Antibody-drug conjugate preparation and lyophilization for the same

Abstract

To provide a pharmaceutical composition (especially in the form of an aqueous injection or a lyophilized injection) for an antibody-drug conjugate where the formation of aggregates and the generation of decomposition products are suppressed, and an efficient method of lyophilizing an aqueous solution to a lyophilized injection.SOLUTION: A pharmaceutical composition comprises (i) an antibody-drug conjugate represented by the following formula (where A represents a connecting position to an antibody), (ii) a histidine buffer, (iii) sucrose or trehalose, and (iv) a surfactant.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to the formulation of certain antibody-drug conjugates and methods for lyophilization thereof. [Background technology]

[0002] The antibody that binds to an antigen that is expressed on the surface of cancer cells and can be internalized into the cells has cytotoxicity. Antibody-Drug Conjugates ADCs (antibody-derived jugates) are a promising candidate for the treatment of cancer by selectively delivering drugs to cancer cells. It is expected that drugs will accumulate inside cells and kill cancer cells (Non-Patent Documents 1-5) .

[0003] As one of the antibody-drug conjugates, we have developed a conjugate that combines an antibody and a topoisomerase I inhibitor called exon Antibody-drug conjugates that contain satecan as a component are known (Patent Documents 1 to 8, Non-patent literature 6, 7). These antibody-drug conjugates have excellent antitumor effects and safety. As this compound has potential applications, clinical trials are currently underway.

[0004] Antibody-drug conjugate formulations include antibody-drug conjugates that contain maytansinoids as components. Drug conjugate preparations (Patent Documents 9 to 12), containing monomethyl auristatin E as a component and antibody-drug conjugate formulations comprising SN-38 (Patent Documents 13 to 15). Preparations of antibody-drug conjugates that contain the compound as a component (Patent Document 16) are known. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] International Publication No. 2014 / 057687 [Patent Document 2] International Publication No. 2014 / 061277 [Patent Document 3] International Publication No. 2015 / 098099 [Patent Document 4] International Publication No. 2015 / 115091 [Patent Document 5] International Publication No. 2015 / 146132 [Patent Document 6] International Publication No. 2015 / 155976 [Patent Document 7] International Publication No. 2015 / 155998 [Patent Document 8] International Publication No. 2018 / 135501 [Patent Document 9] International Publication No. 2004 / 004639 [Patent Document 10] International Publication No. 2004 / 110498 [Patent Document 11] International Publication No. 2007 / 019232 [Patent Document 12] International Publication No. 2015 / 059147 [Patent Document 13] International Publication No. 2010 / 081004 [Patent Document 14] International Publication No. 2014 / 143765 [Patent Document 15] International Publication No. 2015 / 157286 [Patent Document 16] International Publication No. 2014 / 092804 [Non-patent literature]

[0006] [Non-Patent Document 1] Ducry, L., et al., Bioconjugate Chem. (2010) 21, 5-13. [Non-Patent Document 2] Alley, SC, et al., Current Opinion in Chemical Biology (2010) 14, 529-537. [Non-Patent Document 3] Damle NK Expert Opin. Biol. Ther. (2004) 4, 1445-1452. [Non-Patent Document 4] Senter PD, et al., Nature Biotechnology (2012) 30, 631-637. [Non-Patent Document 5] Howard A. et al., J Clin Oncol 29: 398-405. [Non-Patent Document 6] Ogitani Y. et al., Clinical Cancer Research (2016) 22(20), 5097-5108. [Non-Patent Document 7] Ogitani Y. et al., Cancer Science (2016) 107, 1039-1046. Summary of the Invention [Problem to be solved by the invention]

[0007] The formation of aggregates and degradation products in antibody formulations may be a cause of immune dysfunction in patients receiving the formulation. It can have undesirable medical effects, such as causing hypersensitivity and venous disorders. In the formulation of drugs, it is necessary to suppress the formation of aggregates and degradation products. Therefore, various pharmaceutical compositions (e.g., aqueous injections and freeze-dried injections) are being investigated. However, when formulating an antibody-drug conjugate, it is necessary to consider not only the antibody portion but also the drug linker. The specific physical properties of the anchor part must also be taken into account, necessitating more complex considerations. In addition, when preparing a freeze-dried injection from an aqueous solution containing sucrose or trehalose, (1) The primary drying process takes a long time, and (2) the freeze-dried cake shrinks. There are problems such as the following: The present invention prevents the formation of aggregates and degradation products in specific antibody-drug conjugates. Inhibited pharmaceutical compositions (especially aqueous injections and lyophilized injections), and lyophilized from aqueous solutions. Providing an efficient freeze-drying method for injections is a major challenge. [Means for solving the problem]

[0008] The present inventors have demonstrated that the formation of aggregates and degradation products in a specific antibody-drug conjugate The present inventors have discovered pharmaceutical compositions (particularly aqueous injections and freeze-dried injections) that suppress the formation of benzodiazepines. An efficient freeze-drying method was discovered for turning a solution into a freeze-dried injectable formulation.

[0009] That is, the present invention provides [1] (i) antibody-drug conjugates, (ii) a histidine buffer; (iii) sucrose or trehalose, and (iv) surfactants, a pharmaceutical composition comprising: (wherein the antibody-drug conjugate has the following formula:

[0010] [ka]

[0011] (wherein A represents the binding site to the antibody) The antibody-drug conjugate is formed by binding the drug linker represented by the formula: (It is a jugate). [2] The pharmaceutical composition according to [1], wherein the surfactant is polysorbate 80 or polysorbate 20. composition. [3] (i) per 20 mg of antibody-drug conjugate, (ii) 3 to 80 mmol of a histidine buffer; (iii) 24 to 320 mg of sucrose or trehalose, and (iv) 0.05 to 1.6 mg of polysorbate 80 or polysorbate 20, The pharmaceutical composition according to [1] or [2], [4] (i) per 20 mg of antibody-drug conjugate, (ii) 10 to 40 mmol of histidine buffer; (iii) 90 mg of sucrose or 100 mg of trehalose hydrate, and (iv) 0.2 to 0.4 mg of polysorbate 80 or polysorbate 20, The pharmaceutical composition according to any one of [1] to [3], [5] (i) antibody-drug conjugates, (ii) a histidine buffer; (iii) sucrose, and (iv) Polysorbate 80 or Polysorbate 20 The pharmaceutical composition according to [1] or [2],

[0012] [6] (i) per 20 mg of antibody-drug conjugate, (ii) 3 to 80 mmol of a histidine buffer; (iii) 24 to 320 mg of sucrose, and (iv) 0.05 to 1.6 mg of polysorbate 80 or polysorbate 20, The pharmaceutical composition according to [1], [2] or [5], [7] (i) per 20 mg of antibody-drug conjugate, (ii) 10 to 40 mmol of histidine buffer; (iii) 90 mg of sucrose, and (iv) 0.2 to 0.4 mg of polysorbate 80 or polysorbate 20, The pharmaceutical composition according to [1], [2], [5] or [6], [8] (i) per 20 mg of antibody-drug conjugate, (ii) 10 or 25 mmol of histidine buffer; (iii) 90 mg of sucrose, and (iv) 0.2 or 0.3 mg of polysorbate 80 or polysorbate 20, The pharmaceutical composition according to any one of [1] to [7], [9] The pH of the antibody-drug conjugate dissolved in water at a concentration of 20 mg / mL is The pharmaceutical composition according to any one of [1] to [8], wherein the viscosity is 4.0 to 7.0.

[10] The average number of drug linkers per antibody in the antibody-drug conjugate ranges from 2 to The pharmaceutical composition according to any one of claims 1 to 9, wherein the number of active ingredients is in the range of 8.

[0013]

[11] The antibody in the antibody-drug conjugate is an anti-HER2 antibody, an anti-HER3 antibody, an anti-TR antibody, Any of [1] to

[10] , which is an OP2 antibody, an anti-B7-H3 antibody, or an anti-GPR20 antibody. The pharmaceutical composition according to any one of claims 1 to 5.

[12]

[11] The antibody in the antibody-drug conjugate is an anti-HER2 antibody. Pharmaceutical compositions.

[13] The anti-HER2 antibody comprises an amino acid sequence set forth in amino acid numbers 1 to 449 in SEQ ID NO:1. and a heavy chain consisting of the amino acid sequence set forth in amino acid numbers 1 to 214 in SEQ ID NO:2. or a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO: 1 and and a light chain having the amino acid sequence set forth in SEQ ID NO:2. The pharmaceutical composition described above.

[14] The average number of drug linkers per antibody in antibody-drug conjugates ranged from 7 to The pharmaceutical composition according to

[12] or

[13] , wherein the number of active ingredients is in the range of 8.

[15] (i) per 20 mg of antibody-drug conjugate, (ii) 25 mmol of histidine buffer; (iii) 90 mg of sucrose, and (iv) 0.3 mg of polysorbate 80, wherein the antibody-drug conjugate is dissolved in water at a concentration of 20 mg / mL. The pharmaceutical composition according to any one of

[12] to

[14] , wherein the pH is 5.5.

[0014]

[16] (i) per 20 mg of antibody-drug conjugate, (ii) 0.89 mg L-histidine and 4.04 mg L-histidine hydrochloride in water Japanese food, (iii) 90 mg of sucrose; and (iv) any one of

[12] to

[14] containing 0.3 mg of polysorbate 80 Item 1. The pharmaceutical composition according to item 1.

[17] (i) 100 mg of the antibody-drug conjugate; (ii) 4.45 mg of L-histidine and 20.2 mg of L-histidine hydrochloride in water Japanese food, (iii) 450 mg of sucrose; and (iv) any one of

[12] to

[14] containing 1.5 mg of polysorbate 80 Item 1. The pharmaceutical composition according to item 1.

[18]

[11] The antibody in the antibody-drug conjugate is an anti-HER3 antibody. Pharmaceutical compositions.

[19] The anti-HER3 antibody comprises a heavy chain having the amino acid sequence shown in SEQ ID NO: 3 and a heavy chain having the amino acid sequence shown in SEQ ID NO: 4. or the carboxyl terminus of the heavy chain of said antibody, The pharmaceutical composition according to

[18] , wherein the antibody is an antibody in which the lysine residues are deleted.

[20] The average number of drug linkers per antibody in antibody-drug conjugates ranged from 7 to The pharmaceutical composition according to

[18] or

[19] , wherein the number of active ingredients is in the range of 8.

[0015] [twenty one] (i) per 20 mg of antibody-drug conjugate, (ii) 25 mmol of histidine buffer; (iii) 90 mg of sucrose, and (iv) 0.3 mg of polysorbate 20, wherein the antibody-drug conjugate is dissolved in water at a concentration of 20 mg / mL. The pharmaceutical composition according to any one of

[18] to

[20] , wherein the pH is 5.4. [twenty two] (i) per 20 mg of antibody-drug conjugate, (ii) 0.81 mg of L-histidine and 4.14 mg of L-histidine hydrochloride in water Japanese food, (iii) 90 mg of sucrose; and (iv) Any one of

[18] to

[20] containing 0.3 mg of polysorbate 20. Item 1. The pharmaceutical composition according to item 1. [twenty three] (i) 100 mg of the antibody-drug conjugate; (ii) 4.06 mg of L-histidine, and 20.7 mg of L-histidine hydrochloride in water Japanese food, (iii) 450 mg of sucrose; and (iv) any one of

[18] to

[20] containing 1.5 mg of polysorbate 20 Item 1. The pharmaceutical composition according to item 1. [twenty four]

[11] , wherein the antibody in the antibody-drug conjugate is an anti-TROP2 antibody. A pharmaceutical composition comprising: [twenty five] The anti-TROP2 antibody comprises the amino acid sequence set forth in amino acid numbers 20 to 470 of SEQ ID NO:5. and a heavy chain consisting of the amino acid sequence set forth in amino acid numbers 21 to 234 in SEQ ID NO:6. or a light chain of said antibody comprising a lysine residue at the carboxyl terminus of said heavy chain. The pharmaceutical composition according to

[24] , wherein the antibody is an antibody having a deletion of the group.

[0016]

[26] The average number of drug linkers per antibody in antibody-drug conjugates ranges from 3 to The pharmaceutical composition according to

[24] or

[25] , wherein the range is 5.

[27] (i) per 20 mg of antibody-drug conjugate, (ii) 10 mmol of histidine buffer; (iii) 90 mg of sucrose, and (iv) 0.2 or 0.3 mg of polysorbate 80, wherein the antibody-drug conjugate is dissolved in water at a concentration of 20 mg / mL. The pharmaceutical composition according to any one of

[24] to

[26] , wherein the pH of the pharmaceutical composition is 6.0.

[28] (i) per 20 mg of antibody-drug conjugate, (ii) 0.78 mg L-histidine and 1.05 mg L-histidine hydrochloride in water Japanese food, (iii) 90 mg of sucrose; and (iv)

[24] to

[26] , containing 0.2 or 0.3 mg of polysorbate 80 The pharmaceutical composition according to any one of claims 1 to 4.

[29] (i) 100 mg of the antibody-drug conjugate; (ii) 3.88 mg of L-histidine and 5.26 mg of L-histidine hydrochloride in water Japanese food, (iii) 450 mg of sucrose; and (iv)

[24] to

[26] , each of which contains 1.0 or 1.5 mg of polysorbate 80. The pharmaceutical composition according to any one of claims 1 to 4.

[30]

[11] , wherein the antibody in the antibody-drug conjugate is an anti-B7-H3 antibody. A pharmaceutical composition comprising:

[0017]

[31] 7. The anti-B7-H3 antibody comprises the amino acid sequence set forth in amino acid numbers 20 to 471 of SEQ ID NO:7. and a heavy chain consisting of the amino acid sequence set forth in amino acid numbers 21 to 233 in SEQ ID NO:8. or a light chain of said antibody comprising a lysine residue at the carboxyl terminus of said heavy chain. The pharmaceutical composition according to

[30] , wherein the antibody is an antibody having a deletion of the group.

[32] The average number of drug linkers per antibody in antibody-drug conjugates ranges from 3 to The pharmaceutical composition according to

[30] or

[31] , wherein the range is 5.

[33] (i) per 20 mg of antibody-drug conjugate, (ii) 10 mmol of histidine buffer; (iii) 90 mg of sucrose, and (iv) 0.2 or 0.3 mg of polysorbate 20, wherein the antibody-drug conjugate is dissolved in water at a concentration of 20 mg / mL. The pharmaceutical composition according to any one of

[30] to

[32] , wherein the pH is 5.9.

[34] (i) per 20 mg of antibody-drug conjugate, (ii) 0.65 mg L-histidine and 1.22 mg L-histidine hydrochloride in water Japanese food, (iii) 90 mg of sucrose; and (iv)

[30] to

[32] , containing 0.2 or 0.3 mg of polysorbate 20 The pharmaceutical composition according to any one of claims 1 to 4.

[35] (i) 100 mg of the antibody-drug conjugate; (ii) 3.23 mg of L-histidine, and 6.12 mg of L-histidine hydrochloride in water Japanese food, (iii) 450 mg of sucrose; and (iv)

[30] to

[32] , containing 1.0 or 1.5 mg of polysorbate 20. The pharmaceutical composition according to any one of claims 1 to 4.

[0018]

[36]

[11] , wherein the antibody in the antibody-drug conjugate is an anti-GPR20 antibody. A pharmaceutical composition comprising:

[37] The anti-GPR20 antibody is selected from the group consisting of amino acids 20 to 472 of SEQ ID NO:9. and a heavy chain consisting of the amino acid sequence set forth in amino acid numbers 21 to 234 in SEQ ID NO: 10. or a light chain of said antibody comprising a carboxyl-terminal lysine of said heavy chain. The pharmaceutical composition according to

[36] , wherein the antibody has deleted residues.

