Pharmaceutical composition containing a 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 particularly relates to an antibody-containing pharmaceutical composition that suppresses turbidity and improves storage stability such as pH.

Background Art

[0002] In recent years, various antibody-containing pharmaceutical compositions have been developed and put into practical use. Many antibody-containing pharmaceutical compositions are used as intravenous injection pharmaceutical compositions. On the other hand, due to the needs in the medical field, there is an increasing demand to develop an antibody-containing pharmaceutical composition that can be self-injected as a subcutaneous injection pharmaceutical composition.

[0003] When designing a subcutaneous injection antibody-containing pharmaceutical composition, while the antibody dose per administration is large (about 100 to 200 mg), there is generally a limit on the injection volume in subcutaneous injection. Therefore, it is necessary to increase the concentration of the antibody in the administration solution. Thus, a high-concentration antibody-containing pharmaceutical composition using a so-called lyophilization concentration technique, in which a lyophilized pharmaceutical composition is redissolved using less water than before lyophilization to prepare a high-concentration antibody-containing pharmaceutical composition, is often used.

[0004] IgG antibody, which is one of the antibody subtypes and widely used in pharmaceuticals, is a high-molecular-weight glycoprotein of about 150 kDa. Since the structures (amino acid sequences and sugar chain structures) of the variable regions of antibodies are very diverse, different physical properties are exhibited for each antibody molecular species.

[0005] Antibodies may aggregate, decompose, or undergo chemical reactions and denature due to physical stimuli such as heating and vibration, interaction with compounds such as surfactants, oxidizing and reducing agents, and sugars, or long-term storage. When an antibody denatures, there is a concern that the binding ability to an antigen or Fc receptor changes, resulting in a decrease in the function and effect of the antibody, or that the aggregated antibody causes inflammation. The ease of denaturation such as aggregability varies for each antibody molecular species.

[0006] Since antibodies contain charged amino acids, highly polar amino acids, and sugar chains, they may interact with ions in the antibody solution, resulting in an uneven distribution of ions and causing a change in the pH of the antibody solution (pH drift) (Donnan effect). The change in the pH of the antibody solution may affect the storage stability of the antibody, but the strength of the Donnan effect varies for different molecular species of antibodies.

[0007] High-concentration antibody solutions tend to form solutions with high viscosities due to the properties of proteins as macromolecules and intermolecular interactions. When 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 for different molecular species of antibodies.

[0008] Furthermore, when storing antibody solutions for a long time, degradation phenomena such as changes in pH and the formation of insoluble and / or soluble aggregates may become problems, but these also vary for different molecular species of antibodies.

[0009] Generally, various measures have been taken to make pharmaceutical compositions containing antibodies into stabilized pharmaceutical compositions with little loss of the active ingredient even after long-term storage. The active ingredient, the antibody, and various additives such as buffers are dissolved to produce a pharmaceutical composition. However, particularly in pharmaceutical compositions containing high concentrations of antibodies, the technology for preventing antibody aggregation, turbidity, viscosity increase, and pH drift is still insufficient.

[0010] We have obtained multiple human anti-IL-33 monoclonal antibodies that bind to IL-33 (Patent Document 1: International Publication No. WO2015 / 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

Summary of the Invention

Problems 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 Problems

[0013] The present inventors found that when formulating a human anti-IL-33 monoclonal antibody, it becomes turbid, and found that the cause of the turbidity is the formation of aggregates. They also found that the turbidity depends on the NaCl concentration and pH. The present invention has been completed based on such findings.

[0014] That is, the present invention provides the following. [1] A pharmaceutical composition containing 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, and the pharmaceutical composition essentially does not contain sodium chloride or contains less than 30 mM of sodium chloride, a pharmaceutical composition.

Table 1

Table 2

[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 according to

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

[13] The pharmaceutical composition according to

[12] or [12-1], wherein the polyol is 3-5% (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 according to

[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], comprising 10 mM histidine, 4% (w / v) sorbitol, 0.02% (w / v) polysorbate 80, and 150 mg / ml of an active ingredient, and having 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], comprising 10 mM histidine, 3.6% (w / v) sorbitol, 0.02% (w / v) polysorbate 80, and 10 mg / ml of an active ingredient, and having a pH adjusted to 5.5 to 6.5.

[19] The pharmaceutical composition according to

[18] for intravenous administration.

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

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

[21] A lyophilized preparation of the pharmaceutical composition according to any one of [1] to

[20] .

[0015]

[22] A method for treating or preventing an IL-33-related disease, comprising: administering to a subject in need thereof a pharmaceutical composition containing 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 above, and the pharmaceutical composition essentially does not contain sodium chloride or contains less than 30 mM sodium chloride.

[23] The method according to

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

[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 essentially does not contain sodium chloride.

[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.

[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.

[29] The method according to any one of

[22] to

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

[30] The method according to any one of

[22] to

[29] , wherein the pH of the pharmaceutical composition is 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] A method according to any one of

[22] to

[36] , comprising 10 mM histidine, 4% (w / v) sorbitol, 0.02% (w / v) polysorbate 80, and 150 mg / ml of an active ingredient, with the pH adjusted to 5.5 - 6.5.

[38] The method according to

[37] , wherein the administration is subcutaneous administration.

[39] A method according to any one of

[22] to

[36] , comprising 10 mM histidine, 3.6% (w / v) sorbitol, 0.02% (w / v) polysorbate 80, and 10 mg / ml of an active ingredient, with the pH adjusted to 5.5 - 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 the treatment or prevention of an IL - 33 - related disease, comprising a human anti - IL - 33 monoclonal antibody as an active ingredient, wherein the combination of the amino acid sequences of the 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 above, and wherein the pharmaceutical composition essentially does not contain sodium chloride or contains less than 30 mM of sodium chloride.

[0017]

[43] Use for the manufacture of a pharmaceutical composition for the treatment or prevention of an IL - 33 - related disease, wherein the pharmaceutical composition contains a human anti - IL - 33 monoclonal antibody and essentially does not contain sodium chloride or contains less than 30 mM of sodium chloride. Use, 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.

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

[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] , containing 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 containing 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, and the pharmaceutical composition contains 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 essentially does not contain sodium chloride or contains less than 30 mM of sodium chloride. [Advantages of the Invention]

[0018] The pharmaceutical composition containing the 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 excellent in safety and efficacy because turbidity is suppressed. Further, the pharmaceutical composition of the present invention can be stably stored for a long time without significant fluctuation in pH in a solution state, and is suitable for storage and use of the preparation. [Brief Description of the Drawings]

[0019]

Figure 1

Figure 2

[0020] Hereinafter, in order to facilitate understanding of the present invention, terms will be explained.

