Stable liquid formulations

Abstract

The present invention relates to a liquid formulation comprising an antibody, an acetate buffer, glycine, and a surfactant wherein the liquid formulation does not comprise any one of sucrose, trehalose, mannitol, sorbitol, and NaCl, is stable even at high antibody content, and has excellent osmotic pressure and viscosity, and subcutaneous administration is feasible.

Description

[0001] This application is a divisional application of the patent application for invention with the application date of January 10, 2018, application number 201880006548.3, and invention title "Stable Liquid Formulation". Technical Field

[0002] The present invention relates to a stable liquid formulation. Background Art

[0003] Protein formulations, especially antibody formulations for intravenous injection, have been used for a rather long time. Proteins, especially antibodies, tend to form aggregates and / or dimers and become fragmented or denatured. When formulations containing them are intravenously injected, serious side effects may occur, for example, anaphylactic shock. Many attempts have been made to prevent their aggregation and fragmentation and improve their stability. For example, antibodies for intravenous injection are often lyophilized to improve their stability during storage, but such formulations must be reconstituted with a diluent before use. This reconstitution process is inconvenient and time-consuming, increasing the possibility of product contamination.

[0004] As conventional formulations containing such antibodies, various liquid formulations are known. U.S. Patent No. 8932591 discloses a stable liquid pharmaceutical formulation containing an anti-hTNFα antibody, a polyol, a surfactant, and a buffer system (citrate and phosphate). However, this formulation with a low antibody content of about 50 mg / mL has limitations in the dosage and frequency of administration and still requires improved stability.

[0005] U.S. Publication No. 2005-0260204 discloses an antibody formulation containing an antibody, histidine, a polyol, and / or NaCl. However, this liquid formulation containing NaCl as a tonicity agent may encounter problems such as precipitation and gelation and still requires improved stability.

[0006] Korean Patent Application Publication No. 10-2014-0134689 discloses a liquid formulation containing: an effective amount of an anti-TNF-α antibody or its antigen-binding portion in a buffer system (succinate), a surfactant, a tonicity agent (NaCl or KCl), and a stabilizer selected from an amino acid (arginine) and cyclodextrin. However, this liquid formulation contains NaCl or KCl as a tonicity agent, so it may cause problems such as precipitation and gelation, and due to the low antibody content of about 50 mg / mL, the dosage and frequency of administration may be limited.

[0007] Therefore, there is a need for a stable liquid formulation that can solve the problems of existing liquid formulations and contains an antibody. Summary of the Invention

[0008] SUMMARY OF THE INVENTION

[0009] Accordingly, the present invention aims to provide a stable liquid formulation.

[0010] In addition, the present invention aims to provide a stable liquid formulation that is stable even when having a high antibody content.

[0011] In addition, the present invention aims to provide a stable liquid formulation having excellent osmotic pressure and viscosity.

[0012] In addition, the present invention aims to provide a stable liquid formulation suitable for subcutaneous administration.

[0013] In a specific example of the present invention, the stable liquid formulation comprises an antibody or an antigen-binding portion thereof, an acetate buffer, glycine, and a surfactant, wherein the stable liquid formulation may not contain at least one of sugar, sugar alcohol, and metal salt.

[0014] In a specific example of the present invention, (A) the antibody may comprise a monoclonal antibody.

[0015] In a specific example of the present invention, (A) the antibody may comprise a fully human antibody.

[0016] In a specific example of the present invention, (A) the antibody may comprise an antibody that binds to TNF-α.

[0017] In a specific example of the present invention, (A) the antibody may comprise at least one of the following: infliximab, adalimumab, certolizumab pegol, and golimumab.

[0018] In a specific example of the present invention, (A) the antibody may comprise a light chain variable region comprising: a CDR1 region containing the amino acid sequence of SEQ ID NO:1, a CDR2 region containing the amino acid sequence of SEQ ID NO:2, and a CDR3 region containing the amino acid sequence of SEQ ID NO:3, and a heavy chain variable region comprising: a CDR1 region containing the amino acid sequence of SEQ ID NO:4, a CDR2 region containing the amino acid sequence of SEQ ID NO:5, and a CDR3 region containing the amino acid sequence of SEQ ID NO:6.

[0019] In a specific example of the present invention, (A) the antibody may comprise the following: a light chain variable region containing the amino acid sequence of SEQ ID NO:7 and a heavy chain variable region containing the amino acid sequence of SEQ ID NO:8.

[0020] In a specific example of the present invention, (A) the antibody may comprise the following: a light chain containing the amino acid sequence of SEQ ID NO: 9 and a heavy chain containing the amino acid sequence of SEQ ID NO: 10.

[0021] In a specific example of the present invention, (A) the antibody may have a concentration of 50 to 150 mg / mL.

[0022] In a specific example of the present invention, (B) the acetate buffer may contain acetate.

[0023] In a specific example of the present invention, the amount of acetate may be 1 to 30 mM.

[0024] In a specific example of the present invention, the stable liquid formulation may not contain at least one of the following: histidine, citrate, phosphate, malate, tartrate, and succinate.

[0025] In a specific example of the present invention, (C) the glycine may have a concentration of 100 to 300 mM.

[0026] In a specific example of the present invention, the stable liquid formulation may not contain at least one of the following: alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine.

[0027] In a specific example of the present invention, (D) the surfactant may comprise polysorbate, poloxamer, or a mixture thereof.

[0028] In a specific example of the present invention, (D) the surfactant may comprise at least one of the following: polysorbate 20, polysorbate 40, polysorbate 60, and polysorbate 80.

[0029] In a specific example of the present invention, (D) the surfactant may comprise polysorbate 80.

[0030] In a specific example of the present invention, (D) the surfactant may have a concentration of 0.01 to 1% (w / v).

[0031] In a specific example of the present invention, the pH of the stable liquid formulation may be in the range of 4.5 to 5.5.

[0032] In a specific example of the present invention, the osmotic pressure of the stable liquid formulation may be in the range of 200 to 400 mmol / kg.

[0033] In a specific example of the present invention, the stable liquid formulation may not contain a preservative, a chelating agent, or a mixture thereof.

[0034] In a specific example of the present invention, the stable liquid formulation comprises 50 to 150 mg / mL of an antibody or an antigen-binding portion thereof, an acetate buffer containing 1 to 30 mM acetate, 100 to 300 mM of glycine, and 0.01 to 1% (w / v) of a surfactant, wherein the stable liquid formulation may not contain at least one of sugar, sugar alcohol, and metal salt.

