Stable liquid pharmaceutical preparations

Abstract

The stable liquid pharmaceutical formulation according to the present invention comprises: an antibody or an antigen-binding fragment thereof; a surfactant; a sugar or a derivative thereof; and a buffer. The stable liquid pharmaceutical formulation according to the present invention has low viscosity, contains a high amount of antibody, exhibits excellent long-term storage stability based on its excellent stability under accelerated and extreme conditions, and can be administered subcutaneously.

Description

Technical Field

[0001] This invention relates to stable liquid pharmaceutical formulations. Background Technology

[0002] Tumor necrosis factor-α (TNF-α) is a cell signaling protein (cytokine) involved in systemic inflammation and a cytokine mediating acute-phase responses. TNF-α is associated with various diseases and conditions, including sepsis, infections, autoimmune diseases, and transplant rejection. TNF-α stimulates immune responses and causes many clinical problems associated with autoimmune abnormalities, such as rheumatoid arthritis, ankylosing spondylitis, ulcerative colitis, adult Crohn's disease, pediatric Crohn's disease, psoriasis, and psoriatic arthritis. TNF-α inhibitors can be used to treat these abnormalities.

[0003] Infliximab is a chimeric monoclonal antibody that can function as a TNF-α inhibitor. A standard formulation containing this antibody is prepared as a lyophilized powder, reconstituted, diluted, and administered intravenously using a dosage regimen determined for each disease.

[0004] For example, the Remicade trademark discloses a lyophilized formulation containing infliximab, sucrose, polysorbate 80, and sodium phosphate. For intravenous injection, it discloses a reconstitution step of adding injectable water to the lyophilized formulation, and a step of diluting the reconstituted formulation with injectable saline containing sodium chloride.

[0005] However, the conventional administration method of the preparation described above (freeze-drying → reconstitution → dilution → intravenous administration) is problematic because it is expensive, complex, and causes patient discomfort, rejection, and side effects due to frequent administration. Furthermore, the administration of this preparation is limited to medically trained individuals.

[0006] Adalimumab is also a human monoclonal antibody that can act as a TNF-α inhibitor. Liquid formulations containing adalimumab are disclosed, for example, in the Humira trademark. Furthermore, Korean Patent Application Publication No. 10-2014-0134689 discloses a liquid formulation containing adalimumab, sodium phosphate, sodium citrate, citric acid, mannitol, sodium chloride, and polysorbate 80 (Example 1), and an improved liquid formulation containing adalimumab, sodium phosphate, sodium citrate, citric acid, mannitol, arginine, sodium chloride, and polysorbate 80 (Example 2).

[0007] However, in the case of liquid drug formulations containing NaCl or KCl as isotonic agents, problems such as precipitation and gelation may occur, and when the antibody concentration is low to about 50 mg / ml, the frequency and cycle of administration may be limited.

[0008] Therefore, there is a need for a stable liquid drug formulation that overcomes the problems of conventional liquid drug formulations and contains an antibody as a TNF-α inhibitor, particularly infliximab. Summary of the Invention

[0009] Technical issues

[0010] One object of the present invention is to provide a stable liquid pharmaceutical formulation having low viscosity and containing a high content of antibodies.

[0011] Another object of the present invention is to provide a liquid pharmaceutical formulation that exhibits excellent long-term storage stability based on its superior stability under accelerated and harsh conditions.

[0012] Another object of the present invention is to provide a stable liquid pharmaceutical formulation that can be administered subcutaneously.

[0013] Technical solution

[0014] According to one embodiment of the present invention, a stable liquid pharmaceutical formulation comprises: (A) an antibody or an antigen-binding fragment thereof; (B) a surfactant; (C) a sugar or a derivative thereof; and (D) a buffer.

[0015] In one embodiment of the invention, antibody (A) may comprise an antibody that binds to TNF-α.

[0016] In one embodiment of the present invention, antibody (A) may comprise infliximab, adalimumab, PEGylated cetrus Fab fragment, golimumab, or a mixture thereof.

[0017] In one embodiment of the invention, antibody (A) may comprise a chimeric human-mouse IgG monoclonal antibody.

[0018] In one embodiment of the present invention, the antibody or its antigen-binding fragment (A) may comprise: a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises a CDR1 domain containing the amino acid sequence shown in SEQ ID NO: 1, a CDR2 domain containing the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 domain containing the amino acid sequence shown in SEQ ID NO: 3; and the heavy chain variable region comprises a CDR1 domain containing the amino acid sequence shown in SEQ ID NO: 4, a CDR2 domain containing the amino acid sequence shown in SEQ ID NO: 5, and a CDR3 domain containing the amino acid sequence shown in SEQ ID NO: 6.

[0019] In one embodiment of the present invention, the antibody or its antigen-binding fragment (A) may comprise: a light chain variable region having the amino acid sequence shown in SEQ ID NO: 7; and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 8.

[0020] In one embodiment of the present invention, the antibody (A) may comprise: a light chain having the amino acid sequence shown in SEQ ID NO: 9; and a heavy chain having the amino acid sequence shown in SEQ ID NO: 10.

[0021] In one embodiment of the invention, the concentration of the antibody or its antigen-binding fragment (A) may be 10 to 200 mg / ml.

[0022] In one embodiment of the invention, surfactant (B) may comprise polysorbate, poloxamer, or a mixture thereof.

[0023] In one embodiment of the invention, the surfactant (B) may comprise polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, or a mixture of two or more thereof.

[0024] In one embodiment of the invention, surfactant (B) may comprise polysorbate 80.

[0025] In one embodiment of the invention, the concentration of the surfactant (B) may be 0.02 to 0.1% (w / v).

[0026] In one embodiment of the present invention, sugar (C) may comprise monosaccharides, disaccharides, oligosaccharides, polysaccharides, or mixtures of two or more thereof, and sugar derivatives (C) may comprise sugar alcohols, sugar acids, or mixtures thereof.

[0027] In one embodiment of the invention, the sugar or its derivative (C) may comprise sorbitol, mannitol, trehalose, sucrose, or a mixture of two or more thereof.

[0028] In one embodiment of the invention, the concentration of the sugar or its derivative (C) may be 1 to 10% (w / v).

[0029] In one embodiment of the invention, the buffer (D) may comprise acetate or histidine.

[0030] In one embodiment of the invention, the buffer (D) may have a concentration of 1 to 50 mM.

[0031] In one embodiment of the invention, the formulation may have a pH of 4.0 to 5.5.

[0032] In one embodiment of the invention, the formulation may be free of aspartic acid, lysine, arginine, or mixtures thereof.

[0033] In one embodiment of the present invention, the formulation may be free of NaCl, KCl, NaF, KBr, NaBr, Na2SO4, NaSCN, K2SO4 or mixtures thereof.

[0034] In one embodiment of the invention, the formulation may be free of chelating agents.

[0035] In one embodiment of the invention, the formulation has a viscosity of 0.5 cp to 10 cp after being stored at 40°C ± 2°C for 1 month, or a viscosity of 0.5 cp to 5 cp after being stored at 5°C ± 3°C for 6 months.

[0036] According to one embodiment of the present invention, a stable liquid pharmaceutical formulation may comprise: (A) an antibody or an antigen-binding fragment thereof, comprising a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises a CDR1 domain containing the amino acid sequence shown in SEQ ID NO: 1, a CDR2 domain containing the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 domain containing the amino acid sequence shown in SEQ ID NO: 3; the heavy chain variable region comprises a CDR1 domain containing the amino acid sequence shown in SEQ ID NO: 4, a CDR2 domain containing the amino acid sequence shown in SEQ ID NO: 5, and a CDR3 domain containing the amino acid sequence shown in SEQ ID NO: 6; (B) a surfactant; (C) a sugar or a derivative thereof; and (D) a buffer comprising acetate or histidine.

[0037] According to one embodiment of the present invention, a stable liquid pharmaceutical formulation may comprise: (A) 90 to 145 mg / ml of an antibody or an antigen-binding fragment thereof, comprising a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises a CDR1 domain containing the amino acid sequence shown in SEQ ID NO: 1, a CDR2 domain containing the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 domain containing the amino acid sequence shown in SEQ ID NO: 3; the heavy chain variable region comprises a CDR1 domain containing the amino acid sequence shown in SEQ ID NO: 4, a CDR2 domain containing the amino acid sequence shown in SEQ ID NO: 5, and a CDR3 domain containing the amino acid sequence shown in SEQ ID NO: 6; (B) 0.02 to 0.1% (w / v) of a surfactant; (C) 1 to 10% (w / v) of a sugar or a derivative thereof; and (D) 1 to 50 mM of a buffer comprising acetate or histidine.