[38] The average number of drug linkers per antibody in antibody-drug conjugates ranged from 7 to The pharmaceutical composition according to

[36] or

[37] , wherein the number of active ingredients is in the range of 8.

[39] (i) per 20 mg of antibody-drug conjugate, (ii) 10 mmol of histidine buffer; (iii) 90 mg of sucrose, and (iv) 0.3 mg of polysorbate 80, wherein the antibody-drug conjugate is dissolved in water at a concentration of 20 mg / mL. The pharmaceutical composition according to any one of

[36] to

[38] , wherein the pH is 5.4.

[40] (i) per 20 mg of antibody-drug conjugate, (ii) 0.32 mg of L-histidine and 1.66 mg of L-histidine hydrochloride in water Japanese food, (iii) 90 mg of sucrose; and (iv) any one of

[36] to

[38] containing 0.3 mg of polysorbate 80 Item 1. The pharmaceutical composition according to item 1.

[0019]

[41] (i) 100 mg of the antibody-drug conjugate; (ii) 1.62 mg of L-histidine, and 8.29 mg of L-histidine hydrochloride in water Japanese food, (iii) 450 mg of sucrose; and (iv) any one of

[36] to

[38] containing 1.5 mg of polysorbate 80 Item 1. The pharmaceutical composition according to item 1.

[42] (i) per 20 mg of antibody-drug conjugate, (ii) 0.89 mg L-histidine and 4.04 mg L-histidine hydrochloride in water Japanese food, (iii) 90 mg of sucrose; and (iv) a pharmaceutical composition comprising 0.3 mg of polysorbate 80; (wherein the antibody-drug conjugate has the following formula:

[0020] [ka]

[0021] (wherein A represents the binding site to the antibody) The antibody-drug conjugate is formed by binding the drug linker represented by the formula: (It is a jugate).

[43] (i) 100 mg of the antibody-drug conjugate; (ii) 4.45 mg of L-histidine and 20.2 mg of L-histidine hydrochloride in water Japanese food, (iii) 450 mg of sucrose; and (iv) The pharmaceutical composition described in

[42] , containing 1.5 mg of polysorbate 80.

[44] The average number of drug linkers per antibody in antibody-drug conjugates ranged from 7 to The pharmaceutical composition according to

[42] or

[43] , wherein the number of active ingredients is in the range of 8.

[45] The pharmaceutical composition described in any one of

[42] to

[0044] , wherein the antibody in the antibody-drug conjugate is an anti-HER2 antibody.

[0022]

[46] The anti-HER2 antibody comprises an amino acid sequence set forth in amino acid numbers 1 to 449 in SEQ ID NO:1. and a heavy chain consisting of the amino acid sequence set forth in amino acid numbers 1 to 214 in SEQ ID NO:2. The pharmaceutical composition according to

[45] , wherein the antibody comprises a light chain consisting of:

[47] The anti-HER2 antibody comprises a heavy chain having the amino acid sequence shown in SEQ ID NO: 1 and a heavy chain having the amino acid sequence shown in SEQ ID NO: 2. The pharmaceutical composition according to

[45] , wherein the antibody comprises a light chain having the amino acid sequence set forth above.

[48] (i) per 20 mg of antibody-drug conjugate, (ii) 0.81 mg of L-histidine and 4.14 mg of L-histidine hydrochloride in water Japanese food, (iii) 90 mg of sucrose; and (iv) a pharmaceutical composition comprising 0.3 mg of polysorbate 20; (wherein the antibody-drug conjugate has the following formula:

[0023] [ka]

[0024] (wherein A represents the binding site to the antibody) The antibody-drug conjugate is formed by binding the drug linker represented by the formula: (It is a jugate).

[49] (i) 100 mg of the antibody-drug conjugate; (ii) 4.06 mg of L-histidine, and 20.7 mg of L-histidine hydrochloride in water Japanese food, (iii) 450 mg of sucrose; and (iv) The pharmaceutical composition described in

[48] , which contains 1.5 mg of polysorbate 20.

[50] The average number of drug linkers per antibody in antibody-drug conjugates ranged from 7 to The pharmaceutical composition according to

[48] or

[49] , wherein the number of active ingredients is in the range of 8.

[0025]

[51] The pharmaceutical composition described in any one of

[48] to

[0050] , wherein the antibody in the antibody-drug conjugate is an anti-HER3 antibody.

[52] The anti-HER3 antibody comprises a heavy chain having the amino acid sequence shown in SEQ ID NO: 3 and a heavy chain having the amino acid sequence shown in SEQ ID NO: 4. The pharmaceutical composition according to

[51] , wherein the antibody comprises a light chain having the amino acid sequence described above.

[53] The anti-HER3 antibody lacks a lysine residue at the carboxyl terminus of the heavy chain, as described in

[52] . A pharmaceutical composition comprising:

[54] (i) per 20 mg of antibody-drug conjugate, (ii) 0.78 mg L-histidine and 1.05 mg L-histidine hydrochloride in water Japanese food, (iii) 90 mg of sucrose; and (iv) a pharmaceutical composition comprising 0.2 or 0.3 mg of polysorbate 80; (wherein the antibody-drug conjugate has the following formula:

[0026] [ka]

[0027] (wherein A represents the binding site to the antibody) The antibody-drug conjugate is formed by binding the drug linker represented by the formula: (It is a jugate).

[55] (i) 100 mg of the antibody-drug conjugate; (ii) 3.88 mg of L-histidine and 5.26 mg of L-histidine hydrochloride in water Japanese food, (iii) 450 mg of sucrose; and (iv) The pharmaceutical composition according to

[54] , which contains 1.0 or 1.5 mg of polysorbate 80. Composition.

[56] The average number of drug linkers per antibody in antibody-drug conjugates ranges from 3 to The pharmaceutical composition according to

[54] or

[55] , wherein the number of active ingredients is in the range of 5.

[57] The pharmaceutical composition according to any one of

[54] to

[0056] , wherein the antibody in the antibody-drug conjugate is an anti-TROP2 antibody.

[58] The anti-TROP2 antibody comprises the amino acid sequence set forth in amino acid numbers 20 to 470 of SEQ ID NO:5. and a heavy chain consisting of the amino acid sequence set forth in amino acid numbers 21 to 234 in SEQ ID NO:6. The pharmaceutical composition described in

[57] , wherein the antibody comprises a light chain consisting of an amino acid sequence.

[59] The anti-TROP2 antibody lacks a lysine residue at the carboxyl terminus of the heavy chain, as described in

[58] . The pharmaceutical composition described above.

[60] (i) per 20 mg of antibody-drug conjugate, (ii) 0.65 mg L-histidine and 1.22 mg L-histidine hydrochloride in water Japanese food, (iii) 90 mg of sucrose; and (iv) a pharmaceutical composition comprising 0.2 or 0.3 mg of polysorbate 20; (wherein the antibody-drug conjugate has the following formula:

[0028] [ka]

[0029] (wherein A represents the binding site to the antibody) The antibody-drug conjugate is formed by binding the drug linker represented by the formula: (It is a jugate).

[61] (i) 100 mg of the antibody-drug conjugate; (ii) 3.23 mg of L-histidine, and 6.12 mg of L-histidine hydrochloride in water Japanese food, (iii) 450 mg of sucrose; and (iv) The pharmaceutical composition according to

[60] , which contains 1.0 or 1.5 mg of polysorbate 20. Composition.

[62] The average number of drug linkers per antibody in antibody-drug conjugates ranges from 3 to The pharmaceutical composition according to

[60] or

[61] , wherein the number of active ingredients is in the range of 5.

[63] The pharmaceutical composition described in any one of

[60] to

[0062] , wherein the antibody in the antibody-drug conjugate is an anti-B7-H3 antibody.

[64] 7. The anti-B7-H3 antibody comprises the amino acid sequence set forth in amino acid numbers 20 to 471 of SEQ ID NO:7. and a heavy chain consisting of the amino acid sequence set forth in amino acid numbers 21 to 233 in SEQ ID NO:8. The pharmaceutical composition according to

[63] , wherein the antibody comprises a light chain consisting of an amino acid sequence.

[65] The anti-B7-H3 antibody lacks a lysine residue at the carboxyl terminus of the heavy chain, as described in

[64] . The pharmaceutical composition described above.

[66] (i) per 20 mg of antibody-drug conjugate, (ii) 0.32 mg of L-histidine and 1.66 mg of L-histidine hydrochloride in water Japanese food, (iii) 90 mg of sucrose; and (iv) a pharmaceutical composition comprising 0.3 mg of polysorbate 80; (wherein the antibody-drug conjugate has the following formula:

[0030] [ka]

[0031] (wherein A represents the binding site to the antibody) The antibody-drug conjugate is formed by binding the drug linker represented by the formula: (It is a jugate).

[67] (i) 100 mg of the antibody-drug conjugate; (ii) 1.62 mg of L-histidine, and 8.29 mg of L-histidine hydrochloride in water Japanese food, (iii) 450 mg of sucrose; and (iv) The pharmaceutical composition described in

[66] , containing 1.5 mg of polysorbate 80.

[68] The average number of drug linkers per antibody in antibody-drug conjugates ranged from 7 to The pharmaceutical composition according to

[66] or

[67] , wherein the number of active ingredients is in the range of 8.

[69] The pharmaceutical composition described in any one of

[66] to

[0068] , wherein the antibody in the antibody-drug conjugate is an anti-GPR20 antibody.

[70] The anti-GPR20 antibody is selected from the group consisting of amino acids 20 to 472 of SEQ ID NO:9. and a heavy chain consisting of the amino acid sequence set forth in amino acid numbers 21 to 234 in SEQ ID NO: 10. The pharmaceutical composition according to

[69] , wherein the antibody comprises a light chain consisting of an amino acid sequence.

[0032]

[71] The anti-GPR20 antibody has a deletion of the lysine residue at the carboxyl terminus of the heavy chain, as described in

[70] . The pharmaceutical composition described above.

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

[71] , which is an injectable preparation.

[73] The pharmaceutical composition described in

[72] , which is an aqueous injectable formulation.

[74] The pharmaceutical composition according to

[73] , wherein the concentration of the antibody-drug conjugate is 20 mg / mL. thing.

[75] The pharmaceutical composition according to

[73] or

[74] , which is frozen.

[0033]

[76] The pharmaceutical composition described in

[72] , which is a lyophilized injection preparation.

[77] The pharmaceutical composition described in

[76] , contained in an amber vial.

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

[77] , which is for the treatment of cancer.

[79] (1) A specified amount of (i) antibody-drug conjugates, (ii) a histidine buffer; (iii) sucrose or trehalose, and (iv) surfactants, preparing an aqueous solution comprising (2) if necessary, adjusting the pH of the aqueous solution to a predetermined value, and then (3) freeze-drying the aqueous solution; A method for producing the pharmaceutical composition described in

[76] , comprising:

[80] The process of freeze-drying the aqueous solution is a process of annealing at a shelf temperature near the eutectic point of the aqueous solution.

[79] , comprising the process for producing the same; (Here, near the eutectic point means a temperature 1.5°C lower than the eutectic point to a temperature 1.5°C higher than the eutectic point. (Indicates high temperature range).

[0034]

[81] The process of freeze-drying the aqueous solution is a process of annealing at a shelf temperature equal to the eutectic point of the aqueous solution. The method for producing the same according to

[80] ,

[82] Compared with the case where annealing was performed at a shelf temperature 5°C lower than the eutectic point, the primary drying process The method according to

[80] or

[81] , wherein the process time is shortened.

[83] Compared to annealing at a shelf temperature 5°C lower than the eutectic point, there was less shrinkage. Any one of

[80] to

[82] , characterized in that a small amount of freeze-dried cake is obtained. The manufacturing method described in item .

[84] The process of freeze-drying the aqueous solution includes an annealing process at a shelf temperature of -4 to -1°C. The manufacturing method described in

[79] .

[85] The process of freeze-drying the aqueous solution includes an annealing process at a shelf temperature of -4 to -2°C. The manufacturing method described in

[79] .

[0035]

[86] The step of freeze-drying the aqueous solution includes a process of annealing at a shelf temperature of -2.5°C. The manufacturing method described in

[79] .

[87] The process of freeze-drying the aqueous solution is carried out by first drying at a shelf temperature of -5 to 5°C and under a vacuum of 5 to 15 Pa. The method according to any one of

[84] to

[86] , further comprising a drying process.

[88] The process of freeze-drying the aqueous solution is a process in which the first drying is performed at a shelf temperature of 0°C and under a vacuum of 10 Pa. The method according to any one of

[84] to

[86] , further comprising the step of:

[89] The process of freeze-drying the aqueous solution is carried out at a shelf temperature of 40 to 50°C and under a vacuum of 5 to 15 Pa. The method according to

[87] or

[88] , further comprising a drying process.

[90] The process of freeze-drying the aqueous solution is a process of secondary drying at a shelf temperature of 45°C and under a vacuum of 10 Pa. The method according to

[87] or

[88] , further comprising a process for producing the same.

[0036]

[91] A kit comprising the pharmaceutical composition of

[42] or

[43] and water for injection.

[92] The kit according to

[91] , wherein the water for injection is stored in an ampoule.

[93] Annealing of aqueous solutions containing sucrose or trehalose at shelf temperatures near the eutectic point (wherein "near the eutectic point" means a point lower than the eutectic point) (The range of temperatures is from 1.5°C lower to 1.5°C higher than the eutectic point.)

[94] Annealing at a shelf temperature equal to the eutectic point of the aqueous solution containing sucrose or trehalose A method for producing a freeze-dried injection, comprising the steps of: Regarding. Effect of the Invention

[0037] According to the present invention, the formation of aggregates and degradation products in a specific antibody-drug conjugate can be prevented. A pharmaceutical composition (particularly an aqueous injection and a freeze-dried injection) in which the formation of benzodiazepines is suppressed, and It is possible to provide an efficient freeze-drying method for freeze-dried injections. [Brief description of the drawings]

[0038] [Figure 1] FIG. 1 shows the amino acid sequence of the anti-HER2 antibody heavy chain (SEQ ID NO:1). [Diagram 2] FIG. 2 shows the amino acid sequence of the anti-HER2 antibody light chain (SEQ ID NO:2). [Diagram 3] This is a diagram related to excipient screening. The horizontal axis shows the time course for each formulation, and the vertical axis shows the content rate of aggregates. [Figure 4] This is a diagram relating to buffer screening. The horizontal axis shows time progression for each formulation, and the vertical axis shows the aggregate content. [Diagram 5] This is a diagram related to pH screening. The horizontal axis shows the time progression for each formulation, and the vertical axis shows the content of aggregates. [Figure 6] This is a diagram related to pH screening. The horizontal axis shows the time progression for each formulation, and the vertical axis shows the NPI content. [Figure 7] This is a diagram related to surfactant screening. The horizontal axis shows the time progression for each formulation, and the vertical axis shows the content of insoluble particles smaller than 10 mm. Here, "PS" means polysorbate, "PS20" means polysorbate 20, and "PS80" means polysorbate 80. [Figure 8] 1 is a diagram relating to vial studies, in which the horizontal axis indicates the amount of light irradiation and the vertical axis indicates the content of aggregates. [Figure 9] This is a diagram related to vial studies. The horizontal axis shows the amount of light irradiation, and the vertical axis shows the IoP content. [Figure 10]This is a contour plot of the product temperature of the aqueous solution (1) when the shelf temperature and the degree of vacuum inside the chamber are varied in the primary drying process.