[0035] In a specific example of the present invention, the stable liquid formulation comprises (A) 50 to 150 mg / mL of an antibody or an antigen-binding portion thereof, which comprises a light chain variable region comprising a CDR1 region having the amino acid sequence of SEQ ID NO:1, a CDR2 region having the amino acid sequence of SEQ ID NO:2, and a CDR3 region having the amino acid sequence of SEQ ID NO:3; and a heavy chain variable region comprising a CDR1 region having the amino acid sequence of SEQ ID NO:4, a CDR2 region having the amino acid sequence of SEQ ID NO:5, and a CDR3 region having the amino acid sequence of SEQ ID NO:6, (B) an acetate buffer containing 1 to 30 mM of acetate, (C) 100 to 300 mM of glycine, and (D) 0.01 to 1% (w / v) of a surfactant; wherein the stable liquid formulation may not contain at least one of sugar, sugar alcohol, and metal salt.

[0036] In a specific example of the present invention, the stable liquid formulation comprises (A) 100 mg / mL of an antibody or an antigen-binding portion thereof, which comprises a light chain variable region comprising a CDR1 region having the amino acid sequence of SEQ ID NO:1, a CDR2 region having the amino acid sequence of SEQ ID NO:2, and a CDR3 region having the amino acid sequence of SEQ ID NO:3; and a heavy chain variable region comprising a CDR1 region having the amino acid sequence of SEQ ID NO:4, a CDR2 region having the amino acid sequence of SEQ ID NO:5, and a CDR3 region having the amino acid sequence of SEQ ID NO:6, (B) an acetate buffer containing 10 mM of acetate, (C) 250 mM of glycine, and (D) 0.1% (w / v) of a surfactant, wherein the stable liquid formulation may not contain at least one of sugar, sugar alcohol, and metal salt.

[0037] In a specific example of the present invention, the stable liquid formulation can be used for subcutaneous administration.

[0038] In another specific example of the present invention, a pre-filled syringe filled with the stable liquid formulation is provided.

[0039] In yet another specific example of the present invention, an auto-injector is provided, which contains the pre-filled syringe.

[0040] According to the present invention, a stable liquid formulation is stable even at high antibody contents, can exhibit excellent osmotic pressure and viscosity, and is suitable for subcutaneous administration. Detailed Description

[0041] Description of Specific Embodiments

[0042] "Stable liquid formulation"

[0043] In a specific example of the present invention, the stable liquid formulation contains an antibody or an antigen-binding portion thereof, an acetate buffer, glycine, and a surfactant, and may not contain at least one of sugars, sugar alcohols, and metal salts.

[0044] "Stable" or "stability"

[0045] As used herein, the terms "stable" or "stability" mean that the antibody according to the present invention substantially retains physical stability, chemical stability, and / or biological activity during its preparation method and / or its storage. A variety of analytical techniques for measuring antibody stability can be easily carried out in the art.

[0046] Physical stability can be evaluated by any method known in the art, including measuring the apparent attenuation of light (absorbance or optical density) of a sample. Such measurements of light attenuation are related to the turbidity of the formulation. For physical stability, the high molecular weight component content, low molecular weight component content, intact protein content, number of subvisible particles, etc. can be measured.

[0047] Chemical stability can be evaluated, for example, by detecting and quantifying the antibody in a chemically modified form. Chemical stability includes, for example, charge changes (such as those caused by deamidation or oxidation) that can be evaluated by ion exchange chromatography. For chemical stability, the charge variants (acidic or basic peaks) can be measured.

[0048] Biological activity can be evaluated by any method known in the art, including, for example, measuring antigen-binding affinity using ELISA.

[0049] As used in the present case, the term "not comprise" or "without comprising" means that the corresponding component is absolutely not included. Furthermore, the above terms mean that the corresponding component is not substantially included, i.e., included within a range that does not affect the activity of the antibody or the stability and viscosity of the liquid formulation, for example, included 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 formulation.

[0050] (A) Antibody

[0051] An antibody is an immunoglobulin molecule comprising four polypeptide chains, configured such that two heavy chains and two light chains are linked to each other by disulfide bonds. Naturally occurring antibodies with other variant structures, for example, camelid antibodies, are also included in the above definition. Each heavy chain is composed of a heavy chain variable region and a heavy chain constant region. The heavy chain constant region is composed of three regions (CH1, CH2, and CH3). Each light chain is composed of a light chain variable region and a light chain constant region. The light chain constant region is composed of one region (CL). The heavy chain variable region and the light chain variable region can be further subdivided into highly variable regions called complementarity determining regions (CDRs), which are arranged together with more conserved regions called framework regions (FRs). Each of the heavy chain variable region and the light chain variable region is composed of three CDRs and four FRs, which are arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[0052] In embodiments of the present invention, the antibody may comprise polyclonal antibodies, monoclonal antibodies, recombinant antibodies, single-chain antibodies, hybrid antibodies, chimeric antibodies, humanized antibodies, fully human antibodies, or fragments thereof. In embodiments of the present invention, the antibody may comprise monoclonal antibodies. Chimeric human-mouse monoclonal antibodies can be prepared using methods known in the art. For example, infliximab can be prepared by the method disclosed in U.S. Patent No. 6,284,471. To produce fully human antibodies to reduce the side effects of humanized or chimeric antibodies, transgenic mice and phage display technology have proven to be successful methods for their preparation. In embodiments of the present invention, the antibody may comprise fully human antibodies. Fully human monoclonal antibodies can be prepared by known methods. For example, adalimumab can be prepared by the method disclosed in U.S. Patent No. 6,090,382.

[0053] In a specific example of the present invention, the antibody may comprise an antibody that binds to TNF-α or a TNF-α epitope. The antibody that binds to TNF-α or a TNF-α epitope may include infliximab, adalimumab, certolizumab, golimumab, or a mixture thereof. In a specific example of the present invention, the antibody may comprise adalimumab.

[0054] In a specific example of the present invention, the antibody may comprise a light chain variable region comprising a CDR1 region having the amino acid sequence of SEQ ID NO:1, a CDR2 region having the amino acid sequence of SEQ ID NO:2, and a CDR3 region having the amino acid sequence of SEQ ID NO:3, and a heavy chain variable region comprising a CDR1 region having the amino acid sequence of SEQ ID NO:4, a CDR2 region having the amino acid sequence of SEQ ID NO:5, and a CDR3 region having the amino acid sequence of SEQ ID NO:6.

