Pharmaceutical composition comprising antibody-drug conjugate and use thereof

Abstract

The present invention relates to a pharmaceutical composition comprising an antibody-drug conjugate and use thereof. In particular, the present invention relates to a pharmaceutical composition comprising an antibody-drug conjugate in a buffer solution. The pharmaceutical composition of the present invention exhibits good stability.

Description

A pharmaceutical composition containing an antibody drug conjugate and uses thereof TECHNICAL FIELD

[0001] The present disclosure belongs to the field of pharmaceutical preparations, in particular to a pharmaceutical composition containing an antibody drug conjugate and uses thereof as an anticancer drug. BACKGROUND

[0002] The statements herein are provided only to complement the present disclosure and do not necessarily constitute the prior art.

[0003] Cadherin 17 (CDH17) is a cell surface marker belonging to the cadherin superfamily with a unique biological structure. Compared with the classic 5 repeated cadherin, it has 7 extracellular cadherin repeats and a very short 20 amino acid residue intracellular region, lacking a conserved intracellular domain (Berndorff et al., J Cell Biol. 1994, 125(6): 1353-1369). Although the biological function of CDH17 has not been fully explored, it has been reported that CDH17 can regulate water absorption in a Ca2+-dependent manner (Ahl et al., Biol. Med. Model. 2011, 8(18)), and CDH17 also maintains tissue integrity by interacting with integrins through extracellular tight junctions. It is mainly expressed in the human gastrointestinal (GI) tract and pancreas.

[0004] Antibody drug conjugates (ADCs) link monoclonal antibodies or antibody fragments to cytotoxins with biological activity through stable chemical linker compounds, making full use of the specificity of antibody binding to tumor cell surface antigens and the high efficiency of cytotoxins, while avoiding the defects of low efficacy of the former and excessive toxicity of the latter. This means that, compared with traditional chemotherapy drugs, antibody drug conjugates can precisely bind to tumor cells and reduce the impact on normal cells (Mullard A, (2013) Nature Reviews Drug Discovery, 12:329-332; DiJoseph JF, Armellino DC, (2004) Blood, 103:1807-1814).

[0005] Among several classes of small molecules with cytotoxicity for antibody drug conjugates, one class is camptothecin derivatives, which have anti-tumor effects by inhibiting topoisomerase I. There are reports of camptothecin derivative irinotecan (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]imidazo[1,2-b]quinoline-10,13(9H,15H)-dione) applied to antibody drug conjugates (ADC) in WO2014057687; Clinical Cancer Research (2016) 22(20):5097-5108; Cancer Sci (2016) 107:1039-1046. But there is still a need to further develop ADC drugs with better efficacy.

[0006] However, ADCs have more complex heterogeneous structures than antibodies, and thus present greater challenges for ADC formulations for therapeutic purposes. SUMMARY

[0007] The present disclosure provides a pharmaceutical composition comprising an antibody drug conjugate, wherein the antibody drug conjugate has the structure shown below:

[0008] wherein:

[0009] The concentration of the antibody drug conjugate is about 1 mg / mL to about 50 mg / mL, preferably about 10 mg / mL to about 50 mg / mL, more preferably about 20 mg / mL to about 40 mg / mL; the about optimal is 10%, for example, the concentration optimal is about 20 mg / mL, i.e. 20 mg / mL ± 2 mg / mL. Non-limiting examples include 1 mg / mL, 5 mg / mL, 10 mg / mL, 11 mg / mL, 12 mg / mL, 13 mg / mL, 14 mg / mL, 15 mg / mL, 16 mg / mL, 17 mg / mL, 18 mg / mL, 19 mg / mL, 20 mg / mL, 21 mg / mL, 22 mg / mL, 23 mg / mL, 24 mg / mL, 25 mg / mL, 26 mg / mL, 27 mg / mL, 28 mg / mL, 29 mg / mL, 30 mg / mL, 35 mg / mL, 40 mg / mL, 45 mg / mL, 50 mg / mL, and any range between these point values.

[0010] Ab is an anti-CDH17 antibody comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO: 03, SEQ ID NO: 04 and SEQ ID NO: 05, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO: 06, SEQ ID NO: 07 and SEQ ID NO: 08, respectively;

[0011] L1 and L2 are linking units;

[0012] y is an integer or a decimal number from 1 to 10, preferably an integer or a decimal number from 2 to 8, more preferably about 2, 3, 4, 5, 6, 7, 8, most preferably about 4;

[0013] In an alternative embodiment, Ab is an anti-CDH17 antibody comprising a heavy chain variable region as set forth in SEQ ID NO: 01, and a light chain variable region as set forth in SEQ ID NO: 02.

[0014] In an alternative embodiment, Ab is an anti-CDH17 antibody comprising a heavy chain as set forth in SEQ ID NO: 09, and a light chain as set forth in SEQ ID NO: 10.

[0015] In an alternative embodiment, Ab is an anti-CDH17 antibody comprising a heavy chain as set forth in SEQ ID NO: 11, and a light chain as set forth in SEQ ID NO: 10.

[0016] In an alternative embodiment, said L1 is selected from:

[0017] In an alternative embodiment, said L2 is -L a -L b -L c -L d - wherein:

[0018] L a As shown in general formula (II):

[0019] wherein: s 1 and s 2 each independently is an integer selected from 0-8;

[0020] Preferably, s 1 and s 2 are independently selected from 1, 2, 3, 4, 5 or 6; or, s 1 is an integer from 1-8, s 2 is 0, preferably, s 1is selected from 4, 5, 6, 7, or 8, s 2 is 0, or, s 2 is an integer selected from 2 to 8, s 1 is 2, preferably, s 2 is selected from 2, 3, 4, 5, or 6, s 1 is 2;

[0021] L b is a chemical bond;

[0022] L c is a tetrapeptide residue; preferably, L c is a tetrapeptide residue of glycine-glycine-phenylalanine-glycine (GGFG);

[0023] L d is -NR1(CR2R3)s 3 - wherein R1, R2, and R3 are the same or different, each independently hydrogen or alkyl, s 3 is 1 or 2;

[0024] wherein L a terminally connects to Ab, L d terminally connects to L1.

[0025] In some embodiments, the foregoing antibody-drug conjugate is of the following structure:

[0026] y is an integer or decimal number from 1 to 10, preferably an integer or decimal number from 2 to 8, more preferably about 2, 3, 4, 5, 6, 7, 8, further preferably about 4.

[0027] In alternative embodiments, the foregoing pharmaceutical composition further comprises a buffer; in alternative embodiments, the buffer is selected from one or more of a histidine buffer, an acetate buffer, a phosphate buffer, a citrate buffer, preferably a histidine-hydrochloric acid histidine buffer, an acetic acid-sodium acetate buffer, a disodium hydrogen phosphate-monosodium phosphate buffer, a citric acid-sodium citrate buffer, more preferably a histidine-hydrochloric acid histidine buffer or a citric acid-sodium citrate buffer.

[0028] In alternative embodiments, the concentration of the buffer of the foregoing pharmaceutical composition is from about 10 mM to about 50 mM, preferably from about 10 mM to about 30 mM, more preferably about 20 mM. The about is preferably ± 10%, e.g., a histidine buffer concentration of about 20 mM is 20 mM ± 2 mM.

[0029] In an alternative embodiment, the buffer of the foregoing pharmaceutical composition is about 10 mM to about 50 mM histidine buffer, preferably about 10 mM to about 30 mM histidine buffer, more preferably about 20 mM histidine buffer.

[0030] In an alternative embodiment, the buffer of the foregoing pharmaceutical composition is about 10 mM to about 50 mM histidine-hydrochloric acid histidine buffer, preferably about 10 mM to about 30 mM histidine-hydrochloric acid histidine buffer, more preferably about 20 mM histidine-hydrochloric acid histidine buffer.

[0031] In an alternative embodiment, the pH of the foregoing pharmaceutical composition is about 4.5 to 6.5, preferably the pH is about 5.0 to 6.0, more preferably the pH is about 5.5.

[0032] The "about" of this pH value is ± 0.2, e.g., about 5.0 is 5.0 ± 0.2 (i.e., pH is 4.8 to 5.2), about 5.5 is 5.5 ± 0.2 (i.e., pH is 5.3 to 5.7). Non-limiting examples include 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, and any range between these point values.

[0033] In an alternative embodiment, the foregoing pharmaceutical composition further comprises a surfactant. The surfactant can be selected from the group consisting of polysorbates, polyoxamers, Triton, sodium lauryl sulfate, sodium lauryl sulfonate, sodium octyl glycoside, lauryl-sulfobetaine, myristyl-sulfobetaine, linoleyl-sulfobetaine, stearyl-sulfobetaine, lauryl-sarcosine, myristyl-sarcosine, linoleyl-sarcosine, stearyl-sarcosine, linoleyl-betaine, myristyl-betaine, cetyl-betaine, lauryl-amidopropyl-betaine, cocamidopropyl-betaine, linoleamidopropyl-betaine, myristamidopropyl-betaine, palmitamidopropyl-betaine, isostearamidopropyl-betaine, myristamidopropyl-dimethylamine, palmitamidopropyl-dimethylamine, isostearamidopropyl-dimethylamine, sodium methyl cocoyl taurate, sodium methyl oleyl taurate, polyethylene glycol, polypropylene glycol, and copolymers of ethylene and propylene glycol, and the like.

[0034] In a preferred embodiment, the foregoing pharmaceutical composition further comprises a surfactant, and the surfactant is polysorbate 80, polysorbate 60, polysorbate 40, or polysorbate 20.

[0035] In a preferred embodiment, the foregoing pharmaceutical composition further comprises a surfactant, and the surfactant is polysorbate 80.

[0036] In alternative embodiments, the concentration of the surfactant in the foregoing pharmaceutical composition is about 0.01 mg / mL to about 1.0 mg / mL, preferably about 0.1 mg / mL to about 0.8 mg / mL, more preferably about 0.2 mg / mL to about 0.6 mg / mL, further preferably about 0.2 mg / mL.

[0037] about 10%, e.g., the concentration of polysorbate 80 is about 0.2 mg / mL, i.e., 0.2 mg / mL ± 0.02 mg / mL. Non-limiting examples include 0.02 mg / mL, 0.05 mg / mL, 0.1 mg / mL, 0.15 mg / mL, 0.2 mg / mL, 0.25 mg / mL, 0.3 mg / mL, 0.35 mg / mL, 0.4 mg / mL, 0.45 mg / mL, 0.5 mg / mL, 0.6 mg / mL, 0.7 mg / mL, 0.8 mg / mL, and any range between these point values.

[0038] In alternative embodiments, the concentration of the surfactant in the foregoing pharmaceutical composition is about 0.01 mg / mL to about 1.0 mg / mL, preferably about 0.1 mg / mL to about 0.8 mg / mL, more preferably about 0.2 mg / mL to about 0.6 mg / mL, further preferably about 0.2 mg / mL.

[0039] In alternative embodiments, the concentration of the surfactant in the foregoing pharmaceutical composition is about 0.01 mg / mL to about 1.0 mg / mL, preferably about 0.1 mg / mL to about 0.8 mg / mL, more preferably about 0.2 mg / mL to about 0.6 mg / mL, further preferably about 0.2 mg / mL.

[0040] In alternative embodiments, the foregoing pharmaceutical composition further comprises a stabilizer. Optionally, the stabilizer is selected from a sugar, a polyol, an amino acid, or a combination thereof.

[0041] In preferred embodiments, the foregoing stabilizer is selected from dextrose (D-glucose), sucrose, trehalose, lactose, mannitol, sorbitol, lysine, glycine, proline, or arginine.

[0042] In preferred embodiments, the foregoing stabilizer is selected from sucrose or sorbitol.

[0043] In alternative embodiments, the concentration of the stabilizer in the foregoing pharmaceutical composition is about 10 mg / mL to about 150 mg / mL, preferably about 40 mg / mL to about 120 mg / mL, more preferably about 40 mg / mL to about 80 mg / mL, further preferably about 80 mg / mL.

[0044] The about is ±10%, e.g., sucrose concentration of about 80 mg / mL, i.e., 80 mg / mL ± 8 mg / mL. Non-limiting examples include 25 mg / mL, 30 mg / mL, 35 mg / mL, 40 mg / mL, 45 mg / mL, 50 mg / mL, 55 mg / mL, 60 mg / mL, 65 mg / mL, 70 mg / mL, 75 mg / mL, 80 mg / mL, 85 mg / mL, 90 mg / mL, 95 mg / mL, 100 mg / mL, and any range between these point values.

[0045] In alternative embodiments, the stabilizer in the foregoing pharmaceutical composition is about 10 mg / mL to about 150 mg / mL of a sugar, preferably about 40 mg / mL to about 120 mg / mL of a sugar, more preferably about 40 mg / mL to about 80 mg / mL of a sugar, and even more preferably about 80 mg / mL of a sugar.

[0046] In alternative embodiments, the stabilizer in the foregoing pharmaceutical composition is about 10 mg / mL to about 150 mg / mL of sucrose, preferably about 40 mg / mL to about 120 mg / mL of sucrose, more preferably about 40 mg / mL to about 80 mg / mL of sucrose, and even more preferably about 80 mg / mL of sucrose.

[0047] In alternative embodiments, the drug loading y (DAR) can range from an average number of cytotoxic drugs per antibody binding site, exemplary drug loading y can be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, on average. y is a decimal or an integer. Non-limiting examples of y are about 4 (3.6-4.4).