[0021] [Pharmaceutical Composition] In the present specification, the pharmaceutical composition refers to a composition prepared so as to be administrable to animals such as humans. Therefore, the pharmaceutical composition may refer to a formulated preparation.

[0022] [Essentially] As used herein, "substantially free of sodium chloride" means a case where sodium chloride is not added to the pharmaceutical composition of the present invention, etc., and means a composition that does not contain sodium chloride to the extent that the pharmaceutical composition of the present invention becomes turbid.

[0023] [Surfactant] As used herein, the term "surfactant" generally refers to a substance having a water-friendly moiety (hydrophilic group) and an oil-friendly moiety (lipophilic group / hydrophobic group) within the molecule. Any surfactant generally used in antibody preparations can be used, and examples include 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. Also, the "antibody" as used herein may be a multispecific antibody (e.g., bispecific antibody), a drug conjugate antibody (ADC), and may further be an antigen-binding fragment such as dAbs, scFv, Fab, F(ab)'2, Fab'.

[0025] [Monoclonal antibody] As used herein, the term "monoclonal antibody" 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 such as glycosylation or amino acid modifications. In contrast to polyclonal antibodies, which typically contain different antibodies, each monoclonal antibody binds to a single epitope on the antigen. The modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies and should not be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies used in the present invention may be made by the hybridoma method first described by Kohler et al., Nature, 256:495 (1975), or may be made by recombinant DNA methods (see, e.g., U.S. Patent No. 4,816,567). "Monoclonal antibodies" may also be isolated from phage antibody libraries using, for example, the techniques described in Clackson et al., Nature, 352:624-628 (1991) and Marks et al., J. Mol. Biol., 222:581-597 (1991).

[0026] [Five human anti-IL-33 monoclonal antibodies] The five human anti-IL-33 monoclonal antibodies used in the present invention refer to the five clones of human anti-IL-33 monoclonal antibodies, A10-1C04, A23-1A05, A25-2C02, A25-3H04, and A26-1F02, disclosed in International Publication No. WO 2015 / 099175, and refer to human anti-IL-33 monoclonal antibodies consisting of the heavy and light chains of the following amino acid sequences. (a) A10-1C04: · Heavy chain: EVQLLESGGGLVQPGGSLRLSCAASGFTFSDYYMNWVRQAPGKGLEWVSSISRYSSYIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDIGGMDVWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (Sequence number 1 in the sequence listing); and · Light chain: QSVLTQPPSASGTPGQRVTISCTGSSSNIGAVYDVHWYQQLPGTAPKLLIYRNNQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCQTYDSSRWVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (Sequence number 2 in the sequence listing) (b) A23-1A05: · Heavy chain: EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMHWVRQAPGKGLEWVSSISARSRYHYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARLATRHNAFDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (Sequence number 3 in the sequence listing); and · Light chain: QSVLTQPPSASGTPGQRVTISCSGSSSNIGNNAVSWYQQLPGTAPKLLIYASNMRVIGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDDSQKALVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (Sequence number 4 in the sequence listing) (c) A25-2C02: · Heavy chain: EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMHWVRQAPGKGLEWVSSISARSSYIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARLATRNNAFDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (Sequence number 5 in the sequence listing); and · Light chain: QSVLTQPPSASGTPGQRVTISCSGSSSNIGRNAVNWYQQLPGTAPKLLIYASNMRVSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCWAWDDSQKVGVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (Sequence number 6 in the sequence listing) (d) A25-3H04: · Heavy chain: EVQLLESGGGLVQPGGSLRLSCAASGFTFSRYYMHWVRQAPGKGLEWVSSISAQSSHIYYADSVEGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARLATRQNAFDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (Sequence number 7 in the sequence listing); and · Light chain: QSVLTQPPSASGTPGQRVTISCSGSSSNIGRNAVNWYQQLPGTAPKLLIYASNMRRSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCSAWDDSQKVVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (Sequence number 8 in the sequence listing) (e) A26-1F02: · Heavy chain: EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMHWVRQAPGKGLEWVSSISARSSYLYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARLATRHVAFDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (Sequence number 9 in the sequence listing); and · Light chain: QSVLTQPPSASGTPGQRVTISCSGSSSNIGNNAVNWYQQLPGTAPKLLIYASNMRRPGVPDRFSGSKSGTSASLAISGLRSEDEADYYCEAWDDSQKAVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (Sequence number 10 in the sequence listing)

[0027] [Complementary determining region (CDR)] As used herein, the complementarity-determining region (CDR) refers to the amino acid residues of an antibody that are involved in antigen binding. CDRs are generally also referred to as "hypervariable regions" and are unique amino acid sequences for each molecular species of antibody, 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)). There are 3 CDRs (L1, L2, L3) in the light chain of the antibody and 3 CDRs (H1, H2, H3) in the heavy chain of the antibody. The CDRs of each of the 5 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 (Sequence number 24 in the sequence listing) L1: SGSSSNIGRNAVN (Sequence number 25 in the sequence listing) L2: ASNMRVS (Sequence number 26 in the sequence listing) L3: WAWDDSQKVGV (Sequence number 27 in the sequence listing) (d) A25-3H04: H1: RYYMH (Sequence number 28 in the sequence listing) H2: SISAQSSHIYYADSVEG (Sequence number 29 in the sequence listing) H3: LATRQNAFDI (Sequence number 30 in the sequence listing) L1: SGSSSNIGRNAVN (Sequence number 25 in the sequence listing) L2: ASNMRRS (Sequence number 31 in the sequence listing) L3: SAWDDSQKVVV (Sequence number 32 in the sequence listing) (e) A26-1F02: H1: NYYMH (Sequence number 17 in the sequence listing) H2: SISARSSYLYYADSVKG (Sequence number 33 in the sequence listing) H3: LATRHVAFDI (Sequence number 34 in the sequence listing) L1: SGSSSNIGNNAVN (Sequence number 35 in the sequence listing) L2: ASNMRRP (Sequence number 36 in the sequence listing) L3: EAWDDSQKAVV (Sequence number 37 in the sequence listing)

