Antibody formulation

Abstract

The present disclosure relates to the field of therapeutic antibodies, and specifically, to a pharmaceutical composition comprising an antibody, a buffer solution, and an additional excipient. The pharmaceutical composition can maintain the stability of the antibody after long-term storage and can be directly infused, thereby facilitating clinical use. In the pharmaceutical composition, the antibody component is present at a high concentration, thus featuring convenience of storage and transport and cost-efficiency. In addition, the present disclosure further relates to use of the pharmaceutical composition in preparing a medicament for the prevention and / or treatment and / or adjuvant therapy and / or diagnosis of a tumor.

Description

Antibody preparations

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application is based on and claims priority to CN application No. 2023116187657, filed on November 29, 2023, and CN application No. 2024116719780, filed on November 21, 2024. The contents of the CN application are hereby incorporated into this application as a whole. Technical Field

[0003] The present invention relates to the field of therapeutic antibodies, and more specifically, to an anti-EGFR antibody pharmaceutical composition and its medical use. Background Art

[0004] Protein-based pharmaceutical preparations, particularly antibodies, are available for multiple administration routes, including intravenous, intramuscular, and subcutaneous injection. However, due to the inherent properties of protein molecules, liquid preparations often face stability issues and require stringent storage conditions. Furthermore, protein preparations are prone to aggregates or particulates during storage, which can affect the biological, chemical, and physical stability of the preparation.

[0005] Recombinant anti-epidermal growth factor receptor (EGFR) human-mouse chimeric monoclonal antibody has a complex higher-order structure and poor intrinsic stability. Existing formulations of this antibody are prone to forming aggregates and protein particles, requiring in-line filtration using a 0.22μm filter before injection.

[0006] Therefore, for anti-EGFR antibodies, there is a need in the art for antibody preparations with good stability and ease of use. Summary of the Invention

[0007] After extensive experiments and repeated exploration, the inventors of the present application unexpectedly discovered that an antibody pharmaceutical composition with a specific composition has excellent stability, contains a high concentration of protein active ingredients and / or does not require dilution or filtration before use. Therefore, the present application relates to a pharmaceutical composition that is particularly advantageous for long-term storage and clinical use of drugs, as well as its preparation method and use.

[0008] In a first aspect, the present application relates to a pharmaceutical composition comprising 0.5-10 g / L of an antibody, 6-12 g / L of a stabilizer, 0.01-0.5 g / L of a surfactant, and a buffer system; the pH of the liquid preparation is 6.0-8.0;

[0009] The stabilizer is selected from one or more of sodium chloride, glycine, sucrose, sorbitol, and proline;

[0010] The surfactant is selected from polysorbate 20 or polysorbate 80;

[0011] The buffer system is selected from a phosphate buffer system, a citrate buffer system or a histidine buffer system.

[0012] In some embodiments, the pharmaceutical composition comprises 1 to 8 g / L of the antibody, preferably 1 to 3 g / L or 4 to 6 g / L of the antibody.

[0013] In some embodiments, the pharmaceutical composition comprises 0.5-5 g / L of antibody, 6-12 g / L of stabilizer, 0.01-0.5 g / L of surfactant, and a buffer system; the pH of the liquid preparation is 6.0-8.0;

[0014] The stabilizer is selected from one or more of sodium chloride, glycine, sucrose, sorbitol, and proline;

[0015] The surfactant is selected from polysorbate 20 or polysorbate 80;

[0016] The buffer system is selected from a phosphate buffer system, a citrate buffer system or a histidine buffer system.

[0017] In certain embodiments, the pharmaceutical composition comprises 1.8-2.2 g / L of antibody, 7.632-9.328 g / L of stabilizer, 0.05-0.15 g / L of surfactant, and a buffer system; the pH of the pharmaceutical composition is 6.4-7.0.

[0018] In certain embodiments, the pharmaceutical composition comprises 4-6 g / L of antibody, 7-10 g / L of stabilizer, 0.05-0.15 g / L of surfactant, and a buffer system; the pH of the pharmaceutical composition is 6.4-7.0.

[0019] In certain embodiments, the pharmaceutical composition comprises, based on 1000 ml, 1.8-2.2 g / L (e.g., 1.9 g / L, 2.0 g / L, or 2.1 g / L) of antibody, 7.632-9.328 g / L (0.13 mol / L-0.16 mol / L, e.g., 0.14 mol / L or 0.15 mol / L) of NaCl, polysorbate 80, 0.05-0.15 g / L (e.g., 0.06 g / L, 0.07 g / L, 0.08 g / L, 0.09 g / L, 0.10 g / L, 0.11 g / L, 0.12 g / L, 0.13 g / L, or 0.14 g / L), sodium dihydrogen phosphate 0.369-0.451 g / L (2.7 mmol / L-3.3 mmol / L, e.g., 2.8 mmol / L, 2.9 mmol / L, 3.0 mmol / L, 3.1 mmol / L, or 3.2 mmol / L), disodium hydrogen phosphate 0.9-1 .1g / L (6.4mmol / L~7.7mmol / L, for example, 6.5mmol / L, 6.6mmol / L, 6.7mmol / L, 6.8mmol / L, 6.9mmol / L, 7.0mmol / L, 7.1mmol / L, 7.2mmol / L, 7.3mmol / L, 7.4mmol / L, 7.5mmol / L or 7.6mmol / L); the pH of the pharmaceutical composition is 6.4~7.0 (for example, 6.5, 6.6, 6.7, 6.8 or 6.9).

[0020] In certain embodiments, the pharmaceutical composition comprises, based on 1000 ml, 4 to 6 g / L (e.g., 4.0 g / L, 4.5 g / L, 5.0 g / L, 5.5 g / L, or 6.0 g / L) of the antibody, 7.632 to 9.328 g / L (0.13 mol / L to 0.16 mol / L, e.g., 0.14 mol / L or 0.15 mol / L) of NaCl, polysorbate 80, 0.05-0.15 g / L (e.g., 0.06 g / L, 0.07 g / L, 0.08 g / L, 0.09 g / L, 0.10 g / L, 0.11 g / L, 0.12 g / L, 0.13 g / L, or 0.14 g / L), sodium dihydrogen phosphate 0.369-0.451 g / L (2.7 mmol / L-3.3 mmol / L, e.g., 2.8 mmol / L, 2.9 mmol / L, 3.0 mmol / L, 3.1 mmol / L, or 3.2 mmol / L), disodium hydrogen phosphate 0.9-1 .1g / L (6.4mmol / L~7.7mmol / L, for example, 6.5mmol / L, 6.6mmol / L, 6.7mmol / L, 6.8mmol / L, 6.9mmol / L, 7.0mmol / L, 7.1mmol / L, 7.2mmol / L, 7.3mmol / L, 7.4mmol / L, 7.5mmol / L or 7.6mmol / L); the pH of the pharmaceutical composition is 6.4~7.0 (for example, 6.5, 6.6, 6.7, 6.8 or 6.9).

[0021] In certain embodiments, the sodium dihydrogen phosphate is a monohydrate.

[0022] In certain embodiments, the sodium phosphate dibasic is anhydrous.

[0023] In certain embodiments, the antibody is an anti-EGFR antibody.

