it has the effect of suppressing dimer formation and deamidation during long-term storage in liquid formulations, and exhibiting kinematic viscosity that enables subcutaneous injection Can be played.
  • the liquid preparation in the present invention contains a low-dose amino acid component of 45 mM to 94 mM amino acid component, it can be provided at a lower cost than conventional liquid preparations containing a high concentration recombinant monoclonal antibody.
  • a numerical range indicated by using “to” indicates a range including the numerical values described before and after “to” as the minimum value and the maximum value, respectively.
  • the unit of molar concentration is M (molar), and mM is 10 ⁇ 3 mol / L.
  • long-term storage includes, for example, storage at 2 ° C. to 8 ° C. for 1 year or longer, preferably storage at 2 ° C. to 8 ° C. for 2 years or longer, more preferably 2 It can be stored for 2 to 3 years at -8 to 8 ° C.
  • a recombinant monoclonal antibody is an antibody produced by cells transformed by applying recombinant DNA technology.
  • the recombinant monoclonal antibody is preferable if it is expressed or secreted in animal cells, but the type of the recombinant monoclonal antibody is not particularly limited.
  • a recombinant monoclonal antibody that can be used as a pharmaceutical is preferable.
  • Recombinant monoclonal antibodies that can be contained in the liquid preparation of the present invention include not only recombinant monoclonal antibodies derived from animals such as humans, mice, and rats, but also recombinant monoclonal antibodies such as chimeric antibodies and humanized antibodies.
  • the immunoglobulin class of the antibody is not particularly limited, and any class such as IgG such as IgG1, IgG2, IgG3, and IgG4, IgA, IgD, IgE, and IgM may be used.
  • Recombinant monoclonal antibodies include antibody fragments such as Fv, Fab, F (ab) 2, and monovalent or bivalent or more single-chain Fvs (variable regions of antibodies bound by a linker such as a peptide linker). scFv, sc (Fv) 2 , Diabodies such as scFv dimer) and the like are also included. These recombinant monoclonal antibodies can be prepared according to the methods described in International Publication No. 92/019759 and International Publication No. 2005/090405.
  • Examples of the recombinant monoclonal antibody include, but are not limited to, for example, trastuzumab, rituximab, palivizumab, infliximab, basiliximab, gemtuzumab ozogamicin, bevacizumab, ibritumomab tiuxetane, tocilizumab, adalimumab, , Ranibizumab, Omalizumab, Eculizumab, Panitumumab, Usutekinumab, Golimumab, Kanakinumab, Denosumab, Mogamulizumab, Offatumumab, Pertuzumab, Trastuzumab Emtansin, Brentuximab, Beduminumab
  • Recombinant monoclonal antibodies include tocilizumab, trastuzumab, rituximab, paclitumumab, infliximab, infliximab, basiliximab, bevacizumab, adalimumab, cetuximab, ranibizumab, omalizumab, eculizumab, panitumabum , Nivolumab, alemtuzumab, secukinumab, ramcilumab or ipilimumab are preferred, trastuzumab, tocilizumab, infliximab, bevacizumab, adalimumab, ustekinumab, golimumab or denosumab, more preferred tocilizumab, infliximab, infliximab, infliximab
  • tocilizumab is most preferred as the recombinant monoclonal antibody.
  • Tocilizumab has only to have the same amino acid sequence as the amino acid sequence (Gln1-Gly448) and L chain amino acid sequence (Asp1-Cys214) of Actemra (registered trademark), which are generally commercially available.
  • the amino acid sequence of Actemura H chain (Glu1-Gly448) and L chain amino acid sequence (Asp1-Cys214) are described in the sequence listing attached to International Publication No. 2005/090405.
  • 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 proline (Pro) instead of the 448 amino acid residue, or 449 amino acid residue in which lysine (Lys) is added to the 448th glycine (Gly). Also good.
  • the liquid preparation in the present invention contains 150 mg / mL to 200 mg / mL recombinant monoclonal antibody. Further, from the viewpoint that the effects of the present invention can be sufficiently exerted, it is preferable to contain 170 mg / mL to 190 mg / mL recombinant monoclonal antibody, and more preferably 180 mg / mL recombinant monoclonal antibody.
  • the recombinant monoclonal antibody used in the present invention is preferably not lyophilized or reconstituted in the production method.
  • the liquid preparation in the present invention contains an amino acid component of 45 mM to 94 mM having a histidine component of less than 5 mM.
  • the amino acid component other than the histidine component is not limited in type, but examples include arginine, arginine hydrochloride, methionine, glycine, phenylalanine, aspartic acid, glutamic acid, lysine, asparagine, tryptophan, cysteine and cysteine hydrochloride. Can be mentioned.
  • the amino acid component other than the histidine component is preferably at least one selected from the group consisting of arginine, arginine hydrochloride and methionine.
  • the amino acid component other than the histidine component preferably contains arginine, arginine hydrochloride and methionine, and more preferably contains arginine hydrochloride and methionine.
  • the histidine component is preferably at least one selected from the group consisting of histidine and histidine hydrochloride, and more preferably histidine and histidine hydrochloride.
  • the liquid preparation in the present invention may be any liquid preparation that contains less than 5 mM histidine, but preferably contains 1 mM or more and 4 mM or less histidine component from the viewpoint that the pH can be adjusted to a preferred range. More preferably, it contains a histidine component of 2 mM or more and 4 mM or less.
  • amino acid component it is preferable to contain an amino acid component of 45 mM to 94 mM in which the histidine component is less than 5 mM in the liquid preparation. Further, it preferably contains 75 mM to 93 mM amino acid components, more preferably 90 mM amino acid components.
  • the liquid preparation may further contain a polyol.
  • the polyol include propylene glycol, glycerin (glycerol), threose, threitol, erythrose, erythritol, ribose, arabinose, arabitol, lyxose, maltitol, sorbitol, sorbose, glucose, mannose, mannitol, levulose, dextrose, maltose, Examples include trehalose, fructose, xylitol, inositol, galactose, xylose, fructose, sucrose, and 1,2,6-hexanetriol.
  • sucrose As the polyol, sucrose, trehalose and the like are preferable, and sucrose is most preferable.
  • the liquid preparation may contain one or a combination of two or more of these polyols.
  • content of an amino acid component less than 70 mM, it is preferable to use combining a polyol from a viewpoint of dimer production
  • the content of the polyol is not particularly limited, and may be appropriately determined from the viewpoint of isotonicity of the liquid preparation. For example, it may be 30 mg / mL to 80 mg / mL, 40 mg / mL to 70 mg / mL, 50 mg / mL to 60 mg / mL.
  • the liquid formulation may contain other components necessary for formulating the liquid formulation.
  • a solubilizing agent for example, an isotonic agent, a preservative, an adsorption inhibitor, a sulfur-containing reducing agent, an antioxidant, preferably a solubilizing agent may be contained.
  • solubilizer examples include surfactants, particularly nonionic surfactants, specifically, polyoxyethylene hydrogenated castor oil, polysorbate (polysorbate 80, polysorbate 40, polysorbate 20, etc.), polyoxyethylene sorbitan mono Examples include laurate, castor oil fatty acid ethyl ester, and nicotinic acid amide.
  • isotonic agent examples include salts such as sodium chloride, potassium chloride, calcium chloride in addition to polyol.
  • preservative examples include methyl paraoxybenzoate, ethyl paraoxybenzoate, propyl paraoxybenzoate, sorbic acid, phenol, cresol, metacresol, and chlorocresol.
  • adsorption inhibitor examples include human serum albumin, lecithin, dextran, hydroxypropylcellulose, methylcellulose, and macrogol.
  • sulfur-containing reducing agent examples include N-acetylcysteine, N-acetylhomocysteine, thioctic acid, thiodiglycol, thioethanolamine, thioglycerol, thiosorbitol, thioglycolic acid and salts thereof, sodium thiosulfate, and glutathione. Can be mentioned.
  • antioxidants examples include erythorbic acid, dibutylhydroxytoluene, butylhydroxyanisole, ⁇ -tocopherol, tocopherol acetate, L-ascorbic acid and salts thereof, L-ascorbyl palmitate, L-ascorbic acid stearate, sodium bisulfite Sodium sulfite, triamyl gallate, propyl gallate, disodium ethylenediaminetetraacetate (EDTA ⁇ 2Na), sodium pyrophosphate, sodium metaphosphate.
  • the pH of the liquid formulation is 5.5 to 6.7.
  • the pH is preferably 5.8 to 6.4, more preferably 6.0 to 6.2, from the viewpoint of adjusting the kinematic viscosity of the liquid preparation to the extent that subcutaneous injection is possible.
  • the pH can be measured by, for example, a pH meter (model number: HM-30G, manufactured by Toa DKK Corporation).
  • the pH may be measured according to the method described in the 16th revised Japanese Pharmacopoeia, for example, at room temperature (15 ° C. to 25 ° C.).
  • the pH of a liquid formulation can be adjusted with the histidine component contained in a liquid formulation, the pH of a liquid formulation can also be adjusted using another buffer as needed.
  • buffering agents include, for example, phosphate (sodium or potassium), sodium bicarbonate, citrate (sodium or potassium), sodium acetate, sodium oxalate, phosphoric acid, carbonic acid, citric acid, succinic acid, apple Examples include acids, gluconic acid, and glycine.
