Anti-HER3 antibody-drug conjugate compositions and pharmaceutical uses thereof

Abstract

The present invention relates to an anti-HER3 antibody-drug conjugate composition and its pharmaceutical use. Specifically, the composition of the present invention comprises an anti-HER3 antibody-drug conjugate and a buffer.

Description

[Technical Field]

[0001] The present invention relates to compositions comprising anti-HER3 antibody-drug conjugates (particularly anti-HER3 antibody-exatecan analog conjugates) and pharmaceutical uses thereof. [Background technology]

[0002] Nothing herein necessarily constitutes prior art, but rather merely provides background information relevant to the present disclosure.

[0003] HER3 (epidermal growth factor receptor 3, ErbB-3, or HER3) is a member of the epidermal growth factor receptor (EGFR) family. This family includes HER1 (erbB1, EGFR), HER2 (erbB2, NEU), HER3 (erbB3), and HER4 (erbB4). Each of these receptors contains three regions: an extracellular region, a transmembrane region, and an intracellular region. The extracellular region contains four domains, and the intracellular region contains one intracellular tyrosine kinase domain for signal transduction and a cytoplasmic tail containing a phosphorylatable tyrosine residue. Cell signaling is initiated upon ligand binding to extracellular domains I and III. Normally, these receptors mediate cell division, migration, survival, and organ development. Mutations in EGFR family members result in aberrant signaling, stimulating cell survival and leading to cancer progression. The basic principle by which the HER3 receptor is activated to produce physiological effects is similar to that of other family members, but differs in that its ligands include neuregulin 1 (NRG-1) and neuregulin 2 (NRG-2). After activation, HER3 cannot form homomultimers, but can only form heterodimers with EGFR or HER2. During heterodimer formation, the intracellular domain of HER3 exhibits relatively high tyrosine phosphorylase activity. Structural analysis has revealed that the HER3 intracellular domain contains six p85 (PI-3K subunit) binding sites. This specific structure allows HER3 to recruit six PI-3Ks to the regulatory subunit site when interacting with the p85 regulatory subunit, resulting in strong activation of the PI-3K signaling pathway. In fact, the HER3 / HER2 dimer is the most active HER dimer. EGFR is widely distributed on the surface of mammalian cells, including epithelial cells, fibroblasts, glial cells, and keratinocytes. The EGFR signaling pathway plays an important role in physiological processes such as cell growth, proliferation and differentiation.

[0004] HER3 is highly expressed in various common malignant tumors, such as breast cancer, gastric cancer, ovarian cancer, prostate cancer, bladder cancer, colorectal cancer, head and neck squamous cell carcinoma, and melanoma. Unlike the EGFR gene, which is highly expressed or overactivated due to mutation, the HER3 gene has a low mutation rate, and its high expression is mainly due to increased mRNA transcription and protein translation. It is often highly expressed together with HER2, and high HER3 expression is closely related to the development, progression, and survival of various tumors, which is of great significance for the research of antitumor drugs targeting HER3.

[0005] ADCs have a more complex heterostructure than antibodies, posing an even greater challenge to their formulation for therapeutic use. Summary of the Invention [Problem to be solved by the invention]

[0006] The present disclosure provides a pharmaceutical composition comprising an anti-HER3 antibody-drug conjugate having the structure: [ka] Among them, n is 1 to 8, preferably 3 to 5; Pc is an anti-HER3 antibody containing a heavy chain variable region and a light chain variable region, of which: the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 shown in SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 shown in SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6, respectively; The pharmaceutical composition further comprises an acetate buffer, and the pH of the pharmaceutical composition is 4.9 to 5.5.

[0007] In an alternative embodiment, the pH of the pharmaceutical composition is 5.0 to 5.5.

[0008] In an alternative embodiment, the acetate buffer is an acetic acid-sodium acetate buffer.

[0009] In an alternative embodiment, in any one of the above pharmaceutical compositions, n is 3 to 5, preferably about 4.0 (ie, 4.0±0.4), more preferably 4.0.

[0010] In an alternative embodiment, the pharmaceutical composition is any one of the above, wherein the anti-HER3 antibody comprises a heavy chain variable region set forth in SEQ ID NO: 7 and a light chain variable region set forth in SEQ ID NO: 8.

[0011] In an alternative embodiment, the pharmaceutical composition is any one of the above, wherein the anti-HER3 antibody comprises a heavy chain represented by SEQ ID NO: 9 and a light chain represented by SEQ ID NO: 10.

[0012] In an alternative embodiment, the pharmaceutical composition described above has a pH value of 4.9 to 5.5, preferably 5.0 to 5.5, including, but not limited to, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, and any range between these values, preferably about pH 5.2, more preferably 5.1 to 5.4, and most preferably 5.1 to 5.3. The "about" in this pH value is ±0.2, for example, about 5.0 is 5.0±0.2 (i.e., pH 4.8 to 5.2), and about 5.2 is 5.2±0.2 (i.e., pH 5.0 to 5.4).

[0013] In alternative embodiments, any one of the pharmaceutical compositions described above further comprises a surfactant, which in some embodiments is a polysorbate (e.g., polysorbate 20, polysorbate 80), poloxamer, Triton, sodium dodecylsulfonate, sodium laurylsulfonate, sodium octylglucoside, laurylsulfobetaine, myristylsulfobetaine, linoleic sulfobetaine, stearic sulfobetaine, laurylsarcosine, myristylsarcosine, linoleic sarcosine, stearic sarcosine, linoleic betaine, myristyl betaine, or cetyl betaine. , lauramidopropyl 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, and copolymers of ethylene and propylene glycol, etc. Preferred surfactants are polysorbates, more preferably polysorbate 80 or polysorbate 20, and most preferably polysorbate 80.

[0014] In an alternative embodiment, the pharmaceutical composition described above has a surfactant concentration of 0.01 to 1.0 mg / mL, preferably 0.05 to 0.6 mg / mL, more preferably 0.1 to 0.4 mg / mL, and most preferably about 0.2 mg / mL, and non-limiting examples include 0.01 mg / mL, 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, 0.9 mg / mL, 1.0 mg / mL, and any range between these point values. The above "about" most preferably means ±10%, for example, the concentration of polysorbate 80 is about 0.2 mg / mL, i.e., 0.2 mg / mL ±0.02 mg / mL.

[0015] In some embodiments, the concentration of the surfactant is about 0.4 mg / mL. In some embodiments, the concentration of the surfactant is about 0.2 mg / mL. In some embodiments, the concentration of the surfactant is 0.4 mg / mL. In some embodiments, the concentration of the surfactant is 0.2 mg / mL.

[0016] In an alternative embodiment, the pharmaceutical composition described above further comprises a sugar. The "sugar" of the present disclosure is a sugar of the usual composition (CHO). n and derivatives thereof, including monosaccharides, disaccharides, trisaccharides, polysaccharides, sugar alcohols, reducing sugars, non-reducing sugars, etc. The sugar may be selected from glucose, sucrose, trehalose, α,α-trehalose dihydrate, lactose, fructose, maltose, dextran, glycerin, erythritol, glycerol, arabitol, sylitol, sorbitol, mannitol, melibiose, melezitose, raffinose, manninotriose, stachyose, maltose, lactulose, maltulose, sorbitol, maltitol, lactitol, iso-maltulose, etc. Preferred sugars are non-reducing disaccharides, more preferably α,α-trehalose dihydrate or sucrose, and most preferably sucrose.

[0017] In an alternative embodiment, the pharmaceutical composition described above has a sugar concentration of 10 to 150 mg / mL, preferably 20 to 120 mg / mL, more preferably 40 to 120 mg / mL, and most preferably 40 to 80 mg / mL. In some embodiments, the sugar concentration is 48 to 72 mg / mL. In some embodiments, the sugar concentration is 54 to 66 mg / mL. Non-limiting examples include 10 mg / mL, 25 mg / mL, 30 mg / mL, 35 mg / mL, 40 mg / mL, 45 mg / mL, 48 mg / mL, 50 mg / mL, 54 mg / mL, 55 mg / mL, 60 mg / mL, 66 mg / mL, 70 mg / mL, 72 mg / mL, 80 mg / mL, 90 mg / mL, 100 mg / mL, 120 mg / mL, 150 mg / mL, and any range between these point values, for example, preferably 60 to 120 mg / mL, more preferably 60 to 90 mg / mL, with the point value being preferably about 60 mg / mL. The above "about" is most preferably ±10%, and for example, the concentration of sucrose is about 60 mg / mL, i.e., 60 mg / mL ±6 mg / mL.

[0018] In an alternative embodiment, the pharmaceutical composition described above has a buffer concentration of 1 to 50 mM, preferably 10 to 20 mM. Non-limiting examples include 1 mM, 5 mM, 8 mM, 9 mM, 10 mM, 11 mM, 12 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 25 mM, 30 mM, 40 mM, 50 mM, and any range between these values. A non-limiting working range is 5 to 25 mM, preferably 5 to 15 mM, and more preferably about 10 mM. The term "about" above most preferably refers to ±10%. For example, the concentration of the acetic acid-sodium acetate buffer is about 10 mM, i.e., 10 mM ±1 mM.

[0019] In an alternative embodiment, the concentration of the antibody-drug conjugate in any one of the above pharmaceutical compositions is 1 to 50 mg / mL, preferably 1 to 40 mg / mL, and more preferably 20 to 40 mg / mL, in terms of protein concentration. 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, 40 mg / mL, and any range between these point values. Preferably, the concentration of the antibody-drug conjugate is 10 to 30 mg / mL in terms of protein concentration, more preferably, the concentration of the antibody-drug conjugate is 15 to 25 mg / mL in terms of protein concentration, and most preferably about 20 mg / mL (18 to 22 mg / mL). Specific, non-limiting examples include 20.1 mg / mL, 20.2 mg / mL, 20.3 mg / mL, 20.4 mg / mL, 20.5 mg / mL, 20.6 mg / mL, 20.7 mg / mL, 20.8 mg / mL, 20.81 mg / mL, 20.82 mg / mL, 20.83 mg / mL, 20.84 mg / mL, 20.85 mg / mL, 20.86 mg / mL, 20.87 mg / mL, The above values ​​include 20.88 mg / mL, 20.89 mg / mL, 20.9 mg / mL, 20.9 mg / mL, 20.91 mg / mL, 20.92 mg / mL, 20.93 mg / mL, 20.94 mg / mL, 20.95 mg / mL, 20.96 mg / mL, 20.97 mg / mL, 20.98 mg / mL, 20.99 mg / mL, 21 mg / mL, and any range between these values. The above protein concentration conversion means that the calculation is based on the concentration of the antibody moiety in the antibody-drug conjugate. The above "about" is most preferably 10%, and for example, a protein concentration of about 20 mg / mL, i.e., 20 mg / mL ± 2 mg / mL, is most preferred.