[0038] In one embodiment of the invention, the stable liquid pharmaceutical formulation can be used for subcutaneous administration.

[0039] In one embodiment of the invention, the stable liquid pharmaceutical preparation may not undergo a reconstitution step, a dilution step, or both before use.

[0040] According to one embodiment of the present invention, a pre-filled syringe is filled with the aforementioned stable liquid pharmaceutical formulation.

[0041] An auto-injector according to one embodiment of the present invention includes the pre-filled syringe.

[0042] Beneficial effects

[0043] The stable liquid pharmaceutical formulation according to the present invention has low viscosity and contains a high content of antibodies. Based on its excellent stability under accelerated and harsh conditions, it has excellent long-term storage stability and can be administered subcutaneously. Detailed Implementation

[0044] [Stable liquid pharmaceutical preparations]

[0045] The stable liquid pharmaceutical formulation according to the present invention contains: (A) an antibody or an antigen-binding fragment thereof; (B) a surfactant; (C) a sugar or a derivative thereof; and (D) a buffer.

[0046] As used herein, the term "free of" means that the formulation contains no corresponding component at all. Furthermore, the term means that the formulation substantially contains no corresponding component, i.e., the amount of the corresponding component present does not affect antibody activity or the stability and viscosity of the liquid drug formulation. For example, the term means that the formulation contains the corresponding component in an amount of 0 to 1% (w / v), 0 to 1 ppm (w / v), or 0 to 1 ppb (w / v) based on the total weight of the liquid drug formulation.

[0047] (A) Antibody or its antigen-binding fragment

[0048] The term "antibody" refers to an immunoglobulin molecule composed of four polypeptide chains, two heavy chains and two light chains linked together by disulfide bonds. Other naturally occurring antibodies with altered structures, such as camel antibodies, are also included in this definition. Each heavy chain consists of a heavy chain variable region and a heavy chain constant region. The heavy chain constant region consists of three domains (CH1, CH2, and CH3). Each light chain consists of a light chain variable region and a light chain constant region. The light chain constant region consists of one domain (CL). The heavy chain variable region and light chain variable region can be further subdivided into hypervariable regions called complementarity-determining regions (CDRs), interspersed with more conserved regions called framework regions (FRs). Each heavy chain variable region and light chain variable region consists of three CDRs and four FRs, arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[0049] In one embodiment of the invention, the pharmaceutical formulation may contain polyclonal antibodies, monoclonal antibodies, recombinant antibodies, single-chain antibodies, hybrid antibodies, chimeric antibodies, humanized antibodies, or fragments thereof as antibodies. The term "chimeric antibody" refers to an antibody comprising variable region sequences of heavy and light chains from one species and constant region sequences from another species. In one embodiment of the invention, the pharmaceutical formulation may contain a chimeric human-mouse IgG monoclonal antibody as an antibody. The chimeric human-mouse IgG monoclonal antibody consists of mouse heavy and light chain variable regions and human heavy and light chain constant regions bound thereto. Chimeric human-mouse IgG monoclonal antibodies can be produced according to methods known in the art. For example, infliximab can be produced according to the method described in U.S. Patent No. 6,284,471.

[0050] In one embodiment of the invention, the pharmaceutical formulation may contain an antibody that binds to TNF-α or a TNF-α epitope. The antibody binding to TNF-α or a TNF-α epitope may comprise infliximab, adalimumab, PEGylated sertozumab Fab fragment, golimumab, or a mixture thereof. In one embodiment of the invention, the antibody may comprise infliximab.

[0051] In one embodiment of the present invention, the antibody or its antigen-binding fragment (A) may comprise: a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises a CDR1 domain containing the amino acid sequence shown in SEQ ID NO: 1, a CDR2 domain containing the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 domain containing the amino acid sequence shown in SEQ ID NO: 3; and the heavy chain variable region comprises a CDR1 domain containing the amino acid sequence shown in SEQ ID NO: 4, a CDR2 domain containing the amino acid sequence shown in SEQ ID NO: 5, and a CDR3 domain containing the amino acid sequence shown in SEQ ID NO: 6.

[0052] In one embodiment of the present invention, the antibody or its antigen-binding fragment (A) may comprise: a light chain variable region having the amino acid sequence shown in SEQ ID NO: 7; and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 8.

[0053] In one embodiment of the present invention, the antibody or its antigen-binding fragment (A) may comprise: a light chain having the amino acid sequence shown in SEQ ID NO: 9; and a heavy chain having the amino acid sequence shown in SEQ ID NO: 10.

[0054] The concentration of the antibody or its antigen-binding fragment can be freely controlled within a range that does not substantially adversely affect the stability and viscosity of the stable liquid pharmaceutical formulation according to the invention. In one embodiment of the invention, the concentration of the antibody or its antigen-binding fragment can be 10 to 200 mg / ml. In another embodiment of the invention, the concentration of the antibody or its antigen-binding fragment can be 50 to 200 mg / ml. In another embodiment of the invention, the concentration of the antibody or its antigen-binding fragment can be 80 to 150 mg / ml. In another embodiment of the invention, the concentration of the antibody or its antigen-binding fragment can be 90 to 145 mg / ml. In another embodiment of the invention, the concentration of the antibody or its antigen-binding fragment can be 110 to 130 mg / ml. If the concentration of the antibody or its antigen-binding fragment is within the above ranges, the high content of the antibody or its antigen-binding fragment allows for increased freedom in dosage and administration cycle, and the pharmaceutical formulation can exhibit excellent long-term stability and low viscosity.

[0055] (B) Surfactants

[0056] Examples of surfactants include, but are not limited to, polyoxyethylene dehydrated sorbitol fatty acid esters (e.g., polysorbate esters), polyoxyethylene alkyl ethers (e.g., Brij), alkylphenyl polyoxyethylene ethers (e.g., Triton-X), polyoxyethylene-polyoxypropylene copolymers (e.g., Poloxamer, Pluronic), sodium dodecyl sulfate (SDS), etc.

[0057] In one embodiment of the invention, the surfactant may comprise polyoxyethylene dehydrated sorbitol fatty acid ester (polysorbate). The polysorbate may comprise polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, or a mixture of two or more thereof. In one embodiment of the invention, the polysorbate may comprise polysorbate 20, polysorbate 80, or a mixture thereof. In another embodiment of the invention, the polysorbate may comprise polysorbate 80.

[0058] In one embodiment of the invention, the concentration of the surfactant can be freely controlled within a range that does not substantially adversely affect the stability and viscosity of the stable liquid pharmaceutical formulation according to the invention. For example, the concentration of the surfactant can be 0.001 to 5% (w / v), 0.01 to 1% (w / v), or 0.02 to 0.1% (w / v). If the concentration of the surfactant is within the above range, the pharmaceutical composition can exhibit excellent long-term stability and low viscosity.

[0059] (C) Sugars or their derivatives

[0060] Sugars may include monosaccharides, disaccharides, oligosaccharides, polysaccharides, or mixtures of two or more thereof. Examples of monosaccharides include, but are not limited to, glucose, fructose, and galactose. Examples of disaccharides include, but are not limited to, sucrose, lactose, maltose, and trehalose. Examples of oligosaccharides include, but are not limited to, fructooligosaccharides, galactooligosaccharides, and mannose. Examples of polysaccharides include, but are not limited to, starch, glycogen, cellulose, chitin, and pectin.

[0061] Sugar derivatives may include sugar alcohols, sugar acids, or mixtures thereof. Examples of sugar alcohols include, but are not limited to, glycerol, erythritol, threitol, arabinitol, xylitol, ribitol, mannitol, sorbitol, galactitol, fucitol, idutol, inositol, heptaheptaol, isomaltulitol, maltitol, lactitol, maltotriol, maltotetratitol, polydextrose, etc. Examples of sugar acids include, but are not limited to, aldonic acids (glyceric acid, etc.), ketonic acids (neuraminic acid, etc.), uronic acids (glucuronic acid, etc.), aldonic diacids (tartaric acid, etc.), etc.

[0062] In one embodiment of the invention, the sugar or its derivative (C) may comprise sorbitol, mannitol, trehalose, sucrose, or a mixture of two or more thereof.