[0055] In a specific example of the present invention, the antibody may comprise a light chain variable region having the amino acid sequence of SEQ ID NO:7 and a heavy chain variable region having the amino acid sequence of SEQ ID NO:8.

[0056] In a specific example of the present invention, the antibody may comprise a light chain having the amino acid sequence of SEQ ID NO:9 and a heavy chain having the amino acid sequence of SEQ ID NO:10.

[0057] In the present invention, the concentration of the antibody can be 50 mg / mL or more. The concentration of the antibody can be freely adjusted within a range that has substantially no adverse effect on the stability and viscosity of the liquid formulation according to the present invention. In a specific example of the present invention, the concentration of the antibody can fall within the range of 50 to 150 mg / mL. For example, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149 or 150 mg / mL. In another specific example of the present invention, the concentration of the antibody can fall within the following ranges: 55 to 145 mg / mL, 60 to 140 mg / mL, 65 to 135 mg / mL, 70 to 130 mg / mL, 75 to 125 mg / mL, 80 to 120 mg / mL, 85 to 115 mg / mL, 90 to 110 mg / mL or 95 to 105 mg / mL. When the concentration of the antibody falls within the above ranges, due to the high content of the antibody, the degree of freedom in determining the dosage and frequency of administration can be increased, and the formulation can be better in terms of stability, viscosity and ease of preparation.

[0058] (B) Acetate buffer

[0059] The acetate buffer may contain acetate. Examples of acetate may include, but are not limited to, sodium acetate, zinc acetate, aluminum acetate, ammonium acetate and potassium acetate. The acetate buffer can be prepared by mixing acetate with acetic acid. In a specific example of the present invention, the acetate buffer may contain sodium acetate.

[0060] In a specific example of the present invention, the stable liquid formulation may not contain an additional buffer. In a specific example of the present invention, the stable liquid formulation may not contain histidine, citrate, phosphate, malate, tartrate, succinate, or a mixture thereof. When an acetate buffer is replaced or an additional buffer other than the acetate buffer is included, the stability and viscosity of the formulation may deteriorate. In particular, under UV radiation or harsh conditions, the stability of the formulation may become poor, and relatively severe pain may be caused during its subcutaneous injection.

[0061] In a specific example of the present invention, the amount of acetate in the acetate buffer can be freely adjusted within a range that has substantially no adverse effect on the stability, viscosity, and osmotic pressure of the liquid formulation according to the present invention. For example, the amount of acetate can fall within the following ranges: 0.1 to 45 mM, 1 to 30 mM, 1 to 25 mM, or 5 to 15 mM. For example, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 mM. When the amount of acetate falls within the above ranges, the stability, osmotic pressure, and viscosity of the formulation can be excellent.

[0062] Meanwhile, the amount of acetate is the acetate content of the formulation stored in a single container (vial or prefilled syringe). In a container for multiple distributions or multiple administrations, depending on the number of distributions or administrations, the amount of acetate may increase several times. Conversely, when a small container is used, the amount of acetate can be reduced to suit this.

[0063] (C) Glycine

[0064] In a specific example of the present invention, the stable liquid formulation contains glycine as an amino acid. Glycine can act as a stabilizer and can help adjust the physiological osmotic pressure. In a specific example of the present invention, the stable liquid formulation may not contain at least one of the following: alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine. When glycine is replaced or an additional amino acid other than glycine is included, the solubility may become poor, thus making it impossible to prepare the liquid formulation or destroying the stability.

[0065] In a specific example of the present invention, the amount of (C) glycine can fall within the range of 100 to 300 mM, 150 to 300 mM, or 200 to 300 mM. For example, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299 or 300 mM. When the amount of glycine falls within the above range, the osmotic pressure, stability (subvisible particles), and viscosity of the formulation can become excellent.

[0066] (D) Surfactant

[0067] Examples of surfactants can include, but are not limited to, polyoxyethylene sorbitan fatty acid esters (such as polysorbate), polyoxyethylene alkyl ethers (such as Brij), alkyl phenyl polyoxyethylene ethers (such as Triton-X), polyoxyethylene-polyoxypropylene copolymers (such as poloxamer, pluronic), and sodium dodecyl sulfate (SDS). In a specific example of the present invention, (D) the surfactant can include polysorbate, poloxamer, or a mixture thereof. In a specific example of the present invention, (D) the surfactant can include polyoxyethylene sorbitan fatty acid ester (polysorbate). In a specific example of the present invention, (D) the surfactant can include at least one of the following: polysorbate 20, polysorbate 40, polysorbate 60, and polysorbate 80. In a specific example of the present invention, (D) the surfactant can include polysorbate 20, polysorbate 80, or a mixture thereof. In a specific example of the present invention, (D) the surfactant can include polysorbate 80.

[0068] In a specific example of the present invention, (D) the concentration of the surfactant can be freely adjusted within a range that has no adverse effect on the stability and viscosity of the stable liquid formulation according to the present invention. In a specific example of the present invention, (D) the concentration of the surfactant can fall within the range of 0.001 to 5% (w / v), 0.01 to 1% (w / v), 0.02 to 1% (w / v), or 0.05 to 0.5% (w / v), for example, 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5% (w / v). When the concentration of (D) the surfactant falls within the above range, excellent stability and viscosity can be exhibited.

[0069] Unincluded or additional components

[0070] In a specific example of the present invention, the stable liquid formulation may not include at least one of sugar, sugar alcohol, and metal salt.

[0071] Sugars, such as monosaccharides, disaccharides, oligosaccharides, polysaccharides, or mixtures of two or more thereof, may not be included therein. Examples of monosaccharides may include, but are not limited to, glucose, fructose, galactose, etc. Examples of disaccharides may include, but are not limited to, sucrose, lactose, maltose, trehalose, etc. Examples of oligosaccharides may include, but are not limited to, fructooligosaccharides, galactooligosaccharides, mannanoligosaccharides, etc. Examples of polysaccharides may include, but are not limited to, starch, glycogen, cellulose, chitin, pectin, etc.

[0072] Examples of sugar alcohols may include, but are not limited to, glycerol, erythritol, threitol, arabitol, xylitol, ribitol, mannitol, sorbitol, galactitol, fucitol, iditol, inositol, volemitol, isomaltitol, maltitol, lactitol, maltotriitol, maltotetraitol, polyethanolamine, etc.