[0039] [Figure 11] This is a contour plot of the drying time of the aqueous solution (1) when the shelf temperature and the degree of vacuum inside the chamber are changed in the primary drying process. [Figure 12] FIG. 1 shows the product temperature (during annealing) and the shape of the freeze-dried cake when annealing was performed at shelf temperatures ranging from −7° C. to 0.5° C. and then freeze-drying. [Figure 13] FIG. 1 shows the thermal denaturation of antibody-drug conjugate (1) by capillary differential scanning calorimetry. [Figure 14] FIG. 2 shows the amino acid sequence of the anti-HER3 antibody heavy chain (SEQ ID NO: 3). [Figure 15] FIG. 1 shows the amino acid sequence of the anti-HER3 antibody light chain (SEQ ID NO: 4). [Figure 16] FIG. 1 shows the amino acid sequence of the anti-TROP2 antibody heavy chain (SEQ ID NO:5). [Figure 17] FIG. 1 shows the amino acid sequence of the anti-TROP2 antibody light chain (SEQ ID NO: 6). [Figure 18] FIG. 7 shows the amino acid sequence of the anti-B7-H3 antibody heavy chain (SEQ ID NO:7). [Figure 19] FIG. 1 shows the amino acid sequence of the anti-B7-H3 antibody light chain (SEQ ID NO: 8). [Figure 20] FIG. 1 shows the amino acid sequence of the anti-GPR20 antibody heavy chain (SEQ ID NO: 9). [Figure 21] FIG. 1 shows the amino acid sequence of the anti-GPR20 antibody light chain (SEQ ID NO: 10). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0040] Preferred embodiments of the present invention will now be described with reference to the drawings. The embodiment described below is an example of a typical embodiment of the present invention. This will not narrow the scope of the present invention.

[0041] [Antibody-drug conjugates] The antibody-drug conjugate used in the present invention has the formula:

[0042] [ka]

[0043] (wherein A represents the binding site to the antibody) The antibody-drug complex is formed by binding the drug linker represented by the formula: It is a njugate.

[0044] In the present invention, the portion of the antibody-drug conjugate consisting of a linker and a drug The structure is called a "drug linker." This drug linker is a drug that binds to the disulfide bond between the antibody chains. Thiol groups (in other words) generated at the positions (two heavy chain-heavy chain positions and two heavy chain-light chain positions) in other words, it is bound to the sulfur atom of a cysteine ​​residue.

[0045] The drug linker of the present invention is a topoisomerase I inhibitor, exatecan (IUPAC Name: (1S,9S)-1-amino-9-ethyl-5-fluoro-1,2,3,9,12, 15-Hexahydro-9-hydroxy-4-methyl-10H,13H-benzo[de]pi Lano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione Chemical name: (1S,9S)-1-amino-9-ethyl-5-fluoro-2,3-dihydro 9-Hydroxy-4-methyl-1H,12H-benzo[de]pyrano[3',4': 6,7]indolizino[1,2-b]quinoline-10,13(9H,15H)-dione and Exatecan is a compound of the formula

[0046] [ka]

[0047] It is a camptothecin derivative having an antitumor effect, represented by the following formula:

[0048] The antibody-drug conjugate used in the present invention may also be represented by the following formula:

[0049] [ka]

[0050] Here, the drug linker is bound to the antibody via a thioether bond. The so-called average drug binding ratio (DAR; Drug-to-Antibody Ratio) and It is synonymous with and indicates the average number of drug linkers bound per antibody. The antibody-drug conjugate used in the present invention is capable of transporting the antibody into cancer cells and reacting with the antibody according to the following formula:

[0051] [ka]

[0052] A compound having a topoisomerase I inhibitory activity represented by the formula (hereinafter referred to as "compound (1)"). It exerts an antitumor effect by releasing agonists. Compound (1) is a compound that is capable of reacting with the antibody-drug conjugate of the present invention after the linker moiety is cleaved. The following formula is thought to be caused by

[0053] [ka]

[0054] This is believed to occur due to decomposition of the aminal structure of the compound represented by the following formula:

[0055] The antibody-drug conjugates used in the present invention may also have a bystander effect. It is known (Ogitani Y. et al., Cancer Science (2016) 107, 1039-1046). This bystander effect is believed to be due to the fact that the antibody-drug conjugates used in the present invention inhibit the expression of the target After internalization into cancer cells, compound (1) is released and targets nearby cells that do not express the target. It also exerts an antitumor effect on cancer cells.

[0056] [Antibodies in antibody-drug conjugates] The antibody in the antibody-drug conjugate used in the present invention may be derived from any species. The antibodies may be derived from any source, but are preferably derived from humans, rats, mice, and rabbits. If derived from a species other than human, it is preferred to chimerize or humanize it using well-known techniques. The antibody of the present invention may be a polyclonal antibody or a monoclonal antibody. Although antibodies may be used, monoclonal antibodies are preferred.

[0057] The antibody in the antibody-drug conjugate used in the present invention preferably targets cancer cells. It has the property of being able to recognize cancer cells and bind to them. the ability to be taken up and internalized into cancer cells, and / or the ability to kill cancer cells It is preferable that the material has properties such as

[0058] The binding of the antibody to the cancer cells can be confirmed using flow cytometry. Antibody uptake into the cell can be achieved by (1) intracellularly delivering a secondary antibody (fluorescently labeled) that binds to the therapeutic antibody. A Cell Death and Differentiation Assay (CDA) to visualize the antibodies incorporated into the cells using a fluorescent microscope. n (2008) 15, 751-761), (2) using a secondary antibody (fluorescently labeled) that binds to the therapeutic antibody to identify cells. An assay to measure the amount of fluorescence incorporated into the 5268-5282, December 2004), or (3) using an immunotoxin that binds to the therapeutic antibody, When taken up into cells, Mab-ZAP releases toxins that inhibit cell proliferation. This can be confirmed using the immunotoxicity assay (BioTechniques 28:162-165, January 2000). The recombinant complex protein of the catalytic domain of diphtheria toxin and protein G is can also be used.

[0059] Anti-tumor activity of antibodies can be evaluated in vitro by measuring their ability to inhibit cell proliferation. For example, a cancer cell line that overexpresses the antibody's target protein can be cultured and Antibodies were added at various concentrations to the cells, and the effects on focus formation, colony formation, and spheroid growth were evaluated. In vivo, for example, the inhibitory activity against a target protein can be measured. The antibody is administered to nude mice transplanted with a cancer cell line expressing the antibody, and changes in the cancer cells are measured. By this, antitumor activity can be confirmed.

[0060] It is preferable that the antibody itself has an antitumor effect, but the antibody-drug conjugate may have an antitumor effect. Since a compound that exerts an antitumor effect is bound to the antibody, the antibody itself does not necessarily have an antitumor effect. For the purpose of specifically and selectively exerting the cytotoxicity of antitumor compounds on cancer cells, It is important, and preferred, that the antibody has the property of being internalized and transported into cancer cells. The antibody in the antibody-drug conjugate used in the present invention can be obtained by known means. For example, antigenic polypeptides can be isolated by methods commonly used in the art. The antibody is obtained by immunizing an animal with a peptide and collecting and purifying the antibodies produced in the body. The origin of the antigen is not limited to humans, but may be from animals other than humans, such as mice and rats. Alternatively, animals can be immunized with a heterologous antigen. In this case, the obtained antibodies that bind to the heterologous antigen are By testing the cross-reactivity between the antibody and human antigens, it is possible to select antibodies that can be applied to human diseases. Cut.

[0061] Also, known methods (e.g., Kohler and Milstein, Nature (1975) 256, p. 495-497; Ke nnet, R. ed., Monoclonal Antibodies, p.365-367, Plenum Press, NY (1980)). By fusing antibody-producing cells that produce antibodies against antigens with myeloma cells, It is also possible to establish hybridomas and obtain monoclonal antibodies.

[0062] The antigen is produced by genetically engineering the gene encoding the antigen protein in the host cell. Specifically, a vector capable of expressing an antigen gene is prepared. This can then be introduced into a host cell to express the gene, and the expressed antigen can be purified. Immunizing an animal with the antigen-expressing cells or cell lines expressing the antigen by the above genetic manipulation. The antibodies can also be obtained by using the method.

[0063] The antibody in the antibody-drug conjugate used in the present invention is a heterologous antigen for humans. Recombinant antibodies that have been artificially modified to reduce their affinity, for example, Preferably, the antibody is a chimeric antibody or a humanized antibody. Alternatively, it is preferable that the antibody is an antibody having only the gene sequence of an antibody derived from a human, i.e., a human antibody. These antibodies can be produced using known methods.

[0064] Chimeric antibodies include antibodies in which the variable and constant regions of the antibody are of different species, such as mouse antibodies. Chimeric antibodies in which the variable regions of a mouse or rat-derived antibody are joined to constant regions of human origin are also included. (Proc. Natl. Acad. Sci. USA, 81, 6851-6855, (1984)).

[0065] Humanized antibodies are characterized by the complementarity determining regions (CDRs) of heterologous antibodies. The antibody was developed by incorporating only the ity determining region (IL-1) of human-derived antibodies. (Nature (1986) 321, p.522-525), and the CDR grafting method is used to graft the CDR sequences of heterologous antibodies. In addition, some of the amino acid residues in the framework of the xenoantibody have been transplanted into a human antibody ( International Publication No. 90 / 07861, gene conversion mutagenesis Antibodies humanized using a cytogenetic mutagenesis (US Pat. No. 5,633,363) strategy (US Pat. No. 5,633,363). No. 21337).

[0066] Human antibodies include those derived from human antibodies having a human chromosome fragment containing the genes for the heavy and light chains of human antibodies. Antibodies generated using antibody-producing mice (Tomizuka, K. et al., Nature Genetics (1997) 1 6, p.133-143;Kuroiwa, Y. et. al., Nucl. Acids Res.(1998) 26, p.3447-3448;Yoshida , H. et. al., Animal Cell Technology:Basic and Applied Aspects vol.10, p.69-73(K itagawa, Y., Matsuda, T. and Iijima, S. eds.), Kluwer Academic Publishers, 1999; See Tomizuka, K. et. al., Proc. Natl. Acad. Sci. USA (2000) 97, p.722-727. Alternatively, a phage display selected from a human antibody library can be used. The antibody obtained by the method described in Wormstone, IM et al., Investigative Ophthalmology & Vis (2002)43 (7), p.2301-2308;Carmen, S. et. al., Briefings in Function al Genomics and Proteomics(2002), 1(2), p.189-203;Siriwardena, D. et. al., Ophth See, for example, Almology (2002) 109(3), p.427-431.

[0067] The antibody in the antibody-drug conjugate used in the present invention also includes modified antibodies. The modified form is an antibody according to the present invention that has been chemically or biologically modified. Chemical modifications include the attachment of chemical moieties to the amino acid backbone, N- or O-linkages. - Chemical modifications including the attachment of chemical moieties to the bound carbohydrate chain. The protein may include post-translational modifications (e.g., addition of N-linked or O-linked glycosylation, N- or C-terminal Processing, deamidation, isomerization of aspartic acid, oxidation of methionine, etc. By expressing this in a prokaryotic host cell, a methionine residue was added to the N-terminus. Also, in order to enable detection or isolation of the antibody or antigen according to the present invention, Such modifications include those labeled with an enzyme label, a fluorescent label, and an affinity label. Such modified antibodies according to the present invention are included in the meaning of the term "antibody." These are useful for improving affinity, reducing antigenicity, detecting or isolating antibodies or antigens, and the like.

[0068] In addition, it is possible to regulate the sugar chain modification (glycosylation, deglycosylation, etc.) of the antibody of the present invention. It is possible to enhance antibody-dependent cellular cytotoxicity by glycosylation of antibodies. Techniques for adjusting chain modification include those described in International Publication No. 99 / 54342 and International Publication No. 00 / 6173. 9, International Publication No. 02 / 31140, etc. are known, but are not limited to these. The antibodies according to the present invention also include those in which the glycosylation has been regulated.

[0069] In addition, antibodies produced in cultured mammalian cells have a lysine residue at the carboxyl terminus of the heavy chain. It is known that the nucleotide group is deleted (Journal of Chromatography A, 705: 129-134 (1995)). In addition, two amino acid residues, glycine and lysine, were deleted from the carboxyl terminus of the heavy chain, resulting in a new It is known that proline residues located at the carboxyl terminus are amidated (Ana Lytical Biochemistry, 360: 75-83(2007)). However, deletions and modifications of these heavy chain sequences The antigen-binding ability and effector function of antibodies (complement activation, antibody-dependent cellular cytotoxicity, etc.) Therefore, the antibody of the present invention includes the modified antibody and and functional fragments of said antibodies, each of which has one or two amino acids at the carboxyl terminus of the heavy chain. The deleted fragments and the amidated fragments (e.g., the protease at the carboxyl terminal) However, the heavy chains may be modified so as to have the same antigen-binding ability and effector function as the heavy chains. The carboxyl-terminal deletions of the heavy chains of the antibodies of the present invention may be carried out in the above-mentioned species, so long as the function is maintained. The two heavy chains constituting the antibody of the present invention may be full-length or the above-mentioned deleted forms. The heavy chain may be any one of the heavy chains selected from the group consisting of The ratio of the amounts of each deletion may be determined by the mammalian cultured cell line producing the antibody of the present invention. Although the type of cell and culture conditions may affect the antibody of the present invention, the antibody preferably has two heavy chains. In both cases, one amino acid residue is deleted from the carboxyl terminus. Cut.

[0070] The isotype of the antibody according to the present invention may be, for example, IgG (IgG1, IgG2, Ig Examples of the antibody include IgG1 and IgG2. It is possible.

[0071] The antibody in the antibody-drug conjugate used in the present invention is not particularly limited. For example, For example, anti-HER2 antibody, anti-HER3 antibody, anti-TROP2 antibody, anti-B7-H3 antibody, anti-CD 3 antibodies, anti-CD30 antibody, anti-CD33 antibody, anti-CD37 antibody, anti-CD56 antibody, anti-CD9 8 antibody, anti-DR5 antibody, anti-EGFR antibody, anti-EPHA2 antibody, anti-FGFR2 antibody, anti-FG Examples of antibodies include FR4 antibodies, anti-FOLR1 antibodies, anti-VEGF antibodies, and anti-GPR20 antibodies. and preferably an anti-HER2 antibody, an anti-HER3 antibody, an anti-TROP2 antibody, an anti-B7-H3 antibody, and anti-GPR20 antibodies.

[0072] In the present invention, the term "anti-HER2 antibody" refers to an antibody that binds to HER2 (Human Epidermal Growth Factor (HER2) l Growth Factor Receptor Type 2; ErbB-2) Specifically binds to HER2, and preferably binds to HER2-expressing cells. The antibody has the activity of activating the IL-1 domain.

[0073] Examples of anti-HER2 antibodies include trastuzumab (US Patent No. 5821337) and Pertuzumab (International Publication No. No. 01 / 00245, and preferably trastuzumab. do.

[0074] In the present invention, "trastuzumab" refers to the amino acid sequence represented by amino acid number 1 in SEQ ID NO:1 (Figure 1). and a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO:2 (FIG. 2) A humanized anti-HER2 monoclonal antibody comprising a light chain consisting of an amino acid sequence set forth in Nos. 1 to 214. It is a clonal antibody.

[0075] In the present invention, the term "anti-HER3 antibody" refers to an antibody that binds to HER3 (Human Epidermal Growth Factor (HER3) l Growth Factor Receptor Type 3; ErbB-3) The antibody specifically binds to HER3 on the surface of a HER3-expressing cell, and preferably binds to the HER3 expression. The antibodies shown have the activity of being internalized into the present cells.

[0076] Examples of anti-HER3 antibodies include Patritumab (U3-1 287), U1-59 (International Publication No. 2007 / 077028), MM-121 (ser ibantumab), the anti-ERBB3 antibody described in WO 2008 / 100624, G-7116 (Lumretuzumab) and LJM-716 (Elgemtumab) b) can be mentioned, and preferably, can be mentioned patritumab and U1-59. do.