[0024] In certain embodiments, the anti-EGFR antibody comprises heavy chain CDRs as set forth in SEQ ID NOs: 1-3, respectively, and light chain CDRs as set forth in SEQ ID NOs: 4-6, respectively. In certain embodiments, the anti-EGFR antibody has a heavy chain variable region as set forth in SEQ ID NO: 7 and a light chain variable region as set forth in SEQ ID NO: 8. In certain embodiments, the anti-EGFR antibody has a heavy chain as set forth in SEQ ID NO: 9 and a light chain as set forth in SEQ ID NO: 10.

[0025] In certain embodiments, based on 1000 mL, the pharmaceutical composition comprises 2.0 g of an anti-EGFR antibody, 8.48 g of sodium chloride, 0.41 g of sodium dihydrogen phosphate (monohydrate), 1.0 g of anhydrous disodium hydrogen phosphate, and 0.1 g of polysorbate 80; the pH of the liquid preparation is 6.7.

[0026] In certain embodiments, based on 1000 mL, the pharmaceutical composition comprises 5.0 g of anti-EGFR antibody, 8.48 g of sodium chloride, 0.41 g of sodium dihydrogen phosphate (monohydrate), 1.0 g of anhydrous disodium hydrogen phosphate, and 0.1 g of polysorbate 80; the pH of the liquid preparation is 6.7.

[0027] In a second aspect, the present application relates to another pharmaceutical composition comprising 10-100 g / L of an antibody, 50-100 g / L of a stabilizer, 0.05-1.0 g / L of a surfactant, and a buffer system; the pH of the liquid preparation is 5.0-7.5;

[0028] The stabilizer is selected from one or more of sodium chloride, glycine, sucrose, sorbitol, and proline;

[0029] The surfactant is selected from polysorbate 20 or polysorbate 80;

[0030] The buffer system is selected from a phosphate buffer system, a citrate buffer system or a histidine buffer system.

[0031] In certain embodiments, the pharmaceutical composition comprises 20-60 g / L of antibody, 0.1-0.3 g / L of surfactant, and 80-90 g / L of stabilizer, and the pH of the pharmaceutical composition is 5.2-6.2.

[0032] In certain embodiments, the pharmaceutical composition comprises 20 to 60 g / L (e.g., 40 g / L) of the antibody, 10 mmol / L to 20 mmol / L (e.g., 11 mmol / L, 12 mmol / L, 13 mmol / L, 14 mmol / L, 15 mmol / L, 16 mmol / L, 17 mmol / L, 18 mmol / L, or 19 mmol / L) of histidine and histidine hydrochloride per 1000 ml, and polysorbate 80 is 0.1-0.3 g / L (e.g., 0.15 g / L, 0.2 g / L or 0.25 g / L), sucrose 80 g / L-90 g / L (e.g., 81 g / L, 82 g / L, 83 g / L, 84 g / L, 85 g / L, 86 g / L, 87 g / L, 88 g / L or 89 g / L), and the pH of the pharmaceutical composition is 5.2-6.2 (e.g., 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0 or 6.1).

[0033] In certain embodiments, the antibody is an anti-EGFR antibody.

[0034] In certain embodiments, the anti-EGFR antibody comprises heavy chain CDRs as set forth in SEQ ID NOs: 1-3, respectively, and light chain CDRs as set forth in SEQ ID NOs: 4-6, respectively. In certain embodiments, the anti-EGFR antibody has a heavy chain variable region as set forth in SEQ ID NO: 7 and a light chain variable region as set forth in SEQ ID NO: 8. In certain embodiments, the anti-EGFR antibody has a heavy chain as set forth in SEQ ID NO: 9 and a light chain as set forth in SEQ ID NO: 10.

[0035] In certain embodiments, the pharmaceutical composition comprises 20 g / L anti-EGFR antibody, 10-20 mM histidine-histidine hydrochloride, 0.2 g / L polysorbate 80, and 80 g / L sucrose per 1000 ml, and the pH of the pharmaceutical composition is 5.5±0.3.

[0036] In certain embodiments, the pharmaceutical composition comprises 20 g / L anti-EGFR antibody, 20 mM histidine-histidine hydrochloride, 0.2 g / L polysorbate 80, and 80 g / L sucrose per 1000 ml, and the pH of the pharmaceutical composition is 5.5±0.3.

[0037] In certain embodiments, the pharmaceutical composition comprises 20 g / L of anti-EGFR antibody, 0.67 g / L of histidine, 3.29 g / L of histidine hydrochloride, 0.2 g / L of polysorbate 80, and 80 g / L of sucrose per 1000 ml, and the pH of the pharmaceutical composition is 5.5±0.3.

[0038] In certain embodiments, the pharmaceutical composition comprises 20-60 g / L of anti-EGFR antibody, 90 g / L of sucrose, 0.1-0.2 g / L of polysorbate 80, and 20 mM of histidine-histidine hydrochloride per 1000 ml, and the pH of the pharmaceutical composition is 5.5±0.3.

[0039] In certain embodiments, the pharmaceutical composition comprises 20 g / L anti-EGFR antibody, 90 g / L sucrose, 0.1 g / L polysorbate 80, and 20 mM histidine-histidine hydrochloride per 1000 ml, and the pH of the pharmaceutical composition is 5.5±0.3.

[0040] In certain embodiments, the pharmaceutical composition comprises 20-60 g / L of anti-EGFR antibody, 10 mM histidine-histidine hydrochloride, 0.2 g / L of polysorbate 80, and 80 g / L of sucrose per 1000 ml, and the pH of the pharmaceutical composition is 5.5±0.3.

[0041] In certain embodiments, the pharmaceutical composition comprises 60 g / L of anti-EGFR antibody, 10 mM histidine-histidine hydrochloride, 0.2 g / L polysorbate 80, and 80 g / L sucrose per 1000 ml, and the pH of the pharmaceutical composition is 5.2-5.8.

[0042] In certain preferred embodiments, the pharmaceutical compositions of the present invention are stable at 5°C ± 3°C and can be stored for at least about 2 weeks, at least about 1 month, at least about 2 months, at least about 3 months, at least about 6 months, at least about 9 months, at least about 1 year, or at least about 2 years. In certain preferred embodiments, the pharmaceutical compositions of the present invention are stable at about 40°C and can be stored for at least about 2 weeks, at least about 1 month, at least about 2 months, at least about 3 months, at least about 6 months, at least about 9 months, at least about 1 year, or at least about 2 years. In certain preferred embodiments, the pharmaceutical compositions of the present invention are stable at room temperature (e.g., about 25°C) and can be stored for at least about 2 weeks, at least about 1 month, at least about 2 months, at least about 3 months, at least about 6 months, at least about 9 months, at least about 1 year, or at least about 2 years.

[0043] In certain embodiments, the strength of the pharmaceutical composition is 150 mg (10 ml) / bottle to 2000 mg (10 ml) / bottle.

[0044] In certain embodiments, the strength of the pharmaceutical composition is selected from the following group: 200 mg (10 ml) / bottle, 600 mg (10 ml) / bottle, 900 mg (10 ml) / bottle, 1200 mg (10 ml) / bottle or 1500 mg (10 ml) / bottle.

[0045] The pharmaceutical composition of the present invention can be prepared by combining various components at predetermined concentrations using methods known in the art. For example, the antibody can be dialyzed into a solution containing the other components of the liquid formulation and adjusted to the desired concentration. The formulation is sterilized by filtration using a filter with a pore size of 0.22 μm. The prepared pharmaceutical composition is packaged for ease of use. The packaging material can be a glass bottle (e.g., a vial), a metal alloy container, a prefilled syringe, or a pen injector.