  • the liquid preparation in the present invention can be administered, for example, by injection (subcutaneous injection, intravenous injection, intramuscular injection, etc.), transdermal, transmucosal, nasal, or transpulmonary.
  • subcutaneous injection is performed, the dose of recombinant monoclonal antibody per administration is as large as 150 mg / mL to 200 mg / mL, but the amount of injection solution is limited.
  • the liquid preparation in the present invention exhibits kinematic viscosity that enables subcutaneous injection even when it contains a recombinant monoclonal antibody at a high concentration of 150 mg / mL to 200 mg / mL.
  • the liquid preparation in the present invention is preferably used for subcutaneous injection from the viewpoint that the effects of the present invention can be achieved.
  • Subcutaneous injection is not only performed by doctors and other specialists as medical professionals, but may also be performed by the patient himself. Depending on the ethnic group and region, many patients are concerned about self-injection. It is preferable that the viscosity is low and the stability over time is excellent.
  • the kinematic viscosity of the liquid preparation in the present invention is preferably 6 mm 2 / s to 15 mm 2 / s, more preferably 7 mm 2 / s to 14 mm 2 / s, and more preferably 8 mm 2 / s to 12 mm 2 immediately after liquid preparation preparation. / S is more preferable.
  • the kinematic viscosity may be measured with an Ubbelohde viscometer (16th revision Japanese Pharmacopoeia General Test Method 2.53 Viscosity Measurement Method Method 1).
  • the dimer product and the low molecular weight decomposition product can be measured by size exclusion chromatography using, for example, a high performance liquid chromatograph system (Prominence, manufactured by Shimadzu Corporation).
  • the deamidated substance can be measured by ion exchange chromatography using, for example, a high performance liquid chromatograph system (Prominence, manufactured by Shimadzu Corporation).
  • Example 1 Confirmation of stabilization effect of liquid preparation
  • a liquid preparation containing a high concentration of recombinant monoclonal antibody (180 mg / mL as tocilizumab) contains 94 mM amino acid component and has a pH of 5.5 to 6.7. The effect on the stabilization of the formulation was evaluated.
  • 1-No. Five evaluation samples were prepared. The prescription of each evaluation sample is as follows. Note that “-” in the compositions in Tables 1 and 5 indicates not blended.
  • tocilizumab used in the examples should be prepared according to the methods described in International Publication No. 92/0197759, International Publication No. 2005/090405, International Publication No. 99/063058, and International Publication No. 2002/072615. Can do.
  • the evaluation sample was diluted with a mobile phase so that the protein concentration became 1 mg / mL, and used as an evaluation sample solution.
  • the evaluation results of the size exclusion chromatography method obtained in this example are shown in Table 2, the evaluation results of the ion exchange chromatography method are shown in Table 3, and the kinematic viscosity evaluation results are shown in Table 4.
  • the formulation containing 94 mM amino acid component and adjusted to pH 6 had a total amount of amino acids and a small content of histidine component, but at 60 ° C. for 2 weeks, 50 ° C.
  • the evaluation samples No. 1 and No. 2 in which the amount of the high molecular fraction and the low molecular fraction are large in both the total amount of amino acids and the content of the histidine component are used. It was comparable to 1.
  • the evaluation sample No. 5 shows that the increase in the acidic fraction containing the deamidated product in the ion-exchange chromatography method was evaluated sample No.
  • evaluation sample No. 2 adjusted to pH 5
  • evaluation sample No. 3, 3 and 4 adjusted to pH 6.8
  • evaluation sample No. of the polymer fraction containing the dimer Since an increasing tendency was observed as compared to 5, and an increasing tendency was also observed in the acidic fraction containing the deamidated substance, it was revealed that the pH range was an important management item. From the evaluation results of the formulation (evaluation sample No. 3 and 4) comparing the presence or absence of histidine addition, it is clear that the histidine component is an important component from the point of adjusting the pH of the liquid preparation to the expected pH range became. In addition, evaluation sample No. The kinematic viscosity of No.
  • Example 2 Confirmation of stability effect when content of amino acid component is further reduced Stabilizing effect for liquid preparation containing recombinant monoclonal antibody (180 mg / mL as tocilizumab) when content of amino acid component is further reduced below 94 mM Evaluated.
  • evaluation sample No. 6-1 to No. 8 was prepared. The prescription of each evaluation sample is as follows.
  • the evaluation results of the size exclusion chromatography method obtained in this example are shown in Table 6, the evaluation results of the ion exchange chromatography method are shown in Table 7, and the kinematic viscosity evaluation results are shown in Table 8.
  • each formulation was compared with the evaluation sample (evaluation sample No. 1) in which the amino acid component concentration shown in Example 1 had an amino acid content higher than 94 mM.
  • evaluation sample No. 1 the evaluation sample in which the amino acid component concentration shown in Example 1 had an amino acid content higher than 94 mM.
  • the preparation of the present invention is a stable high-concentration recombinant monoclonal antibody liquid preparation that realizes suppression of dimer formation and deamidation during long-term storage, and also exhibits high kinematic viscosity that enables subcutaneous injection Since a liquid preparation containing a recombinant monoclonal antibody can be provided at low cost, it is highly industrially useful.

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Abstract

The present invention can provide, at a low cost, a liquid formulation containing a high concentration of recombinant monoclonal antibodies comprising 150 mg/mL to 200 mg/mL of recombinant monoclonal antibodies, and 45 mM to 94 mM of an amino acid component in which the histidine component is less than 5 mM, and having a pH of 5.5 to 6.7. The liquid formulation containing a high concentration of recombinant monoclonal antibodies is stable, achieving limited dimer generation and limited deamidation during long-term storage, and exhibits a kinematic viscosity that allows for subcutaneous injection.

Description

高濃度抗体含有液体製剤High concentration antibody-containing liquid formulation
　本発明は、高濃度抗体含有液体製剤に関する。 The present invention relates to a liquid preparation containing a high concentration antibody.
　トシリズマブ等の組換えモノクローナル抗体は、現在、医薬品の有効成分等として利用されている。十分な治療効果を得るために、１回あたりの組換えモノクローナル抗体の投与量が大量となることがあるが、例えば自己注射が可能な皮下注射用製剤を使用するときには、１回の注射液量に制限がある。そのため、製剤中に含まれる組換えモノクローナル抗体濃度を高濃度化しなければならないことがある。 Recombinant monoclonal antibodies such as tocilizumab are currently used as active ingredients in pharmaceuticals. In order to obtain a sufficient therapeutic effect, the dose of the recombinant monoclonal antibody per administration may be large. For example, when using a preparation for subcutaneous injection capable of self-injection, the amount of injection once There are limitations. Therefore, it may be necessary to increase the concentration of the recombinant monoclonal antibody contained in the preparation.
　これまで、高濃度組換えモノクローナル抗体含有液体製剤として、特定量のアルギニン、メチオニン、ポリソルベートを含有する製剤処方（特許文献１）や、特定量のアルギニン塩酸塩、ヒスチジン、ポリソルベートを含有する製剤処方（特許文献２）等が知られている。
　なお、これらの製剤処方において、組換えモノクローナル抗体含有量が１５０ｍｇ／ｍＬ以上である場合には、アルギニン等のアミノ酸成分の含有量がいずれも１００ｍＭ以上となっている。 Until now, as a liquid preparation containing high-concentration recombinant monoclonal antibody, a pharmaceutical formulation containing a specific amount of arginine, methionine, polysorbate (Patent Document 1), or a pharmaceutical formulation containing a specific amount of arginine hydrochloride, histidine, polysorbate ( Patent Document 2) and the like are known.
In these pharmaceutical formulations, when the content of the recombinant monoclonal antibody is 150 mg / mL or more, the content of amino acid components such as arginine is 100 mM or more.
国際公開第２００９／０８４６５９号International Publication No. 2009/084659 国際公開第２００４／０９１６５８号International Publication No. 2004/091658
　従来、高濃度の組換えモノクローナル抗体を含有する液体製剤を調製するためには、１００ｍＭ以上という高濃度のアミノ酸成分を添加する必要があると考えられてきた。そのため、長期保存時の二量体生成抑制及び脱アミド化抑制を実現し、且つ、皮下注射を可能にする動粘度を示す高濃度組換えモノクローナル抗体含有液体製剤を低コストで提供するためには、更なる技術開発が必要である。 Conventionally, in order to prepare a liquid preparation containing a high concentration of recombinant monoclonal antibody, it has been considered necessary to add a high concentration amino acid component of 100 mM or more. Therefore, in order to provide a liquid preparation containing a high-concentration recombinant monoclonal antibody that realizes suppression of dimer formation and suppression of deamidation during long-term storage and exhibits kinematic viscosity that enables subcutaneous injection at low cost. Further technical development is necessary.
　本発明は、長期保存時の二量体生成抑制及び脱アミド化抑制を実現した安定な高濃度組換えモノクローナル抗体液体製剤であり、且つ、皮下注射を可能にする動粘度を示す高濃度組換えモノクローナル抗体含有液体製剤を低コストで提供することを課題とする。 The present invention is a stable high-concentration recombinant monoclonal antibody liquid preparation that achieves dimer formation suppression and deamidation suppression during long-term storage, and also exhibits high-concentration recombination that exhibits kinematic viscosity that enables subcutaneous injection An object is to provide a liquid preparation containing a monoclonal antibody at low cost.
