Pharmaceutical composition containing human anti-IL-33 monoclonal antibody

Abstract

The object of the present invention is to provide a pharmaceutical composition containing human anti-IL-33 monoclonal antibodies suitable for administration to a subject. The present invention provides a pharmaceutical composition containing human anti-IL-33 monoclonal antibodies as an active ingredient, which substantially does not contain sodium chloride, or contains less than 30 mM sodium chloride. In addition, the present invention provides a lyophilized agent of the aforementioned pharmaceutical composition.

Description

Technical Field

[0001] The present invention relates to a pharmaceutical composition containing a human anti-IL-33 monoclonal antibody (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02), and in particular to a pharmaceutical composition containing the antibody with suppressed white turbidity and improved storage stability such as pH. Background Art

[0002] In recent years, various pharmaceutical compositions containing antibodies have been developed for practical use, and many pharmaceutical compositions containing antibodies are used as pharmaceutical compositions for intravenous injection. On the other hand, due to the needs of medical sites, the demand for developing pharmaceutical compositions containing antibodies as pharmaceutical compositions for subcutaneous injection that can be self-injected is gradually increasing.

[0003] When designing a pharmaceutical composition containing an antibody for subcutaneous injection, the amount of antibody administered per time is relatively large (about 100 to 200 mg). On the other hand, subcutaneous injection generally has a limit on the amount of injection solution, so it is necessary to make the antibody in the administration solution have a high concentration. Therefore, pharmaceutical compositions containing high-concentration antibodies that utilize freeze-drying concentration technology (i.e., preparing a pharmaceutical composition containing high-concentration antibodies by re-dissolving a freeze-dried pharmaceutical composition with water less than the volume before freeze-drying) are often used.

[0004] IgG antibodies, which are one of the antibody subtypes and are commonly used in medicine, are high molecular weight glycoproteins of about 150 kDa. Since the structures of the variable regions of antibodies (amino acid sequences, sugar chain structures) are very diverse, they exhibit different physical properties depending on the molecular type of the antibody.

[0005] Antibodies may denature due to physical stimulation such as heating and vibration, interaction with compounds such as surfactants, redox agents, and sugars, or long-term storage, resulting in aggregation, decomposition, and chemical reactions. If antibodies denature, their binding properties to antigens and Fc receptors change, so there is a concern that the function and effect of the antibodies will be weakened, or that the aggregated antibodies will cause inflammation. The ease of denaturation such as aggregation varies depending on the molecular type of the antibody.

[0006] Antibodies contain charged amino acids, highly polar amino acids, and sugar chains, so they may interact with ions in the antibody solution to produce an unbalanced distribution of ions, causing changes in the pH of the antibody solution (pH drift) (Donnan effect). Changes in the pH of the antibody solution may sometimes affect the storage stability of the antibody, and the strength of the Donnan effect varies depending on the molecular type of the antibody.

[0007] High-concentration antibody solutions tend to form solutions with high viscosity due to the properties of proteins as giant molecules and intermolecular interactions. If the viscosity of the antibody solution increases, it becomes difficult to administer the antibody. The increase in the viscosity of the antibody solution caused by increasing the concentration of the antibody varies depending on the molecular type of the antibody.

[0008] Furthermore, when an antibody solution is stored for a long period of time, deterioration phenomena represented by changes in pH and the formation of insoluble and / or soluble aggregates become issues, but this also varies depending on the molecular type of the antibody.

[0009] Generally, various efforts have been made to prepare pharmaceutical compositions containing antibodies in order to minimize the loss of active ingredients even after long-term storage and stabilize the compositions, and the antibodies as active ingredients and various additives such as buffers are dissolved and prepared as pharmaceutical compositions. However, in particular, in pharmaceutical compositions containing high concentrations of antibodies, the technology for preventing aggregation, turbidity, viscosity increase, and pH drift of the antibodies is still insufficient.

[0010] The inventors of the present application have obtained a plurality of human anti-IL-33 monoclonal antibodies that bind to IL-33 (Patent Document 1: International Publication No. 2015 / 099175), but there is still a need to develop pharmaceutical compositions containing these antibodies that are suitable for administration.

[0011] Prior art literature

[0012] Patent Literature

[0013] Patent Document 1: International Publication No. 2015 / 099175 Summary of the invention

[0014] Problems to be solved by the invention

[0015] An object of the present invention is to provide a pharmaceutical composition containing a human anti-IL-33 monoclonal antibody suitable for administration to a subject.

[0016] Means for solving problems

[0017] The inventors of the present application have found that when human anti-IL-33 monoclonal antibodies are formulated, white turbidity occurs, and that the cause of the white turbidity is the formation of aggregates. In addition, it has been found that the white turbidity depends on the NaCl concentration and pH. Based on the above findings, the present invention has been completed.

[0018] That is, the present invention provides the following.

[0019] [1] A pharmaceutical composition comprising a human anti-IL-33 monoclonal antibody as an active ingredient, wherein:

[0020] The combination of amino acid sequences of the heavy chain complementarity determining region 1 (H1), the heavy chain complementarity determining region 2 (H2), the heavy chain complementarity determining region 3 (H3), the light chain complementarity determining region 1 (L1), the light chain complementarity determining region 2 (L2), and the light chain complementarity determining region 3 (L3) of the human anti-IL-33 monoclonal antibody is any one of C1 to C5 in Table 1,

[0021] The pharmaceutical composition contains substantially no sodium chloride or contains less than 30 mM sodium chloride.

[0022] [Table 1]

[0023] The sequence numbers below Table 1 indicate the sequence numbers of the sequence listing.

[0024] H1 H2 H3 L1 L2 L3 C1 Sequence Number 11 Sequence Number 12 Sequence Number 13 Sequence Number 14 Sequence number 15 Sequence Number 16 C2 Sequence Number 17 Sequence Number 18 Sequence Number 19 Sequence number 20 Sequence Number 21 Sequence Number 22 C3 Sequence Number 17 Sequence Number 23 Sequence Number 24 Sequence number 25 Sequence Number 26 Sequence Number 27 C4 Sequence number 28 Sequence Number 29 Sequence number 30 Sequence number 25 Sequence Number 31 Sequence number 32 C5 Sequence Number 17 Sequence Number 33 Sequence Number 34 Sequence Number 35 Sequence Number 36 Serial Number 37

[0025] [2] The pharmaceutical composition according to [1], wherein the combination of amino acid sequences of the heavy chain variable region and the light chain variable region of the human anti-IL-33 monoclonal antibody is any one of V1 to V5 in Table 2.

[0026] [Table 2]

[0027] The sequence numbers below Table 2 represent the sequence numbers of the sequence listing.

[0028]

[0029]

[0030] [3] The pharmaceutical composition according to [1] or [2], wherein the human anti-IL-33 monoclonal antibody is A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02.

[0031] [4] The pharmaceutical composition according to any one of [1] to [3], wherein the sodium chloride concentration is 10 mM or less.

[0032] [5] The pharmaceutical composition according to any one of [1] to [4], which does not substantially contain sodium chloride.

[0033] [6] The pharmaceutical composition according to any one of [1] to [5], wherein the pH of the pharmaceutical composition is adjusted to be greater than 4 and less than 8.

[0034] [7] The pharmaceutical composition according to any one of [1] to [6], wherein the pH of the pharmaceutical composition is adjusted to 5 or more and 7 or less.

[0035] [8] The pharmaceutical composition according to any one of [1] to [7], wherein the pH is adjusted using a buffer such as acetate, histidine or phosphate.

[0036] [9] The pharmaceutical composition according to any one of [1] to [8], wherein the pH is adjusted with histidine.

[0037]

[10] The pharmaceutical composition according to any one of [1] to [9], wherein the concentration of the active ingredient is less than 175 mg / ml.

[0038]

[11] The pharmaceutical composition according to any one of [1] to

[10] , wherein the concentration of the active ingredient is 150 mg / ml or less.

[0039]

[12] The pharmaceutical composition according to any one of [1] to

[11] , comprising at least one polyol.

[0040] [12-1] The pharmaceutical composition according to

[12] , wherein the polyol is a sugar selected from the group consisting of disaccharides and sugar alcohols.

[0041]

[13] The pharmaceutical composition according to

[12] or [12-1], wherein the polyol is 3 to 5% (w / v) sorbitol.

[0042]

[14] The pharmaceutical composition according to any one of [1] to

[13] , which contains a surfactant.

[0043]

[15] The pharmaceutical composition according to

[14] , wherein the surfactant is a nonionic surfactant.

[0044] [15-1] The pharmaceutical composition according to

[15] , wherein the surfactant is polysorbate 20, polysorbate 80 or poloxamer 188.

[0045]

[16] The pharmaceutical composition as described in any one of [1] to [15-1], comprising 10 mM histidine, 4% (w / v) sorbitol, 0.02% (w / v) polysorbate 80, and 150 mg / ml of the active ingredient, and the pH is adjusted to 5.5 to 6.5.

[0046]

[17] The pharmaceutical composition according to

[16] , which is for subcutaneous administration.

[0047]

[18] The pharmaceutical composition as described in any one of [1] to [15-1], comprising 10 mM histidine, 3.6% (w / v) sorbitol, 0.02% (w / v) polysorbate 80, and 10 mg / ml of the active ingredient, and the pH is adjusted to 5.5 to 6.5.

[0048]

[19] The pharmaceutical composition according to

[18] , which is for intravenous administration.

[0049]

[20] The pharmaceutical composition according to any one of [1] to

[19] , wherein the active ingredient is A10-1C04.

[0050]

[21] A freeze-dried preparation of the pharmaceutical composition according to any one of [1] to

[20] .

[0051]

[22] A method for treating or preventing an IL-33-related disease, characterized in that:

[0052] The method comprises administering a pharmaceutical composition comprising a human anti-IL-33 monoclonal antibody as an active ingredient to a subject in need thereof,

[0053] The combination of amino acid sequences of the heavy chain complementarity determining region 1 (H1), the heavy chain complementarity determining region 2 (H2), the heavy chain complementarity determining region 3 (H3), the light chain complementarity determining region 1 (L1), the light chain complementarity determining region 2 (L2), and the light chain complementarity determining region 3 (L3) of the human anti-IL-33 monoclonal antibody is any one of C1 to C5 in Table 1 above,

[0054] The pharmaceutical composition contains substantially no sodium chloride or contains less than 30 mM sodium chloride.