[0046] In certain embodiments, the present invention provides a method for stabilizing an antibody, characterized in that the method is performed by combining 0.5-5 g / L of an antibody, 6-12 g / L of a stabilizer, 0.01-0.5 g / L of a surfactant, and a buffer system, and controlling the pH of the combination to be 6.0-8.0.

[0047] In certain embodiments, the present invention provides a method for stabilizing an antibody, characterized in that the method is performed by combining 10-100 g / L of an antibody, 50-100 g / L of a stabilizer, 0.05-1.0 g / L of a surfactant, and a buffer system, and controlling the pH of the combination to be 5.0-7.5.

[0048] In certain embodiments, the antibody, surfactant, stabilizer, surfactant and / or buffer system are as described above.

[0049] The pharmaceutical composition of the present invention may be contained in any container suitable for storing drugs and other pharmaceutical compositions. For example, the pharmaceutical composition of the present invention may be contained in a sealed and sterilized plastic container, metal alloy container or glass container with a certain volume, such as an ampoule, a vial or a syringe. Therefore, in certain preferred embodiments, the pharmaceutical composition of the present invention is contained in a glass vial (e.g., an ampoule or a vial), a syringe or a microinfusion device. In certain exemplary embodiments, the pharmaceutical composition of the present invention is contained in a glass vial (e.g., an ampoule or a vial).

[0050] In certain embodiments, the pharmaceutical composition is a liquid.

[0051] In a third aspect, the present invention further provides a solid preparation prepared by freeze-drying the pharmaceutical composition described above. In certain embodiments, the solid preparation is a freeze-dried powder for injection.

[0052] In a fourth aspect, the present invention further provides an article comprising a container containing the pharmaceutical composition of the present invention. In certain embodiments, the container is a glass bottle, a metal alloy container, or a prefilled syringe. In certain embodiments, the article is a liquid or solid preparation for injection.

[0053] In a fifth aspect, the present application relates to a pharmaceutical preparation, which is obtained by diluting the pharmaceutical composition of the second aspect of the present invention with a hydrochloric acid-histidine buffer. In certain embodiments, the dilution multiple is 5-50 times. In certain embodiments, the dilution multiple is 10-30 times. In certain embodiments, the dilution multiple is 10 times. In certain embodiments, the pH of the hydrochloric acid-histidine buffer is 5.2 to 6.2 (e.g., 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0 or 6.1). In certain embodiments, the pharmaceutical preparation is a liquid preparation for injection.

[0054] In a sixth aspect, the present application relates to the use of any of the above pharmaceutical compositions, pharmaceutical preparations or products in the preparation of a drug, wherein the drug is used for:

[0055] (a) Prevention and / or treatment of tumors;

[0056] (b) Adjuvant treatment of tumors;

[0057] (c) Diagnosis of tumors;

[0058] (d). Any combination of (a)-(c).

[0059] In certain embodiments, the drug is administered separately, in combination, simultaneously, or sequentially with another pharmaceutically active agent; optionally, the other pharmaceutically active agent is a chemotherapeutic drug.

[0060] In certain embodiments, the present application provides any of the above pharmaceutical compositions, pharmaceutical preparations or products for use in:

[0061] (a) Prevention and / or treatment of tumors;

[0062] (b) Adjuvant treatment of tumors;

[0063] (c) Diagnosis of tumors;

[0064] (d). Any combination of (a)-(c).

[0065] In certain embodiments, the present application provides a method for preventing, treating, adjuvant treating and / or diagnosing tumors, comprising administering to an individual in need thereof a therapeutically effective amount of any one of the pharmaceutical compositions, pharmaceutical preparations or products described above.

[0066] In certain embodiments, the tumor is selected from breast cancer, lung cancer, such as non-small cell lung cancer or squamous cell lung cancer, liver cancer, stomach cancer, intestinal cancer, such as colon cancer or rectal cancer, esophageal cancer, ovarian cancer, cervical cancer, kidney cancer, prostate cancer, bladder cancer, pancreatic cancer, Merkel cell carcinoma, bile duct cancer, nasopharyngeal carcinoma, head and neck squamous cell carcinoma, glioma, melanoma, leukemia and lymphoma.

[0067] In certain embodiments, the tumor is selected from colorectal cancer; preferably, the tumor is selected from RAS wild-type metastatic colorectal cancer.

[0068] In certain embodiments, the area under the drug-time curve (AUC) of any of the above pharmaceutical compositions, pharmaceutical preparations or products in a subject is 5000 h*μg / mL or more, 6000 h*μg / mL or more, 7000 h*μg / mL or more, 8000 h*μg / mL or more, 9000 h*μg / mL or more, 10000 h*μg / mL or more, or 11000 h*μg / mL or more, and in certain embodiments, no more than 14000 h*μg / mL.

[0069] In certain embodiments, the AUC is AUC0-t .

[0070] In certain embodiments, the peak concentration (C max ) is 50~500μg / mL, 60~450μg / mL, 70~400μg / mL, 80~300μg / mL, 90~250μg / mL, or 100~200μg / mL.

[0071] In certain embodiments, the terminal elimination half-life (t 1 / 2 ) is 10 hours or more, 20 hours or more, 30 hours or more, 40 hours or more, or 45 hours or more, and in certain embodiments, not more than 60 hours.

[0072] In certain embodiments, the clearance rate (CL) of any of the above-mentioned pharmaceutical compositions, pharmaceutical preparations or products in the subject is less than 0.1 L / h, less than 0.09 L / h, less than 0.08 L / h, less than 0.07 L / h, less than 0.06 L / h, less than 0.05 L / h, and in certain embodiments, not less than 0.03 L / h.

[0073] In certain embodiments, the dosage administered to the subject is 50 to 500 mg / m2 calculated based on body surface area (BSA). 2 , preferably 100 to 400 mg / m 2 More preferably, 250 mg / m 2 .

[0074] In certain embodiments, the number of administrations to the subject in one administration cycle is 1 to 5 times (eg, 1, 2, 3, 4, or 5 times), preferably 1 time.

[0075] In certain embodiments, the administration to the subject is oral or intravenous, preferably intravenous.

[0076] In a seventh aspect, the present application relates to the use of any of the above-mentioned pharmaceutical compositions, pharmaceutical preparations or products in the preparation of liquid preparations for injection.

[0077] definition

[0078] Unless otherwise indicated, scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. Furthermore, the cell culture, biochemistry, nucleic acid chemistry, immunology, and other laboratory procedures used herein are conventional procedures widely used in the relevant fields. To facilitate a better understanding of the present invention, definitions and explanations of relevant terms are provided below.