[0073] In specific examples of the present invention, sugars or sugar alcohols, such as sorbitol, mannitol, trehalose, sucrose, or mixtures of two or more thereof, may not be included.

[0074] When sugars or sugar alcohols are included, the viscosity of the liquid formulation may increase, whereby patients may experience more severe pain during subcutaneous injection.

[0075] In specific examples of the present invention, metal salts, such as NaCl, KCl, NaF, KBr, NaBr, Na2SO4, NaSCN, K2SO4, or mixtures thereof, may not be included. When such compounds are included, precipitation may occur, and the resulting formulation may have a gelatinous shape and may exhibit poor stability.

[0076] In specific examples of the present invention, chelating agents (such as EDTA) may not be included. When chelating agents are included, the oxidation rate may increase.

[0077] In specific examples of the present invention, preservatives may not be included. Examples of preservatives may include octadecyl dimethyl benzyl ammonium chloride, hexamethonium chloride, benzalkonium chloride, benzethonium chloride, phenol, butanol, benzyl alcohol, alkyl p-hydroxybenzoates, catechol, resorcinol, cyclohexanol, 3-pentanol, m-cresol, and the like. When such preservatives are included, they may not contribute to improving stability.

[0078] In a specific example of the present invention, the stable liquid formulation may further comprise additives known in the art within a range that substantially has no adverse effect on the activity of the antibody and the stability and viscosity of the formulation. For example, it may further comprise an aqueous carrier, an antioxidant, or a mixture of two or more thereof. The aqueous carrier is a pharmaceutically acceptable (safe and non-toxic after administration to the human body) carrier and can be used to prepare a liquid formulation. Examples of the aqueous carrier may include, but are not limited to, sterile water for injection (SWFI), bacteriostatic water for injection (BWFI), sterile saline solution, Ringer's solution, dextrose, and the like. Examples of the antioxidant may include, but are not limited to, ascorbic acid and the like.

[0079] pH

[0080] In a specific example of the present invention, the pH of the stable liquid formulation may be from 4.5 to 5.5, for example, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, or 5.5. In a specific example of the present invention, the pH can be adjusted using an acetate buffer. Specifically, when a predetermined amount of acetate buffer is included, the pH within the above range can be exhibited even without using an additional pH control agent. It may be difficult to achieve the pH within the above range using a buffer containing histidine, citrate, phosphate, malate, tartrate, succinate, or a mixture thereof. In the case of further comprising an acid or a base (such as sodium hydroxide) as a pH control agent, the stability of the antibody may deteriorate.

[0081] Osmotic pressure

[0082] In a specific example of the present invention, the osmotic pressure of the stable liquid formulation can be 200 to 400 mmol / kg, 250 to 350 mmol / kg, or 270 to 330 mmol / kg. For example, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370, 371, 372, 373, 374, 375, 376, 377, 378, 379, 380, 381, 382, 383, 384, 385, 386, 387, 388, 389, 390, 391, 392, 393, 394, 395, 396, 397, 398, 399 or 400 mmol / kg. When the osmotic pressure falls within the above range, the pain that may occur after subcutaneous administration can be minimized. In a specific example of the present invention, the osmotic pressure can be adjusted using an acetate buffer and glycine. Specifically, when a predetermined amount of acetate buffer and glycine are included, the osmotic pressure within the above range can be exhibited even without using an additional osmotic pressure control agent.In the case of further containing NaCl as an additional osmotic pressure control agent, precipitation may occur, and the resulting formulation may have a gelatinous form and poor stability.

[0083] Viscosity

[0084] In a specific example of the present invention, the viscosity of a stable liquid formulation measured immediately at room temperature (25°C ± 3°C) after the preparation method or measured after 6 weeks of storage at 5°C ± 3°C or 40°C ± 2°C may be 0.5 to 5.0 cp. For example, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9 or 5.0 cp. Given the above viscosity range, the pain that may occur after subcutaneous administration can be minimized, the formulation can be easily prepared and can exhibit excellent stability. When the resulting formulation is applied to a pre-filled syringe or an auto-injector, excellent plunger release force or dynamic sliding force can be generated.

[0085] In a specific example of the present invention, the term "stable" liquid formulation may refer to a liquid formulation that meets at least one of the following criteria.

[0086] Appearance analysis

[0087] - The liquid formulation, observed after storage at a temperature of 5°C ± 3°C for 6 weeks, is considered to be clear in clarity.

[0088] - The liquid formulation, observed after storage at a temperature of 5°C ± 3°C for 6 weeks, is considered to be clear in clarity.

[0089] - The liquid formulation, observed after storage at a temperature of 40°C ± 2°C for 6 weeks, is considered to be clear in clarity.

[0090] - The liquid formulation, observed after storage at a temperature of 40°C ± 2°C and a relative humidity of 75 ± 5% for 6 weeks, is considered to be clear in clarity.

[0091] Turbidity

[0092] - The liquid formulation, after storage at a temperature of 5°C ± 3°C for 6 weeks, has an absorbance of 0 to 0.0900 measured at 350 nm using a spectrophotometer.

[0093] - A liquid formulation, after being stored at a temperature of 5°C ± 3°C for 6 weeks, has an absorbance measured at 350 nm using a spectrophotometer luminance meter ranging from 0 to 0.0900.

[0094] - A liquid formulation, after being stored at a temperature of 40°C ± 2°C for 6 weeks, has an absorbance measured at 350 nm using a spectrophotometer luminance meter ranging from 0 to 0.1300.

[0095] - A liquid formulation, after being stored at a temperature of 40°C ± 2°C and a relative humidity of 75 ± 5% for 6 weeks, has an absorbance measured at 350 nm using a spectrophotometer luminance meter ranging from 0 to 0.1300.

[0096] - A liquid formulation, after being stored at a temperature of 45°C ± 2°C for 3 weeks, has an absorbance measured at 350 nm using a spectrophotometer luminance meter ranging from 0 to 0.0900.

[0097] High molecular weight component (the peak whose retention time is before the main peak (intact IgG))

[0098] - A liquid formulation, after being stored at a temperature of 5°C ± 3°C for 6 weeks, has a high molecular weight component content ranging from 0 to 0.3% as measured by size exclusion high performance liquid chromatography (SE-HPLC).

[0099] - A liquid formulation, after being stored at a temperature of 5°C ± 3°C for 6 weeks, has a high molecular weight component content ranging from 0 to 0.3% as measured by SE-HPLC.

[0100] - A liquid formulation, after being stored at a temperature of 40°C ± 2°C for 6 weeks, has a high molecular weight component content ranging from 0 to 0.9% as measured by SE-HPLC.