[0077] In the present invention, the term "anti-TROP2 antibody" refers to an antibody that specifically binds to TROP2 (TACSTD2: Tumor r-associated calcium signal transducer 2 ; EGP-1) and preferably binds to TROP2, thereby 1 shows antibodies having the activity of being internalized into ROP2-expressing cells.

[0078] As an anti-TROP2 antibody, for example, hTINA1-H1L1 (International Publication No. 2015 / 013366) No. 098099).

[0079] In the present invention, the term "anti-B7-H3 antibody" refers to a B7-H3 (B cell antibody PD-L3; CD276; and Preferably, the antibody has an activity of being internalized into B7-H3 expressing cells by binding to B7-H3. The antibodies shown are:

[0080] As an anti-B7-H3 antibody, for example, M30-H1-L4 (International Publication No. 2014 / 05 No. 7687).

[0081] In the present invention, the term "anti-GPR20 antibody" refers to an antibody that binds to GPR20 (G Protein-CoA). Specifically binds to GPR20, preferably GPR20. The figures show antibodies that have the activity of being internalized into GPR20-expressing cells when combined with GPR20. As an anti-GPR20 antibody, for example, h046-H4e / L7 (International Publication No. 2018 / 1 No. 35501).

[0082] [Drug-Linker Intermediates Used in the Preparation of Antibody-Drug Conjugates] The drug linker intermediate used in the production of the antibody-drug conjugate of the present invention has the following formula: As shown.

[0083] [ka]

[0084] The above drug linker intermediates can be prepared according to the methods described in WO 2014 / 057687 and WO 2014 / 057687. 15 / 098099, WO 2015 / 115091, and WO 2015 The present invention can be manufactured by referring to the description in US Pat. No. 5,998,998, etc.

[0085] [Conjugation of antibodies and drug-linker intermediates] The antibody-drug conjugate used in the present invention comprises the above-mentioned drug linker intermediate and a thiol linker intermediate. It is produced by reacting an antibody having a carboxyl group (also called a sulfhydryl group) with the It is possible.

[0086] Antibodies bearing sulfhydryl groups can be obtained by methods well known in the art (Hermanson, G. T, Bioconjugate Techniques, pp.56-136, pp.456-493, Academic Press (1996)). example For example, a reducing agent such as tris(2-carboxyethyl)phosphine hydrochloride (TCEP) can be used as an anti- 0.3 to 3 molar equivalents per intracellular interchain disulfide are used, and ethylenediamine By reacting with the antibody in a buffer containing a chelating agent such as EDTA, To obtain an antibody having a sulfhydryl group in which the interchain disulfide has been partially or completely reduced It is possible.

[0087] In addition, 2 to 20 molar equivalents of a drug linker per antibody having a sulfhydryl group are - Using the intermediate, antibody-drug conjugates with 2 to 8 drugs bound to each antibody are prepared. Gates can be manufactured.

[0088] The average number of drugs bound per antibody molecule of the produced antibody-drug conjugate can be calculated using, for example, For example, the antibody-drug conjugate and its conjugate at two wavelengths of 280 nm and 370 nm The UV method is a method of calculating the UV absorbance of the jugation precursor (UV method), and the The drug-substance conjugate was treated with a reducing agent, and each of the resulting fragments was analyzed by HPLC. This can be done by a quantitative determination and calculation method (HPLC method).

[0089] Conjugation of the antibody to the drug linker intermediate and the antibody-drug conjugate The average number of drugs bound per molecule was calculated according to the method described in WO 2014 / 057687 and WO 2014 / 057687. Publication No. 2015 / 098099, International Publication No. 2015 / 115091, International Publication No. The present invention was implemented with reference to the descriptions in International Publication No. 2018 / 135501 and International Publication No. 015 / 155998. This can be done.

[0090] In the present invention, the term "anti-HER2 antibody-drug conjugate" refers to an antibody-drug conjugate. Antibody-drug conjugates in which the antibody in the gate is an anti-HER2 antibody are shown.

[0091] The anti-HER2 antibody is preferably a polypeptide comprising the amino acid sequence set forth in SEQ ID NO:1, the amino acid sequence set forth in SEQ ID NO:1 to 449. A heavy chain consisting of the amino acid sequence of SEQ ID NO:2 and an amino acid sequence as set forth in SEQ ID NO:2, or an antibody comprising a light chain consisting of the amino acid sequence set forth in SEQ ID NO:1. and a light chain consisting of the amino acid sequence set forth in SEQ ID NO:2.

[0092] Drug per antibody of the anti-HER2 antibody-drug conjugate used in the present invention The average number of bonds in the linker is preferably 2 to 8, more preferably 3 to 8, and More preferably, it is 7 to 8, even more preferably, it is 7.5 to 8, and even more preferably, it is It is about 8.

[0093] The anti-HER2 antibody-drug conjugate used in the present invention is The present invention can be manufactured by referring to the description in US Pat. No. 5 / 115091.

[0094] In the present invention, the term "anti-HER3 antibody-drug conjugate" refers to an antibody-drug conjugate. Antibody-drug conjugates in which the antibody in the gate is an anti-HER3 antibody are shown.

[0095] The anti-HER3 antibody preferably has a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO: 3 and a An antibody comprising a light chain consisting of the amino acid sequence set forth in No. 4, or a heavy chain carbohydrate of said antibody. This is an antibody that lacks the xyl-terminal lysine residue.

[0096] Drug per antibody of the anti-HER3 antibody-drug conjugate used in the present invention The average number of bonds in the linker is preferably 2 to 8, more preferably 3 to 8, and More preferably, it is 7 to 8, even more preferably, it is 7.5 to 8, and even more preferably, it is It is about 8.

[0097] The anti-HER3 antibody-drug conjugate used in the present invention is The present invention can be produced by referring to the description in US Pat. No. 5 / 155998.

[0098] In the present invention, the term "anti-TROP2 antibody-drug conjugate" refers to an antibody-drug conjugate. 1 shows an antibody-drug conjugate in which the antibody in the conjugate is an anti-TROP2 antibody.

[0099] The anti-TROP2 antibody preferably has a sequence similar to that shown in SEQ ID NO:5 at amino acid numbers 20 to 470. and a heavy chain having the amino acid sequence set forth in SEQ ID NO:6, which is represented by amino acid numbers 21 to 234. or the carboxyl terminus of the heavy chain of said antibody, It is an antibody in which the lysine residues are deleted.

[0100] Drug per antibody of the anti-TROP2 antibody-drug conjugate used in the present invention The average number of bonds in the polymer linker is preferably 2 to 8, more preferably 3 to 5, and More preferably, it is from 3.5 to 4.5, and even more preferably, it is about 4.

[0101] The anti-TROP2 antibody-drug conjugate used in the present invention is The present invention can be produced by referring to the description in US Pat. No. 5,999,158.

[0102] In the present invention, the term "anti-B7-H3 antibody-drug conjugate" refers to an antibody-drug conjugate. 1 shows an antibody-drug conjugate in which the antibody in the conjugate is an anti-B7-H3 antibody.

[0103] The anti-B7-H3 antibody preferably comprises the sequence represented by amino acid numbers 20 to 471 in SEQ ID NO:7. and a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO:8, which is represented by amino acid numbers 21 to 233. or the carboxyl terminus of the heavy chain of said antibody, It is an antibody in which the lysine residues are deleted.

[0104] Drug per antibody of the anti-B7-H3 antibody-drug conjugate used in the present invention The average number of bonds in the polymer linker is preferably 2 to 8, more preferably 3 to 5, and More preferably, it is from 3.5 to 4.5, and even more preferably, it is about 4.

[0105] The anti-B7-H3 antibody-drug conjugate used in the present invention is The present invention can be produced by referring to the description in US Pat. No. 5,999,333, issued Dec. 14, 2004, and the like.

[0106] In the present invention, the term "anti-GPR20 antibody-drug conjugate" refers to an antibody-drug conjugate. 1 shows an antibody-drug conjugate in which the antibody in the conjugate is an anti-GPR20 antibody.

[0107] The anti-GPR20 antibody preferably has a sequence similar to that shown in SEQ ID NO:9 at amino acid numbers 20 to 472. and a heavy chain consisting of the amino acid sequence set forth above, and a heavy chain consisting of amino acid numbers 21 to 234 in SEQ ID NO:10. An antibody comprising a light chain having the amino acid sequence described above, or the heavy chain carboxyl terminus of said antibody. This is an antibody that has deleted lysine residues at its ends.

[0108] Drug per antibody of the anti-GPR20 antibody-drug conjugate used in the present invention The average number of bonds in the polymer linker is preferably 2 to 8, more preferably 3 to 8, and More preferably, it is 7 to 8, even more preferably, it is 7.5 to 8, and even more preferably, it is is about 8.

[0109] The anti-GPR20 antibody-drug conjugate used in the present invention is The present invention can be produced by referring to the description in US Pat. No. 5,999,333, issued Dec. 18, 2001, and the like.

[0110] [Pharmaceutical composition] The pharmaceutical composition of the present invention will now be described. The pharmaceutical composition of the present invention comprises: (i) antibody-drug conjugates, (ii) a histidine buffer; (iii) sucrose or trehalose, and (iv) surfactants, A pharmaceutical composition comprising: (wherein the antibody-drug conjugate has the following formula:

[0111] [ka]

[0112] (wherein A represents the binding site to the antibody) The antibody-drug conjugate is formed by binding the drug linker represented by the formula: It is a Jugate.) In the present invention, the term "histidine buffer" refers to histidine and its salt (acid adduct). and preferably a mixture of L-histidine and L-histidine hydrochloride (and / or The water in which the histidine buffer of the present invention is dissolved is a mixture of histidine and hydrate. It is synonymous with thymidine buffer.

[0113] The amount of histidine buffer is preferably 3 to 20 mg per 20 mg of antibody-drug conjugate. 80 mmol, and more preferably 10 ~80 mmol, and even more preferably, per 20 mg of antibody-drug conjugate 10 to 40 mmol, and even more preferably, More preferably, the amount is 10 to 25 mmol per unit, and even more preferably, 20 mmol per unit, It is 10 or 25 mmol per gram.

[0114] The pH of the pharmaceutical composition of the present invention when dissolved in water is 100% in a histidine buffer. In other words, the histidine and histidine salt content depends on the histidine and histidine salt content. By adjusting the content of thiocyanate salt, a pharmaceutical agent that exhibits the desired pH when dissolved in water can be obtained. A pharmaceutical composition can be provided.

[0115] The pharmaceutical composition of the present invention is a composition in which the antibody-drug conjugate is dissolved in water at a concentration of 20 mg / mL. The pH in this state is preferably 4.0 to 7.0, and more preferably 5.0 to 6. 0. In addition, depending on the type of antibody in the antibody-drug conjugate, a more suitable p H can be selected. The amount of sucrose or trehalose is preferably about 100 mg per 20 mg of antibody-drug conjugate. In the case of sucrose, the amount is preferably 90 mg. In the case of trehalose, the amount is more preferably calculated as trehalose hydrate. Preferably, the amount is 100 mg. The pharmaceutical composition of the present invention also contains sucrose and trehalose. Of these, sucrose is more preferably used. In the present invention, a "surfactant" refers to a component of a pharmaceutical preparation that has a hydrophilic group and a hydrophobic group. The surfactant in the present invention is preferably a polysol. Polysorbate 80 (Tween 80), Polysorbate 20 (Tween 20 ), and polysorbate 60 (Tween 60), polyoxyethylene (160 ) Polyoxypropylene (30) glycol, polyoxyethylene hydrogenated castor oil 60, poly Polyoxyethylene castor oil, or sodium lauryl sulfate, and more preferably polyiso Polysorbate 80, or Polysorbate 20.

[0116] The amount of polysorbate 80 or polysorbate 20 is preferably adjusted to The amount of the glycerin is 0.05 to 1.6 mg per 20 mg of glycerin, and more preferably 0.1 to 1.6 mg. More preferably, the amount is 0.2 to 0.4 mg, and even more preferably, the amount is 0.2 to 0. .3 mg, and even more preferably 0.2 or 0.3 mg. Considering the above comprehensively, the pharmaceutical composition of the present invention is preferably (i) per 20 mg of antibody-drug conjugate, (ii) 3 to 80 mmol of a histidine buffer; (iii) 24 to 320 mg of sucrose or trehalose, and (iv) 0.05 to 1.6 mg of polysorbate 80 or polysorbate 20, A pharmaceutical composition comprising: More preferably, (i) per 20 mg of antibody-drug conjugate, (ii) 10 to 40 mmol of histidine buffer; (iii) 90 mg of sucrose or 100 mg of trehalose hydrate, and (iv) 0.2 to 0.4 mg of polysorbate 80 or polysorbate 20, A pharmaceutical composition comprising: Even more preferably, (i) per 20 mg of antibody-drug conjugate, (ii) 10 or 25 mmol of histidine buffer; (iii) 90 mg of sucrose, and (iv) 0.2 or 0.3 mg of polysorbate 80 or polysorbate 20, A pharmaceutical composition comprising: The pharmaceutical composition is an aqueous solution of the antibody-drug conjugate at a concentration of 20 mg / mL. can also be expressed as follows:

[0117] That is, the pharmaceutical composition of the present invention is preferably (i) 20 mg / mL of antibody-drug conjugate; (ii) a histidine buffer; (iii) sucrose or trehalose, (iv) a surfactant, and (v) water A pharmaceutical composition comprising: More preferably, (i) 20 mg / mL of antibody-drug conjugate; (ii) 3 to 80 mM histidine buffer; (iii) 2.4 to 32% sucrose or trehalose; (iv) 0.005 to 0.16% polysorbate 80 or polysorbate 20, and (v) water A pharmaceutical composition comprising: Even more preferably, (i) 20 mg / mL of antibody-drug conjugate; (ii) 10 to 40 mM histidine buffer; (iii) 9% sucrose or 10% trehalose hydrate; (iv) 0.02 to 0.04% polysorbate 80 or polysorbate 20, and (v) water A pharmaceutical composition comprising: Even more preferably, (i) 20 mg / mL of antibody-drug conjugate; (ii) 10 or 25 mM histidine buffer; (iii) 9% sucrose, and (iv) 0.02 or 0.03% polysorbate 80 or polysorbate 20, and (v) water A pharmaceutical composition comprising:

[0118] In the present invention, the percentages of sucrose and trehalose are based on the content per 1 ml of water. The percentage by weight is expressed in terms of the amount of sucrose per mL of water. For example, "9% sucrose" is "10% trehalose hydrate" means that it contains 90 mg of sucrose. This means that 100 mg of trehalose hydrate is contained per mL of water.

[0119] Similarly, in the present invention, the content of polysorbate 80 and polysorbate 20 "%" also indicates the weight percent relative to 1 mL of water. For example, "0.03% polysorbate "Polysorbate 80" contains 0.3 mg of polysorbate 80 per 1 mL of water. "0.03% polysorbate 20" means 0.3 mg of polysorbate 20 per mL of water. This means that Rubet 20 is included.

[0120] The pharmaceutical composition of the present invention may be more effectively used depending on the type of antibody in the antibody-drug conjugate. A suitable formulation can be selected.

[0121] For example, the antibody in the antibody-drug conjugate is an anti-HER2 antibody (preferably having the sequence A heavy chain having the amino acid sequence set forth in SEQ ID NO:1 to SEQ ID NO:449 and 2. An antibody, or a heavy chain having the amino acid sequence set forth in SEQ ID NO: 1 and an amino acid sequence set forth in SEQ ID NO: 2 In the case of an antibody having a light chain consisting of the amino acid sequence of teeth, (i) per 20 mg of antibody-drug conjugate, (ii) 25 mmol of histidine buffer; (iii) 90 mg of sucrose, and (iv) 0.3 mg of polysorbate 80, A pharmaceutical composition comprising: Furthermore, the antibody-drug conjugate is dissolved in water at a concentration of 20 mg / mL. The pH of the pharmaceutical composition is preferably 5.3 to 5.7, and more preferably 5.4 to 5. 6, and even more preferably 5.5.