[0063] In one embodiment of the invention, the concentration of sugar or its derivatives can be freely controlled within a range that does not substantially adversely affect the stability and viscosity of the stable liquid pharmaceutical formulation according to the invention. For example, the concentration of sugar or its derivatives can be 0.1 to 30% (w / v), 1 to 20% (w / v), or 1 to 10% (w / v). If the concentration of sugar or its derivatives is within this range, the pharmaceutical composition can exhibit excellent long-term stability and low viscosity.

[0064] (D) Buffer

[0065] The buffers used in this invention are neutralizing substances that minimize pH changes caused by acids or bases. Examples of buffers include phosphates, acetates, succinates, gluconates, glutamates, citrates, histidines, etc. In one embodiment of the invention, the buffer may contain acetate or histidine. If the buffer contains both acetate and histidine, the stability of the pharmaceutical formulation is reduced.

[0066] In one embodiment of the invention, the buffer may comprise an acetate. Examples of acetates include, but are not limited to, sodium acetate, zinc acetate, aluminum acetate, ammonium acetate, potassium acetate, etc. For pH adjustment, the buffer may further comprise an acid, such as acetic acid. When the buffer comprises an acetate, it is most preferred in terms of pH adjustment and stability.

[0067] In one embodiment of the invention, the buffer may contain histidine. When the buffer contains histidine, it may contain histidine salts, such as histidine chloride, histidine acetate, histidine phosphate, histidine sulfate, etc. For pH adjustment, the buffer may contain acids, such as hydrochloric acid, acetic acid, phosphoric acid, sulfuric acid, etc.

[0068] In one embodiment of the invention, the stable liquid pharmaceutical formulation may be free of citrate, phosphate, or mixtures thereof.

[0069] In one embodiment of the invention, the concentration of the buffer (or the anion of the buffer) can be freely controlled within a range that does not substantially adversely affect the stability and viscosity of the stable liquid pharmaceutical formulation according to the invention. For example, the concentration of the buffer or its anion can be 1 to 50 mM, 5 to 30 mM, or 10 to 25 mM. If the concentration of the buffer or its anion is within this range, the pharmaceutical composition can exhibit excellent long-term stability and low viscosity.

[0070] (E)pH

[0071] In one embodiment of the invention, the pH of the stable liquid pharmaceutical composition can be 4.0 to 5.5, or 4.7 to 5.3. If the pH is within this range, the pharmaceutical composition exhibits excellent long-term stability and low viscosity. The pH of the pharmaceutical formulation can be adjusted using a buffer. In other words, if the pharmaceutical formulation contains a certain amount of buffer, it can exhibit a pH within the aforementioned range without the need for a separate pH adjuster. If citrate, phosphate, or mixtures thereof are used as buffers, it may be difficult to achieve a pH within the aforementioned range. If the pharmaceutical formulation also contains an acid (e.g., hydrochloric acid) or a base (e.g., sodium hydroxide) as a separate pH adjuster, the stability of the antibody is reduced.

[0072] (F) Other components

[0073] In one embodiment of the invention, the stable liquid pharmaceutical formulation may be free of aspartic acid, lysine, arginine, or mixtures thereof. If the stable liquid pharmaceutical formulation contains these amino acids, it may become a solid. In another embodiment of the invention, the stable liquid pharmaceutical formulation may contain one or more amino acids other than the three amino acids mentioned above. In this case, the stable liquid pharmaceutical formulation may contain one or more amino acids in an amount of 5% (w / v) or less, for example, 0.001 to 5% (w / v), 0.001 to 1% (w / v), 0.01 to 5% (w / v), 0.01 to 1% (w / v), 0.1 to 5% (w / v), or 0.1 to 1% (w / v).

[0074] In another embodiment of the invention, the stable liquid pharmaceutical formulation may contain taurine. In this case, the amount of taurine may be 5% (w / v) or lower, for example, 0.001 to 5% (w / v), 0.001 to 1% (w / v), 0.01 to 5% (w / v), 0.01 to 1% (w / v), 0.1 to 5% (w / v), or 0.1 to 1% (w / v).

[0075] In one embodiment of the invention, the stable liquid pharmaceutical formulation may be free of metal salts such as NaCl, KCl, NaF, KBr, NaBr, Na2SO4, NaSCN, and K2SO4. If the stable liquid pharmaceutical formulation contains these metal salts, precipitation may occur in the formulation, and the formulation may gel and may have poor stability.

[0076] In one embodiment of the invention, the stable liquid pharmaceutical formulation may be free of chelating agents (e.g., EDTA). If the pharmaceutical formulation contains chelating agents, its oxidation rate may be increased.

[0077] In one embodiment of the invention, the stable liquid pharmaceutical formulation may be preservative-free. Examples of preservatives include octadecyl dimethyl benzyl ammonium chloride, hexamethyl diammonium chloride, benzalkonium chloride, benzyl chloride, phenol, butanol, benzyl alcohol, alkyl p-hydroxybenzoate, catechol, resorcinol, cyclohexanol, 3-pentanol, m-cresol, etc. If the pharmaceutical formulation contains preservatives, the preservatives may not contribute to improving the stability of the pharmaceutical formulation.

[0078] In one embodiment of the invention, the stable liquid pharmaceutical formulation of the present invention may further contain additives known in the art that do not adversely affect the activity of the antibody and the stability and low viscosity of the formulation. For example, the pharmaceutical formulation may further contain an aqueous carrier, an antioxidant, or a mixture of two or more thereof. The aqueous carrier is a pharmaceutically acceptable carrier (safe and non-toxic when administered to humans) and can be used in the preparation of liquid pharmaceutical formulations. Examples of aqueous carriers include, but are not limited to, sterile water for injection (SWFI), bacteriostatic water for injection (BWFI), sterile saline solution, Ringer's solution, glucose, etc. Examples of antioxidants include, but are not limited to, ascorbic acid, etc.

[0079] (G) "Stable" liquid pharmaceutical preparations

[0080] The term "stable" in the context of "stable" liquid pharmaceutical formulations of this invention means that the antibody according to the invention substantially maintains its physical and / or chemical stability and / or biological activity during production and / or storage. Various analytical techniques for measuring protein stability are readily available in the art.

[0081] Physical stability can be assessed using methods known in the art, including measuring the apparent attenuation of light (absorbance or optical density) of the sample. This measurement of light attenuation is related to the turbidity of the formulation. Furthermore, for physical stability, the content of high molecular weight components, low molecular weight components, the amount of intact protein, the number of subvisible particles, etc., can be measured.

[0082] Chemical stability can be assessed, for example, by detecting and quantifying chemically altered forms of antibodies. Chemical stability includes charge changes (e.g., due to deamidation or oxidation), which can be evaluated, for example, by ion-exchange chromatography. For chemical stability, charge variations (acidic or basic peaks) can be measured.

[0083] Biological activity can be assessed using methods known in the art. For example, antigen-binding affinity can be measured using ELISA.

[0084] In one embodiment of the present invention, the liquid pharmaceutical preparation can be stable for a long time.

[0085] In one embodiment of the invention, the term "stable" liquid pharmaceutical preparation refers to a liquid pharmaceutical preparation that meets one or more of the following criteria.

[0086] Turbidity

[0087] - After storage at 40℃±2℃ for 4 weeks, it exhibits absorbance A as measured by a spectrophotometer. 600 Liquid pharmaceutical preparations with a concentration of 0 to 0.0300 or 0 to 0.0700;

[0088] - After storage under closed conditions at a temperature of 40℃±2℃ and a relative humidity of 75±5% for 4 weeks, it exhibits an absorbance A measured by a spectrophotometer. 600 Liquid pharmaceutical preparations with a concentration of 0 to 0.0300 or 0 to 0.0700;

[0089] Content of major components (main peak)

[0090] - A liquid pharmaceutical preparation whose main component content is 98% to 100% after storage at 40℃±2℃ for 4 weeks, as measured by SE-HPLC.

[0091] - A liquid pharmaceutical preparation whose main component content is 98% to 100% after storage for 4 weeks under closed conditions at a temperature of 40℃±2℃ and a relative humidity of 75±5%, as measured by SE-HPLC.

[0092] The content of high molecular weight components (the retention time of which is earlier than that of the main peak (intact IgG))

[0093] - A liquid pharmaceutical preparation with a high molecular weight component content of 0 to 1.00% after storage at 5℃±3℃ for 12 months, as measured by SE-HPLC.