[0028] [Variable region] In this specification, the variable region refers to the part other than the constant region of a monoclonal antibody, which is involved in the binding to an antigen and determines the specificity of the antibody that varies variously depending on the antigen. There are a heavy chain variable region and a light chain variable region in the variable region. The heavy chain variable regions and light chain variable regions of the 5 clones of A10-1C04, A23-1A05, A25-2C02, A25-3H04, and A26-1F02 are as follows. (a) A10-1C04: · Heavy chain variable region: EVQLLESGGGLVQPGGSLRLSCAASGFTFSDYYMNWVRQAPGKGLEWVSSISRYSSYIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDIGGMDVWGQGTLVTVSS (Sequence No. 38 of the Sequence Listing) · Light chain variable region: QSVLTQPPSASGTPGQRVTISCTGSSSNIGAVYDVHWYQQLPGTAPKLLIYRNNQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCQTYDSSRWVFGGGTKLTVLG (Sequence No. 39 of the Sequence Listing) (b) A23-1A05: · Heavy chain variable region: EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMHWVRQAPGKGLEWVSSISARSRYHYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARLATRHNAFDIWGQGTLVTVSS (Sequence No. 40 of the Sequence Listing) · Light chain variable region: QSVLTQPPSASGTPGQRVTISCSGSSSNIGNNAVSWYQQLPGTAPKLLIYASNMRVIGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDDSQKALVFGGGTKLTVLG (Sequence No. 41 of the Sequence Listing) (c) A25-2C02: · Heavy chain variable region: EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMHWVRQAPGKGLEWVSSISARSSYIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARLATRNNAFDIWGQGTLVTVSS (Sequence No. 42 of the Sequence Listing) · Light chain variable region: QSVLTQPPSASGTPGQRVTISCSGSSSNIGRNAVNWYQQLPGTAPKLLIYASNMRVSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCWAWDDSQKVGVFGGGTKLTVLG (SEQ ID NO: 43 in the Sequence Listing) (d) A25 - 3H04: · Heavy chain variable region: EVQLLESGGGLVQPGGSLRLSCAASGFTFSRYYMHWVRQAPGKGLEWVSSISAQSSHIYYADSVEGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARLATRQNAFDIWGQGTLVTVSS (SEQ ID NO: 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] "Stable" 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) even 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, N.Y., Pubs. (1991) and Jones, A. Adv. Drug Delivery Rev. 10:29-90 (1993). The stability of an antibody-containing pharmaceutical composition can be evaluated for a selected time at a selected temperature. A "stable" antibody-containing pharmaceutical composition is one in which no significant change is observed for 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 for at least 3 months, preferably 6 months, and more preferably 1 year at room temperature (23-27°C), or for 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, such as visual inspection abnormalities (turbidity, etc.), pH, viscosity, binding of the antibody to the antigen, inhibitory activity of the antibody against the antigen molecule (e.g., inhibitory activity of the induction of IL-6 production by IL-33), effector function of the antibody, and degradation of the antibody, can be used as indicators.

[0030] "Effector function" The "effector function" of an antibody refers to those biological activities that are attributable to the Fc region of the antibody (Fc region of the native sequence or Fc region of an amino acid sequence variant). Examples of antibody effector functions 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.

[0031] [Turbidity] In this specification, "cloudiness" refers to a state of being turbid white as determined by visual inspection of color and / or transparency (turbidity). Whether there is cloudiness can be further analyzed by measuring fine particles using a flow cytometric particle image analyzer or a particle counter, measuring interaction parameters (hereinafter simply referred to as "interaction parameters" or "Kd values") by dynamic light scattering (DLS), and measuring turbidity (absorbance at 650 nm (OD650)). As analysis values correlated with "cloudiness", for example, the OD650 of an antibody-containing pharmaceutical composition 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 by a particle counter is 500, 750, 1000, 1250, or 1500 particles / mL or more.

[0032] [Lyophilized preparation] In this specification, "lyophilized preparation" refers to a dry (e.g., freeze-dried) pharmaceutical composition with little water. The lyophilization technique of antibodies is 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 called NF-HEV. When IL-33 is released extracellularly as a cytokine, it binds to the IL-33 receptor (ST2 and IL-1RAcP) and has the function of initiating intracellular signal transduction in cells expressing the IL-33 receptor. Signal transduction induced by IL-33 includes, but is not limited to, the NF-κB pathway and the MAPKKs pathway, and ultimately induces 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 are 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. Cytokines, chemokines, and inflammatory mediators induced by IL-33 are involved in the migration, cytokine production, and degranulation of immune system cells and cause inflammation. In the present invention, "five human anti-IL-33 monoclonal antibodies" may refer to either full-length IL-33 or mature IL-33 as long as they bind to inhibit at least one of the above functions, or may be homologous derivatives or mutants thereof. Also, it may be human IL-33 or IL-33 derived from other organisms.

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

[0035] [Isotonic] As used herein, an "isotonic" formulation has substantially the same osmotic pressure as human blood. Isotonic pharmaceutical compositions generally have an osmotic pressure ratio of about 0.9 to 1.2 based on blood. Osmotic pressure can be measured, for example, using a vapor pressure type or freezing point type osmometer.

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

[0037] 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 only to those embodiments.

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

[0039] 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. Further, after making it into a solution containing a desired buffer, it is also possible to add saccharides, surfactants, etc. later for formulation.

[0040] The present inventors have found that when five types of human anti-IL-33 monoclonal antibodies are made into a solution, they partially aggregate and become turbid. The turbidity is due to the formation of fine particles which are aggregates containing the antibody. When used as a pharmaceutical composition, there are concerns about a decrease in the biological activity of the antibody as the active ingredient, a deterioration in pharmacokinetics due to the induction of anti-drug antibodies (ADA) by aggregates with high immunogenicity, and further the induction of inflammation 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 contains five types of human anti-IL-33 monoclonal antibodies as an active ingredient, and may further contain salts, buffers, surfactants, saccharides, etc.

[0041] [Salt concentration] The inventors of the present invention have found that the main cause of the turbidity of five types of human anti-IL-33 monoclonal antibodies is mainly salt. Therefore, the pharmaceutical composition according to 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, and more preferably contains essentially no salt. Also, the pharmaceutical composition according to 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, and more preferably contains essentially no salt. In the pharmaceutical composition according to the present invention, examples of the low-concentration or essentially non-containing salt include inorganic salts or organic salts. Examples of inorganic salts include, in particular, sodium chloride, potassium chloride, magnesium chloride, calcium chloride, sodium sulfate, potassium sulfate, magnesium sulfate, calcium sulfate, but sodium chloride is preferred.