　本発明は以下のとおりである。
＜１＞　１５０ｍｇ／ｍＬ～２００ｍｇ／ｍＬの組換えモノクローナル抗体と、ヒスチジン成分が５ｍＭ未満である４５ｍＭ～９４ｍＭのアミノ酸成分とを含有し、ｐＨが５．５～６．７である高濃度組換えモノクローナル抗体含有液体製剤。
＜２＞　ヒスチジン成分以外のアミノ酸成分が、アルギニン、アルギニン塩酸塩及びメチオニンからなる群より選ばれる少なくとも１種である＜１＞に記載の液体製剤。
＜３＞　ヒスチジン成分が、ヒスチジン及びヒスチジン塩酸塩からなる群より選ばれる少なくとも１種である＜１＞又は＜２＞に記載の液体製剤。
＜４＞　７５ｍＭ～９３ｍＭのアミノ酸成分を含有する＜１＞ないし＜３＞のいずれか１つに記載の液体製剤。
＜５＞　９０ｍＭのアミノ酸成分を含有する＜１＞ないし＜４＞のいずれか１つに記載の液体製剤。
＜６＞　ｐＨが５．８～６．４である＜１＞ないし＜５＞のいずれか１つに記載の液体製剤。
＜７＞　ｐＨが６．０～６．２である＜１＞ないし＜６＞のいずれか１つに記載の液体製剤。
＜８＞　１７０ｍｇ／ｍＬ～１９０ｍｇ／ｍＬの組換えモノクローナル抗体を含有する＜１＞ないし＜７＞のいずれか１つに記載の液体製剤。
＜９＞　１８０ｍｇ／ｍＬの組換えモノクローナル抗体を含有する＜１＞ないし＜８＞のいずれか１つに記載の液体製剤。
＜１０＞　さらにポリオールを含有する＜１＞ないし＜９＞のいずれか１つに記載の液体製剤。
＜１１＞　組換えモノクローナル抗体が、トシリズマブ、トラスツズマブ、リツキシマブ、パリビズマブ、インフリキシマブ、バシリキシマブ、ゲムツズマブオゾガマイシン、ベバシズマブ、イブリツモマブ　チウキセタン、アダリムマブ、セツキシマブ、ラニビズマブ、オマリズマブ、エクリズマブ、パニツムマブ、ウステキヌマブ、ゴリムマブ、カナキヌマブ、デノスマブ、モガムリズマブ、オファツムマブ、ペルツズマブ、トラスツズマブエムタンシン、ブレンツキシマブ　ベドチン、ナタリズマブ、ニボルマブ、アレムツズマブ、セクキヌマブ、ラムシルマブ及びイピリムマブからなる群から選択される少なくとも一つである＜１０＞に記載の液体製剤。 The present invention is as follows.
<1> High-concentration recombination containing 150 mg / mL to 200 mg / mL recombinant monoclonal antibody and 45 mM to 94 mM amino acid component having a histidine component of less than 5 mM and a pH of 5.5 to 6.7 Monoclonal antibody-containing liquid preparation.
<2> The liquid preparation according to <1>, 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.
<3> The liquid formulation according to <1> or <2>, wherein the histidine component is at least one selected from the group consisting of histidine and histidine hydrochloride.
<4> The liquid preparation according to any one of <1> to <3>, comprising an amino acid component of 75 mM to 93 mM.
<5> The liquid preparation according to any one of <1> to <4>, containing 90 mM amino acid component.
<6> The liquid preparation according to any one of <1> to <5>, wherein the pH is 5.8 to 6.4.
<7> The liquid preparation according to any one of <1> to <6>, wherein the pH is 6.0 to 6.2.
<8> The liquid preparation according to any one of <1> to <7>, comprising 170 mg / mL to 190 mg / mL recombinant monoclonal antibody.
<9> The liquid preparation according to any one of <1> to <8>, which contains 180 mg / mL recombinant monoclonal antibody.
<10> The liquid preparation according to any one of <1> to <9>, further containing a polyol.
<11> Recombinant monoclonal antibodies are tocilizumab, trastuzumab, rituximab, rituximab, rituximab, rituximab, rituximab, rituximab, rituximab, rituximab, rituximab, rituximab, rituximab, rituximab, rituximab, rituximab At least one liquid selected from the group consisting of canakinumab, denosumab, mogamulizumab, ofatumumab, pertuzumab, trastuzumab emtansine, brentuximab vedotin, natalizumab, nivolumab, alemtuzumab, secukinumab, ramucimab and ipilimumab .
　本発明によれば、長期保存時の二量体生成抑制及び脱アミド化抑制を実現した安定な高濃度組換えモノクローナル抗体液体製剤であり、且つ、皮下注射を可能にする動粘度を示す高濃度組換えモノクローナル抗体含有液体製剤を低コストで提供することができる。 According to the present invention, it is a stable high-concentration recombinant monoclonal antibody liquid preparation that realizes dimer formation inhibition and deamidation inhibition during long-term storage, and has a high concentration that exhibits kinematic viscosity that enables subcutaneous injection. A liquid preparation containing a recombinant monoclonal antibody can be provided at low cost.
　以下、本発明について詳細に説明する。
　本発明における高濃度組換えモノクローナル抗体含有液体製剤（以下、「液体製剤」とも称する。）は、１５０ｍｇ／ｍＬ～２００ｍｇ／ｍＬの組換えモノクローナル抗体と、ヒスチジン成分が５ｍＭ未満である４５ｍＭ～９４ｍＭのアミノ酸成分（合計）とを含有し、ｐＨが５．５～６．７である。また、液体製剤は、他の成分を含んでいてもよい。
　本発明における液体製剤は、ヒスチジン成分が５ｍＭ未満である４５ｍＭ～９４ｍＭのアミノ酸成分を含有し、ｐＨを５．５～６．７の範囲に調整することにより、１５０ｍｇ／ｍＬ～２００ｍｇ／ｍＬもの高濃度の組換えモノクローナル抗体を含有しても、液体製剤中での、長期保存時の二量体生成抑制及び脱アミド化抑制を実現し、且つ、皮下注射を可能にする動粘度を示すという効果を奏し得る。さらに、本発明における液体製剤は、４５ｍＭ～９４ｍＭのアミノ酸成分という低用量のアミノ酸成分を含有するため、従来の高濃度組換えモノクローナル抗体含有液体製剤に比べて低コストで提供することができる。
　なお、本明細書において「～」を用いて示された数値範囲は、「～」の前後に記載される数値をそれぞれ最小値及び最大値として含む範囲を示す。
　また、モル濃度の単位は、Ｍ（モーラー）で、ｍＭは１０－３ｍｏｌ／Ｌである。
　本明細書において、長期保存とは、例えば２℃～８℃で１年以上保存することが挙げられ、好ましくは２℃～８℃で２年以上保存することが挙げられ、より好ましくは、２℃～８℃で２年～３年保存することが挙げられる。 Hereinafter, the present invention will be described in detail.
A liquid preparation containing a high-concentration recombinant monoclonal antibody in the present invention (hereinafter also referred to as “liquid preparation”) comprises a recombinant monoclonal antibody of 150 mg / mL to 200 mg / mL and 45 mM to 94 mM having a histidine component of less than 5 mM. The amino acid components (total) are contained, and the pH is 5.5 to 6.7. The liquid preparation may contain other components.
The liquid preparation in the present invention contains an amino acid component of 45 mM to 94 mM in which the histidine component is less than 5 mM, and the pH is adjusted to the range of 5.5 to 6.7, and as high as 150 mg / mL to 200 mg / mL. Even if it contains a recombinant monoclonal antibody at a concentration, it has the effect of suppressing dimer formation and deamidation during long-term storage in liquid formulations, and exhibiting kinematic viscosity that enables subcutaneous injection Can be played. Furthermore, since the liquid preparation in the present invention contains a low-dose amino acid component of 45 mM to 94 mM amino acid component, it can be provided at a lower cost than conventional liquid preparations containing a high concentration recombinant monoclonal antibody.
In the present specification, a numerical range indicated by using “to” indicates a range including the numerical values described before and after “to” as the minimum value and the maximum value, respectively.
The unit of molar concentration is M (molar), and mM is 10 −3 mol / L.
In the present specification, long-term storage includes, for example, storage at 2 ° C. to 8 ° C. for 1 year or longer, preferably storage at 2 ° C. to 8 ° C. for 2 years or longer, more preferably 2 It can be stored for 2 to 3 years at -8 to 8 ° C.
（組換えモノクローナル抗体）
　組換えモノクローナル抗体とは、組換えＤＮＡ技術を応用して形質転換された細胞により生産される抗体である。組換えモノクローナル抗体は、動物細胞で発現又は分泌されるものであれば好ましいが、組換えモノクローナル抗体の種類が特に制限されるものではない。例えば医薬品として使用可能な組換えモノクローナル抗体であることが好ましい。 (Recombinant monoclonal antibody)
A recombinant monoclonal antibody is an antibody produced by cells transformed by applying recombinant DNA technology. The recombinant monoclonal antibody is preferable if 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 that can be used as a pharmaceutical is preferable.