[0020] In some embodiments, the concentration of the anti-HER3 antibody is about 40 mg / mL. In some embodiments, the concentration of the anti-HER3 antibody is about 20 mg / mL. In some embodiments, the concentration of the anti-HER3 antibody is 40 mg / mL. In some embodiments, the concentration of the anti-HER3 antibody is 20 mg / mL.

[0021] In alternative embodiments, the range of drug loading (n) may be the average number of cytotoxic drug molecules conjugated to each anti-HER3 antibody. Non-limiting examples include about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, and any range between these values. Preferably, the range is selected from 1-8, 2-8, 2-7, 2-6, 2-5, 2-4, 3-4, 3-5, 3.5-4.7, 3.6-4.4, 5-6, 5-7, 5-8, and 6-8. Exemplary drug loading (n) may be the average of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. n is a decimal or an integer. In a non-limiting example, n is about 4 (3.6-4.4).

[0022] In an alternative embodiment, the pharmaceutical composition of any one of the above is The pharmaceutical composition contains (a) the anti-HER3 antibody-drug conjugate at a protein concentration of 1 to 40 mg / mL, (b) 0.05 to 0.6 mg / mL of polysorbate, (c) 10 to 150 mg / mL of sugar, and (d) 1 to 50 mM of acetate buffer, and the pH of the pharmaceutical composition is 4.9 to 5.5.

[0023] In an alternative embodiment, the pharmaceutical composition of any one of the above is The pharmaceutical composition contains (a) the anti-HER3 antibody-drug conjugate at a protein concentration of 1 to 40 mg / mL, (b) 0.05 to 0.6 mg / mL of polysorbate, (c) 10 to 150 mg / mL of sugar, and (d) 1 to 50 mM of acetate buffer, and the pH of the pharmaceutical composition is 5.0 to 5.5.

[0024] In an alternative embodiment, the pharmaceutical composition of any one of the above is The pharmaceutical composition contains (a) the anti-HER3 antibody-drug conjugate at a protein concentration of 1 to 40 mg / mL, (b) 0.05 to 0.6 mg / mL of polysorbate, (c) 40 to 120 mg / mL of sugar, and (d) 1 to 50 mM of acetate buffer, and the pH of the pharmaceutical composition is 5.0 to 5.5.

[0025] In an alternative embodiment, the pharmaceutical composition of any one of the above is The pharmaceutical composition contains (a) 10 to 30 mg / mL of the anti-HER3 antibody-drug conjugate, (b) 0.1 to 0.4 mg / mL of polysorbate, (c) 40 to 120 mg / mL of sugar, and (d) 5 to 15 mM of acetate buffer, and the pH of the pharmaceutical composition is 5.0 to 5.5.

[0026] In an alternative embodiment, the pharmaceutical composition of any one of the above is The pharmaceutical composition contains (a) the anti-HER3 antibody-drug conjugate at a protein concentration of 10 to 30 mg / mL, (b) 0.1 to 0.4 mg / mL of polysorbate, (c) 60 to 120 mg / mL of sugar, and (d) 5 to 15 mM of acetate buffer, and the pH of the pharmaceutical composition is 5.0 to 5.5.

[0027] In an alternative embodiment, the pharmaceutical composition of any one of the above is The pharmaceutical composition contains (a) the anti-HER3 antibody-drug conjugate at a protein concentration of 15 to 25 mg / mL, (b) 0.1 to 0.4 mg / mL of polysorbate 80, (c) 40 to 80 mg / mL of sucrose, and (d) 5 to 15 mM of acetic acid-sodium acetate buffer, and the pH of the pharmaceutical composition is 5.1 to 5.3.

[0028] In an alternative embodiment, the pharmaceutical composition of any one of the above is The pharmaceutical composition contains (a) the anti-HER3 antibody-drug conjugate at a protein concentration of 10 to 30 mg / mL, (b) 0.1 to 0.4 mg / mL of polysorbate, (c) 40 to 80 mg / mL of sugar, and (d) 5 to 15 mM of acetate buffer, and the pH of the pharmaceutical composition is 5.0 to 5.5.

[0029] In an alternative embodiment, the pharmaceutical composition of any one of the above is The pharmaceutical composition contains (a) the anti-HER3 antibody-drug conjugate at a protein concentration of 20 to 40 mg / mL, (b) 0.2 to 0.4 mg / mL of polysorbate 80, (c) 60 to 120 mg / mL of sucrose, and (d) 10 to 20 mM of acetic acid-sodium acetate buffer, and the pH of the pharmaceutical composition is 4.9 to 5.5.

[0030] In an alternative embodiment, the pharmaceutical composition of any one of the above is The pharmaceutical composition contains (a) the anti-HER3 antibody-drug conjugate at a protein concentration of about 20 mg / mL, (b) about 0.4 mg / mL of polysorbate 80, (c) about 120 mg / mL of sugar, and (d) about 10 mM of acetic acid-sodium acetate buffer, and the pH of the pharmaceutical composition is about 5.2.

[0031] In an alternative embodiment, the pharmaceutical composition of any one of the above is The pharmaceutical composition contains (a) the anti-HER3 antibody-drug conjugate at a protein concentration of about 20 mg / mL, (b) about 0.2 mg / mL of polysorbate 80, (c) about 60 mg / mL of sucrose, and (d) about 10 mM of acetic acid-sodium acetate buffer, and the pH of the pharmaceutical composition is about 5.2.

[0032] In an alternative embodiment, any one of the pharmaceutical compositions is a liquid formulation. The liquid or reconstituted formulations of the present disclosure have relatively good stability. Furthermore, stable liquid formulations include liquid formulations that exhibit desirable properties after storage for one month at temperatures including 40°C.

[0033] The present disclosure further provides a lyophilized formulation comprising an antibody-drug conjugate, which is capable of forming the above-described pharmaceutical composition after reconstitution, preferably with water for injection.

[0034] The present disclosure further provides a method for preparing a lyophilized formulation comprising an antibody-drug conjugate, the method comprising the step of lyophilizing the pharmaceutical composition described above.

[0035] In an alternative embodiment, a method for preparing a lyophilized formulation containing an antibody-drug conjugate includes the steps of pre-freezing, primary drying, and secondary drying. Lyophilization is achieved by sublimation of water at a temperature suitable for freezing the formulation and subsequent primary drying. Under these conditions, the product temperature is below the formulation's lower eutectic point or collapse temperature. Typically, the temperature range for primary drying is approximately -30 to 25°C (assuming the product remains frozen during the primary drying process). The drying time is determined by the formulation, the size and type of container (e.g., glass vial) containing the sample, and the volume of liquid, and may range from several hours to several days (e.g., 40 to 60 hours). The secondary drying step may be performed at approximately 0 to 40°C, and is primarily determined by the type and size of the container and the type of protein used. 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 moisture content of formulations lyophilized at low pressure is less than about 5%, preferably less than about 3%. The pressure may be the same as that used in the primary drying step, preferably the pressure for secondary drying is lower than that for primary drying. The conditions for lyophilization may vary depending on the formulation and vial size.

[0036] In one alternative embodiment of the present disclosure, the freeze-drying procedure is as follows.

[0037] (1) Pre-freeze at -2 to -8°C for 0.5 to 1 hour. (2) Pre-freeze at -45°C for 120 to 180 minutes. (3) Dry at -15 to -20°C and 0.1 mbar for 2100 to 2700 minutes. (4) Dry at 25°C and 0.05 mbar for 240 to 480 minutes.

[0038] In some embodiments, the lyophilized formulation is stable at 40° C. for at least 7 days, at least 14 days, at least 28 days, or at least 30 days.

[0039] The present disclosure further provides a lyophilized formulation comprising an antibody-drug conjugate, the lyophilized formulation being obtainable by lyophilizing the above-described pharmaceutical composition of the antibody-drug conjugate.

[0040] The present disclosure further provides a reconstituted solution containing an antibody-drug conjugate, the reconstituted solution being prepared by reconstituting the above-described lyophilized formulation.

[0041] The present disclosure further provides a method for preparing the reconstitution solution, the method comprising the step of reconstituting the lyophilized formulation, wherein the solution used for the reconstitution is selected from, but not limited to, water for injection, saline, or glucose solution.

[0042] In some embodiments, the pharmaceutical composition or reconstituted solution described above is an intravenous injection formulation, an intravenous injection formulation, a subcutaneous injection formulation, an intraperitoneal injection formulation, or an intramuscular injection formulation. In some embodiments, the pharmaceutical composition or reconstituted solution described above is an intravenous injection formulation.

[0043] In some embodiments, the pharmaceutical composition or reconstituted solution described above is applied for intravenous infusion, intravenous injection, subcutaneous injection, intraperitoneal injection or intramuscular injection, preferably for intravenous infusion.

[0044] In some embodiments, the pharmaceutical composition, reconstituted solution, or lyophilized formulation described above is used to prepare a drug for intravenous infusion, intravenous injection, subcutaneous injection, peritoneal injection, or intramuscular injection, and preferably, it is used to prepare a drug for intravenous infusion.

[0045] The present disclosure further provides an article of manufacture comprising a container containing the pharmaceutical composition, lyophilized formulation, or reconstitution solution described above. In some embodiments, the container is an injection bottle made of neutral borosilicate glass tubing.

[0046] The present disclosure further provides the use of the above pharmaceutical composition or lyophilized formulation or reconstituted solution or product in the preparation of a medicament for treating or preventing tumors.

[0047] The present disclosure further provides a method of treating a disease, comprising providing to a patient in need thereof the pharmaceutical composition or lyophilized formulation or reconstituted solution or product described above.

