Pharmaceutical composition containing human anti-IL-33 monoclonal antibody

Abstract

The present invention relates to a pharmaceutical composition containing a human anti-IL-33 monoclonal antibody. The purpose of the present invention is to provide a pharmaceutical composition containing a human anti-IL-33 monoclonal antibody suitable for administration to a subject. The present invention provides a pharmaceutical composition containing a human anti-IL-33 monoclonal antibody as an active ingredient, said pharmaceutical composition containing substantially no sodium chloride or less than 30 mM of sodium chloride. In addition, the present invention provides a freeze-drying agent for the pharmaceutical composition.

Description

[0001] This application is a divisional application of a Chinese patent application with an application date of September 13, 2019, an application number of No. 201980060117.X (international application number PCT / JP2019 / 036239), and an invention title of "Pharmaceutical Composition Containing Human Anti-IL-33 Monoclonal Antibody". Technical Field

[0002] 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 particularly to a pharmaceutical composition containing an antibody with improved storage stability such as suppression of turbidity and improvement of pH. Background Art

[0003] In recent years, various pharmaceutical compositions containing antibodies have been developed for practical use, and many pharmaceutical compositions containing antibodies are used as intravenous injection pharmaceutical compositions. On the other hand, due to the needs of the medical field, the demand for developing a pharmaceutical composition containing an antibody as a self-injectable subcutaneous injection pharmaceutical composition has been gradually increasing.

[0004] When designing a pharmaceutical composition containing an antibody for subcutaneous injection, the amount of antibody administered each time is relatively large (about 100 to 200 mg). On the other hand, there is generally a limit on the injection volume in subcutaneous injection, so it is necessary to make the antibody in the administered solution at a high concentration. Therefore, a pharmaceutical composition containing a high concentration of antibody using a freeze-drying concentration technique (i.e., preparing a pharmaceutical composition containing a high concentration of antibody by redissolving a lyophilized pharmaceutical composition with less water than the volume before freeze-drying) is mostly used.

[0005] IgG antibody, which is one of the antibody subtypes commonly used in pharmaceutical applications, is a high-molecular-weight glycoprotein of about 150 kDa. Mainly because the structures (amino acid sequences, sugar chain structures) of the variable regions of the antibody are very diverse, different physical properties are shown depending on the molecular species of the antibody.

[0006] Antibodies sometimes denature due to physical stimuli such as heating and vibration, interactions with compounds such as surfactants, oxidizing and reducing agents, and sugars, or during long-term storage, resulting in aggregation, decomposition, and chemical reactions. If the antibody denatures, the binding properties to antigens and Fc receptors change, so there are concerns that the function and effect of the antibody may be weakened, or that the aggregated antibody may cause inflammation. The ease of denaturation such as aggregability varies depending on the molecular species of the antibody.

[0007] Antibodies contain charged amino acids, highly polar amino acids, and sugar chains. Therefore, there is a situation where an uneven distribution of ions occurs due to interaction with ions in the antibody solution, causing a change in the pH of the antibody solution (pH drift) (Donnan effect). The change in the pH of the antibody solution sometimes affects the storage stability of the antibody, and the strength of the Donnan effect varies depending on the molecular species of the antibody.

[0008] High-concentration antibody solutions tend to form solutions with high self-viscosity due to the nature 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 due to an increase in the antibody concentration varies depending on the molecular species of the antibody.

[0009] In addition, when the antibody solution is stored for a long time, deterioration phenomena such as changes in pH, formation of insoluble and / or soluble aggregates become problems, but these also vary depending on the molecular species of the antibody.

[0010] Generally, for pharmaceutical compositions containing antibodies, various efforts have been made to produce stabilized pharmaceutical compositions with little loss of the active ingredient even after long-term storage. The antibody as an active ingredient and various additives such as buffers are dissolved and manufactured in the form of a pharmaceutical composition. However, especially in pharmaceutical compositions containing a high concentration of antibodies, the technology for preventing antibody aggregation, turbidity, viscosity increase, and pH drift is still insufficient.

[0011] The inventors of the present application have obtained multiple 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.

[0012] Prior Art Documents

[0013] Patent Documents

[0014] Patent Document 1: International Publication No. 2015 / 099175 Summary of the Invention

[0015] Problems to be Solved by the Invention

[0016] An object of the present invention is to provide a pharmaceutical composition containing a human anti-IL-33 monoclonal antibody that is suitable for administration to a subject.

[0017] Means for Solving the Problems

[0018] The inventors of the present application found that turbidity occurred when formulating a human anti-IL-33 monoclonal antibody preparation, and found that the cause of the turbidity was the formation of aggregates. In addition, it was found that the turbidity depended on the NaCl concentration and pH. Based on the above insights, the present invention was thus completed.

[0019] That is, the present invention provides the following.

[0020] [1] A pharmaceutical composition containing a human anti-IL-33 monoclonal antibody as an active ingredient, wherein

[0021] the combination of the amino acid sequences of the complementarity-determining region 1 (H1) of the heavy chain, the complementarity-determining region 2 (H2) of the heavy chain, the complementarity-determining region 3 (H3) of the heavy chain, the complementarity-determining region 1 (L1) of the light chain, the complementarity-determining region 2 (L2) of the light chain, and the complementarity-determining region 3 (L3) of the light chain of the human anti-IL-33 monoclonal antibody is any one of C1 to C5 in Table 1,

[0022] the pharmaceutical composition substantially does not contain sodium chloride, or contains less than 30 mM of sodium chloride.

[0023] [Table 1]

[0024] In Table 1, the following sequence numbers represent the sequence numbers in the sequence listing.

[0025] H1 H2 H3 L1 L2 L3 C1 Serial number 11 Serial number 12 Serial number 13 Serial number 14 Serial number 15 Serial number 16 C2 Serial number 17 Serial number 18 Serial number 19 Serial number 20 Serial number 21 Serial number 22 C3 Serial number 17 Serial number 23 Serial number 24 Serial number 25 Serial number 26 Serial number 27 C4 Serial number 28 Serial number 29 Serial number 30 Serial number 25 Serial number 31 Serial number 32 C5 Serial number 17 Serial number 33 Serial number 34 Serial number 35 Serial number 36 Serial number 37

[0026] [2] The pharmaceutical composition according to [1], wherein the combination of the 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.

[0027] [Table 2]

[0028] In Table 2, the following sequence numbers represent the sequence numbers in the sequence listing.

[0029] Heavy chain variable region Light chain variable region V1 Serial number 38 Serial number 39 V2 Serial number 40 Serial number 41 V3 Serial number 42 Serial number 43 V4 Serial number 44 Serial number 45 V5 Serial number 46 Serial number 47

[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 substantially does not 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 be 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 of acetate, histidine or phosphate.