[0079] As used herein, the term "antibody" is used in the broadest sense and includes various antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies) and antibody fragments, as long as they exhibit the desired antigen-binding activity. For example, an immunoglobulin molecule can be composed of two pairs of polypeptide chains, each pair having one light chain (LC) and one heavy chain (HC). Antibody light chains can be classified as kappa (κ) and lambda (λ) light chains. Heavy chains can be classified as μ, δ, γ, α or ε, and define the isotype of the antibody as IgM, IgD, IgG, IgA and IgE, respectively. Within the light and heavy chains, the variable and constant regions are connected by a "J" region of about 12 or more amino acids, and the heavy chain also contains a "D" region of about 3 or more amino acids. Each heavy chain consists of a heavy chain variable region (VH) and a heavy chain constant region (CH). The heavy chain constant region consists of three domains (CH1, CH2 and CH3). Each light chain is composed of a light chain variable region (VL) and a light chain constant region (CL). The light chain constant region consists of a single domain, CL. The constant domain is not directly involved in antibody-antigen binding but exhibits various effector functions, such as mediating the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system. The VH and VL regions can be further subdivided into highly variable regions, known as complementarity-determining regions (CDRs), interspersed with more conserved regions known as framework regions (FRs). Each VH and VL consists of three CDRs and four FRs, arranged from amino-terminus to carboxyl-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions (VH and VL) of each heavy / light chain pair form the antigen-binding site. The distribution of amino acids among regions or domains can follow the definitions of Kabat, Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987 and 1991)), or Chothia & Lesk (1987) J. Mol. Biol. 196:901-917; Chothia et al. (1989) Nature 342:878-883.

[0080] Herein, unless the context clearly indicates otherwise, when referring to the term "antibody", it includes not only intact antibodies, but also antigen-binding fragments of antibodies.

[0081] As used herein, the term "complementarity determining region" or "CDR" refers to the amino acid residues in the variable region of an antibody that are responsible for antigen binding. The precise boundaries of these amino acid residues can be defined according to various numbering systems known in the art, such as the AbM numbering system (Martin ACR, Cheetham JC, Rees AR (1989) Modelling antibody hypervariable loops: A combined algorithm. Proc Natl Acad Sci USA 86: 9268-9272) or the IMGT numbering system (Lefranc et al., Dev. Comparat. Immunol. 27: 55-77, 2003). For a given antibody, those skilled in the art will readily identify the CDRs defined by each numbering system. Furthermore, the correspondence between different numbering systems is well known to those skilled in the art (e.g., see Lefranc et al., Dev. Comparat. Immunol. 27: 55-77, 2003).

[0082] In the present invention, the CDRs contained in the antibodies or antigen-binding fragments thereof of the present invention can be identified according to various numbering systems known in the art. In certain embodiments, the CDRs contained in the antibodies or antigen-binding fragments thereof of the present invention are preferably identified by the IMGT, Kabat, Chothia or AbM numbering systems.

[0083] The term "anti-EGFR antibody" refers to an antibody that can specifically bind to EGFR. The term "specific binding" refers to the formation of a relatively stable complex between the antibody and the antigen under physiological conditions. Specific binding can be achieved by approximately 1×10 -9 Methods for determining whether two molecules specifically bind are well known in the art, such as bioluminescence interferometry or surface plasmon resonance technology (eg, Biacore).

[0084] As used herein, the term "liquid preparation" refers to a combination of at least one active ingredient (e.g., an antibody capable of producing a biologically active substance in a subject) and at least one inactive ingredient, wherein the inactive ingredient is suitable for administration to the subject when combined with the active ingredient. Typically, the liquid preparation does not contain any ingredients that would cause unacceptable toxicity to the subject to whom the preparation is administered. In this article, the subject may include a mammal, preferably a human.

[0085] As used herein, the "stability" of an antibody refers to that an antibody substantially retains its physical stability and / or chemical stability and / or biological activity after storage. The storage period is generally selected based on the predetermined shelf life of the formulation. Analytical techniques for measuring antibody stability are well known in the art and stability can be determined at a selected temperature and for a selected time (e.g., antibody stability during storage at 40°C for at least 28 days and testing, or antibody stability during storage at 5°C ± 3°C for at least 12 months and testing).

[0086] With respect to physical stability, an antibody in a formulation is considered to have retained its physical stability if it exhibits substantially no evidence of aggregation, precipitation and / or denaturation as determined by visual inspection of color and / or clarity, or by UV light scattering or by size exclusion chromatography.

[0087] For chemical stability, it can be assessed by detecting and quantitatively modifying the antibody. Chemical modification may involve size changes (e.g., shearing), such as can be assessed by using size exclusion chromatography, SDS-PAGE and / or matrix-assisted laser desorption ionization / time of flight mass spectrometry (MALDI / TOF MS). Other types of chemical modifications include charge changes (e.g., changes caused by deamidation), such as can be assessed by ion exchange chromatography.

[0088] With respect to biological activity, an antibody in a formulation retains its biological activity if the antibody in the formulation has the biological activity for its intended use. For example, an antibody in the formulation is considered to have retained its biological activity if the biological activity of the antibody is within about 70% to 130% (e.g., 80% to 130%) of the biological activity exhibited when the formulation was prepared (e.g., as determined by an antigen binding assay).

[0089] As used herein, the term "buffer" refers to a buffered solution that resists changes in pH through the action of its acid-base conjugate components. Examples of such buffers include acetate, succinate, gluconate, histidine, citrate, glycylglycine, and other organic acid buffers.

[0090] The term "histidine buffer" refers to a buffer comprising histidine, wherein the histidine in the buffer can exist alone or in the following form: for example, histidine hydrochloride, histidine acetate, histidine phosphate or histidine sulfate. When histidine is not present alone, a histidine buffer (such as a histidine hydrochloride buffer) can be prepared by titrating L-histidine (free base, solid) with a corresponding acid (such as hydrochloric acid). When histidine exists in a separate form, a pH adjusting agent can be added to obtain a liquid preparation of a suitable pH range according to actual conditions in subsequent preparation steps. In certain preferred embodiments, the histidine in the histidine buffer suitable for the liquid preparation disclosed herein exists alone or in the form of histidine hydrochloride. Beneficial effects

[0091] The present invention provides pharmaceutical compositions of antibodies (e.g., recombinant anti-epidermal growth factor receptor (EGFR) human-mouse chimeric monoclonal antibodies) that meet various needs and have one or more of the following advantages: effectively improving the stability of the antibody under long-term storage conditions without the formation of protein particles; can be directly infused for convenient clinical use; and / or contain high-concentration antibody components for convenient storage and transportation, thereby reducing costs. DETAILED DESCRIPTION

[0092] The present invention will now be described with reference to the following examples which are intended to illustrate the present invention (but not to limit the present invention). Unless otherwise specified, the experiments and methods described in the examples are carried out substantially according to conventional methods well known in the art and described in various references. In the examples, if specific conditions are not specified, they are carried out according to conventional conditions or the conditions recommended by the manufacturer. The reagents or instruments used are conventional products that can be obtained commercially if the manufacturer is not specified. It will be appreciated by those skilled in the art that the examples describe the present invention by way of example and are not intended to limit the scope of protection claimed in the present invention. All published patents and other references mentioned herein are incorporated herein by reference in their entirety.

[0093] 1. Sequence information

[0094] The antibody involved in the Example (hereinafter referred to as "antibody A") has the sequence information described in Table 1 below.

[0095] Table 1: Description of Antibody A Sequences

[0096] 2. Source of reagents

[0097] Antibodies: Antibody A used in the examples is a recombinant human-mouse chimeric monoclonal antibody against epidermal growth factor receptor (EGFR), the amino acid sequence of which is shown in Table 1. Antibody A was expressed in Chinese hamster ovary (CHO) cells and obtained through serum-free cell culture, isolation, and high-level purification.

[0098] Other ingredients: All other ingredients used in the examples are commercially available products.