[0048] The present disclosure provides a pharmaceutical composition of an anti-HER3 antibody, wherein the anti-HER3 antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3 shown in SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, respectively, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3 shown in SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, respectively; The pharmaceutical composition further comprises an acetate buffer, preferably an acetic acid-sodium acetate buffer; The pharmaceutical composition has a pH of 5.0 to 5.5. The concentration of the antibody is 1 to 40 mg / mL, The sugar has a concentration of 10 to 100 mg / mL, and The concentration of the acetate buffer is 1 to 50 mM.

[0049] In an alternative embodiment, the pharmaceutical composition of any one of the above anti-HER3 antibodies, wherein the anti-HER3 antibody comprises a heavy chain variable region and a light chain variable region, The amino acid sequence of the heavy chain variable region is shown in SEQ ID NO:7, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO:8.

[0050] In an alternative embodiment, the anti-HER3 antibody pharmaceutical composition is any one of the above, wherein the anti-HER3 antibody comprises a heavy chain represented by SEQ ID NO: 9 and a light chain represented by SEQ ID NO: 10.

[0051] In an alternative embodiment, the pharmaceutical composition of any one of the above anti-HER3 antibodies further comprises a sugar. The "sugar" of the present disclosure is a sugar of the usual composition (CHO). n and derivatives thereof, including monosaccharides, disaccharides, trisaccharides, polysaccharides, sugar alcohols, reducing sugars, non-reducing sugars, etc. The sugar may be selected from glucose, sucrose, trehalose, α,α-trehalose dihydrate, lactose, fructose, maltose, dextran, glycerin, erythritol, glycerol, arabitol, sylitol, sorbitol, mannitol, melibiose, melezitose, raffinose, manninotriose, stachyose, maltose, lactulose, maltulose, sorbitol, maltitol, lactitol, iso-maltulose, etc. Preferred sugars are non-reducing disaccharides, more preferably α,α-trehalose dihydrate or sucrose, and most preferably sucrose.

[0052] In an alternative embodiment, in any one of the above pharmaceutical compositions of an anti-HER3 antibody, the sucrose concentration is 10 to 100 mg / mL, preferably 40 to 80 mg / mL, including, but not limited to, 10 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, 70 mg / mL, 80 mg / mL, 90 mg / mL, 100 mg / mL, and any range between these values, preferably 60 to 90 mg / mL, more preferably about 60 mg / mL. The term "about" above is most preferably ±10%, and for example, the sucrose concentration is about 60 mg / mL, i.e., 60 mg / mL ±6 mg / mL.

[0053] In an alternative embodiment, in any one of the above pharmaceutical compositions of an anti-HER3 antibody, the buffer concentration is about 1 to about 50 mM, including, but not limited to, 1 mM, 5 mM, 8 mM, 9 mM, 10 mM, 11 mM, 12 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 30 mM, 40 mM, 50 mM, and any range between these values, preferably 5 to 15 mM, more preferably about 10 mM. The term "about" above most preferably refers to ±10%, and for example, the concentration of the acetic acid-sodium acetate buffer is about 10 mM, i.e., 10 mM ± 1 mM.

[0054] In an alternative embodiment, in any one of the above pharmaceutical compositions of an anti-HER3 antibody, the concentration of the anti-HER3 antibody is 1 to 40 mg / mL, and non-limiting examples include 1 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, 40 mg / mL, and any range between these point values. Preferably, the concentration of the anti-HER3 antibody is 10 to 30 mg / mL, more preferably, the concentration of the anti-HER3 antibody is 18 to 22 mg / mL, and most preferably, about 20 mg / mL. Specific, non-limiting examples include 20.1 mg / mL, 20.2 mg / mL, 20.3 mg / mL, 20.4 mg / mL, 20.5 mg / mL, 20.6 mg / mL, 20.7 mg / mL, 20.8 mg / mL, 20.81 mg / mL, 20.82 mg / mL, 20.83 mg / mL, 20.84 mg / mL, 20.85 mg / mL, 20.86 mg / mL, 20.87 mg / mL, 20.91 mg / mL, 20.92 mg / mL, 20.93 mg / mL, 20.94 mg / mL, 20.95 mg / mL, 20.96 mg / mL, 20.97 mg / mL, 20.98 mg / mL, 20.99 mg / mL, 21 mg / mL, and any range between these point values. The term "about" above is most preferably 10%, e.g., a protein concentration of about 20 mg / mL, i.e., 20±10 mg / mL, is most preferably.

[0055] In an alternative embodiment, a pharmaceutical composition of any one of the above anti-HER3 antibodies is provided, which comprises: (a) an anti-HER3 antibody having a concentration of 10 to 30 mg / mL, (b) a sugar having a concentration of 60 to 90 mg / mL, and (c) an acetate buffer having a concentration of 5 to 15 mM, wherein the pharmaceutical composition has a pH of 5.1 to 5.4; In an alternative embodiment, a pharmaceutical composition of any one of the above anti-HER3 antibodies is provided, which comprises: (a) an anti-HER3 antibody having a concentration of 10 to 30 mg / mL, (b) a sugar having a concentration of 40 to 80 mg / mL, and (c) an acetate buffer having a concentration of 5 to 15 mM, wherein the pH of the pharmaceutical composition is 5.1 to 5.4; In an alternative embodiment, a pharmaceutical composition of any one of the above anti-HER3 antibodies is provided, which comprises: The pharmaceutical composition comprises (a) the anti-HER3 antibody at a concentration of about 20 mg / mL, (b) about 60 mg / mL sucrose, and (c) about 10 mM acetic acid-sodium acetate buffer, and the pH of the pharmaceutical composition is about 5.2.

[0056] The present disclosure further provides a pharmaceutical composition of the anti-HER3 antibody-drug conjugate, or a pharmaceutical composition of the anti-HER3 antibody, or a lyophilized preparation, or a reconstituted solution, or a product as a medicament for use in treating a disease.

[0057] The present disclosure further provides a pharmaceutical composition of the anti-HER3 antibody-drug conjugate, or a pharmaceutical composition of the anti-HER3 antibody, or a lyophilized preparation, or a reconstituted solution, or a product as a medicament for use in treating tumors or cancer.

[0058] In an alternative embodiment, the disease is a disease or condition mediated by HER3.

[0059] The present disclosure further provides use of the above-mentioned anti-HER3 antibody-drug conjugate pharmaceutical composition, or anti-HER3 antibody pharmaceutical composition, or lyophilized formulation, or reconstituted solution, or product as a drug in the preparation of a medicament for treating viral infection, tumor, or cancer, wherein the above-mentioned tumor or cancer is selected from breast cancer, non-small cell lung cancer, gastric cancer, ovarian cancer, prostate cancer, bladder cancer, colorectal cancer, head and neck squamous cell carcinoma, and melanoma.

[0060] When point values ​​are referred to in this disclosure, they should be understood to include a margin of error due to factors such as laboratory environment, operator manipulation, equipment, methodology, and measurement error.

[0061] As will be appreciated by those skilled in the art, one, some, or all of the features of each embodiment described in the present disclosure can be further combined to form other embodiments of the present disclosure. These and other combined embodiments of the present disclosure are further illustrated in the detailed description below. [Means for solving the problem]

[0062] The present disclosure provides pharmaceutical compositions that are more convenient to manufacture and administer and have stable performance. Among these, undesirable instability may include any one or more of 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 cysteine, toxin release, etc. Specifically, the pharmaceutical compositions described in the present disclosure comprise an antibody-drug conjugate and a buffer.

[0063] term In order that the present disclosure may be more readily understood, certain technical and scientific terms are specifically defined below. Unless expressly defined otherwise herein, all other technical and scientific terms used herein have the meanings commonly understood by those of ordinary skill in the art.

[0064] An "antibody drug conjugate (ADC)" is an antibody linked via a linker unit to a biologically active cytotoxin or cell-killing small molecule drug.

[0065] The "drug loading" or "drug load," also referred to as the drug-to-antibody ratio (DAR), refers to the average number of drugs coupled to each antibody in an ADC. For example, it may be within a range of about 1 to about 10 drugs conjugated to each antibody. In some embodiments, it may be within a range of about 1 to about 8 drugs conjugated to each antibody, preferably 2 to 8, 2 to 7, 2 to 6, 2 to 5, 2 to 4, 3 to 4, 3 to 5, 5 to 6, 5 to 7, 5 to 8, or 6 to 8. For example, the drug loading may be an average of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. The general formula of an ADC according to the present disclosure includes a set of antibodies coupled to drugs within the above range. In embodiments of the present disclosure, the drug loading may be represented by n. Drug loading can be measured by conventional methods, such as UV / visible spectroscopy, mass spectrometry, ELISA, and HPLC.

[0066] The term "linker unit" or "linking fragment" or "linking unit" refers to a chemical structural fragment or bond that is linked at one end to an antibody or antigen-binding fragment thereof and at the other end to a drug, and may be linked to another linker before being linked to a drug.

[0067] The linker may comprise an extender, a spacer, and an amino acid unit and may be synthesized by methods known in the art, such as those described in US20050238649A1. The linker may be a "cleavable linker" that facilitates drug release in cells. For example, an acid-labile linker (e.g., hydrazone), a protease-sensitive (e.g., 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. US5208020A) may be used.

[0068] The term "drug-linked arm fragment" or "drug-linker fragment" refers to a fragment in which a drug is linked to a linker unit, and can be linked to an antibody via the other end of the linker unit.

[0069] The loading of the cytotoxic drug is (1) controlling the molar ratio of the drug-linking arm fragment to the monoclonal antibody; (2) controlling the reaction time and temperature; (3) selecting different reaction reagents; The amount of oxygen can be controlled by a number of methods, including but not limited to:

[0070] The three-letter and one-letter codes for amino acids used in this disclosure are as described in J. Biol. Chem, 243, p. 3558 (1968).

[0071] The term "antibody" as used in this disclosure is used in the broadest sense and covers a variety of antibody structures, including, but not limited to, full-length antibodies and antibody fragments (or antigen-binding fragments or antigen-binding portions), provided they exhibit the desired antigen-binding activity. Typically, naturally occurring intact antibodies are tetrapeptide chain structures consisting of two identical heavy chains and two identical light chains linked by interchain disulfide bonds.

