Pharmaceutical composition containing human Anti-il-33 monoclonal antibody

Abstract

To provide a pharmaceutical composition containing a human anti-IL-33 monoclonal antibody suitable for administration to a subject.SOLUTION: The present invention provides a pharmaceutical composition containing a human anti-IL-33 monoclonal antibody as an active ingredient, the pharmaceutical composition containing essentially no sodium chloride or less than 30 mM sodium chloride. The present invention also provides a freeze-drying agent of the pharmaceutical composition.SELECTED DRAWING: Figure 2

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 an antibody-containing pharmaceutical composition that suppresses turbidity and has improved storage stability, such as pH. [Background technology]

[0002] In recent years, various antibody-containing pharmaceutical compositions have been developed and put into practical use, and most of these compositions are used as intravenous pharmaceutical compositions. However, due to needs in the medical field, there is an increasing demand for the development of antibody-containing pharmaceutical compositions as self-injectable subcutaneous pharmaceutical compositions.

[0003] When designing an antibody-containing pharmaceutical composition for subcutaneous injection, the antibody dose per administration is large (approximately 100 to 200 mg), but the volume of the injection solution is generally limited for subcutaneous injections, making it necessary to increase the antibody concentration in the administration solution. Therefore, highly concentrated antibody-containing pharmaceutical compositions are often prepared using the so-called lyophilization concentration technique, in which a lyophilized pharmaceutical composition is reconstituted using a smaller volume of water than before lyophilization.

[0004] IgG antibodies, a subtype of antibody commonly used for pharmaceutical purposes, are high-molecular-weight glycoproteins of approximately 150 kDa. Because the structure of the antibody variable region (amino acid sequence and sugar chain structure) is highly diverse, each antibody molecular species exhibits different physical properties.

[0005] Antibodies can become denatured through physical stimuli such as heat and vibration, interactions with surfactants, redox agents, sugars, and other compounds, or through long-term storage, causing aggregation, decomposition, and chemical reactions. Denaturation of antibodies can change their binding to antigens and Fc receptors, reducing the antibody's function and effectiveness, and there is concern that aggregated antibodies may cause inflammation. The susceptibility to denaturation, such as aggregation, varies depending on the molecular species of the antibody.

[0006] Because antibodies contain charged amino acids, highly polar amino acids, and sugar chains, they can interact with ions in the antibody solution, causing an imbalanced distribution of ions and resulting in a change in the pH of the antibody solution (pH drift) (the Donnan effect). Changes in the pH of an antibody solution can affect the storage stability of antibodies, but the strength of the Donnan effect varies depending on the molecular species of the antibody.

[0007] Highly concentrated antibody solutions tend to form highly viscous solutions due to the macromolecular nature of proteins and intermolecular interactions. As the viscosity of an antibody solution increases, it becomes difficult to administer the antibody. The increase in viscosity of an antibody solution due to increased antibody concentration varies depending on the molecular species of the antibody.

[0008] Furthermore, when an antibody solution is stored for a long period of time, degradation phenomena such as changes in pH and the formation of insoluble and / or soluble aggregates can become an issue, but these also vary depending on the molecular species of the antibody.

[0009] In general, various efforts have been made to stabilize antibody-containing pharmaceutical compositions so that loss of the active ingredient is minimal even after long-term storage, and pharmaceutical compositions are produced by dissolving the antibody, which is the active ingredient, and various additives such as buffers. However, technologies for preventing antibody aggregation, cloudiness, increased viscosity, and pH drift, particularly in pharmaceutical compositions containing high concentrations of antibodies, are still insufficient.

[0010] We have obtained several human anti-IL-33 monoclonal antibodies that bind to IL-33 (Patent Document 1: WO 2015 / 099175), but there is a need to develop pharmaceutical compositions containing these antibodies that are suitable for administration. [Prior art documents] [Patent documents]

[0011] [Patent Document 1] International Publication No. 2015 / 099175 Summary of the Invention [Problem to be solved by the invention]

[0012] 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. [Means for solving the problem]

[0013] The present inventors have discovered that formulations of human anti-IL-33 monoclonal antibodies become cloudy, and have found that this cloudiness is caused by the formation of aggregates. They have also found that the cloudiness depends on the NaCl concentration and pH. The present invention was completed based on these findings.

[0014] That is, the present invention provides the following. [1] A pharmaceutical composition comprising a human anti-IL-33 monoclonal antibody as an active ingredient, the human anti-IL-33 monoclonal antibody has a heavy chain complementarity-determining region 1 (H1), a heavy chain complementarity-determining region 2 (H2), a heavy chain complementarity-determining region 3 (H3), a light chain complementarity-determining region 1 (L1), a light chain complementarity-determining region 2 (L2), and a light chain complementarity-determining region 3 (L3) in a combination of amino acid sequences selected from any one of C1 to C5 in Table 1; A pharmaceutical composition, wherein said pharmaceutical composition is essentially free of sodium chloride or contains less than 30 mM sodium chloride. [Table 1] [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. [Table 2] [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. [4] The pharmaceutical composition according to any one of [1] to [3], wherein the sodium chloride concentration is 10 mM or less. [5] The pharmaceutical composition according to any one of [1] to [4], which is essentially free of sodium chloride. [6] The pharmaceutical composition according to any one of [1] to [5], whose pH is adjusted to greater than 4 and less than 8. [7] The pharmaceutical composition according to any one of [1] to [6], which has a pH adjusted to 5 or more and 7 or less. [8] The pharmaceutical composition according to any one of [1] to [7], wherein the pH is adjusted with a buffering agent selected from acetate, histidine, and phosphate. [9] The pharmaceutical composition according to any one of [1] to [8], wherein the pH is adjusted with histidine.

[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.

[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.

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

[11] , which contains at least one polyol. [12-1] The pharmaceutical composition described in

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

[13] The pharmaceutical composition according to

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

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

[13] , which contains a surfactant.

[15] The pharmaceutical composition according to

[14] , wherein the surfactant is a nonionic surfactant. [15-1] The pharmaceutical composition described in

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

[16] A pharmaceutical composition according to any one of [1] to [15-1], which contains 10 mM histidine, 4% (w / v) sorbitol, 0.02% (w / v) polysorbate 80, and 150 mg / ml of an active ingredient, and has a pH adjusted to 5.5 to 6.5.

[17] The pharmaceutical composition according to

[16] , for subcutaneous administration.

[18] A pharmaceutical composition according to any one of [1] to [15-1], which contains 10 mM histidine, 3.6% (w / v) sorbitol, 0.02% (w / v) polysorbate 80, and 10 mg / ml of an active ingredient, and has a pH adjusted to 5.5 to 6.5.

[19] The pharmaceutical composition according to

[18] , for intravenous administration.

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

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

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

[20] .

[0015]

[22] A method for treating or preventing an IL-33-associated disease, comprising: administering a pharmaceutical composition containing a human anti-IL-33 monoclonal antibody as an active ingredient to a subject in need thereof; Including, wherein the combination of amino acid sequences of heavy chain complementarity determining region 1 (H1), heavy chain complementarity determining region 2 (H2), heavy chain complementarity determining region 3 (H3), light chain complementarity determining region 1 (L1), light chain complementarity determining region 2 (L2), and 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; wherein the pharmaceutical composition is essentially free of sodium chloride or contains less than 30 mM sodium chloride.

[23] The method described in

[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.

[24] The method according to

[22] or

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

[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.

[26] The method according to any one of

[22] to

[25] , wherein the pharmaceutical composition is essentially free of sodium chloride.

[27] The method according to any one of

[22] to

[26] , wherein the pharmaceutical composition has a pH adjusted to greater than 4 and less than 8.

[28] The method according to any one of

[22] to

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

[29] The method according to any one of

[22] to

[28] , wherein the pharmaceutical composition has a pH adjusted with a buffering agent selected from acetate, histidine, and phosphate.