[0094] - A liquid pharmaceutical preparation whose high molecular weight component content is 0 to 1.00% after storage at 5℃±3℃ for 12 months under closed conditions, as measured by SE-HPLC.

[0095] The content of low molecular weight components (whose retention time is later than that of the main peak (intact IgG))

[0096] - A liquid pharmaceutical preparation with a low molecular weight component content of 0 to 0.40% after storage at 5℃±3℃ for 12 months, as measured by SE-HPLC.

[0097] - A liquid pharmaceutical preparation with a low molecular weight component content of 0 to 0.40% after storage at 5℃±3℃ for 12 months under closed conditions, as measured by SE-HPLC.

[0098] Content of intact immunoglobulin G

[0099] - A liquid pharmaceutical preparation with an intact immunoglobulin G content (Intact IgG%) of 94.0% to 100% after storage at 5℃±3℃ for 12 months, as measured by non-reducing CE-SDS.

[0100] - A liquid pharmaceutical preparation with an intact immunoglobulin G content (Intact IgG%) of 94.0% to 100% after storage at 5℃±3℃ for 12 months under closed conditions, as measured by non-reducing CE-SDS.

[0101] - A liquid pharmaceutical preparation with an intact immunoglobulin G content (Intact IgG%) of 94.0% to 100% after storage at 40℃±2℃ for 4 weeks, as measured by non-reducing CE-SDS.

[0102] - A liquid pharmaceutical preparation with an intact immunoglobulin G content (Intact IgG%) of 94.0% to 100% after storage for 4 weeks under closed conditions at a temperature of 40℃±2℃ and a relative humidity of 75±5%, as measured by non-reducing CE-SDS.

[0103] Content of complete heavy and light chains

[0104] - The content of intact heavy and light chains (Intact HC+LC%) in liquid pharmaceutical preparations is 99.0% to 100% after storage at 5℃±3℃ for 12 months, as measured by reduced CE-SDS.

[0105] - A liquid pharmaceutical preparation with an intact heavy chain and light chain content (Intact HC+LC%) of 99.0% to 100% after storage for 12 months under closed conditions at a temperature of 5℃±3℃, as measured by reduced CE-SDS.

[0106] - A liquid pharmaceutical preparation with an intact heavy chain and light chain content (Intact HC+LC%) of 98.0% to 100% after storage at 40℃±2℃ for 4 weeks, as measured by reduced CE-SDS.

[0107] -Measured by reduced CE-SDS, after 4 weeks of storage under closed conditions at a temperature of 40℃±2℃ and a relative humidity of 75±5%, the content of intact heavy and light chains (Intact HC+LC%) is 98.0% to 100% in liquid pharmaceutical preparations.

[0108] Number of subvisible particles

[0109] - Liquid pharmaceutical preparations with a subvisible particle count (≥10.00μm, <400.00μm) of 0 to 1,000 after storage at 5℃±3℃ for 12 months, as measured by HIAC.

[0110] - Liquid pharmaceutical preparations with a number of subvisible particles (≥10.00μm, <400.00μm) of 0 to 1,000 after storage at 5℃±3℃ for 12 months under closed conditions, as measured by HIAC.

[0111] - Liquid pharmaceutical preparations with a subvisible particle count (≥1.00μm, <100.00μm) of 0 to 30,000 after storage at 40℃±2℃ for 4 weeks, as measured by MFI;

[0112] - Liquid pharmaceutical formulations with a subvisible particle count (≥1.00μm, <100.00μm) of 0 to 30,000 after storage for 4 weeks under closed conditions at a temperature of 40℃±2℃ and a relative humidity of 75±5%; as measured by MFI.

[0113] - Liquid pharmaceutical preparations with a subvisible particle count (≥10.00μm, <100.00μm) of 0 to 200 after storage at 40℃±2℃ for 4 weeks, as measured by MFI;

[0114] - Liquid pharmaceutical formulations with a subvisible particle count (≥10.00μm, <100.00μm) of 0 to 200 after storage for 4 weeks under closed conditions at a temperature of 40℃±2℃ and a relative humidity of 75±5%; as measured by MFI.

[0115] - Liquid pharmaceutical preparations with a subvisible particle count (≥10.00μm, <100.00μm) of 0 to 500 after storage at 40℃±2℃ for 6 weeks, as measured by MFI;

[0116] - Liquid pharmaceutical formulations with a subvisible particle count (≥10.00μm, <100.00μm) of 0 to 500 after storage for 6 weeks under closed conditions at a temperature of 40℃±2℃ and a relative humidity of 75±5%; as measured by MFI.

[0117] Oxidation rate

[0118] - Liquid pharmaceutical formulations with an oxidation rate of heavy chain Met 255 of 0% to 2.5% after storage at 40℃±2℃ for 4 weeks, as measured by LC-MS;

[0119] - Liquid pharmaceutical formulations with an oxidation rate of 0% to 2.5% of heavy chain Met 255 after storage for 4 weeks under closed conditions at a temperature of 40℃±2℃ and a relative humidity of 75±5%, as measured by LC-MS.

[0120] Charge variation

[0121] - Liquid pharmaceutical preparations that show 20% to 35% acidic peaks after being stored at 40℃±2℃ for 4 weeks, as measured by IEC-HPLC;

[0122] - A liquid pharmaceutical preparation that, after being stored under closed conditions at a temperature of 40℃±2℃ and a relative humidity of 75±5% for 4 weeks, shows an acidic peak of 20% to 35% as measured by IEC-HPLC.

[0123] - A liquid pharmaceutical preparation that, after being stored at 40℃±2℃ for 4 weeks, shows an alkaline peak of 33% to 40% as measured by IEC-HPLC;

[0124] - A liquid pharmaceutical preparation that, after being stored under closed conditions at a temperature of 40℃±2℃ and a relative humidity of 75±5% for 4 weeks, shows an alkaline peak of 33% to 40% as measured by IEC-HPLC.

[0125] TNF-α binding affinity

[0126] - Liquid pharmaceutical formulations exhibiting 80% to 120% TNF-α binding affinity as measured by ELISA after storage at 5℃±3℃ for 12 months; and

[0127] - A liquid pharmaceutical preparation with 80% to 120% TNF-α binding affinity as measured by ELISA after being stored for 12 months under closed conditions at a temperature of 5℃±3℃.

[0128] In one embodiment of the invention, the viscosity of the pharmaceutical preparation after storage at 40°C ± 2°C for one month can be between 0.5 cp and 10.0 cp. In another embodiment of the invention, the viscosity of the pharmaceutical preparation after storage at 5°C ± 3°C for six months can be between 0.5 cp and 5.0 cp.

[0129] [Methods for preparing stable liquid pharmaceutical formulations]

[0130] The stable liquid pharmaceutical formulation of the present invention can be prepared using any known method, and the method is not limited to a specific method. For example, a stable liquid pharmaceutical formulation can be prepared by adding a buffer to a solution containing a surfactant and a sugar or a derivative thereof, while adjusting the pH of the solution, and then adding an antibody to the mixed solution. Alternatively, a liquid pharmaceutical formulation can be prepared by preparing a solution containing some excipients in the final step of the purification process, and then adding the remaining components to the solution. For example, a liquid pharmaceutical formulation can be prepared by preparing a solution containing an antibody, a buffer, and a sugar or a derivative thereof, and then adding a surfactant to the solution.

[0131] In addition, the preparation method of the formulation may or may not include a freeze-drying step.

[0132] When the preparation method does not include a freeze-drying step, for example, the liquid pharmaceutical preparation prepared according to the present invention can be sterilized and then immediately placed in a closed container.

[0133] When the preparation method includes a freeze-drying step, for example, the liquid pharmaceutical formulation prepared according to the present invention can be freeze-dried or freeze-dried and stored, and then the components that have been removed or modified can be replenished or replaced by freeze-drying and / or storage, thereby preparing the liquid pharmaceutical formulation according to the present invention. Alternatively, except for the components that have been removed or modified by freeze-drying and / or storage, only the components of the liquid pharmaceutical formulation of the present invention can be freeze-dried or freeze-dried and stored, and then the excluded components can be added, thereby preparing the liquid pharmaceutical formulation according to the present invention.