　なお、本発明における液体製剤に含有し得る組換えモノクローナル抗体は、ヒト、マウス、ラット等の動物由来の組換えモノクローナル抗体だけではなく、キメラ抗体、ヒト化抗体等の組換えモノクローナル抗体も含まれる。また、抗体の免疫グロブリンクラスは特に限定されるものではなく、ＩｇＧ１、ＩｇＧ２、ＩｇＧ３、ＩｇＧ４などのＩｇＧ、ＩｇＡ、ＩｇＤ、ＩｇＥ、ＩｇＭなどいずれのクラスでもよい。 Recombinant monoclonal antibodies that can be contained in the liquid preparation of the present invention include not only recombinant monoclonal antibodies derived from animals such as humans, mice, and rats, but also recombinant monoclonal antibodies such as chimeric antibodies and humanized antibodies. . The immunoglobulin class of the antibody is not particularly limited, and any class such as IgG such as IgG1, IgG2, IgG3, and IgG4, IgA, IgD, IgE, and IgM may be used.
　また、組換えモノクローナル抗体には、Ｆｖ、Ｆａｂ、Ｆ（ａｂ）２などの抗体断片や、抗体の可変領域をペプチドリンカー等のリンカーで結合させた１価または２価以上の一本鎖Ｆｖ（ｓｃＦｖ、ｓｃ（Ｆｖ）２やｓｃＦｖダイマーなどのＤｉａｂｏｄｙ等）などの低分子化抗体なども含まれる。
　これらの組換えモノクローナル抗体は、国際公開第９２／０１９７５９号及び国際公開第２００５／０９０４０５号に記載の方法に準じ調製することができる。 Recombinant monoclonal antibodies include antibody fragments such as Fv, Fab, F (ab) 2, and monovalent or bivalent or more single-chain Fvs (variable regions of antibodies bound by a linker such as a peptide linker). scFv, sc (Fv) 2 , Diabodies such as scFv dimer) and the like are also included.
These recombinant monoclonal antibodies can be prepared according to the methods described in International Publication No. 92/019759 and International Publication No. 2005/090405.
　組換えモノクローナル抗体としては、例えば、その種類が限定されるものではないが、例えば、トラスツズマブ、リツキシマブ、パリビズマブ、インフリキシマブ、バシリキシマブ、ゲムツズマブオゾガマイシン、ベバシズマブ、イブリツモマブ　チウキセタン、トシリズマブ、アダリムマブ、セツキシマブ、ラニビズマブ、オマリズマブ、エクリズマブ、パニツムマブ、ウステキヌマブ、ゴリムマブ、カナキヌマブ、デノスマブ、モガムリズマブ、オファツムマブ、ペルツズマブ、トラスツズマブ　エムタンシン、ブレンツキシマブ　ベドチン、ナタリズマブ、ニボルマブ、アレムツズマブ、セクキヌマブ、ラムシルマブ及びイピリムマブが挙げられる。 Examples of the recombinant monoclonal antibody include, but are not limited to, for example, trastuzumab, rituximab, palivizumab, infliximab, basiliximab, gemtuzumab ozogamicin, bevacizumab, ibritumomab tiuxetane, tocilizumab, adalimumab, , Ranibizumab, Omalizumab, Eculizumab, Panitumumab, Usutekinumab, Golimumab, Kanakinumab, Denosumab, Mogamulizumab, Offatumumab, Pertuzumab, Trastuzumab Emtansin, Brentuximab, Beduminumab
　また、組換えモノクローナル抗体としては、トシリズマブ、トラスツズマブ、リツキシマブ、パリビズマブ、インフリキシマブ、バシリキシマブ、ベバシズマブ、アダリムマブ、セツキシマブ、ラニビズマブ、オマリズマブ、エクリズマブ、パニツムマブ、ウステキヌマブ、ゴリムマブ、カナキヌマブ、デノスマブ、モガムリズマブ、オファツムマブ、ペルツズマブ、ナタリズマブ、ニボルマブ、アレムツズマブ、セクキヌマブ、ラムシルマブ又はイピリムマブが好ましく、トラスツズマブ、トシリズマブ、インフリキシマブ、ベバシズマブ、アダリムマブ、ウステキヌマブ、ゴリムマブ又はデノスマブが好ましく、トシリズマブ、インフリキシマブ、ベバシズマブ、アダリムマブ又はデノスマブがより好ましい。 Recombinant monoclonal antibodies include tocilizumab, trastuzumab, rituximab, paclitumumab, infliximab, infliximab, basiliximab, bevacizumab, adalimumab, cetuximab, ranibizumab, omalizumab, eculizumab, panitumabum , Nivolumab, alemtuzumab, secukinumab, ramcilumab or ipilimumab are preferred, trastuzumab, tocilizumab, infliximab, bevacizumab, adalimumab, ustekinumab, golimumab or denosumab, more preferred tocilizumab, infliximab, infliximab, infliximab
　さらに、組換えモノクローナル抗体としてはトシリズマブが最も好ましい。
　トシリズマブは、一般に市販されているアクテムラ（登録商標）のＨ鎖のアミノ酸配列（Ｇｌｎ１－Ｇｌｙ４４８）及びＬ鎖のアミノ酸配列（Ａｓｐ１－Ｃｙｓ２１４）と、同一のアミノ酸配列を有するものであればよい。なお、アクテムラのＨ鎖のアミノ酸配列（Ｇｌｕ１－Ｇｌｙ４４８）及びＬ鎖のアミノ酸配列（Ａｓｐ１－Ｃｙｓ２１４）は、国際公開第２００５／０９０４０５号に添付の配列表に記載されている。
　また、Ｈ鎖のＮ末端残基は、グルタミン酸である代わりに、ピログルタミン酸（Ｐｙｒｏｇｌｕｔａｍｉｃ　ａｃｉｄ；ｐＧｌｕ）であってもよい。Ｈ鎖のＣ末端残基は４４８アミノ酸残基の代わりに、４４７のプロリン（Ｐｒｏ）まででもよいし、４４８番目のグリシン（Ｇｌｙ）にリシン（Ｌｙｓ）が付加された４４９アミノ酸残基であってもよい。 Furthermore, tocilizumab is most preferred as the recombinant monoclonal antibody.
Tocilizumab has only to have the same amino acid sequence as the amino acid sequence (Gln1-Gly448) and L chain amino acid sequence (Asp1-Cys214) of Actemra (registered trademark), which are generally commercially available. The amino acid sequence of Actemura H chain (Glu1-Gly448) and L chain amino acid sequence (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 proline (Pro) instead of the 448 amino acid residue, or 449 amino acid residue in which lysine (Lys) is added to the 448th glycine (Gly). Also good.
　本発明における液体製剤は、１５０ｍｇ／ｍＬ～２００ｍｇ／ｍＬの組換えモノクローナル抗体を含有する。また、本発明の効果を十分に奏し得るという観点から、１７０ｍｇ／ｍＬ～１９０ｍｇ／ｍＬの組換えモノクローナル抗体を含有することが好ましく、１８０ｍｇ／ｍＬの組換えモノクローナル抗体を含有することがより好ましい。本発明に用いる組換えモノクローナル抗体は、その製造方法において凍結乾燥、再構成は行わないのが好ましい。 The liquid preparation in the present invention contains 150 mg / mL to 200 mg / mL recombinant monoclonal antibody. Further, from the viewpoint that the effects of the present invention can be sufficiently exerted, it is preferable to contain 170 mg / mL to 190 mg / mL recombinant monoclonal antibody, and more preferably 180 mg / mL recombinant monoclonal antibody. The recombinant monoclonal antibody used in the present invention is preferably not lyophilized or reconstituted in the production method.
（アミノ酸成分）
　本発明における液体製剤は、ヒスチジン成分が５ｍＭ未満である４５ｍＭ～９４ｍＭのアミノ酸成分を含有する。
　ヒスチジン成分以外のアミノ酸成分としては、種類が限定されるものではないが、例えば、アルギニン、アルギニン塩酸塩、メチオニン、グリシン、フェニルアラニン、アスパラギン酸、グルタミン酸、リジン、アスパラギン、トリプトファン、システイン及びシステイン塩酸塩が挙げられる。また、長期保存時の二量体生成抑制及び脱アミド化抑制の観点より、ヒスチジン成分以外のアミノ酸成分としては、アルギニン、アルギニン塩酸塩及びメチオニンからなる群より選ばれる少なくとも１種であることが好ましい。さらに、ヒスチジン成分以外のアミノ酸成分としては、アルギニン、アルギニン塩酸塩及びメチオニンを含有することが好ましく、アルギニン塩酸塩及びメチオニンを含有することがより好ましい。 (Amino acid component)
The liquid preparation in the present invention contains an amino acid component of 45 mM to 94 mM having a histidine component of less than 5 mM.
The amino acid component other than the histidine component is not limited in type, but examples include arginine, arginine hydrochloride, methionine, glycine, phenylalanine, aspartic acid, glutamic acid, lysine, asparagine, tryptophan, cysteine and cysteine hydrochloride. Can be mentioned. Further, from the viewpoint of suppressing dimer formation and deamidation during long-term storage, the amino acid component other than the histidine component is preferably at least one selected from the group consisting of arginine, arginine hydrochloride and methionine. . Furthermore, the amino acid component other than the histidine component preferably contains arginine, arginine hydrochloride and methionine, and more preferably contains arginine hydrochloride and methionine.