[0055]

[23] The method of

[22] , wherein the combination of amino acid sequences of the heavy chain variable region and the light chain variable region of the human anti-IL-33 monoclonal antibody is any one of V1 to V5 in Table 2 above.

[0056]

[24] The method of

[22] or

[23] , wherein the human anti-IL-33 monoclonal antibody is A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02.

[0057]

[25] The method according to any one of

[22] to

[24] , wherein the sodium chloride concentration of the pharmaceutical composition is 10 mM or less.

[0058]

[26] The method according to any one of

[22] to

[25] , wherein the pharmaceutical composition contains substantially no sodium chloride.

[0059]

[27] The method according to any one of

[22] to

[26] , wherein the pH of the pharmaceutical composition is adjusted to be greater than 4 and less than 8.

[0060]

[28] The method according to any one of

[22] to

[27] , wherein the pH of the pharmaceutical composition is adjusted to 5 or more and 7 or less.

[0061]

[29] The method according to any one of

[22] to

[28] , wherein the pH of the pharmaceutical composition is adjusted using an acetate, histidine or phosphate buffer.

[0062]

[30] The method according to any one of

[22] to

[29] , wherein the pH of the pharmaceutical composition is adjusted using histidine.

[0063]

[31] The method according to any one of

[22] to

[30] , wherein the concentration of the active ingredient is less than 175 mg / ml.

[0064]

[32] The method according to any one of

[22] to

[31] , wherein the concentration of the active ingredient is 150 mg / ml or less.

[0065]

[33] The method according to any one of

[22] to

[32] , wherein the pharmaceutical composition contains at least one selected from the group consisting of sorbitol, sucrose, trehalose, and mannitol.

[0066]

[34] The method according to any one of

[22] to

[33] , wherein the pharmaceutical composition contains 3 to 5% (w / v) sorbitol.

[0067]

[35] The method according to any one of

[22] to

[34] , wherein the pharmaceutical composition contains a surfactant.

[0068]

[36] The method as described in

[35] , wherein the surfactant is polysorbate 20, polysorbate 80 or poloxamer 188.

[0069]

[37] The method as described in any one of

[22] to

[36] , wherein the pharmaceutical composition contains 10 mM histidine, 4% (w / v) sorbitol, 0.02% (w / v) polysorbate 80, and 150 mg / ml of the active ingredient, and the pH is adjusted to 5.5 to 6.5.

[0070]

[38] The method of

[37] , wherein the administration is subcutaneous administration.

[0071]

[39] The method as described in any one of

[22] to

[36] , wherein the pharmaceutical composition contains 10 mM histidine, 3.6% (w / v) sorbitol, 0.02% (w / v) polysorbate 80, and 10 mg / ml of the active ingredient, and the pH is adjusted to 5.5 to 6.5.

[0072]

[40] The method of

[39] , wherein the administration is intravenous administration.

[0073]

[41] The method according to any one of

[22] to

[40] , wherein the active ingredient is A10-1C04.

[0074]

[42] A pharmaceutical composition for use in the treatment or prevention of an IL-33-related disease, characterized in that:

[0075] Contains human anti-IL-33 monoclonal antibody as an active ingredient,

[0076] The combination of amino acid sequences of the heavy chain complementarity determining region 1 (H1), the heavy chain complementarity determining region 2 (H2), the heavy chain complementarity determining region 3 (H3), the light chain complementarity determining region 1 (L1), the light chain complementarity determining region 2 (L2), and the light chain complementarity determining region 3 (L3) of the human anti-IL-33 monoclonal antibody is any one of C1 to C5 in Table 1 above,

[0077] The pharmaceutical composition contains substantially no sodium chloride or contains less than 30 mM sodium chloride.

[0078]

[43] Use for the manufacture of a pharmaceutical composition for the treatment or prevention of an IL-33-related disease,

[0079] The pharmaceutical composition contains a human anti-IL-33 monoclonal antibody and substantially no sodium chloride, or contains less than 30 mM sodium chloride.

[0080] Among them, the combination of amino acid sequences of the heavy chain complementarity determining region 1 (H1), the heavy chain complementarity determining region 2 (H2), the heavy chain complementarity determining region 3 (H3), the light chain complementarity determining region 1 (L1), the light chain complementarity determining region 2 (L2), and the light chain complementarity determining region 3 (L3) of the human anti-IL-33 monoclonal antibody is any one of C1 to C5 in Table 1 above.

[0081]

[44] The pharmaceutical composition according to [12-1], wherein the disaccharide is a saccharide selected from the group consisting of sucrose and trehalose, and the sugar alcohol is a saccharide selected from the group consisting of sorbitol and mannitol.

[0082]

[45] The pharmaceutical composition according to [12-1] or

[44] , wherein the disaccharide is sucrose and the sugar alcohol is sorbitol.

[0083]

[46] The pharmaceutical composition according to any one of [1] to

[12] , comprising at least one polyol having a solubility in water at 20°C of 100 g / 100 g or more.

[0084]

[47] The pharmaceutical composition according to

[46] , wherein the polyol having a solubility in water of 100 g / 100 g or more at 20°C is a sugar having a solubility in water of 100 g / 100 g or more at 20°C.

[0085]

[48] ​​The pharmaceutical composition according to

[47] , wherein the sugar having a solubility in water at 20°C of 100 g / 100 g or more is a sugar selected from sorbitol and sucrose.

[0086]

[49] The pharmaceutical composition according to

[15] , wherein the nonionic surfactant is polysorbate 20, polysorbate 80 or poloxamer 188.

[0087]

[50] A pharmaceutical composition comprising a human anti-IL-33 monoclonal antibody as an active ingredient, wherein:

[0088] The combination of amino acid sequences of the heavy chain complementarity determining region 1 (H1), the heavy chain complementarity determining region 2 (H2), the heavy chain complementarity determining region 3 (H3), the light chain complementarity determining region 1 (L1), the light chain complementarity determining region 2 (L2), and the light chain complementarity determining region 3 (L3) of the human anti-IL-33 monoclonal antibody is any one of C1 to C5 in Table 1,

[0089] The pharmaceutical composition contains a buffer, a nonionic surfactant, and a polyol.

[0090]

[51] The pharmaceutical composition according to

[50] , wherein the polyol is a polyol having a solubility in water at 20°C of 100 g / 100 g or more.

[0091]

[52] The pharmaceutical composition of

[50] or

[51] , which contains substantially no sodium chloride or contains less than 30 mM sodium chloride.

[0092] Effects of the Invention

[0093] The pharmaceutical composition of the present invention (or the pharmaceutical composition used in the present invention) containing the human anti-IL-33 monoclonal antibody (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02) is excellent in safety and efficacy because the white turbidity is suppressed. In addition, the pharmaceutical composition of the present invention does not significantly change the pH in the solution state, can be stably stored for a long time, and is suitable for the storage and use of the preparation. BRIEF DESCRIPTION OF THE DRAWINGS

[0094] [ Figure 1 ] Figure 1 The P7N formulation of the A10-1C04 ​​antibody is shown as being turbid.

[0095] [ Figure 2 ] Figure 2 The results show the effect of suppressing white turbidity by adding sugar. The white turbidity of the A10-1C04 ​​antibody was improved in the formulations (A5S, H6S, P6S, and P7S) to which sorbitol was added. DETAILED DESCRIPTION

[0096] In order to facilitate understanding of the present invention, the terms used in the present invention are explained below.

[0097] [Medical composition]

[0098] In the present specification, a pharmaceutical composition refers to a composition prepared in a form that can be administered to animals such as humans. Therefore, a pharmaceutical composition may also refer to a preparation that has been given a dosage form.

[0099] [essentially]

[0100] In the present specification, “substantially containing no sodium chloride” means that no sodium chloride is added to the pharmaceutical composition of the present invention, and means that no sodium chloride is contained at a level that causes white turbidity in the pharmaceutical composition of the present invention.

[0101] [Surfactant]

[0102] In this specification, surfactant refers to a general term for substances having a part (hydrophilic group) that is easily affinity with water and a part (lipophilic group·hydrophobic group) that is easily affinity with oil in the molecule. Any surfactant commonly used in antibody preparations can be used, and as an example, polysorbate 20, polysorbate 80, and poloxamer 188 can be cited.

[0103] [Antibody]

[0104] The term "antibody" in this specification is used in the broadest sense, including human antibodies, humanized antibodies, antibodies derived from non-human species, monoclonal antibodies, and polyclonal antibodies. In addition, the "antibody" in this specification can be a multispecific antibody (e.g., a bispecific antibody), a drug-fused antibody (ADC), or an antigen-binding fragment such as dAbs, scFv, Fab, F(ab)'2, Fab', etc.

[0105] [Monoclonal Antibodies]

[0106] In this specification, the term "monoclonal antibody" when used in this specification refers to an antibody composed of a group of substantially homogeneous antibodies, that is, for example, each antibody constituting the group is the same except for slight differences such as sugar chains and amino acid modifications. Generally, in contrast to polyclonal antibodies containing different antibodies, each monoclonal antibody binds to a single epitope on an antigen. The modifier "monoclonal" indicates that the antibody characteristic is obtained from a group of substantially homogeneous antibodies, and should not be interpreted as requiring the preparation of antibodies by a specific method. For example, the monoclonal antibodies used according to the present invention can be made using the hybridoma method originally described by Kohler et al., Nature, 256: 495 (1975), or can also be made using a recombinant DNA method (e.g., see U.S. Patent No. 4,816,567). "Monoclonal antibodies" can also be isolated from phage antibody libraries using the methods described in, for example, Clackson et al., Nature, 352: 624-628 (1991) and Marks et al., J. Mol. Biol., 222: 581-597 (1991).

[0107] [5 types of human anti-IL-33 monoclonal antibodies]

[0108] The five human anti-IL-33 monoclonal antibodies used in the present invention refer to the five clones of human anti-IL-33 monoclonal antibodies A10-1C04, A23-1A05, A25-2C02, A25-3H04 and A26-1F02 disclosed in International Publication No. 2015 / 099175, which are human anti-IL-33 monoclonal antibodies composed of heavy chains and light chains having the following amino acid sequences.