[0122] The histidine buffer is expressed as the content of L-histidine and L-histidine hydrochloride. Therefore, the above pharmaceutical composition at pH 5.5 has the following properties: (i) per 20 mg of antibody-drug conjugate, (ii) 0.89 mg L-histidine, and 4.04 mg L-histidine hydrochloride hydrate thing, (iii) 90 mg of sucrose; and (iv) It can be expressed as a pharmaceutical composition comprising 0.3 mg of polysorbate 80.

[0123] Furthermore, when expressed as a unit formulation containing 100 mg of the antibody-drug conjugate, pH is 5.5, (i) 100 mg of the antibody-drug conjugate; (ii) 4.45 mg L-histidine, and 20.2 mg L-histidine hydrochloride hydrate thing, (iii) 450 mg of sucrose; and (iv) It can be expressed as a pharmaceutical composition comprising 1.5 mg of polysorbate 80.

[0124] Alternatively, the antibody-drug conjugate is expressed as an aqueous solution having a concentration of 20 mg / mL. and the pharmaceutical composition comprises: (i) 20 mg / mL of antibody-drug conjugate; (ii) 25 mM histidine buffer; (iii) 9% sucrose, and (iv) 0.03% polysorbate 80, and (v) water and having a pH of 5.5.

[0125] The antibody in the antibody-drug conjugate is an anti-HER3 antibody (preferably, and a light chain consisting of the amino acid sequence set forth in SEQ ID NO:4. or an antibody having a deletion of a lysine residue at the carboxyl terminus of the heavy chain of said antibody) In the latter case, the pharmaceutical composition of the present invention preferably comprises (i) per 20 mg of antibody-drug conjugate, (ii) 25 mmol of histidine buffer; (iii) 90 mg of sucrose, and (iv) 0.3 mg of polysorbate 20, A pharmaceutical composition comprising: Furthermore, the antibody-drug conjugate is dissolved in water at a concentration of 20 mg / mL. The pH of the pharmaceutical composition is preferably 5.2 to 5.6, and more preferably 5.3 to 5. 5, and even more preferably 5.4.

[0126] The histidine buffer is expressed as the content of L-histidine and L-histidine hydrochloride. In other words, the pharmaceutical composition at a pH of 5.4 is preferably (i) per 20 mg of antibody-drug conjugate, (ii) 0.81 mg L-histidine, and 4.14 mg L-histidine hydrochloride hydrate thing, (iii) 90 mg of sucrose; and (iv) It can be expressed as a pharmaceutical composition comprising 0.3 mg of polysorbate 20.

[0127] Furthermore, when expressed as a unit formulation containing 100 mg of the antibody-drug conjugate, pH is 5.4, (i) 100 mg of the antibody-drug conjugate; (ii) 4.06 mg L-histidine, and 20.7 mg L-histidine hydrochloride hydrate thing, (iii) 450 mg of sucrose; and (iv) It can be expressed as a pharmaceutical composition comprising 1.5 mg of polysorbate 20.

[0128] Alternatively, the antibody-drug conjugate is expressed as an aqueous solution having a concentration of 20 mg / mL. and the pharmaceutical composition comprises: (i) 20 mg / mL of antibody-drug conjugate; (ii) 25 mM histidine buffer; (iii) 9% sucrose, and (iv) 0.03% polysorbate 20, and (v) water and having a pH of 5.4.

[0129] The antibody in the antibody-drug conjugate is an anti-TROP2 antibody (preferably, SEQ ID NO:5 and a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO: 20 to 470 of SEQ ID NO: 6. an antibody comprising a light chain having an amino acid sequence as set forth in amino acid numbers 21 to 234 of or an antibody lacking a lysine residue at the carboxyl terminus of the heavy chain of the antibody) The pharmaceutical composition of the present invention preferably comprises: (i) per 20 mg of antibody-drug conjugate, (ii) 10 mmol of histidine buffer; (iii) 90 mg of sucrose, and (iv) 0.2 or 0.3 mg of polysorbate 80, A pharmaceutical composition comprising: Furthermore, the antibody-drug conjugate is dissolved in water at a concentration of 20 mg / mL. The pH of the pharmaceutical composition is preferably 5.8 to 6.2, and more preferably 5.9 to 6. 1, and even more preferably 6.0.

[0130] The histidine buffer is expressed as the content of L-histidine and L-histidine hydrochloride. In other words, the pharmaceutical composition at a pH of 6.0 is preferably (i) per 20 mg of antibody-drug conjugate, (ii) 0.78 mg L-histidine, and 1.05 mg L-histidine hydrochloride hydrate thing, (iii) 90 mg of sucrose; and (iv) Expressed as a pharmaceutical composition containing 0.2 or 0.3 mg of polysorbate 80. can be done.

[0131] When expressed as a unit formulation containing 100 mg of antibody-drug conjugate, the pH is 6.0 The pharmaceutical composition comprises (i) 100 mg of the antibody-drug conjugate; (ii) 3.88 mg L-histidine, and 5.26 mg L-histidine hydrochloride hydrate thing, (iii) 450 mg of sucrose; and (iv) Expressed as a pharmaceutical composition containing 1.0 or 1.5 mg of polysorbate 80. can be done.

[0132] Alternatively, the antibody-drug conjugate is expressed as an aqueous solution having a concentration of 20 mg / mL. and the pharmaceutical composition comprises: (i) 20 mg / mL of antibody-drug conjugate; (ii) 10 mM histidine buffer; (iii) 9% sucrose, and (iv) 0.02 or 0.03% polysorbate 80, and (v) water and having a pH of 6.0.

[0133] The antibody in the antibody-drug conjugate is an anti-B7-H3 antibody (preferably, SEQ ID NO: 7 and a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO: 20 to 471 in an antibody having a light chain consisting of the amino acid sequence set forth in amino acid numbers 21 to 233 in or an antibody lacking a lysine residue at the carboxyl terminus of the heavy chain of the antibody) The pharmaceutical composition of the present invention preferably comprises: (i) per 20 mg of antibody-drug conjugate, (ii) 10 mmol of histidine buffer; (iii) 90 mg of sucrose, and (iv) 0.2 or 0.3 mg of polysorbate 20, A pharmaceutical composition comprising: Furthermore, the antibody-drug conjugate is dissolved in water at a concentration of 20 mg / mL. The pH of the pharmaceutical composition is preferably 5.7 to 6.1, and more preferably 5.8 to 6. 0, and even more preferably 5.9.

[0134] The histidine buffer is expressed as the content of L-histidine and L-histidine hydrochloride. In this case, the pharmaceutical composition at a pH of 5.9 is preferably (i) per 20 mg of antibody-drug conjugate, (ii) 0.65 mg L-histidine, and 1.22 mg L-histidine hydrochloride hydrate thing, (iii) 90 mg of sucrose; and (iv) Expressed as a pharmaceutical composition containing 0.2 or 0.3 mg of polysorbate 20. can be done.

[0135] Furthermore, when expressed as a unit formulation containing 100 mg of the antibody-drug conjugate, pH is 5.9, (i) 100 mg of the antibody-drug conjugate; (ii) 3.23 mg L-histidine, and 6.12 mg L-histidine hydrochloride hydrate thing, (iii) 450 mg of sucrose; and (iv) Expressed as a pharmaceutical composition containing 1.0 or 1.5 mg of polysorbate 20. can be done.

[0136] Alternatively, the antibody-drug conjugate is expressed as an aqueous solution having a concentration of 20 mg / mL. and the pharmaceutical composition comprises: (i) 20 mg / mL of antibody-drug conjugate; (ii) 10 mM histidine buffer; (iii) 9% sucrose, and (iv) 0.02 or 0.03% polysorbate 20, and (v) water and having a pH of 5.9.

[0137] The antibody in the antibody-drug conjugate is an anti-GPR20 antibody (preferably, SEQ ID NO: 9 and a heavy chain consisting of the amino acid sequence set forth in amino acid numbers 20 to 472 of SEQ ID NO: 1. 0, comprising a light chain consisting of the amino acid sequence set forth in amino acid numbers 21 to 234. antibody, or an antibody lacking a lysine residue at the carboxyl terminus of the heavy chain of the antibody) In this case, the pharmaceutical composition of the present invention is preferably (i) per 20 mg of antibody-drug conjugate, (ii) 10 mmol of histidine buffer; (iii) 90 mg of sucrose, and (iv) 0.3 mg of polysorbate 80, A pharmaceutical composition comprising: Furthermore, the antibody-drug conjugate is dissolved in water at a concentration of 20 mg / mL. The pH of the pharmaceutical composition is preferably 5.2 to 5.6, and more preferably 5.3 to 5. 5, and even more preferably 5.4.

[0138] The histidine buffer is expressed as the content of L-histidine and L-histidine hydrochloride. In other words, the pharmaceutical composition at a pH of 5.4 is preferably (i) per 20 mg of antibody-drug conjugate, (ii) 0.32 mg L-histidine, and 1.66 mg L-histidine hydrochloride hydrate thing, (iii) 90 mg of sucrose; and (iv) It can be expressed as a pharmaceutical composition comprising 0.3 mg of polysorbate 80.

[0139] Furthermore, when expressed as a unit formulation containing 100 mg of the antibody-drug conjugate, pH is 5.4, (i) 100 mg of the antibody-drug conjugate; (ii) 1.62 mg L-histidine, and 8.29 mg L-histidine hydrochloride hydrate thing, (iii) 450 mg of sucrose; and (iv) It can be expressed as a pharmaceutical composition comprising 1.5 mg of polysorbate 80.

[0140] Alternatively, the antibody-drug conjugate is expressed as an aqueous solution having a concentration of 20 mg / mL. and the pharmaceutical composition comprises: (i) 20 mg / mL of antibody-drug conjugate; (ii) 10 mM histidine buffer; (iii) 9% sucrose, and (iv) 0.03% polysorbate 80, and (v) water and having a pH of 5.4.

[0141] The pharmaceutical composition of the present invention is preferably an injection, more preferably an aqueous injection or a frozen It is preferably a lyophilized injection, and even more preferably a lyophilized injection.

[0142] In the present invention, an "injection" refers to a drug that is administered by injection into living tissue (e.g., intravenously, intradermally, The present invention also relates to a pharmaceutical composition for administration via the intravenous route (subcutaneous, intramuscular, intraperitoneal, etc.). This includes not only liquid injections, but also solid injections that are dissolved and turned into a liquid when used. In the case of a liquid injection, the solvent may be water or a solvent other than water. A mixed solvent of water and a solvent other than water may also be used.

[0143] In the present invention, the term "aqueous injection" refers to a liquid injection, the solvent of which is water (preferably an injection This refers to a pharmaceutical composition that is In the present invention, a "lyophilized injection" is a drug that is dissolved in water (preferably water for injection) when used. The solid injection can be prepared by freeze-drying an aqueous solution containing a predetermined amount of a medicinal ingredient. The present invention refers to a pharmaceutical composition obtained by When the pharmaceutical composition of the present invention is an aqueous injection, the concentration of the antibody-drug conjugate is preferably Preferably, the concentration is 5 to 60 mg / mL, more preferably, 15 to 40 mg / mL, and further preferably, More preferably, it is 20 mg / mL. When the pharmaceutical composition of the present invention is an aqueous injection, the pharmaceutical composition of the present invention is preferably frozen. The data can be stored in a compressed state.

[0144] When the pharmaceutical composition of the present invention is a lyophilized injection, the pharmaceutical composition of the present invention is preferably It can be stored in an amber vial.

[0145] When the pharmaceutical composition of the present invention is a freeze-dried injection, the pharmaceutical composition of the present invention and water for injection are mixed together. A kit containing the water for injection can be preferably used. The information can be stored in a file.

[0146] Lyophilization of the antibody-drug conjugate used in the present invention with a content of 20 mg The injection can be suitably used by reconstituting it with 1 mL of water for injection. In addition, when the content of the antibody-drug conjugate used in the present invention is 100 mg, The lyophilized injection preparation should be reconstituted with 5 mL of water for injection before use. can be done.

[0147] When the pharmaceutical composition of the present invention is a freeze-dried injection, the method for producing the pharmaceutical composition of the present invention is , preferably (1) A specified amount of (i) antibody-drug conjugates, (ii) a histidine buffer; (iii) sucrose or trehalose, and (iv) surfactants, preparing an aqueous solution comprising (2) if necessary, adjusting the pH of the aqueous solution to a predetermined value, and then (3) freeze-drying the aqueous solution; The manufacturing method includes: The step of freeze-drying the aqueous solution is preferably followed by annealing at a shelf temperature near the eutectic point of the aqueous solution. Here, "near the eutectic point" refers to a process in which the temperature is 1.5°C lower than the eutectic point and the temperature is increased to the eutectic point. This indicates a range of temperatures 1.5°C higher than the point. In the present invention, "annealing" refers to freezing at a product temperature above the freezing glass transition point (Tg'). This refers to the process of growing the ice crystals in the solid. As the ice crystals grow larger, they sublimate during drying. This reduces the resistance to sublimation and increases the size of the freeze-dried cake. It is expected that the shape of the product will improve and the drying time will be shortened. In the present invention, the term "shelf temperature" refers to the temperature inside the apparatus (system) in which freeze-drying is performed. In the present invention, the term "product temperature" refers to the temperature of the object (product) to be freeze-dried. In the present invention, the term "freeze-dried cake" refers to a freeze-dried cake obtained by a series of freeze-drying processes. It means a sintered, dried body (a solid with a porous structure). As mentioned above, annealing is performed at a temperature 1.5°C lower than the eutectic point of the aqueous solution, and then at a temperature 1.5°C higher than the eutectic point. The reaction can be preferably carried out at a temperature in the range of 1.5° C. higher than the eutectic point of the aqueous solution, and more preferably at a temperature in the range of 1.5° C. higher than the eutectic point of the aqueous solution. The heating can be performed at the same shelf temperature. These manufacturing methods preferably include annealing at a shelf temperature 5° C. below the eutectic point. In comparison with the above, the time required for the primary drying process is shortened. The shrinkage was smaller than that of the case where the shelf temperature was 5°C lower than the eutectic point. The freeze-dried cake obtained is characterized by a low

[0148] In the case of the pharmaceutical composition of the present invention, the eutectic point is in the range of about -3°C to about -1°C. The drying can be preferably carried out at a shelf temperature of -4.5°C to 0.5°C, more preferably at - It can be carried out at a shelf temperature of 4 to -1°C, and even more preferably at a shelf temperature of -4 to -2°C. More preferably, the shelf temperature is between -3 and -2°C. can be performed at a shelf temperature of -2.5°C.

[0149] After annealing, the pharmaceutical composition of the present invention is subjected to a primary drying process and a secondary drying process. Through this process, it is manufactured into a freeze-dried injection.

[0150] In the present invention, the "primary drying process" refers to one of the steps in the production of a freeze-dried injection, It refers to a process for sublimating free water in a frozen body. In the present invention, the "secondary drying process" is one of the processes for producing freeze-dried injections, The term "bound water" refers to a process for sublimating water bound to a substance (bound water).

[0151] The primary drying process is preferably carried out at a shelf temperature of -5 to 5°C and under a vacuum of 5 to 15 Pa. More preferably, the reaction can be carried out at a shelf temperature of -4 to 4°C and under a vacuum of 7 to 13 Pa. and more preferably, the reaction can be carried out at a shelf temperature of -3 to 3°C and under a vacuum of 8 to 12 Pa. and more preferably, the reaction can be carried out at a shelf temperature of -2 to 2°C and under a vacuum of 9 to 11 Pa. and more preferably, the reaction can be carried out at a shelf temperature of -1 to 1°C and under a vacuum of 9 to 11 Pa. and even more preferably at a shelf temperature of 0° C. and under a vacuum of about 10 Pa.