[0048] In alternative embodiments, the foregoing pharmaceutical composition comprises:

[0049] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a surfactant; the pH of the composition is about 4.5-6.5; or

[0050] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a surfactant; the pH of the composition is about 4.5-6.5; or

[0051] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a stabilizer; the pH of the composition is about 4.5-6.5; or

[0052] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a buffer, (c) a surfactant; the composition having a pH of about 4.5-6.5; or

[0053] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a buffer, (c) a stabilizer; the composition having a pH of about 4.5-6.5; or

[0054] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a stabilizer, (c) a surfactant; the composition having a pH of about 4.5-6.5; or

[0055] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a buffer, (c) a surfactant, (d) a stabilizer; the composition having a pH of about 4.5-6.5;

[0056] In alternative embodiments, the foregoing pharmaceutical compositions comprise the following components:

[0057] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a buffer; the composition having a pH of about 4.5-6.5; or

[0058] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 0.01 mg / mL to about 1.0 mg / mL of a surfactant; the composition having a pH of about 4.5-6.5; or

[0059] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mg / mL to about 150 mg / mL of a stabilizer; the composition having a pH of about 4.5-6.5; or

[0060] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a surfactant; the composition having a pH of about 4.5-6.5; or

[0061] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a buffer, (c) about 10 mg / mL to about 150 mg / mL of a stabilizer; the composition having a pH of about 4.5-6.5; or

[0062] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mg / mL to about 150 mg / mL of a stabilizer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a surfactant; the composition has a pH of about 4.5-6.5; or

[0063] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a surfactant, (d) about 10 mg / mL to about 150 mg / mL of a stabilizer; the composition has a pH of about 4.5-6.5;

[0064] In alternative embodiments, the foregoing pharmaceutical compositions comprise the following components:

[0065] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a histidine buffer; the composition has a pH of about 4.5-6.5; or

[0066] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 0.01 mg / mL to about 1.0 mg / mL of a polysorbate; the composition has a pH of about 4.5-6.5; or

[0067] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mg / mL to about 150 mg / mL of a sugar; the composition has a pH of about 4.5-6.5; or

[0068] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a histidine buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a polysorbate; the composition has a pH of about 4.5-6.5; or

[0069] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a histidine buffer, (c) about 10 mg / mL to about 150 mg / mL of a sugar; the composition has a pH of about 4.5-6.5; or

[0070] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mg / mL to about 150 mg / mL of a sugar, (c) about 0.01 mg / mL to about 1.0 mg / mL of a polysorbate; the composition has a pH of about 4.5-6.5; or

[0071] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM histidine buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL polysorbate, (d) about 10 mg / mL to about 150 mg / mL sugar; the composition has a pH of about 4.5-6.5.

[0072] In alternative embodiments, the foregoing pharmaceutical compositions comprise the following components:

[0073] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM histidine buffer; the composition has a pH of about 5.0-6.0; or

[0074] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 0.01 mg / mL to about 1.0 mg / mL polysorbate; the composition has a pH of about 5.0-6.0; or

[0075] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 40 mg / mL to about 120 mg / mL sugar; the composition has a pH of about 5.0-6.0; or

[0076] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM histidine buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL polysorbate; the composition has a pH of about 5.0-6.0; or

[0077] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM histidine buffer, (c) about 40 mg / mL to about 120 mg / mL sugar; the composition has a pH of about 5.0-6.0; or

[0078] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 40 mg / mL to about 120 mg / mL sugar, (c) about 0.01 mg / mL to about 1.0 mg / mL polysorbate; the composition has a pH of about 5.0-6.0; or

[0079] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM histidine buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL polysorbate, (d) about 40 mg / mL to about 120 mg / mL sugar, the composition having a pH of about 5.0-6.0.

[0080] In alternative embodiments, the foregoing pharmaceutical compositions comprise the following components:

[0081] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM histidine buffer; the composition having a pH of about 5.0-6.0; or

[0082] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 0.1 mg / mL to about 0.8 mg / mL polysorbate; the composition having a pH of about 5.0-6.0; or

[0083] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 40 mg / mL to about 120 mg / mL sugar; the composition having a pH of about 5.0-6.0; or

[0084] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM histidine buffer, (c) about 0.1 mg / mL to about 0.8 mg / mL polysorbate; the composition having a pH of about 5.0-6.0; or

[0085] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM histidine buffer, (c) about 40 mg / mL to about 120 mg / mL sugar; the composition having a pH of about 5.0-6.0; or

[0086] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 40 mg / mL to about 120 mg / mL sugar, (c) about 0.1 mg / mL to about 0.8 mg / mL polysorbate; the composition having a pH of about 5.0-6.0; or

[0087] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM histidine buffer, (c) about 0.1 mg / mL to about 0.8 mg / mL polysorbate, (d) about 40 mg / mL to about 120 mg / mL sugar, the composition having a pH of about 5.0-6.0.

[0088] In an alternative embodiment, the foregoing pharmaceutical composition comprises the following components:

[0089] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine-hydrochloric acid histidine buffer; the pH of the composition is about 5.0-6.0; or

[0090] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 0.2 mg / mL to about 0.6 mg / mL of polysorbate 80; the pH of the composition is about 5.0-6.0; or

[0091] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 40 mg / mL to about 80 mg / mL of sucrose; the pH of the composition is about 5.0-6.0; or

[0092] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine-hydrochloric acid histidine buffer, (c) about 0.2 mg / mL to about 0.6 mg / mL of polysorbate 80; the pH of the composition is about 5.0-6.0; or

[0093] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine-hydrochloric acid histidine buffer, (c) about 40 mg / mL to about 80 mg / mL of sucrose; the pH of the composition is about 5.0-6.0; or

[0094] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 40 mg / mL to about 80 mg / mL of sucrose, (c) about 0.2 mg / mL to about 0.6 mg / mL of polysorbate 80; the pH of the composition is about 5.0-6.0; or

[0095] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine-hydrochloric acid histidine buffer, (c) about 0.2 mg / mL to about 0.6 mg / mL of polysorbate 80, (d) about 40 mg / mL to about 80 mg / mL of sucrose, the pH of the composition is about 5.0-6.0.

[0096] In an alternative embodiment, the foregoing pharmaceutical composition comprises the following components:

[0097] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a citrate buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a polysorbate, (d) about 10 mg / mL to about 150 mg / mL of a sugar, the composition having a pH of about 4.5-6.5.

[0098] In alternative embodiments, the foregoing pharmaceutical composition comprises the following components:

[0099] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a histidine buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a polysorbate, (d) about 10 mg / mL to about 150 mg / mL of a polyol, the composition having a pH of about 4.5-6.5.

[0100] In alternative embodiments, the foregoing pharmaceutical composition comprises the following components:

[0101] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a citrate buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a polysorbate, (d) about 10 mg / mL to about 150 mg / mL of a polyol, the composition having a pH of about 4.5-6.5.

[0102] In alternative embodiments, the foregoing pharmaceutical composition comprises the following components:

[0103] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a buffer; the composition having a pH of about 5.0-6.0; or

[0104] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 0.01 mg / mL to about 1.0 mg / mL of a surfactant; the composition having a pH of about 5.0-6.0; or

[0105] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mg / mL to about 150 mg / mL of a stabilizer; the composition having a pH of about 5.0-6.0; or

[0106] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a surfactant; the composition having a pH of about 5.0-6.0; or

[0107] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a buffer, (c) about 10 mg / mL to about 150 mg / mL of a stabilizer; the composition has a pH of about 5.0-6.0; or

[0108] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mg / mL to about 150 mg / mL of a stabilizer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a surfactant; the composition has a pH of about 5.0-6.0; or

[0109] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a surfactant, (d) about 10 mg / mL to about 150 mg / mL of a stabilizer; the composition has a pH of about 5.0-6.0.

[0110] In alternative embodiments, the foregoing pharmaceutical compositions comprise the following components:

[0111] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a histidine buffer; the composition has a pH of about 5.0-6.0; or

[0112] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 0.01 mg / mL to about 1.0 mg / mL of polysorbate; the composition has a pH of about 5.0-6.0; or

[0113] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mg / mL to about 150 mg / mL of a sugar; the composition has a pH of about 5.0-6.0; or

[0114] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a histidine buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of polysorbate; the composition has a pH of about 5.0-6.0; or

[0115] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a histidine buffer, (c) about 10 mg / mL to about 150 mg / mL of a sugar; the composition has a pH of about 5.0-6.0; or

[0116] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a histidine buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a polysorbate, (d) about 10 mg / mL to about 150 mg / mL of a sugar, the composition having a pH of about 5.0-6.0.

[0117] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a histidine buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a polysorbate, (d) about 10 mg / mL to about 150 mg / mL of a sugar, the composition having a pH of about 5.0-6.0.

[0118] In an alternative embodiment, the foregoing pharmaceutical composition comprises the following components:

[0119] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a histidine buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a polysorbate, (d) about 10 mg / mL to about 150 mg / mL of a sugar, the composition having a pH of about 5.0-6.0.

[0120] In an alternative embodiment, the foregoing pharmaceutical composition comprises the following components:

[0121] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a histidine buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a polysorbate, (d) about 10 mg / mL to about 150 mg / mL of a sugar, the composition having a pH of about 5.0-6.0.

[0122] In an alternative embodiment, the foregoing pharmaceutical composition comprises the following components:

[0123] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a histidine buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a polysorbate, (d) about 10 mg / mL to about 150 mg / mL of a sugar, the composition having a pH of about 5.0-6.0.

[0124] In an alternative embodiment, the foregoing pharmaceutical composition comprises the following components:

[0125] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a citrate buffer, (c) about 0.1 mg / mL to about 0.8 mg / mL of a polysorbate, (d) about 40 mg / mL to about 120 mg / mL of a sugar, the composition having a pH of about 5.0-6.0.

[0126] In alternative embodiments, the foregoing pharmaceutical composition comprises the following components:

[0127] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a citrate buffer, (c) about 0.1 mg / mL to about 0.8 mg / mL of a polysorbate, (d) about 40 mg / mL to about 120 mg / mL of a sugar, the composition having a pH of about 5.0-6.0.

[0128] In alternative embodiments, the foregoing pharmaceutical composition comprises the following components:

[0129] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a citrate buffer, (c) about 0.1 mg / mL to about 0.8 mg / mL of a polysorbate, (d) about 40 mg / mL to about 120 mg / mL of a sugar, the composition having a pH of about 5.0-6.0.

[0130] In alternative embodiments, the foregoing pharmaceutical composition comprises the following components:

[0131] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a citrate buffer, (c) about 0.1 mg / mL to about 0.8 mg / mL of a polysorbate, (d) about 40 mg / mL to about 120 mg / mL of a sugar, the composition having a pH of about 5.0-6.0.

[0132] In alternative embodiments, the foregoing pharmaceutical composition comprises the following components:

[0133] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a citrate buffer, (c) about 0.1 mg / mL to about 0.8 mg / mL of a polysorbate, (d) about 40 mg / mL to about 120 mg / mL of a sugar, the composition having a pH of about 5.0-6.0.

[0134] In alternative embodiments, the foregoing pharmaceutical composition comprises the following components:

[0135] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine buffer, (c) about 0.2 mg / mL to about 0.6 mg / mL of a polysorbate, (d) about 40 mg / mL to about 120 mg / mL of a sugar, the composition having a pH of about 5.0-6.0.

[0136] In alternative embodiments, the foregoing pharmaceutical composition comprises the following components:

[0137] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine buffer, (c) about 0.2 mg / mL to about 0.6 mg / mL of a polysorbate, (d) about 40 mg / mL to about 80 mg / mL of a sugar, the composition having a pH of about 5.0-6.0.

[0138] In alternative embodiments, the foregoing pharmaceutical composition comprises the following components:

[0139] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine-histidine hydrochloride buffer, (c) about 0.2 mg / mL to about 0.6 mg / mL of polysorbate 80, (d) about 40 mg / mL to about 80 mg / mL of sucrose, the composition having a pH of about 5.0-6.0.

[0140] In alternative embodiments, the foregoing pharmaceutical composition comprises the following components:

[0141] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine-histidine hydrochloride buffer, (c) about 0.2 mg / mL to about 0.6 mg / mL of polysorbate 80, (d) about 40 mg / mL to about 80 mg / mL of sorbitol, the composition having a pH of about 5.0-6.0.

[0142] In alternative embodiments, the foregoing pharmaceutical composition comprises the following components:

[0143] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a citric acid-sodium citrate buffer, (c) about 0.2 mg / mL to about 0.6 mg / mL of polysorbate 80, (d) about 40 mg / mL to about 80 mg / mL of sucrose, the composition having a pH of about 5.0-6.0.

[0144] In alternative embodiments, the foregoing pharmaceutical composition comprises the following components:

[0145] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a citric acid-sodium citrate buffer, (c) about 0.2 mg / mL to about 0.6 mg / mL of polysorbate 80, (d) about 40 mg / mL to about 80 mg / mL of sorbitol, the composition having a pH of about 5.0-6.0.

[0146] In alternative embodiments, the foregoing pharmaceutical composition comprises the following components:

[0147] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine-histidine hydrochloride buffer, (c) about 0.2 mg / mL to about 0.6 mg / mL of polysorbate 80, (d) about 40 mg / mL to about 80 mg / mL of sucrose, the composition having a pH of about 5.5.

[0148] In alternative embodiments, the foregoing pharmaceutical composition comprises the following components:

[0149] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 20 mM of a histidine-histidine hydrochloride buffer, (c) about 0.2 mg / mL to about 0.6 mg / mL of polysorbate 80, (d) about 40 mg / mL to about 80 mg / mL of sucrose, the composition having a pH of about 5.0-6.0.

[0150] In alternative embodiments, the foregoing pharmaceutical composition comprises the following components:

[0151] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine-histidine hydrochloride buffer, (c) about 0.2 mg / mL of polysorbate 80, (d) about 40 mg / mL to about 80 mg / mL of sucrose, the composition having a pH of about 5.0-6.0.

[0152] In alternative embodiments, the foregoing pharmaceutical composition comprises the following components:

[0153] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine-histidine hydrochloride buffer, (c) about 0.2 mg / mL to about 0.6 mg / mL of polysorbate 80, (d) about 80 mg / mL of sucrose, the composition having a pH of about 5.0-6.0.

[0154] In alternative embodiments, the foregoing pharmaceutical composition comprises the following components:

[0155] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 20 mM histidine-histidine hydrochloride buffer, (c) about 0.2 mg / mL polysorbate 80, (d) about 80 mg / mL sucrose, the composition having a pH of about 5.5.

[0156] In alternative embodiments, the foregoing pharmaceutical composition comprises the following components:

[0157] (a) about 20 mg / mL of the antibody drug conjugate, (b) about 20 mM histidine-histidine hydrochloride buffer, (c) about 0.2 mg / mL polysorbate 80, (d) about 80 mg / mL sucrose, the composition having a pH of about 5.5.