[0036] [9] The pharmaceutical composition according to any one of [1] to [8], wherein the pH is adjusted using 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] , which contains 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 according to any one of [1] to [15-1], the pharmaceutical composition contains 10 mM histidine, 4% (w / v) sorbitol, 0.02% (w / v) polysorbate 80, 150 mg / ml of the active ingredient, and the pH is adjusted to 5.5 to 6.5.

[0046]

[17] The pharmaceutical composition as described in

[16] , which is for subcutaneous administration.

[0047]

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

[0048]

[19] The pharmaceutical composition as described in

[18] , which is for intravenous administration.

[0049]

[20] The pharmaceutical composition as described in any one of [1] to

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

[0050]

[21] The lyophilized preparation of the pharmaceutical composition as described in any one of [1] to

[20] .

[0051]

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

[0052] it includes administering a pharmaceutical composition containing a human anti-IL-33 monoclonal antibody as an active ingredient to a subject in need,

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

[0054] wherein the pharmaceutical composition substantially does not contain sodium chloride, or contains less than 30 mM of sodium chloride.

[0055]

[23] The method as described in

[22] , wherein the combination of the 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 as described in

[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 as described in 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 substantially does not contain 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 be 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 a buffer of acetate, histidine or phosphate.

[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 according to

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

[0069]

[37] The method according to any one of

[22] to

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

[0070]

[38] The method according to

[37] , wherein the administration is subcutaneous administration.

[0071]

[39] The method according to 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 an active ingredient at 10 mg / ml, and the pH is adjusted to 5.5 to 6.5.

[0072]

[40] The method according to

[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 IL-33 related diseases, characterized in that

[0075] it contains a human anti-IL-33 monoclonal antibody as an active ingredient,

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

[0077] wherein the pharmaceutical composition substantially does not contain sodium chloride, or contains less than 30 mM of sodium chloride.

[0078]

[43] Use for manufacturing a pharmaceutical composition for the treatment or prevention of IL-33 related diseases,

[0079] the pharmaceutical composition contains a human anti-IL-33 monoclonal antibody, substantially does not contain sodium chloride, or contains less than 30 mM of sodium chloride,

[0080] wherein the combination of the amino acid sequences of the complementarity determining region 1 (H1) of the heavy chain, the complementarity determining region 2 (H2) of the heavy chain, the complementarity determining region 3 (H3) of the heavy chain, the complementarity determining region 1 (L1) of the light chain, the complementarity determining region 2 (L2) of the light chain, and the complementarity determining region 3 (L3) of the light chain 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 sugar selected from the group consisting of sucrose and trehalose, and the sugar alcohol is a sugar selected from the group consisting of sorbitol and mannitol.

[0082]

[45] The pharmaceutical composition as described in [12-1] or

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

[0083]

[46] The pharmaceutical composition as described in any one of [1] to

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

[0084]

[47] The pharmaceutical composition as described in

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

[0085]

[48] The pharmaceutical composition as described in

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

[0086]

[49] The pharmaceutical composition as described in

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

[0087]

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

[0088] the combination of the amino acid sequences of the complementarity determining region 1 (H1), complementarity determining region 2 (H2), complementarity determining region 3 (H3) of the heavy chain, complementarity determining region 1 (L1), complementarity determining region 2 (L2) and complementarity determining region 3 (L3) of the light chain 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 as described in

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

[0091]

[52] The pharmaceutical composition as described in

[50] or

[51] , which substantially does not contain sodium chloride or contains less than 30 mM of sodium chloride.

[0092] Advantages of the Invention

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

[0094] Figure 1 Figure 1 Shows the turbidity of the P7N formulation of the A10-1C04 antibody.

[0095] Figure 2 Figure 2 Shows the turbidity inhibition effect brought about by the addition of sugar. In the formulations of the A10-1C04 antibody added with sorbitol (A5S, H6S, P6S, P7S), the turbidity was improved. DETAILED DESCRIPTION OF THE INVENTION

[0096] For ease of understanding of the present invention, the terms used in the present invention will be described below.

[0097] [Pharmaceutical composition]

[0098] In the present specification, a pharmaceutical composition refers to a composition prepared in a manner 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] [Substantially]

[0100] In the present specification, "substantially free of sodium chloride" means a case where sodium chloride is not added to the pharmaceutical composition of the present invention, etc., and indicates that the pharmaceutical composition of the present invention does not contain sodium chloride at a level that causes turbidity.

[0101] [Surfactant]

[0102] In the present specification, a surfactant refers to a general term for substances having a part that is easily hydrophilic (hydrophilic group) and a part that is easily lipophilic (lipophilic group · hydrophobic group) 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] ​​​​As used in this specification, the term "antibody" is used in its broadest sense and includes 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., bispecific antibody), a drug conjugate antibody (ADC), and can also be antigen-binding fragments such as dAbs, scFv, Fab, F(ab)'2, Fab'.

[0105] [Monoclonal antibody]

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

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

[0108] The 5 human anti-IL-33 monoclonal antibodies used in the present invention refer to the human anti-IL-33 monoclonal antibodies of 5 clones, namely A10-1C04, A23-1A05, A25-2C02, A25-3H04, and A26-1F02, which are disclosed in International Publication No. 2015 / 099175, and are human anti-IL-33 monoclonal antibodies composed of heavy chains and light chains of 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, a Complementary Determining Region (CDR) refers to the amino acid residues of an antibody that are related to antigen binding. CDRs are often also referred to as "hypervariable regions" and are the amino acid sequences specific to the molecular species of an antibody, which can be determined by methods such as those 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)). There are 3 CDRs in the light chain of an antibody (L1, L2, L3) and 3 in the heavy chain of an antibody (H1, H2, H3). The CDRs of the 5 clones, A10-1C04, A23-1A05, A25-2C02, A25-3H04, and A26-1F02, are as follows.

[0136] (a) A10-1C04:

[0137] H1: DYYMN (SEQ ID NO: 11 in the Sequence Listing)

[0138] H2: SISRYSSYIYYADSVKG (SEQ ID NO: 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 (SEQ ID NO: 15 in the Sequence Listing)

[0142] L3: QTYDSSRWV (SEQ ID NO: 16 in the Sequence Listing)

[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 (SEQ ID NO: 20 in the Sequence Listing)

[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 (Sequence Listing Serial Number 17)

[0152] H2: SISARSSYIYYADSVKG (Sequence Listing Serial Number 23)

[0153] H3: LATRNNAFDI (Sequence Listing Serial Number 24)

[0154] L1: SGSSSNIGRNAVN (Sequence Listing Serial Number 25)

[0155] L2: ASNMRVS (Sequence Listing Serial Number 26)

[0156] L3: WAWDDSQKVGV (Sequence Listing Serial Number 27)

[0157] (d) A25-3H04:

[0158] H1: RYYMH (Sequence Listing Serial Number 28)

[0159] H2: SISAQSSHIYYADSVEG (Sequence Listing Serial Number 29)

[0160] H3: LATRQNAFDI (Sequence Listing Serial Number 30)

[0161] L1: SGSSSNIGRNAVN (Sequence Listing Serial Number 25)