[0152] The secondary drying process is preferably carried out at a shelf temperature of 40 to 50° C. and under a vacuum of 5 to 15 Pa or It can be carried out without vacuum, more preferably at a shelf temperature of 42 to 48°C and a true pressure of 7 to 13 Pa. The heating can be carried out under atmospheric conditions, and more preferably at a shelf temperature of 44 to 46° C. and a true pressure of 9 to 11 Pa. The reaction can be carried out under vacuum, and even more preferably under a shelf temperature of 45° C. and a vacuum of 10 Pa. It is possible.

[0153] The quality of the pharmaceutical composition of the present invention can be evaluated, for example, at 50° C., 40° C. / 75% RH, 25° C. / 6 Under 0% RH or 5℃ conditions, for 1 month, 3 months, 6 months, 12 months, Storage stability for months, 18 months, 24 months, or 36 months was measured using protein concentration, Moisture, insoluble particulates, DAR, monomer / aggregate / fragment ratio, NPI, IoP, charge difference The degree of adhesion can be confirmed by evaluating the antibody-drug complex using the properties and pH as indicators. Depending on the type of antibody in the conjugate, binding activity against the antigen is Quality evaluation can also be performed using indicators such as ty and potency (bioassay). In addition, quality evaluation based on drug distribution and rCE-SDS, Quality evaluation can also be performed by observing the properties (appearance). In the case of aqueous injections, the osmotic pressure ratio is In the case of freeze-dried injections, the reconstitution time can be used as an index for quality evaluation. These quality evaluations can be used to evaluate the superiority of the compound over existing pharmaceutical compositions. You can also check the position. The freeze-drying method of the present invention also includes the antibody-drug conjugate used in the present invention. General application to pharmaceutical compositions as well as aqueous solutions containing sucrose or trehalose is expected. Therefore, annealing at a shelf temperature near the eutectic point of an aqueous solution containing sucrose or trehalose is performed. A method for producing a freeze-dried injection preparation, comprising a process of suspending the freeze-dried injection solution in a eutectic solution. The temperature range is from 1.5°C lower than the eutectic point to 1.5°C higher than the eutectic point. falls within the scope of the invention. The annealing is preferably carried out in the presence of an aqueous solution containing sucrose or trehalose. This can be done at the same shelf temperature as the melting point. These manufacturing methods preferably include annealing at a shelf temperature 5° C. below the eutectic point. This method is characterized by the fact that the time required for the primary drying process is shortened compared to the conventional method.

[0154] In addition, these manufacturing methods preferably include annealing at a shelf temperature 5° C. lower than the eutectic point. The freeze-dried cake has less shrinkage than that obtained by the conventional freeze-dried method. do.

[0155] [Use of pharmaceutical composition] The pharmaceutical composition of the present invention can be applied to patients as a systemic therapy, and also can be applied locally to cancer tissues. It can be expected to have a therapeutic effect when applied to the following:

[0156] The pharmaceutical composition of the present invention can be preferably used for mammals, and more preferably can be used in humans.

[0157] Routes of introduction that can be used to administer the pharmaceutical composition of the present invention include, for example, intravenous The routes include intradermal, subcutaneous, intramuscular, and intraperitoneal, but preferably intravenous. be.

[0158] When the pharmaceutical composition of the present invention is an aqueous injection, it is preferably diluted with a suitable diluent and then: It can be administered intravenously by drip infusion. The diluent may be, for example, a glucose solution (preferably Examples of suitable solutions include 5% glucose solution and saline solution.

[0159] When the pharmaceutical composition of the present invention is a freeze-dried injection, it is dissolved in water (preferably water for injection). After dissolving, the required amount can be diluted with an appropriate diluent and then administered intravenously. Examples of the dilution solution include a glucose solution (preferably a 5% glucose solution) and physiological saline. It can be done.

[0160] The pharmaceutical composition of the present invention is characterized by the affinity of the antibody-drug conjugate of the present invention to an antigen, In other words, in terms of the dissociation constant (Kd value) for the antigen, the affinity is high (the Kd value is low). Therefore, the pharmaceutical effect can be obtained even at a small dose. The dosage of the composition can also be determined based on the affinity with the antigen. When the pharmaceutical composition is administered to humans, for example, the antibody-drug conjugate is administered in an amount of about 0.001 to 100 mg / kg (here, "mg / kg" means per kg of human body weight) (meaning the dose of the antibody-drug conjugate) once or once every 1 to 180 days For example, the antibody-drug conjugate used in the present invention may be administered multiple times at intervals. In the case of an anti-HER2 antibody-drug conjugate, the concentration is preferably 0.8 mg / kg to 8 mg A more preferred method is to administer 0.8 mg / kg once every three weeks. , 1.6mg / kg, 3.2mg / kg, 5.4mg / kg, 6.4mg / kg, 7.4 and a method of administering 10 mg / kg or 8 mg / kg once every 3 weeks. Preferably, 5.4 mg / kg, 6.4 mg / kg, 7.4 mg / kg, or 8 mg / kg. More preferably, the dose is 5.4 mg / kg, or Alternatively, 6.4 mg / kg may be administered once every three weeks.

[0161] The pharmaceutical composition of the present invention can be used for the treatment of cancer, preferably breast cancer, Stomach cancer (sometimes called gastric adenocarcinoma), colon cancer (sometimes called colorectal cancer, colon cancer, colon cancer, and rectal cancer), lung cancer (including small cell lung cancer and non-small cell lung cancer), esophageal Salivary gland cancer, gastroesophageal junction adenocarcinoma, bile duct cancer, Paget's disease, pancreatic cancer, ovarian cancer , uterine carcinosarcoma, urothelial carcinoma, prostate cancer, bladder cancer, gastrointestinal stromal tumor, gastrointestinal stromal tumor , cervical cancer, squamous cell carcinoma, peritoneal cancer, liver cancer, hepatocellular carcinoma, colon cancer, rectal cancer, Endometrial cancer, uterine cancer, kidney cancer, vulvar cancer, thyroid cancer, penile cancer, leukemia, malignant Lymphoma, plasmacytoma, myeloma, neuroepithelial tumor, nerve sheath tumor, head and neck cancer, skin A small number of cancers selected from the group consisting of cancer, pharyngeal cancer, gallbladder cancer, bile duct cancer, mesothelioma, and sarcoma. For example, the present invention can be used to treat at least one cancer. When the antibody-drug conjugate is an anti-ER2 antibody-drug conjugate, more preferably are breast cancer, gastric cancer, colon cancer, non-small cell lung cancer, esophageal cancer, salivary gland cancer, and gastroesophageal junction cancer. Adenocarcinoma, bile duct carcinoma, Paget's disease, pancreatic cancer, ovarian cancer, and uterine carcinosarcoma It can be used for the treatment of at least one cancer selected from the following, more preferably , breast cancer, gastric cancer, colon cancer, non-small cell lung cancer, esophageal cancer, salivary gland cancer, gastroesophageal junction cancer At least one cancer selected from the group consisting of cancer, bile duct cancer, and Paget's disease It can be used for the treatment of breast cancer, stomach cancer, colon cancer, or can be used for the treatment of non-small cell lung cancer.

[0162] The pharmaceutical composition of the present invention is selected as a drug for drug therapy, which is a major method for treating cancer. The drug can be used selectively to slow the growth of cancer cells, inhibit their proliferation, and These effects can lead to the development of a new type of cancer treatment. It can relieve cancer symptoms, improve quality of life, and save the lives of cancer patients. Even if the destruction of cancer cells is not achieved, the inhibition of cancer cell proliferation and the control of cancer cells are achieved. Controls help cancer patients achieve longer survival with higher quality of life It can be done.

[0163] In addition to the use of the drug alone in such drug therapy, the pharmaceutical composition of the present invention can also be used as an adjuvant. It can also be used in combination with other therapies in a multicenter trial, such as surgery or radiation therapy. It can be combined with chemotherapy, hormonal therapy, etc. Furthermore, it can be used in neoadjuvant therapy. It can also be used as a drug therapy.

[0164] In addition to the above-mentioned therapeutic uses, the pharmaceutical composition of the present invention can suppress the proliferation of micrometastatic cancer cells. For example, it is expected that the effect of preventing the spread of cancer may be prevented by preventing or even destroying the cancer. Its effect of inhibiting and destroying cancer cells in body fluids and of destroying minute cancer cells immediately after implantation in any tissue. It is expected to have the effect of suppressing and destroying cancer cells. It is expected that this will have the effect of suppressing and preventing cancer metastasis after removal.

[0165] The pharmaceutical composition of the present invention can also be administered in combination with other cancer therapeutic agents, thereby It can enhance the antitumor effect. , 5-fluorouracil (5-FU), pertuzumab, trachea Trastuzumab, Paclitaxel, Cal Carboplatin, Cisplatin, Gemcitabine Gemcitabine, Capecitabine, Ili Irinotecan (CPT-11), Docetaxel l), Pemetrexed, Sorafenib , Vinblastin, Vinorelbine , Everolimus, Tanespimycin ), Bevacizumab, Oxaliplatin in), Lapatinib, Trastuzumab emtansine uzumab emtansine) (T-DM1) or International Publication No. 2003 / 0380 Drugs listed in issue 43, as well as LH-RH analogs (Leuprorelin in), Goserelin, etc.), Estramustine phosphate ( Estramustine Phosphate, estrogen antagonist (tamoxifen) Tamoxifen, Raloxifene, etc.), aromatase α-amyloidase inhibitors (anastrozole, letrozole) zole, exemestane, etc. There are no limitations as long as the drug has antitumor activity. EXAMPLES

[0166] The present invention will be described in detail with reference to the following examples, but the present invention is not limited thereto. Furthermore, these are not to be construed as limiting in any way.

[0167] [Example 1: Production of antibody-drug conjugate (1)] A humanized anti-HER2 antibody was produced based on the manufacturing method described in International Publication WO 2015 / 115091. Using the compound (trastuzumab),

[0168] [ka]

[0169] (wherein A represents the binding site to the antibody) and an anti-HER2 antibody linked via a thioether bond. -drug conjugate (hereinafter referred to as "antibody-drug conjugate (1)") The DAR of the antibody-drug conjugate (1) was 7.8.

[0170] [Example 2: Preparation of formulation of antibody-drug conjugate (1)] Preparation of the formulation (1) Antibody-drug conjugate (1) (20 mg / mL), 10 mM histidine buffer, 1 Aqueous solution containing 1.1% trehalose hydrate and 0.02% polysorbate 20 (pH 5 .8) Dialysis cassette (dialysis membrane, Slide-A-lyzer, MWCO 20,00 0) and the amount of the sample injected into the dialysis cassette was 50 times or more that of the excipient of the intended formulation. The dialysis was performed twice at 5°C for more than 5 hours. The solution was concentrated or diluted appropriately so that the concentration was about 20 mg / mL. The solution was filtered using a 2 μm filter. 1 mL of the filtered solution was filled into glass vials. The rubber stoppers were partially plugged and freeze-dried under the conditions shown in Table 1. After freeze-drying, the bottles with the rubber stoppers plugged were The vial was then capped. Filtration and filling were carried out in a clean bench.

[0171] [Table 1]

[0172] Preparation of the formulation (2) Antibody-drug conjugate (1) (20 mg / mL), 10 mM histidine buffer, 1 Aqueous solution containing 1.1% trehalose hydrate and 0.02% polysorbate 20 (pH 5 8) was dispensed into a UF membrane kit (AMICON ULTRA-15, 30 kDa) and diluted for 30 minutes. The concentrate was concentrated approximately twice by centrifugation at 100 rpm and 5°C. Porphyrin 20 was removed by adsorption onto a hydrophobic adsorbent (Amberlite XAD7HP). The drug solution with Resorbate 20 removed was placed in a dialysis cassette (dialysis membrane, Slide-A-lyzer , MWCO 20,000) and the amount of sample injected into the dialysis cassette was 50 times or more. Dialysis was performed against a buffer solution containing the desired formulation of excipients. Dialysis was performed twice at 5°C for more than 5 hours. Concentrate or dilute appropriately so that the protein concentration after dialysis is approximately 20 mg / mL. After that, polysorbate 20 or polysorbate 80 was added and mixed. The solution was filtered using a 0.22 μm filter. 1 mL of the filtered solution was poured into a glass vial. The tubes were filled with water, partially plugged with rubber stoppers, and freeze-dried under the conditions shown in Table 2. After freeze-drying, the rubber stoppers were plugged. The vials were then capped. Filtration and filling were carried out in a clean bench.

[0173] [Table 2]

[0174] [Example 3: Stability test] The freeze-dried injection was stored at 40°C / 75% RH for 3 months. Protein concentration (UV method) Water content (measured by Karl Fischer method), insoluble particles (Micro Flo w Imaging), Drug Antibody Ratio (DAR) ( HPLC method), and the ratio of monomers, aggregates, and fragments (SE-HPLC (SEC) Measurement), Non Proteinous Impurity (NPI) (HPLC method) Impurity of Payload (IoP) (measured by HPLC method) ), charge isomers (measured by CZE), pH, and HER2 binding activity The stability was compared and evaluated using the ity (measured by ELISA) as an indicator.

[0175] Example 4: Excipient Screening The excipient of the lyophilized injection has a proven track record of use in biopharmaceuticals, and is a concentrated solution with an almost isotonic osmotic pressure. The concentrations of trehalose hydrate, 10%, sucrose, and 5% sorbitol were selected. The other components were the same: antibody-drug conjugate (1) (20 mg / mL), 40 mM human IgG Stidine buffer, 0.02% polysorbate 20, pH 5.8.

[0176] The results are shown in Table 3. The results are shown in Table 3. It was found that the formation of aggregates was small when 9% sucrose was used. It was found that the formation of aggregates was minimal when the

[0177] [Table 3]

[0178] Example 5: Buffer Screening The buffer for freeze-dried injections has a proven track record of use in biopharmaceuticals, and has a buffer content of around pH 4 to pH 7. The amino acids selected were histidine, citrate, phosphate, succinate, and glutamate, all of which have shock functions. The concentrations of these buffers were uniformly set at 25 mM. The other components were the same for the antibody-drug conjugates. Gate (1) (20 mg / mL), 10% trehalose hydrate, 0.02% polysorbate The pH was adjusted to 20 and 5.8.

[0179] The results are shown in Table 4. When histidine was used as a buffer, the formation of aggregates was the least. It was found that the number of

[0180] [Table 4]

[0181] [Example 6: pH Screening] The pH of freeze-dried injections after reconstitution was evaluated using histidine buffers selected in buffer screening. When administered intravenously, the pH range is between 4 and 7, taking into consideration biocompatibility and safety such as irritation. The other components were the same: antibody-drug conjugate (1) (20 mg / mL), 9% sucrose, 25 mM histidine buffer, 0.02% polysorbate 20 and Sample preparation was carried out according to the formulation preparation method (1).

[0182] The results are shown in Table 5. As the pH increased, the number of aggregates increased (Figure 5), and as the pH decreased, It was found that NPI increased by increasing the pH (Figure 6). It was found that the increase in β-amyloid and the increase in NPI could be suppressed in a balanced manner.

[0183] [Table 5]

[0184] [Example 7: Buffer concentration screening] The histidine buffer concentration selected as the buffer for the lyophilized injection ranged from 10 mM to 40 mM. The other components were the same as the antibody-drug conjugate (1) (20 The test solution consisted of 9% sucrose, 0.02% polysorbate 20, and pH 5.5. The preparation was carried out according to the formulation preparation method (1).