[0109] (a) A10-1C04:

[0110] Heavy chain:

[0111] EVQLLESGGGLVQPGGSLRLSCAASGFTFSDYYMNWVRQAPGKGLEWVSSISRYSSYIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDIGGMDVWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (Sequence Listing No. 1); and

[0112] · Light chain:

[0113] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAVYDVHWYQQLPGTAPKLLIYRNNQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCQTYDSSRWVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (Sequence Listing No. 2)

[0114] (b) A23-1A05:

[0115] · Heavy chain:

[0116] EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMHWVRQAPGKGLEWVSSISARSRYHYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARLATRHNAFDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (Sequence Listing No. 3); and

[0117] · Light chain:

[0118] QSVLTQPPSASGTPGQRVTISCSGSSSNIGNNAVSWYQQLPGTAPKLLIYASNMRVIGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDDSQKALVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (Sequence Listing No. 4)

[0119] (c) A25-2C02:

[0120] · Heavy chain:

[0121] EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMHWVRQAPGKGLEWVSSISARSSYIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARLATRNNAFDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (Sequence Listing No. 5); and

[0122] · Light chain:

[0123] QSVLTQPPSASGTPGQRVTISCSGSSSNIGRNAVNWYQQLPGTAPKLLIYASNMRVSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCWAWDDSQKVGVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (Sequence Listing No. 6)

[0124] (d) A25-3H04:

[0125] · Heavy chain:

[0126] EVQLLESGGGLVQPGGSLRLSCAASGFTFSRYYMHWVRQAPGKGLEWVSSISAQSSHIYYADSVEGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARLATRQNAFDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (Sequence Listing No. 7); and

[0127] · Light chain:

[0128] QSVLTQPPSASGTPGQRVTISCSGSSSNIGRNAVNWYQQLPGTAPKLLIYASNMRRSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCSAWDDSQKVVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (Sequence Listing No. 8)

[0129] (e) A26-1F02:

[0130] · Heavy chain:

[0131] EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMHWVRQAPGKGLEWVSSISARSSYLYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARLATRHVAFDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (Sequence Listing No. 9); and

[0132] · Light chain:

[0133] QSVLTQPPSASGTPGQRVTISCSGSSSNIGNNAVNWYQQLPGTAPKLLIYASNMRRPGVPDRFSGSKSGTSASLAISGLRSEDEADYYCEAWDDSQKAVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (Sequence Listing No. 10)

[0134] [Complementary Determining Region (CDR)]

[0135] In this specification, complementarity determining region (CDR) refers to the amino acid residues of the antibody related to antigen binding. CDR is also commonly referred to as "hypervariable region", which is the amino acid sequence specific to the molecular species of the antibody, and can be determined by the method of Kabat et al. (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991). CDR has 3 (L1, L2, L3) in the light chain of the antibody, and 3 (H1, H2, H3) in the heavy chain of the antibody. Each CDR of the 5 clones of A10-1C04, A23-1A05, A25-2C02, A25-3H04, and A26-1F02 is shown below.

[0136] (a) A10-1C04:

[0137] H1: DYYMN (sequence number 11 in the sequence list)

[0138] H2: SISRYSSYIYYADSVKG (sequence number 12 in the sequence listing)

[0139] H3:DIGGMDV (SEQ ID NO. 13 in the sequence listing)

[0140] L1:TGSSSNIGAVYDVH (SEQ ID NO. 14 in the sequence listing)

[0141] L2: RNNQRPS (sequence number 15 in the sequence list)

[0142] L3: QTYDSSRWV (sequence number 16 in the sequence list)

[0143] (b) A23-1A05:

[0144] H1: NYYMH (SEQ ID NO. 17 in the sequence listing)

[0145] H2: SISARSRYHYYADSVKG (SEQ ID NO. 18 in the sequence listing)

[0146] H3: LATRHNAFDI (SEQ ID NO. 19 in the sequence listing)

[0147] L1: SGSSSNIGNNAVS (sequence number 20 in the sequence list)

[0148] L2: ASNMRVI (SEQ ID NO. 21 in the sequence listing)

[0149] L3:GAWDDSQKALV (SEQ ID NO. 22 in the sequence listing)

[0150] (c) A25-2C02:

[0151] H1: NYYMH (SEQ ID NO. 17 in the sequence listing)

[0152] H2: SISARSSYIYYADSVKG (SEQ ID NO. 23 in the sequence listing)

[0153] H3: LATRNNAFDI (SEQ ID NO. 24 in the sequence listing)

[0154] L1: SGSSSNIGRNAVN (SEQ ID NO: 25 in the sequence listing)

[0155] L2: ASNMRVS (SEQ ID NO. 26 in the sequence listing)

[0156] L3: WAWDDSQKVGV (SEQ ID NO: 27 in the sequence listing)

[0157] (d) A25-3H04:

[0158] H1: RYYMH (SEQ ID NO. 28 in the sequence listing)

[0159] H2:SISAQSSHIYYADSVEG (SEQ ID NO. 29 in the sequence listing)

[0160] H3: LATRQNAFDI (sequence number 30 in the sequence listing)

[0161] L1: SGSSSNIGRNAVN (SEQ ID NO: 25 in the sequence listing)

[0162] L2: ASNMRRS (SEQ ID NO. 31 in the sequence listing)

[0163] L3: SAWDDSQKVVV (sequence number 32 in the sequence list)

[0164] (e) A26-1F02:

[0165] H1: NYYMH (SEQ ID NO. 17 in the sequence listing)

[0166] H2: SISARSSYLYYADSVKG (SEQ ID NO. 33 in the sequence listing)

[0167] H3: LATRHVAFDI (sequence number 34 in the sequence listing)

[0168] L1: SGSSSNIGNNAVN (sequence number 35 in the sequence list)

[0169] L2: ASNMRRP (SEQ ID NO. 36 in the sequence listing)

[0170] L3:EAWDDSQKAVV (SEQ ID NO. 37 in the sequence listing)

[0171] [Variable region]

[0172] In this specification, the variable region refers to the part other than the constant region of the monoclonal antibody, which is related to the binding with the antigen and is the part that determines the specificity of the antibody that undergoes various changes according to the antigen. The variable region includes a heavy chain variable region and a light chain variable region, and the heavy chain variable region and light chain variable region of the five clones A10-1C04, A23-1A05, A25-2C02, A25-3H04, and A26-1F02 are shown below.

[0173] (a) A10-1C04:

[0174] Heavy chain variable region:

[0175] EVQLLESGGGLVQPGGSLRLSCAASGFTFSDYYMNWVRQAPGKGLEWVSSISRYSSYIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDIGGMDVWGQGTLVTVSS (SEQ ID NO: 38 in the sequence listing)

[0176] Light chain variable region:

[0177] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAVYDVHWYQQLPGTAPKLLIYRNNQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCQTYDSSRWVFGGGTKLTVLG (SEQ ID NO: 39 of the sequence listing)

[0178] (b) A23-1A05:

[0179] Heavy chain variable region:

[0180] EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMHWVRQAPGKGLEWVSSISARSRYHYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARLATRHNAFDIWGQGTLVTVSS (SEQ ID NO: 40 in the sequence listing)

[0181] Light chain variable region:

[0182] QSVLTQPPSASGTPGQRVTISCSGSSSNIGNNAVSWYQQLPGTAPKLLIYASNMRVIGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDDSQKALVFGGGTKLTVLG (SEQ ID NO: 41 of the sequence listing)

[0183] (c) A25-2C02:

[0184] Heavy chain variable region:

[0185] EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMHWVRQAPGKGLEWVSSISARSSYIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARLATRNNAFDIWGQGTLVTVSS (SEQ ID NO: 42 in the sequence listing)

[0186] Light chain variable region:

[0187] QSVLTQPPSASGTPGQRVTISCSGSSSNIGRNAVNWYQQLPGTAPKLLIYASNMRVSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCWAWDDSQKVGVFGGGTKLTVLG (SEQ ID NO: 43 of the sequence listing)

[0188] (d) A25-3H04:

[0189] Heavy chain variable region:

[0190] EVQLLESGGGLVQPGGSLRLSCAASGFTFSRYYMHWVRQAPGKGLEWVSSISAQSSHIYYADSVEGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARLATRQNAFDIWGQGTLVTVSS (SEQ ID NO: 44 in the sequence listing)

[0191] Light chain variable region:

[0192] QSVLTQPPSASGTPGQRVTISCSGSSSNIGRNAVNWYQQLPGTAPKLLIYASNMRRSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCSAWDDSQKVVVFGGGTKLTVLG (SEQ ID NO: 45 of the sequence listing)

[0193] (e) A26-1F02:

[0194] Heavy chain variable region:

[0195] EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMHWVRQAPGKGLEWVSSISARSSYLYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARLATRHVAFDIWGQGTLVTVSS (SEQ ID NO: 46 in the sequence listing)

[0196] Light chain variable region:

[0197] QSVLTQPPSASGTPGQRVTISCSGSSSNIGNNAVNWYQQLPGTAPKLLIYASNMRRPGVPDRFSGSKSGTSASLAISGLRSEDEADYYCEAWDDSQKAVVFGGGTKLTVLG (SEQ ID NO: 47 of the sequence listing)

[0198] "Stablize"

[0199] In this specification, "stable" means that the pharmaceutical composition containing the antibody actually maintains its properties (e.g., physical properties, chemical properties and / or biological activity) after storage. For example, various analytical techniques for determining the stability of proteins represented by antibodies are available in the art, which are summarized in Peptide and Protein Drug Delivery, 247-301, Vincent Lee Ed., Marcel Dekker, Inc., New York, NY, Pubs. (1991) and Jones, A. Adv. Drug Delivery Rev. 10: 29-90 (1993). The stability of the pharmaceutical composition containing the antibody can be evaluated at a selected temperature and for a selected period of time. A "stable" pharmaceutical composition containing an antibody is one in which no significant change is observed at a refrigerated temperature (2 to 8°C) for at least 1 month, 3 months, 6 months, 12 months, preferably 2 years, and more preferably 3 years, or at room temperature (23 to 27°C) for at least 3 months, preferably 6 months, and more preferably 1 year, or under stress conditions (about 40°C or about 50°C) for at least 1 week, 2 weeks, 1 month, preferably 3 months, and more preferably 6 months. Various stability criteria can be used as indicators, such as visual inspection abnormalities (white turbidity, etc.), pH, viscosity, antibody binding to antigen, antibody inhibitory activity against antigen molecules (e.g., IL-33 inhibitory activity against IL-6 production induction), antibody effector function, and antibody degradation.