[30] The method according to any one of

[22] to

[29] , wherein the pharmaceutical composition has a pH adjusted with histidine.

[31] The method according to any one of

[22] to

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

[32] The method according to any one of

[22] to

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

[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.

[34] The method according to any one of

[22] to

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

[35] The method according to any one of

[22] to

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

[36] The method according to

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

[37] The method according to any one of

[22] to

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

[38] The method according to

[37] , wherein the administration is subcutaneous administration.

[39] The method according to any one of

[22] to

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

[40] The method according to

[39] , wherein the administration is intravenous administration.

[41] The method according to any one of

[22] to

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

[0016]

[42] A pharmaceutical composition for use in treating or preventing an IL-33-associated disease, comprising: It contains human anti-IL-33 monoclonal antibody as an active ingredient. wherein the combination of amino acid sequences of heavy chain complementarity determining region 1 (H1), heavy chain complementarity determining region 2 (H2), heavy chain complementarity determining region 3 (H3), light chain complementarity determining region 1 (L1), light chain complementarity determining region 2 (L2), and 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; wherein the pharmaceutical composition is essentially free of sodium chloride or contains less than 30 mM sodium chloride.

[0017]

[43] Use of an IL-33-associated disease in the manufacture of a pharmaceutical composition for the treatment or prevention of the disease, comprising: The pharmaceutical composition comprises a human anti-IL-33 monoclonal antibody and is essentially free of sodium chloride or contains less than 30 mM sodium chloride, wherein the combination of amino acid sequences of heavy chain complementarity determining region 1 (H1), heavy chain complementarity determining region 2 (H2), heavy chain complementarity determining region 3 (H3), light chain complementarity determining region 1 (L1), light chain complementarity determining region 2 (L2), and 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.

[44] The pharmaceutical composition described in [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.

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

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

[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.

[47] The pharmaceutical composition according to any one of

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

[48] ​​The pharmaceutical composition according to

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

[49] The pharmaceutical composition according to

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

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

[51] The pharmaceutical composition according to

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

[52] The pharmaceutical composition according to

[50] or

[51] , which is essentially free of sodium chloride or contains less than 30 mM sodium chloride. [Effects of the Invention]

[0018] The pharmaceutical compositions of the present invention containing the human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04, or A26-1F02) (or pharmaceutical compositions used in the present invention) exhibit reduced cloudiness and are therefore safe and effective. Furthermore, the pharmaceutical compositions of the present invention can be stored stably for long periods in a solution state without significant pH fluctuations, making them suitable for storage and use as formulations. [Brief explanation of the drawings]

[0019] [Figure 1] FIG. 1 shows the cloudiness of the P7N formulation of the A10-1C04 ​​antibody. [Figure 2] Figure 2 shows the effect of adding sugars to suppress cloudiness. Cloudiness was reduced in the A10-1C04 ​​antibody formulations containing sorbitol (A5S, H6S, P6S, and P7S). DETAILED DESCRIPTION OF THE INVENTION

[0020] The following explanation of terms will be given to facilitate understanding of the present invention.

[0021] [Pharmaceutical composition] As used herein, a pharmaceutical composition refers to a composition prepared so that it can be administered to animals such as humans. Therefore, a pharmaceutical composition may also refer to a formulated preparation.

[0022] [Essentially] As used herein, the phrase "essentially free of sodium chloride" refers to a case where sodium chloride is not added to the pharmaceutical composition of the present invention, and refers to a case where the pharmaceutical composition of the present invention does not contain sodium chloride to such an extent that it becomes cloudy.

[0023] [Surfactants] As used herein, surfactant refers to a substance that has a water-compatible portion (hydrophilic group) and an oil-compatible portion (lipophilic / hydrophobic group) in its molecule. Any surfactant commonly used in antibody formulations can be used, including polysorbate 20, polysorbate 80, and poloxamer 188.

[0024] [antibody] The term "antibody" as used herein is used in the broadest sense and includes human antibodies, humanized antibodies, antibodies derived from non-human species, monoclonal antibodies, and polyclonal antibodies. Furthermore, the term "antibody" as used herein may 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, or Fab'.

[0025] [Monoclonal antibody] The term "monoclonal antibody," as used herein, refers to an antibody consisting of a substantially homogeneous population of antibodies, i.e., the individual antibodies comprising the population are identical except for minor differences, e.g., glycosylation or amino acid modifications. In contrast to polyclonal antibodies, which typically include different antibodies, each monoclonal antibody binds to a single epitope on an antigen. The modifier "monoclonal" indicates the character of the antibody as obtained from a substantially homogeneous population of antibodies and is not to be construed as requiring preparation of the antibody by any particular method. For example, monoclonal antibodies to be used in accordance with the present invention may be produced by the hybridoma method first described by Kohler et al., Nature, 256:495 (1975), or may be produced by recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567). The "monoclonal antibodies" may be isolated from phage antibody libraries using the techniques described in Clackson et al., Nature, 352:624-628 (1991) and Marks et al., J. Mol. Biol., 222:581-597 (1991), for example.

[0026] [Five human anti-IL-33 monoclonal antibodies] The five types of human anti-IL-33 monoclonal antibodies used in the present invention are human anti-IL-33 monoclonal antibodies of five clones, A10-1C04, A23-1A05, A25-2C02, A25-3H04, and A26-1F02, disclosed in WO 2015 / 099175, and are human anti-IL-33 monoclonal antibodies consisting of heavy chains and light chains with the following amino acid sequences: (a)A10-1C04: Heavy Chain: EVQLLESGGGLVQPGGSLRLSCAASGFTFSDYYMNWVRQAPGKGLEWVSSISRYSSYIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDIGGMDVWGQGTLVTVS SASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPA PELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 1 in the Sequence Listing); and Light chain: QSVLTQPPSASGTPGQRVTISCTGSSSNIGAVYDVHWYQQLPGTAPKLLIYRNNQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCQTYDSSRWVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO: 2 in the Sequence Listing) (b)A23-1A05: Heavy Chain: EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMHWVRQAPGKGLEWVSSISARSRYHYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARLATRHNAFDIWGQGTLV TVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 3 in the Sequence Listing); and Light chain: QSVLTQPPSASGTPGQRVTISCSGSSSNIGNNAVSWYQQLPGTAPKLLIYASNMRVIGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDDSQKALVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO: 4 in the Sequence Listing) (c)A25-2C02: Heavy Chain: EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMHWVRQAPGKGLEWVSSISARSSYIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARLATRNNAFDIWGQGTLV TVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 5 in the Sequence Listing); and Light chain: QSVLTQPPSASGTPGQRVTISCSGSSSNIGRNAVNWYQQLPGTAPKLLIYASNMRVSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCWAWDDSQKVGVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO: 6 in the Sequence Listing) (d)A25-3H04: Heavy Chain: EVQLLESGGGLVQPGGSLRLSCAASGFTFSRYYMHWVRQAPGKGLEWVSSISAQSSHIYYADSVEGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARLATRQNAFDIWGQGTLV TVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 7 in the Sequence Listing); and Light chain: QSVLTQPPSASGTPGQRVTISCSGSSSNIGRNAVNWYQQLPGTAPKLLIYASNMRRSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCSAWDDSQKVVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO: 8 in the Sequence Listing) (e)A26-1F02: Heavy Chain: EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMHWVRQAPGKGLEWVSSISARSSYLYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARLATRHVAFDIWGQGTLV TVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 9 in the Sequence Listing); and Light chain: QSVLTQPPSASGTPGQRVTISCSGSSSNIGNNAVNWYQQLPGTAPKLLIYASNMRRPGVPDRFSGSKSGTSASLAISGLRSEDEADYYCEAWDDSQKAVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO: 10 in the Sequence Listing)