[0162] L2: ASNMRRS (Sequence Listing Serial Number 31)

[0163] L3: SAWDDSQKVVV (Sequence Listing Serial Number 32)

[0164] (e) A26-1F02:

[0165] H1: NYYMH (Sequence Listing Serial Number 17)

[0166] H2: SISARSSYLYYADSVKG (Sequence Listing Serial Number 33)

[0167] H3: LATRHVAFDI (Sequence Listing Serial Number 34)

[0168] L1: SGSSSNIGNNAVN (Sequence Listing Serial Number 35)

[0169] L2: ASNMRRP (Sequence Listing Serial Number 36)

[0170] L3: EAWDDSQKAVV (Sequence Listing Serial Number 37)

[0171] [Variable Region]

[0172] In this specification, the variable region refers to the part other than the constant region of a monoclonal antibody. It is related to the binding to an antigen and is the part that determines the specificity of the antibody that varies according to the antigen. In the variable region, there are a heavy chain variable region and a light chain variable region. The heavy chain variable regions and light chain variable regions of the 5 clones A10-1C04, A23-1A05, A25-2C02, A25-3H04, and A26-1F02 are as follows.

[0173] (a) A10-1C04:

[0174] · Heavy chain variable region:

[0175] EVQLLESGGGLVQPGGSLRLSCAASGFTFSDYYMNWVRQAPGKGLEWVSSISRYSSYIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDIGGMDVWGQGTLVTVSS (Sequence Listing Serial Number 38)

[0176] · Light chain variable region:

[0177] QSVLTQPPSASGTPGQRVTISCTGSSSNIGAVYDVHWYQQLPGTAPKLLIYRNNQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCQTYDSSRWVFGGGTKLTVLG (Sequence Listing Serial Number 39)

[0178] (b) A23-1A05:

[0179] · Heavy chain variable region:

[0180] EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMHWVRQAPGKGLEWVSSISARSRYHYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARLATRHNAFDIWGQGTLVTVSS (Sequence Listing Serial Number 40)

[0181] · Light chain variable region:

[0182] QSVLTQPPSASGTPGQRVTISCSGSSSNIGNNAVSWYQQLPGTAPKLLIYASNMRVIGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDDSQKALVFGGGTKLTVLG (SEQ ID NO: 41)

[0183] (c) A25-2C02:

[0184] · Heavy chain variable region:

[0185] EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMHWVRQAPGKGLEWVSSISARSSYIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARLATRNNAFDIWGQGTLVTVSS (SEQ ID NO: 42)

[0186] · Light chain variable region:

[0187] QSVLTQPPSASGTPGQRVTISCSGSSSNIGRNAVNWYQQLPGTAPKLLIYASNMRVSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCWAWDDSQKVGVFGGGTKLTVLG (SEQ ID NO: 43)

[0188] (d) A25-3H04:

[0189] · Heavy chain variable region:

[0190] EVQLLESGGGLVQPGGSLRLSCAASGFTFSRYYMHWVRQAP GKGLEWVSSISAQSSHIYYADSVEGRFTISRDNSKNTLYLQMNSLR AEDTAVYYCARLATRQNAFDIWGQGTLVTVSS (SEQ ID NO: 44)

[0191] · Light chain variable region:

[0192] QSVLTQPPSASGTPGQRVTISCSGSSSNIGRNAVNWYQQLPGT APKLLIYASNMRRSGVPDRFSGSKSGTSASLAISGLRSEDEADYYC SAWDDSQKVVVFGGGTKLTVLG (SEQ ID NO: 45)

[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 in the Sequence Listing)

[0198] "Stable"

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

[0200] "effector function"

[0201] The "effector function" of an antibody refers to those biological activities resulting from the Fc region of the antibody (the Fc region of the native sequence or the Fc region of an 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] [Cloudiness]

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

[0204] [Lyophilized preparation]

[0205] In the present specification, a "lyophilized preparation" refers to an almost water-free dried (e.g., lyophilized) pharmaceutical composition. The lyophilization technique of antibodies 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 and is also known as NF-HEV. IL-33 has the following functions: when released extracellularly as a cytokine, it binds to the IL-33 receptor (ST2 and IL-1RAcP), and initiates intracellular signal transduction in cells expressing the IL-33 receptor. The signal transduction induced by IL-33 is not restricted, and there are NF-κB pathway and MAPKKs pathway, ultimately leading to the production of various cytokines, chemokines, and inflammatory mediators. Examples of cytokines induced by IL-33 include TNF-α, IL-1β, IL-3, IL-4, IL-5, IL-6, IL-13, etc., and in particular, IL-5, IL-6, and IL-13 can be induced. Examples of chemokines induced by IL-33 include CXCL2, CCL2, CCL3, CCL6, CCL17, CCL24, etc. Examples of inflammatory mediators induced by IL-33 include PGD2, LTB4, etc. The cytokines, chemokines, and inflammatory mediators induced by IL-33 are involved 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 to inhibit at least one of the aforementioned functions, it can refer to either full-length IL-33 or mature IL-33, and can also be a derivative or mutant having homology with them. Additionally, it can also be human IL-33 or IL-33 derived from other organisms.

[0208] [Pharmaceutically acceptable]

[0209] In the present specification, the term "pharmaceutically acceptable" means that it does not interfere with the effectiveness of the active ingredient(s) (which can be plural kinds) and is non-toxic.

[0210] [Isotonic]

[0211] When used in the present specification, 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 based on 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 treatments 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 explaining the present invention, and the present invention is not limited to these embodiments.

[0215] The human anti-IL-33 monoclonal antibody involved in the present invention can be produced by known techniques, such as the method described in Patent Document 1.

[0216] The monoclonal antibody produced as described above can be formulated into a desired composition by methods such as dialysis, ultrafiltration devices, or ammonium sulfate precipitation. Additionally, after preparing a solution containing the desired buffer, sugars, surfactants, etc. can be added later for formulation.

[0217] The inventors of the present application found that when five human anti-IL-33 monoclonal antibodies are made into a solution, partial aggregation occurs and turbidity appears. The turbidity is caused by the formation of fine particles as aggregates containing the antibody. When formulating a pharmaceutical composition, there are the following concerns: the biological activity of the antibody as the active ingredient decreases, the pharmacokinetics deteriorates due to the induction of anti-drug antibodies (ADA) by aggregates with high immunogenicity, and inflammation is caused by the aggregates themselves. Therefore, it is necessary to suppress turbidity. Thus, the present invention relates to a pharmaceutical composition with suppressed turbidity. The pharmaceutical composition of the present invention can contain five human anti-IL-33 monoclonal antibodies as the active ingredient, and can also contain salts, buffers, surfactants, sugars, etc.