[0072] The engineered antibodies or antigen-binding fragments of the present disclosure can be prepared and purified by conventional methods. For example, 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. In one preferred conventional technique, a mammalian expression system induces antibody glycosylation, particularly at the highly conserved N-terminal site of the Fc region. Positive clones are expanded in serum-free medium in a bioreactor to produce antibodies. The culture medium from which the antibodies are secreted can be purified by conventional techniques, for example, on an A or G Sepharose FF column containing a conditioned buffer. Nonspecifically bound components are washed away. The bound antibodies can then be eluted using a pH gradient, and the antibody fragments detected and collected by SDS-PAGE. The antibodies can be concentrated by filtration using conventional methods. Soluble mixtures and multimers can be removed by conventional methods, such as molecular sieving or ion exchange. The resulting product must be immediately frozen, such as at -70°C, or lyophilized.

[0073] "Buffer" refers to a buffer that resists changes in pH by the action of its acid-base conjugate components. Examples of buffers that control pH within an appropriate range include acetate, succinate, gluconate, histidine, oxalate, lactate, phosphate, citrate, citric acid-sodium citrate, tartrate, fumarate, glycylglycine, and other organic acid buffers.

[0074] A "histidine buffer" is a buffer containing histidine. Examples of histidine buffers include histidine-histidine hydrochloride, histidine-histidine acetate, histidine-histidine phosphate, and histidine-histidine sulfate, with histidine-histidine hydrochloride being preferred. Histidine-histidine hydrochloride buffers may be prepared from histidine and hydrochloric acid, or from histidine and histidine hydrochloride.

[0075] A "citrate buffer" is a buffer containing citrate ions. Examples of citrate buffers include citric acid-sodium citrate, citric acid-potassium citrate, citric acid-calcium citrate, citric acid-magnesium citrate, etc. A preferred citrate buffer is citric acid-sodium citrate.

[0076] A "succinate buffer" is a buffer containing succinate ions. Examples of succinate buffers include succinic acid-sodium salt, succinic acid-potassium succinate, succinic acid-calcium salt, etc. A preferred succinate buffer is succinic acid-sodium salt. Illustratively, the succinic acid-sodium succinate may be prepared from succinic acid and sodium hydroxide, or from succinic acid and sodium succinate salt.

[0077] A "phosphate buffer" is a buffer containing phosphate ions. Examples of phosphate buffers include disodium hydrogen phosphate-sodium dihydrogen phosphate, disodium hydrogen phosphate-potassium dihydrogen phosphate, disodium hydrogen phosphate-citric acid, etc. A preferred phosphate buffer is disodium hydrogen phosphate-sodium dihydrogen phosphate.

[0078] An "acetate buffer" is a buffer containing acetate ions. Examples of acetate buffers include acetic acid-sodium acetate, histidine-histidine acetate, acetic acid-potassium acetate, acetic acid-calcium acetate, acetic acid-magnesium acetate, etc. A preferred acetate buffer is acetic acid-sodium acetate.

[0079] A "pharmaceutical composition" refers to a mixture containing one or more antibody-drug conjugates described herein or physiologically / pharmaceutical acceptable salts or prodrugs thereof, together with other chemical components, such as physiologically / pharmaceutically acceptable carriers and excipients. The pharmaceutical composition is intended to maintain the stability of the antibody active ingredient, facilitate administration to the body, and contribute to the absorption of the active ingredient to further exert its biological activity.

[0080] In this disclosure, "pharmaceutical composition" and "formulation" are not mutually exclusive.

[0081] Unless otherwise specified, the solvent in any solution form of the pharmaceutical composition described in the present disclosure is water.

[0082] "Lyophilized formulation" refers to a pharmaceutical composition in liquid or solution form or a formulation or pharmaceutical composition obtained after a freeze-drying step is performed on a liquid or solution formulation.

[0083] Those skilled in the art should understand that when used with reference to a numerical range, cutoff value, or specific value, "about" can refer to within one or more standard deviations. Alternatively, "about" can refer to a range with up to a 20% difference (i.e., ±20%). Because many of the numerical values ​​used herein were determined experimentally, those skilled in the art should understand that such determinations can and usually do vary between different experiments. Due to this inherent variation, it is believed that the values ​​used herein should not be unduly limited. Thus, the term "about" is used to cover a variation of ±20% or less, a variation of ±10% or less, a variation of ±5% or less, a variation of ±1% or less, a variation of ±0.5% or less, or a variation of ±0.1% or less from the specified value.

[0084] The pharmaceutical compositions described herein can achieve stable efficacy, i.e., the antibody-drug conjugate therein essentially retains its physical stability and / or chemical stability and / or biological activity after storage. Preferably, the pharmaceutical composition essentially retains its physical and chemical stability and biological activity after storage. The storage period is generally selected based on the intended shelf life of the pharmaceutical composition. Currently, there are several analytical techniques for measuring the stability of proteins or antibody-drug conjugates, which can be used to measure stability after storage at a predetermined temperature for a predetermined period of time.

[0085] A stable formulation is one that shows no significant change when stored at refrigerated temperatures (2-8°C) for at least 6 months, preferably 12 months, and more preferably 2 years. Stable liquid formulations also include liquid formulations that exhibit desired characteristics after storage at temperatures including 25°C for periods including 1 month, 3 months, and 6 months. Furthermore, stable liquid formulations include liquid formulations that exhibit desired characteristics after storage at temperatures including 40°C for periods including 10 days, 20 days, and 1 month. Typical examples of stability include those measured by SEC-HPLC in which aggregation or degradation of antibody monomers typically occurs at no more than about 10%, preferably no more than about 5%. Visual analysis reveals that the formulation is a pale yellow, almost colorless, transparent liquid, or colorless, or clear to slightly opalescent. The concentration, pH, and osmolality of the formulation vary by no more than ±10%. A decrease of no more than about 10%, preferably no more than about 5%, is typically observed. No more than about 10%, preferably no more than about 5%, of the antibody monomers typically forms aggregates.

[0086] An antibody-drug conjugate "retains its physical stability" in a drug formulation if it does not exhibit significant increased aggregation, precipitation, and / or denaturation, as determined by visual color and / or clarity or 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 protein tertiary structure) and FTIR spectroscopy (which determines protein secondary structure).

[0087] An antibody-drug conjugate "retains its chemical stability" in a drug formulation if it does not undergo significant chemical changes. Chemical stability can be assessed by detecting and quantifying chemically altered forms of the protein. Degradation processes that frequently alter the chemical structure of proteins include hydrolysis or cleavage (e.g., assessed by methods such as size exclusion chromatography and CE-SDS), oxidation (e.g., assessed by methods such as peptide mapping coupled with mass spectrometry or MALDI / TOF / MS), deamidation (e.g., assessed by methods such as ion exchange chromatography, capillary isoelectric focusing, peptide mapping, or isoaspartic acid content measurement), and isomerization (e.g., assessed by isoaspartic acid content measurement, peptide mapping, etc.).

[0088] An antibody-drug conjugate "retains its biological activity" in a drug formulation if the biological activity at a given time is within a predetermined range of the biological activity exhibited when the drug formulation is prepared.

[0089] "Optional" or "optionally" means that the subsequently described event or circumstance may occur, but does not necessarily occur, and the description includes both cases where the event or circumstance occurs and cases where it does not. For example, "optionally comprising 1 to 3 antibody heavy chain variable regions" means that antibody heavy chain variable regions of a particular sequence may be present, but are not necessarily present.

[0090] The term "substituted" refers to one or more hydrogen atoms in a group, preferably up to 5, more preferably 1 to 3 hydrogen atoms, being independently replaced with a corresponding number of substituents. Of course, substituents are only located at their chemically feasible positions, and those skilled in the art can determine (by experiment or theory) possible or impossible substitutions without much effort. For example, an amino group or a hydroxy group having free hydrogen may be unstable if it is bonded to a carbon atom having an unsaturated (e.g., olefinic) bond.

[0091] The preparation of conventional pharmaceutical compositions is set out in the Chinese Pharmacopoeia.

[0092] The term "carrier," as used in the present disclosure, refers to a system that can change the drug's entry into and distribution within the human body, control the drug's release rate, and transport the drug to target organs. The drug carrier's release and targeting system can reduce drug degradation and loss, lower side effects, and improve bioavailability. For example, polymeric surfactants, which are used as carriers, can self-assemble to form various types of aggregates due to their unique amphiphilic structure, and preferred examples include micelles, microemulsions, gels, liquid crystals, and vesicles. These aggregates have the ability to encapsulate drug molecules and good membrane permeability, making them good drug carriers.

[0093] "Administration" and "treatment," when applied to an animal, human, or experimental subject, cell, tissue, organ, or biological fluid, refer to the contact of an exogenous agent, therapeutic agent, diagnostic agent, or composition with an animal, human, subject, cell, tissue, organ, or biological fluid. "Administration" and "treatment" can refer, for example, to therapeutic, pharmacokinetic, diagnostic, research, and experimental methods. Treatment of a cell includes contact of a reagent with the cell and contact of a reagent with a fluid, where the fluid contacts the cell. "Administration" and "treatment" also refer to ex vivo and in vitro treatment, e.g., of a cell, with a reagent, diagnostic, binding composition, or via another cell. "Treatment," when applied to a human, veterinary, or research subject, refers to therapeutic treatment, preventative or prophylactic measures, and research and diagnostic uses.

[0094] "Treatment" refers to the administration of an oral or topical therapeutic agent, such as a composition comprising any one of the binding compounds of the present disclosure, to a patient having one or more disease symptoms for which the known therapeutic agent has a therapeutic effect. Typically, the patient or population being treated is administered an amount of therapeutic agent that effectively alleviates one or more disease symptoms, thereby inducing regression of such symptoms or inhibiting the progression of such symptoms to any clinically measurable extent. The amount of therapeutic agent that effectively alleviates any particular disease symptom (also referred to as a "therapeutically effective amount") can vary depending on various factors, including the patient's disease state, age, and weight, and the ability of the drug to produce the desired therapeutic effect in the patient. Reduction of disease symptoms can be assessed by any clinical detection method commonly used by physicians or other professional health care providers to assess the severity or progression of the condition. Although an embodiment of the present disclosure (e.g., a method of treatment or product) may be ineffective in alleviating each target disease symptom, it should reduce the target disease symptom in a statistically significant number of patients, as determined by any statistical testing method known in the art, such as Student's t-test, chi-square test, Mann and Whitney U test, Kruskal-Wallis test (H test), Jonckheere-Terpstra test, and Wilcoxon test.