[0101] - A liquid formulation, after being stored at a temperature of 40°C ± 2°C and a relative humidity of 75 ± 5% for 6 weeks, has a high molecular weight component content ranging from 0 to 0.9% as measured by SE-HPLC.

[0102] - A liquid formulation, after being stored at a temperature of 45°C ± 2°C for 3 weeks, has a high molecular weight component content ranging from 0 to 1.4% as measured by SE-HPLC.

[0103] Main component content (main peak)

[0104] - A liquid formulation, after being stored at a temperature of 5°C ± 3°C for 6 weeks, has a main component content ranging from 99.7% to 100% as measured by SE-HPLC.

[0105] - A liquid formulation, after being stored at a temperature of 5°C ± 3°C for 6 weeks, has a main component content ranging from 99.7% to 100% as measured by SE-HPLC.

[0106] - Liquid formulation, after being stored at a temperature of 40°C ± 2°C for 6 weeks, the content of its main components measured by SE-HPLC is 95.0% to 100%.

[0107] - Liquid formulation, after being stored at a temperature of 40°C ± 2°C and a relative humidity of 75 ± 5% for 6 weeks, the content of its main components measured by SE-HPLC is 95.0% to 100%.

[0108] Low molecular weight components (peaks whose retention times are after the main peak (intact IgG))

[0109] - Liquid formulation, after being stored at a temperature of 5°C ± 3°C for 6 weeks, the content of its low molecular weight components measured by SE-HPLC is 0.0%.

[0110] - Liquid formulation, after being stored at a temperature of 5°C ± 3°C for 6 weeks, the content of its low molecular weight components measured by SE-HPLC is 0.0%.

[0111] - Liquid formulation, after being stored at a temperature of 40°C ± 2°C for 6 weeks, the content of its low molecular weight components measured by SE-HPLC is 0 to 4.0%.

[0112] - Liquid formulation, after being stored at a temperature of 40°C ± 2°C and a relative humidity of 75 ± 5% for 6 weeks, the content of its low molecular weight components measured by SE-HPLC is 0 to 4.0%.

[0113] Intact immunoglobulin G content

[0114] - Liquid formulation, after being stored at a temperature of 5°C ± 3°C for 6 weeks, the intact immunoglobulin G content (intact IgG%) measured by non-reducing capillary electrophoresis - sodium dodecyl sulfate (NR CE-SDS) is 98.0% to 100%.

[0115] - Liquid formulation, after being stored at a temperature of 5°C ± 3°C for 6 weeks, the intact immunoglobulin G content (intact IgG%) measured by NR CE-SDS is 98.0% to 100%.

[0116] - Liquid formulation, after being stored at a temperature of 40°C ± 2°C for 6 weeks, the intact immunoglobulin G content (intact IgG%) measured by NR CE-SDS is 93.1% to 100%.

[0117] - Liquid formulation, after being stored at a temperature of 40°C ± 2°C and a relative humidity of 75 ± 5% for 6 weeks, the intact immunoglobulin G content (intact IgG%) measured by NR CE-SDS is 93.1% to 100%.

[0118] Number of sub-visible particles

[0119] – A liquid formulation having 0 to 10,000 sub-visible particles (1.00 μm ≤, < 100.00 μm) as measured by microflow imaging (MFI) after storage at a temperature of 40°C ± 2°C for 6 weeks.

[0120] - A liquid formulation having 0 to 10,000 sub-visible particles (1.00 μm ≤, < 100.00 μm) as measured by MFI after storage at a temperature of 40°C ± 2°C and a relative humidity of 75 ± 5% for 6 weeks.

[0121] - A liquid formulation having 0 to 5,000 sub-visible particles (1.00 μm ≤, < 100.00 μm) as measured by MFI after storage at a temperature of 45°C ± 2°C for 3 weeks.

[0122] Method for preparing a stable liquid formulation

[0123] The stable liquid formulation according to the present invention can be prepared using any known method, and its preparation is not limited to a specific method. For example, the liquid formulation of the present invention can be prepared by adding an acetate buffer to adjust the pH of a solution containing glycine and a surfactant, and then adding an antibody to the resulting mixed solution.

[0124] In a specific example of the present invention, the lyophilization method may or may not be carried out when preparing the liquid formulation.

[0125] When the lyophilization method is not carried out, for example, the liquid formulation of the present invention can be prepared and immediately placed in a container after processing such as aseptic processing or similar processing.

[0126] When the lyophilization method is carried out, for example, the liquid formulation of the present invention can be prepared and lyophilized, or the liquid formulation of the present invention can be prepared, lyophilized and stored, and then any components removed or modified by lyophilization and / or storage are supplemented or replaced, thereby obtaining the liquid formulation of the present invention. Furthermore, only components other than those that can be removed or modified by lyophilization and / or storage in the liquid formulation of the present invention can be lyophilized, or lyophilized and stored, and then the excluded components can be added thereto, thereby obtaining the liquid formulation of the present invention.

[0127] Method for using a stable liquid formulation

[0128] The stable liquid formulation according to the present invention can be used to treat diseases targeted by the corresponding antibody, for example, diseases harmful to TNF-α activity. Examples of diseases harmful to TNF-α activity may include, but are not limited to, sepsis, autoimmune diseases, infectious diseases, transplantation, malignant cancers, lung diseases, intestinal diseases, heart diseases, and the like.

[0129] In a specific example of the present invention, diseases in which TNF-α activity is harmful may be selected from rheumatoid arthritis, ankylosing spondylitis, ulcerative colitis, adult Crohn's disease, pediatric Crohn's disease, psoriasis, and psoriatic arthritis.

[0130] The stable liquid formulation of the present invention can be used once or several times, or for subcutaneous self-administration.

[0131] The component concentrations of the liquid formulation containing the antibody are as described above, and the total volume of the liquid formulation can fall within the range of 0.2 to 10.0 mL. For example, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9 or 10.0 mL.

[0132] The dosage and timing of administration of the liquid formulation of the present invention may depend on the type of disease, the severity and progression of the disease, the health of the patient and the treatment regimen, and the judgment of the treating physician, and are not limited to specific values. For example, based on the mass of the antibody, a single product or several products containing the liquid formulation may be administered in an amount of 0.1 to 10 mg / kg (for example, 5 to 500 mg of antibody in the case of a patient with a body weight of 50 kg), and then the same or different amounts may be administered weekly, every other week, every 3 weeks, monthly, every 2 months, or every 3 months. Furthermore, the dosage and timing of administration of the liquid formulation may be determined with reference to the approved drug containing the antibody, for example, the Humira label.