[0042] [Buffer and pH] The pH of the pharmaceutical composition of the present invention is adjusted by a buffer. Examples of buffers suitable as pharmaceutical compositions include, without limitation, gluconates, histidine, citrates, phosphates [e.g., sodium or potassium], succinates [e.g., sodium], acetates, tris(hydroxymethyl)aminomethane, glycine, arginine, and combinations thereof, which can be used appropriately depending on the pH value to be adjusted. The pharmaceutical composition of the present invention preferably contains acetate, histidine, or phosphate as a buffer, and more preferably contains histidine as a buffer. The concentration of the buffer is not particularly limited as long as it is pharmaceutically 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. Also, 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 an inorganic salt, it is preferable to use a concentration of less than 30 mM, especially 10 mM or less.

[0043] For the pharmaceutical composition of the present invention, histidine as a buffer (for example, at a concentration of 5 mM to 50 mM, for example, 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 may be within the range of 4.0 to 8.0, a pH within the range of 4.5 to 7.5, for example, 5.0 to 7.0, 5.2 to 6.8, for example, about 4.2, about 4.3, about 4.4, about 4.5, about 4.6, about 4.7, about 4.8, about 4.9, about 5.0, about 5.1, about 5.2, about 5.3, about 5.4, about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, about 6.0, about 6.1, about 6.2, about 6.3, about 6.4, about 6.5, about 6.6, about 6.7, about 6.8, about 6.9, about 7.0, about 7.1, about 7.2, about 7.3, about 7.4, about 7.5, about 7.6, about 7.7, about 7.8 are common. The pharmaceutical composition of the present invention shows an increase in turbidity during long-term storage at pH 4 or pH 8. Also, the pharmaceutical composition of the present invention shows significant pH drift during long-term storage at pH 4 or pH 8. 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.

[0044] [Surfactant] The pharmaceutical composition of the present invention preferably contains a surfactant. Surfactants suitable as pharmaceutical compositions include, but are not limited to, nonionic surfactants, ionic surfactants, zwitterionic surfactants, and combinations thereof. General 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 monolinoerate), 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 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 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 stearic acid amide), C10-C18 alkyl sulfuric acid (for example, sodium cetyl sulfate, sodium lauryl sulfate, sodium oleyl sulfate), polyoxyethylene C10-C18 alkyl ether sulfuric acid with an additional average of 2-4 moles of ethylene oxide units (for example, sodium polyoxyethylene lauryl sulfate) and C1-C18 alkyl sulfosuccinate esters (for example, sodium lauryl sulfosuccinate ester), as well as natural surfactants such as lecithin, glycerophospholipids, sphingophospholipids (for example, sphingomyelin) and sucrose esters of C12-C18 fatty acids are included., 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 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, and as an example, it can be used at a concentration of about 0.01% (w / v) to about 0.1% (w / v), for example, about 0.01% (w / v) to about 0.04% (w / v), for example, about 0.01% (w / v), about 0.02% (w / v), about 0.04% (w / v), about 0.06% (w / v), about 0.08% (w / v), about 0.1% (w / v). Among the surfactants, polysorbate 80 (Tween 80) is particularly beneficial. In one embodiment, the stable pharmaceutical composition contains about 0.02% (w / v) of polysorbate 80. In one embodiment, the stable pharmaceutical composition contains about 0.02% (w / v) of polysorbate 20.

[0045] [Polyol] The pharmaceutical composition of the present invention preferably contains a polyol. By adding the polyol, the osmotic pressure of the pharmaceutical composition can be adjusted, and the formation of aggregates can be suppressed. 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 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) is preferred. A polyol, also referred to as a polyhydric alcohol, may be any molecule having two or more alcoholic hydroxyl groups. Examples include glycerol (glycerin), propylene glycol, polyethylene glycol (PEG), saccharides, etc., and saccharides are preferably included. Saccharides suitable as the pharmaceutical composition of the present invention include, without limitation, compounds having the general formula (CH2O)n including monosaccharides, disaccharides, trisaccharides, polysaccharides, sugar alcohols, reducing sugars, non-reducing sugars, etc., and derivatives thereof. Examples of saccharides include, as monosaccharides, glucose, fructose, galactose, as disaccharides, sucrose, trehalose, lactose, maltose, lactulose, maltulose, iso-maltulose, melibiose, as trisaccharides, melezitose, raffinose, maltotriose, as polysaccharides, stachyose, dextran, as sugar alcohols, sorbitol, mannitol, erythritol, maltitol, lactitol, arabitol, xylitol, etc. Among monosaccharides, disaccharides, and trisaccharides, reducing sugars include glucose, fructose, lactose, maltose, lactulose, maltulose, iso-maltulose, melibiose, melezitose, maltotriose, and non-reducing sugars include trehalose, sucrose, raffinose. Saccharides included in the pharmaceutical composition of the present invention are preferably sorbitol, sucrose, trehalose or mannitol, and sorbitol or sucrose is most preferred. The concentration of the saccharide included 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. Also, concentrations of the saccharide of 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 are preferred. In one embodiment, a stable pharmaceutical composition contains 3% to 5% (w / v) (165 to 275 mM) of sorbitol. In one embodiment, a stable pharmaceutical composition contains about 3.6% (w / v) or about 4% (w / v) (about 200 mM or about 220 mM, respectively) of sorbitol.

[0046] Preferably, the osmotic pressure ratio of the pharmaceutical composition of the present invention is 0.5 to 4, more preferably 0.7 to 3, still 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, about 2.0. The osmotic pressure of the pharmaceutical composition can be adjusted by the concentration of components added in addition to the active ingredient, such as salts or polyols. From the perspective of reducing the salt concentration, it is preferable to adjust the osmotic pressure of the pharmaceutical composition using polyols.

[0047] [Human anti-IL-33 monoclonal antibody] The pharmaceutical composition of the present invention contains a human anti-IL-33 monoclonal antibody (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02) as an active ingredient. The concentration of the human anti-IL-33 monoclonal antibody contained in the pharmaceutical composition of the present invention is not particularly limited as long as it is pharmaceutically acceptable, but is preferably 1 mg / mL to 200 mg / mL, 5 mg / mL to 175 mg / mL, 10 mg / mL to 150 mg / ml, 20 mg / mL to 150 mg / mL, for example, about 1, about 5, about 10, about 20, about 25, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, about 150, about 160, about 170, about 175 mg / mL. Since the viscosity is high when the antibody concentration of the pharmaceutical composition of the present invention is 175 mg / mL or more, 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.

[0048] The pharmaceutical composition of the present invention preferably has an appropriate viscosity and can be easily administered to any patient. If the viscosity is low, it can be administered without requiring strong force. The inventors have found that for the pharmaceutical composition of the present invention, when the viscosity is about 20 cP or more, injection from a syringe during administration becomes somewhat difficult. Therefore, the pharmaceutical composition according to the present invention preferably has a viscosity of 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 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 by rheometry (rotational or capillary).