[0185] The results are shown in Table 6. When the histidine buffer concentration was 10 mM to 40 mM, Both showed similar stability.

[0186] [Table 6]

[0187] [Example 8: Surfactant screening] In the case of freeze-dried injections, surfactants act as interface protectants for the protein-concentrated layer during freezing and as agents for freeze-drying. It is expected to play a role as an interface protector against fine bubbles that are generated when the dried material is remelted. 0.02% to 0.04% polysorbate 20 (Tween 20) and polysorbate 80 (Tween 80). Common: antibody-drug conjugate (1) (20 mg / mL), 9% sucrose, 25 mM The histidine buffer was adjusted to pH 5.5. Sample preparation was carried out according to the formulation preparation method (2).

[0188] The results are shown in Tables 7 and 8 (wherein "PS" stands for polysorbate, "PS20 " stands for polysorbate 20 and "PS80" stands for polysorbate 80). Particulate count reduction using .02% to 0.04% polysorbate 20 or 80 was observed, especially when 0.02% to 0.04% polysorbate 80 was used. It was found that the number of particles could be suppressed (Figure 7).

[0189] [Table 7]

[0190] [Table 8]

[0191] Example 9: Vial Study The amber glass vials and the transparent glass vials are filled with freeze-dried injections. The quality changes when exposed to light at 1,000 lx and under white fluorescent lamps were compared. The components of the injection medium were antibody-drug conjugate (1) (20 mg / mL), 9% sucrose, The sample was prepared using 25 mM histidine buffer and 0.02% polysorbate 20 (pH 5.5). The preparation was carried out according to the formulation preparation method (1). The results are shown in Table 9. The results were higher when brown glass vials were used than when clear glass vials were used. It was found that the formation of aggregates (FIG. 8) and IoP (FIG. 9) could be suppressed.

[0192] [Table 9]

[0193] [Example 10: Setting of freeze-drying method] The important physical properties that serve as indicators for setting the freeze-drying method for injections are the drug content Freeze-drying of the aqueous solution and the porous cake formed during freeze-drying The collapse temperature (Tc) of the product is Tg' or Tc or lower. This results in a freeze-dried product with a cake-like porous structure. The drying process is generally The first drying process is to sublimate the free water in the frozen body, and the second is to sublimate the water bound to the solute (bound water). The secondary drying process is used to dry the free water below Tg'. It is appropriate to sublimate at the product temperature, but it is also possible to sublimate at a product temperature below Tc. . Antibody-drug conjugate (1) (20 mg / mL), 9% sucrose, 25 mM histidine Aqueous solution containing 0.03% polysorbate 80 and 100% ethylenediaminetetraacetate buffer and having a pH of 5.5 (hereinafter The Tg' of this solution (hereinafter referred to as "aqueous solution (1)") is -31°C and the Tc is -27°C. Fill 5.2 to 5.5 mL of the aqueous solution (1) into a glass vial with a capacity of approximately 10 mL and freeze-dry it. Drying methods were investigated.

[0194] (1) Study of primary drying process parameters The most important parameters in the primary drying process are shelf temperature and chamber vacuum. Cake shape (stability), redissolving ability, and drying time are greatly affected by the above parameters. Therefore, the shelf temperature is set to a range of -20℃ to 0℃ and the vacuum level in the storage room is set to a range of 5Pa to 15Pa. The range was investigated using an experimental design. Freeze-drying was performed with the parameters in Table 9 (annealing The shelf temperature for the product is -7°C.

[0195] [Table 10]

[0196] Figure 10 shows the results of the aqueous solution (1) when the shelf temperature and the vacuum level in the chamber are changed during the primary drying process. This is a contour map of the product temperature during primary drying under conditions of low shelf temperature and high vacuum in the oven. Conversely, when the shelf temperature is high and the vacuum level inside the warehouse is low, the product temperature becomes high. The shelf temperature ranged from -20℃ to 0℃ and the vacuum level in the storage was 5Pa. Although the product temperature in the range from 0.1 to 15 Pa was below Tc, the parameters other than the triangle area The meter caused partial collapse of the porous structure of the freeze-dried cake, causing the freeze-dried cake to shrink. It has become clear that the appearance will be in a shrink state. Figure 11 shows the results of the solution (1) when the shelf temperature and the vacuum level in the chamber were changed during the first drying process. This is a contour map of drying time. The primary drying time is long under conditions of low shelf temperature and high vacuum inside the chamber. On the other hand, when the shelf temperature is high and the vacuum level is low, the drying time is shorter. The velocity behavior was shown, and a correlation with the product temperature was confirmed. The area surrounded by the triangle in the product temperature contour map In this parameter range, a freeze-dried cake with a good shape can be obtained, but the primary drying process takes a long time. It became clear that a process was needed. From the viewpoint of appearance quality, it is desirable for the shape of the freeze-dried cake to be free of shrinkage. The results of the study revealed that the primary drying process requires at least 58 hours. Therefore, the total freeze-drying time was 11 hours for the freezing process and 1 hour for the secondary drying and shipping process. Since 2 hours are required, the total time is 81 hours. In other words, the freeze-drying process takes 3 days. Adding the start and end dates of freeze-drying to this, the manufacturing period was estimated to be 5 days. . On the other hand, freeze-drying is a process that is desirable for short periods of time in terms of production efficiency and production costs. The challenge was that the long freeze-drying process was necessary due to the high speed. To solve this problem, we developed a freeze-drying method that reduces shrinkage even when the primary drying process time is shortened. To obtain a freeze-dried cake, annealing was performed, which is expected to improve the porous structure of the freeze-dried cake. We focused on the step and investigated the annealing parameters.

[0197] (2) Annealing parameter setting The important parameter in the annealing process is the shelf temperature. This allows the product temperature of the frozen body to be adjusted. In this study, the porous structure of the freeze-dried cake was formed by the sublimation of ice crystals. The shelf temperature for annealing was optimized. The shelf temperature for annealing was set at a safe margin above the eutectic point. Since it is common to set the temperature at which the engine is cooled, the eutectic point is -2.5°C. In the aqueous solution (1), the experiment was carried out at -7℃, but the sublimation resistance of ice was increased by growing ice crystals in the frozen body. This reduces the specific surface area, which allows drying to proceed smoothly and makes it difficult for shrinkage to occur in the freeze-dried cake. A shelf temperature higher than -7°C was attempted to obtain a small porous structure. The temperature range is from -7℃ to 0.5℃, and freeze-drying is achieved by creating a porous structure with a small specific surface area. The aim of this study was to improve the appearance quality of the cake. The freeze-drying in this study was carried out with the parameters shown in Table 11. The total primary drying time was approximately 42 hours.

[0198] [Table 11]

[0199] Product temperature when annealed and freeze-dried at shelf temperatures ranging from -7°C to 0.5°C The shape of the freeze-dried cake (at annealing) is shown in Figure 12. Product temperature at annealing When the temperature was between -7.0°C and -4.6°C, the lyophilized case was Significant shrinkage was observed in the KI, and slight shrinkage was observed in the product temperature range of -4.2°C to -3.4°C. Although shrinkage was observed, shrinkage did not occur within the product temperature range of -2.6°C to -1.5°C. These results suggest that annealing at a shelf temperature close to the eutectic point is possible. It was found that the shrinkage decreased the more the temperature was increased, and that shrinkage did not occur near the eutectic point. In addition, even if the product temperature rises to -1.5℃ above the eutectic point, the solid state is maintained and the product remains in a stable state. It was found that freeze-dried cakes of normal shape were obtained. Until now, annealing has generally been performed at a shelf temperature about 5°C lower than the eutectic point. However, this study showed that annealing near the eutectic point produced freeze-dried products with better appearance. It was found that a dry cake was obtained. The shelf temperature for annealing was decided based on the results of this study. The temperature was set at -4°C to -2°C, near the melting point. In addition, this study showed that annealing near the eutectic point can improve the efficiency of the primary drying process. It was found that the time can be reduced by about 40 hours. By setting the temperature at -5℃ and the vacuum level at 7Pa, the product can be dried at a relatively low temperature. It was considered possible to reduce the drying time by about 18 hours. The drying parameters were set as follows:

[0200] (3) Secondary drying process parameter setting The secondary drying process is a process for drying the water bound to the solute (bound water). It is generally known that the degree of dryness after secondary drying has a significant effect on the storage stability of freeze-dried injections. The bound water can be reduced by drying at a higher temperature, but the antibody-drug complex The antibody portion of the conjugate is generally heat-labile, so capillary differential scanning calorimeter The thermal denaturation onset of the protein was measured using a volumetric spectrometry. The onset of thermal denaturation of jugate (1) was 52 to 53°C. The temperature was kept below 50°C and the degree of dryness (residual moisture) was evaluated. The shelf temperature for secondary drying was set in the range of 30°C to 50°C, and the degree of dryness was evaluated. Freeze drying was performed according to the parameters in Table 12.

[0201] [Table 12]

[0202] The shelf temperature for each secondary drying and the results for the residual moisture in the freeze-dried injections are shown in Table 13. It has been confirmed that the residual moisture in the propellant remains stable up to 5%, but In order to achieve a low level of moisture, the residual moisture was set to 1% or less. Although it is somewhat affected, the residual moisture content is 1% when the shelf temperature for secondary drying is in the range of 40℃ to 50℃. Based on the thermal denaturation onset and the results of this study, it was confirmed that the temperature range was 40℃ to 50℃. The shelf temperature for secondary drying was set at 45°C, which is midway between the two temperatures.

[0203] [Table 13]

[0204] (4) Summary The excipient for freeze-dried injections of biopharmaceuticals (protein drugs) is a protein freeze-storage agent. Sucrose and trehalose have protective properties and are free of the risk of the Maillard reaction during storage. Non-reducing disaccharides are the sugars of choice for pharmaceutical formulations. These sugars have a higher T value than other sugars. Because g' and Tc are low, the shelf temperature is lowered to prevent the product temperature from exceeding Tg' or Tc. This requires a long freeze-drying process and can lead to shrinkage of the freeze-dried cake. The aqueous solution (1) is no exception, so these issues In order to solve this problem, we investigated the primary drying, annealing process and primary drying process. Based on the results, the freeze-drying method for the aqueous solution (1) was set up as shown in Table 14. The drying method is characterized by the annealing parameters. It was found that freeze-dried cakes with significantly reduced shrinkage could be produced in a short period of time. .

[0205] [Table 14]

[0206] The freeze-dried injections produced by this freeze-drying method were stored at 50℃ for one month, and at 40℃ / 75℃ for one month. Storage stability at 25°C / 60%RH for three months, 25°C / 60%RH for three months, and 5°C for three months. Protein concentration, moisture, insoluble particulates, DAR, monomer / aggregate / fragment ratio, NPI No significant changes in quality were observed as a result of evaluation using indicators such as IoP, charge isomers, and pH. It was discovered that...

[0207] [Example 11: Evaluation of long-term storage stability] The freeze-dried injection preparation prepared in Example 10 was further aged at 40°C / 75% RH for 6 months. Storage stability of protein for 12 months at 25℃ / 60%RH and 18 months at 5℃ Concentration, moisture, insoluble particulates, DAR, monomer / aggregate / fragment ratio, NPI, IoP, The charge isomers and pH were used as indicators for evaluation. As a result, no significant changes in quality were observed. It was discovered that...

[0208] [Example 12: Further investigation of freeze-drying method] (1) Annealing parameter setting As a method for freeze-drying the aqueous solution (1), in order to grow ice crystals in the frozen body, the shelf temperature is set at -2. The annealing method was to hold the sample at 5°C for 120 minutes, but the sample was frozen in 120 minutes. The temperature of the upper part of the body has not risen to -2.5°C, and the growth of ice crystals is insufficient, so there is a slight freezing. It was found that shrinkage occurred in the dried cake. It was found that the temperature of the upper layer of the frozen body would reach -2.5°C if the temperature was extended, so annealing was performed. The incubation time was changed to 4 hours. By changing the incubation time to 4 hours, the ice crystals were allowed to grow sufficiently. It was found that this accelerated the water sublimation rate and reduced the primary drying time.

[0209] (2) Primary drying process parameter setting Anneal at -2.5°C for 4 hours and shelf temperature for primary drying process parameters The temperature range was from -10℃ to 15℃, and the vacuum level inside the chamber was from 4Pa to 30Pa. Freeze drying was performed according to the parameters in Table 15.

[0210] [Table 15]

[0211] Shelf temperature ranges from -10℃ to 10℃ and vacuum level ranges from 4Pa to 30Pa. The product temperature was below Tc, and the freeze-dried cake had a cake-like appearance without shrinkage or collapse. there were.

[0212] Of these, the shelf temperature is -5℃ to 5℃ and the vacuum level in the storage is 5Pa to 15Pa. In some cases, it is considered to be a particularly safe range, and further taking into account the drying time, the central conditions of this range The preferable conditions are set as follows: the shelf temperature is 0°C and the vacuum level inside the chamber is 10 Pa. It was determined.

[0213] (3) Summary Based on the above results, the freeze-drying method for the aqueous solution (1) was set as shown in Table 16.

[0214] [Table 16]

[0215] The freeze-dried injections produced by this freeze-drying method were stored at 40℃ / 75%RH for 3 months. Storage stability, protein concentration, moisture, insoluble particulates, DAR, monomers, aggregates, and fragments The ratio of β-amyloides, NPI, IoP, charge isomers, and pH were used as indices. No significant changes in quality were observed.

[0216] [Example 13: Examination of other formulations and freeze-drying conditions] (1) Preparation of the formulation Drug Substance (Antibody-drug conjugate (1) (20 mg / mL), 25 mM histidine, 9 % sucrose, pH 5.5) using a UF membrane kit (AMICON ULTRA-15, 30k The concentrate was then centrifuged at 3000 rpm and 5°C to concentrate the concentrate about three-fold. Drug dialysis cassette (dialysis membrane, Slide-A-lyzer, MWCO 20,000) The sample was then dialyzed with a buffer solution of the desired formulation at a volume 50 times larger than the volume of the sample injected into the dialysis cassette. Dialysis was performed twice at 5°C for more than 5 hours. The protein concentration after dialysis was approximately 5 mg / mL. Or, concentrate or dilute appropriately to approximately 50 mg / mL, and then add polysorbate Add 80 and mix to obtain antibody-drug conjugate (1) (5 mg / mL), 8% sucrose , 20 mM histidine buffer, and 0.04% polysorbate 80, and the pH was 4.0 (hereinafter referred to as “aqueous solution (2)”), antibody-drug conjugate ( 1) (50 mg / mL), 6% sucrose, 40 mM histidine buffer, and 0.04% Polysorbate 80, and an aqueous solution having a pH of 7.0 (hereinafter referred to as "aqueous solution (3)"). ), antibody-drug conjugate (1) (5 mg / mL), 8% trehalose hydrate, 20 mM histidine buffer, and 0.04% polysorbate 80, pH 4.0 (hereinafter referred to as "aqueous solution (4)"), and an antibody-drug conjugate (1) (50 mg / mL), 6% trehalose hydrate, 40 mM histidine buffer, and An aqueous solution containing 0.04% polysorbate 80 and having a pH of 7.0 (hereinafter referred to as "aqueous solution ( 5) was prepared. In addition, when calculated based on the amount per 20 mg of antibody-drug conjugate (1), the aqueous solution (2 ) contains 320 mg sucrose, 80 mM histidine buffer, and 1.6 mg polysorbate. The aqueous solution (3) contained 24 mg of sucrose, 16 mM histidine buffer, and The aqueous solution (4) contained 320 mg of triglyceride, 0.16 mg of polysorbate 80, and 0.16 mg of polysorbate 80. Rehalose hydrate, 80 mM histidine buffer, and 1.6 mg polysorbate 80 The aqueous solution (5) contains 24 mg of trehalose hydrate, 16 mM of histidine buffer, , and 0.16 mg of polysorbate 80. These solutions were filtered through a 0.22 μm filter. 5 mL of the filtered solution was added to brown paper. Fill the vials with the liquid, partially seal them with rubber stoppers in the case of freeze-dried preparations, and freeze them under the conditions in Table 17. After freeze-drying, the vials each had a rubber stopper fitted therein and were then tightened with a cap.