[0218] [Salt concentration]

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

[0220] [Buffer and pH]

[0221] The pharmaceutical composition of the present invention uses a buffer to adjust the pH. As a buffer suitable for the pharmaceutical composition, there is no limitation, and examples thereof include gluconate, histidine, citrate, phosphate [e.g., sodium or potassium], succinate [e.g., sodium], acetate, tris(hydroxymethyl)aminomethane, glycine, arginine, and combinations thereof, and they 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 only required to be pharmaceutically acceptable, and there is no particular limitation. It is preferably 1 mM to 150 mM, more preferably 5 mM to 100 mM, and further preferably 10 mM to 50 mM. In addition, the concentration of the buffer is preferably about 1 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, or about 50 mM. From the viewpoint of using a buffer of salt, especially inorganic salt, a concentration of less than 30 mM, especially 10 mM or less, is preferably used.

[0222] In the pharmaceutical composition of the present invention, as a buffer, histidine (e.g., at a concentration of 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 beneficial. In one embodiment, the stable pharmaceutical composition contains 5 mM to 20 mM of histidine. The pH of the pharmaceutical composition can be in the range of 4.0 to 8.0, a pH in the range of 4.5 to 7.5, such as 5.0 to 7.0, 5.2 to 6.8, e.g., 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 pH of the pharmaceutical composition of the present invention is 4 or 8, the turbidity increases during long-term storage. In addition, when the pH of the pharmaceutical composition of the present invention is 4 or 8, the pH drift is significant during long-term storage. Therefore, in one embodiment, the pH of the stable antibody-containing pharmaceutical composition is greater than 4 and less than 8, preferably 5 or more and 7 or less, more preferably 5.5 or more and 6.5 or less, and most preferably 6.0.

[0223] [Surfactant]

[0224] The pharmaceutical composition of the present invention preferably contains a surfactant. There is no limitation on the surfactant suitable for the pharmaceutical composition, including nonionic surfactants, ionic surfactants, zwitterionic surfactants, and combinations thereof. There is no limitation on the common surfactant used in the present invention, including sorbitan fatty acid esters (e.g., sorbitan monooctanoate, sorbitan monolaurate, sorbitan monopalmitate), sorbitan trioleate, glycerol fatty acid esters (e.g., glycerol monocaprylate, glycerol monomyristate, glycerol monostearate), polyglycerol fatty acid esters (e.g., decaglycerol monostearate, decaglycerol distearate, decaglycerol monooleate), polyoxyethylene sorbitan fatty acid esters (e.g., polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monooleate, polyoxyethylene sorbitan monostearate, polyoxyethylene sorbitan monopalmitate, polyoxyethylene sorbitan trioleate, polyoxyethylene sorbitan tristearate), polyoxyethylene sorbitol fatty acid esters (e.g., polyoxyethylene sorbitol tetrastearate, polyoxyethylene sorbitol 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., polyoxyethylene lauryl ether), polyoxyethylene polyoxypropylene alkyl ethers (e.g., polyoxyethylene polyoxypropylene glycol, polyoxyethylene polyoxypropylene propyl ether, polyoxyethylene polyoxypropylene cetyl ether), polyoxyethylene alkyl phenyl ethers (e.g., polyoxyethylene nonyl phenyl ether), polyoxyethylene hydrogenated castor oil (e.g., polyoxyethylene castor oil, polyoxyethylene hydrogenated castor oil), polyoxyethylene beeswax derivatives (e.g., polyoxyethylene sorbitol beeswax), polyoxyethylene lanolin derivatives (e.g., polyoxyethylene lanolin), polyoxyethylene fatty acid amides (e.g., polyoxyethylene stearamide), 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 sulfosuccinates (e.g., sodium lauryl sulfosuccinate), and natural surfactants such as lecithin, glycerophospholipids, sphingomyelin phospholipids (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 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 alkyl phenyl 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 can be any concentration commonly used in the art. As an example, it can be used at a concentration of about 0.01% (w / v) to about 0.1% (w / v), such as about 0.01% (w / v) to about 0.04% (w / v), such as 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), about 0.1% (w / v). Among the surfactants, especially polysorbate 80 (Tween 80) is particularly beneficial. In one embodiment, the stable pharmaceutical composition contains about 0.02% (w / v) polysorbate 80. In one embodiment, the stable pharmaceutical composition contains about 0.02% (w / v) polysorbate 20.

[0226] [Polyol]

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

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

[0229] Polyols are also called polyalcohols, and any molecule having two or more alcoholic hydroxyl groups can be used. For example, glycerol, propylene glycol, polyethylene glycol (PEG), saccharides, etc. can be mentioned, and saccharides can be preferably selected.

[0230] The saccharides suitable for the pharmaceutical composition of the present invention are not limited and include compounds having the general formula (CH2O)n and their derivatives such as monosaccharides, disaccharides, trisaccharides, polysaccharides, sugar alcohols, reducing sugars, non-reducing sugars, etc. Examples of saccharides include, as monosaccharides, glucose, fructose, and galactose; as disaccharides, sucrose, trehalose, lactose, maltose, lactulose, maltulose, isomaltulose, and melibiose; as trisaccharides, melezitose, raffinose, and maltotriose; as polysaccharides, stachyose and dextran; as sugar alcohols, sorbitol, mannitol, erythritol, maltitol, lactitol, arabinitol, xylitol, etc. Among monosaccharides, disaccharides, and trisaccharides, as reducing sugars, glucose, fructose, lactose, maltose, lactulose, maltulose, isomaltulose, melibiose, melezitose, and maltotriose can be cited; as non-reducing sugars, trehalose, sucrose, and raffinose can be cited. As the saccharide 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 acceptable, and is preferably 50 mM to 300 mM, more preferably 165 mM to 275 mM, and further preferably 200 mM to 220 mM. In addition, the concentration of the saccharide is preferably about 50 mM, about 55 mM, about 100 mM, about 110 mM, about 150 mM, about 165 mM, about 200 mM, about 220 mM, about 275 mM, about 300 mM. In one embodiment, the stable pharmaceutical composition contains 3% to 5% (w / v) (165 to 275 mM) of sorbitol. In one embodiment, the stable pharmaceutical composition contains about 3.6% (w / v) or about 4% (w / v) (about 200 mM or about 220 mM, 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 polyols.