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

[0050] Examples of IL-33-related diseases include, but are not limited to, asthma, atopic dermatitis, urticaria, allergic rhinitis, anaphylactic shock, eosinophilic rhinosinusitis, eosinophilia syndrome, Churg-Strauss syndrome, allergic encephalomyelitis, rheumatoid polymyalgia, rheumatic heart disease, multiple sclerosis, arthritis (e.g., rheumatoid arthritis, juvenile arthritis, psoriatic arthritis, osteoarthritis, Reiter's syndrome, etc.), systemic lupus erythematosus (including discoid lupus), psoriasis, ankylosing spondylitis, hepatitis (e.g., autoimmune hepatitis, chronic active hepatitis, etc.), inflammatory bowel disease (e.g., ulcerative colitis, Crohn's disease, gluten-sensitive enteropathy, etc.), systemic lupus erythematosus, Sjogren's syndrome, Behcet'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 drug resistance, alopecia greata, antiphospholipid syndrome, autoimmune diseases of the adrenal gland (e.g., autoimmune Addison's disease, etc.), celiac sprue-dermatitis, chronic fatigue immune dysfunction syndrome (CFIDS), cold agglutinin disease, essential 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, stiff-man syndrome, vitiligo, sarcoidosis, polyendocrine disorder, other endocrine insufficiency, arteriosclerosis, liver fibrosis (e.g., primary biliary cirrhosis, etc.), pulmonary fibrosis (e.g., idiopathic pulmonary fibrosis, etc.), chronic obstructive pulmonary disease, scleroderma (including CREST syndrome, Raynaud's phenomenon, etc.), endometriosis, adenomyosis, tubulointerstitial nephritis, dense deposit disease, acute kidney injury, myocarditis, cardiomyopathy, neuritis (e.g., Guillain-Barré syndrome, etc.), polyarteritis nodosa, cardiotomy syndrome, chronic inflammatory demyelinating polyneuropathy, IgA neuropathy, lichen planus, Meniere's disease, post-myocardial infarction (post-MI), uveitis, uveitis ophthalmiaOpthalmia, vasculitis, primary agammaglobulinemia, cancer (e.g., brain tumor, laryngeal cancer, oral and lip 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 myelogenous leukemia, Ewing's tumor, Hodgkin's disease, non-Hodgkin lymphoma, melanoma, mesothelioma, multiple myeloma, etc.), infections resistant to elimination by the immune system (e.g., severe acute respiratory syndrome (SARS)), lethal cytokine storms associated with highly virulent influenza infections, and sepsis. Preferably, asthma, atopic dermatitis, allergic rhinitis, 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.

[0051] The "stable" pharmaceutical composition of the present invention shows no significant changes at refrigeration temperature (2 - 8°C) for at least 12 months, preferably 2 years, more preferably 3 years, or at room temperature (22 - 28°C) for at least 3 months, preferably 6 months, more preferably 1 year. For example, after storage 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, and most preferably 500 particles / mL or less.

[0052] To the pharmaceutical composition of the present invention, preservatives, anti-adsorption agents, soothing agents, sulfur-containing reducing agents, antioxidants, etc. can be appropriately added as needed.

[0053] The preservative is not particularly limited as long as it is pharmaceutically acceptable, and examples thereof include methyl paraoxybenzoate, ethyl paraoxybenzoate, sorbic acid, phenol, cresol, and chlorocresol.

[0054] The anti-adsorption agent is not particularly limited as long as it is pharmaceutically acceptable, and examples thereof include human serum albumin, lecithin, dextran, ethylene oxide-propylene oxide copolymer, hydroxypropyl cellulose, methyl cellulose, polyoxyethylene hydrogenated castor oil, and polyethylene glycol.

[0055] The pain reliever 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 N-acetylcysteine, N-acetylhomocysteine, thioctic acid, thiodiglycol, thioethanolamine, thioglycerol, thiosorbitol, thioglycolic acid and its salts, sodium thiosulfate, glutathione, and those having a sulfhydryl group such as 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-ascorbic acid palmitate, L-ascorbic acid stearate, sodium bisulfite, sodium sulfite, triamyl gallate, propyl gallate, or 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 preparation for longer-term 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 only to these.

Example

[0060] Comparative Example 1: Turbidity 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) Polysorbate 80) (hereinafter abbreviated as "P7N") to a concentration of 150 mg / mL, and turbidity was observed when visually checking the physical properties (for example, shown in FIG. 1 for A10-1C04). The presence or absence of turbidity was confirmed visually under white light (13W fluorescent lamp) against a black background. Also, when this pharmaceutical composition was measured for sub-visible particles (particles of 5 μm or more) with a FlowCam (manufactured by Fluid Imaging Technologies), 1,919 particles / mL were detected. From these results, it was considered that human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02) had high aggregability and were difficult to formulate.

[0061] Comparative Example 2: Effect of pH on Turbidity of Pharmaceutical Composition Containing Human Anti-IL-33 Monoclonal Antibody 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 follows based on the formulation of Comparative Example 1 to change the pH of the pharmaceutical composition. The antibody concentration was 150 mg / ml. The examined formulations are shown below. ·10 mM Na-acetate / pH 4 / 150 mM NaCl / 0.02% (w / v) Polysorbate 80 (hereinafter abbreviated as "A4N") ·10 mM Na-acetate / pH 5 / 150 mM NaCl / 0.02% (w / v) Polysorbate 80 (hereinafter abbreviated as "A5N") · 10 mM Histidine / pH 6 / 150 mM NaCl / 0.02% (w / v) Polysorbate 80 (hereinafter abbreviated as "H6N"). · 10 mM Na-phosphate / pH 6 / 150 mM NaCl / 0.02% (w / v) Polysorbate 80 (hereinafter abbreviated as "P6N"). · 10 mM Na-phosphate / pH 7 / 150 mM NaCl / 0.02% (w / v) Polysorbate 80 (hereinafter abbreviated as "P7N") and · 10 mM Na-phosphate / pH 8 / 150 mM NaCl / 0.02% (w / v) Polysorbate 80 (hereinafter abbreviated as "P8N"). As a result of evaluating the properties, turbidity occurred in all formulations at pH 4, 5, 6, 7, and 8. From this result, it was considered that changing the pH of the formulation did not improve the turbidity. Note that the visual confirmation of the properties was observed by the method described in Comparative Example 1.