[0158] In alternative embodiments, the foregoing pharmaceutical composition comprises the following components:

[0159] (a) about 20 mg / mL of the antibody drug conjugate, (b) about 20 mM histidine-histidine hydrochloride buffer, (c) about 0.2 mg / mL polysorbate 80, (d) about 80 mg / mL sucrose, the composition having a pH of about 5.5.

[0160] In alternative embodiments, the foregoing pharmaceutical composition comprises the following components:

[0161] (a) about 20 mg / mL of the antibody drug conjugate, (b) about 20 mM citrate-sodium citrate buffer, (c) about 0.2 mg / mL polysorbate 80, (d) about 80 mg / mL sucrose, the composition having a pH of about 5.5.

[0162] In alternative embodiments, the foregoing pharmaceutical composition comprises the following components:

[0163] (a) about 20 mg / mL of the antibody drug conjugate, (b) about 20 mM citrate-sodium citrate buffer, (c) about 0.2 mg / mL polysorbate 80, (d) about 80 mg / mL sucrose, the composition having a pH of about 5.5. In alternative embodiments, the foregoing pharmaceutical composition of any one of the foregoing is a liquid formulation.

[0164] The present disclosure also provides a lyophilized formulation comprising an antibody drug conjugate, characterized in that the formulation, when reconstituted, forms a pharmaceutical composition as described above.

[0165] The present disclosure also provides a lyophilized formulation comprising an antibody drug conjugate, characterized in that the lyophilized formulation is obtained by lyophilizing the aforementioned pharmaceutical composition.

[0166] The present disclosure also provides a method of preparing a lyophilized formulation comprising an antibody drug conjugate, comprising the step of lyophilizing the pharmaceutical composition as described above.

[0167] In alternative embodiments, the lyophilization in the method of preparing a lyophilized formulation comprising an antibody drug conjugate comprises the steps of pre-freezing, primary drying and secondary drying, in that order. Lyophilization is performed by freezing the formulation and subsequently subliming water at a temperature suitable for primary drying. Under these conditions, the product temperature is below the eutectic point or collapse temperature of the formulation. Typically, the temperature for primary drying ranges from about -45 to 25°C (assuming that the product remains frozen during primary drying). The size and type of the formulation, the container holding the sample (e.g., a glass vial), and the volume of the liquid determine the time required for drying, which can range from a few hours to several days (e.g., 40-60 hours). The secondary drying phase can be performed at about 0-40°C, depending mainly on the type and size of the container and the type of protein employed. The secondary drying time is determined by the desired residual moisture level in the product, and typically requires at least about 5 hours. Typically, the water content of a lyophilized formulation is less than about 5%, preferably less than about 3%.

[0168] In an alternative embodiment of the present disclosure, the lyophilization procedure is as follows:

[0169] (1) pre-freeze for 1-120 minutes at -2°C to -8°C;

[0170] (2) pre-freeze for 180-300 minutes at -45°C;

[0171] (3) dry for 1-15 minutes at -45°C, 0.1 mbar;

[0172] (4) dry for 2100-2700 minutes at -10°C to -20°C, 0.1 mbar;

[0173] (5) dry for 1-15 minutes at -10°C to -20°C, 0.01 mbar;

[0174] (6) dry for 480-900 minutes at 25°C, 0.01 mbar.

[0175] In some embodiments, the lyophilized formulation is stable for at least 16 days, at least 1 month, at least 3 months, at least 6 months, at least 12 months, at least 18 months, or at least 24 months at 2-8°C. In some embodiments, the lyophilized formulation is stable for at least 7 days, at least 14 days, at least 28 days, or at least 30 days at 40°C.

[0176] The present disclosure also provides a reconstituted solution of an antibody drug conjugate, characterized in that the reconstituted solution is prepared by reconstituting a lyophilized formulation as described above.

[0177] In alternative embodiments, the foregoing reconstituted solution comprises the following components:

[0178] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a buffer; the composition has a pH of about 4.5-6.5; or

[0179] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a surfactant; the composition has a pH of about 4.5-6.5; or

[0180] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a stabilizer; the composition has a pH of about 4.5-6.5; or

[0181] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a buffer, (c) a surfactant; the composition has a pH of about 4.5-6.5; or

[0182] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a buffer, (c) a stabilizer; the composition has a pH of about 4.5-6.5; or

[0183] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a stabilizer, (c) a surfactant; the composition has a pH of about 4.5-6.5; or

[0184] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a buffer, (c) a surfactant, (d) a stabilizer; the composition has a pH of about 4.5-6.5;

[0185] In alternative embodiments, the foregoing reconstituted solution comprises the following components:

[0186] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a buffer; the composition has a pH of about 4.5-6.5; or

[0187] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 0.01 mg / mL to about 1.0 mg / mL of a surfactant; the composition has a pH of about 4.5-6.5; or

[0188] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mg / mL to about 150 mg / mL of a stabilizer; the composition having a pH of about 4.5-6.5; or

[0189] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a surfactant; the composition having a pH of about 4.5-6.5; or

[0190] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a buffer, (c) about 10 mg / mL to about 150 mg / mL of a stabilizer; the composition having a pH of about 4.5-6.5; or

[0191] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mg / mL to about 150 mg / mL of a stabilizer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a surfactant; the composition having a pH of about 4.5-6.5; or

[0192] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a surfactant, (d) about 10 mg / mL to about 150 mg / mL of a stabilizer; the composition having a pH of about 4.5-6.5;

[0193] In an alternative embodiment, the foregoing reconstituted solution comprises the following components:

[0194] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a histidine buffer; the composition having a pH of about 4.5-6.5; or

[0195] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 0.01 mg / mL to about 1.0 mg / mL of a polysorbate; the composition having a pH of about 4.5-6.5; or

[0196] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mg / mL to about 150 mg / mL of a sugar; the composition having a pH of about 4.5-6.5; or

[0197] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM histidine buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL polysorbate; the composition having a pH of about 4.5-6.5; or

[0198] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM histidine buffer, (c) about 10 mg / mL to about 150 mg / mL sugar; the composition having a pH of about 4.5-6.5; or

[0199] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mg / mL to about 150 mg / mL sugar, (c) about 0.01 mg / mL to about 1.0 mg / mL polysorbate; the composition having a pH of about 4.5-6.5; or

[0200] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM histidine buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL polysorbate, (d) about 10 mg / mL to about 150 mg / mL sugar; the composition having a pH of about 4.5-6.5;

[0201] In alternative embodiments, the foregoing reconstitution solutions comprise the following components:

[0202] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM histidine buffer; the composition having a pH of about 5.0-6.0; or

[0203] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 0.01 mg / mL to about 1.0 mg / mL polysorbate; the composition having a pH of about 5.0-6.0; or

[0204] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 40 mg / mL to about 120 mg / mL sugar; the composition having a pH of about 5.0-6.0; or

[0205] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM histidine buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL polysorbate; the composition having a pH of about 5.0-6.0; or

[0206] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM histidine buffer, (c) about 40 mg / mL to about 120 mg / mL sugar; the composition has a pH of about 5.0-6.0; or

[0207] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 40 mg / mL to about 120 mg / mL sugar, (c) about 0.01 mg / mL to about 1.0 mg / mL polysorbate; the composition has a pH of about 5.0-6.0; or

[0208] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM histidine buffer, (c) about 0.1 mg / mL to about 0.8 mg / mL polysorbate, (d) about 40 mg / mL to about 120 mg / mL sugar, the composition has a pH of about 5.0-6.0;

[0209] In alternative embodiments, the foregoing reconstituted solutions comprise the following components:

[0210] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM histidine-histidine hydrochloride buffer; the composition has a pH of about 5.0-6.0; or

[0211] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 0.2 mg / mL to about 0.6 mg / mL polysorbate 80; the composition has a pH of about 5.0-6.0; or

[0212] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 40 mg / mL to about 80 mg / mL sucrose; the composition has a pH of about 5.0-6.0; or

[0213] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM histidine-histidine hydrochloride buffer, (c) about 0.2 mg / mL to about 0.6 mg / mL polysorbate 80; the composition has a pH of about 5.0-6.0; or

[0214] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM histidine-histidine hydrochloride buffer, (c) about 40 mg / mL to about 80 mg / mL sucrose; the composition has a pH of about 5.0-6.0; or

[0215] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 40 mg / mL to about 80 mg / mL of sucrose, (c) about 0.2 mg / mL to about 0.6 mg / mL of polysorbate 80; the composition having a pH of about 5.0-6.0; or

[0216] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine-histidine hydrochloride buffer, (c) about 0.2 mg / mL to about 0.6 mg / mL of polysorbate 80, (d) about 40 mg / mL to about 80 mg / mL of sucrose, the composition having a pH of about 5.0-6.0.

[0217] In alternative embodiments, the foregoing pharmaceutical compositions comprise the following components:

[0218] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a citrate buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a polysorbate, (d) about 10 mg / mL to about 150 mg / mL of a sugar, the composition having a pH of about 4.5-6.5.

[0219] In alternative embodiments, the foregoing pharmaceutical compositions comprise the following components:

[0220] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a histidine buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a polysorbate, (d) about 10 mg / mL to about 150 mg / mL of a polyol, the composition having a pH of about 4.5-6.5.

[0221] In alternative embodiments, the foregoing pharmaceutical compositions comprise the following components:

[0222] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a citrate buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a polysorbate, (d) about 10 mg / mL to about 150 mg / mL of a polyol, the composition having a pH of about 4.5-6.5. In alternative embodiments, the foregoing reconstitution solutions comprise the following components:

[0223] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM histidine buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL polysorbate, (d) about 10 mg / mL to about 150 mg / mL sugar, the composition having a pH of about 5.0-6.0.

[0224] In alternative embodiments, the foregoing reconstituted solution comprises the following components:

[0225] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM histidine buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL polysorbate, (d) about 10 mg / mL to about 150 mg / mL sugar, the composition having a pH of about 5.0-6.0.

[0226] In alternative embodiments, the foregoing reconstituted solution comprises the following components:

[0227] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM histidine buffer; the composition having a pH of about 5.0-6.0; or

[0228] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 0.01 mg / mL to about 1.0 mg / mL surfactant; the composition having a pH of about 5.0-6.0; or

[0229] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mg / mL to about 150 mg / mL stabilizer; the composition having a pH of about 5.0-6.0; or

[0230] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM histidine buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL surfactant; the composition having a pH of about 5.0-6.0; or

[0231] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM histidine buffer, (c) about 10 mg / mL to about 150 mg / mL stabilizer; the composition having a pH of about 5.0-6.0; or

[0232] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mg / mL to about 150 mg / mL of a stabilizer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a surfactant; the composition having a pH of about 5.0-6.0; or

[0233] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a histidine buffer, (c) about 0.1 mg / mL to about 0.8 mg / mL of a polysorbate, (d) about 10 mg / mL to about 150 mg / mL of a sugar, the composition having a pH of about 5.0-6.0.

[0234] In an alternative embodiment, the foregoing reconstitution solution comprises the following components:

[0235] (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a histidine buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a polysorbate, (d) about 40 mg / mL to about 120 mg / mL of a sugar, the composition having a pH of about 5.0-6.0.

[0236] In an alternative embodiment, the foregoing reconstitution solution comprises the following components:

[0237] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine buffer, (c) about 0.1 mg / mL to about 0.8 mg / mL of a polysorbate, (d) about 40 mg / mL to about 120 mg / mL of a sugar, the composition having a pH of about 5.0-6.0.

[0238] In an alternative embodiment, the foregoing reconstitution solution comprises the following components:

[0239] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a citrate buffer, (c) about 0.1 mg / mL to about 0.8 mg / mL of a polysorbate, (d) about 40 mg / mL to about 120 mg / mL of a sugar, the composition having a pH of about 5.0-6.0.

[0240] In an alternative embodiment, the foregoing reconstitution solution comprises the following components:

[0241] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a citrate buffer, (c) about 0.1 mg / mL to about 0.8 mg / mL of a polysorbate, (d) about 40 mg / mL to about 120 mg / mL of a sugar, the composition having a pH of about 5.0-6.0.

[0242] In an alternative embodiment, the foregoing reconstituted solution comprises the following components:

[0243] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a citrate buffer, (c) about 0.1 mg / mL to about 0.8 mg / mL of a polysorbate, (d) about 40 mg / mL to about 120 mg / mL of a sugar, the composition having a pH of about 5.0-6.0.

[0244] In an alternative embodiment, the foregoing reconstituted solution comprises the following components:

[0245] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a citrate buffer, (c) about 0.1 mg / mL to about 0.8 mg / mL of a polysorbate, (d) about 40 mg / mL to about 120 mg / mL of a sugar, the composition having a pH of about 5.0-6.0.

[0246] In an alternative embodiment, the foregoing reconstituted solution comprises the following components:

[0247] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a citrate buffer, (c) about 0.1 mg / mL to about 0.8 mg / mL of a polysorbate, (d) about 40 mg / mL to about 120 mg / mL of a sugar, the composition having a pH of about 5.0-6.0.

[0248] In an alternative embodiment, the foregoing reconstituted solution comprises the following components:

[0249] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a citrate buffer, (c) about 0.1 mg / mL to about 0.8 mg / mL of a polysorbate, (d) about 40 mg / mL to about 120 mg / mL of a sugar, the composition having a pH of about 5.0-6.0.

[0250] In an alternative embodiment, the foregoing reconstituted solution comprises the following components:

[0251] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine-histidine hydrochloride buffer, (c) about 0.2 mg / mL to about 0.6 mg / mL of polysorbate 80, (d) about 40 mg / mL to about 80 mg / mL of sucrose, the composition having a pH of about 5.0-6.0.

[0252] In alternative embodiments, the foregoing reconstituted solution comprises the following components:

[0253] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine-histidine hydrochloride buffer, (c) about 0.2 mg / mL to about 0.6 mg / mL of polysorbate 80, (d) about 40 mg / mL to about 80 mg / mL of sorbitol, the composition having a pH of about 5.0-6.0.

[0254] In alternative embodiments, the foregoing reconstituted solution comprises the following components:

[0255] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a citrate-sodium citrate buffer, (c) about 0.2 mg / mL to about 0.6 mg / mL of polysorbate 80, (d) about 40 mg / mL to about 80 mg / mL of sucrose, the composition having a pH of about 5.0-6.0.