[0200] “Effector function”

[0201] The "effector function" of an antibody refers to those biological activities that are produced by the Fc region of the antibody (the Fc region of the native sequence or the Fc region of the amino acid sequence mutant). Examples of the effector function of an antibody include C1q binding, complement-dependent cytotoxicity, Fc receptor binding, antibody-dependent cell-mediated cytotoxicity (ADCC), phagocytosis, downregulation of cell surface receptors (e.g., B cell receptor, BCR), etc.

[0202] [White turbidity]

[0203] In this specification, "white turbidity" refers to a white turbid state by visual inspection of color and / or transparency (turbidity). The occurrence of white turbidity can also be analyzed by using a fluid imaging particle analyzer, a particle counter to measure the microparticles, using dynamic light scattering (DLS) to measure the interaction parameter (hereinafter referred to as "interaction parameter" or "Kd value"), and measuring turbidity (absorbance at 650nm (OD650)). As an analytical value related to "white turbidity", for example: the OD650 of the pharmaceutical composition containing the antibody is 0.009, 0.010, 0.011, 0.012, 0.013, or 0.014 or more, the Kd value is 0, -1, -2, -3, or -4mL / g or less, and / or the particles of 1.5μm or more measured by the particle counter are 500, 750, 1000, 1250 or 1500 / mL or more.

[0204] [Freeze-dried preparation]

[0205] In this specification, "lyophilized preparation" refers to a pharmaceutical composition that is almost free of water and has been dried (e.g., freeze-dried). Antibody freeze-drying technology is generally known in the art, for example, see Rey & May (2004) Freeze-Drying / Lyophilization of Pharmaceutical & Biological Products ISBN0824748689.

[0206] [IL-33]

[0207] IL-33 is a cytokine belonging to the IL-1 family, also known as NF-HEV. IL-33 has the following functions: when released as a cytokine to the extracellular space, it binds to the IL-33 receptor (ST2 and IL-1RAcP) and starts intracellular signal transduction in cells expressing the IL-33 receptor. The signal transduction induced by IL-33 is not limited, and there are NF-κB pathways and MAPKKs pathways, which ultimately trigger the production of various cytokines, chemokines, and inflammatory mediators. As examples of cytokines induced by IL-33, TNF-α, IL-1β, IL-3, IL-4, IL-5, IL-6, IL-13, etc. can be cited, and IL-5, IL-6, and IL-13 can be particularly induced. As examples of chemokines induced by IL-33, CXCL2, CCL2, CCL3, CCL6, CCL17, CCL24, etc. can be cited. Examples of inflammatory mediators induced by IL-33 include PGD2 and LTB4. Cytokines, chemokines, and inflammatory mediators induced by IL-33 participate in immune system cell migration, cytokine production, and degranulation, causing inflammation. In the present invention, as long as the "five human anti-IL-33 monoclonal antibodies" bind and inhibit at least one of the aforementioned functions, they may refer to any of full-length IL-33 or mature IL-33, or derivatives or mutants having homology therewith. In addition, it may be human IL-33 or IL-33 derived from other organisms.

[0208] [Pharmaceutically permitted]

[0209] In the present specification, the term "pharmaceutically acceptable" means that the pharmaceutical composition does not interfere with the effectiveness of the biological activity of the active ingredient (which may be plural) and is non-toxic.

[0210] [Isotonic]

[0211] As used herein, an "isotonic" preparation has substantially the same osmotic pressure as human blood. An isotonic pharmaceutical composition generally has an osmotic pressure ratio of about 0.9 to 1.2 relative to blood. The osmotic pressure can be measured using, for example, a vapor pressure type or freezing point type osmometer.

[0212] [pH drift]

[0213] In the present specification, the term "pH drift" means that the pH value of the pharmaceutical composition of the present invention changes before and after treatment such as storage or concentration.

[0214] Hereinafter, embodiments of the present invention will be described. It should be noted that the following embodiments are examples for describing the present invention, and the present invention is not limited to these embodiments.

[0215] The human anti-IL-33 monoclonal antibody according to the present invention can be produced by a known technique, for example, the method described in Patent Document 1.

[0216] The monoclonal antibody produced as described above can be prepared as a desired composition by dialysis, ultrafiltration, or ammonium sulfate precipitation. Alternatively, the monoclonal antibody can be prepared into a solution containing a desired buffer and then added with sugars, surfactants, etc. to prepare a formulation.

[0217] The inventors of the present application have found that when 5 types of human anti-IL-33 monoclonal antibodies are made into solutions, they partially aggregate and become turbid. The turbidity is caused by the formation of microparticles as aggregates containing antibodies. When it is made into a pharmaceutical composition, there are concerns that the biological activity of the antibody as an active ingredient is reduced, the anti-drug antibody (ADA) is induced by the highly immunogenic aggregates, resulting in deterioration of pharmacokinetics, and inflammation is caused by the aggregates themselves. Therefore, it is necessary to suppress turbidity. Therefore, the present invention relates to a pharmaceutical composition in which turbidity is suppressed. The pharmaceutical composition of the present invention may contain 5 types of human anti-IL-33 monoclonal antibodies as active ingredients, and may also contain salts, buffers, surfactants, sugars, etc.

[0218] [Salt concentration]

[0219] The inventors of the present application have found that the main cause of the turbidity of the five human anti-IL-33 monoclonal antibodies is salt. Therefore, the pharmaceutical composition of the present invention preferably has a low salt concentration, preferably 50mM or less, 40mM or less, 30mM or less, 25mM or less, 20mM or less, 15mM or less, 10mM or less, 5mM or less, 3mM or less, 2mM or less, 1mM or less, and more preferably substantially no salt. In addition, the pharmaceutical composition of the present invention preferably has a low salt concentration, preferably less than 50mM, less than 40mM, less than 30mM, less than 25mM, less than 20mM, less than 15mM, less than 10mM, less than 5mM, less than 3mM, less than 2mM, less than 1mM, and more preferably substantially no salt. It should be noted that in the pharmaceutical composition of the present invention, as examples of low concentrations or substantially no salts, inorganic salts or organic salts can be cited. Examples of the inorganic salt include sodium chloride, potassium chloride, magnesium chloride, calcium chloride, sodium sulfate, potassium sulfate, magnesium sulfate, and calcium sulfate, with sodium chloride being preferred.

[0220] [Buffer and pH]

[0221] The pharmaceutical composition of the present invention uses a buffer to adjust the pH. As a suitable buffer for the pharmaceutical composition, there is no limitation, and examples include gluconate, histidine, citrate, phosphate [e.g., sodium or potassium], succinate [e.g., sodium], acetate, trishydroxymethylaminomethane, glycine, arginine and combinations thereof, which can be used separately according to the pH value to be adjusted. The pharmaceutical composition of the present invention preferably contains acetate, histidine, or phosphate as a buffer, and more preferably contains histidine as a buffer. The concentration of the buffer is not particularly limited as long as it is pharmaceutically permitted, and is preferably 1mM to 150mM, more preferably 5mM to 100mM, and further preferably 10mM to 50mM. In addition, the concentration of the buffer is preferably about 1mM, about 5mM, about 10mM, about 15mM, about 20mM, about 25mM, about 30mM, about 35mM, about 40mM, about 45mM or about 50mM. From the viewpoint of using a salt, particularly an inorganic salt, as a buffer, it is preferred to use a concentration of less than 30 mM, particularly 10 mM or less.

[0222] In the pharmaceutical composition of the present invention, histidine (e.g., 5 mM to 50 mM, such as 10 mM to 50 mM, about 5 mM, about 10 mM, about 15 mM, about 20 mM, about 25 mM, about 30 mM, about 35 mM, about 40 mM, about 45 mM, about 50 mM) is particularly useful as a buffer. In one embodiment, the stable pharmaceutical composition contains 5 mM to 20 mM histidine. The pH of the pharmaceutical composition may be in the range of 4.0 to 8.0. A pH in the range of 4.5 to 7.5, for example, 5.0 to 7.0, 5.2 to 6.8, for example, about 4.2, about 4.3, about 4.4, about 4.5, about 4.6, about 4.7, about 4.8, about 4.9, about 5.0, about 5.1, about 5.2, about 5.3, about 5.4, about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, about 6.0, about 6.1, about 6.2, about 6.3, about 6.4, about 6.5, about 6.6, about 6.7, about 6.8, about 6.9, about 7.0, about 7.1, about 7.2, about 7.3, about 7.4, about 7.5, about 7.6, about 7.7, about 7.8 is common. When the pharmaceutical composition of the present invention is at pH 4 or pH 8, the turbidity increases during long-term storage. In addition, when the pharmaceutical composition of the present invention is at pH 4 or pH 8, the pH drift is significant during long-term storage. Therefore, in one embodiment, the pH of the stable pharmaceutical composition containing an antibody is greater than 4 and less than 8, preferably greater than 5 and less than 7, more preferably greater than 5.5 and less than 6.5, and most preferably 6.0.

[0223] [Surfactant]

[0224] The pharmaceutical composition of the present invention preferably contains a surfactant. Suitable surfactants for pharmaceutical compositions are not limited, including nonionic surfactants, ionic surfactants, zwitterionic surfactants and combinations thereof. Common surfactants used in the present invention are not limited, including sorbitan fatty acid esters (e.g., sorbitan monocaprylate, sorbitan monolaurate, sorbitan monopalmitate), sorbitan trioleate, glycerol fatty acid esters (e.g., glycerol monocaprylate, glycerol monomyristate, glycerol monostearate), polyglycerol fatty acid esters (e.g., decapolyglycerol monostearate, decapolyglycerol distearate, decapolyglycerol monolinoleate), polyoxyethylene sorbitan fatty acid esters (e.g., polyoxyethylene sorbitan mono laurate, polyoxyethylene sorbitan monooleate, polyoxyethylene sorbitan monostearate, polyoxyethylene sorbitan monopalmitate, polyoxyethylene sorbitan trioleate, polyoxyethylene sorbitan tristearate), polyoxyethylene sorbitan fatty acid esters (e.g., polyoxyethylene sorbitan tetrastearate, polyoxyethylene sorbitan tetraoleate), polyoxyethylene glycerol fatty acid esters (e.g., polyoxyethylene glycerol monostearate), polyethylene glycol fatty acid esters (e.g., polyethylene glycol distearate), polyoxyethylene alkyl ethers (e.g., For example, polyoxyethylene lauryl ether), polyoxyethylene polyoxypropylene alkyl ether (for example, polyoxyethylene polyoxypropylene glycol, polyoxyethylene polyoxypropylene propyl ether, polyoxyethylene polyoxypropylene hexadecyl ether), polyoxyethylene alkylphenyl ether (for example, polyoxyethylene nonylphenyl ether), polyoxyethylene hydrogenated castor oil (for example, polyoxyethylene castor oil, polyoxyethylene hydrogenated castor oil), polyoxyethylene beeswax derivatives (for example, polyoxyethylene sorbitan beeswax), polyoxyethylene lanolin derivatives (for example, polyoxyethylene lanolin), polyoxyethylene fatty acid amides (for example, polyoxyethylene Stearic acid amide), C10-C18 alkyl sulfates (e.g., sodium hexadecyl sulfate, sodium lauryl sulfate, sodium oleyl sulfate), polyoxyethylene C10-C18 alkyl ether sulfates obtained by adding an average of 2-4 moles of ethylene oxide units (e.g., sodium polyoxyethylene lauryl sulfate), and C1-C18 alkyl sulfosuccinate acid salts (e.g., sodium lauryl sulfosuccinate), as well as natural surfactants, such as lecithin, glycerophospholipids, sphingomyelin (e.g., sphingomyelin), and sucrose esters of C12-C18 fatty acids.