[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, and 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. When the antibody concentration of the pharmaceutical composition of the present invention is 175 mg / mL or more, the viscosity is high, so it is preferably less than 175 mg / mL, and more preferably 150 mg / mL or less. 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 an appropriate viscosity, and can be easily administered to any patient. The lower the viscosity, the easier it is to administer without force. Regarding the pharmaceutical composition of the present invention, the inventors of the present application found that when the viscosity is about 20 cP or more, it is slightly difficult to eject from the syringe at the time of administration. Therefore, the viscosity of the pharmaceutical composition of the present invention is preferably 50 cP or less, more preferably 30 cP or less, further preferably 20 cP or less, and most preferably 10 cP or less. In one embodiment, the pharmaceutical composition of the present invention is about 1, about 2, about 5, about 10, about 15, about 20 cP. The viscosity of the present invention can be measured using a rheometer (rotary 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), endometriosis, etc. The administration method is not particularly limited and can be systemic administration or local administration. 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, for example, the dose is limited, so it preferably contains a high concentration of human anti-IL-33 antibody. For example, it is preferably a pharmaceutical composition containing 10 mM histidine, 4% (w / v) sorbitol, 0.02% (w / v) polysorbate 80, 150 mg / ml of the active ingredient, and the pH is adjusted to 5.5 to 6.5. When the pharmaceutical composition of the present invention is administered intravenously, for example, it can contain a low concentration of human anti-IL-33 antibody. For example, it is preferably a pharmaceutical composition containing 10 mM histidine, 3.6% (w / v) sorbitol, 0.02% (w / v) polysorbate 80, 10 mg / ml of the active ingredient, and the pH is adjusted to 5.5 to 6.5.

[0238] As examples of IL-33-related diseases, without limitation, there can be cited asthma, atopic dermatitis, urticaria, pollinosis, 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 diseases (e.g., ulcerative colitis, Crohn's disease, gluten-sensitive enteropathy, etc.), systemic lupus erythematosus, Sjogren's syndrome, Behcet's disease, pemphigus, pemphigoid, autoimmune hemolytic anemia, autoimmune inflammatory eye diseases, autoimmune neonatal thrombocytopenia, autoimmune neutropenia, autoimmune oophoritis and orchitis, autoimmune thrombocytopenia, autoimmune thyroiditis, polymyositis, dermatomyositis, myasthenia gravis, adrenaline agonist tolerance, alopecia greata, antiphospholipid syndrome, autoimmune diseases of the adrenal gland (e.g., autoimmune Addison's disease, etc.), celiac-dermatitis, chronic fatigue immune dysfunction syndrome (CFIDS), cold agglutinin disease, idiopathic mixed cryoglobulinemia, fibromyalgia-fibromyositis, glomerulonephritis (e.g., IgA nephropathy, etc.), Graves' disease, hyperthyroidism (i.e., Hashimoto's thyroiditis), idiopathic thrombocytopenic purpura (ITP), mixed connective tissue disease, type 1 or immune-mediated diabetes, pernicious anemia, polychrondritis, polyglandular syndrome, general myotonia syndrome, vitiligo, sarcoidosis, polyendocrinopathy syndrome, other endocrine gland insufficiencies, 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 tumors, laryngeal cancer, lip and oral cavity cancer, hypopharyngeal cancer, thyroid cancer, esophageal cancer, breast cancer, lung cancer, gastric cancer, adrenocortical cancer, cholangiocarcinoma, gallbladder cancer, liver cancer, pancreatic cancer, bladder cancer, colorectal cancer, uterine cancer, ovarian cancer, prostate cancer, testicular cancer, chronic lymphocytic leukemia, chronic myeloid leukemia, Ewing sarcoma, Hodgkin lymphoma, non-Hodgkin lymphoma, melanoma, mesothelioma, multiple myeloma, etc.), infections resistant to rejection from the immune system (e.g., severe acute respiratory syndrome (SARS)), lethal cytokine storms associated with highly virulent influenza infections, and scurvy. 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. can be preferably selected.

[0239] The "stable" pharmaceutical composition of the present invention shows no significant changes at least for 12 months, preferably 2 years, more preferably 3 years at refrigerated temperature (2 - 8°C), or at least for 3 months, preferably 6 months, more preferably 1 year at room temperature (22 - 28°C). For example, after storing at 5°C for 2 years, there is no turbidity, the OD650 is 0.014 or less, preferably 0.01 or less, more preferably 0.008 or less, the pH drift is 1 or less, preferably 0.8 or less, more preferably 0.5 or less, the 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 particles / mL or less, preferably 1000 particles / mL or less, more preferably 750 particles / mL or less, most preferably 500 particles / mL or less.

[0240] The pharmaceutical composition of the present invention can be further appropriately added with preservatives, anti-adsorbents, analgesics, sulfur-containing reducing agents, antioxidants, etc. as needed.

[0241] As the preservative, there is no particular limitation as long as it is pharmaceutically acceptable. For example, methyl paraben, ethyl paraben, sorbic acid, phenol, cresol, and chlorocresol can be cited.

[0242] As the anti-adsorbent, there is no particular limitation as long as it is pharmaceutically acceptable. For example, human serum albumin, lecithin, dextran, ethylene oxide - propylene oxide copolymer, hydroxypropyl cellulose, methyl cellulose, polyoxyethylene hydrogenated castor oil, and polyethylene glycol can be cited.

[0243] As the analgesic, there is no particular limitation as long as it is pharmaceutically acceptable. For example, local anesthetics such as lidocaine can be cited.

[0244] As the sulfur-containing reducing agent, there is no particular limitation as long as it is pharmaceutically acceptable. For example, mention may be made of substances having a mercapto 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] As the antioxidant, there is no particular limitation as long as it is pharmaceutically acceptable. For example, mention may be made of isoascorbic acid, dibutylhydroxytoluene, butylhydroxyanisole, α-tocopherol, tocopherol acetate, L-ascorbic acid and its salts, L-ascorbic acid palmitate, L-ascorbic acid stearate, sodium bisulfite, sodium sulfite, tripentyl gallate, propyl gallate, or chelating agents such as disodium ethylenediaminetetraacetate (EDTA), sodium pyrophosphate, and sodium metaphosphate.

[0246] For the pharmaceutical composition of the present invention, in order to preserve it for a longer period, it may also be a freeze-dried preparation.

[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] [Examples]

[0249] Comparative Example 1: Turbidity of a pharmaceutical composition containing a 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 to have a concentration of 150 mg / mL in a solvent (10 mM sodium phosphate / pH 7 / 150 mM NaCl / 0.02% (w / v) polysorbate 80 (Polysobate80)) (hereinafter simply referred to as "P7N"). Based on visual confirmation of the properties, turbidity was observed as a result (for example, regarding A10-1C04, shown in Figure 1 ). It should be noted that the presence or absence of turbidity was confirmed by visual observation under white light (13W fluorescent lamp) against a black background. In addition, for this pharmaceutical composition, insoluble particles (particles of 5 μm or more) were measured using FlowCam (manufactured by FluidImaging Technologies), and as a result, 1,919 particles / mL were detected. Based on the above results, it was considered that the human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02) had high cohesiveness and were 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] To eliminate the turbidity of human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02), the buffer was changed as described below based on the formulation of Comparative Example 1, thereby changing the pH of the pharmaceutical composition. The antibody concentration was set at 150 mg / ml. The studied formulations are shown below.

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

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

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

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

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

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

[0259] The evaluation of the properties was carried out. As a result, turbidity occurred in all the formulations with pH 4, 5, 6, 7, and 8. Based on this result, it was considered that even if the pH of the formulation was changed, the turbidity would not be improved.