[0099] 3. Test Method

[0100] The experiments involved in the embodiments respectively adopt the following test methods:

[0101] (1)DSF detection

[0102] The samples were diluted with sample buffer and SYPRO TM Orange Protein Gel Stain was used as a dye, and sample melting curves were collected using an Applied Biosystems 7500 qPCR instrument.

[0103] (2) Polysorbate 80 concentration detection

[0104] Tested according to General Chapter 3203 "Determination of Polysorbate 80 Residues" of the 2020 edition of the "Chinese Pharmacopoeia".

[0105] (3) Insoluble particles

[0106] It is determined in accordance with the general chapter 0903 "Insoluble Particles Examination Method (Light Obstruction Method)" of the 2020 edition of the Chinese Pharmacopoeia.

[0107] (4) Appearance

[0108] Inspection was carried out in a dark room with an illumination of 1000lx-1500lx and direct visual inspection.

[0109] (5) pH value

[0110] Use pH meter to detect.

[0111] (6)SEC-HPLC

[0112] A G3000SWxl column was selected and 1% isopropanol-phosphate buffer was used as the mobile phase for isocratic elution.

[0113] (7)IEC-HPLC

[0114] A WCX-10 chromatographic column was selected, and 10 mM phosphate-100 mM sodium chloride (pH 6.8) was used as the mobile phase to perform gradient elution on the sample to be tested.

[0115] (8) Protein content

[0116] Take 100 μl of sample in a cuvette, measure the absorbance of A279-A320, measure three times and take the average value, and calculate the protein content according to the extinction coefficient.

[0117] Example I-1 Stability of formulations with different buffer systems

[0118] The stability of Formulation I-1-1 and Formulation I-1-2 was tested under high temperature conditions of 40°C ± 2°C / 75% RH ± 5% RH and white light illumination conditions of 4500 lx ± 500 lx.

[0119] (1) The composition of preparation I-1-1 is: Antibody A 2 mg / ml, sodium chloride 8.48 g / L, disodium hydrogen phosphate 1.88 g / L, sodium dihydrogen phosphate 0.41 g / L, pH 7.0;

[0120] (2) The ingredients of formulation I-1-2 are: Antibody A 2 mg / ml, glycine 7.51 g / L, polysorbate 80 0.1 g / L, and citric acid 2.10 g / L. The stability test results are shown in Table 2:

[0121] Table 2 Stability test results of different buffer preparations

[0122] The results showed that the stability of the preparation I-1-1 using a phosphate buffer system was better than that of the preparation I-1-2 using a citrate buffer system under high temperature and light conditions.

[0123] Example 1-2 Stability test of preparations with different surfactant concentrations

[0124] Antibody A preparations I-2-A to I-2-C were prepared according to the following ingredients:

[0125] Preparation I-2-A: Per 1000 ml, contains 2 g of Antibody A, 8.48 g of sodium chloride, 1.88 g of disodium hydrogen phosphate heptahydrate, 0.41 g of sodium dihydrogen phosphate monohydrate, and 0.05 g of polysorbate 80;

[0126] Preparation I-2-B: Per 1000 ml, contains 2 g of Antibody A, 8.48 g of sodium chloride, 1.88 g of disodium hydrogen phosphate heptahydrate, 0.41 g of sodium dihydrogen phosphate monohydrate, and 0.1 g of polysorbate 80;

[0127] Preparation I-2-C: Per 1000 ml, contains 2 g of Antibody A, 8.48 g of sodium chloride, 1.88 g of disodium hydrogen phosphate heptahydrate, 0.41 g of sodium dihydrogen phosphate monohydrate, and 0.2 g of polysorbate 80;

[0128] The insoluble particles of the above preparations were detected. The results are shown in Table 3:

[0129] Table 3 Test results of insoluble particles of the preparations

[0130] In addition, under the conditions of 40° C. and 220 rpm shaking for 3 days, the number of insoluble particles ≥10 μm of the above preparation did not change significantly compared with that on day 0, and met the quality standard for the number of insoluble particles.

[0131] The above results indicate that the preparations of the present invention with different polysorbate 80 concentrations can effectively inhibit the aggregation of antibody A to produce insoluble particles and effectively prevent the formation of protein aggregates.

[0132] Example 1-3 pH screening test

[0133] Antibody A preparations I-3-A to I-3-C were prepared according to the following ingredients:

[0134] I-3-A: Contains Antibody A 2 mg / ml, sodium chloride 8.48 g / L, polysorbate 80 0.1 g, and a sodium dihydrogen phosphate-sodium dihydrogen phosphate buffer system to a pH of 6.1.

[0135] I-3-B: Contains 2 mg / ml of Antibody A, 8.48 g / L of sodium chloride, 0.1 g of polysorbate 80, and a sodium dihydrogen phosphate-sodium dihydrogen phosphate buffer system to a pH of 6.7;

[0136] I-3-C: contains Antibody A 2 mg / ml, sodium chloride 8.48 g / L, polysorbate 80 0.1 g, and a sodium dihydrogen phosphate-sodium dihydrogen phosphate buffer system to a pH of 7.3;

[0137] The DSF method was used to test the exposure temperature (T) of the hydrophobic domain of the antibody preparations at different pH values. h ), the results are shown in Table 4.

[0138] Table 4 T of antibody preparations at different pH h Value Result

[0139] The results showed that in the pH range of 6.1 to 7.3, the exposure temperature of the protein hydrophobic domain (T h ) values ​​showed no significant difference, indicating that the antibodies in the preparations within this pH range had good stability.

[0140] Example I-4. Preparation stability test

[0141] Antibody A Preparation I was prepared according to the following ingredients: per 1000 ml, 2.0 g Antibody A, 8.48 g sodium chloride, 0.41 g sodium dihydrogen phosphate (monohydrate), 1.0 g anhydrous disodium hydrogen phosphate, 0.1 g polysorbate 80, pH 6.7. The stability of Antibody A Preparation I was tested at 40 ± 2°C and 5 ± 3°C under various influencing conditions. The results are shown in Tables 5-7:

[0142] Table 5. Long-term stability data of Antibody A Formulation I at 5±3°C

[0143] In addition, after 48 months of long-term storage at 5±3°C, the biological activity data, number of insoluble particles, and IEC-HPLC test data of Antibody A Preparation I did not show significant differences compared with those on day 0, and all met the quality standards.

[0144] From the above results, it can be seen that the antibody A preparation I of the present invention has good stability under long-term conditions, and there is no significant change in various indicators after storage for 48 months.

[0145] Table 6 Stability data of Antibody A Formulation I under high temperature (40°C ± 2°C) test

[0146] In addition, after 10 days of storage at high temperature (40°C ± 2°C), the biological activity data, number of insoluble particles, and IEC-HPLC test data of Antibody A Preparation I did not show significant differences compared with the data at day 0, and all met the quality standards.

[0147] From the above results, it can be seen that after 10 days of storage at high temperature, all indicators showed no significant changes and met the quality standards, indicating that the antibody A preparation I of the present invention has good stability under high temperature conditions.

[0148] Table 7 Stability data of Antibody A Formulation I under repeated freeze-thaw (-20°C / room temperature) conditions

[0149] As can be seen from the data in Table 7, the antibody A preparation I of the present invention underwent three freeze-thaw cycles at -20°C / room temperature without significant changes in any of the preparation indicators, indicating that the preparation has good stability against repeated freeze-thaw cycles.