[0256] In alternative embodiments, the foregoing reconstituted solution comprises the following components:

[0257] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a citrate-sodium citrate buffer, (c) about 0.2 mg / mL to about 0.6 mg / mL of polysorbate 80, (d) about 40 mg / mL to about 80 mg / mL of sorbitol, the composition having a pH of about 5.0-6.0.

[0258] In alternative embodiments, the foregoing reconstituted solution comprises the following components:

[0259] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine-histidine hydrochloride buffer, (c) about 0.2 mg / mL to about 0.6 mg / mL of polysorbate 80, (d) about 40 mg / mL to about 80 mg / mL of sucrose, the composition having a pH of about 5.5.

[0260] In alternative embodiments, the foregoing reconstituted solution comprises the following components:

[0261] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 20 mM histidine-hydrochloric acid histidine buffer, (c) about 0.2 mg / mL to about 0.6 mg / mL polysorbate 80, (d) about 40 mg / mL to about 80 mg / mL sucrose, the composition having a pH of about 5.0-6.0.

[0262] In an alternative embodiment, the foregoing reconstituted solution comprises the following components:

[0263] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM histidine-hydrochloric acid histidine buffer, (c) about 0.2 mg / mL polysorbate 80, (d) about 40 mg / mL to about 80 mg / mL sucrose, the composition having a pH of about 5.0-6.0.

[0264] In an alternative embodiment, the foregoing reconstituted solution comprises the following components:

[0265] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM histidine-hydrochloric acid histidine buffer, (c) about 0.2 mg / mL to about 0.6 mg / mL polysorbate 80, (d) about 80 mg / mL sucrose, the composition having a pH of about 5.0-6.0.

[0266] In an alternative embodiment, the foregoing reconstituted solution comprises the following components:

[0267] (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 20 mM histidine-hydrochloric acid histidine buffer, (c) about 0.2 mg / mL polysorbate 80, (d) about 80 mg / mL sucrose, the composition having a pH of about 5.5.

[0268] In an alternative embodiment, the foregoing reconstituted solution comprises the following components:

[0269] (a) about 20 mg / mL of the antibody drug conjugate, (b) about 20 mM histidine-hydrochloric acid histidine buffer, (c) about 0.2 mg / mL polysorbate 80, (d) about 80 mg / mL sucrose, the composition having a pH of about 5.5.

[0270] In an alternative embodiment, the foregoing reconstituted solution comprises the following components:

[0271] (a) about 20 mg / mL of the antibody drug conjugate, (b) about 20 mM of a citric acid-sodium citrate buffer, (c) about 0.2 mg / mL of polysorbate 80, (d) about 80 mg / mL of sorbitol, the composition having a pH of about 5.5.

[0272] In alternative embodiments, the foregoing reconstituted solution comprises the following components:

[0273] (a) about 20 mg / mL of the antibody drug conjugate, (b) about 20 mM of a citric acid-sodium citrate buffer, (c) about 0.2 mg / mL of polysorbate 80, (d) about 80 mg / mL of sorbitol, the composition having a pH of about 5.5.

[0274] In alternative embodiments, the foregoing reconstituted solution comprises the following components:

[0275] (a) about 20 mg / mL of the antibody drug conjugate, (b) about 20 mM of a citric acid-sodium citrate buffer, (c) about 0.2 mg / mL of polysorbate 80, (d) about 80 mg / mL of sorbitol, the composition having a pH of about 5.5.

[0276] The present disclosure also provides a method of preparing the foregoing reconstituted solution, comprising the step of reconstituting the foregoing lyophilized formulation with a solution selected from, but not limited to, water for injection, normal saline, or dextrose solution.

[0277] The present disclosure also provides a kit comprising a container filled with a pharmaceutical composition, a lyophilized formulation, or a reconstituted solution as described above. In some embodiments, the container is a neutral borosilicate glass tube manufactured vial.

[0278] The present disclosure also provides the use of the foregoing pharmaceutical composition or lyophilized formulation or reconstituted solution or kit in the manufacture of a medicament for the treatment or prevention of a disease.

[0279] The present disclosure also provides a method of treating a disease, comprising providing the foregoing pharmaceutical composition or lyophilized formulation or reconstituted solution or kit.

[0280] In alternative embodiments, the disease is a cancer associated with CDH17 expression.

[0281] In alternative embodiments, the cancer is selected from gastrointestinal cancer, pancreatic cancer, gallbladder cancer, cholangiocarcinoma, gastric cancer, intestinal cancer, ovarian cancer, colorectal cancer, lung cancer, breast cancer, anal cancer, prostate cancer, kidney cancer, bladder cancer, pharynx cancer, nasal cancer, skin cancer, oral cancer, tongue cancer, esophageal cancer, vaginal cancer, cervical cancer, spleen cancer, testicular cancer, or glioblastoma.

[0282] The present disclosure also provides a pharmaceutical composition of an anti-CDH17 antibody, wherein the anti-CDH17 antibody comprises a heavy chain variable region comprising HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, respectively; and a light chain variable region comprising LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, respectively; the concentration of the anti-CDH17 antibody is about 1 mg / mL to about 100 mg / mL, preferably about 10 mg / mL to about 100 mg / mL, more preferably about 20 mg / mL to about 80 mg / mL, further preferably about 40 mg / mL, about 50 mg / mL, about 60 mg / mL, about 80 mg / mL, more further preferably about 50 mg / mL; the about is preferably 10%, such as the concentration is about 50 mg / mL, i.e. 50 mg / mL ± 5 mg / mL. Non-limiting examples include 20 mg / mL, 25 mg / mL, 30 mg / mL, 35 mg / mL, 40 mg / mL, 45 mg / mL, 50 mg / mL, 55 mg / mL, 60 mg / mL, 65 mg / mL, 70 mg / mL, 75 mg / mL, 80 mg / mL, 85 mg / mL, 90 mg / mL, 95 mg / mL, 100 mg / mL, and any range between these point values.

[0283] In an alternative embodiment, the foregoing anti-CDH17 antibody comprises a heavy chain variable region as set forth in SEQ ID NO: 01, and a light chain variable region as set forth in SEQ ID NO: 02.

[0284] In an alternative embodiment, the foregoing anti-CDH17 antibody comprises a heavy chain as set forth in SEQ ID NO: 09, and a light chain as set forth in SEQ ID NO: 10.

[0285] In an alternative embodiment, the foregoing anti-CDH17 antibody comprises a heavy chain as set forth in SEQ ID NO: 09, and a light chain as set forth in SEQ ID NO: 10.

[0286] In an alternative embodiment, the foregoing pharmaceutical composition of an anti-CDH17 antibody further comprises a histidine buffer, an acetate buffer, a phosphate buffer, preferably a histidine-histidine hydrochloride buffer, an acetate-sodium acetate buffer, a disodium hydrogen phosphate-monosodium phosphate buffer, more preferably an acetate-sodium acetate buffer.

[0287] In an alternative embodiment, the foregoing buffer is present at a concentration of about 10 mM to about 50 mM, preferably about 10 mM to about 30 mM, more preferably about 30 mM.

[0288] In an alternative embodiment, the foregoing buffer is present at a concentration of about 10 mM to about 50 mM, preferably about 10 mM to about 30 mM, more preferably about 30 mM.

[0289] In an alternative embodiment, the foregoing buffer is present at a concentration of about 10 mM to about 50 mM, preferably about 10 mM to about 30 mM, more preferably about 30 mM.

[0290] In an alternative embodiment, the foregoing pharmaceutical composition of an anti- CDH17 antibody has a pH of about 4.5 to 6.5, preferably a pH of about 4.5 to 5.5, more preferably a pH of about 5.0.

[0291] In an alternative embodiment, the foregoing pharmaceutical composition of an anti- CDH17 antibody comprises:

[0292] (a) an anti-CDH17 antibody at a concentration of 10 mg / mL to 100 mg / mL, (b) about 10 mM to about 30 mM acetate buffer; the pharmaceutical composition having a pH of about 4.5 to 6.5.

[0293] In an alternative embodiment, the foregoing pharmaceutical composition of an anti- CDH17 antibody comprises:

[0294] (a) an anti-CDH17 antibody at a concentration of about 10 mg / mL to about 100 mg / mL, (b) about 10 mM to about 30 mM acetate-sodium acetate buffer; the pharmaceutical composition having a pH of about 4.5 to 5.5.

[0295] In an alternative embodiment, the foregoing pharmaceutical composition of an anti- CDH17 antibody comprises:

[0296] (a) an anti-CDH17 antibody at a concentration of about 20 mg / mL to about 80 mg / mL, (b) about 30 mM acetate-sodium acetate buffer; the pharmaceutical composition having a pH of about 4.5 to 5.5.

[0297] In an alternative embodiment, the foregoing pharmaceutical composition of an anti- CDH17 antibody comprises:

[0298] (a) an anti-CDH17 antibody at a concentration of about 20 mg / mL to about 80 mg / mL, (b) about 30 mM acetate-sodium acetate buffer; the pharmaceutical composition having a pH of about 5.0.

[0299] In an alternative embodiment, a pharmaceutical composition of an anti-CDH17 antibody as previously described, comprising:

[0300] (a) an anti-CDH17 antibody at a concentration of about 50 mg / mL, (b) about 30 mM of an acetate-sodium acetate buffer; the pH of the pharmaceutical composition is about 5.0.

[0301] In an alternative embodiment, a pharmaceutical composition of an anti-CDH17 antibody as previously described, comprising:

[0302] (a) an anti-CDH17 antibody at a concentration of about 40 mg / mL, (b) about 30 mM of an acetate-sodium acetate buffer; the pH of the pharmaceutical composition is about 5.0.

[0303] In an alternative embodiment, a pharmaceutical composition of an anti-CDH17 antibody as previously described, comprising:

[0304] (a) an anti-CDH17 antibody at a concentration of about 60 mg / mL, (b) about 30 mM of an acetate-sodium acetate buffer; the pH of the pharmaceutical composition is about 5.0.

[0305] In an alternative embodiment, a pharmaceutical composition of an anti-CDH17 antibody as previously described, comprising:

[0306] (a) an anti-CDH17 antibody at a concentration of about 80 mg / mL, (b) about 30 mM of an acetate-sodium acetate buffer; the pH of the pharmaceutical composition is about 5.0.

[0307] As will be appreciated by those of skill in the art, one, some or all of the features of each of the embodiments described in the present disclosure can be further combined to form other embodiments of the present disclosure. The above embodiments of the present disclosure and other embodiments obtained by combination are further illustrated by the following detailed description. BRIEF DESCRIPTION OF DRAWINGS

[0308] Figure 1. MFI results for 40°C accelerated stability of F1-F4 formulations.

[0309] Figure 2. MFI results for shake stability of F1-F4 formulations.

[0310] Figure 3. MFI results for 40°C accelerated stability of AF1-AF4 formulations.

[0311] Figure 4. MFI results for shake stability of AF1-AF4 formulations.

[0312] Figure 5. MFI results for freeze-thaw stability of AF1-AF4 formulations.

[0313] DETAILED DESCRIPTION

[0314] The present disclosure provides a pharmaceutical composition that is more stable in performance and is more suitable for production and administration. Unwanted instability can include any one or more of the following: aggregation, deamidation (e.g., Asn deamidation), oxidation (e.g., Met oxidation), isomerization (e.g., Asp isomerization), clipping / hydrolysis / fragmentation (e.g., hinge region fragmentation), succinimide formation, unpaired cysteines, dissociation of toxins, etc. In particular, the pharmaceutical composition of the present disclosure comprises an antibody drug conjugate.

[0315] The term

[0316] For the purposes of the present disclosure, certain technical and scientific terms are specifically defined below. Unless specifically defined herein, all other technical and scientific terms used in the present disclosure have the meanings that are commonly understood by one of ordinary skill in the art in the field of the present disclosure.

[0317] An "antibody drug conjugate" (ADC) is an antibody linked to a cytotoxic or cell-killing small molecule drug through a linker unit.

[0318] "Drug loading" also referred to as drug-to-antibody ratio (DAR), is the average number of drugs conjugated to each antibody in an ADC. It can range, for example, from about 1 to about 10 drugs per antibody, and in certain embodiments, from about 1 to about 8 drugs per antibody, preferably from the range of 2-8, 2-7, 2-6, 2-5, 2-4, 3-4, 3-5, 5-6, 5-7, 5-8, and 6-8. Exemplary drug loading can be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 on average. The general formula of the ADCs of the present disclosure includes a collection of antibodies conjugated to drugs within the aforementioned ranges. In embodiments of the present disclosure, drug loading can be denoted as n. Drug loading can be determined by conventional methods such as UV / visible spectroscopy, mass spectrometry, ELISA assays, and HPLC.

[0319] The term "linker unit" or "linker fragment" or "linking unit" refers to a chemical structure fragment or bond that links one end to an antibody or antigen binding fragment thereof and the other end to a drug, which can be linked to other linkers before being linked to the drug.

[0320] Linkers, including extensions, spacers, and amino acid units, can be synthesized by methods known in the art, such as described in US 2005-0238649 Al. The linker can be a "cleavable linker" that facilitates release of the drug in the cell. For example, an acid-labile linker (e.g., a hydrazone), a protease-sensitive (e.g., a peptidase-sensitive) linker, a photolabile linker, a dimethyl linker, or a disulfide-containing linker (Chari et al., Cancer Research 52: 127-131 (1992); U.S. Patent No. 5,208,020) can be used.

[0321] The loading of the cytotoxic drug can be controlled by the following non-limiting methods, including:

[0322] (1) controlling the molar ratio of the linking reagent and the monoclonal antibody,

[0323] (2) controlling the reaction time and temperature,

[0324] (3) selecting different reaction reagents.

[0325] The three letter and one letter codes for amino acids used herein are as described in J. Biol. Chem, 243, p 3558 (1968).

[0326] The term "antibody" as used herein is used in the broadest sense and encompasses various antibody structures, including but not limited to, full length antibodies and antibody fragments (or antigen binding fragments, or antigen binding portions) so long as they exhibit the desired antigen-binding activity. Typically, a native, intact antibody is a four polypeptide chain structure comprising two identical heavy chains and two identical light chains connected by interchain disulfide bonds.