[0133] Treatment methods and stabilization methods

[0134] The present invention provides a method for treating a disease targeted by a corresponding antibody (for example, a disease in which the activity of TNF-α is harmful), the method comprising administering to a patient suffering from a disease targeted by a corresponding antibody, for example, a disease in which the activity of TNF-α is harmful, a stable liquid formulation comprising (A) an antibody or an antigen-binding portion thereof, (B) an acetate buffer, (C) glycine, and (D) a surfactant, and not comprising at least one of sugars, sugar alcohols, and metal salts.

[0135] Furthermore, the present invention provides a method for stabilizing an antibody in a liquid formulation, the method comprising preparing a stable liquid formulation comprising (A) an antibody or an antigen-binding portion thereof, (B) an acetate buffer, (C) glycine, and (D) a surfactant, and not comprising at least one of sugars, sugar alcohols, and metal salts.

[0136] In a specific example of the treatment method or the stabilization method, (A) the antibody may comprise a monoclonal antibody.

[0137] In a specific example of the treatment method or the stabilization method, (A) the antibody may comprise a fully human antibody.

[0138] In a specific example of the treatment method or the stabilization method, (A) the antibody may comprise an antibody that binds to TNF-α.

[0139] In a specific example of the treatment method or the stabilization method, (A) the antibody may comprise at least one of the following: infliximab, adalimumab, certolizumab, and golimumab.

[0140] In a specific example of the treatment method or the stabilization method, (A) the antibody may comprise a light chain variable region comprising: a CDR1 region containing the amino acid sequence of SEQ ID NO:1, a CDR2 region containing the amino acid sequence of SEQ ID NO:2, and a CDR3 region containing the amino acid sequence of SEQ ID NO:3; and a heavy chain variable region comprising: a CDR1 region containing the amino acid sequence of SEQ ID NO:4, a CDR2 region containing the amino acid sequence of SEQ ID NO:5, and a CDR3 region containing the amino acid sequence of SEQ ID NO:6.

[0141] In a specific example of the treatment method or the stabilization method, (A) the antibody may comprise a light chain variable region containing the amino acid sequence of SEQ ID NO:7, and a heavy chain variable region containing the amino acid sequence of SEQ ID NO:8.

[0142] In a specific example of the treatment method or the stabilization method, (A) the antibody may comprise a light chain containing the amino acid sequence of SEQ ID NO:9, and a heavy chain containing the amino acid sequence of SEQ ID NO:10.

[0143] In a specific example of a treatment method or a stabilization method, (A) the antibody may have a concentration of 50 to 150 mg / mL.

[0144] In a specific example of a treatment method or a stabilization method, (B) the acetate buffer may contain acetate.

[0145] In a specific example of a treatment method or a stabilization method, the amount of acetate may be 1 to 30 mM.

[0146] In a specific example of a treatment method or a stabilization method, the stable liquid formulation may not contain at least one of the following: histidine, citrate, phosphate, malate, tartrate, and succinate.

[0147] In a specific example of a treatment method or a stabilization method, (C) glycine may have a concentration of 100 to 300 mM.

[0148] In a specific example of a treatment method or a stabilization method, the stable liquid formulation may not contain at least one of the following: alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine.

[0149] In a specific example of a treatment method or a stabilization method, (D) the surfactant may contain polysorbate, poloxamer, or a mixture thereof.

[0150] In a specific example of a treatment method or a stabilization method, (D) the surfactant may contain at least one of the following: polysorbate 20, polysorbate 40, polysorbate 60, and polysorbate 80.

[0151] In a specific example of a treatment method or a stabilization method, (D) the surfactant may contain polysorbate 80.

[0152] In a specific example of a treatment method or a stabilization method, (D) the surfactant may have a concentration of 0.01 to 1% (w / v).

[0153] In a specific example of a treatment method or a stabilization method, the stable liquid formulation may have a pH in the range of 4.5 to 5.5.

[0154] In a specific example of a treatment method or a stabilization method, the stable liquid formulation may have an osmotic pressure in the range of 200 to 400 mmol / kg.

[0155] In a specific example of a treatment method or a stabilization method, the stable liquid formulation may not contain a preservative, a chelating agent, or a mixture thereof.

[0156] In a specific example of a treatment method or a stabilization method, the stable liquid formulation may further contain an aqueous carrier, an antioxidant, or a mixture of two or more of them.

[0157] In a specific example of a treatment method or a stabilization method, the stable liquid formulation may have a viscosity in the range of 0.5 to 5.0 cp.

[0158] In a specific example of a treatment method or a stabilization method, the stable liquid formulation may be used for subcutaneous administration.

[0159] Product

[0160] The present invention provides a product comprising the stable liquid formulation and a container containing the stable pharmaceutical formulation in a closed state.

[0161] The stable liquid formulation is as described above.

[0162] In a specific example of the present invention, the container may be formed of glass, polymer (plastic), metal, or the like, but is not limited thereto. In a specific example of the present invention, the container is a bottle, a vial, a syringe, for example, a pre-filled or pre-fillable syringe, or a tube, but is not limited thereto. In a specific example of the present invention, the container may be a vial made of glass or polymer, or a pre-filled syringe made of glass or polymer. In a specific example of the present invention, a pre-filled syringe filled with the stable liquid formulation is provided.

[0163] In a specific example of the present invention, the inner surface of the pre-filled syringe may be coated with silicone oil. In this case, excellent plunger release force or dynamic sliding force may occur. In a specific example of the present invention, the inner surface of the pre-filled syringe may not be coated with silicone oil. In this case, the stability of the formulation may be excellent. The container may be a single-dose or multi-dose container.

[0164] In a specific example of the present invention, the product may be an auto-injector, and the auto-injector may contain a pre-filled syringe filled with the stable liquid formulation. The auto-injector may include, for example, a cylindrical housing containing the pre-filled syringe and an actuator (such as a spring) that starts administration by applying pressure to the stopper of the pre-filled syringe, and may be composed of glass, polymer (plastic), or metal. As the auto-injector, any one of known products may be used, or a product containing the pre-filled syringe may be customized.

[0165] In a specific example of the present invention, the product may further include instructions for any one or both of a method of using the stable liquid formulation and a method of storing the stable liquid formulation. In a specific example of the present invention, the method of use includes treating a disease targeted by the corresponding antibody, for example, a disease in which the activity of TNF-α is harmful, and may include the route of administration, dosage, and timing.

