Pharmaceutical composition comprising Anti-PD-l1 antibody and pharmaceutical use thereof

Abstract

The present disclosure relates to a pharmaceutical composition comprising an anti-PD-L1 antibody and pharmaceutical use thereof. Specifically, the present disclosure relates to a pharmaceutical composition comprising an anti-PD-L1 antibody and a hyaluronidase, and use thereof in the treatment of tumors.

Description

A pharmaceutical composition comprising an anti-PD-L1 antibody and its pharmaceutical uses.

[0001] This application claims priority to Chinese patent application CN202510118513.0 filed on January 24, 2025 and Chinese patent application CN202511919411.5 filed on December 18, 2025. Technical Field

[0002] This disclosure pertains to the field of pharmaceutical formulations, specifically relating to a pharmaceutical composition comprising an anti-PD-L1 antibody and its pharmaceutical uses. Background Technology

[0003] The statements herein are provided only as background information in connection with this disclosure and do not necessarily constitute prior art.

[0004] Compared to other chemical drugs, antibody drugs have larger molecular weights and more complex structures, making them more susceptible to degradation, polymerization, or unwanted chemical modifications, thus becoming unstable. During manufacturing, storage, and delivery, antibody drugs undergo physical and chemical degradation. These instabilities can reduce the potency of antibody drugs and increase the risk of adverse events in patients, thereby affecting regulatory approval (see, for example, Wang et al., J. Pharm. Sci. 96: 1, 2007).

[0005] In many cases, antibodies are administered via intravenous (IV) injection or infusion. Unfortunately, the amount of antibody that can be administered intravenously is limited by the antibody's physicochemical properties, particularly its solubility and stability in suitable liquid formulations and the volume of the infusion solution. Alternative routes of administration are subcutaneous or intramuscular injection.

[0006] Subcutaneous injection offers advantages such as reduced drug preparation, infusion, and observation time, thus saving time for patients and healthcare workers, reducing the occupation of medical resources, and lowering medical costs. Some subcutaneous formulations can achieve a fixed dosage, eliminating the need for dosage calculation and preparation, significantly reducing the risk of dosage errors. Furthermore, some subcutaneous formulations have been reported to reduce the risk of infusion-related reactions and infections. They also save time and improve patient compliance. However, subcutaneous injection is limited by bioavailability and injection volume. To meet the requirements of subcutaneous injection, a high-concentration antibody formulation is needed. Therefore, the development of stable, high-concentration subcutaneous antibody formulations is essential for the success of antibody drugs.

[0007] However, formulating high-concentration antibody preparations presents numerous challenges. For example, increasing antibody concentration often involves challenges related to protein aggregation, solubility, stability, and viscosity (see, for example, Shire et al., J. Pharm. Sci. 93: 1390, 2004). The proteins in these drugs often exhibit microscopic heterogeneity, or "heterogeneity," including isomers related to charge, hydrophobicity, and morphology. Heterogeneity caused by differences in the charge carried by protein molecules is called charge isomerism, which is mainly related to post-translational modifications and may originate from complex biosynthetic pathways of protein molecules, such as cell lines and culture processes.

[0008] The charge heterogeneity of proteins can significantly impact the stability, efficacy, immunogenicity, and pharmacokinetics of protein drugs, making it a key quality attribute (CQA) for protein drugs. For example, Genentech summarized the effects of charge changes on the efficacy and pharmacokinetics of marketed and preclinical antibody drugs. The results showed that: ① Charge variation exceeding one pH unit affects drug tissue distribution and pharmacokinetics; ② Increased positive charge increases tissue retention and decreases blood clearance; ③ Decreased positive charge reduces tissue retention and increases systemic clearance. Furthermore, protein charge heterogeneity can also reflect the stability of the protein drug manufacturing process. In conclusion, detecting protein charge heterogeneity is crucial for the quality control of protein drugs. This necessitates continuous innovative process development to ensure that high-concentration formulations can stably and efficiently exert their therapeutic effects.