[0327] The engineered antibodies or antigen binding fragments of the present disclosure can be produced and purified by conventional methods. For example, the cDNA sequences encoding the heavy and light chains can be cloned and recombined into a GS expression vector. The recombinant immunoglobulin expression vector can be stably transfected into CHO cells. As a more preferred prior art, mammalian expression systems result in glycosylation of the antibody, particularly at the highly conserved N-terminal site in the Fc region. Positive clones are expanded in a bioreactor in serum-free media to produce the antibody. The antibody secreting culture fluid can be purified by conventional techniques. For example, purification can be performed using an A or G Sepharose FF column with a modified buffer. Non-specifically bound components are washed away. The bound antibody is eluted using a pH gradient method, and the antibody fraction is collected and detected by SDS-PAGE. The antibody can be filter concentrated by conventional methods. Soluble aggregates and multimers can also be removed by conventional methods, such as molecular sieving, ion exchange. The resulting product is immediately frozen, such as at -70°C, or lyophilized.

[0328] "Buffer" means a buffering agent that resists changes in pH through the action of its acid-base conjugate components. Examples of buffers that control pH in an appropriate range include acetate, succinate, gluconate, histidine, oxalate, lactate, phosphate, citrate, tartrate, fumarate, glycylglycine, and other organic acid buffers.

[0329] "Histidine buffer" is a buffer that includes a histidine ion. Examples of histidine salt buffers include histidine-hydrochloric acid, histidine-acetic acid, histidine-phosphoric acid, histidine-sulfuric acid, and the like, preferably histidine-acetic acid, and more preferably histidine-acetic acid.

[0330] "Citrate buffer" is a buffer that includes a citrate ion. Examples of citrate buffers include citric acid-sodium citrate, citric acid-potassium citrate, citric acid-calcium citrate, citric acid-magnesium citrate, and the like. Preferably, the citrate buffer is citric acid-sodium citrate.

[0331] "Succinate buffer" is a buffer that includes a succinate ion. Examples of succinate buffers include succinic acid-sodium succinate, succinic acid-potassium succinate, succinic acid-calcium succinate, and the like. Preferably, the succinate buffer is succinic acid-sodium succinate. For example, the succinic acid-sodium succinate can be prepared from succinic acid and sodium hydroxide, or from succinic acid and sodium succinate.

[0332] "Phosphate buffer" is a buffer that includes a phosphate ion. Examples of phosphate buffers include disodium hydrogen phosphate-sodium dihydrogen phosphate, disodium hydrogen phosphate-potassium dihydrogen phosphate, disodium hydrogen phosphate-citric acid, and the like. Preferably, the phosphate buffer is disodium hydrogen phosphate-sodium dihydrogen phosphate.

[0333] "Acetate buffer" is a buffer that includes an acetate ion. Examples of acetate buffers include acetic acid-sodium acetate, acetic acid-histidine, acetic acid-potassium acetate, acetic acid-calcium acetate, acetic acid-magnesium acetate, and the like. Preferably, the acetate buffer is acetic acid-sodium acetate.

[0334] "Pharmaceutical composition" means a mixture of one or more antibody drug conjugates described herein, or a physiologically / pharmaceutically acceptable salt or prodrug thereof, with other chemical components, such as physiologically / pharmaceutically acceptable carriers and excipients. The goal of a pharmaceutical composition is to facilitate administration of an antibody active ingredient to a biological entity, to promote absorption, and to facilitate delivery of the active ingredient to the appropriate biological site for action.

[0335] In the present disclosure, "pharmaceutical composition" and "formulation" are not mutually exclusive.

[0336] The solutions of the pharmaceutical compositions described in the present disclosure are, unless otherwise specified, in water as solvent.

[0337] "Lyophilized formulation" means a formulation or pharmaceutical composition obtained after a vacuum freeze-drying step of a liquid or solution formulation or pharmaceutical composition.

[0338] The terms "about," "approximately," as used herein, mean within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined (i.e., the limitations of the measurement system). For example, "about" can mean within 1 or more than 1 standard deviation, per each implementation of the disclosure. Alternatively, "about" or "approximately" can mean quantities that are within 20% of the value taught (e.g., within a range of values plus or minus the 20% of that value). Furthermore, the term "about" can mean, in the context of a biological system or process, one of skill in the art will recognize that a quantity that is "about" or "approximately" a value indicates that the quantity is within 10% of the value. Unless otherwise stated, the meaning of "about" or "approximately," where used in connection with a particular value present in the application and claims, should be assumed to be within the acceptable error range for the particular value.

[0339] The pharmaceutical compositions described in the present disclosure are capable of achieving a stable effect, wherein the antibody drug conjugate of the pharmaceutical composition substantially retains its physical stability and / or chemical stability and / or biological activity after storage, preferably, the pharmaceutical composition substantially retains its physical and chemical stability and its biological activity after storage. The storage period is generally selected based on the intended shelf life of the pharmaceutical composition. There are a variety of analytical techniques to measure protein stability, which can measure stability after storage at a selected temperature for a selected period of time.

[0340] A stable formulation is one in which no significant changes are observed: at refrigerated temperatures (2-8°C) for at least 3 months, preferably 6 months, more preferably 1 year, and even more preferably up to 2 years. In addition, a stable liquid formulation includes a liquid formulation that exhibits the desired characteristics after storage at a temperature including 25°C for a period of time including 1 month, 3 months, 6 months. A typical example of stability: no more than about 10%, preferably no more than about 5% of the antibody monomer is aggregated or degraded as measured by SEC-HPLC. By visual analysis, the formulation is a pale yellow, nearly colorless, clear liquid or colorless, or clear to slightly opalescent. The concentration, pH, and osmolality of the formulation have no more than a ±10% change. Typically, no more than about 10%, preferably no more than about 5% reduction is observed. Typically, no more than about 10%, preferably no more than about 5% aggregation is formed.

[0341] An antibody drug conjugate "retains its physical stability" in a pharmaceutical formulation if it does not show a significant increase in aggregation, precipitation and / or denaturation upon visual inspection of color and / or clarity, or as measured by UV light scattering, size exclusion chromatography (SEC) and dynamic light scattering (DLS). Changes in protein conformation can be assessed by fluorescence spectroscopy, which determines the tertiary structure of the protein, and by FTIR spectroscopy, which determines the secondary structure of the protein.

[0342] An antibody retains its chemical stability in a pharmaceutical formulation if it does not show a significant chemical change. Chemical stability can be assessed by detecting and quantifying chemically altered forms of the protein. Degradation processes that frequently alter the chemical structure of a protein include hydrolysis or truncation (assessed by methods such as size exclusion chromatography and CE-SDS), oxidation (assessed by methods such as peptide mapping in conjunction with mass spectrometry or MALDI / TOF / MS), deamidation (assessed by methods such as ion exchange chromatography, capillary isoelectric focusing, peptide mapping, isoaspartate measurement), and isomerization (assessed by measuring isoaspartate content, peptide mapping, etc.).

[0343] An antibody drug conjugate "retains its biological activity" in a pharmaceutical formulation if its biological activity at a given time is within a predetermined range of the biological activity exhibited at the time of manufacture of the pharmaceutical formulation.

[0344] "Optional" or "optionally" means that the subsequently described event or circumstance can or can not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. For example, "optionally comprising 1-3 antibody heavy chain variable regions" means that antibody heavy chain variable regions of a particular sequence can or can not be present.

[0345] "Substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1-3, of a group are independently of one another replaced with a corresponding number of substituents. It goes without saying that substituents are only in their possible chemical positions, which can or cannot be possible (experimentally or theoretically) as determined by one of skill in the art without undue effort. For example, an amino or hydroxyl group with a free hydrogen can not be stable when bound to a carbon atom with an unsaturated (e.g., olefinic) bond.

[0346] The preparation of conventional pharmaceutical compositions is described in the Chinese Pharmacopoeia.

[0347] The term "carrier" used in the context of this disclosure refers to a system that can alter the way a drug enters the body and its distribution within the body, control the rate of drug release, and deliver the drug to the target organ. Drug carrier release and targeting systems can reduce drug degradation and loss, decrease side effects, and improve bioavailability. For example, high-molecular-weight surfactants, due to their unique amphiphilic structure, can self-assemble to form various forms of aggregates, preferably such as micelles, microemulsions, gels, liquid crystals, and vesicles. These aggregates have the ability to encapsulate drug molecules while also exhibiting good membrane permeability, making them excellent drug carriers.

[0348] When applied to animals, humans, experimental subjects, cells, tissues, organs, or biological fluids, "giving" and "treatment" refer to the contact of an exogenous drug, therapeutic agent, diagnostic agent, or composition with the animal, human, subject, cell, tissue, organ, or biological fluid. "Giving" and "treatment" can refer to, for example, therapeutic, pharmacokinetic, diagnostic, research, and experimental methods. Cellular treatment includes contact between a reagent and cells, as well as contact between a reagent and a fluid, wherein the fluid is in contact with the cells. "Giving" and "treatment" also mean the treatment of, for example, cells, by means of a reagent, diagnostic agent, conjugate composition, or by means of another cell in vitro and ex vivo. When applied to humans, veterinary, or research subjects, "treatment" refers to therapeutic treatment, preventative or prophylactic measures, research, and diagnostic applications.

[0349] "Treatment" means administering an oral or topical therapeutic agent, such as a composition comprising any of the compounds disclosed herein, to a patient who has symptoms of one or more diseases, and the therapeutic agent is known to have a therapeutic effect on these symptoms. Typically, a therapeutic agent is administered in a treated patient or population in an amount that effectively relieves symptoms of one or more diseases to induce the regression of such symptoms or inhibit their progression to any clinically measurable extent. The amount of a therapeutic agent that effectively relieves any specific disease symptom (also referred to as a "therapeuticly effective amount") can vary depending on a variety of factors, such as the patient's disease state, age, and weight, and the drug's ability to produce the desired therapeutic effect in the patient. Whether the disease symptoms have been relieved can be evaluated using any clinical test that a physician or other healthcare professional typically uses to assess the severity or progression of the symptoms. Although the embodiments disclosed herein (e.g., treatment methods or products) may be ineffective in alleviating symptoms of each target disease, they should reduce symptoms of the target disease in a statistically significant number of patients, as determined by any statistical test known in the art, such as the Student t-test, chi-square test, U-test according to Mann and Whitney, Kruskal-Wallis test (H-test), Jonckheere-Terpstra test, and Wilcoxon test.

[0350] An "effective amount" includes an amount sufficient to effect an improvement or prevention of a symptom or condition of a medical disorder. An effective amount also means an amount sufficient to allow or facilitate diagnosis. An effective amount for a particular patient or veterinary subject can vary depending on factors such as the condition to be treated, the overall health status of the patient, the method route and dose of administration, and the severity of side effects. An effective amount can be the maximum dose or administration regimen that avoids significant side effects or toxic effects.

[0351] "Displacement" refers to the displacement of a solvent system that solubilizes the antibody protein, e.g., a high salt or high osmotic solvent system containing the antibody protein is displaced by a physical manipulation using the buffer system of the stable formulation, such that the antibody protein exists in the stable formulation. Physical manipulations include, but are not limited to, ultrafiltration, dialysis, or resuspension following centrifugation. DETAILED DESCRIPTION

[0352] The present disclosure is further described in connection with the following examples, which are not intended to limit the scope of the disclosure. Unless otherwise indicated, the experimental methods in the examples of the present disclosure were conducted in accordance with conventional procedures, such as those described in Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory; or Molecular Cloning: A Laboratory Manual; or as suggested by the manufacturer of the materials or products. Reagents not specifically noted for their source were purchased from commercial suppliers.

[0353] I. Preparation of Antibodies

[0354] Example 1. Generation of murine hybridomas and antibody sequences

[0355] The antibodies of the present disclosure are referenced from 20B4 in PCT / CN2024 / 084343. PCT / CN2024 / 084343 is incorporated in its entirety into the present disclosure.

[0356] Anti-CDH17 antibodies were obtained by immunizing genetically modified mice encoding human immunoglobulin heavy and kappa light chain variable regions with the recombinant protein antigen human CDH17 His tag (Sino biological, Cat# 11360-H08H) and boosting 2 times with the same antigen. When the desired immune response was reached, spleen cells were harvested from each mouse and fused by electrofusion or PEG fusion of the splenic lymphocytes and myeloma cells Sp2 / 0 (ATCC® CRL-158) to obtain hybridoma cells. CRL-158) to obtain hybridoma cells.

[0357] Hybridoma clones that specifically bind to human CDH17 protein were screened by ELISA. The binding of the hybridoma supernatants on CDH17 positive cell lines AsPC1 (ATCC, CRL-1682), GP2d (Creative bioarray, CSC-J9456) and CDH17 negative cell line SW480 (ATCC, CCL-228) was analyzed by flow cytometry. In AsPC1 cells, the internalization activity of the hybridoma supernatants was measured using indirect killing assay.

[0358] After the above screening, the fully human antibody molecule 20B4 was finally selected. The variable region sequence of the fully human antibody molecule 20B4 is as follows:

[0359] 20B4 heavy chain variable region:

[0360] 20B4 light chain variable region:

[0361] Table 1. CDR sequences obtained according to Kabat numbering convention

[0362] The above anti-CDH17 antibody can further comprise an antibody heavy chain constant region and a light chain constant region; the heavy chain constant region can be selected from human IgG1, IgG2, IgG3 and IgG4 constant regions, and the light chain constant region can be selected from human antibody kappa and lambda chain constant regions. In the present disclosure, the antibody comprises a heavy chain constant region of human IgG1 and a human kappa light chain constant region. The sequence of the antibody molecule 20B4 of the present disclosure is as follows:

[0363] 20B4 heavy chain:

[0364] 20B4 light chain:

[0365] In the present disclosure, the antibody comprises a heavy chain constant region of human IgG1 and a human kappa light chain constant region. The heavy chain constant region of human IgG1 can also be other isoforms commonly used in the art, and the sequence of the antibody molecule 20B4_1 of the present disclosure is as follows:

[0366] 20B4_1 heavy chain:

[0367] 20B4_1 light chain:

[0368] II. Construction of ADC molecules

[0369] Example 2. Preparation of anti-CDH17 antibody drug conjugate (ADC)

[0370] Expression and purification of recombinant antibodies

[0371] Heavy chain expression plasmids and light chain plasmids were co-transfected into CHO cells (ATCC, Cat # CCL-61) using ExpiFectamine 293 transfection kit (ThermoFisher, A14524) or ExpiCHO-S cells (ThermoFisher, #A29127) using ExpiFectamine CHO transfection kit (ThermoFisher, A29129). Plasmid DNA concentration was brought to 1.0 pg per ml of suspension cells, LC:HC vector ratio of 1:1 based on manufacturer’s instructions. Transfected cells were incubated at 37°C, 8% CO2 on an orbital shaker for 5 to 7 days. Conditioned media was collected and antibodies were purified on an AKTA Pure 25 machine (Cytiva) using HiTrap MabSelect SuRe columns (Cytiva, #17549112). Eluted antibodies were neutralized with Tris buffer (pH 9.0) and PBS buffer exchanged. Product concentration was measured by UV absorption and quality was determined by SDS-PAGE and HPLC.