[0134] [Methods for using stable liquid pharmaceutical formulations]

[0135] The stable liquid pharmaceutical formulation according to the present invention can be used to treat diseases in which TNF-α activity is a harmful factor. Examples of diseases in which TNF-α activity is a harmful factor include, but are not limited to, sepsis, autoimmune diseases, infectious diseases, transplant rejection, malignant cancers, lung diseases, enteropathic diseases, and heart diseases.

[0136] In one embodiment of the present invention, diseases in which TNF-α activity is a harmful factor can be selected from rheumatoid arthritis, ankylosing spondylitis, ulcerative colitis, adult Crohn's disease, pediatric Crohn's disease, psoriasis, and psoriatic arthritis.

[0137] The stable liquid pharmaceutical formulation according to the invention can be provided as a single-dose form, a multi-dose form, or a form for subcutaneous autoinjection.

[0138] The concentrations of other components (including antibodies) in the liquid drug formulation are as described above, and the total volume of the liquid drug formulation may be from 0.2 to 2.0 mL.

[0139] The dosage and timing of liquid drug formulations can vary depending on the type, severity, and course of the disease, the patient's health and response to treatment, and the judgment of the treating physician, and are not limited to specific dosages and times. For example, based on antibody concentration, one or more products containing liquid drug formulations can be administered at a dose of 1 to 10 mg / kg, followed by the same or different doses at intervals of one week, two weeks, three weeks, one month, two months, or three months.

[0140] In one embodiment of the invention, a stable liquid pharmaceutical formulation may not undergo a reconstitution step, a dilution step, or both before use.

[0141] [Treatment methods and stabilization techniques]

[0142] The present invention also provides a method for treating a patient with a disease in which TNF-α activity is a harmful factor, the method comprising administering to the patient a stable liquid pharmaceutical preparation containing: (A) an antibody or an antigen-binding fragment thereof; (B) a surfactant; (C) a sugar or a derivative thereof; and (D) a buffer region.

[0143] The present invention also provides a method for stabilizing antibodies in a liquid pharmaceutical preparation, the method comprising preparing a stable liquid pharmaceutical preparation containing: (A) an antibody or an antigen-binding fragment thereof; (B) a surfactant; (C) a sugar or a derivative thereof; and (D) a buffer.

[0144] In one implementation of the treatment or stabilization method, antibody (A) may comprise an antibody that binds to TNF-α.

[0145] In one embodiment of the treatment or stabilization method, antibody (A) may comprise infliximab, adalimumab, PEGylated cetrus Fab fragment, golimumab, or a mixture thereof.

[0146] In one implementation of a treatment or stabilization method, antibody (A) may comprise a chimeric human-mouse IgG monoclonal antibody.

[0147] In one embodiment of the treatment or stabilization method, the antibody (A) or its antigen-binding fragment may comprise: a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises a CDR1 domain containing the amino acid sequence shown in SEQ ID NO: 1, a CDR2 domain containing the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 domain containing the amino acid sequence shown in SEQ ID NO: 3; and the heavy chain variable region comprises a CDR1 domain containing the amino acid sequence shown in SEQ ID NO: 4, a CDR2 domain containing the amino acid sequence shown in SEQ ID NO: 5, and a CDR3 domain containing the amino acid sequence shown in SEQ ID NO: 6.

[0148] In one embodiment of the treatment or stabilization method, the antibody or its antigen-binding fragment (A) may comprise: a light chain variable region having the amino acid sequence shown in SEQ ID NO: 7; and a heavy chain variable region having the amino acid sequence shown in SEQ ID NO: 8.

[0149] In one embodiment of the treatment or stabilization method, the antibody (A) may comprise: a light chain having the amino acid sequence shown in SEQ ID NO: 9; and a heavy chain having the amino acid sequence shown in SEQ ID NO: 10.

[0150] In one embodiment of the treatment or stabilization method, the concentration of the antibody or its antigen-binding fragment (A) may be 10 to 200 mg / ml.

[0151] In one embodiment of the treatment or stabilization method, surfactant (B) may comprise polysorbate, poloxamer, or a mixture thereof.

[0152] In one embodiment of the treatment or stabilization method, the surfactant (B) may comprise polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, or a mixture of two or more thereof.

[0153] In one embodiment of the treatment or stabilization method, surfactant (B) may comprise polysorbate 80.

[0154] In one embodiment of the treatment or stabilization method, the concentration of surfactant (B) may be 0.02 to 0.1% (w / v).

[0155] In one embodiment of the treatment or stabilization method, the sugar (C) may comprise monosaccharides, disaccharides, oligosaccharides, polysaccharides, or mixtures of two or more thereof, and the sugar derivative (C) may comprise sugar alcohols, sugar acids, or mixtures thereof.

[0156] In one embodiment of the treatment or stabilization method, the sugar or its derivative (C) may comprise sorbitol, mannitol, trehalose, sucrose, or a mixture of two or more thereof.

[0157] In one embodiment of the treatment or stabilization method, the concentration of the sugar or its derivative (C) may be 1 to 10% (w / v).

[0158] In one implementation of the treatment or stabilization method, the buffer (D) may contain acetate or histidine.

[0159] In one embodiment of the treatment or stabilization method, the buffer (D) may have a concentration of 1 to 50 mM.

[0160] In one embodiment of the treatment or stabilization method, the stable liquid pharmaceutical formulation may have a pH of 4.0 to 5.5.

[0161] In one embodiment of the treatment or stabilization method, the stabilized liquid pharmaceutical formulation may be free of aspartic acid, lysine, arginine, or mixtures thereof.

[0162] In one embodiment of the treatment or stabilization method, the stabilized liquid pharmaceutical preparation may be free of NaCl, KCl, NaF, KBr, NaBr, Na2SO4, NaSCN, K2SO4, or mixtures thereof.

[0163] In one implementation of a treatment or stabilization method, the stabilized liquid pharmaceutical formulation may be free of chelating agents.

[0164] In one implementation of a treatment or stabilization method, the stabilized liquid pharmaceutical formulation may be preservative-free.

[0165] In one embodiment of the treatment or stabilization method, the stabilized liquid pharmaceutical formulation may further contain an aqueous carrier, an antioxidant, or a mixture of two or more thereof.

[0166] In one embodiment of the treatment or stabilization method, the viscosity of the stabilized liquid pharmaceutical preparation measured after storage at 40°C ± 2°C for 1 month may be 0.5 cp to 10 cp, or the viscosity measured after storage at 5°C ± 3°C for 6 months may be 0.5 cp to 5 cp.

[0167] In one embodiment of the treatment or stabilization method, the stabilized liquid pharmaceutical formulation may contain: (A) an antibody or an antigen-binding fragment thereof comprising a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises a CDR1 domain containing the amino acid sequence shown in SEQ ID NO: 1, a CDR2 domain containing the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 domain containing the amino acid sequence shown in SEQ ID NO: 3; the heavy chain variable region comprises a CDR1 domain containing the amino acid sequence shown in SEQ ID NO: 4, a CDR2 domain containing the amino acid sequence shown in SEQ ID NO: 5, and a CDR3 domain containing the amino acid sequence shown in SEQ ID NO: 6; (B) a surfactant; (C) a sugar or a derivative thereof; and (D) a buffer containing acetate or histidine.

[0168] In one embodiment of the treatment or stabilization method, the stabilized liquid pharmaceutical formulation may contain: (A) 90 to 145 mg / ml of an antibody or an antigen-binding fragment thereof comprising a light chain variable region and a heavy chain variable region, wherein the light chain variable region comprises a CDR1 domain containing the amino acid sequence shown in SEQ ID NO: 1, a CDR2 domain containing the amino acid sequence shown in SEQ ID NO: 2, and a CDR3 domain containing the amino acid sequence shown in SEQ ID NO: 3; the heavy chain variable region comprises a CDR1 domain containing the amino acid sequence shown in SEQ ID NO: 4, a CDR2 domain containing the amino acid sequence shown in SEQ ID NO: 5, and a CDR3 domain containing the amino acid sequence shown in SEQ ID NO: 6; (B) 0.02 to 0.1% (w / v) of a surfactant; (C) 1 to 10% (w / v) of a sugar or a derivative thereof; and (D) 1 to 50 mM of a buffer containing acetate or histidine.

[0169] In one implementation of the treatment method, a stable liquid pharmaceutical preparation may be administered subcutaneously.

[0170] In one implementation of a treatment or stabilization method, a stable liquid pharmaceutical preparation may not undergo a reconstitution step, a dilution step, or both before use.