[0150] In addition, after storage under light conditions for 10 days, the SEC-HPLC and IEC-HPLC test data of Antibody A Preparation I were not significantly different from those of the light-protected control group, indicating that Antibody A Preparation I of the present invention has good stability under light conditions.

[0151] In addition, after 6 months of storage under accelerated conditions of 25±2°C, the biological activity data, number of insoluble particles, SEC-HPLC, and IEC-HPLC test data of Antibody A Preparation I did not show significant differences compared with those on day 0, and all met the quality standards.

[0152] From the above results, it can be seen that the antibody A preparation I of the present invention has good stability under various influencing factors such as accelerated conditions, long-term conditions, high temperature conditions, light conditions, and repeated freezing and thawing.

[0153] Example I-5. Preparation stability test

[0154] Antibody A formulations I-5-A and I-5-B were formulated according to the following ingredients and their storage stability was tested.

[0155] Antibody A preparation I-5-A: per 1000 ml, Antibody A 2.00 g, sodium chloride 8.48 g, sodium dihydrogen phosphate (monohydrate) 0.41 g, anhydrous disodium hydrogen phosphate 1.00 g, polysorbate 80 0.10 g, pH 6.7.

[0156] Antibody A preparation I-5-B: per 1000 ml, Antibody A 5.00 g, sodium chloride 8.48 g, sodium dihydrogen phosphate (monohydrate) 0.41 g, anhydrous disodium hydrogen phosphate 1.00 g, polysorbate 80 0.10 g, pH 6.7.

[0157] The stability test results are detailed in Table 8:

[0158] Table 8. Stability data of the formulation at 25°C ± 2°C

[0159] As can be seen from the above results, after three months of storage, both the main peak and the aggregate content of the SEC-HPLC chromatograms for both formulations remained unchanged. The low molecular weight content detected by SEC-HPLC chromatography, as well as various parameters detected by IEC-HPLC, non-reduced, and reduced CE-SDS chromatography, showed no significant changes. These results demonstrate that the formulations of the present invention (e.g., formulations I-5-A and I-5-B) exhibit good stability.

[0160] Example I-6. Light stability test

[0161] Antibody A preparation I-6-A and control preparations I and II were formulated according to the following ingredients and tested for their stability under illumination conditions (4500±500 lx, 5±3°C).

[0162] Antibody A preparation I-6-A: per 1000 ml, Antibody A 2.00 g, sodium chloride 8.48 g, sodium dihydrogen phosphate (monohydrate) 0.41 g, anhydrous disodium hydrogen phosphate 1.00 g, polysorbate 80 0.10 g, pH 6.7.

[0163] Control preparation I: per 1000 ml, 5 g of antibody A, 5.84 g of sodium chloride, 7.51 g of glycine, 0.1 g of polysorbate 80, and 2.10 g of citric acid monohydrate, pH 5.3-5.7.

[0164] Control preparation II: per 1000 ml, 2 g of Antibody A, 8.48 g of sodium chloride, 1.88 g of disodium hydrogen phosphate heptahydrate, and 0.41 g of sodium dihydrogen phosphate monohydrate, pH 7.0-7.4.

[0165] Table 9. Light stability test results

[0166] As shown in Table 9, after 7 days of exposure to light, the polymer and monomer contents of the formulations of the present invention (e.g., formulation I-5-A) remained unchanged. However, the polymer content of both control formulations increased significantly, while the monomer content showed a clear downward trend. This indicates that the stability of the formulations of the present invention under light conditions is significantly superior to that of the control formulations.

[0167] Clinical trial examples

[0168] The preparation used in the following clinical trial examples is (hereinafter referred to as "Antibody A Preparation I"): specification 100 mg (50 ml) / bottle, each bottle containing 100 mg of Antibody A, 424 mg of NaCl, 5 mg of polysorbate 80, 20.5 mg of sodium dihydrogen phosphate (monohydrate), 50 mg of anhydrous disodium hydrogen phosphate, and a pH value of 6.7.

[0169] (1) Efficacy and safety of the preparation of the present invention in treating metastatic colorectal cancer

[0170] This study enrolled 18 patients with metastatic colorectal cancer (9 patients in the low-dose group and 9 patients in the high-dose group). The administration route was intravenous injection of Antibody A Preparation I. After the first administration, the patients were observed for 4 weeks and then given the drug once a week. The low-dose group was given a dose of 100 mg / m 2 , 250 mg / m after 4 weeks 2 The high-dose group received the first dose of 400 mg / m 2 , 400 mg / m after 4 weeks 2 or 250 mg / m 2The results of the study showed that the above-mentioned doses had an acceptable safety profile in patients with colorectal cancer. Thirteen subjects underwent efficacy evaluation after treatment, and eight subjects in the high-dose group achieved a partial response (PR), demonstrating that Antibody A Preparation I of the present invention is effective in the treatment of metastatic colorectal cancer.

[0171] (2) Pharmacokinetics and safety of the preparation of the present invention

[0172] In vivo pharmacokinetic and safety studies were conducted on 80 healthy subjects. Antibody A preparation I was administered to 40 subjects in the experimental group at a dose of 250 mg / m 2 The dosage is calculated based on body surface area (BSA). BSA (m 2 )=[(body weight (kg)×height (cm)) / 3600]. Single intravenous administration. The main pharmacokinetic parameter AUC 0- t , and other pharmacokinetic parameters were statistically analyzed, and the results are shown in the following table: Note: AUC 0-t It represents the area under the concentration-time curve from time zero to the last quantifiable concentration.

[0173] The above results show that the pharmacokinetic properties of the preparation of the present invention (e.g., Antibody A Preparation I) are good. In terms of safety, no serious adverse events occurred and no deaths occurred. Most adverse reactions (TEAEs) during treatment occurred a few days after administration, were mild in severity, and most were relieved without any medical intervention. The proportion of subjects with grade 3 or above TEAEs was less than 25%. The subjects' physical and vital signs did not show any changes related to treatment over time, indicating that the preparation of the present invention (e.g., Antibody A Preparation I) has good safety.

[0174] (3) Efficacy of the preparation of the present invention in patients with RAS wild-type colorectal cancer

[0175] A total of 52 patients with RAS wild-type colorectal cancer were enrolled in the study. Patients in the experimental group and the control group received antibody A preparation I+mFOLFOX6 and placebo+mFOLFOX6 as first-line treatment, respectively.

[0176] The median progression-free survival (PFS) of all colorectal cancer patients in the experimental group and the control group was 23.89 months and 14.78 months, respectively. The P value of the PFS rate between all colorectal cancer patients in the experimental group and the control group was 0.0389 (P<0.05), indicating that the difference in PFS between the two groups was statistically significant. The results of the study showed that after treatment of patients with RAS wild-type metastatic colorectal cancer, the experimental group (antibody A preparation I combined with mFOLFOX6) showed an overall better efficacy trend compared with the control group (placebo combined with mFOLFOX6). And the safety results showed that antibody A preparation I has good safety.