　ヒスチジン成分としては、ヒスチジン及びヒスチジン塩酸塩からなる群より選ばれる少なくとも１種であることが好ましく、ヒスチジン及びヒスチジン塩酸塩であることがより好ましい。
　本発明における液体製剤は、液体製剤中に５ｍＭ未満のヒスチジンを含有するものであればよいが、ｐＨを好ましい範囲に調整し得るという観点より、１ｍＭ以上４ｍＭ以下のヒスチジン成分を含有することが好ましく、２ｍＭ以上４ｍＭ以下のヒスチジン成分を含有することがより好ましい。 The histidine component is preferably at least one selected from the group consisting of histidine and histidine hydrochloride, and more preferably histidine and histidine hydrochloride.
The liquid preparation in the present invention may be any liquid preparation that contains less than 5 mM histidine, but preferably contains 1 mM or more and 4 mM or less histidine component from the viewpoint that the pH can be adjusted to a preferred range. More preferably, it contains a histidine component of 2 mM or more and 4 mM or less.
　アミノ酸成分としては、ヒスチジン成分が５ｍＭ未満である４５ｍＭ～９４ｍＭのアミノ酸成分を液体製剤中に含有することが好ましい。また、７５ｍＭ～９３ｍＭのアミノ酸成分を含有することが好ましく、９０ｍＭのアミノ酸成分を含有することがより好ましい。 As the amino acid component, it is preferable to contain an amino acid component of 45 mM to 94 mM in which the histidine component is less than 5 mM in the liquid preparation. Further, it preferably contains 75 mM to 93 mM amino acid components, more preferably 90 mM amino acid components.
（ポリオール）
　液体製剤は、さらに、ポリオールを含有していてもよい。
　ポリオールとしては、例えば、プロピレングリコール、グリセリン（グリセロール）、トレオース、トレイトール、エリトロース、エリトリトール、リボース、アラビノース、アラビトール、リキソース、マルチトール、ソルビトール、ソルボース、グルコース、マンノース、マンニトール、レブロース、デキストロース、マルトース、トレハロース、フルクトース、キシリトール、イノシトール、ガラクトース、キシロース、フルクトース、スクロース、１，２，６－ヘキサントリオールが挙げられる。中でも、ポリオールとしては、スクロース、トレハロース等が好ましく、スクロースが最も好ましい。
　液体製剤は、これらのポリオールの１種又は２種以上を組合せて含有していてもよい。
　また、アミノ酸成分の含有量を７０ｍＭ未満にする場合には、二量体生成抑制の観点より、ポリオールを組合せて用いることが好ましい。 (Polyol)
The liquid preparation may further contain a polyol.
Examples of the polyol include propylene glycol, glycerin (glycerol), threose, threitol, erythrose, erythritol, ribose, arabinose, arabitol, lyxose, maltitol, sorbitol, sorbose, glucose, mannose, mannitol, levulose, dextrose, maltose, Examples include trehalose, fructose, xylitol, inositol, galactose, xylose, fructose, sucrose, and 1,2,6-hexanetriol. Among these, as the polyol, sucrose, trehalose and the like are preferable, and sucrose is most preferable.
The liquid preparation may contain one or a combination of two or more of these polyols.
Moreover, when making content of an amino acid component less than 70 mM, it is preferable to use combining a polyol from a viewpoint of dimer production | generation suppression.
　ポリオールの含有量は特に限定されるものではなく、液体製剤等張化の観点より、適宜決定すればよい。例えば、３０ｍｇ／ｍＬ～８０ｍｇ／ｍＬ、４０ｍｇ／ｍＬ～７０ｍｇ／ｍＬ、５０ｍｇ／ｍＬ～６０ｍｇ／ｍＬであればよい。 The content of the polyol is not particularly limited, and may be appropriately determined from the viewpoint of isotonicity of the liquid preparation. For example, it may be 30 mg / mL to 80 mg / mL, 40 mg / mL to 70 mg / mL, 50 mg / mL to 60 mg / mL.
（その他成分）
　液体製剤は、組換えモノクローナル抗体及びアミノ酸成分の他に、液体製剤の製剤化に必要な他の成分を含有していてもよい。他の成分としては、例えば、溶解補助剤、等張化剤、保存剤、吸着防止剤、含硫還元剤、酸化防止剤、好ましくは溶解補助剤を含有していてもよい。 (Other ingredients)
In addition to the recombinant monoclonal antibody and amino acid component, the liquid formulation may contain other components necessary for formulating the liquid formulation. As other components, for example, a solubilizing agent, an isotonic agent, a preservative, an adsorption inhibitor, a sulfur-containing reducing agent, an antioxidant, preferably a solubilizing agent may be contained.
　溶解補助剤としては、例えば、界面活性剤、特に非イオン性界面活性剤、具体的には、ポリオキシエチレン硬化ヒマシ油、ポリソルベート（ポリソルベート８０、ポリソルベート４０、ポリソルベート２０等）、ポリオキシエチレンソルビタンモノラウレート、ヒマシ油脂肪酸エチルエステル、及びニコチン酸アミド等が挙げられる。 Examples of the solubilizer include surfactants, particularly nonionic surfactants, specifically, polyoxyethylene hydrogenated castor oil, polysorbate (polysorbate 80, polysorbate 40, polysorbate 20, etc.), polyoxyethylene sorbitan mono Examples include laurate, castor oil fatty acid ethyl ester, and nicotinic acid amide.
　等張化剤としては、例えば、ポリオールの他、塩化ナトリウム、塩化カリウム、塩化カルシウム等の塩類が挙げられる。 Examples of the isotonic agent include salts such as sodium chloride, potassium chloride, calcium chloride in addition to polyol.
　保存剤としては、例えば、パラオキシ安息香酸メチル、パラオキシ安息香酸エチル、パラオキシ安息香酸プロピル、ソルビン酸、フェノール、クレゾール、メタクレゾール、クロロクレゾールが挙げられる。 Examples of the preservative include methyl paraoxybenzoate, ethyl paraoxybenzoate, propyl paraoxybenzoate, sorbic acid, phenol, cresol, metacresol, and chlorocresol.
　吸着防止剤としては、例えば、ヒト血清アルブミン、レシチン、デキストラン、ヒドロキシプロピルセルロース、メチルセルロース、マクロゴールが挙げられる。 Examples of the adsorption inhibitor include human serum albumin, lecithin, dextran, hydroxypropylcellulose, methylcellulose, and macrogol.
　含硫還元剤としては、例えば、Ｎ－アセチルシステイン、Ｎ－アセチルホモシステイン、チオクト酸、チオジグリコール、チオエタノールアミン、チオグリセロール、チオソルビトール、チオグリコール酸及びその塩、チオ硫酸ナトリウム、グルタチオンが挙げられる。 Examples of the sulfur-containing reducing agent include N-acetylcysteine, N-acetylhomocysteine, thioctic acid, thiodiglycol, thioethanolamine, thioglycerol, thiosorbitol, thioglycolic acid and salts thereof, sodium thiosulfate, and glutathione. Can be mentioned.
　酸化防止剤としては、例えば、エリソルビン酸、ジブチルヒドロキシトルエン、ブチルヒドロキシアニソール、α－トコフェロール、酢酸トコフェロール、Ｌ－アスコルビン酸及びその塩、Ｌ－アスコルビン酸パルミテート、Ｌ－アスコルビン酸ステアレート、亜硫酸水素ナトリウム、亜硫酸ナトリウム、没食子酸トリアミル、没食子酸プロピル、エチレンジアミン四酢酸二ナトリウム（ＥＤＴＡ・２Ｎａ）、ピロリン酸ナトリウム、メタリン酸ナトリウムが挙げられる。 Examples of the antioxidant include erythorbic acid, dibutylhydroxytoluene, butylhydroxyanisole, α-tocopherol, tocopherol acetate, L-ascorbic acid and salts thereof, L-ascorbyl palmitate, L-ascorbic acid stearate, sodium bisulfite Sodium sulfite, triamyl gallate, propyl gallate, disodium ethylenediaminetetraacetate (EDTA · 2Na), sodium pyrophosphate, sodium metaphosphate.
　なお、１５０ｍｇ／ｍＬ以下、例えば１００ｍｇ／ｍＬの組換えモノクローナル抗体を含有する組換えモノクローナル抗体含有液体製剤を調製する際に、ヒスチジン成分が５ｍＭ未満である４５ｍＭ～９４ｍＭのアミノ酸成分を含有し、ｐＨが５．５～６．７である組換えモノクローナル抗体含有液体製剤を調製することもできる。 When preparing a recombinant monoclonal antibody-containing liquid preparation containing a recombinant monoclonal antibody of 150 mg / mL or less, for example, 100 mg / mL, a 45 mM to 94 mM amino acid component having a histidine component of less than 5 mM is contained, and the pH It is also possible to prepare a liquid preparation containing a recombinant monoclonal antibody having a pH of 5.5 to 6.7.