[0171] In one implementation of a treatment or stabilization method, the stabilized liquid pharmaceutical preparation may be filled into a pre-filled syringe before use.

[0172] In one implementation of a treatment or stabilization method, a pre-filled syringe may be included in an auto-injector.

[0173] [product]

[0174] The present invention also provides a product comprising: a stable liquid pharmaceutical preparation; and a container that receives the stable liquid pharmaceutical preparation in a closed state.

[0175] Stable liquid pharmaceutical formulations are as described above.

[0176] In one embodiment of the invention, the container may be formed of materials such as glass, polymer (plastic), metal, etc., but is not limited thereto. In one embodiment of the invention, the container is a bottle, vial, injection cartridge, syringe (pre-filled syringe, auto-injector), or tube, but is not limited thereto. In one embodiment of the invention, the container may be a glass or polymer vial, or a glass or polymer pre-filled syringe.

[0177] The specific product forms of the vials, injection cartridges, pre-filled syringes, or autoinjectors described above, as well as the methods for filling stable liquid pharmaceutical formulations into the vials, injection cartridges, pre-filled syringes, or autoinjectors, are readily available or implementable by any person skilled in the art to which this invention pertains. For example, U.S. Patent Nos. 4,861,335 and 6,331,174 disclose specific product forms and filling methods for pre-filled syringes. For example, U.S. Patent Nos. 5,085,642 and 5,681,291 disclose specific product forms and assembly methods for autoinjectors. The vials, injection cartridges, pre-filled syringes, or autoinjectors used in this invention can be commercially available products or products manufactured separately taking into account the physical properties of stable liquid pharmaceutical formulations, the area of ​​application of the formulation, the dosage of the formulation, etc.

[0178] In one embodiment of the invention, the interior of the container may not be coated with silicone oil. Coating with silicone oil reduces the stability of the formulation. The container may be a single-dose or multi-dose container.

[0179] In one embodiment of the invention, the product may further include instructions providing a method of using a stable liquid pharmaceutical formulation, a method of storing the formulation, or both. Methods of using the formulation include methods of treating diseases in which TNF-α activity is a harmful factor, and may include the route of administration, dosage, and timing of administration.

[0180] In one embodiment of the invention, the product may include other necessary instruments (e.g., needles, syringes, etc.) from both a business and user perspective.

[0181] The invention will be described below with reference to embodiments. However, it should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention.

[0182] Example

[0183] The antibody used in the following experimental examples is infliximab, which was purified from commercially available Remsima (manufactured by Celltrion).

[0184] The physical stability, chemical stability, and biological activity of the liquid pharmaceutical preparations used in the following experimental examples were measured using the following methods.

[0185] - Turbidity

[0186] The absorbance at 600 nm was measured using a UV-Vis spectrophotometer.

[0187] - Content of main components

[0188] The content of the main component (main peak; %) was determined using size exclusion HPLC.

[0189] - Content of high molecular weight components

[0190] The content (pre-peak; %) of high molecular weight components was determined using size exclusion HPLC.

[0191] - Content of low molecular weight components

[0192] The content (post-peak; %) of low molecular weight components was determined using size exclusion HPLC.

[0193] - Content of intact immunoglobulin G (Intact IgG%)

[0194] The percentage of intact immunoglobulin G was measured using non-reduced capillary electrophoresis-sodium dodecyl sulfate (NRCE-SDS).

[0195] - Content of intact heavy and light chains (Intact HC+LC%)

[0196] The percentage of intact heavy and light chains was determined using reduced capillary electrophoresis-sodium dodecyl sulfate (RCE-SDS).

[0197] - The number of sub-visible particles

[0198] Experiments 1 to 4: Using microflow imaging (MFI) to measure the number of subvisible particles.

[0199] Experimental Example 5: The number of subvisible particles was measured using a light-shielding liquid particle counter (model: HIAC 9703).

[0200] -Oxidation

[0201] The oxidation (%) of heavy chain Met 255 was measured by peptide mapping using liquid chromatography-mass spectrometry (LC-MS).

[0202] - Charge variation

[0203] Acid and basic peaks (%) were measured by ion exchange chromatography-high performance liquid chromatography (IEC-HPLC).

[0204] -TNF-α binding affinity

[0205] TNF-α binding affinity (%) was measured by enzyme-linked immunosorbent assay (ELISA).

[0206] - viscosity

[0207] A microcapillary flow system equipped with a flow cell (B05 sensor type; 50 μm cell depth) was used (apparent shear rate: 10). 3 Up to 10 5 s -1 Viscosity was measured in a 500 μL syringe at 25 °C ± 0.1 °C.

[0208] Experimental Example 1: Comparison of sugar alcohols and NaCl; Comparison of acetate / histidine buffers and citrate / phosphate buffers; Comparison of pH 4-5.5 and pH 6-7.

[0209] To prepare the liquid pharmaceutical formulation for Experimental Example 1, each buffer was prepared to have the desired pH, and sorbitol or NaCl was added to it. Then, antibodies and surfactants were added to prepare the samples shown in Table 1 below. The specific amounts of each component are shown in Table 1 below. The concentration of the buffer refers to the molecular / anion concentration of the corresponding compound. The total volume is 1 ml.

[0210] [Table 1]

[0211]

[0212]

[0213] Liquid pharmaceutical formulations prepared according to Experimental Examples 1 to 3 and Comparative Examples 1 to 8 were stored for 2 weeks at a temperature of 40 ± 2 °C and a relative humidity of 75 ± 5%. The results showed that the formulations containing NaCl (Comparative Examples 1, 2, 4, 5, and 7) all exhibited a precipitate and a gel-like form. Furthermore, Comparative Example 3, containing sorbitol but also sodium citrate, and Comparative Example 8, containing sorbitol but also sodium phosphate, also exhibited a gel-like form.

[0214] Among the formulations containing sorbitol, only the formulations of Experimental Examples 1, 2, and 3, and Comparative Example 6, did not exhibit a gel form. The stability of the formulations was measured after storage at 5 ± 3 °C for 0, 2, and 4 weeks, and after storage at 40 ± 2 °C and 75 ± 5% relative humidity for 2 and 4 weeks. The results are shown in Tables 2 to 9 below.

[0215] Turbidity

[0216] [Table 2]

[0217]

[0218] As can be seen from Table 2 above, the formulation of Experimental Example 1, with a pH of 4 and containing acetate as a buffer, exhibits the best turbidity, especially after storage at 40°C for 4 weeks, showing an absorbance of 0.0300 or lower. Furthermore, it can be seen that the formulations of Experimental Examples 2 and 3, with a pH of 5.5 and containing histidine as a buffer, also show absorbance of 0.0700 or lower after storage at 40°C for 4 weeks.

[0219] However, it can be seen that the formulation of Comparative Example 6, which has a pH of 6 and contains phosphate as a buffer, showed a significant increase in turbidity after being stored at 40°C for 2 and 4 weeks.

[0220] Content of high molecular weight components

[0221] [Table 3]

[0222]

[0223] As can be seen from Table 3 above, the formulation of Experimental Example 1 showed the lowest content of high molecular weight components under all conditions. In particular, the formulation of Experimental Example 1 showed a high molecular weight component content of 1.0% or less after being stored at 40°C for 4 weeks. Furthermore, it can be seen that the formulations of Experimental Examples 2 and 3 showed a high molecular weight component content of 1.5% or less after being stored at 40°C for 4 weeks.

[0224] Content of intact immunoglobulin G (Intact IgG%)

[0225] [Table 4]

[0226]

[0227] As can be seen from Table 4 above, after storage at 40°C for 4 weeks, the content of intact immunoglobulins in the preparations of Experimental Examples 1 to 3 was 94.0% or higher, which was higher than that in Comparative Example 6.

[0228] Content of intact heavy and light chains (Intact HC+LC%)

[0229] [Table 5]

[0230]

[0231] As can be seen from Table 5 above, after being stored at 40°C for 4 weeks, the content of intact heavy and light chains in the formulations of Experimental Examples 1 to 3 was 98.0% or higher, which was higher than that in Comparative Example 6.

[0232] Oxidation rate (heavy chain Met 255)

[0233] [Table 6]

[0234] After 0 weeks at 40±2℃ After 4 weeks at 40±2℃ Experimental Example 1 2.2 2.4 Experimental Example 2 2.0 2.5 Experimental Example 3 2.1 2.5 Comparative Example 6 2.2 4.1

[0235] As can be seen from Table 6 above, after storage at 40°C for 4 weeks, the oxidation rate of heavy chain Met255 in the formulations of Experimental Examples 1 to 3 was 2.5% or lower, which was lower than the oxidation rate of Comparative Example 6.