[0095] An "effective amount" includes an amount sufficient to ameliorate or prevent the symptoms or conditions of a medical disorder. An effective amount also refers to an amount sufficient to enable or facilitate diagnosis. The effective amount used in a particular patient or veterinary subject can vary depending on factors such as the condition being treated, the patient's overall health, the route and dose of administration, and the severity of side effects. An effective amount may be the maximum dose or dosing regimen that avoids significant side effects or toxic effects.

[0096] "Replacement" refers to the replacement of a solvent system that dissolves the antibody protein or antibody-drug conjugate, for example, replacing a high-salt or high-osmolarity solvent system containing the antibody protein or antibody-drug conjugate with a buffer system of a stable formulation by a physical manipulation method such that the antibody protein or antibody-drug conjugate is present in a stable formulation. Such physical manipulation methods include, but are not limited to, ultrafiltration, dialysis, or centrifugation. DETAILED DESCRIPTION OF THE INVENTION

[0097] The present disclosure will be further described below in conjunction with examples, but these examples are not intended to limit the scope of the present disclosure. In the examples of the present disclosure, experimental methods for which specific conditions are not specified generally follow standard conditions, such as those in the "Antibody Technology Laboratory Manual" and "Molecular Cloning Manual" published by Cold Spring Harbor Laboratory, or the conditions recommended by raw material or product manufacturers. Reagents for which specific sources are not specified are standard commercially available reagents. [Example]

[0098] Preparation Example 1: Preparation of antibody The HER3 antibody of the present application is derived from antibody HER3-29 of WO2022078425A1 (which is incorporated herein by reference in its entirety), and has the following relevant sequences:

[0099] 1) The sequences of the anti-HER3 antibody CDRs are shown in the table below.

[0100] [Table 1] 2) Fully human antibody molecule The heavy chain variable region and light chain variable region sequences of the anti-HER3 antibody are as follows:

[0101] HER3 heavy chain variable region:

number

number

[0102] HER3-29 heavy chain:

number

number

[0103] Preparation Example 2, Preparation of ADC The drug moiety of the conjugates of the present disclosure may be any suitable drug. Particularly suitable drugs are described, for example, in PCT Publication No. WO2020063676A1, which is incorporated herein by reference in its entirety.

[0104] Compound 2-B of the present disclosure (i.e., compound 2-B of Example 2 of WO2020063676 A1) is (R)-2-cyclopropyl-N-((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-1-yl)-2-hydroxyacetamide, and has the following structure:

[0105] [ka] Compound 9A of the present disclosure (i.e., compound 9-A of Example 9 of WO2020063676 A1) is N-((2R,10S)-10-benzyl-2-cyclopropyl-1-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl)amino)-1,6,9,12,15-pentaoxo-3-oxa-5,8,11,14-tetraazahexadecan-16-yl)-6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamide, having the following structure:

[0106] [ka] Exemplary anti-HER3 antibody-drug conjugates of the present disclosure are Examples 3-1 (n=4.19), 3-2 (n=2.91), and 3-3 (n=7.27) in WO2022078425A1, and have the following structures:

[0107] [ka] Among them, n is 1 to 8, preferably 3 to 5.

[0108] The structure of the pharmaceutical conjugate molecule in this disclosure is abbreviated as structural formula ADC-1 below.

[0109] [ka] The best example of the formulation in this disclosure has an n (DAR value) of about 4.0, which corresponds to 4.0±0.4.

[0110] Preparation Example 3: Preparation of formulation The equipment, parameters and calculation methods used in the preparation and detection process of the formulation are as follows:

[0111] 1)SEC-HPLC purity: Monomers and polymers are isolated based on the molecular size of the components to be measured by size exclusion chromatography (SEC) according to the molecular sieving mechanism of a gel chromatography column. The ratio of monomers to polymers is calculated by the area normalization method at a detection wavelength of 280 nm.

[0112] Equipment for SEC measurements: Agilent 1260 high performance liquid chromatograph.

[0113] Column: Tosoh, TSKgel G3000SWXL (7.8 mm × 30 cm, 5 μm).

[0114] 2) CE-SDS purity (reduced CE-SDS and non-reduced CE-SDS purity): The purity of reduced and non-reduced CE-SDS is measured by capillary gel electrophoresis. Using a capillary as the isolation channel and a high-voltage DC electric field as the driving force, separation is achieved based on the differences in the mobility (migration speed at a unit electric field strength) and / or partitioning behavior of each component in the sample.

[0115] Equipment for CE-SDS measurement: Capillary electrophoresis apparatus, model number Beckman / PA800 plus.

[0116] ΔR-CE% represents the difference between the detection item after placing the sample under each condition and when the sample was first placed.

[0117] 3) Measurement of protein concentration: The concentration of the antibody-drug conjugate in this disclosure is calculated as the protein concentration, i.e., is expressed as the concentration of the antibody moiety in the antibody-drug conjugate.

[0118] The toxin in the antibody-drug conjugate is absorbed at the protein's characteristic absorption wavelength of 280 nm, and also absorbs at 370 nm. Therefore, the concentration of the protein is calculated using the following formula:

number

[0119] When the test solution is diluted, the protein concentration JPEG2025539862000012.jpg1399, where N is the dilution factor.

[0120] Equipment for measuring protein concentration: UV-visible spectrophotometer, model number: Nano Drop 2000.

[0121] 4) Appearance: By visual inspection, wipe the sample bottle clean and directly observe the appearance of the freeze-dried solid powder. Also, observe the color, transparency and visible foreign matter of the sample solution under the white background and black background of the transparency detector, respectively, at a light irradiation intensity of 1500 lx.

[0122] Appearance detection equipment: Seitakuki YB-2A transparency detector.

[0123] 5) Thermal decomposition onset temperature (Tonset) and particle size (Radius): The thermal decomposition onset temperature (Tonset) and particle size (Radius) of proteins are measured using a high-throughput dynamic light scattering meter. The sample is placed on a sample plate, the particle size and dispersion coefficient (%PD) are measured, and Tonset is measured by heating the sample from 25 to 95°C.

[0124] Tonset and Radius measurement equipment: High-throughput dynamic light scattering meter, model number DynaPro plate Reader III.

[0125] 6) Charge variant content: The content of charge variants was measured using the iCIEF analytical detection method. A capillary was used as the isolation channel, and a DC voltage was applied to both ends of the capillary. The ampholyte solution in the capillary formed a pH gradient over a certain range, and each component migrated to its respective isoelectric point based on the difference in charge and was then focused into a very narrow compartment, thereby achieving isolation of the components.

[0126] iCIEF measurement equipment: Protein Simple full-column imaging capillary isoelectric focusing system, model number iCE3 or Maurice.

[0127] 7) DAR value: The drug-antibody ratio (DAR) is measured by hydrophobic chromatography (HIC method). The hydrophobicity of components with different DAR values ​​is measured using a hydrophobic chromatography column. The sample is isolated based on the hydrophobicity of the components to be measured. The peak area of ​​each peak is calibrated at a detection wavelength of 280 nm, and then substituted into the calculation formula to obtain the DAR value.

[0128] Instrument for HIC method measurement: Agilent 1260 high performance liquid chromatograph.

[0129] Column: BioCore HIC-Butyl.

[0130] 8) Free toxins: After protein precipitation, the supernatant is taken and measured by UPLC-UV method.

[0131] Measurement equipment: Waters ultra-high performance liquid chromatograph, model number ACQUITY H-Class.

[0132] Exemplary antibody pharmaceutical composition (formulation) preparation process Step 1: Prepare a bulk formulation containing the HER3 antibody-drug conjugate ADC-1 (DAR value 4.0±0.4) and a stabilizer, sterilize it through a 0.22 μm filter, and collect the filtrate.

[0133] Step 2: Adjust the filling volume (target filling volume is 5.3 mL / bottle), fill the collected filtrate into vials, and sample at the start, middle, and end of filling to detect any differences in the filling volume.

[0134] Step 3: Put half of the plugs in, freeze-dry, put all the plugs in, open the capping device and cap.

[0135] Step 4: Visually inspect the product to ensure there are no cosmetic defects such as crumbling, shrinkage, or melting of the solid powder. Print the carton label, fold the carton, pack the product, and label the carton.

[0136] The lyophilization process for the ADC formulation is as follows:

[0137] (1) Pre-freeze at -2 to -8°C for 0.5 to 1 hour. (2) Pre-freeze at -45°C for 120 to 180 minutes. (3) Dry at -15 to -20°C and 0.1 mbar for 2100 to 2700 minutes. (4) Dry at 25°C and 0.05 mbar for 240 to 480 minutes.

[0138] Example 1. pH screening of antibody HER3-29 A 10 mM acetic acid-sodium acetate buffer system was selected, and six different pH values ​​(3.7, 4.6, 5.0, 5.4, 5.8, and 6.2) were designed. Six formulations with a HER3-29 antibody concentration of 25.0 mg / mL were prepared. The thermal decomposition onset temperature (Tonset) and particle size (Radius) of the samples were measured, and the stability of the protein under different pH conditions was investigated.

[0139] 1) 10 mM acetic acid-sodium acetate, pH 3.7; 2) 10 mM acetic acid-sodium acetate, pH 4.6; 3) 10 mM acetic acid-sodium acetate, pH 5.0; 4) 10 mM acetic acid-sodium acetate, pH 5.4; 5) 10 mM acetic acid-sodium acetate, pH 5.8, and 6) 10 mM acetic acid-sodium acetate, pH 6.2.

[0140] [Table 2]

[0141] The results showed that antibody HER3-29 had a relatively small particle size and a relatively high Tonset value at pH 5.0, and the protein was less likely to aggregate at this pH condition. Considering that the pH range is usually within ±0.2 of the target value and that the Tonset value is higher at pH >5.0 than at pH <5.0, a pH of 5.2 ±0.2 was preferred for antibody HER3-29.