（ｐＨ）
　液体製剤のｐＨは、５．５～６．７である。また、皮下注射が可能な範囲に、液体製剤の動粘度を調整するという観点より、ｐＨは５．８～６．４であることが好ましく、６．０～６．２であることがより好ましい。ｐＨの測定は、例えば、ｐＨメータ（型番：ＨＭ－３０Ｇ、東亜ディーケーケー（株）製）により行うことができる。また、ｐＨの測定は第１６改正日本薬局方に記載の方法に従い測定し、例えば常温（１５℃～２５℃）で測定すればよい。
　液体製剤のｐＨは、液体製剤に含有されるヒスチジン成分により調整され得るものであるが、必要に応じて、他の緩衝剤を使用して液体製剤のｐＨを調整することもできる。他の緩衝剤としては、例えば、リン酸塩（ナトリウム又はカリウム）、炭酸水素ナトリウム、クエン酸塩（ナトリウム又はカリウム）、酢酸ナトリウム、琥珀酸ナトリウム、リン酸、炭酸、クエン酸、琥珀酸、リンゴ酸、グルコン酸、グリシンが挙げられる。 (PH)
The pH of the liquid formulation is 5.5 to 6.7. In addition, the pH is preferably 5.8 to 6.4, more preferably 6.0 to 6.2, from the viewpoint of adjusting the kinematic viscosity of the liquid preparation to the extent that subcutaneous injection is possible. . The pH can be measured by, for example, a pH meter (model number: HM-30G, manufactured by Toa DKK Corporation). The pH may be measured according to the method described in the 16th revised Japanese Pharmacopoeia, for example, at room temperature (15 ° C. to 25 ° C.).
Although the pH of a liquid formulation can be adjusted with the histidine component contained in a liquid formulation, the pH of a liquid formulation can also be adjusted using another buffer as needed. Other buffering agents include, for example, phosphate (sodium or potassium), sodium bicarbonate, citrate (sodium or potassium), sodium acetate, sodium oxalate, phosphoric acid, carbonic acid, citric acid, succinic acid, apple Examples include acids, gluconic acid, and glycine.
　本発明における液体製剤は、例えば注射剤（皮下注射、静脈注射、筋肉注射等）、経皮、経粘膜、経鼻、経肺で投与することができる。
　皮下注射を行うときには、１回あたりの組換えモノクローナル抗体投与量が１５０ｍｇ／ｍＬ～２００ｍｇ／ｍＬ等と大量となる一方で、注射液量の制限がある。本発明における液体製剤は、１５０ｍｇ／ｍＬ～２００ｍｇ／ｍＬという高濃度の組換えモノクローナル抗体を含有しても、皮下注射を可能にする動粘度を示すものである。そのため、本発明の効果を奏し得るという点より、本発明における液体製剤は、皮下注射用として用いることが好ましい。皮下注射は医者をはじめとする医療従事者としての専門家が行う場合だけでなく、患者本人が行う自己注射である場合があり、民族、地域によっては、自己注射に不安を持つ患者も多く動粘度が低く、その経時の安定性が優れていることは好ましい。 The liquid preparation in the present invention can be administered, for example, by injection (subcutaneous injection, intravenous injection, intramuscular injection, etc.), transdermal, transmucosal, nasal, or transpulmonary.
When subcutaneous injection is performed, the dose of recombinant monoclonal antibody per administration is as large as 150 mg / mL to 200 mg / mL, but the amount of injection solution is limited. The liquid preparation in the present invention exhibits kinematic viscosity that enables subcutaneous injection even when it contains a recombinant monoclonal antibody at a high concentration of 150 mg / mL to 200 mg / mL. Therefore, the liquid preparation in the present invention is preferably used for subcutaneous injection from the viewpoint that the effects of the present invention can be achieved. Subcutaneous injection is not only performed by doctors and other specialists as medical professionals, but may also be performed by the patient himself. Depending on the ethnic group and region, many patients are worried about self-injection. It is preferable that the viscosity is low and the stability over time is excellent.
　本発明における液体製剤の動粘度は、例えば、液体製剤調整直後において、６ｍｍ２／ｓ～１５ｍｍ２／ｓが好ましく、７ｍｍ２／ｓ～１４ｍｍ２／ｓがより好ましく、８ｍｍ２／ｓ～１２ｍｍ２／ｓがより好ましい。
　動粘度は、ウベローデ型粘度計（第１６改正　日本薬局方　一般試験法　２．５３　粘度測定法　第１法）で測定すればよい。
　二量体生成物及び低分子量分解物は、例えば高速液体クロマトグラフシステム（Ｐｒｏｍｉｎｅｎｃｅ、株式会社島津製作所製）を用いて、サイズ排除クロマトグラフィ法により測定することができる。
　脱アミド体は、例えば高速液体クロマトグラフシステム（Ｐｒｏｍｉｎｅｎｃｅ、株式会社島津製作所製）を用いて、イオン交換クロマトグラフ法により測定することができる。 The kinematic viscosity of the liquid preparation in the present invention is preferably 6 mm 2 / s to 15 mm 2 / s, more preferably 7 mm 2 / s to 14 mm 2 / s, and more preferably 8 mm 2 / s to 12 mm 2 immediately after liquid preparation preparation. / S is more preferable.
The kinematic viscosity may be measured with an Ubbelohde viscometer (16th revision Japanese Pharmacopoeia General Test Method 2.53 Viscosity Measurement Method Method 1).
The dimer product and the low molecular weight decomposition product can be measured by size exclusion chromatography using, for example, a high performance liquid chromatograph system (Prominence, manufactured by Shimadzu Corporation).
The deamidated substance can be measured by ion exchange chromatography using, for example, a high performance liquid chromatograph system (Prominence, manufactured by Shimadzu Corporation).
　以下の実施例により本発明を更に詳述するが、本発明はこれら実施例に限定して理解されるべきものではない。本明細書で、％の表示は特に記載しない限り質量％である。 The present invention will be described in further detail with reference to the following examples, but the present invention should not be construed as being limited to these examples. In this specification, the indication of% is mass% unless otherwise specified.
（実施例１）
液体製剤の安定化効果の確認
　高濃度の組換えモノクローナル抗体（トシリズマブとして１８０ｍｇ／ｍＬ）を含む液体製剤について、９４ｍＭのアミノ酸成分を含有し、ｐＨが５．５～６．７である本発明の製剤の安定化に及ぼす影響を評価した。
　本検討では、液体製剤の安定化効果を確認するために、Ｎｏ．１～Ｎｏ．５の評価試料を調製した。各評価試料の処方は以下の通りである。なお、表１及び表５中の組成における「－」は未配合を示す。また、実施例で使用したトシリズマブは、国際公開第９２／０１９７５９号、国際公開第２００５／０９０４０５号、国際公開第９９／０６３０５８号及び国際公開第２００２／０７２６１５号に記載の方法に準じ調製することができる。 Example 1
Confirmation of stabilization effect of liquid preparation A liquid preparation containing a high concentration of recombinant monoclonal antibody (180 mg / mL as tocilizumab) contains 94 mM amino acid component and has a pH of 5.5 to 6.7. The effect on the stabilization of the formulation was evaluated.
In this study, in order to confirm the stabilizing effect of the liquid preparation, 1-No. Five evaluation samples were prepared. The prescription of each evaluation sample is as follows. Note that “-” in the compositions in Tables 1 and 5 indicates not blended. In addition, tocilizumab used in the examples should be prepared according to the methods described in International Publication No. 92/0197759, International Publication No. 2005/090405, International Publication No. 99/063058, and International Publication No. 2002/072615. Can do.
Figure JPOXMLDOC01-appb-T000001
Figure JPOXMLDOC01-appb-T000001
　液体製剤の安定性を評価するために、各評価試料を２ｍＬガラスバイアルに０．５ｍＬ充填して、各評価試料の熱加速試験（６０℃－２週間、５０℃－２週間及び４０℃－４週間保存）を行った。そして、熱加速前後における組換えモノクローナル抗体の純度を、サイズ排除クロマトグラフ法（ＳＥＣ）及びイオン交換クロマトグラフ法（ＩＥＸ）で確認した。また、液体製剤の使用性を動粘度により評価した。サイズ排除クロマトグラフ法（ＳＥＣ）、イオン交換クロマトグラフ法（ＩＥＸ）及び動粘度の分析条件は以下の通りである。 In order to evaluate the stability of the liquid preparation, 0.5 mL of each evaluation sample is filled into a 2 mL glass vial, and thermal acceleration tests (60 ° C.-2 weeks, 50 ° C.-2 weeks, and 40 ° C.-4) Weekly). The purity of the recombinant monoclonal antibody before and after thermal acceleration was confirmed by size exclusion chromatography (SEC) and ion exchange chromatography (IEX). Moreover, the usability of the liquid preparation was evaluated by kinematic viscosity. The analysis conditions for size exclusion chromatography (SEC), ion exchange chromatography (IEX) and kinematic viscosity are as follows.
［サイズ排除クロマトグラフ法］
　評価試料を移動相でタンパク質濃度が１ｍｇ／ｍＬとなるように希釈し評価試料溶液とした。
　評価試料溶液２０μＬにつき、以下の条件で液体クロマトグラフ法により試験を行い、高分子画分、メイン画分、低分子画分のピーク面積を自動分析法により測定し、その量（％）を求めた。 [Size Exclusion Chromatography]
The evaluation sample was diluted with a mobile phase so that the protein concentration became 1 mg / mL, and used as an evaluation sample solution.