[0236] Charge variation (acid peak)

[0237] [Table 7]

[0238]

[0239] As can be seen from Table 7, after storage at 40°C for 4 weeks, the acid peaks of the formulations in Experimental Examples 1 to 3 were 35% or lower, which was lower than that of Comparative Example 6. This indicates that the formulations in Experimental Examples 1 to 3 are stable formulations, with less deamidation, which is the main cause of the increase in acid peaks.

[0240] Charge variation (basic peak)

[0241] [Table 8]

[0242]

[0243] As can be seen from Table 8 above, after being stored at 40°C for 4 weeks, the alkaline peak of the preparations in Experimental Examples 1 to 3 was 33% or higher, which was higher than that of Comparative Example 6.

[0244] Number of subvisible particles (≥1.00 μm, <100.00 μm)

[0245] [Table 9]

[0246]

[0247] As can be seen from Table 9, after storage at 40°C for 4 weeks, the number of subvisible particles (≥1.00 μm, <100.00 μm) in the formulations of Experimental Examples 1 to 3 was 30,000 or less, which was lower than that in Comparative Example 6.

[0248] Experimental Example 2: The Role of Amino Acids

[0249] To prepare the liquid pharmaceutical formulation for Experimental Example 2, a buffer containing sodium acetate was prepared to have the desired pH, and sorbitol was added thereto. Then, an antibody, along with a surfactant and amino acid / taurine, were added to prepare the sample shown in Table 10 below. The concentrations of each component are shown in Table 10 below. The concentration of the buffer refers to the concentration of acetate anions. The total volume is 1 ml.

[0250] [Table 10]

[0251]

[0252]

[0253] 1) Add amino acids or taurine at 5% (w / v) or less.

[0254] The formulations containing aspartic acid, histidine, lysine and arginine, respectively, in comparative ratios 9, 10, 11 and 12, became solid after being stored at 50±2℃ for 24 hours.

[0255] For formulations containing other amino acids or taurine, stability was measured after storage at 5±3°C and 50±2°C for 24 hours, but there were no significant differences between these formulations or between these formulations and the formulation of Experimental Example 1.

[0256] Experimental Example 3: Protein concentration; surfactant concentration; and type of sugar.

[0257] To prepare the liquid pharmaceutical formulation for Experimental Example 3, a buffer containing sodium acetate was prepared to have the desired pH, and sorbitol, mannitol, trehalose, or sucrose was added thereto. Then, an antibody and a surfactant were added to prepare the sample shown in Table 11 below. The concentrations of each component are shown in Table 11 below. The concentration of the buffer refers to the concentration of acetate anions. The total volume is 1 ml.

[0258] [Table 11]

[0259]

[0260] The stability of the formulation was measured after storage at 5±3℃ for 0, 2, and 4 weeks, and after storage at 40±2℃ and 75±5% relative humidity for 2 and 4 weeks. The results are shown in Tables 12 to 17 below.

[0261] protein concentration

[0262] Content of high molecular weight components

[0263] [Table 12]

[0264]

[0265] As can be seen from Table 12 above, the content of high molecular weight components increases with increasing antibody concentration. However, when the antibody concentration ranges from 90 to 145 mg / ml, the content of high molecular weight components is generally low after storage at 5°C and 40°C for 4 weeks.

[0266] surfactant concentration

[0267] Number of subvisible particles (≥1.00 μm, <100.00 μm)

[0268] [Table 13]

[0269]

[0270] As can be seen from Table 13 above, at surfactant concentrations ranging from 0.02% to 0.1% (w / v), after storage at 40°C for 4 weeks, the number of subvisible particles (≥1.00 μm, <100.00 μm) is 10,000 or less.

[0271] Types of sugar

[0272] Content of main peak

[0273] [Table 14]

[0274]

[0275] As can be seen from Table 14 above, preparations containing sorbitol, mannitol, trehalose or sucrose as sugars showed a content of 98% or higher of the main components after being stored at 40°C for 4 weeks.

[0276] Charge variation (acid peak)

[0277] [Table 15]

[0278]

[0279] As can be seen from Table 15 above, preparations containing sorbitol, mannitol, trehalose or sucrose as sugars showed an acid peak of 35% or lower after being stored at 40°C for 4 weeks.

[0280] Number of subvisible particles (≥1.00 μm, <100.00 μm)

[0281] [Table 16]

[0282]

[0283] Number of subvisible particles (≥10.00 μm, <100.00 μm)

[0284] [Table 17]

[0285]

[0286] As can be seen in Tables 16 and 17 above, in formulations containing sorbitol, mannitol, trehalose, or sucrose as sugars, the number of subvisible particles (≥1.00 μm, <100.00 μm) after 4 weeks of storage at 40°C was 15,000 or less, and the number of subvisible particles (≥10.00 μm, <100.00 μm) after 4 weeks of storage at 40°C was 200 or less.

[0287] Experiment Example 4: Types of Surfactants and the Role of Chelating Agents

[0288] To prepare the liquid pharmaceutical formulation for Experimental Example 4, a buffer containing sodium acetate was prepared to have the desired pH, and sorbitol was added thereto. Then, an antibody was added, along with a surfactant or a mixture of surfactant and chelating agent, to prepare the sample shown in Table 18 below. The content of each component is shown in Table 18 below. The concentration of the buffer refers to the concentration of acetate anions. The total volume is 1 ml.

[0289] [Table 18]

[0290]

[0291] The stability of the formulations shown in Table 18 above was measured after 0, 3, and 6 weeks of storage at 5±3℃, 25±2℃ and 60±5% relative humidity, and under closed conditions at 40±2℃ and 75±5% relative humidity. The measurement results are shown in Tables 19 and 20 below.

[0292] Types of surfactants

[0293] Number of subvisible particles (≥10.00 μm, <100.00 μm)

[0294] [Table 19]

[0295]

[0296] As can be seen from Table 19 above, in the formulation of Experimental Example 13 containing polysorbate 80 as a surfactant, after storage at 40°C for 6 weeks, the number of subvisible particles (≥10.00 μm, <100.00 μm) was 100 or less (the lowest), and in the formulation of Experimental Example 15 containing poloxamer 188 as a surfactant, after storage at 40°C for 6 weeks, the number of subvisible particles (≥10.00 μm, <100.00 μm) was 2000 or more (the highest).

[0297] The role of chelating agents (EDTA)

[0298] Oxidation rate (heavy chain Met 255)

[0299] [Table 20]

[0300]

[0301] As can be seen from Table 20 above, compared with the formulations of Experimental Examples 13 to 15 which do not contain chelating agent (EDTA), the oxidation rate of heavy chain Met 255 increased after storage at 40°C for 6 weeks.

[0302] Experimental Example 5: Long-term stability

[0303] To prepare the liquid pharmaceutical formulation for Experimental Example 5, a buffer containing sodium acetate was prepared to a pH of 5.0, and sorbitol was added to it. Then, an antibody and a surfactant were added to prepare the sample shown in Table 21 below. The concentrations of each component are shown in Table 21 below. The concentration of the buffer refers to the concentration of acetate anions. The total volume is 1 ml.

[0304] [Table 21]

[0305]

[0306] The stability of the formulations shown in Table 21 was measured after storage at 5 ± 3 °C for 0, 3, and 6 months under closed conditions. The measurement results are shown in Tables 22 to 27 below.

[0307] Number of subvisible particles (≥10.00μm, <400.00μm)

[0308] [Table 22]

[0309]

[0310] As can be seen from Table 22 above, after 12 months of storage at 5°C, the number of subvisible particles (≥10.00 μm, <400.00 μm) in the formulation of Experimental Example 16 was as low as 100 or less.

[0311] Content of intact immunoglobulins (Intact IgG%)

[0312] [Table 23]

[0313]

[0314] As can be seen from Table 23 above, after being stored at 5°C for 12 months, the content of intact immunoglobulin G in the preparation of Experimental Example 16 was as high as 94% or higher.

[0315] Content of intact heavy and light chains (Intact HC+LC%)

[0316] [Table 24]

[0317]

[0318] As can be seen from Table 24 above, after being stored at 5°C for 12 months, the content of intact heavy and light chains in the formulation of Experimental Example 16 was as high as 99% or higher.