[0225] The pharmaceutical composition of the present invention may contain one or more of the above-mentioned surfactants. Preferred surfactants are nonionic surfactants (e.g., sorbitan fatty acid esters, sorbitan trioleate, glycerol fatty acid esters, polyglycerol fatty acid esters, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene sorbitol fatty acid esters, polyoxyethylene glycerol fatty acid esters, polyoxyethylene alkyl ethers, polyoxyethylene polyoxypropylene alkyl ethers, polyoxyethylene alkylphenyl ethers, polyoxyethylene hydrogenated castor oil, polyoxyethylene beeswax derivatives, polyoxyethylene lanolin derivatives, polyoxyethylene fatty acid amides), more preferably polyoxyethylene alkyl ethers (e.g., poloxamer 188) or polyoxyethylene sorbitan fatty acid esters, such as polysorbate 20, 40, 60 or 80. The concentration of the surfactant may be any concentration commonly used in the art. For example, it may be used at a concentration of about 0.01% (w / v) to about 0.1% (w / v), for example, about 0.01% (w / v) to about 0.04% (w / v), for example, about 0.01% (w / v), about 0.02% (w / v), about 0.04% (w / v), about 0.06% (w / v), about 0.08% (w / v), or about 0.1% (w / v). Among the surfactants, polysorbate 80 (Tween 80) is particularly useful. In one embodiment, the stable pharmaceutical composition contains about 0.02% (w / v) of polysorbate 80. In one embodiment, the stable pharmaceutical composition contains about 0.02% (w / v) of polysorbate 20.

[0226] [Polyol]

[0227] The pharmaceutical composition of the present invention preferably contains a polyol. By adding a polyol, the osmotic pressure of the pharmaceutical composition can be adjusted and the formation of aggregates can be suppressed.

[0228] The polyol contained in the pharmaceutical composition of the present invention is not particularly limited as long as it is pharmaceutically acceptable, but preferably is a polyol that dissolves 100 g or more in 100 g of water at 20°C (ie, a polyol having a solubility in water at 20°C of 100 g / 100 g or more).

[0229] The polyol is also called a polyol and is any molecule having two or more alcoholic hydroxyl groups. Examples thereof include glycerol (glycerin), propylene glycol, polyethylene glycol (PEG), and sugars, and sugars are preferred.

[0230] Suitable sugars for the pharmaceutical composition of the present invention are not limited, and include monosaccharides, disaccharides, trisaccharides, polysaccharides, sugar alcohols, reducing sugars, non-reducing sugars, and the like, and have the general formula (CH 2O)n compounds and derivatives thereof. Examples of sugars include monosaccharides such as glucose, fructose, and galactose; disaccharides such as sucrose, trehalose, lactose, maltose, lactulose, maltulose, isomaltulose, and melibiose; trisaccharides such as melezitose, raffinose, and maltotriose; polysaccharides such as stachyose and dextran; and sugar alcohols such as sorbitol, mannitol, erythritol, maltitol, lactitol, arabitol, and xylitol. Among monosaccharides, disaccharides, and trisaccharides, reducing sugars include glucose, fructose, lactose, maltose, lactulose, maltulose, isomaltulose, melibiose, melezitose, and maltotriose; and non-reducing sugars include trehalose, sucrose, and raffinose. As the sugar contained in the pharmaceutical composition of the present invention, sorbitol, sucrose, trehalose or mannitol is preferred, and sorbitol or sucrose is most preferred.

[0231] The concentration of the saccharide contained in the pharmaceutical composition of the present invention is not particularly limited as long as it is pharmaceutically permitted, and is preferably 50mM to 300mM, more preferably 165mM to 275mM, and further preferably 200mM to 220mM. In addition, the concentration of the saccharide is preferably about 50mM, about 55mM, about 100mM, about 110mM, about 150mM, about 165mM, about 200mM, about 220mM, about 275mM, about 300mM. In one embodiment, the stable pharmaceutical composition contains 3% to 5% (w / v) (165 to 275mM) of sorbitol. In one embodiment, the stable pharmaceutical composition contains about 3.6% (w / v) or about 4% (w / v) (about 200mM or about 220mM, respectively) of sorbitol.

[0232] Preferably, the osmotic pressure ratio of the pharmaceutical composition of the present invention is 0.5 to 4, more preferably 0.7 to 3, further preferably 1 to 2, and most preferably isotonic (0.9 to 1.2), for example, about 0.9, about 1.0, about 1.1, about 1.2, about 1.3, about 1.4, about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, about 2.0. The osmotic pressure of the pharmaceutical composition can be adjusted by the concentration of components added other than the active ingredient, such as salts or polyols. From the viewpoint of reducing the salt concentration, the osmotic pressure of the pharmaceutical composition is preferably adjusted using a polyol.

[0233] [Human anti-IL-33 monoclonal antibody]

[0234] The pharmaceutical composition of the present invention contains a human anti-IL-33 monoclonal antibody (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02) as an active ingredient. The concentration of the human anti-IL-33 monoclonal antibody contained in the pharmaceutical composition of the present invention is not particularly limited as long as it is pharmaceutically acceptable, but is preferably 1 mg / mL to 200 mg / mL, 5 mg / mL to 175 mg / mL, 10 mg / mL to 150 mg / ml, 20 mg / mL to 150 mg / mL, for example, about 1, about 5, about 10, about 20, about 25, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, about 150, about 160, about 170, about 175 mg / ml. The viscosity of the pharmaceutical composition of the present invention is high when the antibody concentration is 175 mg / mL or more, so it is preferably less than 175 mg / mL, and more preferably less than 150 mg / mL. In one embodiment, the stable pharmaceutical composition contains about 10 mg / mL or about 150 mg / mL of A10-1C04. In one embodiment, the stable pharmaceutical composition contains about 10 mg / mL or about 150 mg / mL of A23-1A05. In one embodiment, the stable pharmaceutical composition contains about 10 mg / mL or about 150 mg / mL of A25-2C02. In one embodiment, the stable pharmaceutical composition contains about 10 mg / mL or about 150 mg / mL of A25-3H04. In one embodiment, the stable pharmaceutical composition contains about 10 mg / mL or about 150 mg / mL of A26-1F02.

[0235] The pharmaceutical composition of the present invention preferably has a suitable viscosity, and can be easily administered to any patient. The lower the viscosity, the more it can be administered without force. Regarding the pharmaceutical composition of the present invention, the inventor of the present application found that the viscosity is about 20cP or more, and it is slightly difficult to eject from the syringe when administering. Therefore, the viscosity of the pharmaceutical composition to which the present invention relates is preferably 50cP or less, more preferably 30cP or less, further preferably 20cP or less, and most preferably 10cP or less. In one embodiment, the pharmaceutical composition of the present invention is about 1, about 2, about 5, about 10, about 15, about 20cP. The viscosity of the present invention can be measured using a rheometer (rotational or capillary).

[0236] [Method of using the pharmaceutical composition]

[0237] The pharmaceutical composition of the present invention is used for the treatment (treatment, prevention, etc.) of patients suffering from IL-33-related diseases such as asthma, allergies (atopic dermatitis, hay fever), and endometriosis. The administration method is not particularly limited, and it can be systemically administered or locally administered. For example, intravenous administration, subcutaneous administration, intramuscular administration, and intraperitoneal administration can be mentioned. When the pharmaceutical composition of the present invention is administered subcutaneously, the administration amount is limited, so it is preferred to contain a high concentration of human anti-IL-33 antibodies, for example, preferably a pharmaceutical composition containing 10 mM histidine, 4% (w / v) sorbitol, 0.02% (w / v) polysorbate 80, 150 mg / ml of active ingredients, and the pH is adjusted to 5.5 to 6.5. The pharmaceutical composition of the present invention may contain a low concentration of human anti-IL-33 antibody when administered intravenously, for example, preferably a pharmaceutical composition containing 10 mM histidine, 3.6% (w / v) sorbitol, 0.02% (w / v) polysorbate 80, 10 mg / ml of an active ingredient, and the pH is adjusted to 5.5 to 6.5.