Test 20 μL of the evaluation sample solution by the liquid chromatograph method under the following conditions, measure the peak area of the high molecular fraction, the main fraction, and the low molecular fraction by the automatic analysis method, and obtain the amount (%). It was.
分析条件
カラム：ＴＳＫｇｅｌ　Ｇ３０００ＳＷｘｌ　７．８ｍｍ　Ｉ．Ｄ．　×　３０　ｃｍ　（東ソー製）
ガードカラム：ＴＳＫｇｅｌ　ｇｕａｒｄ　ｃｏｌｕｍｎ　ＳＷＸＬ　６．０ｍｍ　Ｉ．Ｄ．　×　４ｃｍ（東ソー製）
移動相：ｐＨ６．８のリン酸緩衝液（３００ｍｍｏｌ／Ｌ塩化ナトリウムを含むｐＨ６．８の２０ｍｍｏｌ／Ｌリン酸緩衝液）
評価試料注入量：組換えモノクローナル抗体にして約２０μｇ
流量：０．５ｍＬ／ｍｉｎ
検出波長：２８０ｎｍ Analysis condition column: TSKgel G3000SWxl 7.8 mm D. × 30 cm (Tosoh product)
Guard column: TSKgel guard column SWXL 6.0 mm D. × 4cm (Made by Tosoh)
Mobile phase: pH 6.8 phosphate buffer (pH 6.8, 20 mmol / L phosphate buffer with 300 mmol / L sodium chloride)
Evaluation sample injection amount: about 20 μg as a recombinant monoclonal antibody
Flow rate: 0.5mL / min
Detection wavelength: 280 nm
計算式
各ピークの合計面積＝メイン画分のピーク面積＋高分子画分のピーク面積＋低分子画分のピーク面積
高分子画分（％）＝（高分子画分の各ピーク面積の合計／各ピークの合計面積）×１００
低分子画分（％）＝（低分子画分の各ピーク面積の合計／各ピークの合計面積）×１００ Calculation formula Total area of each peak = peak area of main fraction + peak area of high molecular fraction + peak area of low molecular fraction high molecular fraction (%) = (total of peak areas of high molecular fraction / Total area of each peak) x 100
Low molecular fraction (%) = (total of each peak area of low molecular fraction / total area of each peak) × 100
［動粘度測定法］
　評価試料をそのまま評価試料溶液とした。
　ガラスシリンジに注射針を装着した状態で評価試料溶液を吸引する際に押し子に発生する張力を測定した。得られた張力を、動粘度２ｍｍ２／ｓ～２０ｍｍ２／ｓの粘度計校正用標準溶液（日本グリース）を同様に測定して得た張力と動粘度とから作成した検量線に当てはめ、動粘度を求めた。動粘度の測定温度は、２０℃とした。 [Kinematic viscosity measurement method]
The evaluation sample was directly used as an evaluation sample solution.
The tension generated in the pusher when the evaluation sample solution was sucked with the injection needle attached to the glass syringe was measured. The obtained tension was applied to a calibration curve prepared from the tension and kinematic viscosity obtained by measuring a viscometer calibration standard solution (Nippon Grease) having a kinematic viscosity of 2 mm 2 / s to 20 mm 2 / s in the same manner. The viscosity was determined. The kinematic viscosity measurement temperature was 20 ° C.
　本実施例で得られたサイズ排除クロマトグラフ法の評価結果を表２に、イオン交換クロマトグラフ法の評価結果を表３に、動粘度の評価結果を表４に示した。 The evaluation results of the size exclusion chromatography method obtained in this example are shown in Table 2, the evaluation results of the ion exchange chromatography method are shown in Table 3, and the kinematic viscosity evaluation results are shown in Table 4.
Figure JPOXMLDOC01-appb-T000002
Figure JPOXMLDOC01-appb-T000002
Figure JPOXMLDOC01-appb-T000003
Figure JPOXMLDOC01-appb-T000003
Figure JPOXMLDOC01-appb-T000004
Figure JPOXMLDOC01-appb-T000004
　９４ｍＭのアミノ酸成分を含有し、ｐＨを６に調整した処方（評価試料Ｎｏ．５）は、アミノ酸量の合計量もヒスチジン成分の含有量も少ないにもかかわらず、６０℃で２週間、５０℃で２週間及び４０℃で４週間保存した熱加速試験において、高分子画分及び低分子画分の量が、アミノ酸の合計量もヒスチジン成分の含有量も多く使っている評価試料Ｎｏ．１に比べて同程度であった。
　更に、評価試料Ｎｏ．５は、イオン交換クロマトグラフ法における脱アミド体を含む酸性画分の増加が、評価試料Ｎｏ．１と同様に抑制されていることが確認された。
　また、ｐＨを５に調整した処方（評価試料Ｎｏ．２）及びｐＨを６．８に調製した処方（評価試料Ｎｏ，３及び４）では、二量体を含む高分子画分の評価試料Ｎｏ．５に比べると増加傾向を認め、脱アミド体を含む酸性画分にも増加傾向を認めたため、ｐＨの範囲は重要な管理項目であることが明らかになった。
　ヒスチジンの添加の有無を比較した処方（評価試料Ｎｏ．３及び４）の評価結果から、ヒスチジン成分は期待するｐＨ範囲に液体製剤のｐＨを調整する点から、重要な成分であることが明らかになった。
　また、評価試料Ｎｏ．５の動粘度は、評価試料Ｎｏ．１の動粘度に比べて低く、その変化も小さいので、皮下注射を行うときの使用性が向上していた。
　このように、アミノ酸成分の含有量を９４ｍＭ以下にしても、ｐＨを５．５～６．７の範囲内に調整することにより皮下注射可能な動粘度を示す液体製剤を調製し得ることが明らかになった。 The formulation containing 94 mM amino acid component and adjusted to pH 6 (evaluation sample No. 5) had a total amount of amino acids and a small content of histidine component, but at 60 ° C. for 2 weeks, 50 ° C. In the thermal acceleration test stored for 2 weeks at 40 ° C. and 4 weeks at 40 ° C., the evaluation samples No. 1 and No. 2 in which the amount of the high molecular fraction and the low molecular fraction are large in both the total amount of amino acids and the content of the histidine component are used. It was comparable to 1.
Furthermore, the evaluation sample No. 5 shows that the increase in the acidic fraction containing the deamidated product in the ion-exchange chromatography method was evaluated sample No. It was confirmed that it was suppressed similarly to 1.
Moreover, in the prescription (evaluation sample No. 2) adjusted to pH 5 and the prescription (evaluation sample No. 3, 3 and 4) adjusted to pH 6.8, evaluation sample No. of the polymer fraction containing the dimer . Since an increasing tendency was observed as compared to 5, and an increasing tendency was also observed in the acidic fraction containing the deamidated substance, it was revealed that the pH range was an important management item.
From the evaluation results of the formulation (evaluation sample No. 3 and 4) comparing the presence or absence of histidine addition, it is clear that the histidine component is an important component from the point of adjusting the pH of the liquid preparation to the expected pH range became.
In addition, evaluation sample No. The kinematic viscosity of No. Since it was lower than the kinematic viscosity of 1 and its change was small, the usability when subcutaneous injection was performed was improved.
Thus, it is clear that even when the content of the amino acid component is 94 mM or less, a liquid preparation showing kinematic viscosity that can be injected subcutaneously can be prepared by adjusting the pH within the range of 5.5 to 6.7. Became.
（実施例２）
アミノ酸成分の含有量をより低減した場合の安定性効果の確認
　アミノ酸成分の含有量を９４ｍＭよりさらに低減させた場合の組換えモノクローナル抗体（トシリズマブとして１８０ｍｇ／ｍＬ）を含む液体製剤について、安定化効果を評価した。
　本検討では、アミノ酸成分濃度をより低減させる可能性を評価するため、評価試料Ｎｏ．６－１～Ｎｏ．８を調製した。各評価試料の処方は以下の通りである。 (Example 2)
Confirmation of stability effect when content of amino acid component is further reduced Stabilizing effect for liquid preparation containing recombinant monoclonal antibody (180 mg / mL as tocilizumab) when content of amino acid component is further reduced below 94 mM Evaluated.
In this study, in order to evaluate the possibility of further reducing the amino acid component concentration, evaluation sample No. 6-1 to No. 8 was prepared. The prescription of each evaluation sample is as follows.
Figure JPOXMLDOC01-appb-T000005
Figure JPOXMLDOC01-appb-T000005
　液体製剤の安定性を評価するために、各評価試料を２ｍＬガラスバイアルに０．５ｍＬ充填して、各評価試料の熱加速試験（６０℃－１週間、５０℃－２週間、４０℃－８週間及び２５℃－４ヶ月保存）を行った。熱加速前後における組換えモノクローナル抗体の純度を、サイズ排除クロマトグラフ法及びイオン交換クロマトグラフ法により評価し、また、その使用性を動粘度により評価した。分析条件は、実施例１に示した通りである。 In order to evaluate the stability of the liquid preparation, 0.5 mL of each evaluation sample is filled into a 2 mL glass vial, and a thermal acceleration test (60 ° C.-1 week, 50 ° C.-2 weeks, 40 ° C.-8) Weekly and at 25 ° C. for 4 months). The purity of the recombinant monoclonal antibody before and after thermal acceleration was evaluated by size exclusion chromatography and ion exchange chromatography, and its usability was evaluated by kinematic viscosity. The analysis conditions are as shown in Example 1.