[0142] Example 2. Confirmation of pH of antibody HER3-29 A 10 mM acetic acid-sodium acetate buffer solution at pH 5.2 was used to prepare a formulation containing 20 mg / mL of HER3-29 antibody. The stability of the formulation was examined at high temperatures (40°C), 25°C, 2-8°C, -35°C, and repeated freeze-thaw cycles (-35°C and room temperature) by detecting the sample's appearance, SEC-HPLC, and iCIEF purity.

[0143] 10 mM acetic acid-sodium acetate, pH 5.2, 20 mg / mL antibody HER3-29.

[0144] [Table 3] [Table 4]

[0145] Note: TO represents day 0, 1M represents 1 month, 1W represents 1 week, and so on.

[0146] In a 10 mM acetic acid-sodium acetate buffer system, all formulation samples remained colorless under different storage conditions, but freeze-thawing slightly increased opalescence. Storage at 2-8°C and 25°C showed no significant changes in the polymer and monomer content of the samples. However, storage at -35°C and freeze-thawing tended to increase the polymer content, and at a higher temperature of 40°C, the polymer decomposed into some monomers and fragments. Storage at -35°C and 2-8°C increased the acidic peak, while the main and basic peaks decreased, but the changes were not significant. Freezing and thawing did not significantly affect the charge heterogeneity of the samples. The HER3-29 antibody showed a significant increase in polymer content after storage at -35°C and freeze-thawing, but remained relatively stable at 2-8°C and 25°C. Considering that the addition of stabilizers may affect the subsequent coupling reaction, the target pH for antibody HER3-29 was selected to be 5.2 without the addition of stabilizers.

[0147] Example 3. Screening of buffer systems for antibody HER3-29 Four buffer systems were selected: 10 mM histidine-histidine hydrochloride at pH 5.2, 10 mM citric acid-sodium citrate, 10 mM succinic acid-sodium succinate, and 10 mM acetic acid-sodium acetate. The stability of the antibody HER3-29 at 20 mg / mL was investigated by detecting the sample appearance, SEC-HPLC, and iCIEF purity at high temperatures (40°C), 25°C, 2-8°C, and -35°C, repeated freeze-thaw cycles (-35°C and room temperature), shaking (200 rpm, 25°C), and light exposure (25°C, 5000 lx).

[0148] 1) 10 mM histidine-histidine hydrochloride, pH 5.2, 20 mg / mL antibody HER3-29; 2) 10 mM citric acid-sodium citrate, pH 5.2, 20 mg / mL antibody HER3-29; 3) 10 mM succinic acid-sodium succinate, pH 5.2, 20 mg / mL antibody HER3-29, and 4) 10 mM acetic acid-sodium acetate, pH 5.2, 20 mg / mL antibody HER3-29.

[0149] [Table 5] [Table 6-1] [Table 6-2] [Table 7]

[0150] Note: NT indicates not detected. For formulations 1 to 3, the content of the basic peak was relatively high and the content of the main peak was relatively low under different conditions, so some samples were not detected under certain conditions.

[0151] The results showed that the antibody HER3-29 was colorless in all different buffer systems and showed slight opalescence in the 10 mM citric acid-sodium citrate system. Compared to 0°C, the polymer content of each formulation did not significantly increase under conditions of 2-8°C, shaking, and light exposure. However, the polymer content increased under conditions of -35°C and repeated freeze-thawing. At a high temperature of 40°C, the polymer decomposed into some monomers and fragments, resulting in a decrease in polymer content. In the 10 mM histidine-histidine hydrochloride, 10 mM citric acid-sodium citrate, and 10 mM succinic acid-sodium succinate buffer systems, the content of the basic peak was significantly higher than in the 10 mM acetic acid-sodium acetate buffer system. In the 10 mM acetic acid-sodium acetate buffer system, the content of the main peak decreased to some extent under high temperature conditions, but no significant changes were observed under other conditions. Therefore, the buffer system for antibody HER3-29 was selected to be 10 mM acetic acid-sodium acetate at pH 5.2.

[0152] Example 4. Screening for stabilizers of antibody HER3-29 According to the results of the previous screening, the polymer content of the antibody HER3-29 increased in a 10 mM acetic acid-sodium acetate buffer system at pH 5.2 under conditions of -35°C and repeated freezing and thawing. Prior to coupling, the antibody HER3-29 needed to be frozen at -35°C. To enhance the frozen storage and freeze-thaw stability of the antibody HER3-29, a 10 mM acetic acid-sodium acetate buffer system at pH 5.2 was selected. Two formulation samples were prepared, each containing 60 mg / mL sucrose as a stabilizer, with or without 0.2 mg / mL polysorbate 80, and each containing 20 mg / mL of antibody HER3-29. By detecting the appearance, SEC-HPLC, and iCIEF purity of the samples, the stability of the two formulations was investigated under high temperature (40°C), 25°C, 2-8°C, -35°C, repeated freeze-thawing (-35°C and room temperature), shaking (200 rpm, 25°C), and light exposure (25°C, 5000 lx).

[0153] 1) 10 mM acetic acid-sodium acetate, 60 mg / mL sucrose, pH 5.2, 20 mg / mL antibody HER3-29, and 2) 10 mM acetic acid-sodium acetate, 60 mg / mL sucrose, 0.2 mg / mL polysorbate 80, pH 5.2, 20 mg / mL antibody HER3-29.

[0154] [Table 8] [Table 9] [Table 10]

[0155] As a result, at a high temperature of 40°C, the main peak content of both formulations decreased to some extent, and the polymer content decreased slightly due to the decomposition of the polymer into some monomers and fragments. Both formulations showed good stability at 25°C, 2-8°C, -35°C, repeated freeze-thawing (-35°C and room temperature), shaking (200 rpm, 25°C), and light exposure (25°C, 5000 lx). There was no significant difference in stability between the two formulations under each of the conditions examined. This indicates that the addition of 60 mg / mL sucrose to a 10 mM acetic acid-sodium acetate buffer system at pH 5.2, with or without the addition of 0.2 mg / mL polysorbate 80, can improve the cryopreservation and freeze-thaw stability of the antibody HER3-29.

[0156] Example 5. Screening of stabilizer concentrations for antibody HER3-29 A 10 mM acetic acid-sodium acetate buffer system at pH 5.0 or pH 5.4 was selected, and 60 mg / mL or 90 mg / mL sucrose was used as a stabilizer. A total of four formulations were prepared, each with an antibody HER3-29 concentration of 20 mg / mL. The stability of the four formulations was examined under high temperature (40°C), repeated freeze-thawing (-35°C and room temperature), shaking (200 rpm, 25°C), and light exposure (25°C, 5000 lx) conditions by detecting the sample appearance and SEC-HPLC purity.

[0157] 1) 10 mM acetic acid-sodium acetate, 60 mg / mL sucrose, pH 5.0, 20 mg / mL antibody HER3-29; 2) 10 mM acetic acid-sodium acetate, 90 mg / mL sucrose, pH 5.0, 20 mg / mL antibody HER3-29; 3) 10 mM acetic acid-sodium acetate, 60 mg / mL sucrose, pH 5.4, 20 mg / mL antibody HER3-29, and 4) 10 mM acetic acid-sodium acetate, 90 mg / mL sucrose, pH 5.4, 20 mg / mL antibody HER3-29.

[0158] [Table 11] [Table 12]

[0159] As a result, the appearance of the four formulations did not change significantly under each of the different test conditions. At a high temperature of 40°C, the polymer content decreased slightly due to decomposition of the polymer into some monomers and fragments. However, the monomer and polymer contents did not change significantly under the other test conditions and time points, demonstrating that antibody HER3-29 exhibited good stability in these four systems. Considering that polysorbate 80 may affect the subsequent coupling process, the system for antibody HER3-29 was selected to be 10 mM acetic acid / sodium acetate, 60-90 mg / mL sucrose, pH 5.0-5.4 (pH 5.2±0.2).

[0160] Example 6. Protein concentration screening of antibody HER3-29 A 10 mM acetic acid-sodium acetate buffer system at pH 5.2 was selected, and 60 mg / mL sucrose was used as a stabilizer. Two sets of formulations were prepared, each with an antibody HER3-29 concentration of 20 mg / mL or 40 mg / mL. The stability of the two sets of formulations was examined under high temperature (40°C), 25°C, 2-8°C, -35°C, repeated freeze-thawing (-35°C and room temperature), shaking (200 rpm, 25°C), and light exposure (25°C, 5000 lx) conditions by detecting the sample appearance, SEC-HPLC, iCIEF, and NR-CE purity.

[0161] 1) 10 mM acetic acid-sodium acetate, 60 mg / mL sucrose, pH 5.2, 20 mg / mL antibody HER3-29, and 2) 10 mM acetic acid-sodium acetate, 60 mg / mL sucrose, pH 5.2, 40 mg / mL antibody HER3-29.

[0162] [Table 13] [Table 14] [Table 15] [Table 16-1] [Table 16-2]

[0163] As a result, the two formulations showed no significant differences in SEC, iCIEF, and NR-CE purity under different conditions. However, when the antibody HER3-29 concentration was 40 mg / mL, small amounts of protein particles appeared in all samples under high temperature conditions of 40°C, 25°C, and 10 days (10d) of light irradiation. Therefore, the concentration of antibody HER3-29 was selected to be 20 mg / mL.

[0164] Example 7. pH Screening of ADC-1 Three formulations containing 20.0 mg / mL ADC-1 protein were prepared using a 10 mM acetic acid-sodium acetate buffer system at three different pH levels: 4.9, 5.2, and 5.5. The formulations contained 60 mg / mL sucrose as a stabilizer and 0.2 mg / mL polysorbate 80 as a surfactant. The melting temperature (Tm), aggregation temperature (Tag), particle size (Radius), SEC-HPLC purity, and free drug content of the samples were measured. The stability of the three formulations was investigated under freeze-thaw conditions (-35°C and room temperature), shaking (200 rpm, 25°C), and high-temperature conditions (40°C).

[0165] 1) 10 mM acetic acid-sodium acetate, 60 mg / mL sucrose, 0.2 mg / mL polysorbate 80, pH 4.9, 20 mg / mL complex ADC-1; 2) 10 mM acetic acid-sodium acetate, 60 mg / mL sucrose, 0.2 mg / mL polysorbate 80, pH 5.2, 20 mg / mL complex ADC-1, and 3) 10 mM acetic acid-sodium acetate, 60 mg / mL sucrose, 0.2 mg / mL polysorbate 80, pH 5.5, 20 mg / mL complex ADC-1.