[0260] It should be noted that the visual confirmation of the properties was observed by the method described in Comparative Example 1.

[0261] Example 1: Turbidity inhibition effect of added sugar

[0262] To eliminate the turbidity of human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02), the following formulations obtained by adding 5% sorbitol and replacing the sodium chloride contained in A4N, A5N, H6N, P6N, P7N, and P8N of the formulation of Comparative Example 2 were evaluated. The antibody concentration was set at 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 evaluation of the properties and OD650 (turbidity) was carried out. As a result, by adding sorbitol instead of sodium chloride, the white turbidity was improved (for example, regarding A10-1C04, shown in Figure 2 ). In addition, by adding sorbitol, even after 3 months at 40 °C, the increase in OD650 was suppressed, and the suppression effect was particularly good at pH 5-7 (Table 3).

[0270] It should be noted that the confirmation of the properties by the naked eye was observed by the method described in Comparative Example 1. For OD650, for 100 μL of the solution of each formulation, the absorbance at 650 nm was measured using a Molecular Device microplate reader (SoftMaxPro software).

[0271] [Table 3]

[0272] Table 3: OD650 of various formulations

[0273] Formulation During preparation After storage at 40°C for 3 months 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), which is an index of aggregation, was measured when human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04, or A26-1F02) were dissolved in a buffer of 10 mM histidine / pH 6.0 or pH 5.5 and 0, 50, or 100 mM of NaCl was added. The measurement temperature was set at 25°C, and the antibody concentration was prepared to be 0.5, 1, 2.5, 5, 10, or 20 mg / mL. As a result, when the NaCl addition amount was 50 mM or more, the Kd value became negative (aggregation increased), so it was considered that the NaCl addition amount 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 the dynamic light scattering method, the slope is obtained from the coordinate graph of the antibody concentration (horizontal axis) and the diffusion coefficient (vertical axis), and this slope is divided by the diffusion coefficient (D0) at a concentration of 0. That is, the Kd value is calculated from the following relational expression. When the value of the Kd value is positive and the larger it is, the lower the aggregation.

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

[0277] [Table 4]

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

[0279] Salt concentration pH 5.5 pH 6 0 mM 19.2 0.17 50 mM -4.7 -6.2 100 mM -6.4 -7.6

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

[0281] 0, 5, 10, or 30 mM of NaCl was added to 10 mM histidine / pH 6.0 / 3.6% (w / v) sorbitol, and the Kd value, which is an index of aggregation, was measured. A10-1C04 at 2.5, 5, 10, or 14 mg / mL was used as the antibody. As a result, when the NaCl addition amount was 30 mM or more, the Kd value became negative (the Kd values for the addition of 0, 5, 10, and 30 mM of NaCl were 17.3, 7.3, 1.2, and -4.0 mL / g, respectively), so it was considered that the NaCl addition amount 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 change in pH during long-term storage of a pharmaceutical composition containing a human anti-IL-33 monoclonal antibody (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02) was measured. The antibody concentration was set at 150 mg / mL, and formulations with pH values ranging from 4 to 8 were investigated (A4S, A5S, H6S, P6S, P7S, P8S). For each formulation, the pH was measured at the time of preparation and after storage at 40 °C for 2, 4, 8, and 12 weeks. As a result, significant pH drift was observed in the formulations with pH 4 (A4S) and pH 8 (P8S) (for example, for A10-1C04, shown in Table 5). Therefore, formulations with pH values of 5 to 7, which exhibit less pH drift, are considered to be better.

[0284] [Table 5]

[0285] Table 5: pH drift during storage at 40 °C

[0286] Formulation 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 such that the antibody concentration of a human anti-IL-33 monoclonal antibody (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02) in a formulation of 10 mM histidine / pH 6 / 3.6% (w / v) sorbitol / 0.02% (w / v) polysorbate 80 was 15, 50, 100, 125, 150, 175, or 200 mg / ml. The viscosity at a measurement temperature of 25 °C was measured using a viscometer (Brookfield DV3TLVCJ0 type viscometer (CPA-40Z rotor, CPA-44YZ sample cup)). The viscosities of A10-1C04 at antibody concentrations of 15, 50, 100, 125, 150, 175, and 200 mg / mL were 1.19, 1.84, 5.33, 9.15, 16.29, 47.98, and 84.96 cP, respectively, indicating that the viscosity increased sharply after exceeding 150 mg / mL. Additionally, solutions with various viscosities were prepared, and the injectability using a syringe was investigated. As a result, it was found that administration became slightly difficult when the viscosity exceeded approximately 20 cP, suggesting that the antibody concentration should be less than 175 mg / ml.

[0289] Example 6: Effect of sugar addition on aggregation evaluation and inhibition of particle generation

[0290] 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 in a 10 mM histidine / pH 6.0 composition, and the effects on the Kd value as an aggregation index were confirmed in the presence of added sucrose or sorbitol. The Kd value was calculated by the method described in Example 2. For example, for the Kd value in the case of using A10-1C04, it was 33.4 mL / g without added sugar, and 24.7 and 33.5 mL / g, respectively, when 3.6% (w / v) sucrose or sorbitol was added, both being positive values. Based on this result, it was considered that sucrose could also be added instead of sorbitol. In addition, for a formulation with an antibody concentration of 10 mg / ml and 3.6% (w / v) sorbitol added (10 mM histidine / pH 6.0 / 3.6% (w / v) sorbitol), microparticles of 1.5 μm or more were measured at the time of preparation and after storage at 50 °C for 1 week using a particle counter HIAC (HACH System 9703+ type). For the measurement using HIAC, each sample was measured 4 times with an injection volume of 100 μL, and the data from the first measurement was discarded. As a result, the increase in the number of microparticles was suppressed in the case of adding 3.6% (w / v) sorbitol compared to when no sorbitol was added, and thus it was considered that there was an aggregation inhibitory effect due to the addition of sorbitol (for example, for A10-1C04, shown in Table 6).

[0291] [Table 6]

[0292] Table 6: Inhibitory effect of sorbitol on microparticle formation (unit: number / 100 μL)

[0293]

[0294] Example 7: Aggregation inhibition effect of surfactant

[0295] A 10 mM histidine / pH 6 / 3.6% (w / v) sorbitol composition of 10 mg / mL human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02) was prepared, and the effects on aggregation were confirmed in the presence of added 0.02% (w / v) polysorbate 20 or polysorbate 80 as a surfactant. No aggregation was confirmed at the time of preparation whether or not a surfactant was added.

[0296] Example 8: Stability of subcutaneous administration preparation and intravenous administration preparation

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

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

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

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

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

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

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

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

[0305] For example, the evaluation of the properties of 150 mg / mL of A10-1C04 was carried out. As a result, the turbidity was improved due to the absence of sodium chloride addition. 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] Formulation Properties A4 - A5 - H6 - H7 - P8 +

[0309] -: Clear (no turbidity)

[0310] +: Turbidity occurs

[0311] Example 10: Effect of salt on the turbidity of a pharmaceutical composition containing a human anti-IL-33 monoclonal antibody (2)

[0312] To eliminate the 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 of Example 9, namely A4, A5, and H6, and evaluations were conducted.