[0238] Examples of IL-33-related diseases include, but are not limited to, asthma, atopic dermatitis, urticaria, hay fever, anaphylactic shock, eosinophilic sinusitis, eosinophilia, Churg-Strauss syndrome, allergic encephalomyelitis, polymyalgia rheumatica, rheumatic heart disease, multiple sclerosis, arthritis (e.g., rheumatoid arthritis, juvenile arthritis, psoriatic arthritis, osteoarthritis, Reiter's syndrome, etc.), systemic lupus erythematosus (including discoid lupus), psoriasis, ankylosing spondylitis, hepatitis (e.g., autoimmune hepatitis, chronic active hepatitis, etc.), inflammatory bowel disease (e.g., ulcerative colitis, Crohn's disease, gluten-sensitive enteropathy, etc.), systemic lupus erythematosus, Sjögren's syndrome, Behcet's disease, Herpes, pemphigoid, autoimmune hemolytic anemia, autoimmune inflammatory eye disease, autoimmune neonatal thrombocytopenia, autoimmune neutropenia, autoimmune oophoritis and orchitis, autoimmune thrombocytopenia, autoimmune thyroiditis, polymyositis, dermatomyositis, myasthenia gravis, adrenergic agonist resistance, alopecia areata, antiphospholipid syndrome, adrenal autoimmune diseases (e.g., autoimmune Addison's disease, etc.), celiac disease-dermatitis, chronic fatigue Immune dysfunction syndrome (CFIDS), cold agglutinin disease, essential mixed globulinemia, fibromyalgia-fibromyositis, glomerulonephritis (e.g., IgA nephropathy (nephrophathy), Graves' disease, hyperthyroidism (i.e., Hashimoto's thyroiditis), idiopathic thrombocytopenic purpura (ITP), mixed connective tissue disease, type 1 or immune-mediated diabetes mellitus, pernicious anemia, polychrondritis, polyglandular syndrome, generalized myotonia syndrome, vitiligo , sarcoidosis, polyendocrine syndrome, other endocrine insufficiency, arteriosclerosis, liver fibrosis (e.g., primary biliary cirrhosis, etc.), pulmonary fibrosis (e.g., idiopathic pulmonary fibrosis, etc.), chronic obstructive pulmonary disease, scleroderma (including CREST syndrome, Raynaud's phenomenon, etc.), endometriosis, adenomyosis, tubulointerstitial nephritis, dense deposit disease, acute kidney injury, myocarditis, cardiomyopathy, neuritis (e.g., Guillain-Barré syndrome, etc.), polyarteritis nodosa, cardiotomy syndrome, chronic inflammatory demyelinating polyneuropathy, IgA neuropathy, lichen planus, Meniere's syndrome, post-myocardial infarction (post-MI), uveitis, uveitis opthalmia, vasculitis, primary agammaglobulinemia, cancer (e.g.,Brain tumor, laryngeal cancer, oral cancer, hypopharyngeal cancer, thyroid cancer, esophageal cancer, breast cancer, lung cancer, stomach cancer, adrenocortical cancer, bile duct cancer, gallbladder cancer, liver cancer, pancreatic cancer, bladder cancer, colon cancer, uterine cancer, ovarian cancer, prostate cancer, testicular cancer, chronic lymphocytic leukemia, chronic myeloid leukemia, Ewing sarcoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, melanoma, mesothelioma, multiple myeloma, etc.), infection resistant to rejection from the immune system (for example, severe acute respiratory syndrome (SARS)), lethal cytokine storm associated with highly toxic influenza infection, and scurvy, preferably asthma, atopic dermatitis, hay fever, anaphylactic shock, scleroderma, Crohn's disease, ulcerative colitis, arthritis, systemic lupus erythematosus, ankylosing spondylitis, liver fibrosis, pulmonary fibrosis, acute kidney injury, vasculitis and cancer, etc.,

[0239] The "stable" pharmaceutical composition of the present invention does not show significant changes at refrigerated temperature (2-8°C) for at least 12 months, preferably 2 years, more preferably 3 years, or at room temperature (22-28°C) for at least 3 months, preferably 6 months, more preferably 1 year. For example, after 2 years of storage at 5°C, no turbidity is observed, OD650 is 0.014 or less, preferably 0.01 or less, more preferably 0.008 or less, pH drift is 1 or less, preferably 0.8 or less, more preferably 0.5 or less, Kd value is -4 mL / g or more, preferably -2 mL / g or more, more preferably a positive value, or the number of particles of 1.5 μm or more measured by a particle counter is 1500 or less, preferably 1000 or less, more preferably 750 or less, and most preferably 500 or less.

[0240] The pharmaceutical composition of the present invention may further contain a preservative, an anti-adsorption agent, an analgesic, a sulfur-containing reducing agent, an antioxidant, and the like as needed.

[0241] The preservative is not particularly limited as long as it is pharmaceutically acceptable, and examples thereof include methyl parahydroxybenzoate, ethyl parahydroxybenzoate, sorbic acid, phenol, cresol, and chlorocresol.

[0242] The anti-adsorption agent is not particularly limited as long as it is pharmaceutically acceptable, and examples thereof include human serum albumin, lecithin, dextran, ethylene oxide-propylene oxide copolymer, hydroxypropyl cellulose, methyl cellulose, polyoxyethylene hydrogenated castor oil, and polyethylene glycol.

[0243] The analgesic is not particularly limited as long as it is pharmaceutically acceptable, and examples thereof include local anesthetics such as lidocaine.

[0244] The sulfur-containing reducing agent is not particularly limited as long as it is pharmaceutically acceptable, and examples thereof include substances having a thiol group such as N-acetylcysteine, N-acetylhomocysteine, lipoic acid, thiodiethylene glycol, thioethanolamine, thioglycerol, thiosorbitol, thioglycolic acid and its salts, sodium thiosulfate, glutathione, and thioalkanoic acids having 1 to 7 carbon atoms.

[0245] The antioxidant is not particularly limited as long as it is pharmaceutically acceptable, and examples thereof include isoascorbic acid, butylated hydroxytoluene, butylated hydroxyanisole, α-tocopherol, tocopherol acetate, L-ascorbic acid and its salts, L-ascorbyl palmitate, L-ascorbyl stearate, sodium bisulfite, sodium sulfite, tripentyl gallate, propyl gallate, or chelating agents such as disodium ethylenediaminetetraacetate (EDTA), sodium pyrophosphate, and sodium metaphosphate.

[0246] The pharmaceutical composition of the present invention may be a freeze-dried preparation for longer-term storage.

[0247] The present invention will be described in more detail by the following examples, but the scope of the present invention is not limited to these examples.

[0248] [Example]

[0249] Comparative Example 1: White turbidity of a pharmaceutical composition containing human anti-IL-33 monoclonal antibody

[0250] Human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04, or A26-1F02) were prepared in a solvent (10 mM sodium phosphate / pH 7 / 150 mM NaCl / 0.02% (w / v) polysorbate 80 (Polysobate 80)) (hereinafter referred to as "P7N") to give a concentration of 150 mg / mL, and the properties were confirmed by naked eyes, and white turbidity was observed (for example, for A10-1C04, shown in Figure 1 ). It should be noted that the presence or absence of white turbidity was confirmed by naked eye observation under white light (13W fluorescent lamp) on a black background. In addition, for the pharmaceutical composition, FlowCam (manufactured by Fluid Imaging Technologies) was used to measure insoluble particles (microparticles of 5 μm or more), and 1919 microparticles / mL were detected. Based on the above results, it is believed that the human anti-IL-33 monoclonal antibody (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02) has high cohesion and is difficult to formulate.

[0251] Comparative Example 2: Effect of pH on the turbidity of a pharmaceutical composition containing a human anti-IL-33 monoclonal antibody

[0252] In order to eliminate the turbidity of human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02), the pH of the pharmaceutical composition was changed by changing the buffer as follows based on the formulation of Comparative Example 1. The antibody concentration was set to 150 mg / ml. The formulation after the study is as follows.

[0253] 10 mM sodium acetate / pH 4 / 150 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "A4N")

[0254] 10 mM sodium acetate / pH 5 / 150 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "A5N")

[0255] 10 mM histidine / pH 6 / 150 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "H6N")

[0256] 10 mM sodium phosphate / pH 6 / 150 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "P6N")

[0257] 10 mM sodium phosphate / pH 7 / 150 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "P7N") and

[0258] 10 mM sodium phosphate / pH 8 / 150 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "P8N")

[0259] The properties were evaluated and as a result, all the formulations with pH 4, 5, 6, 7, and 8 were turbid. Based on this result, it is thought that the turbidity will not improve even if the pH of the formulation is changed.

[0260] In addition, the properties were confirmed by naked eyes by the method described in Comparative Example 1.

[0261] Example 1: Effect of suppressing turbidity by adding sugar

[0262] In order to eliminate the white turbidity of human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02), the following formulations were evaluated by adding 5% sorbitol instead of sodium chloride contained in A4N, A5N, H6N, P6N, P7N and P8N as the formulation of Comparative Example 2. The antibody concentration was set to 150 mg / ml.

[0263] 10 mM sodium acetate / pH 4 / 5% (w / v) sorbitol / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "A4S")

[0264] 10 mM sodium acetate / pH 5 / 5% (w / v) sorbitol / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "A5S")

[0265] 10 mM histidine / pH 6 / 5% (w / v) sorbitol / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "H6S")

[0266] 10 mM sodium phosphate / pH 6 / 5% (w / v) sorbitol / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "P6S")

[0267] 10 mM sodium phosphate / pH 7 / 5% (w / v) sorbitol / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "P7S")

[0268] 10 mM sodium phosphate / pH 8 / 5% (w / v) sorbitol / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "P8S")

[0269] The properties and OD650 (turbidity) were evaluated. As a result, the turbidity was improved by adding sorbitol instead of sodium chloride (for example, for A10-1C04, shown in Figure 2 ). In addition, the addition of sorbitol suppressed the increase in OD650 even after 3 months at 40°C, and the inhibitory effect was particularly good at pH 5 to 7 (Table 3).

[0270] The properties confirmed by naked eyes were observed by the method described in Comparative Example 1. For OD650, 100 μL of the solution of each formulation was measured for absorbance at 650 nm using a Molecular Device microplate reader (SoftMaxPro software).

[0271] [Table 3]

[0272] Table 3: OD650 of various formulations

[0273] formula During preparation After 3 months storage at 40℃ A4S 0.006 0.014 A5S 0.006 0.005 H6S 0.007 0.008 P6S 0.005 0.004 P7S 0.006 0.004 P8S 0.007 0.014

[0274] Example 2: Salt concentration and turbidity (1)

[0275] The interaction parameter (Kd value) as an index of cohesion was measured when human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02) were dissolved in 10 mM histidine / pH 6.0 or pH 5.5 buffer and 0, 50, 100 mM NaCl was added. The measurement temperature was set to 25°C, and the antibody concentration was prepared in a manner of 0.5, 1, 2.5, 5, 10, 20 mg / mL. As a result, when the amount of NaCl added was 50 mM or more, the Kd value was negative (cohesion increased), so it is considered that the amount of NaCl added should be less than 50 mM (for example, for A10-1C04, shown in Table 4). It should be noted that the Kd value can be calculated as follows: the diffusion coefficient (Dm) is obtained by dynamic light scattering, the slope is obtained from a graph of antibody concentration (horizontal axis) and diffusion coefficient (vertical axis), and the slope is divided by the diffusion coefficient (D0) when the concentration is 0. That is, the Kd value is calculated by the following relationship. When the Kd value is a positive number and the larger it is, the lower the cohesion.