[0062] Example 1: Turbidity Suppression Effect by Sugar Addition To eliminate the turbidity of the human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04, or A26-1F02), the following formulations were evaluated by adding 5% sorbitol instead of sodium chloride contained in A4N, A5N, H6N, P6N, P7N, and P8N, which are the formulations 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) Polysorbate 80 (hereinafter abbreviated as "A4S"). · 10 mM Na-acetate / pH 5 / 5% (w / v) Sorbitol / 0.02% (w / v) Polysorbate 80 (hereinafter abbreviated as "A5S"). · 10 mM Histidine / pH 6 / 5% (w / v) Sorbitol / 0.02% (w / v) Polysorbate 80 (hereinafter abbreviated as "H6S"). · 10 mM Na - phosphate / pH6 / 5% (w / v) Sorbitol / 0.02% (w / v) Polysorbate 80 (hereinafter abbreviated as "P6S"). · 10 mM Na - phosphate / pH7 / 5% (w / v) Sorbitol / 0.02% (w / v) Polysorbate 80 (hereinafter abbreviated as "P7S"). · 10 mM Na - phosphate / pH8 / 5% (w / v) Sorbitol / 0.02% (w / v) Polysorbate 80 (hereinafter abbreviated as "P8S"). As a result of evaluating the properties and OD650 (turbidity), turbidity was improved by adding sorbitol instead of sodium chloride (shown in Figure 2 for A10 - 1C04, for example). Also, by adding sorbitol, the increase in OD650 was suppressed even after 3 months at 40°C. In particular, the inhibitory effect was good at pH 5 - 7 (Table 3). Note that the visual confirmation of the properties was observed by the method described in Comparative Example 1. OD650 was measured at 650 nm for 100 μL of the solution of each formulation using a Molecular Devices 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 a buffer of 10 mM Histidine / pH 6.0 or pH 5.5, and the interaction parameter (Kd value), which is an index of aggregability, was measured when 0, 50, or 100 mM of NaCl was added. The measurement temperature was 25 °C, and the antibody concentrations were adjusted to 0.5, 1, 2.5, 5, 10, and 20 mg / mL. As a result, since the Kd value became negative (aggregability increased) when the NaCl addition amount was 50 mM or more, it was considered that the NaCl addition amount was preferably less than 50 mM (for example, as shown in Table 4 for A10-1C04). The Kd value can be calculated by obtaining the diffusion coefficient (Dm) by the dynamic light scattering method, obtaining the slope from the plot of the antibody concentration (horizontal axis) and the diffusion coefficient (vertical axis), and dividing this slope by the diffusion coefficient (D0) at a concentration of 0. That is, the Kd value is calculated from the following relational expression. The larger the positive value of the Kd value, the lower the aggregability. Dm = D0(1 + Kd value × [antibody concentration])

[0065]

Table 4

[0066] Example 3: Salt Concentration and Turbidity (2) NaCl was added at 0, 5, 10, and 30 mM to 10 mM Histidine / pH 6.0 / 3.6% (w / v) Sorbitol, and the Kd value, which is an index of aggregability, was measured. The antibody used was A10-1C04 at 2.5, 5, 10, and 14 mg / mL. As a result, since the Kd value became negative when the NaCl addition amount was 30 mM or more (the Kd values for 0, 5, 10, and 30 mM of NaCl were 17.3, 7.3, 1.2, and -4.0 mL / g, respectively), it was considered that the NaCl addition amount was preferably less than 30 mM. The Kd value was calculated by the method described in Example 2.

[0067] Example 4: pH Drift 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 examined (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) (shown in Table 5 for A10-1C04, for example). Therefore, formulations with pH values between 5 and 7, which exhibit less pH drift, were considered favorable.

[0068]

Table 5

[0069] Example 5: Antibody Concentration and Viscosity In a formulation of 10 mM Histidine / pH6 / 3.6% (w / v) Sorbitol / 0.02% (w / v) Polysorbate80, pharmaceutical compositions were prepared with antibody concentrations of 15, 50, 100, 125, 150, 175, and 200 mg / ml using human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04, or A26-1F02). The viscosity at a measurement temperature of 25 °C was measured using a viscometer (DV3TLVCJ0 type viscometer manufactured by Brookfield (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, and it was found that the viscosity increased rapidly when exceeding 150 mg / mL. Furthermore, solutions with each viscosity were prepared, and the injectability using a syringe was examined. It was found that administration became slightly difficult when the viscosity exceeded approximately 20 cP, and an antibody concentration of less than 175 mg / ml was considered favorable.

[0070] Example 6: Sugar Addition and Aggregation Evaluation and Inhibitory Effect on Fine Particle Formation Compositions of 10 mM Histidine / pH 6.0 of human anti-IL-33 monoclonal antibodies (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02) were prepared at antibody concentrations of 0.6125, 1.25, 2.5, 5 and 10 mg / mL, and the effect on the Kd value, which is an index of aggregability when sucrose or sorbitol is added, was confirmed. The Kd value was calculated by the method described in Example 2. For example, the Kd value when using A10-1C04 was 33.4 mL / g without sugar addition, and when 3.6% (w / v) sucrose or sorbitol was added, they were 24.7 and 33.5 mL / g respectively, both of which were positive values. From this result, it was considered that sucrose could be added instead of sorbitol. Also, for the formulation with 3.6% (w / v) sorbitol added at an antibody concentration of 10 mg / ml (10 mM Histidine / pH 6.0 / 3.6% (w / v) Sorbitol), the number of particles of 1.5 μm or more at the time of preparation and after storage at 50°C for 1 week was measured using a particle counter HIAC (System 9703+ type manufactured by HACH). The measurement by HIAC was carried out 4 times for each sample with an injection volume of 100 μL, and the first data was rejected. As a result, compared with the case without sorbitol addition, when 3.6% (w / v) sorbitol was added, the increase in the number of particles was suppressed, so it was considered that adding sorbitol had an effect of suppressing aggregation (shown in Table 6 for A10-1C04, for example).

[0071]

Table 6

[0072] Example 7: Aggregation Inhibitory Effect of Surfactant A composition of 10 mM Histidine / pH6 / 3.6% (w / v) Sorbitol with 10 mg / mL of human anti-IL-33 monoclonal antibody (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02) was prepared, and the effect on aggregation was confirmed when 0.02% (w / v) of polysorbate 20 or polysorbate 80 was added as a surfactant. Aggregation was not observed during preparation regardless of the addition or non-addition of the surfactant.