[0009] Programmed death molecule-1 (PD-1), discovered in 1992, is a protein receptor expressed on the surface of T cells and participates in the apoptosis process. PD-1 has two ligands, PD-L1 and PD-L2. PD-L1 inhibits the immune system by binding to PD-1 and B7-1, and many tumor cells and immune cells in the tumor tissue microenvironment express PD-L1. New research has found high expression of PD-L1 protein in human tumor tissues such as breast cancer, lung cancer, gastric cancer, colorectal cancer, kidney cancer, melanoma, non-small cell lung cancer, colon cancer, bladder cancer, ovarian cancer, pancreatic cancer, and liver cancer, and the expression level of PD-L1 is closely related to the patient's clinical condition and prognosis. Because PD-L1 acts as a second signaling pathway to inhibit T cell proliferation, blocking the binding between PD-L1 and PD-1 has become a very important target in the field of tumor immunotherapy.

[0010] Formulations of anti-PD-L1 antibodies that can be administered intravenously have been developed (see WO2018210230A1). (Adebelimab) was launched in China in 2023, containing an anti-PD-L1 antibody, sucrose, succinic acid, and polysorbate 80. However, the optimal formulation of the anti-PD-L1 antibody (adebelimab) suitable for subcutaneous injection has not yet been developed, and significant unmet medical needs remain. Summary of the Invention

[0011] This disclosure provides a pharmaceutical composition containing an anti-PD-L1 antibody that is more conducive to production and administration and has stable performance. The pharmaceutical composition has advantages such as good stability, high bioavailability, inhibition of components with charge heterogeneity, and inhibition of aggregates.

[0012] In some embodiments, this disclosure provides a pharmaceutical composition as shown in any of the following:

[0013] (1) A pharmaceutical composition comprising an anti-PD-L1 antibody, a buffer solution, a sugar, a surfactant, and an antioxidant;

[0014] (2) A pharmaceutical composition comprising an anti-PD-L1 antibody, a buffer solution, a sugar, a surfactant, an antioxidant, and hyaluronidase; or

[0015] (3) A pharmaceutical composition comprising an anti-PD-L1 antibody and hyaluronidase; or

[0016] (4) A pharmaceutical composition comprising an anti-PD-L1 antibody, a buffer solution and hyaluronidase.

[0017] Anti-PD-L1 antibody

[0018] In some embodiments, the anti-PD-L1 antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3; wherein the heavy chain variable region HCDR1 comprises the amino acid sequence of SEQ ID NO: 1, HCDR2 comprises the amino acid sequence of SEQ ID NO: 2, and HCDR3 comprises the amino acid sequence of SEQ ID NO: 3, and the light chain variable region LCDR1 comprises the amino acid sequence of SEQ ID NO: 4, LCDR2 comprises the amino acid sequence of SEQ ID NO: 5, and LCDR3 comprises the amino acid sequence of SEQ ID NO: 6.

[0019] In some implementations, the anti-PD-L1 antibody is a murine antibody, a chimeric antibody, a humanized antibody, or a fully human antibody.

[0020] In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the anti-PD-L1 antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 7, or an amino acid sequence having at least 80% (e.g., at least 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with it; and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 8, or an amino acid sequence having at least 80% (e.g., at least 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with it. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the anti-PD-L1 antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 7, and the light chain variable region comprising the amino acid sequence of SEQ ID NO: 8.

[0021] In some embodiments, the anti-PD-L1 antibody further comprises a heavy chain constant region and / or a light chain constant region. In some embodiments, the heavy chain constant region of the anti-PD-L1 antibody is a constant region derived from human IgG1, IgG2, IgG3, or IgG4. In some embodiments, the anti-PD-L1 antibody is a heavy chain constant region derived from human IgG2 or IgG4. In some embodiments, the anti-PD-L1 antibody comprises an IgG4 heavy chain constant region incorporating F234A and L235A mutations. In some embodiments, the light chain constant region is a light chain constant region derived from human κ or λ chains.