　本実施例で得られたサイズ排除クロマトグラフ法の評価結果を表６に、イオン交換クロマトグラフ法の評価結果を表７に、動粘度の評価結果を表８に示した。 The evaluation results of the size exclusion chromatography method obtained in this example are shown in Table 6, the evaluation results of the ion exchange chromatography method are shown in Table 7, and the kinematic viscosity evaluation results are shown in Table 8.
Figure JPOXMLDOC01-appb-T000006
Figure JPOXMLDOC01-appb-T000006
Figure JPOXMLDOC01-appb-T000007
Figure JPOXMLDOC01-appb-T000007
Figure JPOXMLDOC01-appb-T000008
Figure JPOXMLDOC01-appb-T000008
　いずれの処方も、表６に示すように、実施例１に示したアミノ酸成分濃度が９４ｍＭよりアミノ酸含量が多い評価試料（評価試料Ｎｏ．１）と比較すると、サイズ排除クロマトグラフ法の評価において、熱安定性が同一条件の試験である５０℃－２週間の評価から見て高分子画分の増加が抑制されことが確認された。
　この結果より、アミノ酸成分の含有量を９３ｍＭ以下にすることにより、液体製剤中の二量体生成が抑制され得ることが明らかになった。
　また、表７に示すように、評価試料Ｎｏ．１と比較し、脱アミド体が含まれる酸性画分の生成抑制効果が熱安定性が同一条件の試験である５０℃－２週間の評価から見てさらに改善されることが確認できた。なお、動粘度については、いずれの評価試料も著しい増加を認めておらず、皮下注射を行う上で問題ない使用性を示すことが確認された。
　アミノ酸成分濃度を９０ｍＭから更に低減させた処方（評価試料Ｎｏ．７及び８）の評価結果から、アミノ酸成分濃度を９０ｍＭから更に下げることで、加速条件において、サイズ排除クロマトグラフ法の評価における二量体を含む高分子画分の増加傾向を認めたが、スクロース（精製白糖）を加えることで、増加効果を抑制できることが確認された。
　さらに、５５．６６ｍｇ／ｍＬのスクロースを加えても、動粘度に著しい増加を認めておらず、皮下注射を行う上で、問題のない使用性を示すことが確認された。アミノ酸成分とスクロースとを組み合わせることにより、二量体生成を抑制し得る効果を奏し得ることが明らかになった。 As shown in Table 6, each formulation was compared with the evaluation sample (evaluation sample No. 1) in which the amino acid component concentration shown in Example 1 had an amino acid content higher than 94 mM. In the evaluation of the size exclusion chromatography method, From the evaluation at 50 ° C. for 2 weeks, which is a test under the same thermal stability conditions, it was confirmed that the increase in the polymer fraction was suppressed.
From this result, it was revealed that dimer formation in the liquid preparation can be suppressed by setting the content of the amino acid component to 93 mM or less.
As shown in Table 7, the evaluation sample No. Compared to 1, it was confirmed that the effect of suppressing the formation of the acidic fraction containing the deamidated compound was further improved in view of evaluation at 50 ° C. for 2 weeks, which is a test under the same thermal stability. In addition, regarding the kinematic viscosity, no significant increase was observed in any of the evaluation samples, and it was confirmed that there was no problem in using the subcutaneous injection.
From the evaluation results of the formulations (evaluation sample Nos. 7 and 8) in which the amino acid component concentration is further reduced from 90 mM, the dimer in the evaluation of the size exclusion chromatographic method under accelerated conditions by further reducing the amino acid component concentration from 90 mM. Although the increase tendency of the polymer fraction containing a body was recognized, it was confirmed that the increase effect can be suppressed by adding sucrose (purified sucrose).
Furthermore, even when 55.66 mg / mL sucrose was added, no significant increase in kinematic viscosity was observed, confirming that there was no problem in use for subcutaneous injection. It has been clarified that by combining the amino acid component and sucrose, an effect of suppressing dimer formation can be obtained.
　本発明の製剤は、長期保存時の二量体生成抑制及び脱アミド化抑制を実現した安定な高濃度組換えモノクローナル抗体液体製剤であり、且つ、皮下注射を可能にする動粘度を示す高濃度組換えモノクローナル抗体含有液体製剤を低コストで提供できるので、産業上の有用性が高い。 The preparation of the present invention is a stable high-concentration recombinant monoclonal antibody liquid preparation that realizes suppression of dimer formation and deamidation during long-term storage, and also exhibits high kinematic viscosity that enables subcutaneous injection Since a liquid preparation containing a recombinant monoclonal antibody can be provided at low cost, it is highly industrially useful.

Claims (11)

  1. 　１５０ｍｇ／ｍＬ～２００ｍｇ／ｍＬの組換えモノクローナル抗体と、ヒスチジン成分が５ｍＭ未満である４５ｍＭ～９４ｍＭのアミノ酸成分とを含有し、ｐＨが５．５～６．７である高濃度組換えモノクローナル抗体含有液体製剤。 Contains a recombinant monoclonal antibody of 150 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 a high concentration recombinant monoclonal antibody having a pH of 5.5 to 6.7 Liquid formulation.
  2. 　ヒスチジン成分以外のアミノ酸成分が、アルギニン、アルギニン塩酸塩及びメチオニンからなる群より選ばれる少なくとも１種である請求項１に記載の液体製剤。 The liquid preparation according to claim 1, 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.
  3. 　ヒスチジン成分が、ヒスチジン及びヒスチジン塩酸塩からなる群より選ばれる少なくとも１種である請求項１又は請求項２に記載の液体製剤。 The liquid preparation according to claim 1 or 2, wherein the histidine component is at least one selected from the group consisting of histidine and histidine hydrochloride.
  4. 　７５ｍＭ～９３ｍＭのアミノ酸成分を含有する請求項１ないし請求項３のいずれか１項に記載の液体製剤。 The liquid preparation according to any one of claims 1 to 3, comprising an amino acid component of 75 mM to 93 mM.
  5. 　９０ｍＭのアミノ酸成分を含有する請求項１ないし請求項４のいずれか１項に記載の液体製剤。 The liquid preparation according to any one of claims 1 to 4, comprising a 90 mM amino acid component.
  6. 　ｐＨが５．８～６．４である請求項１ないし請求項５のいずれか１項に記載の液体製剤。 The liquid preparation according to any one of claims 1 to 5, which has a pH of 5.8 to 6.4.
  7. 　ｐＨが６．０～６．２である請求項１ないし請求項６のいずれか１項に記載の液体製剤。 The liquid preparation according to any one of claims 1 to 6, having a pH of 6.0 to 6.2.
  8. 　１７０ｍｇ／ｍＬ～１９０ｍｇ／ｍＬの組換えモノクローナル抗体を含有する請求項１ないし請求項７のいずれか１項に記載の液体製剤。 The liquid preparation according to any one of claims 1 to 7, which contains 170 to 190 mg / mL recombinant monoclonal antibody.
  9. 　１８０ｍｇ／ｍＬの組換えモノクローナル抗体を含有する請求項１ないし請求項８のいずれか１項に記載の液体製剤。 The liquid preparation according to any one of claims 1 to 8, comprising 180 mg / mL recombinant monoclonal antibody.
  10. 　さらにポリオールを含有する請求項１ないし請求項９のいずれか１項に記載の液体製剤。 The liquid preparation according to any one of claims 1 to 9, further comprising a polyol.
  11. 　組換えモノクローナル抗体が、トシリズマブ、トラスツズマブ、リツキシマブ、パリビズマブ、インフリキシマブ、バシリキシマブ、ゲムツズマブオゾガマイシン、ベバシズマブ、イブリツモマブ　チウキセタン、アダリムマブ、セツキシマブ、ラニビズマブ、オマリズマブ、エクリズマブ、パニツムマブ、ウステキヌマブ、ゴリムマブ、カナキヌマブ、デノスマブ、モガムリズマブ、オファツムマブ、ペルツズマブ、トラスツズマブ　エムタンシン、ブレンツキシマブ　ベドチン、ナタリズマブ、ニボルマブ、アレムツズマブ、セクキヌマブ、ラムシルマブ及びイピリムマブからなる群から選択される少なくとも一つである請求項１０に記載の液体製剤。 Recombinant monoclonal antibodies are tocilizumab, trastuzumab, rituximab, rituximab, rituximab, paclizumab, infliximab, basiliximab, gemtuzumab ozogamicin, bevacizumab, ibritumomab, rituximab, rituximab, raribizumab The liquid preparation according to claim 10, which is at least one selected from the group consisting of: mogamulizumab, ofatumumab, pertuzumab, trastuzumab emtansine, brentuximab vedotin, natalizumab, nivolumab, alemtuzumab, secukinumab, ramcilmab, and ipilimumab.
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