[0276] Dm = D0 (1 + Kd value × [antibody concentration])

[0277] [Table 4]

[0278] Table 4: Salt concentration and interaction parameters (unit: mL / g)

[0279] Salt concentration pH5.5 pH6 0mM 19.2 0.17 50mM -4.7 -6.2 100mM -6.4 -7.6

[0280] Example 3: Salt concentration and turbidity (2)

[0281] 0, 5, 10, 30 mM NaCl was added to 10 mM histidine / pH 6.0 / 3.6% (w / v) sorbitol, and the Kd value as an index of cohesion was measured. 2.5, 5, 10, 14 mg / mL of A10-1C04 ​​was used as the antibody. As a result, the Kd value was negative when the amount of NaCl added was 30 mM or more (the Kd values ​​of 0, 5, 10 and 30 mM NaCl were 17.3, 7.3, 1.2 and -4.0 mL / g, respectively), so it is believed that the amount of NaCl added should be less than 30 mM. The Kd value was calculated by the method described in Example 2.

[0282] Example 4: pH Drift

[0283] The pH shift of pharmaceutical compositions containing human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02) during long-term storage was measured. The antibody concentration was set to 150 mg / mL, and formulations with pH 4 to 8 were investigated (A4S, A5S, H6S, P6S, P7S, P8S). For each preparation, the pH was measured during preparation and after being stored at 40°C for 2, 4, 8, and 12 weeks. As a result, the pH drift was significant in formulations of pH 4 (A4S) and pH 8 (P8S) (for example, for A10-1C04, shown in Table 5). Therefore, it is believed that formulations with less pH drift of pH 5 to 7 are better.

[0284] [Table 5]

[0285] Table 5: pH drift when stored at 40°C

[0286] formula During preparation 2 weeks 4 weeks 8 weeks 12 weeks pH Drift A4S 4.0 4.6 5.0 5.0 5.1 +1.1 A5S 5.0 5.3 5.5 5.5 5.6 +0.6 H6S 6.0 6.2 6.3 6.3 6.4 +0.4 P6S 6.1 6.2 6.2 6.3 6.3 +0.2 P7S 7.0 6.8 6.8 6.7 6.8 -0.2 P8S 8.1 7.6 7.2 7.2 7.3 -0.8

[0287] Example 5: Antibody concentration and viscosity

[0288] Pharmaceutical compositions were prepared in such a manner that the antibody concentration of human anti-IL-33 monoclonal antibody (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02) was 15, 50, 100, 125, 150, 175, 200 mg / ml in a formulation of 10 mM histidine / pH 6 / 3.6% (w / v) sorbitol / 0.02% (w / v) polysorbate 80. The viscosity at a measurement temperature of 25° C. was measured using a viscometer (Brookfield DV3TLVCJ0 viscometer (CPA-40Z rotor, CPA-44YZ sample cup)). The viscosity of A10-1C04 ​​at antibody concentrations of 15, 50, 100, 125, 150, 175, and 200 mg / mL was 1.19, 1.84, 5.33, 9.15, 16.29, 47.98, and 84.96 cP, respectively, and it was found that the viscosity increased sharply after exceeding 150 mg / mL. In addition, solutions of various viscosities were prepared and the ejection properties of the syringe were investigated. It was found that administration became slightly difficult after the viscosity exceeded about 20 cP, and it was considered that the antibody concentration should be less than 175 mg / ml.

[0289] Example 6: Addition of sugar and evaluation of cohesion and microparticle generation inhibition effect

[0290] Compositions of 10 mM histidine / pH 6.0 of human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04, or A26-1F02) were prepared at antibody concentrations of 0.6125, 1.25, 2.5, 5, and 10 mg / mL, and the effect of adding sucrose or sorbitol on the Kd value, which is an index of cohesion, was confirmed. The Kd value was calculated by the method described in Example 2. For example, the Kd value when A10-1C04 ​​was used was 33.4 mL / g when no sugar was added, and 24.7 and 33.5 mL / g, respectively, when 3.6% (w / v) sucrose or sorbitol was added, which were all positive values. Based on this result, it is considered that sucrose can also be added instead of sorbitol. In addition, for the formulation (10 mM histidine / pH 6.0 / 3.6% (w / v) sorbitol) with an antibody concentration of 10 mg / ml and the addition of 3.6% (w / v) sorbitol, a particle counter HIAC (System 9703+ manufactured by HACH) was used to measure the particles of 1.5 μm or more during preparation and when stored at 50°C for 1 week. For the measurement using HIAC, each specimen was measured 4 times with an injection volume of 100 μL, and the data of the first time were discarded. As a result, compared with the case where 3.6% (w / v) sorbitol was added, the increase in the number of particles was suppressed, and it is believed that the addition of sorbitol has an aggregation inhibitory effect (for example, for A10-1C04, shown in Table 6).

[0291] [Table 6]

[0292] Table 6: Sorbitol particle generation inhibition effect (unit: particles / 100 μL)

[0293]

[0294] Example 7: Aggregation inhibition effect of surfactant

[0295] A 10 mg / mL composition of human anti-IL-33 monoclonal antibody (A10-1C04, A23-1A05, A25-2C02, A25-3H04, or A26-1F02) in 10 mM histidine / pH 6 / 3.6% (w / v) sorbitol was prepared, and the effect of adding 0.02% (w / v) polysorbate 20 or polysorbate 80 as a surfactant on cohesion was confirmed. Regardless of whether a surfactant was added, no cohesion was observed during the preparation.

[0296] Example 8: Stability of Subcutaneous and Intravenous Formulations

[0297] An intravenous preparation (10 mg / mL antibody / 10 mM histidine / pH 6 / 3.6% (w / v) sorbitol / 0.02% (w / v) polysorbate 80) and a subcutaneous preparation (150 mg / mL antibody / 10 mM histidine / pH 6 / 4% (w / v) sorbitol / 0.02% (w / v) polysorbate 80) of a human anti-IL-33 monoclonal antibody (A10-1C04, A23-1A05, A25-2C02, A25-3H04, or A26-1F02) were prepared. The intravenous preparation was investigated and it was confirmed that even after storage at 5°C for 24 months, no turbidity, increase in the number of aggregated particles, or pH drift was observed, indicating that the preparation was stable. The stability of the subcutaneous preparation was also evaluated in the same manner.

[0298] Example 9: Effect of pH on the turbidity of a pharmaceutical composition containing human anti-IL-33 monoclonal antibody (2)

[0299] In order to eliminate the white turbidity of human anti-IL-33 monoclonal antibody (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02), the following formulation was prepared within the pH range of the formulation confirmed in Comparative Example 2 and without containing sodium chloride.

[0300] 10 mM sodium acetate / pH 4 / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "A4")

[0301] 10 mM sodium acetate / pH 5 / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "A5")

[0302] 10 mM histidine / pH 6 / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "H6")

[0303] 10 mM histidine / pH 7 / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "H7")

[0304] 10 mM sodium phosphate / pH 8 / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "P8")

[0305] For example, the properties of 150 mg / mL A10-1C04 ​​were evaluated, and the results showed that the turbidity was improved by not adding sodium chloride. In addition, the inhibitory effect was particularly good at pH 4 to 7 (Table 7).

[0306] [Table 7]

[0307] Table 7: Properties of various formulations

[0308] formula Characteristics A4 - A5 - H6 - H7 - P8 +

[0309] ---------------------

[0310] -: Clear (no turbidity)

[0311] +: White turbidity occurs

[0312] Example 10: Effect of salt on white turbidity of a pharmaceutical composition containing human anti-IL-33 monoclonal antibody (2)

[0313] In order to eliminate the white turbidity of human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02), various concentrations of sodium chloride were added to the formulations A4, A5, and H6 of Example 9 and evaluated.

[0314] 10 mM sodium acetate / pH 4 / 10 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "A4N10")

[0315] 10 mM sodium acetate / pH 4 / 30 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "A4N30")

[0316] 10 mM sodium acetate / pH 4 / 50 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "A4N50")

[0317] 10 mM sodium acetate / pH 4 / 100 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "A4N100")

[0318] 10 mM sodium acetate / pH 5 / 10 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "A5N10")

[0319] 10 mM sodium acetate / pH 5 / 30 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "A5N30")

[0320] 10 mM sodium acetate / pH 5 / 50 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "A5N50")

[0321] 10 mM sodium acetate / pH 5 / 100 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "A5N100")

[0322] 10 mM histidine / pH 6 / 10 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "H6N10")

[0323] 10 mM histidine / pH 6 / 30 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "H6N30")

[0324] 10 mM histidine / pH 6 / 50 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "H6N50")

[0325] 10 mM histidine / pH 6 / 100 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "H6N100")

[0326] For example, when properties of 150 mg / mL A10-1C04 ​​were evaluated, it was found that under the conditions of pH 4 to 6, white turbidity was suppressed at 10 mM sodium chloride, but white turbidity was observed at more than 30 mM sodium chloride (Table 8).

[0327] [Table 8]

[0328] Table 8: Properties of various formulations

[0329] none N10 N30 N50 N100 A4 - - + + + A5 - - + + + H6 - - + + +

[0330] -: Clear (no turbidity)

[0331] +: White turbidity occurs

[0332] Example 11: Aggregation inhibition effect of surfactant (2)

[0333] In order to confirm the effect of surfactant on 10 mg / mL of human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02), the following formulations with added surfactants were prepared. Then, these specimens were rotated at 30 rpm in a constant temperature bath at 50 ° C using a rotator. The specimens taken out over time were visually inspected for 5 seconds each under a fluorescent lamp of 2000-3750 Lux with a white board and a black board as the background. In the system without the addition of surfactant, a large amount of coagulants was observed after 2 days, while the specimens with the addition of polysorbate 80 and poloxamer 188 still had no coagulants observed after 7 days, especially in the system with the addition of 0.02% or more of polysorbate 80, no coagulants were observed after 14 days. In the specimen to which polysorbate 20 was added, no foreign matter was observed even after 4 days, and although a small amount of foreign matter was observed after 7 days, no foreign matter was observed after 14 days. For example, the results of the test for A10-1C04 ​​are shown in Table 9.