[0022] In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the anti-PD-L1 antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 9, or an amino acid sequence having at least 80% (e.g., at least 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity therewith; and the light chain comprises the amino acid sequence of SEQ ID NO: 10, or an amino acid sequence having at least 80% (e.g., at least 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity therewith. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the anti-PD-L1 antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 9, and the light chain comprises the amino acid sequence of SEQ ID NO: 10. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the anti-PD-L1 antibody is adebelimab (HRP00052).

[0023] In some embodiments, such as the pharmaceutical composition described in any of the preceding embodiments, the anti-PD-L1 antibody may be prepared by the method described in the example of WO2017084495A1.

[0024] In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of the anti-PD-L1 antibody is 80 mg / mL or higher. In some embodiments, the concentration of the anti-PD-L1 antibody is 100 mg / mL or higher. In some embodiments, the concentration of the anti-PD-L1 antibody is from 80 mg / mL to 200 mg / mL. In some embodiments, the concentration of the anti-PD-L1 antibody is from 80 mg / mL to 180 mg / mL. In some embodiments, the concentration of the anti-PD-L1 antibody is from 100 mg / mL to 180 mg / mL. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of the anti-PD-L1 antibody is from 120 mg / mL to 180 mg / mL. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of the anti-PD-L1 antibody is from 135 mg / mL to 165 mg / mL. In some embodiments, the concentration of the anti-PD-L1 antibody is about 80 mg / mL, about 85 mg / mL, about 90 mg / mL, about 95 mg / mL, about 100 mg / mL, about 105 mg / mL, about 110 mg / mL, about 115 mg / mL, about 120 mg / mL, about 125 mg / mL, about 130 mg / mL, about 135 mg / mL, about 140 mg / mL, about 145 mg / mL, about 150 mg / mL, about 155 mg / mL, about 160 mg / mL, about 165 mg / mL, about 170 mg / mL, about 175 mg / mL, about 180 mg / mL, about 185 mg / mL, about 190 mg / mL, about 195 mg / mL, or about 200 mg / mL. In some embodiments, in the pharmaceutical composition as described in any of the preceding claims, the concentration of the anti-PD-L1 antibody is about 150 mg / mL. In some embodiments, the concentration of the anti-PD-L1 antibody is 80 mg / mL, 85 mg / mL, 90 mg / mL, 95 mg / mL, 100 mg / mL, 105 mg / mL, 110 mg / mL, 115 mg / mL, 120 mg / mL, 125 mg / mL, 130 mg / mL, 135 mg / mL, 140 mg / mL, 145 mg / mL, 150 mg / mL, 155 mg / mL, 160 mg / mL, 165 mg / mL, 170 mg / mL, 175 mg / mL, 180 mg / mL, 185 mg / mL, 190 mg / mL, 195 mg / mL, or 200 mg / mL, or any range between these values. In some embodiments, in the pharmaceutical composition as described in any of the preceding claims, the concentration of the anti-PD-L1 antibody is 150 mg / mL.

[0025] Buffer system

[0026] In some embodiments, the pharmaceutical composition as described in any of the preceding claims further comprises a buffer solution. In some embodiments, the pharmaceutical composition as described in any of the preceding claims contains the buffer solution as a histidine-histidine hydrochloride buffer, a histidine-histidine acetate buffer, or a succinate-sodium succinate buffer. In some embodiments, the pharmaceutical composition as described in any of the preceding claims contains the buffer solution as a histidine-histidine hydrochloride buffer.