[0166] In a specific example of the present invention, the product may include other tools necessary from a commercial purpose and the perspective of the user, for example, needles, syringes, and the like.

[0167] The present invention will be better understood through the following examples, which are presented for illustration only and should not be construed as limiting the scope of the present invention.

[0168] Examples

[0169] Regarding the antibodies used in Examples 1 and 2 and Comparative Examples 1 to 5 and 7 to 12, adalimumab manufactured by Celltrion was used. Regarding the antibody used in Comparative Example 6, adalimumab manufactured by AbbVie was used.

[0170] In order to measure the physical and chemical stability of the liquid formulations of the examples, the following methods were used.

[0171] - Appearance analysis

[0172] The clarity of the formulation was observed.

[0173] - Turbidity

[0174] The absorbance was measured at 350 nm using a UV-Vis spectrophotometer.

[0175] - Content of main components

[0176] The content of main components (main peak; %) was measured using size exclusion high performance liquid chromatography (SE-HPLC).

[0177] - Content of high molecular weight components

[0178] The content of high molecular weight components (before the peak; %) was measured using SE-HPLC.

[0179] - Content of low molecular weight components

[0180] The content of low molecular weight components (after the peak; %) was measured using SE-HPLC.

[0181] - Content of intact immunoglobulin G (intact IgG %)

[0182] The intact IgG content (%) was measured using non-reducing capillary electrophoresis-sodium dodecyl sulfate (NR CE-SDS).

[0183] -Number of sub-visible particles

[0184] The number of subvisible particles (1.00 μm ≤, <100.00 μm) was measured using microflow imaging (MFI).

[0185] -Osmotic pressure

[0186] Osmolality (mmol / kg) was measured using an osmometer (VAPRO 5520).

[0187] - Stickiness

[0188] The viscosity of a 500 μL syringe was measured at 25°C ± 0.1°C using a microcapillary rheometer (apparent shear rate: 103 to 105 s-1) equipped with a flow cell (B05 sensor type, 50 μm cell depth).

[0189] Examples 1 and 2 and Comparative Examples 1 to 12

[0190] In the liquid formulations of Examples 1 and 2 and Comparative Examples 1 to 12, each buffer was prepared to adjust to the corresponding pH, an amino acid, a metal salt, a sugar, or a sugar alcohol was added, an antibody was added, and a surfactant was further added, thereby producing the samples listed in Table 1 below. The specific amounts of the individual components are shown in Table 1 below. The total volume was 3 mL.

[0191] [Table 1]

[0192]

[0193]

[0194] The liquid formulations of Examples 1 and 2 and Comparative Examples 1 to 7 were stored at a temperature of 5±3° C., a humidity of 40±2° C., and a relative humidity of 75±5% for 6 weeks.

[0195] Furthermore, the liquid formulations of Example 1 and Comparative Examples 8 to 12 were stored at a temperature of 45±2° C. for 3 weeks.

[0196] Appearance analysis

[0197] [Table 2]

[0198]

[0199] Referring to Table 2, from the time point of preparation, the liquid formulations of Examples 1 and 2 were relatively clear compared to Comparative Examples 1 to 5, and no changes in appearance over time under respective storage conditions were observed.

[0200] Turbidity

[0201] [Table 3]

[0202]

[0203]

[0204] Referring to Table 3, Examples 1 and 2 - containing acetate buffer and glycine - are the best in terms of turbidity. Even after 6 weeks at 40°C, the absorbance of Examples 1 and 2 is still lower than that of Comparative Examples 1 to 6.

[0205] [Table 4]

[0206]

[0207] Referring to Table 4, Example 1 - containing 10 mM acetate buffer and 250 mM glycine - is the best in terms of turbidity. In particular, even after 3 weeks at 45°C, the absorbance of Example 1 is 0.1800 or less. Thus, it is still lower than that of Comparative Examples 8 to 12.

[0208] High - molecular - weight component content

[0209] [Table 5]

[0210]

[0211] Referring to Table 5, the high - molecular - weight component content of Example 1 is the lowest under all conditions. In particular, after 6 weeks at 40°C, the high - molecular - weight component content of Example 1 is less than 1.0%.

[0212] [Table 6]

[0213]

[0214]

[0215] Referring to Table 6, after 3 weeks at 45°C, the high - molecular - weight component content of Example 1 is less than 1.5%.

[0216] Main component content

[0217] [Table 7]

[0218]

[0219] Referring to Table 7, after 6 weeks at 40°C, the monomer content of Example 1 is 95.0% or more, which is higher than that of Comparative Examples 1 to 6.

[0220] [Table 8]

[0221]

[0222] Referring to Table 8, after 3 weeks at 45°C, the monomer content of Example 1 was 95.0% or more, which was higher than that of Comparative Examples 11 and 12.

[0223] Low molecular weight component content

[0224] [Table 9]

[0225]

[0226]

[0227] Referring to Table 9, after 6 weeks at 40°C, the low molecular weight component content of Example 1 was less than 4%, which was lower than that of Comparative Examples 1 to 6.

[0228] Content of intact immunoglobulin G (intact IgG%)

[0229] [Table 10]

[0230]

[0231] Referring to Table 10, after 6 weeks at 40°C, the intact IgG% of Example 1 was 93.10% or more, which was higher than that of Comparative Examples 1 to 6.

[0232] Number of sub-visible particles (1.00 μm ≤, < 100.00 μm)

[0233] [Table 11]

[0234]

[0235] Referring to Table 11, in Example 1 or 2, where 250 or 280 mM glycine was used, after 6 weeks at 40°C, the number of sub-visible particles was 10,000 or less. However, in Comparative Example 6 in the form of Humira formulation, after 6 weeks at 40°C, the number of sub-visible particles was 50,000 or more. Furthermore, in Comparative Example 7 using 310 mM glycine, after 6 weeks at 40°C, the number of sub-visible particles was 35,000 or more.

[0236] [Table 12]

[0237]

[0238]

[0239] Referring to Table 12, in Example 1 using 250 mM glycine, after 3 weeks at 45°C, the number of sub-visible particles was 5000 or less. However, in Comparative Examples 8 and 12, after 3 weeks at 45°C, the number of sub-visible particles was 4,000,000 or more.