[0177] Example II-1 Stability Study of Preparations with Different Ingredients

[0178] Eight groups of antibody A preparations, II-1-1 to II-1-8, were prepared. The pH of each preparation was 5.5. The ingredients of group II-1-1 preparation were: 20 g / L antibody A, 90 g / L sucrose, 0.1 g / L polysorbate 80, and 10 mM citrate buffer. Groups II-1-2 to II-1-8 all used 10 mM histidine salt buffer, which was prepared by mixing histidine and histidine hydrochloride. The ingredients of the preparation in group II-1-3 are: 20 g / L antibody A, 90 g / L sucrose, 0.1 g / L polysorbate 80 and 10 mM histidine buffer; the concentration of antibody A in group II-1-2 is adjusted to 10 g / L based on the ingredients of the preparation in group II-1-3; the concentration of sucrose in groups II-1-4 to II-1-6 is replaced with 8.48 g / L sodium chloride, 28.75 g / L proline, or 50 g / L sorbitol, respectively, based on the ingredients of the preparation in group II-1-3; the concentration of polysorbate 80 in group II-1-7 is adjusted to 0.3 g / L based on the ingredients of the preparation in group II-1-3; the concentration of polysorbate 80 in group II-1-8 is replaced with 0.1 g / L polysorbate 20, based on the ingredients of the preparation in group II-1-3.

[0179] After 28 days of storage at 40°C, the stability of each formulation was tested. The results showed that formulation II-1-1, which used a citric acid buffer, exhibited visible insoluble foreign matter, while formulations II-1-2 to II-1-3, and II-1-6 to II-1-8, which used a histidine buffer, remained colorless, clear liquids. This suggests that formulations with a histidine buffer exhibited improved stability compared to those with a citric acid buffer.

[0180] Furthermore, after 28 days of storage at 40°C, both the II-1-2 preparation (with an Antibody A concentration of 10 g / L) and the II-1-3 preparation (with an Antibody A concentration of 20 g / L) maintained a colorless, clear liquid appearance. Protein content, SEC-HPLC, and IEC-HPLC performance indicators showed no significant differences compared to those at day 0, meeting quality standards. This demonstrates that the preparations at different Antibody A concentrations exhibited excellent stability.

[0181] The remaining stability test results are shown in Tables 10-11:

[0182] Table 10. Stability test results of preparations with different excipients

[0183] As can be seen from the data in Table 10 above, a large amount of smoke-like particles appeared in the appearance of the sodium chloride group (II-1-4) and the proline group (II-1-5) after storage at 40°C for 28 days; the preparations of group II-1-3 and group II-1-6 were more stable, and their appearance remained colorless and clear. In terms of IEC purity, the preparation of group II-1-3 containing sucrose was further superior to the preparation of group II-1-6 containing sorbitol, indicating that the use of sucrose as an excipient is more conducive to the stability of the preparation.

[0184] Table 11. Stability study results of preparations with different polysorbate concentrations and types

[0185] It can be seen from the data in the above table that the II-1-3, II-1-7 and II-1-8 preparations with different concentrations of polysorbate 80 and polysorbate 20 all have good stability.

[0186] Example II-2 Stability test of formulation under long-term and accelerated conditions

[0187] The stability of the II-1-3 group of formulations in Example II-1 was tested under accelerated conditions (25°C, 60% RH±5% RH) and long-term conditions (5°C). The test results are shown in Table 12.

[0188] Table 12. Stability test results of group II-1-3 preparations

[0189] As shown in Table 12, the protein content, SEC-HPLC, and IEC indicators of the injections in Group II-1-3 did not change significantly compared to those at day 0. Furthermore, after 6 months of storage at 5°C, the formulations remained as colorless, clear liquids. These results demonstrate that the formulations of the present invention exhibit good stability under both accelerated and long-term conditions.

[0190] Example II-3 Stability Study of Preparations with Different Antibody Concentrations

[0191] According to the ingredients shown in Table 13 below, three groups of preparations, II-3-1 to II-3-3, were prepared. The stability of each group of preparations under different storage conditions was tested. The test results are shown in Tables 14-15.

[0192] Table 13. Prescription ingredients of each group of Antibody A injection

[0193] Table 14. Stability after 6 months storage at 25°C

[0194] Table 15. Stability after 12 months storage at 5°C

[0195] Stability studies demonstrated that after 12 months at long-term storage (5°C), 30 days at high temperature (40°C), or 6 months at accelerated storage (25°C), the three formulations maintained a colorless, clear liquid appearance. pH, insoluble particulate matter, SEC-HPLC, and IEC-HPLC levels remained similar to those observed at day 0, demonstrating compliance with quality standards. This demonstrates that formulations with antibody concentrations ranging from 20 to 60 mg / ml exhibit excellent stability.

[0196] Example II-4 Stability Study of Preparations at Different pH

[0197] Three formulations, II-4-1 to II-4-3, were prepared. Each formulation contained 60 mg / ml of Antibody A, 80 g / L of sucrose, 0.2 g / L of polysorbate 80, and a 10 mM histidine / histidine hydrochloride buffer. The pH values ​​for the three formulations were 5.5 for II-4-1, 5.2 for II-4-2, and 5.8 for II-4-3. The stability of each formulation under different storage conditions was tested. The results are shown in Table 16.

[0198] Table 16. High temperature stability test results of preparations with different pH values

[0199] In addition, the IEC-HPLC purity of the three groups of preparations was not significantly different from that at day 0, and met the quality standards. The above experimental results show that the preparations in each group with pH 5.2-5.8 have good stability.

[0200] Example II-5: Stability of preparations under different conditions

[0201] Antibody A Formulation II was prepared according to the following ingredients: Antibody A 20 g / L, histidine buffer 20 mmol / L, polysorbate 80 0.2 g / L, sucrose 80 g / L, pH 5.5 ± 0.3. The stability of the formulation was tested under various conditions:

[0202] Table 17. Stability of Antibody A Formulation II

[0203] Note: AG4h: 4 hours of shaking treatment at room temperature; -70℃ FT4X: 4 cycles of freeze-thaw at -70℃ / room temperature; 5℃ 1M: 1 month at 5℃.

[0204] In addition, after antibody A preparation II was placed at 40°C for 28 days, or irradiated with white light 4500lx±500lx for 2 weeks, all test indicators remained stable compared with those on day 0, with no significant differences.

[0205] Based on the above data, it can be seen that the various test indicators of Antibody A Preparation II of the present invention did not show significant differences compared with those at day 0 under various conditions such as high temperature, oscillation, and repeated freeze-thaw cycles, and all met the quality standards, indicating that the preparation of the present invention exhibits good stability against various influencing factors.

[0206] The above-described embodiments merely illustrate several implementations of the present invention, and while their descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the patent. It should be noted that a person skilled in the art would be able to make numerous variations and improvements without departing from the spirit of the present invention, all of which fall within the scope of protection of the present invention. Therefore, the scope of protection of the patent for this invention shall be determined by the appended claims.

Claims

1. A pharmaceutical composition, characterized in that It contains 0.5-10 g / L of antibody, 6-12 g / L of stabilizer, 0.01-0.5 g / L of surfactant, and a buffer system; the pH of the liquid preparation is 6.0-8.0; The stabilizer is selected from one or more of sodium chloride, glycine, sucrose, sorbitol, and proline; The surfactant is selected from polysorbate 20 or polysorbate 80; The buffer system is selected from a phosphate buffer system, a citrate buffer system or a histidine buffer system.

2. The pharmaceutical composition according to claim 1, comprising 1 to 8 g / L of the antibody, preferably 1 to 3 g / L or 4 to 6 g / L of the antibody.