[0027] In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of the buffer solution is from 1 mM to 100 mM. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of the buffer solution is from 5 mM to 50 mM. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of the buffer solution is from 5 mM to 40 mM. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of the buffer solution is from 10 mM to 40 mM. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of the buffer solution is from 10 mM to 30 mM. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of the buffer solution is from 15 mM to 30 mM. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of the buffer solution is from 15 mM to 25 mM. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of the buffer solution is about 20 mM. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of the buffer solution is about 30 mM. In some embodiments, the concentration of the buffer solution is about 1 mM, about 5 mM, about 10 mM, about 15 mM, about 20 mM, about 25 mM, about 30 mM, about 35 mM, about 36 mM, about 37 mM, about 38 mM, about 39 mM, about 40 mM, about 40.5 mM, about 41 mM, about 42 mM, about 43 mM, about 44 mM, about 45 mM, about 46 mM, about 47 mM, about 48 mM, about 49 mM, about 49.5 mM, about 50 mM, about 55 mM, about 60 mM, about 65 mM, about 70 mM, about 75 mM, about 80 mM, about 85 mM, about 90 mM, about 95 mM, or about 100 mM. In some embodiments, the concentration of the buffer solution is 1 mM, 5 mM, 10 mM, 15 mM, 20 mM, 25 mM, 30 mM, 35 mM, 36 mM, 37 mM, 38 mM, 39 mM, 40 mM, 40.5 mM, 41 mM, 42 mM, 43 mM, 44 mM, 45 mM, 46 mM, 47 mM, 48 mM, 49 mM, 49.5 mM, 50 mM, 55 mM, 60 mM, 65 mM, 70 mM, 75 mM, 80 mM, 85 mM, 90 mM, 95 mM, or 100 mM, or any range between these values. In some embodiments, the concentration of the buffer solution is 20 mM. In some embodiments, the concentration of the buffer solution is 30 mM.

[0028] In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of the histidine-histidine hydrochloride buffer is from 1 mM to 100 mM. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of the histidine-histidine hydrochloride buffer is from 5 mM to 50 mM. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of the histidine-histidine hydrochloride buffer is from 5 mM to 40 mM. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of the histidine-histidine hydrochloride buffer is from 10 mM to 40 mM. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of the histidine-histidine hydrochloride buffer is from 10 mM to 30 mM. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of the histidine-histidine hydrochloride buffer is from 15 mM to 30 mM. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of the histidine-histidine hydrochloride buffer is from 15 mM to 25 mM. In some embodiments, the concentration of the histidine-histidine hydrochloride buffer solution is about 20 mM in the pharmaceutical composition as described in any of the preceding embodiments. In some embodiments, the concentration of the histidine-histidine hydrochloride buffer solution is about 30 mM in the pharmaceutical composition as described in any of the preceding embodiments. In some embodiments, the concentration of the histidine-histidine hydrochloride buffer is about 1 mM, about 5 mM, about 10 mM, about 15 mM, about 20 mM, about 25 mM, about 30 mM, about 35 mM, about 36 mM, about 37 mM, about 38 mM, about 39 mM, about 40 mM, about 40.5 mM, about 41 mM, about 42 mM, about 43 mM, about 44 mM, about 45 mM, about 46 mM, about 47 mM, about 48 mM, about 49 mM, about 49.5 mM, about 50 mM, about 55 mM, about 60 mM, about 65 mM, about 70 mM, about 75 mM, about 80 mM, about 85 mM, about 90 mM, about 95 mM, or about 100 mM. In some embodiments, the concentration of the histidine-histidine hydrochloride buffer is 1 mM, 5 mM, 10 mM, 15 mM, 20 mM, 25 mM, 30 mM, 35 mM, 36 mM, 37 mM, 38 mM, 39 mM, 40 mM, 40.5 mM, 41 mM, 42 mM, 43 mM, 44 mM, 45 mM, 46 mM, 47 mM, 48 mM, 49 mM, 49.5 mM, 50 mM, 55 mM, 60 mM, 65 mM, 70 mM, 75 mM, 80 mM, 85 mM, 90 mM, 95 mM, or 100 mM, or any range between these values. In some embodiments, the concentration of the histidine-histidine hydrochloride buffer is 20 mM. In some embodiments, the concentration of the histidine-histidine hydrochloride buffer is 30 mM.