[0372] HBMAB-81 is a reformatted mAb with CDH17 binding arm from bispecific antibody BI 905771 and human IgGl backbone (see WO 2018115231A2; SEQ ID: 116, SEQ ID: 117).

[0373] Preparation of intermediates as drugs

[0374] The following intermediate compounds were used to generate antibody-drug conjugates (ADCs) of anti-CDH17 antibodies. Compound C was prepared by the methods disclosed in a PCT patent application (see WO 2020063673, filed September 25, 2019). Compound D was prepared by the methods disclosed in a PCT patent application (see WO 2022161385, filed January 26, 2022).

[0375] Anti-CDH17 antibody (5 mg / mL in PBS solution, pH 7.4) was treated with an excess of 10 mM of tris(2-carboxyethyl)phosphine (TCEP) at 37 °C for 2 hours. A sufficient molar equivalent (6-12 eq) of drug linker (Compound C or Compound D in DMSO) was added to the reduced antibody in PBS. The sample was then rotated with Compound C at 4 °C overnight. The drug loading (DAR) of the ADC was determined using a 6230 LC / MS-TOF system (Agilent), and the loading average is summarized in Table 2.

[0376] Table 2. Coupling of monoclonal antibody to linker / payload moieties

[0377] Biological testing

[0378] Test Example 1. Inhibition of tumor cell growth by ADCs

[0379] The purpose of this experiment was to test the cell-killing effect of CDH17-ADC samples according to IC 50

[0380] ​Two methods (Cell titer glo and Cytation5) were used to test the cytotoxicity of immunoconjugates in the above cell lines. For the Cytation5 method, cells were collected in exponential growth phase and distributed in 96-well plates at a density of 5000 cells / well, with the addition of 500 ng / mL of propidium iodide (abcam, cat# ab14083) and SPY650-DNA (Cytoskeleton, Inc., cat# CYSC501) at a dilution of 1:2000. Cells were incubated overnight at 37°C, 5% CO2. Immunoconjugates were diluted in cell culture medium containing 500 ng / mL propidium iodide and 2000-fold diluted SPY650-DNA, then added to each well. The final concentration of immunoconjugates ranged from 0.003 nM to 200 nM. The plates were imaged every 12 hours for a total of 96 hours using Cytation5 (Agilent) to detect the viability of cells in each well. For the Cell titer glo method. Cells were seeded into 96-well plates at 2000 cells / well. After overnight incubation, immunoconjugates were added to each well. The final concentration of immunoconjugates in the wells ranged from 0.0001 nM to 200 nM. After 5 or 7 days of incubation, the cell viability in each well was determined by Cell Titer Glo 2.0 assay (Promega).

[0381] Curves and IC values were generated using sigmoidal dose response non-linear regression fit in GraphPad Prism 50 The data of these experiments are summarized in Tables 3-4, all of the ADCs of the present application have strong inhibitory effects on tumor cell growth.

[0382] Table 3. Comparison of in vitro cytotoxicity of antibody-drug conjugates on tumor cells

[0383] Table 4. Comparison of in vitro cytotoxicity of antibody-drug conjugates on tumor cells

[0384] *: Cytotoxicity assay using Cytation5 (96 hours). Others using Celltiter Glo assay (7 days).

[0385] Test Example 2. Tumor inhibition experiment of ADCs on CDH17-positive cancer cell subcutaneous transplantation tumor model in nude mice

[0386] AsPC1 cells (pancreatic cancer) were transplanted subcutaneously into female BALB / c nude mice. When the tumor volume reached about 150-200 mm 3The engrafted mice were randomized into groups of 5 mice each. The mice were treated with ADC (6 mg / kg) by intravenous injection, Q1Wx3. The mean tumor growth inhibition (TGI) was calculated using the following formula:

[0387] TGI = ((mean(C) - mean(Co)) - (mean(T) - mean(To)) / (mean(C) - mean(Co)) * 100%; T is the current group value, C is the control group value, To and Co represent the tumor volume at the start of the test.

[0388] The results of the study are shown in Table 5.

[0389] Table 5. Efficacy of ADC in AsPC1 tumor-bearing mice

[0390] GP2d cells (colorectal cancer) were subcutaneously engrafted into female BALB / c nude mice. When the tumor volume reached approximately 100-150 mm 3 The engrafted mice were randomized into groups of 5 mice each. The mice were treated with ADC (6 mg / kg) by intravenous injection, Q1Wx3 or single dose. The results of the study are shown in Tables 6-7.

[0391] Table 6. Efficacy of ADC in GP2d tumor-bearing mice

[0392] Table 7. Comparison of efficacy of anti-CDH17 ADCs with different DAR values in GP2d tumor-bearing mice

[0393] SNU16 cells (gastric cancer) were subcutaneously engrafted into female NU / J mice. When the tumor volume reached approximately 150-200 mm 3 The engrafted mice were randomized into groups of 5 mice each. The detailed dosing strategy is described in Table 7. ADCs conjugated with Compound C have different DAR values. In the ADC-24 3 mg / kg group, the tumors started to grow again after 17 days of treatment. There was a significant difference in tumor volume between ADC-24 and ADC-21 at lower doses.

[0394] Table 7. Comparison of efficacy of anti-CDH17 ADCs with different DAR values in SNU16 tumor-bearing mice

[0395] Test Example 3. Pharmacokinetic test of ADC

[0396] To study the pharmacokinetics of ADC, ADC-10 and ADC-11 were injected intravenously into Balb / c mice (3 mice per group) at a concentration of 5 mg / kg. After injection, mouse serum samples were collected at the indicated time points (prebleed (blank), 1 hour, 4 hours, 24 hours, 2 days, 3 days, 5 days, 7 days, 14 days, 21 days) and stored at -80°C.

[0397] Detection of total antibody in mouse serum

[0398] Human CDH17 His Tag protein (Sino biological, Cat# 11360-H08H) was coated to MSD naked standard plate (Meso Scale Discovery, Cat# L15XA-3) at a concentration of 4 pg / mL, 4°C overnight. Excess unbound protein was washed away with PBST buffer, and blocked with 3% BSA in PBS buffer for 1 hour at room temperature with shaking speed of 700 rpm. After 3 washes with PBST, standard samples, QC samples, SC samples and mouse serum samples were added to the plate, incubated for 1 hour at room temperature with shaking speed of 700 rpm. The final concentration of standard samples ranged from 0.02 ng / mL to 1000 ng / mL. The quality control (QC) samples and sample control (SC) samples were diluted to 4 different concentrations (0.06 ng / ml, 0.244 ng / ml, 3.906 ng / mL and 250 ng / mL). Mouse serum samples were also diluted to the detection range. The plate was then washed 3 times with PBST, and incubated with anti-Fc antibody conjugated with sulfo-tag for 1 hour at room temperature with shaking. Finally, 150 pL of MSD GOLD Read Buffer B was added to each well, and the plate was read by MESO QuickPlex SQ 120MM (Meso Scale Discovery). The raw data was analyzed on MSD discovery workbench software, and the AUC data of mouse serum total antibody was calculated by pK solver.

[0399] Detection of ADC in mouse serum

[0400] Human CDH17 His Tag protein (Sino biological, Cat# 11360-H08H) was coated onto MSD naked standard plates (Meso Scale Discovery, Cat# L15XA-3) at a concentration of 8 pg / mL, 4°C overnight. Excess unbound protein was washed away with PBST buffer, and the plates were blocked with 3% BSA in PBS buffer for 1 hour at room temperature with shaking at 700 rpm. After 3 washes with PBST, standard samples, QC samples, SC samples, and mouse serum samples were added to the plates and incubated for 1 hour at room temperature with shaking at 700 rpm. The standard samples were in a final concentration range of 0.02 ng / mL to 1000 ng / mL. The quality control (QC) samples and sample control (SC) samples were diluted to 4 different concentrations (0.06 ng / ml, 0.244 ng / ml, 3.906 ng / mL, and 250 ng / mL). Mouse serum samples were also diluted to the detection range. The plates were then washed 3 times with PBST and incubated with sulfo-tag conjugated anti-Compound D antibody for 1 hour at room temperature with shaking. Finally, 150 pL of MSD GOLD Read Buffer B was added to each well, and the plates were read by MESO QuickPlex SQ 120MM (Meso Scale Discovery). The raw data was analyzed on MSD discovery workbench software, and the AUC data of ADC in mouse serum was calculated by pK solver.

[0401] The total antibody here refers to the antibody from the ADC, and the total antibody and the ADC are tested at the same time when the assay is performed. The results of the study are shown in Tables 8-9. The half-life and the area under the curve AUC of ADC-10 are significantly greater than those of the control ADC-11, and ADC-10 has better plasma stability.

[0402] Table 8. In vivo pharmacokinetics (PK) of total antibody after single dose administration to mice

[0403] Table 9. In vivo pharmacokinetics (PK) of conjugated antibody after single dose administration to mice

[0404] For ADC-21, ADC-24 and ADC-34, similar administration, detection and analysis methods were used. Balb / c mice (3 mice per group) were dosed by intravenous injection at a concentration of 5 mg / kg. Serum samples were collected at 13 time points: prebleed, 1 hour, 4 hours, 24 hours, 2 days, 3 days, 5 days, 7 days, 9 days, 12 days, 14 days, 17 days and 21 days. MSD bare standard plates were coated at a concentration of 2 pg / mL for total antibody and ADC PK detection. Quality control (QC) samples and sample control (SC) samples were diluted at 3 different concentrations (0.22 ng / ml, 3.52 ng / ml and 225 ng / mL).

[0405] Sulfo-tag conjugated anti-Fc antibody and anti-payload antibody were used to detect total antibody and ADC, respectively. Tables 10-11, ADC-21 and ADC-34 have greater half-life and area under the curve AUC than ADC-24, with better plasma stability.

[0406] Table 10. In vivo pharmacokinetics (PK) of total antibody after single dose administration to mice

[0407] Table 11. In vivo pharmacokinetics (PK) of conjugated antibody after single dose administration to mice

[0408] III. Formulations

[0409] Example 3. Buffer system and pH screening of antibody 20B4, 20B4_1

[0410] 30 mM Acetate-sodium acetate pH 5.0, 30 mM Histidine-acetic acid pH 5.0, 20 mM Histidine-histidine hydrochloride pH 5.5 and 20 mM Disodium hydrogen phosphate-sodium dihydrogen phosphate pH 6.0 were selected. Formulation samples of antibody 20B4 at a concentration of 40.0 mg / mL were prepared.

[0411] Table 12. Buffer system and pH screening scheme

[0412] The stability of the formulations under high temperature (40 °C), shaking (200 rpm), repeated freeze-thaw (-35 ± 5 °C 2-8 °C) conditions was investigated by detecting the appearance of the samples, SEC, IEC and NRCE purity. F4 formulation had heavy opalescence and a large number of visible particles after freeze-thaw, so it was excluded in advance and did not proceed to the subsequent sampling points.

[0413] The results of the accelerated stability test of F1-F4 at 40°C are shown in Table 13. The experimental results show that the appearance of the F1 formulation is optimal during the stability placement process, and the SEC, IEC and NRCE purity quality tests of the F1 and F3 formulations have little difference, and are better than the F2 formulation.

[0414] The present application also prepares prescription samples F1', F2', F3', F4' of antibody 20B4_1 at a concentration of 40.0 mg / mL, and the prescriptions of F1', F2', F3', F4' are consistent with F1, F2, F3, F4 respectively. The prescription of antibody 20B4_1 shows basically consistent results with the prescription of antibody 20B4.

[0415] Table 13. Results of accelerated stability test at 40°C

[0416] Note: NT = not tested.

[0417] The results of the 40°C accelerated stability of the F1-F4 formulations and the microflow imaging particle analysis MFI are shown in Figure 1. After 4W at 40°C, the insoluble particles of all formulations increased, and the insoluble particles of the F1 formulation (30mM A50) increased the least, which is consistent with the appearance results. The results of the prescriptions F1'-F4' of antibody 20B4_1 are basically consistent with the performance of the prescriptions F1-F4.

[0418] The results of the shake stability test of the prescriptions F1-F4 are shown in Table 14. The experimental results show that after 1D of 200rpm shaking, the F4 formulation (20mM P60) produces a large number of particles, and there is no obvious difference in the quality results of SEC, IEC and NRCE between the F1-F4 formulations. The results of the prescriptions F1'-F4' of antibody 20B4_1 are basically consistent with the performance of the prescriptions F1-F4.

[0419] Table 14. Results of shake stability test

[0420] The results of the shake stability of F1-F4 and the microflow imaging particle analysis MFI are shown in Figure 2. After 1D of 200rpm shaking, the insoluble particles of all formulations increased, and the insoluble particles of the F1 formulation (30mM A50) increased the least after shaking, and the performance was the best. The results of the prescriptions F1'-F4' of antibody 20B4_1 are basically consistent with the performance of the prescriptions F1-F4.

[0421] The results of the freeze-thaw stability test of the prescriptions F1-F4 are shown in Table 15. The experimental results show that after 5 cycles of freeze-thaw, the F4 formulation (20mM P60) produces obvious opalescence and a large number of particles, and there is no obvious difference in the quality results of SEC, IEC and NRCE between the F1-F4 formulations. The freeze-thaw stability results of the F1'-F4' prescriptions of antibody 20B4_1 are basically consistent with the performance of the prescriptions F1-F4.

[0422] Table 15. Results of freeze-thaw stability test

[0423] Note: NT = not tested.