[0166] [Table 17] [Table 18] [Table 19-1] [Table 19-2]

[0167] Note: 2-B is a small molecule toxin, and 9-A is a small molecule compound linked to a linker. The LOD is 0.05 μg / mL, and the LOQ is 0.1 μg / mL. %PD represents the coefficient of dispersion; a smaller value indicates a more uniform particle size distribution, and so on.

[0168] Example 8. Screening of ADC-1 buffer systems Two buffer systems, 10 mM acetic acid-sodium acetate and 10 mM succinic acid-sodium succinate at pH 5.2, were selected. 60 mg / mL sucrose was used as a stabilizer, and 0.2 mg / mL polysorbate 80 was used as a surfactant. Two formulations (converted to protein concentration) containing 20 mg / mL HER3 ADC-1 were prepared. The stability of the two formulations was examined at high temperatures (40°C), 25°C, and repeated freeze-thaw cycles (-35°C and room temperature) by detecting the sample appearance, SEC-HPLC purity, R-CE purity, DAR, and free drug content.

[0169] 1) 10 mM acetic acid-sodium acetate, 60 mg / mL sucrose, 0.2 mg / mL polysorbate 80, pH 5.2, 20 mg / mL complex ADC-1, and 2) 10 mM succinic acid-sodium succinate, 60 mg / mL sucrose, 0.2 mg / mL polysorbate 80, pH 5.2, 20 mg / mL complex ADC-1.

[0170] [Table 20] [Table 21] [Table 22] [Table 23] [Table 24]

[0171] NOTE: 2-B is a small molecule toxin, 9-A is a small molecule toxin linked to a linker, "ND" stands for "not detected", the limit of detection (LOD) is 3 ppm, and the limit of quantification (LOQ) is 5 ppm.

[0172] As a result, in the acetic acid-sodium acetate buffer system and the succinic acid-sodium succinate buffer system, the samples exhibited slight opalescence and contained a small amount of particles after one week at 25°C and 40°C and repeated freeze-thawing conditions. In the succinic acid-sodium succinate buffer system, the protein particles increased after two weeks at 25°C and 40°C. Under the same conditions, there was no significant difference in the DAR value and free drug content between Formulation 1 and Formulation 2. However, at 25°C and 40°C, the SEC purity of Formulation 1 was slightly higher than that of Formulation 2. After two weeks at the high temperature of 40°C, the R-CE purity of Formulation 1 was slightly higher than that of Formulation 2. ADC-1 was found to have better stability in a 10 mM acetic acid-sodium acetate buffer system containing 60 mg / mL sucrose and 0.2 mg / mL polysorbate 80 at pH 5.2.

[0173] Example 9. Screening of ADC-1 buffer system concentrations Two buffer systems, 10 mM acetic acid-sodium acetate and 20 mM acetic acid-sodium acetate at pH 5.2, were selected. 60 mg / mL sucrose was used as a stabilizer, and 0.2 mg / mL polysorbate 80 was used as a surfactant. Two formulations (converted to protein concentration) with an ADC-1 concentration of 20 mg / mL were prepared. The stability of the two formulations under shaking (200 rpm, 25°C), repeated freeze-thawing (at -35°C and room temperature), and high temperature (40°C) conditions was investigated by detecting the sample appearance, particle size, Tm, Tagg266, SEC-HPLC purity, DAR, and free drug content.

[0174] 1) 10 mM acetic acid-sodium acetate, 60 mg / mL sucrose, 0.2 mg / mL polysorbate 80, pH 5.2, 20 mg / mL complex ADC-1, and 2) 20 mM acetic acid-sodium acetate, 60 mg / mL sucrose, 0.2 mg / mL polysorbate 80, pH 5.2, 20 mg / mL complex ADC-1.

[0175] [Table 25] [Table 26] [Table 27] [Table 28] [Table 29]

[0176] NOTE: 2-B is a small molecule toxin, and 9-A is a small molecule compound linked to a linker. The limit of detection (LOD) is 0.05 μg / mL, and the limit of quantification (LOQ) is 0.1 μg / mL.

[0177] Example 10. Confirmation of ADC-1 buffer system A 10 mM acetic acid-sodium acetate buffer system at pH 5.2 was selected to prepare a sample containing 60 mg / mL sucrose, 0.2 mg / mL polysorbate 80, and an ADC-1 concentration of 20 mg / mL. The sample's SEC-HPLC purity, R-CE purity, DAR value, free drug moiety, and binding activity were determined to examine the stability of the formulation under high temperature (40°C), 25°C, 2-8°C, -35°C, repeated freeze-thawing (-35°C and room temperature), shaking (200 rpm, 25°C), and light exposure (25°C, 5000 lx).

[0178] 10 mM acetic acid-sodium acetate, 60 mg / mL sucrose, 0.2 mg / mL polysorbate 80, pH 5.2, 20 mg / mL complex ADC-1.

[0179] [Table 30] [Table 31] [Table 32] [Table 33] [Table 34] [Table 35]

[0180] NOTE: ND stands for not detected, the limit of detection (LOD) is 3 ppm, and the limit of quantification (LOQ) is 5 ppm.

[0181] The results showed that repeated freezing and thawing, storage at -35°C, and shaking had no significant effect on the content of protein monomers and polymers, R-CE purity, DAR value, and free drug content. However, after 5 days of exposure to light, the monomer content decreased significantly, the DAR value increased to a certain extent, and the R-CE purity decreased. Therefore, care should be taken to avoid light during storage and transportation.

[0182] Example 11. Screening of ADC-1 concentration Two sets of formulations (converted to protein concentrations) were prepared using 10 mM acetic acid-sodium acetate at pH 5.2, 60 mg / mL sucrose as a stabilizer, and 0.2 mg / mL polysorbate 80 as a surfactant, with ADC-1 concentrations of 20 mg / mL and 40 mg / mL. The stability of these two sets of formulations under shaking (200 rpm, 25°C), repeated freeze-thawing (at -35°C and room temperature), and high temperature (40°C) conditions was investigated by detecting the sample appearance, particle size, Tm, Tagg266, SEC-HPLC purity, DAR, and free drug content.

[0183] 1) 10 mM acetic acid-sodium acetate, 60 mg / mL sucrose, 0.2 mg / mL polysorbate 80, pH 5.2, 20 mg / mL complex ADC-1, and 2) 10 mM acetic acid-sodium acetate, 60 mg / mL sucrose, 0.2 mg / mL polysorbate 80, pH 5.2, 40 mg / mL complex ADC-1.

[0184] [Table 36] [Table 37] [Table 38] [Table 39] [Table 40]

[0185] NOTE: 2-B is a small molecule toxin, and 9-A is a small molecule compound linked to a linker. The limit of detection (LOD) is 0.05 μg / mL, and the limit of quantification (LOQ) is 0.1 μg / mL.

[0186] Example 12. Screening of ADC-1 stabilizer and surfactant concentrations Two sets of formulation samples were prepared using a 10 mM acetic acid-sodium acetate buffer system at pH 5.2, containing 60 mg / mL or 120 mg / mL sucrose, 0.2 mg / mL or 0.4 mg / mL polysorbate 80, and an ADC-1 concentration of 20 mg / mL. After filtering through a 0.22 μm filter, 5.3 mL of each was filled into a 20 mL vial, half of which was filled with a plug, and freeze-dried according to the freeze-drying process parameters in the table below. The entire vial was then filled with a plug and capped. The appearance, moisture, reconstitution time, SEC purity, R-CE purity, DAR value, and free drug content were examined to compare the changes in product quality before and after freeze-drying of the same formulation. The impact of the freeze-drying process on product quality was evaluated, and the quality of different formulations was compared to screen the stabilizer and surfactant concentrations.

[0187] 1) 10 mM acetic acid-sodium acetate, 60 mg / mL sucrose, 0.2 mg / mL polysorbate 80, pH 5.2, 20 mg / mL ADC-1, and 2) 10 mM acetic acid-sodium acetate, 120 mg / mL sucrose, 0.4 mg / mL polysorbate 80, pH 5.2, 20 mg / mL ADC-1.

[0188] [Table 41] [Table 42-1] [Table 42-2]

[0189] [Table 43]

[0190] Note: For formulation 1, the sample before freeze-drying is labeled DS-1 and the sample after freeze-drying is labeled DP-1. For formulation 2, the sample before freeze-drying is labeled DS-2 and the sample after freeze-drying is labeled DP-2.

[0191] As a result, the appearance of both the pre- and post-lyophilization samples of the two formulations was acceptable. However, compared with the pre-lyophilization sample, the post-lyophilization sample of the same formulation had a lower free drug content, while the monomer content, polymer content, DAR value, and R-CE purity all remained significantly unchanged, indicating that the lyophilization process had no significant impact on product quality. There was no significant difference in any of the detection items of the lyophilized products of the two formulations, so the sucrose concentration was set at 60-120 mg / mL and the polysorbate 80 concentration at 0.2-0.4 mg / mL.

[0192] Example 13. Confirmation of stabilizer concentration and surfactant concentration of ADC-1 1 A 10 mM acetic acid-sodium acetate buffer system at pH 5.2 was selected to prepare a formulation sample containing 60 mg / mL sucrose, 0.2 mg / mL polysorbate 80, and an ADC-1 concentration of 20 mg / mL. After filtering through a 0.22 μm filter, 5.3 mL of the sample was filled into a 20 mL vial, half of which was filled with a plug, and freeze-dried according to the freeze-drying process parameters in the table below. The entire vial was filled with a plug, and the vial was capped. The stabilizer concentration and surfactant concentration were confirmed by detecting the appearance, moisture content, reconstitution time, SEC purity, R-CE purity, DAR value, and free drug content.

[0193] [Table 44] [Table 45]

[0194] [Table 46]

[0195] Note: For Formulation 1, the sample before freeze-drying was labeled DS-1, and the sample after freeze-drying was labeled DP-1.