[0313] · 10 mM sodium acetate / pH 4 / 10 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter simply referred to as "A4N10")

[0314] · 10 mM sodium acetate / pH 4 / 30 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter simply referred to as "A4N30")

[0315] · 10 mM sodium acetate / pH 4 / 50 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter simply referred to as "A4N50")

[0316] · 10 mM sodium acetate / pH 4 / 100 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter simply referred to as "A4N100")

[0317] · 10 mM sodium acetate / pH 5 / 10 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter simply referred to as "A5N10")

[0318] · 10 mM sodium acetate / pH 5 / 30 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter simply referred to as "A5N30")

[0319] · 10 mM sodium acetate / pH 5 / 50 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter simply referred to as "A5N50")

[0320] · 10 mM sodium acetate / pH 5 / 100 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter simply referred to as "A5N100")

[0321] · 10 mM histidine / pH 6 / 10 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter simply referred to as "H6N10")

[0322] · 10 mM histidine / pH 6 / 30 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "H6N30")

[0323] · 10 mM histidine / pH 6 / 50 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "H6N50")

[0324] · 10 mM histidine / pH 6 / 100 mM NaCl / 0.02% (w / v) polysorbate 80 (hereinafter referred to as "H6N100")

[0325] For example, the properties of 150 mg / mL of A10-1C04 were evaluated. As a result, under the conditions of pH 4 to 6, turbidity was suppressed when sodium chloride was 10 mM, while turbidity was observed when sodium chloride was more than 30 mM (Table 8).

[0326] [Table 8]

[0327] Table 8: Properties of various formulations

[0328] None N10 N30 N50 N100 A4 - - + + + A5 - - + + + H6 - - + + +

[0329] -: Clear (no turbidity)

[0330] +: Turbidity occurred

[0331] Example 11: Aggregation inhibition effect of surfactant (2)

[0332] To confirm the effect of surfactants 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 containing surfactants were prepared. Then, these samples were rotated at 30 rpm in a thermostat at 50 °C using a spinner. The samples taken over time were visually inspected for 5 seconds against a white board and a black board as backgrounds under a fluorescent lamp of 2000 - 3750 Lux. In the system without added surfactant, a large amount of aggregates was observed after 2 days, while in the samples containing polysorbate 80 and poloxamer 188, no aggregates were observed even after 7 days. Especially in the system containing 0.02% or more of polysorbate 80, no aggregates were observed even after 14 days. In the samples containing polysorbate 20, no foreign substances were observed until 4 days, and although a small amount of foreign substances was observed after 7 days, no foreign substances were observed after 14 days. For example, the results for A10-1C04 are shown in Table 9.

[0333] 10 mM histidine / pH 6 (hereinafter referred to as "H6(-)")

[0334] 10 mM histidine / pH 6 / 0.02% (w / v) poloxamer 188 (hereinafter simply referred to as "H6PX")

[0335] 10 mM histidine / pH 6 / 0.02% (w / v) polysorbate 20 (hereinafter simply referred to as "H6P20")

[0336] 10 mM histidine / pH 6 / 0.01% (w / v) polysorbate 80 (hereinafter simply referred to as "H6PS1")

[0337] 10 mM histidine / pH 6 / 0.02% (w / v) polysorbate 80 (hereinafter simply referred to as "H6PS2")

[0338] 10 mM histidine / pH 6 / 0.05% (w / v) polysorbate 80 (hereinafter simply referred to as "H6PS5")

[0339] [Table 9]

[0340] Table 9: Aggregates observed in the specimens stored at 50 °C at 30 rpm

[0341] 0 days 1 day 2 days 3 days 7 days 14 days H6(-) - - ++ ++ ++ ++ H6PX - - - - - + H6P20 - - - -* + - H6PS1 - - - - - + H6PS2 - - - - - - H6PS5 - - - - - -

[0342] -: No aggregation

[0343] +: A small amount of aggregation was confirmed

[0344] ++: A large amount of aggregation was confirmed

[0345] *: Removed after 4 days

[0346] Example 12: Sugar addition and aggregation evaluation (2)

[0347] Prepare a 10 mM histidine / pH 6.0 composition of a 10 mg / mL human anti-IL-33 monoclonal antibody (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02), add mannitol, trehalose, sucrose or sorbitol to prepare a pharmaceutical solution. Then, freeze these specimens at -80 °C for 8 hours or more, and then allow them to thaw at room temperature for 4 hours or more. Repeat the above operation 6 times. Then, visually inspect for 5 seconds against a white board and a black board as the background under a fluorescent lamp of 2000 - 3750 Lux. By adding sugar, compared with the system without addition, the amount of aggregates after freeze-thawing becomes less, and especially in the systems using sorbitol and sucrose, an inhibitory effect on aggregates is shown. For example, the results for A10-1C04 are shown in Table 10.

[0348] 10 mM histidine / pH 6 (hereinafter simply referred to as "H6(-)")

[0349] 10 mM histidine / pH 6 / 3.0% (w / v) sorbitol (hereinafter referred to as "H6So3")

[0350] 10 mM histidine / pH 6 / 3.6% (w / v) sorbitol (hereinafter referred to as "H6So3.6")

[0351] 10 mM histidine / pH 6 / 4.0% (w / v) sorbitol (hereinafter referred to as "H6So4")

[0352] 10 mM histidine / pH 6 / 5.0% (w / v) sorbitol (hereinafter referred to as "H6So5")

[0353] 10 mM histidine / pH 6 / 3.6% (w / v) sucrose (hereinafter referred to as "H6Su")

[0354] 10 mM histidine / pH 6 / 3.6% (w / v) trehalose (hereinafter referred to as "H6Tr")

[0355] 10 mM histidine / pH 6 / 3.6% (w / v) mannitol (hereinafter referred to as "H6Ma")

[0356] [Table 10]

[0357] Table 10: Aggregates Observed by Visual Inspection after Freeze-Thaw

[0358]

[0359] -: No aggregation

[0360] +: A small amount of aggregation was confirmed

[0361] ++: A large amount of aggregation was confirmed

[0362] Example 13: Lyophilization

[0363] Prepare a composition of 10 mM histidine / pH 6.0 / 0.02% (w / v) polysorbate 80 of 150 mg / mL human anti-IL-33 monoclonal antibody (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02). After adding mannitol, trehalose, sucrose or sorbitol, freeze-drying was carried out by shelf freeze-drying (Kyowa Vacuum Co., Ltd.). Regarding the properties, the shape of the cake after freeze-drying was visually confirmed.

[0364] For redissolution, add water for injection and leave it to stand at 5°C for half a day, then confirm whether there are any lumps remaining in the solution. The lumps of any specimen were in good condition, and redissolution 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 the implementation for A10-1C04 are shown in Table 11.