[0217] [Table 17]

[0218] (2) Stability testing The freeze-dried injections produced by this freeze-drying method are stable when stored at 40℃ / 75%RH. The protein concentration, moisture, insoluble particulates, DAR, monomer / aggregate / fragment ratio, The indices evaluated were NPI, IoP, charge isomers, and pH.

[0219] [Example 14: Production of antibody-drug conjugate (2) and formulation study] The anti-HER3 antibody (sequence number 155998) was produced by the method described in International Publication WO 2015 / 155998. A heavy chain consisting of the amino acid sequence set forth in SEQ ID NO:3 and a (an antibody comprising a light chain having a structure similar to that of the antibody of formula

[0220] [ka]

[0221] (wherein A represents the binding site to the antibody) and an anti-HER3 antibody linked via a thioether bond. -drug conjugate (hereinafter referred to as "antibody-drug conjugate (2)") The DAR of the antibody-drug conjugate (2) was 7.7.

[0222] The antibody-drug conjugate (2) was formulated in the same manner as in Examples 2 to 8. Screening (antibody-drug conjugate (2) (20 mg / mL), 9% sucrose , 25 mM histidine buffer, polysorbate 20 (0.01-0.1%), pH 4.9 As a result, antibody-drug conjugate (2) (20 mg / mL), Contains 9% sucrose, 25 mM histidine buffer, and 0.03% polysorbate 20. Therefore, the aqueous solution with a pH of 5.4 (hereinafter referred to as "aqueous solution (6)") is the preferred formulation. It was discovered that...

[0223] [Example 15: Production of antibody-drug conjugate (3) and formulation study] The anti-TROP2 antibody ( A heavy chain consisting of the amino acid sequence set forth in amino acid numbers 20 to 470 in SEQ ID NO:5; A light chain having an amino acid sequence set forth in amino acid numbers 21 to 234 in SEQ ID NO:6. The antibody is

[0224] [ka]

[0225] (wherein A represents the binding site to the antibody) and an anti-TROP2 antibody bound to the drug linker represented by the formula (I) via a thioether bond. Produce an antibody-drug conjugate (hereinafter referred to as "antibody-drug conjugate (3)") The DAR of the antibody-drug conjugate (3) was 4.3.

[0226] The antibody-drug conjugate (3) was formulated in the same manner as in Examples 2 to 8. Screening (antibody-drug conjugate (3) (20 mg / mL), sucrose (3 ~9%), histidine buffer (3~15mM), polysorbate 20 or polysorbate pH 80 (0.01–0.1%), pH 5–7, or antibody-drug conjugates (3 ) (15-40 mg / mL), 9% sucrose, 10 mM histidine buffer, 0.02% Polysorbate 80, pH 6.0) was used. As a result, the antibody-drug conjugate (3 ) (20 mg / mL), 9% sucrose, 10 mM histidine buffer, and 0.02% poly Resorbate 80, and an aqueous solution having a pH of 6.0 (hereinafter referred to as "aqueous solution (7)"). ), as well as antibody-drug conjugate (3) (20 mg / mL), 9% sucrose, 10 mM histidine buffer, and 0.03% polysorbate 80, pH 6.0 (hereinafter referred to as "aqueous solution (8)") was found to be a suitable formulation.

[0227] [Example 16: Production of antibody-drug conjugate (4) and formulation study] The anti-B7-H3 antibody ( A heavy chain consisting of the amino acid sequence set forth in amino acid numbers 20 to 471 in SEQ ID NO: 7; A light chain having an amino acid sequence set forth in amino acid numbers 21 to 233 in SEQ ID NO:8. The antibody is

[0228] [ka]

[0229] (wherein A represents the binding site to the antibody) and an anti-B7-H3 antibody bound to the drug linker via a thioether bond. Manufacture of antibody-drug conjugate (hereinafter referred to as "antibody-drug conjugate (4)") The DAR of the antibody-drug conjugate (4) was 4.6.

[0230] The antibody-drug conjugate (4) was formulated in the same manner as in Examples 2 to 8. Screening (antibody-drug conjugate (4) (20 mg / mL), 9% sucrose or 10% trehalose hydrate, 10 mM histidine buffer or 10 mM amber Acid buffer, polysorbate 20 or polysorbate 80 (0.005-0.04%) , pH 5.2-6.2, or antibody-drug conjugate (4) (20-60 mg / m L), 9% sucrose, 10 mM histidine buffer, polysorbate 20 (0.005- 0.04%), pH 5.2-6.2). As a result, the antibody-drug conjugate ( 4) (20 mg / mL), 9% sucrose, 10 mM histidine buffer, and 0.02% Polysorbate 20, and an aqueous solution having a pH of 5.9 (hereinafter referred to as "aqueous solution (9)"). ), and antibody-drug conjugate (4) (20 mg / mL), 9% sucrose , 10 mM histidine buffer, and 0.03% polysorbate 20, with a pH of 5. It was found that an aqueous solution of 9 (hereinafter referred to as "aqueous solution (10)") was a suitable formulation. Ta.

[0231] [Example 17: Production of antibody-drug conjugate (5) and formulation study] The anti-GPR20 antibody ( A heavy chain consisting of the amino acid sequence set forth in amino acid numbers 20 to 472 of SEQ ID NO:9; A light chain consisting of the amino acid sequence set forth in amino acid numbers 21 to 234 in SEQ ID NO: 10. An antibody comprising the formula

[0232] [ka]

[0233] (wherein A represents the binding site to the antibody) and an anti-GPR20 antibody bound via a thioether bond. Manufacture of antibody-drug conjugate (hereinafter referred to as "antibody-drug conjugate (5)") The DAR of the antibody-drug conjugate (5) was 7.8.

[0234] The antibody-drug conjugate (5) was formulated in the same manner as in Examples 2 to 8. Screening (antibody-drug conjugate (5) (20-60 mg / mL), 9% screen lysate, 10 mM histidine buffer, polysorbate 80 (0.01-0.1%), pH 5.0 to 6.5). As a result, the antibody-drug conjugate (5) (20 mg / m L), 9% sucrose, 10 mM histidine buffer, and 0.03% polysorbate 80 A suitable treatment is an aqueous solution containing the above-mentioned amine and having a pH of 5.4 (hereinafter referred to as "aqueous solution (11)"). It turned out to be the case.

[0235] [Example 18: Production of freeze-dried injections and stability test] The aqueous solutions (6) to (11) prepared in Examples 14 to 17 were mixed with the aqueous solutions (6) to (11) described in Examples 10 and 12. Each was freeze-dried in the same manner as in 2. to prepare a freeze-dried injection. The storage stability of these lyophilized injections at 40℃ / 75%RH for 1 month and 3 months was Qualitative analysis includes protein concentration, moisture, insoluble particulates, DAR, and monomer / aggregate / fragment ratios. The indicators evaluated were NPI, IoP, charge isomers, and pH. Outstanding quality right from the start No changes were observed. [Sequence List Free Text]

[0236] SEQ ID NO: 1: Amino acid sequence of the anti-HER2 antibody heavy chain SEQ ID NO: 2: Amino acid sequence of the anti-HER2 antibody light chain SEQ ID NO: 3: Amino acid sequence of the heavy chain of the anti-HER3 antibody SEQ ID NO: 4: Amino acid sequence of the light chain of the anti-HER3 antibody SEQ ID NO: 5: Amino acid sequence of the anti-TROP2 antibody heavy chain SEQ ID NO: 6: Amino acid sequence of the anti-TROP2 antibody light chain SEQ ID NO: 7: Amino acid sequence of the anti-B7-H3 antibody heavy chain SEQ ID NO: 8: Amino acid sequence of the anti-B7-H3 antibody light chain SEQ ID NO: 9: Amino acid sequence of the anti-GPR20 antibody heavy chain SEQ ID NO: 10: Amino acid sequence of the anti-GPR20 antibody light chain

Claims

1. (i) antibody-drug conjugates; (ii) a histidine buffer; (iii) sucrose or trehalose, and (iv) a surfactant, Including, Pharmaceutical compositions; wherein the antibody-drug conjugate has the formula [Formula 1] (In the formula, A represents the binding site with the antibody.) The antibody-drug conjugate is an antibody-drug conjugate in which a drug linker represented by the formula:

2. 2. The pharmaceutical composition of claim 1, wherein the surfactant is polysorbate 80 or polysorbate 20.

3. The pharmaceutical composition according to claim 1 or 2, wherein the antibody-drug conjugate has a pH of 5.0 to 6.0 when dissolved in water at a concentration of 20 mg / mL.

4. The pharmaceutical composition according to any one of claims 1 to 3, wherein the antibody in the antibody-drug conjugate is an anti-HER2 antibody, an anti-HER3 antibody, an anti-TROP2 antibody, an anti-B7-H3 antibody, or an anti-GPR20 antibody.

5. The pharmaceutical composition of claim 4, wherein the antibody in the antibody-drug conjugate is an anti-HER2 antibody.

6. The pharmaceutical composition according to claim 5, wherein the anti-HER2 antibody is an antibody comprising a heavy chain consisting of the amino acid sequence set forth in amino acid numbers 1 to 449 of SEQ ID NO: 1 and a light chain consisting of the amino acid sequence set forth in amino acid numbers 1 to 214 of SEQ ID NO: 2, or an antibody comprising a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO: 1 and a light chain consisting of the amino acid sequence set forth in SEQ ID NO:

2.

7. The pharmaceutical composition according to claim 5 or 6, wherein the average number of drug linkers bound per antibody in the antibody-drug conjugate is in the range of 7 to 8.

8. The pharmaceutical composition according to any one of claims 5 to 7, wherein the antibody-drug conjugate has a pH of 5.5 when dissolved in water at a concentration of 20 mg / mL.

9. The pharmaceutical composition of claim 4, wherein the antibody in the antibody-drug conjugate is an anti-HER3 antibody.

10. The pharmaceutical composition according to claim 9, wherein the anti-HER3 antibody is an antibody comprising a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO: 3 and a light chain consisting of the amino acid sequence set forth in SEQ ID NO: 4, or an antibody in which a lysine residue at the carboxyl terminus of the heavy chain of the antibody is deleted.

11. The pharmaceutical composition according to claim 9 or 10, wherein the average number of drug linkers bound per antibody in the antibody-drug conjugate is in the range of 7 to 8.

12. The pharmaceutical composition according to any one of claims 9 to 11, wherein the antibody-drug conjugate has a pH of 5.4 when dissolved in water at a concentration of 20 mg / mL.

13. The pharmaceutical composition according to claim 4, wherein the antibody in the antibody-drug conjugate is an anti-TROP2 antibody.

14. The pharmaceutical composition according to claim 13, wherein the anti-TROP2 antibody is an antibody comprising a heavy chain consisting of the amino acid sequence set forth in amino acid numbers 20 to 470 of SEQ ID NO: 5 and a light chain consisting of the amino acid sequence set forth in amino acid numbers 21 to 234 of SEQ ID NO: 6, or an antibody in which a lysine residue at the carboxyl terminus of the heavy chain of the antibody is deleted.

15. The pharmaceutical composition according to claim 13 or 14, wherein the average number of drug linkers bound per antibody in the antibody-drug conjugate is in the range of 3 to 5.

16. The pharmaceutical composition according to any one of claims 13 to 15, wherein the antibody-drug conjugate has a pH of 6.0 when dissolved in water at a concentration of 20 mg / mL.

17. The pharmaceutical composition of claim 4, wherein the antibody in the antibody-drug conjugate is an anti-B7-H3 antibody.

18. The pharmaceutical composition according to claim 17, wherein the anti-B7-H3 antibody is an antibody comprising a heavy chain consisting of the amino acid sequence set forth in amino acid numbers 20 to 471 of SEQ ID NO: 7 and a light chain consisting of the amino acid sequence set forth in amino acid numbers 21 to 233 of SEQ ID NO: 8, or an antibody in which a lysine residue at the carboxyl terminus of the heavy chain of the antibody is deleted.

19. The pharmaceutical composition according to claim 17 or 18, wherein the average number of drug linkers bound per antibody in the antibody-drug conjugate is in the range of 3 to 5.

20. The pharmaceutical composition according to any one of claims 17 to 19, wherein the antibody-drug conjugate has a pH of 5.9 when dissolved in water at a concentration of 20 mg / mL.

21. The pharmaceutical composition of claim 4, wherein the antibody in the antibody-drug conjugate is an anti-GPR20 antibody.

22. The pharmaceutical composition according to claim 21, wherein the anti-GPR20 antibody is an antibody comprising a heavy chain consisting of the amino acid sequence set forth in amino acid numbers 20 to 472 of SEQ ID NO: 9 and a light chain consisting of the amino acid sequence set forth in amino acid numbers 21 to 234 of SEQ ID NO: 10, or an antibody in which a lysine residue at the carboxyl terminus of the heavy chain of the antibody is deleted.

23. The pharmaceutical composition according to claim 21 or 22, wherein the average number of drug linkers bound per antibody in the antibody-drug conjugate is in the range of 7 to 8.

24. The pharmaceutical composition according to any one of claims 21 to 23, wherein the antibody-drug conjugate has a pH of 5.4 when dissolved in water at a concentration of 20 mg / mL.

25. The pharmaceutical composition according to any one of claims 1 to 24, which is an injectable preparation.

26. 26. The pharmaceutical composition of claim 25, which is an aqueous injectable solution.

27. The pharmaceutical composition of claim 26, wherein the concentration of the antibody-drug conjugate is 20 mg / mL.

28. 28. The pharmaceutical composition of claim 26 or 27, which is frozen.

29. 26. The pharmaceutical composition of claim 25, which is a lyophilized injection.

30. 30. The pharmaceutical composition of claim 29, contained in an amber vial.

31. 31. A pharmaceutical composition according to any one of claims 1 to 30 for use in the treatment of cancer.

32. (1) A predetermined amount of (i) antibody-drug conjugates; (ii) a histidine buffer; (iii) sucrose or trehalose, and (iv) preparing an aqueous solution comprising a surfactant; (2) If necessary, adjusting the pH of the aqueous solution to a predetermined value, and then The method for producing the pharmaceutical composition according to claim 29, comprising the step of: (3) freeze-drying the aqueous solution.

33. The method of claim 32, wherein the step of freeze-drying the aqueous solution comprises a process of annealing at a shelf temperature near the eutectic point of the aqueous solution; (wherein near the eutectic point refers to a range of temperatures from 1.5°C below the eutectic point to 1.5°C above the eutectic point).

34. The method of claim 33, wherein the time of the primary drying process is shortened compared to the case where annealing is performed at a shelf temperature 5° C. lower than the eutectic point.

35. The method according to claim 33 or 34, characterized in that a freeze-dried cake with less shrinkage is obtained as compared with a case where annealing is performed at a shelf temperature 5°C lower than the eutectic point.

36. The method of claim 32, wherein the step of freeze-drying the aqueous solution comprises an annealing process at a shelf temperature of -4 to -1°C.

37. The method according to claim 36, wherein the step of freeze-drying the aqueous solution further comprises a process of primary drying at a shelf temperature of -5 to 5°C and under a vacuum of 5 to 15 Pa.

38. The method according to claim 37, wherein the step of freeze-drying the aqueous solution further comprises a process of secondary drying at a shelf temperature of 40 to 50° C. and under a vacuum of 5 to 15 Pa.

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