[0027] [Complementarity-determining region (CDR)] As used herein, the term "complementarity-determining region (CDR)" refers to the amino acid residues of an antibody involved in antigen binding. CDRs, also commonly referred to as "hypervariable regions," are amino acid sequences unique to each antibody molecular species and are identified 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)). Three CDRs (L1, L2, and L3) are present in the antibody light chain, and three CDRs (H1, H2, and H3) are present in the antibody heavy chain. The CDRs of the five clones, A10-1C04, A23-1A05, A25-2C02, A25-3H04, and A26-1F02, are as follows: (a)A10-1C04: H1:DYYMN (SEQ ID NO: 11 in the sequence listing) H2: SISRYSSYIYYADSVKG (SEQ ID NO: 12 in the sequence listing) H3:DIGGMDV (SEQ ID NO: 13 in the sequence listing) L1: TGSSSNIGAVYDVH (SEQ ID NO: 14 in the sequence listing) L2: RNNQRPS (SEQ ID NO: 15 in the sequence listing) L3: QTYDSSRWV (SEQ ID NO: 16 in the sequence listing) (b)A23-1A05: H1:NYYMH (SEQ ID NO: 17 in the sequence listing) H2: SISARSRYHYYADSVKG (SEQ ID NO: 18 in the sequence listing) H3: LATRHNAFDI (SEQ ID NO: 19 in the sequence listing) L1: SGSSSNIGNNAVS (SEQ ID NO: 20 in the sequence listing) L2: ASNMRVI (SEQ ID NO: 21 in the sequence listing) L3: GAWDDSQKALV (SEQ ID NO: 22 in the sequence listing) (c)A25-2C02: H1:NYYMH (SEQ ID NO: 17 in the sequence listing) H2:SISARSSYIYYADSVKG (SEQ ID NO: 23 in the sequence listing) H3:LATRNNAFDI (SEQ ID NO: 24 in the sequence listing) L1:SGSSSNIGRNAVN (SEQ ID NO: 25 in the sequence listing) L2: ASNMRVS (SEQ ID NO: 26 in the sequence listing) L3: WAWDDSQKVGV (SEQ ID NO: 27 in the sequence listing) (d)A25-3H04: H1:RYYMH (SEQ ID NO: 28 in the sequence listing) H2:SISAQSSHIYYADSVEG (SEQ ID NO: 29 in the sequence listing) H3: LATRQNAFDI (SEQ ID NO: 30 in the sequence listing) L1:SGSSSNIGRNAVN (SEQ ID NO: 25 in the sequence listing) L2: ASNMRRS (SEQ ID NO: 31 in the sequence listing) L3: SAWDDSQKVVV (SEQ ID NO: 32 in the sequence listing) (e)A26-1F02: H1:NYYMH (SEQ ID NO: 17 in the sequence listing) H2: SISARSSYLYYADSVKG (SEQ ID NO: 33 in the sequence listing) H3: LATRHVAFDI (SEQ ID NO: 34 in the sequence listing) L1:SGSSSNIGNNAVN (SEQ ID NO: 35 in the sequence listing) L2: ASNMRRP (SEQ ID NO: 36 in the sequence listing) L3: EAWDDSQKAVV (SEQ ID NO: 37 in the sequence listing)

[0028] [Variable area] As used herein, the term "variable region" refers to the portion of a monoclonal antibody other than the constant region, which is involved in antigen binding and determines the antibody specificity, which varies depending on the antigen. The variable region includes heavy chain and light chain variable regions, and the heavy chain and light chain variable regions of the five clones, A10-1C04, A23-1A05, A25-2C02, A25-3H04, and A26-1F02, are as follows: (a) A10-1C04: Heavy chain variable region: EVQLLESGGGLVQPGGSLRLSCAASGFTFSDYYMNWVRQAPGKGLEWVSSISRYSSYIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDIGGMDVWGQGTLVTVSS (SEQ ID NO: 38 in the Sequence Listing) Light chain variable region: QSVLTQPPSASGTPGQRVTISCTGSSSNIGAVYDVHWYQQLPGTAPKLLIYRNNQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCQTYDSSRWVFGGGTKLTVLG (SEQ ID NO: 39 in the Sequence Listing) (b)A23-1A05: Heavy chain variable region: EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMHWVRQAPGKGLEWVSSISARSRYHYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARLATRHNAFDIWGQGTLVTVSS (SEQ ID NO: 40 in the Sequence Listing) Light chain variable region: QSVLTQPPSASGTPGQRVTISCSGSSSNIGNNAVSWYQQLPGTAPKLLIYASNMRVIGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDDSQKALVFGGGTKLTVLG (SEQ ID NO: 41 in the Sequence Listing) (c)A25-2C02: Heavy chain variable region: EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMHWVRQAPGKGLEWVSSISARSSYIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARLATRNNAFDIWGQGTLVTVSS (SEQ ID NO: 42 in the Sequence Listing) Light chain variable region: QSVLTQPPSASGTPGQRVTISCSGSSSNIGRNAVNWYQQLPGTAPKLLIYASNMRVSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCWAWDDSQKVGVFGGGTKLTVLG (SEQ ID NO: 43 in the Sequence Listing) (d)A25-3H04: Heavy chain variable region: EVQLLESGGGLVQPGGSLRLSCAASGFTFSRYYMHWVRQAPGKGLEWVSSISAQSSHIYYADSVEGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARLATRQNAFDIWGQGTLVTVSS (Sequence Number 44 in the Sequence Listing) Light chain variable region: QSVLTQPPSASGTPGQRVTISCSGSSSNIGRNAVNWYQQLPGTAPKLLIYASNMRRSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCSAWDDSQKVVVFGGGTKLTVLG (SEQ ID NO: 45 in the Sequence Listing) (e)A26-1F02: Heavy chain variable region: EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMHWVRQAPGKGLEWVSSISARSSYLYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARLATRHVAFDIWGQGTLVTVSS (SEQ ID NO: 46 in the Sequence Listing) Light chain variable region: QSVLTQPPSASGTPGQRVTISCSGSSSNIGNNAVNWYQQLPGTAPKLLIYASNMRRPGVPDRFSGSKSGTSASLAISGLRSEDEADYYCEAWDDSQKAVVFGGGTKLTVLG (SEQ ID NO: 47 in the Sequence Listing)

[0029] "Stability" As used herein, "stable" means that an antibody-containing pharmaceutical composition substantially retains its properties (e.g., physical properties, chemical properties, and / or biological activity) after storage. For example, various analytical techniques for measuring the stability of proteins, including antibodies, are available in the art and are reviewed 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 an antibody-containing pharmaceutical composition can be evaluated at a selected temperature for a selected period of time. A "stable" antibody-containing pharmaceutical composition is one in which no significant changes are observed at refrigerated temperatures (2-8°C) for at least 1 month, 3 months, 6 months, or 12 months, preferably 2 years, and more preferably 3 years; at room temperature (23-27°C) for at least 3 months, preferably 6 months, and more preferably 1 year; or under stress conditions (approximately 40°C or approximately 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 (e.g., turbidity), pH, viscosity, antibody antigen-binding ability, antibody inhibitory activity against the antigen molecule (e.g., inhibitory activity against IL-6 production induction by IL-33), antibody effector function, and antibody degradation.

[0030] Effector function Antibody "effector functions" refer to those biological activities attributable to the Fc region of an antibody (either a native sequence Fc region or an amino acid sequence variant Fc region). Examples of antibody effector functions include C1q binding, complement dependent cytotoxicity, Fc receptor binding, antibody-dependent cell-mediated cytotoxicity (ADCC), phagocytosis, down-regulation of cell surface receptors (e.g., B cell receptor, BCR), etc.