[0334] 10 mM histidine / pH 6 (hereinafter referred to as "H6(-)")

[0335] 10 mM histidine / pH 6 / 0.02% (w / v) poloxamer 188 (hereinafter referred to as "H6PX")

[0336] 10 mM histidine / pH 6 / 0.02% (w / v) polysorbate 20 (hereinafter referred to as "H6P20")

[0337] 10 mM histidine / pH 6 / 0.01% (w / v) polysorbate 80 (hereinafter referred to as "H6PS1")

[0338] 10 mM histidine / pH 6 / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "H6PS2")

[0339] 10 mM histidine / pH 6 / 0.05% (w / v) polysorbate 80 (hereinafter referred to as "H6PS5")

[0340] [Table 9]

[0341] Table 9: Agglomerates observed in samples stored at 50°C and 30 rpm

[0342] Day 0 1 day 2 days 3 days 7 days 14 days H6(-) - - ++ ++ ++ ++ H6PX - - - - - + H6P20 - - - -* + - H6PS1 - - - - - + H6PS2 - - - - - - H6PS5 - - - - - -

[0343] -: No condensation

[0344] +: A small amount of condensation was confirmed

[0345] ++: Confirmed that a large amount of condensation

[0346] *: Remove after 4 days

[0347] Example 12: Addition of sugar and evaluation of cohesion (2)

[0348] A 10 mg / mL composition of 10 mM histidine / pH 6.0 of human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02) was prepared, and mannitol, trehalose, sucrose or sorbitol were added to prepare a drug solution. Then, these specimens were stored at -80°C for more than 8 hours to freeze them, and then placed at room temperature for more than 4 hours to thaw them. Repeat the above operation 6 times. Then, visually inspect for 5 seconds each with a white board and a black board as the background under a fluorescent light of 2000-3750 Lux. By adding sugar, the amount of coagulation after freeze-thaw is reduced compared to the system without addition, and the inhibitory effect of coagulation is particularly shown in the system using sorbitol and sucrose. For example, the results of the implementation of A10-1C04 ​​are shown in Table 10.

[0349] 10 mM histidine / pH 6 (hereinafter referred to as "H6(-)")

[0350] 10 mM histidine / pH 6 / 3.0% (w / v) sorbitol (hereinafter referred to as "H6So3")

[0351] 10 mM histidine / pH 6 / 3.6% (w / v) sorbitol (hereinafter referred to as "H6So3.6")

[0352] 10 mM histidine / pH 6 / 4.0% (w / v) sorbitol (hereinafter referred to as "H6So4")

[0353] 10 mM histidine / pH 6 / 5.0% (w / v) sorbitol (hereinafter referred to as "H6So5")

[0354] 10 mM histidine / pH 6 / 3.6% (w / v) sucrose (hereinafter referred to as "H6Su")

[0355] 10 mM histidine / pH 6 / 3.6% (w / v) trehalose (hereinafter referred to as "H6Tr")

[0356] 10 mM histidine / pH 6 / 3.6% (w / v) mannitol (hereinafter referred to as "H6Ma")

[0357] [Table 10]

[0358] Table 10: Agglomerates observed by visual inspection after freeze-thaw

[0359]

[0360] -: No condensation

[0361] +: A small amount of condensation was confirmed

[0362] ++: Confirmed that a large amount of condensation

[0363] Example 13: Freeze Drying

[0364] A composition of 150 mg / mL of human anti-IL-33 monoclonal antibody (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02) in 10 mM histidine / pH 6.0 / 0.02% (w / v) polysorbate 80 was prepared, and after adding mannitol, trehalose, sucrose or sorbitol, freeze-dried by shelf freeze drying (manufactured by Kyowa Vacuum). For properties, the shape of the cake after freeze drying was visually confirmed.

[0365] For re-solubility, water for injection was added and the solution was left to stand at 5°C for half a day to confirm whether there were any lumps remaining in the solution. The lumps of any sample were good, and re-dissolution after adding water for injection was confirmed. It was confirmed that it could be prepared in the form of a freeze-dried preparation. For example, the results of A10-1C04 ​​are shown in Table 11.

[0366] 10 mM histidine / pH 6 / 0.02% (w / v) polysorbate 80 / 4.0% (w / v) sorbitol (hereinafter referred to as "LYSO")

[0367] 10 mM histidine / pH 6 / 0.02% (w / v) polysorbate 80 / 4.0% (w / v) sucrose (hereinafter referred to as "LYSU")

[0368] 10 mM histidine / pH 6 / 0.02% (w / v) polysorbate 80 / 4.0% (w / v) trehalose (hereinafter referred to as "LYTR")

[0369] 10 mM histidine / pH 6 / 0.02% (w / v) polysorbate 80 / 2.0% (w / v) mannitol (hereinafter referred to as "LYMA")

[0370] [Table 11]

[0371] Table 11: Physical properties after freeze drying

[0372] LYSO LYSU LYTR LYMA Appearance good good good good Resolubility Sol Sol Sol Sol

[0373] Good: clumps formed, Poor: no clumps formed

[0374] Sol: dissolve, Dis: not dissolve

[0375] Example 14: Stability test (1)

[0376] A composition of 10 mM histidine / pH6 / 0.02% (w / v) polysorbate 80 / 3.6% (w / v) sorbitol of 10 mg / mL human anti-IL-33 monoclonal antibody (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02) was prepared, filled into glass vials, sealed with halogenated butyl rubber stoppers, and stored at 2-8°C for 1, 2 and 3 years as a long-term stability test. Visually inspect for 5 seconds each with a white board and a black board as the background under a fluorescent light of 2000-3750 Lux to confirm the presence or absence of aggregation. Binding activity was evaluated by the following method. Human IL-33 was added to a 96-well plate and solid-phased overnight. After blocking with BSA, a sample solution was added to each well to react the HRP-labeled anti-human IgG antibody, and color was developed using TMB. Then, a microplate detector (Molecular Device) was used to measure the absorbance at 450nm and 650nm to obtain EC50. The EC50 of the standard solution measured in the same manner was obtained and its ratio was calculated. Aggregation and pH drift were not observed at any measurement point, and the binding activity of the antibody was not reduced. For example, the results of A10-1C04 ​​are shown in Table 12.

[0377] [Table 12]

[0378] Table 12: Long-term stability test (1)

[0379] 2～8℃T0 2~8℃1 year 2~8℃2 years 2~8℃ 3 years Visual inspection - - - - Binding activity 100％ 110％ 101％ 101％ pH 6.0 6.0 6.0 6.0

[0380] -: No condensation

[0381] +: A small amount of condensation was confirmed

[0382] ++: Confirmed that a large amount of condensation

[0383] Example 15: Stability Test (2)

[0384] A composition of 10 mM histidine / pH 6 / 0.02% (w / v) polysorbate 80 / 4.0% (w / v) sorbitol of 150 mg / mL human anti-IL-33 monoclonal antibody (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02) was prepared, filled into a glass vial, sealed with a halogenated butyl rubber stopper, and stored at 2 to 8°C for 3 months and 6 months as a long-term stability test. Evaluation was performed by the method of Example 14. No aggregation or pH drift was observed at any measurement point, and the binding activity of the antibody was not reduced. For example, the results of the implementation for A10-1C04 ​​are shown in Table 13.

[0385] [Table 13]

[0386] Table 13: Long-term stability test (2)

[0387] 2～8℃T0 2~8℃ 3 months 2~8℃ 6 months Visual inspection - - - Binding activity 115％ 110％ 100％ pH 5.9 5.8 5.8

[0388] -: No condensation

[0389] +: A small amount of condensation was confirmed

[0390] ++: Confirmed that a large amount of condensation

Claims

1. A pharmaceutical composition comprising a human anti-IL-33 monoclonal antibody as an active ingredient, wherein: The human anti-IL-33 monoclonal antibody is (a) A10-1C04 ​​consisting of a heavy chain having an amino acid sequence of SEQ ID NO: 1 and a light chain having an amino acid sequence of SEQ ID NO: 2, The pH of the pharmaceutical composition is adjusted to 5 or more and 7 or less, the concentration of the active ingredient is 10 mg / ml or more and less than 175 mg / ml, the pharmaceutical composition contains 3 to 5 w / v% of sorbitol or sucrose as a polyol, wherein the pH is adjusted using a buffer such as acetate, histidine or phosphate, The pharmaceutical composition contains 0.01 to 0.1 w / v% of polysorbate 20, polysorbate 80 or poloxamer 188 as a surfactant. The pharmaceutical composition does not contain sodium chloride.

2. The pharmaceutical composition according to claim 1, comprising 0.01 to 0.05 w / v% of polysorbate 80 or poloxamer 188 as a surfactant.

3. The pharmaceutical composition according to claim 1, comprising 3 to 5 w / v% of sorbitol as a polyol, wherein The pH is adjusted with histidine, and the pharmaceutical composition contains 0.02 to 0.05 w / v% of polysorbate 80 as a surfactant. 4 . The pharmaceutical composition according to claim 1 , comprising 5 mM to 20 mM of a buffer. The pharmaceutical composition according to claim 1 , which is for subcutaneous administration. The pharmaceutical composition according to claim 1 , which is for intravenous administration.

7. A freeze-dried preparation of the pharmaceutical composition according to any one of claims 1 to 6.

8. The pharmaceutical composition according to claim 1, comprising a human anti-IL-33 monoclonal antibody as an active ingredient, wherein The human anti-IL-33 monoclonal antibody is A10-1C04, The pharmaceutical composition contains 5 mM to 20 mM histidine, 3 to 5 w / v% sorbitol, 0.01 to 0.04 w / v% polysorbate 80, and 10 mg / mL to 150 mg / ml of an active ingredient, does not contain sodium chloride, and has a pH adjusted to 5.5 to 6.

5.

9. The pharmaceutical composition according to claim 8, wherein The human anti-IL-33 monoclonal antibody is A10-1C04, The pharmaceutical composition contains 10 mM histidine, 4 w / v% sorbitol, 0.02 w / v% polysorbate 80, and 150 mg / ml of an active ingredient, does not contain sodium chloride, and has a pH adjusted to 5.5 to 6.

5.

10. The pharmaceutical composition according to claim 8, wherein The human anti-IL-33 monoclonal antibody is A10-1C04, The pharmaceutical composition contains 10 mM histidine, 3.6 w / v% sorbitol, 0.02 w / v% polysorbate 80, and 10 mg / ml of an active ingredient, does not contain sodium chloride, and has a pH adjusted to 5.5 to 6.5.

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