[0319] Content of high molecular weight components

[0320] [Table 25]

[0321]

[0322] As can be seen from Table 25 above, after being stored at 5°C for 12 months, the content of high molecular weight components in the formulation of Experimental Example 16 was as low as 1.0% or less.

[0323] Content of low molecular weight components

[0324] [Table 26]

[0325]

[0326] As can be seen from Table 26 above, after being stored at 5°C for 12 months, the content of low molecular weight components in the formulation of Experimental Example 16 was as low as 0.4% or less.

[0327] TNF-α binding affinity

[0328] [Table 27]

[0329]

[0330] As can be seen from Table 27 above, after being stored at 5°C for 12 months, the TNF-α binding affinity of the formulation in Experimental Example 16 was as high as 95% or higher.

[0331] The viscosity of the formulation of Experimental Example 16 was measured after storage at 40±2℃ for 0, 0.5, 1, 2, and 3 months under sealed conditions, and after storage at 5±3℃ for 6 months under sealed conditions. The measurement results are shown in Table 28 below.

[0332] Viscosity (cP)

[0333] [Table 28]

[0334]

[0335] As can be seen from Table 28 above, after being stored at 40℃±2℃ for 1 month, the viscosity of the formulation in Experimental Example 16 remained at a low level (8.0 cp), and after being stored at 5℃±3℃ for 6 months, it remained at a low level (4.0 cp). <110> Certified Co., Ltd. <120> Stable liquid pharmaceutical preparations <130> CPP2017022PCT <150> KR 10-2016-0083039 <151> 2016-06-30 <160> 10 <170> KoPatentIn 3.0 <210> 1 <211> 6 <212> PRT <213> Artificial sequence <220> <223> Antibody <400> 1 Gln Phe Val Gly Ser Ser 1 5 <210> 2 <211> 3 <212> PRT <213> Artificial sequence <220> <223> Antibody <400> 2 Tyr Ala Ser 1 <210> 3 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Antibody <400> 3 Gln Gln Ser His Ser Trp Pro Phe Thr 1 5 <210> 4 <211> 8 <212> PRT <213> Artificial sequence <220> <223> Antibody <400> 4 Gly Phe Ile Phe Ser Asn His Trp 1 5 <210> 5 <211> 10 <212> PRT <213> Artificial sequence <220> <223> Antibody <400> 5 Ile Arg Ser Lys Ser Ile Asn Ser Ala Thr 1 5 10 <210> 6 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Antibody <400> 6 Ser Arg Asn Tyr Tyr Gly Ser Thr Tyr Asp Tyr 1 5 10 <210> 7 <211> 107 <212> PRT <213> artificial sequence <220> <223> antibody <400> 7 Asp Ile Leu Leu Thr Gln Ser Pro Ala Ile Leu Ser Val Ser Pro Gly 1 5 10 15 Glu Arg Val Ser Phe Ser Cys Arg Ala Ser Gln Phe Val Gly Ser Ser 20 25 30 Ile His Trp Tyr Gln Gln Arg Thr Asn Gly Ser Pro Arg Leu Leu Ile 35 40 45 Lys Tyr Ala Ser Glu Ser Met Ser Gly Ile Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Ser Ile Asn Thr Val Glu Ser 65 70 75 80 Glu Asp Ile Ala Asp Tyr Tyr Cys Gln Gln Ser His Ser Trp Pro Phe 85 90 95 Thr Phe Gly Ser Gly Thr Asn Leu Glu Val Lys 100 105 <210> 8 <211> 119 <212> PRT <213> artificial sequence <220> <223> antibody <400> 8 Glu Val Lys Leu Glu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Met Lys Leu Ser Cys Val Ala Ser Gly Phe Ile Phe Ser Asn His 20 25 30 Trp Met Asn Trp Val Arg Gln Ser Pro Glu Lys Gly Leu Glu Trp Val 35 40 45 Ala Glu Ile Arg Ser Lys Ser Ile Asn Ser Ala Thr His Tyr Ala Glu 50 55 60 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Ala 65 70 75 80 Val Tyr Leu Gln Met Thr Asp Leu Arg Thr Glu Asp Thr Gly Val Tyr 85 90 95 Tyr Cys Ser Arg Asn Tyr Tyr Gly Ser Thr Tyr Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Leu Thr Val Ser 115 <210> 9 <211> 236 <212> PRT <213> Artificial Sequence <220> <223> Antibody <400> 9 Met Asp Phe Gln Val Gln Ile Ile Ser Phe Leu Leu Ile Ser Ala Ser 1 5 10 15 Val Ile Met Ser Arg Gly Asp Ile Leu Leu Thr Gln Ser Pro Ala Ile 20 25 30 Leu Ser Val Ser Pro Gly Glu Arg Val Ser Phe Ser Cys Arg Ala Ser 35 40 45 Gln Phe Val Gly Ser Ser Ile His Trp Tyr Gln Gln Arg Thr Asn Gly 50 55 60 Ser Pro Arg Leu Leu Ile Lys Tyr Ala Ser Glu Ser Met Ser Gly Ile 65 70 75 80 Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Ser 85 90 95 Ile Asn Thr Val Glu Ser Glu Asp Ile Ala Asp Tyr Tyr Cys Gln Gln 100 105 110 Ser His Ser Trp Pro Phe Thr Phe Gly Ser Gly Thr Asn Leu Glu Val 115 120 125 Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp 130 135 140 Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn 145 150 155 160 Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu 165 170 175 Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp 180 185 190 Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr 195 200 205 Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser 210 215 220 Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 235 <210> 10 <211> 469 <212> PRT <213> Artificial Sequence <220> <223> Antibody <400> 10 Met Gly Trp Ser Leu Ile Leu Leu Phe Leu Val Ala Val Ala Thr Arg 1 5 10 1​​​​​​​​​​​​​​​65 70 75 80 Tyr Ala Glu Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser 85 90 95 Lys Ser Ala Val Tyr Leu Gln Met Thr Asp Leu Arg Thr Glu Asp Thr 100 105 110 Gly Val Tyr Tyr Cys Ser Arg Asn Tyr Tyr Gly Ser Thr Tyr Asp Tyr 115 120 125 Trp Gly Gln Gly Thr Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly 130 135 140 Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly 145 150 155 160 Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val 165 170 175 Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe 180 185 190 Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val 195 200 205 Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val 210 215 220 Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys 225 230 235 240 Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu 245 250 255 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 260 265 270 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 275 280 285 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 290 295 300 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 305 310 315 320 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 325 330 335 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 340 345 350 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 355 360 365 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln 370 375 380 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 385 390 395 400 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 405 410 415 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 420 425 430 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 435 440 445 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 450 455 460 Leu Ser Pro Gly Lys 465

Claims

1. A stable liquid pharmaceutical preparation comprising: 90 to 145 mg / ml infliximab; 0.02% to 0.1% w / v surfactant; 1% to 10% w / v sorbitol, mannitol, trehalose, sucrose or one or more of these; and 1 mM to 50 mM buffers containing acetate; in, The pH of the formulation is 4.0 to 5.5; The formulation does not contain: i) Aspartic acid, lysine, arginine, histidine, or a mixture thereof; ii) NaCl, KCl, NaF, KBr, NaBr, Na2SO4, NaSCN, K2SO4, or mixtures thereof; and iii) Chelating agents; The surfactant mentioned therein is polysorbate, poloxamer, or a mixture thereof.

2. The stable liquid pharmaceutical formulation according to claim 1, wherein the surfactant comprises polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, or a mixture of two or more thereof.

3. The stable liquid pharmaceutical formulation according to claim 1, wherein the surfactant comprises polysorbate 80.

4. The stable liquid pharmaceutical preparation according to claim 1, having a viscosity of 0.5 cp to 10 cp after being stored at 40℃±2℃ for 1 month, or having a viscosity of 0.5 cp to 5 cp after being stored at 5℃±3℃ for 6 months.

5. A stable liquid pharmaceutical preparation according to any one of claims 1 to 4, for subcutaneous administration.

6. A stable liquid pharmaceutical preparation according to any one of claims 1 to 4, which does not undergo a reconstitution step, a dilution step, or both before use.

7. A pre-filled syringe filled with a stable liquid pharmaceutical formulation according to any one of claims 1 to 4.

8. An autoinjector comprising the pre-filled syringe of claim 7.

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