[0031] [Cloudy] As used herein, "cloudiness" refers to a cloudy white state as determined by visual inspection of color and / or transparency (turbidity). Cloudiness can be further analyzed by measuring fine particles using a flow cytometer particle image analyzer or particle counter, measuring interaction parameters (hereinafter simply referred to as "interaction parameters" or "Kd values") using dynamic light scattering (DLS), or measuring turbidity (absorbance at 650 nm (OD650)). Analytical values ​​correlated with "cloudiness" include, for example, an OD650 of 0.009, 0.010, 0.011, 0.012, 0.013, or 0.014 or greater for the antibody-containing pharmaceutical composition; a Kd value of 0, -1, -2, -3, or -4 mL / g or less; and / or a particle count of 500, 750, 1000, 1250, or 1500 particles / mL or greater measuring 1.5 μm or greater as determined by a particle counter.

[0032] [Lyophilized formulation] As used herein, the term "lyophilized formulation" refers to a dry (e.g., freeze-dried) pharmaceutical composition that is substantially free of water. Techniques for lyophilizing antibodies are well known in the art; see, for example, Rey & May (2004) Freeze-Drying / Lyophilization of Pharmaceutical & Biological Products ISBN 0824748689.

[0033] [IL-33] IL-33 is a cytokine belonging to the IL-1 family and is also known as NF-HEV. When released extracellularly, IL-33 binds to IL-33 receptors (ST2 and IL-1RAcP) and initiates intracellular signal transduction in cells expressing the IL-33 receptor. IL-33-induced signal transduction includes, but is not limited to, the NF-κB pathway and the MAPKKs pathway, ultimately inducing 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, and IL-13, with IL-5, IL-6, and IL-13 being particularly induced. Examples of chemokines induced by IL-33 include CXCL2, CCL2, CCL3, CCL6, CCL17, and CCL24. Examples of inflammatory mediators induced by IL-33 include PGD2 and LTB4. Cytokines, chemokines, and inflammatory mediators induced by IL-33 are involved in immune cell migration, cytokine production, and degranulation, causing inflammation. In the present invention, the term "IL-33" refers to either full-length IL-33 or mature IL-33, or to homologous derivatives or variants thereof, as long as the "five types of human anti-IL-33 monoclonal antibodies" bind to and inhibit at least one of the above functions. Furthermore, the term "IL-33" may refer to either full-length IL-33 or mature IL-33, or to homologous derivatives or variants thereof. Furthermore, the term "IL-33" may refer to either human IL-33 or IL-33 derived from other organisms.

[0034] [Pharmaceutically acceptable] As used herein, the term "pharmaceutically acceptable" means does not interfere with the effectiveness of the biological activity of the active ingredient(s) and is non-toxic.

[0035] [Isotonic] As used herein, an "isotonic" formulation has an osmotic pressure substantially identical to that of human blood. Isotonic pharmaceutical compositions generally have an osmotic pressure ratio relative to blood of about 0.9 to 1.2. Osmotic pressure can be measured, for example, using a vapor pressure or ice-freezing osmometer.

[0036] [pH drift] As used herein, the term "pH drift" refers to a change in the pH value of the pharmaceutical composition of the present invention before and after treatment such as storage or concentration.

[0037] Hereinafter, embodiments of the present invention will be described. Note that the following embodiments are merely examples for explaining the present invention, and the present invention is not limited to these embodiments.

[0038] The human anti-IL-33 monoclonal antibody of the present invention can be produced by known techniques, for example, the method described in Patent Document 1.

[0039] The monoclonal antibodies produced as described above can be formulated into desired compositions by methods such as dialysis, ultrafiltration, ammonium sulfate precipitation, etc. Alternatively, the monoclonal antibodies can be prepared into a solution containing a desired buffer, and then sugars, surfactants, etc. can be added thereto to form a formulation.

[0040] The present inventors have found that when five types of human anti-IL-33 monoclonal antibodies are prepared in solution, they partially aggregate, causing the solution to become cloudy. This cloudiness is due to the formation of antibody-containing aggregate microparticles. When these antibodies are formulated into pharmaceutical compositions, there are concerns that they may reduce the biological activity of the antibody (active ingredient), worsen pharmacokinetics due to the induction of anti-drug antibodies (ADA) by highly immunogenic aggregates, and further cause inflammation due to the aggregates themselves. Therefore, it is necessary to suppress this cloudiness. The present invention relates to pharmaceutical compositions in which cloudiness is suppressed. The pharmaceutical compositions of the present invention contain five types of human anti-IL-33 monoclonal antibodies as active ingredients, and may further contain salts, buffers, surfactants, sugars, and the like.

[0041] [Salt concentration] The present inventors have found that the cloudiness of the five human anti-IL-33 monoclonal antibodies is primarily due to salt. Therefore, the pharmaceutical composition of 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, or 1 mM or less, and more preferably essentially salt-free. The pharmaceutical composition of the present invention also preferably has a low salt concentration, preferably less than 50 mM, 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, or 1 mM, and more preferably essentially salt-free. Examples of salts present at low concentrations or essentially free of salt in the pharmaceutical composition of the present invention include inorganic salts and organic salts. Examples of inorganic salts include, inter alia, sodium chloride, potassium chloride, magnesium chloride, calcium chloride, sodium sulfate, potassium sulfate, magnesium sulfate, calcium sulfate, with sodium chloride being preferred.

[0042] [Buffers and pH] The pH of the pharmaceutical composition of the present invention is adjusted using a buffer. Suitable buffers for pharmaceutical compositions include, but are not limited to, gluconate, histidine, citrate, phosphate (e.g., sodium or potassium), succinate (e.g., sodium), acetate, trishydroxymethylaminomethane, glycine, arginine, and combinations thereof. Different buffers can be used depending on the pH to be adjusted. The pharmaceutical composition of the present invention preferably contains acetate, histidine, or phosphate as a buffer, and more preferably histidine. The concentration of the buffer is not particularly limited as long as it is pharmaceutically acceptable, but is preferably 1 mM to 150 mM, more preferably 5 mM to 100 mM, and even more preferably 10 mM to 50 mM. The buffer concentration 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. When using a buffer containing a salt, particularly an inorganic salt, it is preferable to use a concentration of less than 30 mM, particularly 10 mM or less.

[0043] Histidine (e.g., at a concentration of 5 mM to 50 mM, e.g., 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, or about 50 mM) is particularly useful as a buffering agent for the pharmaceutical composition of the present invention. In one embodiment, the stable pharmaceutical composition contains 5 mM to 20 mM histidine. The pH of the pharmaceutical composition may be within the range of 4.0 to 8.0, and is generally within the range of 4.5 to 7.5, for example, 5.0 to 7.0, 5.2 to 6.8, such as 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. The pharmaceutical composition of the present invention increases in turbidity upon long-term storage at pH 4 or 8. Furthermore, the pharmaceutical composition of the present invention undergoes significant pH drift upon long-term storage at pH 4 or 8. Thus, in one embodiment, the pH of a stable antibody-containing pharmaceutical composition is greater than 4 and less than 8, preferably 5 or greater than 7 or less, more preferably 5.5 or greater than 6.5 or less, and most preferably 6.0.