[0073] Example 8: Stability of Subcutaneous Administration Formulation and Intravenous Administration Formulation An intravenous administration preparation (10 mg / mL antibody / 10 mM Histidine / pH6 / 3.6% (w / v) Sorbitol / 0.02% (w / v) polysorbate 80) and a subcutaneous administration preparation (150 mg / mL antibody / 10 mM Histidine / pH6 / 4% (w / v) Sorbitol / 0.02% (w / v) polysorbate 80) of human anti-IL-33 monoclonal antibody (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02) were prepared. When examined for the intravenous administration preparation, no turbidity, increase in the number of aggregated particles, or pH drift was observed after storage at 5°C for 24 months, and it was confirmed to be stable. The stability of the subcutaneous administration preparation can be evaluated in the same manner.

[0074] Example 9: Effect of pH on Turbidity of Pharmaceutical Composition Containing Human Anti-IL-33 Monoclonal Antibody (2) In order to eliminate the turbidity of the human anti-IL-33 monoclonal antibody (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02), the following formulations without sodium chloride were prepared within the pH range of the formulation confirmed in Comparative Example 2. · 10 mM Na-acetate / pH4 / 0.02% (w / v) Polysorbate80 (hereinafter abbreviated as "A4"). · 10 mM Na-acetate / pH5 / 0.02% (w / v) Polysorbate80 (hereinafter abbreviated as "A5"). · 10 mM Histidine / pH6 / 0.02% (w / v) Polysorbate 80 (hereinafter abbreviated as "H6"). · 10 mM Histidine / pH7 / 0.02% (w / v) Polysorbate 80 (hereinafter abbreviated as "H7"). · 10 mM Na - phosphate / pH8 / 0.02% (w / v) Polysorbate 80 (hereinafter abbreviated as "P8"). For example, as a result of evaluating the properties of 150 mg / mL of A10 - 1C04, turbidity was improved by the non - addition of sodium chloride. Also, particularly, the inhibitory effect was good at pH 4 - 7 (Table 7).

[0075]

Table 7

[0076] Example 10: Effect of Salt on Turbidity of Pharmaceutical Composition Containing Human Anti-IL-33 Monoclonal Antibody (2) 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 A4, A5, and H6, which are the formulations of Example 9, and evaluated. · 10 mM Na - acetate / pH4 / 10 mM NaCl / 0.02% (w / v) Polysorbate 80 (hereinafter abbreviated as "A4N10"). · 10 mM Na - acetate / pH4 / 30 mM NaCl / 0.02% (w / v) Polysorbate 80 (hereinafter abbreviated as "A4N30"). · 10 mM Na - acetate / pH4 / 50 mM NaCl / 0.02% (w / v) Polysorbate 80 (hereinafter abbreviated as "A4N50"). · 10 mM Na - acetate / pH4 / 100 mM NaCl / 0.02% (w / v) Polysorbate 80 (hereinafter abbreviated as "A4N100"). · 10 mM Na - acetate / pH5 / 10 mM NaCl / 0.02% (w / v) Polysorbate 80 (hereinafter abbreviated as "A5N10"). · 10 mM Na-acetate / pH 5 / 30 mM NaCl / 0.02% (w / v) Polysorbate 80 (hereinafter abbreviated as "A5N30"). · 10 mM Na-acetate / pH 5 / 50 mM NaCl / 0.02% (w / v) Polysorbate 80 (hereinafter abbreviated as "A5N50"). · 10 mM Na-acetate / pH 5 / 100 mM NaCl / 0.02% (w / v) Polysorbate 80 (hereinafter abbreviated as "A5N100"). · 10 mM Histidine / pH 6 / 10 mM NaCl / 0.02% (w / v) Polysorbate 80 (hereinafter abbreviated as "H6N10"). · 10 mM Histidine / pH 6 / 30 mM NaCl / 0.02% (w / v) Polysorbate 80 (hereinafter abbreviated as "H6N30"). · 10 mM Histidine / pH 6 / 50 mM NaCl / 0.02% (w / v) Polysorbate 80 (hereinafter abbreviated as "H6N50"). · 10 mM Histidine / pH 6 / 100 mM NaCl / 0.02% (w / v) Polysorbate 80 (hereinafter abbreviated as "H6N100"). For example, as a result of evaluating the properties of 150 mg / mL of A10-1C04, turbidity was suppressed at pH 4 - 6 with 10 mM sodium chloride, and turbidity was observed with sodium chloride exceeding 30 mM (Table 8).

Table 8

[0077] Example 11: Aggregation Inhibitory Effect of Surfactant (2) To confirm 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 with added surfactants were prepared. Subsequently, these specimens were rotated at 30 rpm using a rotator in a constant temperature bath at 50°C. The specimens taken out over time were visually inspected for 5 seconds each against a white board and a black board under a fluorescent lamp with an illuminance of 2000 - 3750 Lux. In the system without added surfactant, a large number of aggregates were observed after 2 days, but no aggregates were observed in the specimens with added polysorbate 80 and poloxamer 188 even after 7 days. In particular, no aggregates were observed even after 14 days in the system with 0.02% or more added polysorbate 80. For the specimens with added polysorbate 20, no foreign substances were observed until 4 days later, a small amount of foreign substances were observed after 7 days, but no foreign substances were observed after 14 days. For example, Table 9 shows the results obtained for A10-1C04. 10 mM Histidine / pH6 (hereinafter abbreviated as 'H6(-)') 10 mM Histidine / pH6 / 0.02% (w / v) Poloxamer188 (hereinafter abbreviated as 'H6PX') 10 mM Histidine / pH6 / 0.02% (w / v) Polysorbate20 (hereinafter abbreviated as 'H6P20') 10 mM Histidine / pH6 / 0.01% (w / v) Polysorbate80 (hereinafter abbreviated as 'H6PS1') 10 mM Histidine / pH6 / 0.02% (w / v) Polysorbate80 (hereinafter abbreviated as 'H6PS2') 10 mM Histidine / pH6 / 0.05% (w / v) Polysorbate80 (hereinafter abbreviated as 'H6PS5')