[0424] In summary, the F1 / F1' formulation (30 mM A50) has the best appearance and the least increase of sub-visible particles after the accelerated condition at 40°C with shaking, and thus is selected as the optimal pH / buffer for the subsequent screening study.

[0425] Example 4. Excipient screening of antibody 20B4, 20B4_1

[0426] The optimal formulation selected in the pH / buffer screening study was added with excipients to investigate the improvement of the appearance and quality of the naked antigen solution after freeze-thaw (-35±5°C-2-8°C). The naked excipient screening scheme is shown in Table 16.

[0427] Table 16. Buffer system and pH screening scheme

[0428] The results of the freeze-thaw stability test of the formulations F1-F3 are shown in Table 17. The results show that the addition of 0.2 mg / mL polysorbate 80 significantly improves the appearance after freeze-thaw, while the addition of 80 mg / mL sucrose does not improve the appearance after freeze-thaw. There is no difference in SEC, IEC, and NRCE for different formulations. Since sucrose does not improve the appearance and quality, and the addition of surfactants may affect the subsequent ADC coupling process, the subsequent screening study is still based on the pure buffer system.

[0429] The present application also prepared the antibody 20B4_1 concentration of 40.0 mg / mL formulation samples F1', F2', F3', the formulations are consistent with F1, F2, F3 respectively, and the antibody 20B4_1 formulation shows basically consistent results with the antibody 20B4 formulation.

[0430] Table 17. Results of freeze-thaw stability test

[0431] Example 5. Concentration screening of antibody 20B4, 20B4_1

[0432] The naked antibody concentration was screened in the preferred formulation system, and the stability of the naked antibody was investigated to achieve the highest possible naked antibody concentration under the condition of ensuring the quality of the naked antibody. The naked antibody concentration screening scheme is shown in Table 18.

[0433] Table 18. Naked antibody concentration screening scheme

[0434] The results of the accelerated stability test at 40°C of the formulations F1-F3 are shown in Table 18. During the stability test, with the increase of the concentration of the formulations, the opalescence and particles increased, and the viscosity of the solution also increased, which was prone to produce bubbles. The SEC and NRCE purities decreased at 4W. Among them, the SEC and IEC purities decreased with the increase of the concentration, and the content of the polymer increased with the increase of the concentration. The increase of the concentration of the naked antibody had no effect on the NRCE purity.

[0435] The application also prepared the formulation samples F1', F2', F3' of the antibody 20B4_1 with a concentration of 40.0 mg / mL, and the formulations were consistent with F1, F2 and F3 respectively. The formulations of the antibody 20B4_1 showed basically consistent results with the formulations of the antibody 20B4.

[0436] Table 18. Results of the accelerated stability test at 40°C

[0437] The results of the freeze-thaw stability test of the formulations F1-F3 are shown in Table 19. The experimental results show that after 5 cycles of freeze-thaw, the SEC purity tends to decrease with the increase of the concentration. There is no obvious difference in the appearance, IEC and NRCE results of each formulation. The application also prepared the formulation samples F1', F2', F3' of the antibody 20B4_1 with a concentration of 40.0 mg / mL, and the formulations were consistent with F1, F2 and F3 respectively. The formulations of the antibody 20B4_1 showed basically consistent results with the formulations of the antibody 20B4.

[0438] Table 19. Results of the freeze-thaw stability test

[0439] According to the concentration screening results, when the concentration is increased, the SEC purity is observed to decrease with the increase of the concentration in the accelerated stability test at 40°C and the freeze-thaw experiment, so the low concentration is preferred. Since the naked antibody concentration in the ADC coupling process needs to be above 50.0 mg / mL, in order to meet the requirements of the ADC coupling, the naked antibody formulation concentration is selected to be 50.0 mg / mL.

[0440] Based on the above research results, the naked antibody formulation is determined to be 50.0 mg / mL, 30 mM acetic acid-sodium acetate, pH 5.0.

[0441] Example 6. Formulation prescription screening of antibody drug conjugate

[0442] According to the clinical dosage information, the concentration of the anti-CDH17 antibody drug conjugate is selected to be 20.0 mg / mL, histidine is selected as the buffer system, the effect of the ion strength of histidine on the stability of the formulation is investigated, and sucrose and polysorbate are added in the formulation to improve the stability of the antibody drug conjugate in the process of freeze-thaw, freezing and oscillation.

[0443] ADC-36 prepared in Example 2 was used as the active ingredient, and the prescription screening scheme is shown in Table 20.

[0444] Table 20. Antibody drug conjugate prescription screening scheme

[0445] The 40℃ accelerated stability investigation results are shown in Table 21. After 4W at 40℃, the SEC purity, RCE purity and DAR value decreased slightly, and there was no obvious difference between the prescriptions. The increase in peak % of IEC-A group and the ionic strength relationship was: 10mM H55>20mM H55>30mM H55. The Tween concentration had no obvious effect on the stability of each prescription.

[0446] The antibody drug conjugate ADC-37 coupled by antibody 20B4_1 was also prepared, wherein the prescription sample numbers of ADC-37 as the active ingredient were AF1', AF2', AF3', and AF4', and the prescription composition was consistent with AF1-AF4, and the prescription stability performance was basically consistent with AF1-AF4, respectively.

[0447] Table 21. 40℃ accelerated stability test results

[0448] The 40℃ accelerated stability of prescriptions AF1-AF4, and the microflow imaging particle analysis MFI detection results are shown in Figure 3. The results show that 10mM H55 and 30mM H55 have more particle increase after 4W, and prescription AF2 performs best. The prescription samples AF1', AF2', AF3', and AF4' of the antibody drug conjugate coupled by antibody 20B4_1 have basically consistent prescription performances with AF1, AF2, AF3, and AF4, respectively.

[0449] The stability test results after shaking 3D are shown in Table 22. The SEC purity, RCE purity, IEC purity and DAR value have no change, and there is no obvious difference between the prescriptions. The increase in peak % of IEC-A group and the ionic strength relationship is: 10mM H55>20mM H55>30mM H55. The prescription samples AF1', AF2', AF3', and AF4' of the antibody drug conjugate coupled by antibody 20B4_1 have basically consistent prescription performances with AF1, AF2, AF3, and AF4, respectively.

[0450] Table 22. Shaking 3D stability test results

[0451] The results of the shake 3D stability of the formulations AF1-AF4 were detected by MFI and are shown in Figure 4. The results show that AF3 (30 mM H55) had the most increase in particles after shake 3D, AF1 (10 mM H55) had the least increase in particles, and AF2 (20 mM H55) and AF4 (20 mM H55 + PS80 0.04%) had an increase in insoluble particles within an acceptable range. The antibody drug conjugate samples AF1', AF2', AF3', and AF4' coupled from antibody 20B4_1 had performances basically consistent with AF1, AF2, AF3, and AF4, respectively.

[0452] The results of the freeze-thaw stability are shown in Table 23. There was no change in SEC purity, RCE purity, IEC purity, and DAR value, and there was no significant difference between the formulations. The increase in IEC-A group peak % was related to the ionic strength as follows: 10 mM H55 > 20 mM H55 > 30 mM H55. The antibody drug conjugate samples AF1', AF2', AF3', and AF4' coupled from antibody 20B4_1 had performances basically consistent with AF1, AF2, AF3, and AF4, respectively.

[0453] Table 23. Results of freeze-thaw stability

[0454] The results of the freeze-thaw stability of the formulations AF1-AF4 were detected by MFI and are shown in Figure 5. The results show that AF3 (30 mM H55) had the most increase in particles after freeze-thaw, AF1 (10 mM H55) had the least increase in insoluble particles after freeze-thaw, and AF2 (20 mM H55) and AF4 (20 mM H55 + PS80 0.04%) had an increase in insoluble particles within an acceptable range. The antibody drug conjugate samples AF1', AF2', AF3', and AF4' coupled from antibody 20B4_1 had performances basically consistent with AF1, AF2, AF3, and AF4, respectively.

[0455] According to the above experimental results, the lower the ionic strength, the higher the risk of IEC purity reduction during stability storage; the higher the ionic strength, the higher the risk of insoluble particles during stability storage.

[0456] Considering that high-temperature acceleration can cause changes in IEC purity, especially an increasing trend in IEC-A group peak %, the applicant determined the binding activity of the samples after 2W of 40°C accelerated storage, and the results showed that the binding activity was not reduced, and thus it was considered that the change had no effect on the binding activity.

[0457] Based on the above, 20.0 mg / mL ADC, 20 mM histidine-histidine hydrochloride, 80 mg / mL sucrose and 0.2 mg / mL polysorbate 80, pH 5.5 were determined as the final formulation.

[0458] Example 7. Formulation confirmation of antibody drug conjugate

[0459] The final formulation was determined by investigating the protein stability under 25°C accelerated, 5°C and -35°C long-term storage conditions using the toxicology batch (batch number: ADC202403) sample. ADC-36 prepared in Example 2 was used as the active ingredient.

[0460] The results of the 25°C accelerated stability investigation are shown in Table 24. There were no significant changes in SEC, IEC, NRCE and DAR values after 2M at 25°C. The free drug content increased, but was all below the limit of quantification.

[0461] Table 24. Results of the 25°C long-term stability investigation

[0462] Note: <LOD is less than the limit of quantification (LOD = 20 ppm); <LOQ is less than the limit of detection (LOQ = 40 ppm)

[0463] The results of the 5°C long-term stability investigation are shown in Table 25. The experimental results showed that there were no significant changes in SEC, IEC, NRCE, DAR values and free drug.

[0464] Table 25. Results of the 5°C long-term stability investigation

[0465] The results of the -35°C long-term stability investigation are shown in Table 26. The experimental results showed that there were no significant changes in SEC, IEC, NRCE, DAR values and free drug.

[0466] Table 26. Results of the -35°C long-term stability investigation

[0467] Based on the above research results, the ADC-36 bulk formulation (20.0 mg / mL ADC, 20 mM histidine-histidine hydrochloride, 80 mg / mL sucrose and 0.2 mg / mL polysorbate 80, pH 5.5) of the present application had no significant changes in each evaluation index during the 25°C accelerated, 5°C and -35°C long-term stability investigations, and the ADC-36 of the present application had good stability.

[0468] Meanwhile, the prescription of the prepared antibody drug conjugate ADC-37 of conjugation 20B4-1 (20.0 mg / mL ADC, 20 mM histidine-hydrochloric acid, 80 mg / mL sucrose and 0.2 mg / mL polysorbate 80, pH 5.5) is subjected to the same accelerated 25°C, 5°C and -35°C long-term stability investigation method as described above, and the ADC-37 of the present application has basically the same performance as the ADC-36, so the ADC-37 of the present application also shows good stability in the prescription.

[0469] Example 8. Lyophilization process development of antibody drug conjugate

[0470] The ADC-36 prepared in Example 2 is used as the active ingredient. A prescription of 20.0 mg / mL ADC, 20 mM histidine-hydrochloric acid, 80 mg / mL sucrose and 0.2 mg / mL polysorbate 80, pH 5.5 is prepared. 5.3 mL is filled into a 20 mL vial, half-capped, and lyophilized according to the lyophilization process parameters in the following table.

[0471] Table 27. Lyophilization process parameters

[0472] The results of the comparison of the mass before and after lyophilization are shown in Table 28. There is no obvious change in the physicochemical properties before and after lyophilization, the SEC and RCE purities do not change, the DAR value does not change, no free toxin is produced, and the activity does not change. It is shown that the lyophilization process has no obvious effect on the product quality, and the prescription has good stability.

[0473] Table 28. Comparison of quality before and after lyophilization

[0474] The lyophilization is carried out according to the process parameters in Table 27, the state of the post-powder cake is observed, and the stability of the lyophilized product is investigated simultaneously. The results of the 40°C accelerated stability test are shown in Table 28. The experimental results show that there is no change in the appearance and pH of the lyophilized product during the 4W storage at 40°C, the reconstitution time is within 5 minutes, the 40°C acceleration has no great effect on the SEC, IEC and NRCE purity, DAR value of the lyophilized sample, and the free toxin content is lower than the quantitative limit. The ADC-36 has good stability in this prescription.

[0475] Meanwhile, the prescription of the prepared antibody drug conjugate ADC-37 coupled with 20B4-1 (20.0 mg / mL ADC, 20 mM histidine-hydrochloric acid, 80 mg / mL sucrose and 0.2 mg / mL polysorbate 80, pH 5.5) is freeze-dried according to the same filling and freeze-drying process parameters as described above, and the obtained freeze-dried product shows basically the same stability as the freeze-dried product of ADC-36, so the freeze-dried product of the antibody drug conjugate ADC-37 coupled with 20B4-1 of the present application also has good stability.

[0476] Table 28. Results of 40°C accelerated stability study

[0477] Note: <LOD is less than the limit of quantification (LOD = 20 ppm)

[0478] Although the specific embodiments of the present application have been described in detail, those skilled in the art will understand that various modifications and changes can be made to the details according to all the teachings of the present disclosure, and these changes are within the scope of protection of the present application. The entire scope of the present application is given by the appended claims and any equivalents thereof.

Claims

1. A pharmaceutical composition comprising an antibody drug conjugate, characterized in that, The antibody drug conjugate has a structure as shown below: wherein: Ab is an anti-CDH17 antibody comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as set forth in SEQ ID NO: 03, SEQ ID NO: 04 and SEQ ID NO: 05, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 as set forth in SEQ ID NO: 06, SEQ ID NO: 07 and SEQ ID NO: 08, respectively; Preferably, the anti-CDH17 antibody comprises a heavy chain variable region as set forth in SEQ ID NO: 01, and a light chain variable region as set forth in SEQ ID NO: 02; More preferably, the anti-CDH17 antibody comprises a heavy chain as set forth in SEQ ID NO: 09, and a light chain as set forth in SEQ ID NO: 10, or a heavy chain as set forth in SEQ ID NO: 11 and a light chain as set forth in SEQ ID NO: 10; L1 and L2 are linking units; y is an integer or a decimal number from 1 to 10, preferably an integer or a decimal number from 2 to 8, more preferably about 2, 3, 4, 5, 6, 7, 8, further preferably about 4; The concentration of the antibody drug conjugate is about 1 mg / mL to about 50 mg / mL, preferably about 10 mg / mL to about 50 mg / mL, more preferably about 20 mg / mL to about 40 mg / mL.