[0044] [Surfactants] The pharmaceutical composition of the present invention preferably comprises a surfactant. Suitable surfactants for pharmaceutical compositions include, but are not limited to, nonionic surfactants, ionic surfactants, zwitterionic surfactants, and combinations thereof. Common surfactants for use in the present invention include, but are not limited to, sorbitan fatty acid esters (e.g., sorbitan monocaprylate, sorbitan monolaurate, sorbitan monopalmitate), sorbitan trioleate, glycerin fatty acid esters (e.g., glycerin monocaprylate, glycerin monomyristate, glycerin monostearate), polyglycerin fatty acid esters (e.g., decaglyceryl monostearate, decaglyceryl distearate, decaglyceryl monolinoleate), 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 trioleate), polyoxyethylene sorbitol tristearate, polyoxyethylene sorbitol trioleate, polyoxyethylene sorbitol tristearate), polyoxyethylene sorbitol trioleate, ... tetraoleate), polyoxyethylene glycerin fatty acid esters (e.g., polyoxyethylene glyceryl 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 nonylphenyl 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 stearic acid amide), C10-C18 alkyl sulfates (e.g., sodium cetyl sulfate, sodium lauryl sulfate, sodium oleyl sulfate), polyoxyethylene C10-C18 alkyl ether sulfates having an average of 2-4 moles of ethylene oxide added (e.g., polyoxyethylene sodium lauryl sulfate), and C1-C18 alkyl sulfosuccinate salts (e.g., sodium lauryl sulfosuccinate), as well as natural surfactants such as lecithin, glycerophospholipids, sphingophospholipids (e.g., sphingomyelin), and sucrose esters of C12-C18 fatty acids. The pharmaceutical composition of the present invention can contain one or more of these surfactants.Preferred surfactants are nonionic surfactants (e.g., sorbitan fatty acid esters, sorbitan trioleate, glycerin fatty acid esters, polyglycerin fatty acid esters, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene sorbitol fatty acid esters, polyoxyethylene glycerin 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 surfactant concentration may be any concentration commonly used in the art, for example, about 0.01% (w / v) to about 0.1% (w / v), such as 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 surfactants, polysorbate 80 (Tween 80) is particularly useful. 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.

[0045] [Polyol] The pharmaceutical composition of the present invention preferably contains a polyol, and the addition of a polyol can adjust the osmotic pressure of the pharmaceutical composition and also suppress the formation of aggregates. The polyol contained in the pharmaceutical composition of the present invention is not particularly limited as long as it is pharmaceutically acceptable, but a polyol that dissolves at least 100 g in 100 g of water at 20°C (i.e., a polyol with a solubility in water at 20°C of 100 g / 100 g or more) is preferred. Polyols are also called polyhydric alcohols, and may be any molecule having two or more alcoholic hydroxyl groups. Examples of polyols include glycerol (glycerin), propylene glycol, polyethylene glycol (PEG), and sugars, with sugars being preferred. Sugars suitable for use in the pharmaceutical compositions of the present invention include, but are not limited to, compounds having the general formula (CHO) and derivatives thereof, including monosaccharides, disaccharides, trisaccharides, polysaccharides, sugar alcohols, reducing sugars, non-reducing sugars, etc. 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. The sugar contained in the pharmaceutical composition of the present invention is preferably sorbitol, sucrose, trehalose, or mannitol, and most preferably sorbitol or sucrose. The concentration of sugars contained in the pharmaceutical composition of the present invention is not particularly limited as long as it is pharmaceutically acceptable, but is preferably 50 mM to 300 mM, more preferably 165 mM to 275 mM, and even more preferably 200 mM to 220 mM. Furthermore, the sugar concentration 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, or about 300 mM. In one embodiment, the stable pharmaceutical composition contains 3% to 5% (w / v) (165 to 275 mM) sorbitol. In another embodiment, the stable pharmaceutical composition contains about 3.6% (w / v) or about 4% (w / v) sorbitol (about 200 mM or about 220 mM, respectively).

[0046] The osmotic pressure ratio of the pharmaceutical composition of the present invention is preferably 0.5 to 4, more preferably 0.7 to 3, even more 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, or 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, it is preferable to adjust the osmotic pressure of the pharmaceutical composition using a polyol.

[0047] [Human anti-IL-33 monoclonal antibody] The pharmaceutical compositions of the present invention contain 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, or 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, or about 175 mg / mL. The antibody concentration of the pharmaceutical composition of the present invention is preferably less than 175 mg / mL, and more preferably 150 mg / mL or less, due to high viscosity at 175 mg / mL or higher. In one embodiment, a stable pharmaceutical composition contains about 10 mg / mL or about 150 mg / mL of A10-1C04. In one embodiment, a stable pharmaceutical composition contains about 10 mg / mL or about 150 mg / mL of A23-1A05. In one embodiment, a stable pharmaceutical composition contains about 10 mg / mL or about 150 mg / mL of A25-2C02. In one embodiment, a stable pharmaceutical composition contains about 10 mg / mL or about 150 mg / mL of A25-3H04. In one embodiment, a stable pharmaceutical composition contains about 10 mg / mL or about 150 mg / mL of A26-1F02.

[0048] The pharmaceutical compositions of the present invention preferably have an appropriate viscosity so that they can be easily administered to any patient. A low viscosity allows for administration without the need for strong force, and the inventors have found that a viscosity of about 20 cP or higher for the pharmaceutical compositions of the present invention makes them somewhat difficult to eject from the syringe during administration. Therefore, the viscosity of the pharmaceutical compositions of the present invention is preferably 50 cP or less, more preferably 30 cP or less, even more preferably 20 cP or less, and most preferably 10 cP or less. In one embodiment, the viscosity of the pharmaceutical compositions of the present invention is about 1, about 2, about 5, about 10, about 15, or about 20 cP. The viscosity of the pharmaceutical compositions of the present invention can be measured by rheometry (rotational or capillary).

[0049] [Method of using the pharmaceutical composition] The pharmaceutical composition of the present invention is used for treating (treating, preventing, etc.) patients with IL-33-related diseases such as asthma, allergies (atopic dermatitis, hay fever), and endometriosis. The mode of administration is not particularly limited, and may be systemic or local. Examples include intravenous, subcutaneous, intramuscular, and intraperitoneal administration. For example, if the pharmaceutical composition of the present invention is administered subcutaneously, the dosage is limited, so it preferably contains a high concentration of human anti-IL-33 antibody. For example, a pharmaceutical composition containing 10 mM histidine, 4% (w / v) sorbitol, 0.02% (w / v) polysorbate 80, and 150 mg / ml of the active ingredient, with a pH adjusted to 5.5 to 6.5, is preferred. For example, when administered intravenously, the pharmaceutical composition of the present invention may contain a low concentration of human anti-IL-33 antibody, and is preferably a pharmaceutical composition containing, for example, 10 mM histidine, 3.6% (w / v) sorbitol, 0.02% (w / v) polysorbate 80, and 10 mg / ml of the active ingredient, with the pH adjusted to 5.5 to 6.5.

[0050] Examples of IL-33-associated diseases include, but are not limited to, asthma, atopic dermatitis, urticaria, hay fever, anaphylactic shock, eosinophilic sinusitis, hypereosinophilic syndrome, 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, and 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, Sjogren's syndrome, Behçet's disease, pemphigus, 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 greata), antiphospholipid syndrome, autoimmune disorders of the adrenal gland (e.g., autoimmune Addison's disease), celiac sprue dermatitis, chronic fatigue and immune dysfunction syndrome (CFIDS), cold agglutinin disease, essential mixed cryoglobulinemia, fibromyalgia-fibromyositis, glomerulonephritis (e.g., IgA 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, polychondritis rondritis), polyglandular syndrome, stiff-man syndrome, vitiligo, sarcoidosis, polyendocrinopathy, other endocrine gland deficiencies, 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 disease, post-myocardial infarction (post-MI), uveitis, uveitis ophthalmiaOpthalmia), vasculitis, primary agammaglobulinemia, cancer (e.g., brain tumor, laryngeal cancer, lip and oral cavity cancer, hypopharyngeal cancer, thyroid cancer, esophageal cancer, breast cancer, lung cancer, stomach cancer, adrenocortical carcinoma, bile duct cancer, 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's tumor, Hodgkin's disease, non-Hodgkin's lymphoma, melanoma, mesothelioma, multiple myeloma, etc.), immune system Examples of such diseases include infections that resist elimination by the immune system (e.g., severe acute respiratory syndrome (SARS)), fatal cytokine storms associated with highly virulent influenza infections, and sepsis. Preferred examples include asthma, atopic dermatitis, hay fever, anaphylactic shock, scleroderma, Crohn's disease, ulcerative colitis, arthritis, systemic lupus erythematosus, ankylosing spondylitis, hepatic fibrosis, pulmonary fibrosis, acute kidney injury, vasculitis, and cancer.