[0365] 10 mM histidine / pH 6 / 0.02% (w / v) polysorbate 80 / 4.0% (w / v) sorbitol (hereinafter simply referred to as "LYSO")

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

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

[0368] 10 mM histidine / pH 6 / 0.02% (w / v) polysorbate 80 / 2.0% (w / v) mannitol (hereinafter simply referred to as "LYMA")

[0369] [Table 11]

[0370] Table 11: Physical properties after freeze-drying

[0371] LYSO LYSU LYTR LYMA Appearance Good Good Good Good Redissolution Sol Sol Sol Sol

[0372] Good: Lump formed, Poor: Lump not formed

[0373] Sol: Soluble, Dis: Insoluble

[0374] Example 14: Stability test (1)

[0375] Prepare a composition of 10 mM histidine / pH 6 / 0.02% (w / v) polysorbate 80 / 3.6% (w / v) sorbitol containing 10 mg / mL of human anti-IL-33 monoclonal antibody (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02), fill it into glass vials, seal with a halogenated butyl rubber stopper, and store it at 2 - 8 °C for 1 year, 2 years and 3 years as a long-term stability test. Visually inspect for 5 seconds against a white board and a black board background under a fluorescent lamp of 2000 - 3750 Lux to confirm the presence of aggregation. The binding activity was evaluated by the following method. Add human IL-33 to a 96-well plate and immobilize it overnight. After blocking with BSA, add the sample solution to each well, react with an HRP-labeled anti-human IgG antibody, and develop color using TMB. Then, measure the absorbance at 450 nm and 650 nm using a microplate reader (Molecular Device Corporation) to calculate the EC50. Calculate the EC50 of the standard solution measured in the same way and calculate the ratio. No aggregation or pH drift was observed at any measurement point, and the binding activity of the antibody did not decrease. For example, the results for A10-1C04 are shown in Table 12.

[0376] [Table 12]

[0377] Table 12: Long-term stability test (1)

[0378] 2～8℃ T0 2 - 8°C for 1 year 2 - 8°C for 2 years 2 - 8°C for 3 years Visual inspection - - - - Binding activity 100％ 110％ 101％ 101％ pH 6.0 6.0 6.0 6.0

[0379] -: No aggregation

[0380] +: A small amount of aggregation was confirmed

[0381] ++: A large amount of aggregation was confirmed

[0382] Example 15: Stability test (2)

[0383] Prepare a composition of 10 mM histidine / pH 6 / 0.02% (w / v) polysorbate 80 / 4.0% (w / v) sorbitol containing 150 mg / mL of human anti-IL-33 monoclonal antibody (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02), fill it into glass vials, seal with a halogenated butyl rubber stopper, and store it at 2 - 8 °C for 3 months and 6 months as a long-term stability test. Evaluate it by the method of Example 14. No aggregation or pH drift was observed at any measurement point, and the binding activity of the antibody did not decrease. For example, the results for A10-1C04 are shown in Table 13.

[0384] [Table 13]

[0385] Table 13: Long-term Stability Test (2)

[0386] 2～8℃ T0 2 - 8°C for 3 months 2 - 8°C for 6 months Visual inspection - - - Binding activity 115％ 110％ 100％ pH 5.9 5.8 5.8

[0387] -: No aggregation

[0388] +: A small amount of aggregation was confirmed

[0389] ++: A large amount of aggregation was confirmed.

Claims

1. A pharmaceutical composition comprising a human anti-IL-33 monoclonal antibody as an active ingredient, wherein, the combination of the amino acid sequences of the complementarity determining region 1 (H1), complementarity determining region 2 (H2), complementarity determining region 3 (H3) of the heavy chain, complementarity determining region 1 (L1), complementarity determining region 2 (L2), and complementarity determining region 3 (L3) of the light chain of the human anti-IL-33 monoclonal antibody is any one of C1 to C5 in Table 1, the pharmaceutical composition substantially does not contain sodium chloride, or contains less than 30 mM of sodium chloride, [Table 1] 。 2. The pharmaceutical composition according to claim 1, wherein the combination of the 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, [Table 2] 。 3. The pharmaceutical composition according to claim 1 or 2, wherein, the human anti-IL-33 monoclonal antibody is A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02.

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

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

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

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

8. The pharmaceutical composition according to any one of claims 1 to 7, wherein, The pH is adjusted using a buffer of acetate, histidine or phosphate.

9. The pharmaceutical composition according to any one of claims 1 to 8, wherein, The pH is adjusted using histidine.

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

11. The pharmaceutical composition according to any one of claims 1 to 10, wherein, The concentration of the active ingredient is 150 mg / ml or less.

12. The pharmaceutical composition according to any one of claims 1 to 11, which contains at least one polyol.

13. The pharmaceutical composition according to claim 12, wherein, The polyol is a sugar selected from the group consisting of disaccharides and sugar alcohols.

14. The pharmaceutical composition according to claim 12 or 13, wherein, The polyol is 3 to 5% (w / v) sorbitol.

15. The pharmaceutical composition according to any one of claims 1 to 14, which contains a surfactant.

16. The pharmaceutical composition according to claim 15, wherein, The surfactant is a non-ionic surfactant.

17. The pharmaceutical composition according to claim 16, wherein, The surfactant is polysorbate 20, polysorbate 80 or poloxamer 188.

18. The pharmaceutical composition according to any one of claims 1 to 17, wherein the pharmaceutical composition contains 10 mM histidine, 4% (w / v) sorbitol, 0.02% (w / v) polysorbate 80, 150 mg / ml of the active ingredient, and the pH is adjusted to 5.5 to 6.

5.

19. The pharmaceutical composition according to claim 18, which is for subcutaneous administration.

20. The pharmaceutical composition according to any one of claims 1 to 17, wherein the pharmaceutical composition contains 10 mM histidine, 3.6% (w / v) sorbitol, 0.02% (w / v) polysorbate 80, 10 mg / ml of the active ingredient, and the pH is adjusted to 5.5 to 6.

5.

21. The pharmaceutical composition according to claim 20, which is for intravenous administration.

22. The pharmaceutical composition according to any one of claims 1 to 21, wherein, The active ingredient is A10-1C04. The lyophilized preparation of the pharmaceutical composition according to any one of claims 1 to 22.

24. A pharmaceutical composition comprising a human anti-IL-33 monoclonal antibody as an active ingredient, wherein the combination of the amino acid sequences of the complementarity-determining region 1 (H1), complementarity-determining region 2 (H2), complementarity-determining region 3 (H3) of the heavy chain, complementarity-determining region 1 (L1), complementarity-determining region 2 (L2), and complementarity-determining region 3 (L3) of the light chain of the human anti-IL-33 monoclonal antibody is any one of C1 to C5 in Table 1, the pharmaceutical composition contains a buffer, a nonionic surfactant, and a polyol, [Table 1] 。

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