[0240] Osmotic pressure

[0241] [Table 13]

[0242]

[0243] Referring to Table 13, the osmotic pressure of Comparative Example 10 was less than 200 mmol / kg, which was lower than that of Example 1 and Comparative Examples 8, 9, 11, and 12.

[0244] Viscosity

[0245] [Table 14]

[0246]

[0247] Referring to Table 14, after 3 weeks at 45°C, the viscosity of Example 1 was measured to be less than 3.0.

[0248]

[0249]

[0250]

Claims

1. A liquid formulation, comprising: (A) an antibody; (B) an acetate buffer; (C) glycine; and (D) a surfactant, wherein the liquid formulation does not contain any of sucrose, trehalose, mannitol, sorbitol, and NaCl, wherein after storage at a temperature of 40°C ± 2°C for 6 weeks, the number of sub-visible particles in the liquid formulation measured by microfluidic imaging (MFI) is 0 to 10,000.

2. The liquid formulation according to claim 1, wherein after storage at a temperature of 45°C ± 2°C for 3 weeks, the high molecular weight component content in the liquid formulation measured by SE-HPLC is 0 to 1.4%.

3. The liquid formulation according to claim 1, wherein after storage at a temperature of 40°C ± 2°C and a relative humidity of 75 ± 5% for 6 weeks, the absorbance of the liquid formulation measured at 350 nm using a spectrophotometer is 0 to 0.1300; or After storing at a temperature of 40°C ± 2°C for 6 weeks, the content of the high molecular weight component in the liquid formulation was measured by SE-HPLC to be 0 to 0.9%; or after storage at a temperature of 40°C ± 2°C for 6 weeks, the main component content in the liquid formulation measured by SE-HPLC is 95.0% to 100%; or after storage at a temperature of 40°C ± 2°C and a relative humidity of 75 ± 5% for 6 weeks, the low molecular weight component content in the liquid formulation measured by SE-HPLC is 0 to 4.0%; or after storage at a temperature of 40°C ± 2°C for 6 weeks, the intact immunoglobulin G content (intact IgG%) in the liquid formulation measured by NR CE-SDS is 93.1% to 100%.

4. The liquid formulation according to claim 1, wherein (A) the antibody comprises at least one of the following: infliximab, adalimumab, certolizumab, and golimumab.

5. The liquid formulation according to claim 1, wherein (A) the antibody comprises: a light chain variable region comprising a CDR1 region having the amino acid sequence shown in SEQ ID NO:1, a CDR2 region having the amino acid sequence shown in SEQ ID NO:2, and a CDR3 region having the amino acid sequence shown in SEQ ID NO:3; and a heavy chain variable region comprising a CDR1 region having the amino acid sequence shown in SEQ ID NO:4, a CDR2 region having the amino acid sequence shown in SEQ ID NO:5, and a CDR3 region represented by the amino acid sequence of SEQ ID NO:

6.

6. The liquid formulation according to claim 1, wherein (A) the antibody has a concentration of 50 to 150 mg / mL.

7. The liquid formulation according to claim 1, wherein (B) the acetate buffer is 1 to 30 mM.

8. The liquid formulation according to claim 1, wherein the liquid formulation does not contain at least one of the following: histidine, citrate, phosphate, malate, tartrate, and succinate.

9. The liquid formulation according to claim 1, wherein (C) the glycine has a concentration of 100 to 300 mM.

10. The liquid formulation according to claim 1, wherein the liquid formulation does not contain at least one of the following: alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine.

11. The liquid formulation according to claim 1, wherein (D) the surfactant comprises polysorbate, poloxamer, or a mixture thereof.

12. The liquid formulation according to claim 1, wherein (D) the surfactant comprises at least one of the following: polysorbate 20, polysorbate 40, polysorbate 60, and polysorbate 80.

13. The liquid formulation according to claim 1, wherein (D) the surfactant has a concentration of 0.01 to 1% (w / v).

14. The liquid formulation according to claim 1, which has a pH in the range of 4.5 to 5.

5.

15. The liquid formulation according to claim 1, which has an osmotic pressure in the range of 200 to 400 mmol / kg.

16. The liquid formulation according to claim 1, wherein the liquid formulation does not contain a preservative, a chelating agent, or a mixture thereof.

17. The liquid formulation according to claim 1, comprising: (A) 50 to 150 mg / mL of an antibody; (B) an acetate buffer containing 1 to 30 mM of acetate; (C) 100 to 300 mM of glycine; and (D) 0.01 to 1% (w / v) of a surfactant, wherein the liquid formulation does not contain any of sucrose, trehalose, mannitol, sorbitol, and NaCl.

18. The liquid formulation according to claim 1, which comprises: (A) 50 to 150 mg / mL of an antibody, which comprises a light chain variable region and a heavy chain variable region; the light chain variable region comprises a CDR1 region having the amino acid sequence shown in SEQ ID NO:1, a CDR2 region having the amino acid sequence shown in SEQ ID NO:2, and a CDR3 region having the amino acid sequence shown in SEQ ID NO:3; the heavy chain variable region comprises a CDR1 region having the amino acid sequence shown in SEQ ID NO:4, a CDR2 region having the amino acid sequence shown in SEQ ID NO:5, and a CDR3 region having the amino acid sequence shown in SEQ ID NO:6; (B) an acetate buffer containing 1 to 30 mM of acetate; (C) 100 to 300 mM of glycine; and (D) 0.01 to 1% (w / v) of a surfactant, wherein the liquid formulation does not contain any of sucrose, trehalose, mannitol, sorbitol, and NaCl.

19. The liquid formulation according to claim 1, comprising: (A) An antibody at 100 mg / mL, which comprises a light chain variable region and a heavy chain variable region; the light chain variable region comprises: a CDR1 region with the amino acid sequence shown in SEQ ID NO:1, a CDR2 region with the amino acid sequence shown in SEQ ID NO:2, and a CDR3 region with the amino acid sequence shown in SEQ ID NO:3; the heavy chain variable region comprises: a CDR1 region with the amino acid sequence shown in SEQ ID NO:4, a CDR2 region with the amino acid sequence shown in SEQ ID NO:5, and a CDR3 region with the amino acid sequence shown in SEQ ID NO:6; (B) An acetate buffer, which comprises 10 mM acetate; (C) 250 mM glycine; and (D) A surfactant at 0.1% (w / v), wherein the liquid formulation does not contain any one of sucrose, trehalose, mannitol, sorbitol, and NaCl.

20. The liquid formulation according to claim 1, which is for subcutaneous administration.

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