[0051] A "stable" pharmaceutical composition of the present invention exhibits no significant change for at least 12 months, preferably 2 years, and more preferably 3 years, at refrigerated temperatures (2-8°C), or for at least 3 months, preferably 6 months, and more preferably 1 year, at room temperature (22-28°C). For example, after 2 years of storage at 5°C, no turbidity is observed, the OD650 is 0.014 or less, preferably 0.01 or less, and more preferably 0.008 or less, the pH drift is 1 or less, preferably 0.8 or less, and more preferably 0.5 or less, the Kd value is -4 mL / g or more, preferably -2 mL / g or more, and more preferably a positive value, and the number of particles of 1.5 μm or larger measured by a particle counter is 1500 particles / mL or less, preferably 1000 particles / mL or less, more preferably 750 particles / mL or less, and most preferably 500 particles / mL or less.

[0052] If necessary, preservatives, anti-adsorption agents, soothing agents, sulfur-containing reducing agents, antioxidants, etc. may be added appropriately to the pharmaceutical composition of the present invention.

[0053] 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.

[0054] The adsorption inhibitor 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.

[0055] The soothing agent is not particularly limited as long as it is pharmaceutically acceptable, and examples thereof include local anesthetics such as lidocaine.

[0056] The sulfur-containing reducing agent is not particularly limited as long as it is pharmaceutically acceptable, and examples thereof include those having a sulfhydryl group, such as N-acetylcysteine, N-acetylhomocysteine, thioctic acid, thiodiglycol, thioethanolamine, thioglycerol, thiosorbitol, thioglycolic acid and its salts, sodium thiosulfate, glutathione, and thioalkanoic acids having 1 to 7 carbon atoms.

[0057] The antioxidant is not particularly limited as long as it is pharmaceutically acceptable, and examples thereof include erythorbic acid, dibutylhydroxytoluene, butylhydroxyanisole, α-tocopherol, tocopherol acetate, L-ascorbic acid and its salts, L-ascorbyl palmitate, L-ascorbyl stearate, sodium bisulfite, sodium sulfite, triamyl gallate, propyl gallate, and chelating agents such as disodium ethylenediaminetetraacetate (EDTA), sodium pyrophosphate, and sodium metaphosphate.

[0058] The pharmaceutical composition of the present invention may be a freeze-dried formulation for longer storage.

[0059] 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. [Example]

[0060] Comparative Example 1: Cloudiness of pharmaceutical composition containing human anti-IL-33 monoclonal antibody Human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04, or A26-1F02) were prepared in a solvent (10 mM Na-phosphate / pH 7 / 150 mM NaCl / 0.02% (w / v) Polysobate 80) (hereinafter abbreviated as "P7N") at a concentration of 150 mg / mL. The composition was visually inspected for cloudiness (e.g., Figure 1 shows A10-1C04). Cloudiness was confirmed by visual observation under white light (13 W fluorescent lamp) against a black background. Subvisible particles (fine particles 5 μm or larger) were measured for this pharmaceutical composition using a FlowCam (Fluid Imaging Technologies), revealing 1919 particles / mL. These results suggest that the human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04, or A26-1F02) have high aggregation properties and are difficult to formulate.

[0061] Comparative Example 2: Effect of pH on turbidity of pharmaceutical compositions containing human anti-IL-33 monoclonal antibodies To eliminate the cloudiness of the human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04, or A26-1F02), the pH of the pharmaceutical composition was altered by changing the buffer as described below based on the formulation of Comparative Example 1. The antibody concentration was 150 mg / ml. The formulations investigated are shown below. 10 mM Na-acetate / pH 4 / 150 mM NaCl / 0.02% (w / v) Polysobate 80 (hereafter referred to as "A4N") 10 mM Na-acetate / pH 5 / 150 mM NaCl / 0.02% (w / v) Polysobate 80 (hereafter referred to as "A5N") 10 mM Histidine / pH 6 / 150 mM NaCl / 0.02% (w / v) Polysobate 80 (hereafter referred to as "H6N") 10 mM Na-phosphate / pH 6 / 150 mM NaCl / 0.02% (w / v) Polysobate 80 (hereafter referred to as "P6N") 10 mM Na-phosphate / pH 7 / 150 mM NaCl / 0.02% (w / v) Polysobate 80 (hereinafter referred to as "P7N") and 10 mM Na-phosphate / pH 8 / 150 mM NaCl / 0.02% (w / v) Polysobate 80 (hereafter referred to as "P8N") Evaluation of the properties showed that the formulation became cloudy at pH levels of 4, 5, 6, 7, and 8. This result suggests that changing the pH of the formulation would not improve the cloudiness. The properties were confirmed with the naked eye by the method described in Comparative Example 1.

[0062] Example 1: Effect of adding sugar on cloudiness suppression To eliminate the cloudiness of human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04, or A26-1F02), evaluation was performed using the following formulations in which 5% sorbitol was added instead of the sodium chloride contained in the formulations A4N, A5N, H6N, P6N, P7N, and P8N of Comparative Example 2. The antibody concentration was 150 mg / ml. 10 mM Na-acetate / pH 4 / 5% (w / v) Sorbitol / 0.02% (w / v) Polysobate 80 (hereafter referred to as "A4S") 10 mM Na-acetate / pH 5 / 5% (w / v) Sorbitol / 0.02% (w / v) Polysobate 80 (hereafter referred to as "A5S") 10mM Histidine / pH6 / 5%(w / v) Sorbitol / 0.02%(w / v) Polysobate 80 (hereafter referred to as "H6S") 10mM Na-phosphate / pH 6 / 5% (w / v) sorbitol / 0.02% (w / v) polysobate 80 (hereafter referred to as "P6S") 10 mM Na-phosphate / pH 7 / 5% (w / v) sorbitol / 0.02% (w / v) Polysobate 80 (hereinafter referred to as "P7S") 10 mM Na-phosphate / pH 8 / 5% (w / v) sorbitol / 0.02% (w / v) Polysobate 80 (hereinafter referred to as "P8S") Evaluation of the properties and OD650 (turbidity) showed that adding sorbitol instead of sodium chloride improved the turbidity (for example, Figure 2 shows the results for A10-1C04). Furthermore, the addition of sorbitol inhibited 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). The properties were confirmed with the naked eye by the method described in Comparative Example 1. OD650 was measured by measuring the absorbance at 650 nm for 100 μL of each formulation solution using a Molecular Device microplate reader (SoftMax Pro software).

[0063] [Table 3]

[0064] Example 2: Salt concentration and turbidity (1) Human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04, or A26-1F02) were dissolved in 10 mM histidine buffer (pH 6.0 or pH 5.5) and added with 0, 50, or 100 mM NaCl to measure the interaction parameter (Kd value), an index of aggregation. The measurement temperature was 25°C, and antibody concentrations were adjusted to 0.5, 1, 2.5, 5, 10, or 20 mg / mL. As a result, Kd values ​​became negative (increased aggregation) at NaCl concentrations of 50 mM or more, suggesting that NaCl concentrations of less than 50 mM are preferable (e.g., Table 4 for A10-1C04). The Kd value can be calculated by determining the diffusion coefficient (Dm) using dynamic light scattering, finding the slope from a plot of antibody concentration (horizontal axis) and diffusion coefficient (vertical axis), and dividing this slope by the diffusion coefficient (D0) at zero concentration. That is, the Kd value is calculated using the following relational formula: The larger the positive Kd value, the lower the agglutination tendency. Dm = D0 (1 + Kd value × [antibody concentration])

[0065] [Table 4]

[0066] Example 3: Salt concentration and turbidity (2) The Kd value, an index of agglutination, was measured by adding 0, 5, 10, or 30 mM NaCl to 10 mM histidine / pH 6.0 / 3.6% (w / v) sorbitol. The antibody A10-1C04 ​​was used at 2.5, 5, 10, or 14 mg / mL. The Kd value became negative at NaCl concentrations of 30 mM or higher (Kd values ​​of 17.3, 7.3, 1.2, and -4.0 mL / g for NaCl concentrations of 0, 5, 10, and 30 mM, respectively). Therefore, it was suggested that NaCl concentrations of less than 30 mM are preferable. The Kd value was calculated using the method described in Example 2.