[0196] As a result, the pre-freezing time was extended from 120 minutes to 180 minutes, the secondary drying time was extended from 260 minutes to 480 minutes, and the secondary drying vacuum was adjusted from 1 Pa to 5 Pa. Compared with the sample before freeze-drying, the free drug content, monomer content, polymer content, DAR value and R-CE purity of the freeze-dried sample all showed no significant changes, indicating that this freeze-drying process had no obvious impact on product quality and that the formulation had good stability.

[0197] Example 14. Confirmation of stabilizer concentration and surfactant concentration of ADC-1 2 A 10 mM acetic acid-sodium acetate buffer system at pH 5.2 was selected to prepare a formulation sample containing 120 mg / mL of sucrose, 0.4 mg / mL of polysorbate 80, and an ADC-1 concentration of 20 mg / mL. After filtering through a 0.22 μm filter, 5.3 mL of the formulation was filled into a 20 mL vial, half of which was filled with a plug, and freeze-dried according to the freeze-drying process parameters in the table below. The entire vial was filled with a plug, and the vial was capped. The stabilizer concentration and surfactant concentration were confirmed by detecting the appearance, moisture content, reconstitution time, SEC purity, R-CE purity, DAR value, and free drug content.

[0198] [Table 47] [Table 48]

[0199] [Table 49]

[0200] Note: For Formulation 1, the sample before freeze-drying was labeled DS-1, and the sample after freeze-drying was labeled DP-1.

[0201] As a result, it was found that when the primary drying temperature was increased from -20°C to -15°C, the primary drying time was shortened from 2400 minutes to 2100 minutes, and the secondary drying time was extended from 240 minutes to 480 minutes, the free drug content of the freeze-dried sample decreased compared to the sample before freeze-drying, while the monomer content, polymer content, DAR value, and R-CE purity all remained significantly unchanged, indicating that this freeze-drying process had no obvious effect on product quality and that the formulation had good stability.

[0202] Example 15. Formulation of Selectable Anti-HER3 Antibodies The present disclosure provides a formulation of "18-22 mg / mL anti-HER3 antibody HER3-29 agent, 60-90 mg / mL sucrose, 10 mM acetic acid-sodium acetate, pH 5.0-5.4" 1) 20 mg / mL anti-HER3 antibody, 60 mg / mL sucrose, 10 mM acetic acid-sodium acetate pH 5.0, 2) 20 mg / mL anti-HER3 antibody, 60 mg / mL sucrose, 10 mM acetic acid-sodium acetate pH 5.2, 3) 20 mg / mL anti-HER3 antibody, 60 mg / mL sucrose, 10 mM acetic acid-sodium acetate pH 5.4, 4) 20 mg / mL anti-HER3 antibody, 90 mg / mL sucrose, 10 mM acetic acid-sodium acetate pH 5.0, 5) 20 mg / mL anti-HER3 antibody, 90 mg / mL sucrose, 10 mM acetic acid-sodium acetate pH 5.2, and 6) 20 mg / mL anti-HER3 antibody, 90 mg / mL sucrose, 10 mM acetic acid-sodium acetate pH 5.4, Including, but not limited to:

[0203] Example 16. Formulation methods for selective anti-HER3 antibody-drug conjugates The present disclosure provides a formulation method for "18 to 22 mg / mL anti-HER3 antibody-drug conjugate, 60 to 120 mg / mL sucrose, 0.2 to 0.4 mg / mL polysorbate 80, 10 mM acetic acid-sodium acetate, pH 5.0 to 5.4," 1) 20 mg / mL ADC-1, 60 mg / mL sucrose, 0.2 mg / mL polysorbate 80, 10 mM acetic acid-sodium acetate pH 5.0, 2) 20 mg / mL ADC-1, 60 mg / mL sucrose, 0.2 mg / mL polysorbate 80, 10 mM acetic acid-sodium acetate pH 5.2, 3) 20 mg / mL ADC-1, 60 mg / mL sucrose, 0.2 mg / mL polysorbate 80, 10 mM acetic acid-sodium acetate pH 5.4, 4) 20 mg / mL ADC-1, 120 mg / mL sucrose, 0.2 mg / mL polysorbate 80, 10 mM acetic acid-sodium acetate pH 5.0, 5) 20 mg / mL ADC-1, 120 mg / mL sucrose, 0.2 mg / mL polysorbate 80, 10 mM acetic acid-sodium acetate pH 5.2, 6) 20 mg / mL ADC-1, 120 mg / mL sucrose, 0.2 mg / mL polysorbate 80, 10 mM acetic acid-sodium acetate pH 5.4, 7) 20 mg / mL ADC-1, 60 mg / mL sucrose, 0.4 mg / mL polysorbate 80, 10 mM acetic acid-sodium acetate pH 5.0, 8) 20 mg / mL ADC-1, 60 mg / mL sucrose, 0.4 mg / mL polysorbate 80, 10 mM acetic acid-sodium acetate pH 5.2, 9) 20 mg / mL ADC-1, 60 mg / mL sucrose, 0.4 mg / mL polysorbate 80, 10 mM acetic acid-sodium acetate pH 5.4, 10) 20 mg / mL ADC-1, 120 mg / mL sucrose, 0.4 mg / mL polysorbate 80, 10 mM acetic acid-sodium acetate pH 5.0, 11) 20 mg / mL ADC-1, 120 mg / mL sucrose, 0.4 mg / mL polysorbate 80, 10 mM acetic acid-sodium acetate pH 5.2, and 12) 20 mg / mL ADC-1, 120 mg / mL sucrose, 0.4 mg / mL polysorbate 80, 10 mM acetic acid-sodium acetate pH 5.4, Including, but not limited to:

Claims

1. 1. A pharmaceutical composition comprising an anti-HER3 antibody-drug conjugate having the structure shown in the formula: 【Chemistry 1】 Among them, n is 1 to 8; Pc is an anti-HER3 antibody comprising a heavy chain variable region and a light chain variable region, the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 shown in SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 shown in SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6, respectively; further comprising an acetate buffer, preferably an acetic acid-sodium acetate buffer, having a pH of 4.9 to 5.5, preferably a pH of about 5.2; Pharmaceutical compositions.

2. The anti-HER3 antibody comprising a heavy chain variable region represented by SEQ ID NO: 7 and a light chain variable region represented by SEQ ID NO: 8; Preferably, the anti-HER3 antibody is A heavy chain represented by SEQ ID NO: 9 and a light chain represented by SEQ ID NO:

10. The pharmaceutical composition of claim 1.

3. The n is 3 to 5, and preferably n is about 4. The pharmaceutical composition according to claim 1 or 2.

4. The pharmaceutical composition further comprises a surfactant, preferably a polysorbate, more preferably polysorbate 80 or polysorbate 20, most preferably polysorbate 80. The pharmaceutical composition according to any one of claims 1 to 3.

5. The concentration of the surfactant is 0.05 to 0.6 mg / mL, preferably 0.1 to 0.4 mg / mL, more preferably about 0.2 mg / mL; The pharmaceutical composition according to claim 4.

6. The composition further comprises a sugar, preferably the sugar is selected from sucrose and α,α-trehalose dihydrate, more preferably sucrose. The pharmaceutical composition according to any one of claims 1 to 5.

7. the sugar concentration is 10-150 mg / mL, preferably 40-120 mg / mL, more preferably 40-80 mg / mL, and most preferably about 60 mg / mL; The pharmaceutical composition according to claim 6.

8. The concentration of the buffer is 1 to 50 mM, preferably 5 to 15 mM, more preferably about 10 mM; The pharmaceutical composition according to any one of claims 1 to 7.

9. the concentration of the antibody-drug conjugate is 1 to 40 mg / mL, preferably 10 to 30 mg / mL, more preferably 15 to 25 mg / mL, and most preferably about 20 mg / mL; The pharmaceutical composition according to any one of claims 1 to 8.

10. A pharmaceutical composition according to any one of claims 1 to 9, comprising the following ingredients: (a) 1 to 40 mg / mL of the anti-HER3 antibody-drug conjugate, (b) 0.05 to 0.6 mg / mL of polysorbate, (c) 10 to 150 mg / mL of sugar, and (d) 1 to 50 mM of acetate buffer, wherein the pH of the pharmaceutical composition is 4.9 to 5.5; Preferably, said pharmaceutical composition comprises the following ingredients: (a) 10 to 30 mg / mL of the anti-HER3 antibody-drug conjugate, (b) 0.1 to 0.4 mg / mL of polysorbate, (c) 40 to 120 mg / mL of sugar, and (d) 5 to 15 mM of acetate buffer, wherein the pH of the pharmaceutical composition is 5.0 to 5.5; More preferably, said pharmaceutical composition comprises the following ingredients: (a) 15 to 25 mg / mL of the anti-HER3 antibody-drug conjugate, (b) 0.1 to 0.4 mg / mL of polysorbate 80, (c) 40 to 80 mg / mL of sucrose, and (d) 5 to 15 mM of acetic acid-sodium acetate buffer, wherein the pH of the pharmaceutical composition is 5.1 to 5.3; Most preferably, the pharmaceutical composition comprises the following ingredients: (a) about 20 mg / mL of the anti-HER3 antibody-drug conjugate, (b) about 0.2 mg / mL of polysorbate 80, (c) about 60 mg / mL of sucrose, and (d) about 10 mM of acetic acid-sodium acetate buffer, wherein the pH of the pharmaceutical composition is about 5.

2. The pharmaceutical composition according to any one of claims 1 to 9.

11. A lyophilized formulation comprising an anti-HER3 antibody-drug conjugate, characterized in that after reconstitution it can form the pharmaceutical composition according to any one of claims 1 to 10. Lyophilized formulation.

12. A method for preparing a lyophilized formulation comprising an anti-HER3 antibody-drug conjugate, the method comprising the step of lyophilizing the pharmaceutical composition according to any one of claims 1 to 10. method.

13. A method for treating a disease, comprising administering to a patient an effective amount of the pharmaceutical composition according to any one of claims 1 to 10 or the lyophilized formulation according to claim 11; Preferably, the disease is a tumor or cancer, more preferably, the disease is selected from breast cancer, non-small cell lung cancer, gastric cancer, ovarian cancer, prostate cancer, bladder cancer, colorectal cancer, head and neck squamous cell carcinoma and melanoma. method.

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