2. The pharmaceutical composition according to claim 1, characterized in that, said L1is selected from the group consisting of:

3. The pharmaceutical composition according to claim 1 or 2, characterized in that, said L2is -L a -L b -L c -L d -, wherein: L a As shown in general formula (II): wherein: s 1 and s 2 each independently is an integer selected from 0-8; Preferably, s 1 and s 2 is independently selected from 1, 2, 3, 4, 5 or 6; or, s 1 is an integer from 1 to 8, s 2 is 0, preferably, s 1 is selected from 4, 5, 6, 7 or 8, s 2 is 0, or, s 2 is an integer selected from 2 to 8, s 1 is 2, preferably, s 2 is selected from 2, 3, 4, 5 or 6, s 1 is 2; L b is a chemical bond; L c is a tetrapeptide residue; preferably, L c is a tetrapeptide residue of glycine-glycine-phenylalanine-glycine (GGFG); L d -NR1(CR2R3)s 3 - wherein R1, R2and R3are the same or different, each independently hydrogen or alkyl, s 3 is 1 or 2; wherein L a the end is connected to Ab, L d the end is connected to L1.

4. The pharmaceutical composition according to any one of claims 1 to 3, characterized in that, The antibody-drug conjugate is of the following structure: y is an integer or a decimal number from 1 to 10, preferably an integer or a decimal number from 2 to 8, more preferably about 2, 3, 4, 5, 6, 7, 8, further preferably about 4; Ab is as defined in claim 1.

5. The pharmaceutical composition according to any one of claims 1 to 4, characterized in that, The pharmaceutical composition further comprises a buffer; Preferably, the buffer is selected from one or more of histidine buffer, acetate buffer, phosphate buffer, citrate buffer; More preferably, the buffer is histidine-histidine hydrochloride buffer, acetate-acetate sodium buffer, disodium hydrogen phosphate-monosodium phosphate buffer or citrate-citrate sodium buffer; Further preferably, the buffer is histidine-histidine hydrochloride buffer.

6. The pharmaceutical composition of claim 5, wherein, The concentration of the buffer is about 10 mM to about 50 mM, preferably about 10 mM to about 30 mM, more preferably about 20 mM.

7. The pharmaceutical composition according to any one of claims 1-6, characterized in that, The pH of the pharmaceutical composition is about 4.5 to 6.5, preferably the pH is about 5.0 to 6.0, more preferably the pH is about 5.

5.

8. The pharmaceutical composition according to any one of claims 1 to 7, characterized in that, The pharmaceutical composition further comprises a surfactant; Preferably, the surfactant is selected from one or more of polysorbate, polyoxyl, Triton, sodium lauryl sulfate, sodium lauryl sulfonate, sodium octyl glycoside, lauryl-sulfobetaine, myristyl-sulfobetaine, linoleyl-sulfobetaine, stearyl-sulfobetaine, lauryl-sarcosine, myristyl-sarcosine, linoleyl-sarcosine, stearyl-sarcosine, linoleyl-betaine, myristyl-betaine, cetyl-betaine, lauryl-amidopropyl-betaine, cocamidopropyl-betaine, linoleamidopropyl-betaine, myristamidopropyl-betaine, palmitamidopropyl-betaine, isostearamidopropyl-betaine, myristamidopropyl-dimethylamine, palmitamidopropyl-dimethylamine, isostearamidopropyl-dimethylamine, sodium methyl cocoyl, sodium methyl oleyl taurate, polyethylene glycol, polypropylene glycol, copolymer of ethylene and propylene glycol.

9. The pharmaceutical composition of claim 8, wherein, The surfactant is polysorbate, preferably polysorbate 80, polysorbate 20, polysorbate 40, or polysorbate 60, more preferably polysorbate 80.

10. The pharmaceutical composition according to claim 8 or 9, characterized in that, The surfactant concentration is about 0.01 mg / mL to about 1.0 mg / mL, preferably about 0.1 mg / mL to about 0.8 mg / mL, more preferably about 0.2 mg / mL to about 0.6 mg / mL, further preferably about 0.2 mg / mL.

11. The pharmaceutical composition according to any one of claims 1 to 10, characterized in that, The pharmaceutical composition further comprises a stabilizer; Preferably, the stabilizer is selected from a sugar, a polyol, an amino acid, or a combination thereof; more preferably, the stabilizer is selected from D-glucose, sucrose, trehalose, lactose, mannitol, sorbitol, lysine, glycine, proline, or arginine; further preferably, the stabilizer is selected from sucrose or sorbitol.

12. The pharmaceutical composition of claim 11, wherein, The stabilizer concentration is about 10 mg / mL to about 150 mg / mL, preferably about 40 mg / mL to about 120 mg / mL, more preferably about 40 mg / mL to about 80 mg / mL, most preferably about 80 mg / mL.

13. The pharmaceutical composition according to any one of claims 1 to 12, characterized in that, comprising: (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a buffer; the pH of the composition is about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a surfactant; the pH of the composition is about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a stabilizer; the pH of the composition is about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a buffer, (c) a surfactant; the pH of the composition is about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a buffer, (c) a stabilizer; the pH of the composition is about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a buffer, (c) a stabilizer; the pH of the composition is about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a stabilizer, (c) a surfactant; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a buffer, (c) a surfactant, (d) a stabilizer; the composition having a pH of about 4.5-6.5; Preferably, the composition comprises the following components: (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a buffer; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 0.01 mg / mL to about 1.0 mg / mL surfactant; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mg / mL to about 150 mg / mL of a stabilizer; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL surfactant; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a buffer, (c) about 10 mg / mL to about 150 mg / mL of a stabilizer; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mg / mL to about 150 mg / mL of a stabilizer, (c) about 0.01 mg / mL to about 1.0 mg / mL surfactant; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL surfactant, (d) about 10 mg / mL to about 150 mg / mL of a stabilizer; the composition having a pH of about 4.5-6.5; More preferably, the composition comprises the following components: (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM histidine buffer; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 0.01 mg / mL to about 1.0 mg / mL polysorbate; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM histidine buffer; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 0.01 mg / mL to about 1.0 mg / mL polysorbate; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM histidine buffer; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 0.01 mg / mL to about 1.0 mg / mL polysorbate; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM histidine buffer; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 0.01 mg / mL to about 1.0 mg / mL polysorbate; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mg / mL to about 150 mg / mL of a sugar; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a histidine buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a polysorbate; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a histidine buffer, (c) about 10 mg / mL to about 150 mg / mL of a sugar; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mg / mL to about 150 mg / mL of a sugar, (c) about 0.01 mg / mL to about 1.0 mg / mL of a polysorbate; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a histidine buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a polysorbate, (d) about 10 mg / mL to about 150 mg / mL of a sugar; the composition having a pH of about 4.5-6.

5.

14. The pharmaceutical composition according to any one of claims 1 to 13, characterized in that, compositions comprising: (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine buffer; the composition having a pH of about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 0.01 mg / mL to about 1.0 mg / mL of a polysorbate; the composition having a pH of about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 40 mg / mL to about 120 mg / mL of a sugar; the composition having a pH of about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a polysorbate; the composition having a pH of about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine buffer, (c) about 40 mg / mL to about 120 mg / mL of a sugar; the composition having a pH of about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a polysorbate; the composition having a pH of about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine buffer, (c) about 40 mg / mL to about 120 mg / mL of a sugar; the composition having a pH of about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 40 mg / mL to about 120 mg / mL of a sugar, (c) about 0.01 mg / mL to about 1.0 mg / mL of a polysorbate; the composition has a pH of about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine buffer, (c) about 0.1 mg / mL to about 0.8 mg / mL of a polysorbate, (d) about 40 mg / mL to about 120 mg / mL of a sugar, the composition has a pH of about 5.0-6.0; More preferably, the pharmaceutical composition comprises the following components: (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine-histidine hydrochloride buffer; the composition has a pH of about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 0.2 mg / mL to about 0.6 mg / mL of polysorbate 80; the composition has a pH of about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 40 mg / mL to about 80 mg / mL of sucrose; the composition has a pH of about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine-histidine hydrochloride buffer, (c) about 0.2 mg / mL to about 0.6 mg / mL of polysorbate 80; the composition has a pH of about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine-histidine hydrochloride buffer, (c) about 40 mg / mL to about 80 mg / mL of sucrose; the composition has a pH of about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 40 mg / mL to about 80 mg / mL of sucrose, (c) about 0.2 mg / mL to about 0.6 mg / mL of polysorbate 80; the composition has a pH of about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine-histidine hydrochloride buffer, (c) about 0.2 mg / mL to about 0.6 mg / mL of polysorbate 80, (d) about 40 mg / mL to about 80 mg / mL of sucrose, the composition has a pH of about 5.0-6.

0.

15. A lyophilized formulation comprising an antibody drug conjugate, characterized in that, The lyophilized formulation, when reconstituted, forms a pharmaceutical composition of any one of claims 1 to 14.

16. A lyophilized formulation comprising an antibody drug conjugate, characterized in that, The lyophilized formulation is obtained by lyophilizing a pharmaceutical composition of any one of claims 1 to 14.

17. A method of preparing a lyophilized formulation containing an antibody drug conjugate, characterized in that, comprising the step of lyophilizing the pharmaceutical composition of any one of claims 1 to 14.

18. A reconstituted solution comprising an antibody drug conjugate, wherein the reconstituted solution is substantially free of a residual solvent. The reconstituted solution is prepared by reconstituting the lyophilized formulation of claim 16 or 17, the reconstituted solution comprising: (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a buffer; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a surfactant; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a stabilizer; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a buffer, (c) a surfactant; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a buffer, (c) a stabilizer; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a stabilizer, (c) a surfactant; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) a buffer, (c) a surfactant, (d) a stabilizer; the composition having a pH of about 4.5-6.5; Preferably, comprising: (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a buffer; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 0.01 mg / mL to about 1.0 mg / mL of a surfactant; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mg / mL to about 150 mg / mL of a stabilizer; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a surfactant; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a buffer, (c) about 10 mg / mL to about 150 mg / mL of a stabilizer; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a surfactant; the composition having a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mg / mL to about 150 mg / mL of a stabilizer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a surfactant; the composition has a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a surfactant, (d) about 10 mg / mL to about 150 mg / mL of a stabilizer; the composition has a pH of about 4.5-6.5; More preferably, comprising the following components: (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a histidine buffer; the composition has a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 0.01 mg / mL to about 1.0 mg / mL of a polysorbate; the composition has a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mg / mL to about 150 mg / mL of a sugar; the composition has a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a histidine buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a polysorbate; the composition has a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a histidine buffer, (c) about 10 mg / mL to about 150 mg / mL of a sugar; the composition has a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mg / mL to about 150 mg / mL of a sugar, (c) about 0.01 mg / mL to about 1.0 mg / mL of a polysorbate; the composition has a pH of about 4.5-6.5; or (a) about 1 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 50 mM of a histidine buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL of a polysorbate, (d) about 10 mg / mL to about 150 mg / mL of a sugar; the composition has a pH of about 4.5-6.5; Further preferably, comprising the following components: (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine buffer; the composition has a pH of about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine buffer; the composition has a pH of about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 0.01 mg / mL to about 1.0 mg / mL polysorbate; the composition has a pH of about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 40 mg / mL to about 120 mg / mL sugar; the composition has a pH of about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM histidine buffer, (c) about 0.01 mg / mL to about 1.0 mg / mL polysorbate; the composition has a pH of about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM histidine buffer, (c) about 40 mg / mL to about 120 mg / mL sugar; the composition has a pH of about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 40 mg / mL to about 120 mg / mL sugar, (c) about 0.01 mg / mL to about 1.0 mg / mL polysorbate; the composition has a pH of about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM histidine buffer, (c) about 0.1 mg / mL to about 0.8 mg / mL polysorbate, (d) about 40 mg / mL to about 120 mg / mL sugar, the composition has a pH of about 5.0-6.0; Most preferably, the pharmaceutical composition comprises the following components: (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM histidine-histidine hydrochloride buffer; the composition has a pH of about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 0.2 mg / mL to about 0.6 mg / mL polysorbate 80; the composition has a pH of about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 40 mg / mL to about 80 mg / mL sucrose; the composition has a pH of about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM histidine-histidine hydrochloride buffer, (c) about 0.2 mg / mL to about 0.6 mg / mL polysorbate 80; the composition has a pH of about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM histidine-histidine hydrochloride buffer, (c) about 0.2 mg / mL to about 0.6 mg / mL polysorbate 80; the composition has a pH of about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine-histidine hydrochloride buffer, (c) about 40 mg / mL to about 80 mg / mL of sucrose; the pH of the composition is about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 40 mg / mL to about 80 mg / mL of sucrose, (c) about 0.2 mg / mL to about 0.6 mg / mL of polysorbate 80; the pH of the composition is about 5.0-6.0; or (a) about 10 mg / mL to about 50 mg / mL of the antibody drug conjugate, (b) about 10 mM to about 30 mM of a histidine-histidine hydrochloride buffer, (c) about 0.2 mg / mL to about 0.6 mg / mL of polysorbate 80, (d) about 40 mg / mL to about 80 mg / mL of sucrose, the pH of the composition is about 5.0-6.

0.

19. A kit characterized in that, A kit comprising a container containing a pharmaceutical composition of any one of claims 1 to 12, a lyophilized formulation comprising an antibody drug conjugate of claim 16 or 17, or a reconstituted solution comprising an antibody drug conjugate of claim 18.

20. Use of a pharmaceutical composition of any one of claims 1 to 14, a lyophilized formulation comprising an antibody drug conjugate of claim 16 or 17, a reconstituted solution comprising an antibody drug conjugate of claim 18, or a kit of claim 19, in the manufacture of a medicament for treating a disease; Preferably, the disease is a cancer associated with CDH17 expression; More preferably, the cancer is selected from the group consisting of gastrointestinal cancer, pancreatic cancer, gallbladder cancer, cholangiocarcinoma, gastric cancer, intestinal cancer, ovarian cancer, colorectal cancer, lung cancer, breast cancer, anal cancer, prostate cancer, kidney cancer, bladder cancer, pharynx cancer, nasal cancer, skin cancer, oral cancer, tongue cancer, esophageal cancer, vaginal cancer, cervical cancer, spleen cancer, testicular cancer, or glioblastoma.

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