3. The pharmaceutical composition according to claim 1 or 2, comprising 0.5-5 g / L of antibody, 6-12 g / L of stabilizer, 0.01-0.5 g / L of surfactant, and a buffer system; the pH of the pharmaceutical composition is 6.0-8.

0.

4. The pharmaceutical composition according to any one of claims 1-3, comprising 1.8-2.2 g / L of antibody, 7.632-9.328 g / L of stabilizer, 0.05-0.15 g / L of surfactant, and a buffer system; the pH of the pharmaceutical composition is 6.4-7.0; or, the pharmaceutical composition comprises 4-6 g / L of antibody, 7-10 g / L of stabilizer, 0.05-0.15 g / L of surfactant, and a buffer system; the pH of the pharmaceutical composition is 6.4-7.

0.

5. The pharmaceutical composition according to any one of claims 1 to 4, comprising 1.8 to 2.2 g / L of antibody, 7.632 to 9.328 g / L of NaCl, 0.05 to 0.15 g / L of polysorbate 80, 0.369 to 0.451 g / L of sodium dihydrogen phosphate, and 0.9 to 1.1 g / L of sodium dihydrogen phosphate; the pH of the pharmaceutical composition is 6.4 to 7.0; or, the pharmaceutical composition comprises 4 to 6 g / L of antibody, 7.632 to 9.328 g / L of NaCl, 0.05 to 0.15 g / L of polysorbate 80, 0.369 to 0.451 g / L of sodium dihydrogen phosphate, and 0.9 to 1.1 g / L of sodium dihydrogen phosphate; the pH of the pharmaceutical composition is 6.4 to 7.

0.

6. The pharmaceutical composition according to any one of claims 1 to 5, comprising 2.0 g of anti-EGFR antibody, 8.48 g of sodium chloride, 0.41 g of sodium dihydrogen phosphate (monohydrate), 1.0 g of sodium dihydrogen phosphate, and 0.1 g of polysorbate 80; the pH of the pharmaceutical composition is 6.7; or, the pharmaceutical composition comprises 5.0 g of anti-EGFR antibody, 8.48 g of sodium chloride, 0.41 g of sodium dihydrogen phosphate (monohydrate), 1.0 g of anhydrous sodium dihydrogen phosphate, and 0.1 g of polysorbate 80; the pH of the liquid preparation is 6.

7.

7. A pharmaceutical composition, characterized in that It contains 10-100 g / L of antibody, 50-100 g / L of stabilizer, 0.05-1.0 g / L of surfactant, and a buffer system; the pH of the pharmaceutical composition is 5.0-7.5; The stabilizer is selected from one or more of sodium chloride, glycine, sucrose, sorbitol, and proline; The surfactant is selected from polysorbate 20 or polysorbate 80; The buffer system is selected from a phosphate buffer system, a citrate buffer system or a histidine buffer system.

8. The pharmaceutical composition according to claim 7, comprising 20-60 g / L of antibody, 0.1-0.3 g / L of surfactant, and 80-90 g / L of stabilizer, and the pH of the pharmaceutical composition is 5.2-6.

2.

9. The pharmaceutical composition according to claim 7 or 8, comprising 20-60 g / L of antibody, 10-20 mmol / L of histidine and histidine hydrochloride in total, 0.1-0.3 g / L of polysorbate 80, and 80-90 g / L of sucrose per 1000 ml, and the pH of the pharmaceutical composition is 5.2-6.

2.

10. The pharmaceutical composition of claim 9, comprising 20 g / L anti-EGFR antibody, 0.67 g / L histidine, 3.29 g / L histidine hydrochloride, 0.2 g / L polysorbate 80, and 80 g / L sucrose per 1000 ml, and the pH of the pharmaceutical composition is 5.5±0.

3.

11. The pharmaceutical composition according to any one of claims 1 to 10, wherein The antibody is an anti-EGFR antibody.

12. The pharmaceutical composition according to claim 11, wherein The anti-EGFR antibody comprises heavy chain CDRs as shown in SEQ ID NOs: 1-3, respectively, and light chain CDRs as shown in SEQ ID NOs: 4-6, respectively.

13. The pharmaceutical composition according to claim 12, wherein The anti-EGFR antibody has a heavy chain variable region as shown in SEQ ID NO:7 and a light chain variable region as shown in SEQ ID NO:

8.

14. The pharmaceutical composition according to claim 13, wherein The anti-EGFR antibody has a heavy chain as shown in SEQ ID NO:9 and a light chain as shown in SEQ ID NO:

10.

15. A solid preparation prepared by freeze-drying the pharmaceutical composition according to any one of claims 1 to 14.

16. A product comprising a container filled with the pharmaceutical composition according to any one of claims 1 to 14 or the solid preparation according to claim 15.

17. The article of claim 16, wherein: The container is a glass bottle, a metal alloy container or a prefilled syringe.

18. A pharmaceutical preparation, which is obtained by diluting the pharmaceutical composition according to any one of claims 7 to 14 with a buffer.

19. The pharmaceutical preparation according to claim 18, which is obtained by diluting the pharmaceutical composition according to any one of claims 7 to 14 with histidine hydrochloride-histidine buffer.

20. The pharmaceutical preparation according to claim 19, wherein the liquid preparation is obtained by diluting the pharmaceutical composition according to any one of claims 7 to 14 by 5 to 50 times with histidine hydrochloride-histidine buffer.

21. The pharmaceutical preparation according to claim 20, wherein the liquid preparation is obtained by diluting the pharmaceutical composition according to any one of claims 7 to 14 10 times with histidine hydrochloride-histidine buffer.

22. The pharmaceutical preparation according to any one of claims 19 to 21, wherein the pH of the histidine hydrochloride-histidine buffer is 5.2-6.

2.

23. A method for stabilizing an antibody, characterized in that: The method is carried out by combining the antibody, stabilizer, surfactant and / or buffer system described in any one of claims 1 to 14, and controlling the pH of the combination within the range described in any one of claims 1 to 10.

24. Use of the pharmaceutical composition according to any one of claims 1 to 14, the solid preparation according to claim 15, the product according to claim 16 or 17, or the pharmaceutical preparation according to any one of claims 18 to 22 in the preparation of a drug, wherein the drug is used for: (a) prevention and / or treatment of tumors; (b) adjuvant therapy for tumors; (c) diagnosis of tumors; (d) Any combination of (a)-(c).

25. The use according to claim 24, characterized in that The drug is administered separately, in combination, simultaneously, or sequentially with another pharmaceutically active agent; optionally, the other pharmaceutically active agent is a chemotherapeutic drug.

26. The use according to claim 24 or 25, characterized in that The tumor is selected from breast cancer, lung cancer, such as non-small cell lung cancer or squamous cell lung cancer, liver cancer, stomach cancer, intestinal cancer, such as colon cancer or rectal cancer, esophageal cancer, ovarian cancer, cervical cancer, kidney cancer, prostate cancer, bladder cancer, pancreatic cancer, Merkel cell carcinoma, bile duct cancer, nasopharyngeal carcinoma, head and neck squamous cell carcinoma, glioma, melanoma, leukemia and lymphoma.

27. Use of the pharmaceutical composition of any one of claims 1 to 14, the solid preparation of claim 15 or the pharmaceutical preparation of any one of claims 18 to 22 in the preparation of a liquid preparation for injection.

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