[0067] Example 4: pH Drift The pH transition during long-term storage of pharmaceutical compositions containing human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04, or A26-1F02) was measured. The antibody concentration was 150 mg / mL, and formulations with pH values ​​ranging from 4 to 8 were examined (A4S, A5S, H6S, P6S, P7S, and P8S). The pH of each formulation was measured at the time of preparation and after storage at 40°C for 2, 4, 8, and 12 weeks. Significant pH drift was observed in formulations with pH values ​​of 4 (A4S) and 8 (P8S) (see Table 5 for A10-1C04, for example). Therefore, formulations with pH values ​​of 5 to 7, which have minimal pH drift, were considered preferable.

[0068] [Table 5]

[0069] Example 5: Antibody concentration and viscosity Pharmaceutical compositions were prepared containing human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04, or A26-1F02) at antibody concentrations of 15, 50, 100, 125, 150, 175, or 200 mg / ml in a 10 mM histidine / pH 6 / 3.6% (w / v) sorbitol / 0.02% (w / v) Polysobate 80 formulation. Viscosity at 25°C was measured using a Brookfield DV3TLVCJ0 viscometer (CPA-40Z spindle, 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 viscosity increases rapidly above 150 mg / mL. Furthermore, when solutions of each viscosity were prepared and their ejection properties were examined using a syringe, it was found that administration became somewhat difficult when the viscosity exceeded approximately 20 cP, suggesting that an antibody concentration of less than 175 mg / mL is appropriate.

[0070] Example 6: Sugar addition and evaluation of aggregation and particle generation suppression effect Compositions containing human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04, or A26-1F02) in 10 mM histidine / pH 6.0 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, an index of aggregation, was examined. Kd values ​​were calculated as described in Example 2. For example, when A10-1C04 ​​was used, the Kd value was 33.4 mL / g without added sugar, and 24.7 mL / g and 33.5 mL / g with the addition of 3.6% (w / v) sucrose or sorbitol, respectively, both of which were positive values. These results suggested that adding sucrose instead of sorbitol would be acceptable. In addition, for a formulation containing 10 mg / ml antibody and 3.6% (w / v) sorbitol (10 mM histidine, pH 6.0, 3.6% (w / v) sorbitol), the number of particles 1.5 μm or larger was measured using a particle counter (HIAC, HACH System 9703+) at the time of preparation and after one week of storage at 50°C. HIAC measurements were performed four times for each sample using an injection volume of 100 μL, and the data from the first run was discarded. The results showed that the addition of 3.6% (w / v) sorbitol suppressed the increase in particle count compared to the absence of sorbitol, suggesting that the addition of sorbitol has an inhibitory effect on aggregation (e.g., Table 6 for A10-1C04).

[0071] [Table 6]

[0072] Example 7: Aggregation suppression effect of surfactants A 10 mg / mL solution 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 aggregation was examined. No aggregation was observed at the time of preparation, regardless of whether a surfactant was added or not.

[0073] Example 8: Stability of subcutaneous and intravenous formulations Human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04, or A26-1F02) were prepared as intravenous formulations (10 mg / mL antibody / 10 mM histidine / pH 6 / 3.6% (w / v) sorbitol / 0.02% (w / v) polysorbate 80) and subcutaneous formulations (150 mg / mL antibody / 10 mM histidine / pH 6 / 4% (w / v) sorbitol / 0.02% (w / v) polysorbate 80). The intravenous formulations were examined and found to be stable, with no cloudiness, increase in aggregated particle count, or pH drift observed after 24 months of storage at 5°C. The stability of the subcutaneous formulations can also be evaluated in a similar manner.

[0074] Example 9: Effect of pH on turbidity of pharmaceutical compositions containing human anti-IL-33 monoclonal antibodies (2) To eliminate the cloudiness of the human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04, or A26-1F02), the following formulation was prepared, which did not contain sodium chloride and was within the pH range of the formulation confirmed in Comparative Example 2. 10mM Na-acetate / pH4 / 0.02% (w / v) Polysorbate 80 (hereafter referred to as "A4") 10mM Na-acetate / pH5 / 0.02% (w / v) Polysorbate 80 (hereafter referred to as "A5") 10mM Histidine / pH6 / 0.02% (w / v) Polysorbate 80 (hereafter referred to as "H6") 10mM Histidine / pH7 / 0.02% (w / v) Polysorbate 80 (hereafter referred to as "H7") 10mM Na-phosphate / pH8 / 0.02% (w / v) Polysorbate 80 (hereinafter referred to as "P8") For example, when the properties of 150 mg / mL A10-1C04 ​​were evaluated, the cloudiness was improved by not adding sodium chloride, and the inhibitory effect was particularly good at pH 4 to 7 (Table 7).

[0075] [Table 7]

[0076] Example 10: Effect of salt on turbidity of pharmaceutical compositions containing human anti-IL-33 monoclonal antibodies (2) To eliminate the cloudiness of the 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 evaluation was performed. 10mM Na-acetate / pH4 / 10mM NaCl / 0.02% (w / v) Polysorbate 80 (hereafter referred to as "A4N10") 10mM Na-acetate / pH4 / 30mM NaCl / 0.02% (w / v) Polysorbate 80 (hereafter referred to as "A4N30") 10mM Na-acetate / pH4 / 50mM NaCl / 0.02% (w / v) Polysorbate 80 (hereafter referred to as "A4N50") 10mM Na-acetate / pH4 / 100mM NaCl / 0.02% (w / v) Polysorbate 80 (hereafter referred to as "A4N100") 10mM Na-acetate / pH 5 / 10mM NaCl / 0.02% (w / v) Polysorbate 80 (hereafter referred to as "A5N10") 10mM Na-acetate / pH5 / 30mM NaCl / 0.02% (w / v) Polysorbate 80 (hereafter referred to as "A5N30") 10mM Na-acetate / pH5 / 50mM NaCl / 0.02% (w / v) Polysorbate 80 (hereafter referred to as "A5N50") 10mM Na-acetate / pH 5 / 100mM NaCl / 0.02% (w / v) Polysorbate 80 (hereafter referred to as "A5N100") 10mM Histidine / pH6 / 10mM NaCl / 0.02% (w / v) Polysorbate 80 (hereafter referred to as "H6N10") 10mM Histidine / pH6 / 30mM NaCl / 0.02% (w / v) Polysorbate 80 (hereafter referred to as "H6N30") 10mM Histidine / pH6 / 50mM NaCl / 0.02% (w / v) Polysorbate 80 (hereafter referred to as "H6N50") 10mM Histidine / pH6 / 100mM NaCl / 0.02% (w / v) Polysorbate 80 (hereafter referred to as "H6N100") For example, when the properties of 150 mg / mL A10-1C04 ​​were evaluated, cloudiness was suppressed at 10 mM sodium chloride at pH 4 to 6, but cloudiness was observed at sodium chloride concentrations above 30 mM (Table 8). [Table 8]