Table 9

[0078] Example 12: Sugar Addition and Aggregation Evaluation (2) A composition of 10 mM Histidine / pH6.0 of a 10 mg / mL human anti-IL-33 monoclonal antibody (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02) was prepared, and a chemical solution added with mannitol, trehalose, sucrose or sorbitol was prepared. Then, these specimens were stored at -80 °C for 8 hours or more and frozen, and then left at room temperature for 4 hours or more to thaw. These were repeated 6 times. Then, visual inspection was performed for 5 seconds each against a white board and a black board under a fluorescent lamp of 2000 to 3750 Lux. By adding sugar, the amount of aggregates after freeze-thawing was reduced compared to the non-addition system, and particularly, an inhibitory effect on aggregates was observed in the systems using sorbitol and sucrose. For example, Table 10 shows the results obtained for A10-1C04. 10 mM Histidine / pH6 (hereinafter abbreviated as 'H6(-)') 10 mM Histidine / pH6 / 3.0% (w / v) Sorbitol (hereinafter abbreviated as 'H6So3') 10 mM Histidine / pH6 / 3.6% (w / v) Sorbitol (hereinafter abbreviated as 'H6So3.6') 10 mM Histidine / pH6 / 4.0% (w / v) Sorbitol (hereinafter abbreviated as 'H6So4') 10 mM Histidine / pH6 / 5.0% (w / v) Sorbitol (hereinafter abbreviated as 'H6So5') 10 mM Histidine / pH6 / 3.6% (w / v) Sucrose (hereinafter abbreviated as 'H6Su') 10 mM Histidine / pH6 / 3.6% (w / v) Trehalose (hereinafter abbreviated as 'H6Tr') 10 mM Histidine / pH6 / 3.6% (w / v) Mannitol (hereinafter abbreviated as 'H6Ma')

[0079] [Table 10]

[0080] Example 13: Freeze-Drying A composition of 10 mM Histidine / pH6.0 / 0.02% (w / v) Polysorbate 80 of a human anti-IL-33 monoclonal antibody (A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02) at 150 mg / mL was prepared, and after adding mannitol, trehalose, sucrose or sorbitol, it was lyophilized by shelf freeze-drying (manufactured by Kyowa Vacuum). The properties were visually confirmed by the shape of the cake after lyophilization. Redissolution was confirmed by adding water for injection and checking whether the cake shape remained in the solution after standing at 5°C for half a day. The cake shape was good for all specimens, and redissolution after adding water for injection was confirmed. It was confirmed that preparation with a lyophilized preparation was possible. For example, the results obtained for A10-1C04 are shown in Table 11. 10 mM Histidine / pH6 / 0.02% (w / v) Polysorbate 80 / 4.0% (w / v) Sorbitol (hereinafter abbreviated as 'LYSO') 10 mM Histidine / pH6 / 0.02% (w / v) Polysorbate 80 / 4.0% (w / v) Sucrose (hereinafter abbreviated as 'LYSU') 10 mM Histidine / pH6 / 0.02% (w / v) Polysorbate 80 / 4.0% (w / v) Trehalose (hereinafter abbreviated as 'LYTR') 10 mM Histidine / pH6 / 0.02% (w / v) Polysorbate 80 / 2.0% (w / v) Mannitol (hereinafter abbreviated as 'LYMA')

Table 11

[0081] Example 14: Stability Test (1) A composition of 10 mM Histidine / pH6 / 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) was prepared, filled into glass vials, stoppered with a butyl rubber halogenated stopper, and stored at a temperature of 2-8°C for 1 year, 2 years and 3 years as a long-term stability test. Visual inspection was performed for 5 seconds each against a white background and a black background under a fluorescent lamp of 2000-3750 Lux to confirm the presence or absence of aggregation. The binding activity was evaluated by the following method. Human IL-33 was added to a 96-well plate and immobilized overnight. After blocking with BSA, the sample solution was added to each well, reacted with an HRP-labeled anti-human IgG antibody, and developed with TMB. Then, the absorbances at 450 nm and 650 nm were measured with a plate reader (Molecular Devices), and the EC50 was determined. The EC50 of the standard solution measured in the same manner was determined, and the ratio was calculated. Aggregation and pH drift were not observed at any measurement points, and the binding activity of the antibody did not decrease. For example, the results for A10-1C04 are shown in Table 12.

Table 12

[0082] Example 15: Stability Test (2) A composition of 10 mM Histidine / pH6 / 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) was prepared, filled into glass vials, stoppered with a butyl rubber halogenated stopper, and stored at a temperature of 2-8°C for 3 months and 6 months as a long-term stability test. The evaluation was performed in the same manner as in Example 14. Aggregation and pH drift were not observed at any measurement points, and the binding activity of the antibody did not decrease. For example, the results for A10-1C04 are shown in Table 13.

Table 13

Claims

1. A pharmaceutical composition containing a human anti-IL-33 monoclonal antibody as an active ingredient, wherein the human anti-IL-33 monoclonal antibody is A10-1C04, A23-1A05, A25-2C02, A25-3H04 or A26-1F02, the pharmaceutical composition essentially does not contain sodium chloride or contains 10 mM or less of sodium chloride, the pharmaceutical composition is 10 mg / mL to 150 mg / mL of the human anti-IL-33 monoclonal antibody, as a surfactant, 0.01 to 0.1% (w / v) of polysorbate 80, and as a polyol, contains 3 to 5% (w / v) of sorbitol, a pharmaceutical composition whose pH is adjusted to 5 or more and 7 or less with a buffer of acetic acid, histidine or phosphate.

2. The pharmaceutical composition according to claim 1, which essentially does not contain sodium chloride.

3. The pharmaceutical composition according to claim 1 or 2, wherein the pH is adjusted with histidine.

4. The pharmaceutical composition according to any one of claims 1 to 3, which contains 0.02 to 0.05% (w / v) of polysorbate 80 as a surfactant.

5. The pharmaceutical composition according to any one of claims 1 to 4, which contains 5 mM to 20 mM of a buffer.

6. The pharmaceutical composition according to any one of claims 1 to 5, for subcutaneous administration.

7. The pharmaceutical composition according to any one of claims 1 to 5, for intravenous administration.

8. The pharmaceutical composition according to any one of claims 1 to 7, wherein the human anti-IL-33 monoclonal antibody consists of a heavy chain and a light chain of any of the following amino acid sequences: (a) a heavy chain and a light chain of the amino acid sequences of SEQ ID NOs: 1 and 2 in the sequence listing; (b) a heavy chain and a light chain of the amino acid sequences of SEQ ID NOs: 3 and 4 in the sequence listing; (c) a heavy chain and a light chain of the amino acid sequences of SEQ ID NOs: 5 and 6 in the sequence listing; (d) a heavy chain and a light chain of the amino acid sequences of SEQ ID NOs: 7 and 8 in the sequence listing; and (e) a heavy chain and a light chain of the amino acid sequences of SEQ ID NOs: 9 and 10 in the sequence listing.

9. The pharmaceutical composition according to any one of claims 1 to 8, wherein the active ingredient is A10-1C04.

10. The pharmaceutical composition according to claim 9, wherein the A10-1C04 consists of a heavy chain and a light chain of the amino acid sequences of SEQ ID NOs: 1 and 2 in the sequence listing.

11. A lyophilized preparation of the pharmaceutical composition according to any one of claims 1 to 10.

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