[0077] Example 11: Aggregation suppression effect of surfactants (2) To assess the effect of surfactants on 10 mg / mL human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04, or A26-1F02), the following formulations containing surfactants were prepared. These samples were then rotated at 30 rpm in a 50°C incubator using a rotator. Samples taken over time were visually inspected for 5 seconds against a white or black board under fluorescent light of 2000 to 3750 Lux. Numerous aggregates were observed after 2 days in the surfactant-free system, but no aggregates were observed even after 7 days in the samples containing polysorbate 80 or poloxamer 188. In particular, no aggregates were observed even after 14 days in the systems containing 0.02% or more polysorbate 80. In the sample to which polysorbate 20 was added, no foreign matter was observed up to 4 days later, a small amount of foreign matter was observed after 7 days, but no foreign matter was observed after 14 days. For example, the results of the test on A10-1C04 ​​are shown in Table 9. 10mM Histidine / pH6 (hereafter abbreviated as "H6(-)") 10mM Histidine / pH6 / 0.02%(w / v) Poloxamer 188 (hereafter abbreviated as "H6PX") 10mM Histidine / pH6 / 0.02%(w / v)Polysorbate20 (hereafter referred to as "H6P20") 10mM Histidine / pH6 / 0.01%(w / v)Polysorbate80 (hereafter referred to as "H6PS1") 10mM Histidine / pH6 / 0.02%(w / v)Polysorbate80 (hereafter referred to as "H6PS2") 10mM Histidine / pH6 / 0.05%(w / v)Polysorbate80 (hereafter referred to as "H6PS5") [Table 9]

[0078] Example 12: Sugar addition and aggregation evaluation (2) A 10 mg / mL solution 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 was prepared, and mannitol, trehalose, sucrose, or sorbitol was added to prepare a drug solution. These samples were then frozen at -80°C for at least 8 hours and then left at room temperature for at least 4 hours before thawing. This process was repeated six times. The samples were then visually inspected for 5 seconds each against a white or black board under fluorescent light of 2000 to 3750 lux. The addition of sugar reduced the amount of aggregates after freeze-thawing compared to the non-additive system, and aggregation was particularly suppressed in the systems using sorbitol and sucrose. For example, the results for A10-1C04 ​​are shown in Table 10. 10mM Histidine / pH6 (hereafter abbreviated as "H6(-)") 10mM Histidine / pH6 / 3.0%(w / v) Sorbitol (hereafter referred to as "H6So3") 10mM Histidine / pH6 / 3.6%(w / v) Sorbitol (hereafter referred to as "H6So3.6") 10mM Histidine / pH6 / 4.0%(w / v) Sorbitol (hereafter referred to as "H6So4") 10mM Histidine / pH6 / 5.0%(w / v) Sorbitol (hereafter referred to as "H6So5") 10mM Histidine / pH6 / 3.6%(w / v)Sucrose (hereafter abbreviated as "H6Su") 10mM Histidine / pH6 / 3.6%(w / v)Trehalose (hereafter abbreviated as "H6Tr") 10mM Histidine / pH6 / 3.6%(w / v)Mannitol (hereafter abbreviated as "H6Ma")

[0079] [Table 10]

[0080] Example 13: Freeze-drying A 150 mg / mL solution 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, and 0.02% (w / v) Polysorbate 80 was prepared. Mannitol, trehalose, sucrose, or sorbitol was added, and the mixture was freeze-dried using a tray freeze dryer (Kyowa Vacuum). The shape of the freeze-dried cake was visually inspected. Resolubility was evaluated by adding water for injection, leaving the solution at 5°C for half a day, and then checking whether a cake-like form remained in the solution. All samples had a good cake-like form, and resolubility after adding water for injection was confirmed. It was confirmed that preparation as a lyophilized formulation is possible. For example, the results for A10-1C04 ​​are shown in Table 11. 10mM Histidine / pH6 / 0.02% (w / v) Polysorbate 80 / 4.0% (w / v) Sorbitol (hereafter abbreviated as "LYSO") 10mM Histidine / pH6 / 0.02% (w / v) Polysorbate 80 / 4.0% (w / v) Sucrose (hereafter abbreviated as "LYSU") 10mM Histidine / pH6 / 0.02% (w / v) Polysorbate 80 / 4.0% (w / v) Trehalose (hereafter abbreviated as "LYTR") 10mM Histidine / pH6 / 0.02% (w / v) Polysorbate 80 / 2.0% (w / v) Mannitol (hereafter abbreviated as "LYMA") [Table 11]

[0081] Example 14: Stability test (1) A 10 mg / mL solution of human anti-IL-33 monoclonal antibody (A10-1C04, A23-1A05, A25-2C02, A25-3H04, or A26-1F02) in 10 mM histidine / pH 6 / 0.02% (w / v) Polysorbate 80 / 3.6% (w / v) Sorbitol was prepared and filled into glass vials. The vials were stoppered with halogenated butyl rubber stoppers and stored at 2-8°C for 1, 2, and 3 years for long-term stability testing. Visual inspection was performed under fluorescent light of 2000-3750 lux against a white or black board background for 5 seconds each to confirm the presence or absence of aggregation. Binding activity was assessed as follows: Human IL-33 was added to a 96-well plate and immobilized overnight. After blocking with BSA, sample solution was added to each well, reacted with HRP-labeled anti-human IgG antibody, and color development was performed with TMB. The absorbance at 450 nm and 650 nm was then measured using a plate reader (Molecular Devices) to determine the EC50. The EC50 of the standard solution was similarly determined, and the ratio was calculated. No aggregation or pH drift was observed at any of the measurement points, and the antibody binding activity did not decrease. For example, the results for A10-1C04 ​​are shown in Table 12. [Table 12]

[0082] Example 15: Stability test (2) A 150 mg / mL solution of human anti-IL-33 monoclonal antibody (A10-1C04, A23-1A05, A25-2C02, A25-3H04, or A26-1F02) in 10 mM histidine / pH 6 / 0.02% (w / v) polysorbate 80 / 4.0% (w / v) sorbitol was prepared and filled into glass vials. The vials were stoppered with halogenated butyl rubber stoppers and stored at 2-8°C for 3 and 6 months as a long-term stability test. Evaluation was performed using the same method as in Example 14. Neither aggregation nor pH drift was observed at any measurement point, nor was there a decrease in antibody binding activity. For example, the results for A10-1C04 ​​are shown in Table 13. [Table 13]

Claims

1. A pharmaceutical composition comprising a human anti-IL-33 monoclonal antibody as an active ingredient, the human anti-IL-33 monoclonal antibody has a heavy chain complementarity-determining region 1 (H1), a heavy chain complementarity-determining region 2 (H2), a heavy chain complementarity-determining region 3 (H3), a light chain complementarity-determining region 1 (L1), a light chain complementarity-determining region 2 (L2), and a light chain complementarity-determining region 3 (L3) in a combination of amino acid sequences selected from any one of C1 to C5 in Table 1; A pharmaceutical composition, wherein said pharmaceutical composition is essentially free of sodium chloride or contains less than 30 mM sodium chloride. 【Table 1】

2. The pharmaceutical composition according to claim 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. 【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 is essentially free of sodium chloride.

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

7. The pharmaceutical composition according to any one of claims 1 to 6, which has a pH adjusted to 5 or more and 7 or less.

8. 8. The pharmaceutical composition according to claim 1, wherein the pH is adjusted with a buffer selected from the group consisting of acetate, histidine, and phosphate.

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

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

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

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

16. 16. The pharmaceutical composition of claim 15, wherein the surfactant is a nonionic surfactant.

17. 17. The pharmaceutical composition of 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, comprising 10 mM histidine, 4% (w / v) sorbitol, 0.02% (w / v) polysorbate 80, and 150 mg / ml of the active ingredient, and having a pH adjusted to 5.5 to 6.

5.

19. 19. The pharmaceutical composition of claim 18 for subcutaneous administration.

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

5.

21. 21. The pharmaceutical composition of claim 20 for intravenous administration.

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

23. A freeze-dried 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, the human anti-IL-33 monoclonal antibody has a heavy chain complementarity-determining region 1 (H1), a heavy chain complementarity-determining region 2 (H2), a heavy chain complementarity-determining region 3 (H3), a light chain complementarity-determining region 1 (L1), a light chain complementarity-determining region 2 (L2), and a light chain complementarity-determining region 3 (L3) in a combination of amino acid sequences selected from any one of C1 to C5 in Table 1; The pharmaceutical composition comprises a buffer, a nonionic surfactant, and a polyol. 【Table 3】

Images