[0029] In some embodiments, the pH of the pharmaceutical composition is 5.0 to 6.5. In some embodiments, the pH of the pharmaceutical composition is 5.0 to 5.8. In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments has a pH of 5.0 to 5.6. In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments has a pH of 5.0 to 5.5. In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments has a pH of 5.2 to 5.4. In some embodiments, the pH of the pharmaceutical composition is about 5.0, about 5.1, about 5.2, about 5.3, about 5.4, about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, about 6.0, about 6.1, about 6.2, about 6.3, about 6.4, or about 6.5. In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments has a pH of about 5.2. In some embodiments, the pharmaceutical composition as described in any of the preceding claims has a pH of about 5.3. In some embodiments, the pharmaceutical composition has a pH of 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, or 6.5, or any range between these values. In some embodiments, the pharmaceutical composition as described in any of the preceding claims has a pH of 5.2. In some embodiments, the pharmaceutical composition as described in any of the preceding claims has a pH of 5.3. When point values ​​are mentioned in this disclosure, it should be understood that the point value includes a range of error. This range of error is due to factors such as laboratory environment, human operation, instrumentation, methodology, measurement error, etc. For example, when the pH value is about 5.3, it should be understood that it includes a range of error.

[0030] sugar

[0031] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments includes a sugar. In some embodiments, the sugar is selected from conventional compositions (CH2O). n And its derivatives, including monosaccharides, disaccharides, trisaccharides, polysaccharides, sugar alcohols, reducing sugars, non-reducing sugars, etc. In some embodiments, the sugar is selected from sucrose, trehalose, glucose, lactose, fructose, maltose, dextran, glycerol, erythritol, glycerol, arabinitol, sylitol, sorbitol, mannitol, melitriose, maltotriose, stachyose, maltose, lactulose, maltitol, maltitol, lactitol, and isomaltulose, etc. In some embodiments, the sugar is sucrose, trehalose, mannitol, or sorbitol. In some embodiments, the sugar is sucrose.

[0032] In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the sugar concentration is from 10 mg / mL to 120 mg / mL. In some embodiments, the sugar concentration is from 30 mg / mL to 90 mg / mL. In some embodiments, the sugar concentration is from 50 mg / mL to 90 mg / mL. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the sugar concentration is from 55 mg / mL to 85 mg / mL. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the sugar concentration is from 65 mg / mL to 75 mg / mL. In some embodiments, the sugar concentration is from 55 mg / mL to 65 mg / mL. In some embodiments, the sugar concentration is about 10 mg / mL, about 20 mg / mL, about 30 mg / mL, about 40 mg / mL, about 45 mg / mL, about 50 mg / mL, about 55 mg / mL, about 60 mg / mL, about 65 mg / mL, about 66 mg / mL, about 67 mg / mL, about 68 mg / mL, about 69 mg / mL, about 70 mg / mL, about 71 mg / mL, about 72 mg / mL, about 73 mg / mL, about 74 mg / mL, about 75 mg / mL, about 80 mg / mL, about 85 mg / mL, about 90 mg / mL, about 95 mg / mL, about 100 mg / mL, about 110 mg / mL, or about 120 mg / mL. In some embodiments, the sugar concentration is about 70 mg / mL. In some embodiments, the sugar concentration is about 75 mg / mL. In some embodiments, the sugar concentration is 10 mg / mL, 20 mg / mL, 30 mg / mL, 40 mg / mL, 45 mg / mL, 50 mg / mL, 55 mg / mL, 60 mg / mL, 65 mg / mL, 66 mg / mL, 67 mg / mL, 68 mg / mL, 69 mg / mL, 70 mg / mL, 71 mg / mL, 72 mg / mL, 73 mg / mL, 74 mg / mL, 75 mg / mL, 80 mg / mL, 85 mg / mL, 90 mg / mL, 95 mg / mL, 100 mg / mL, 110 mg / mL, or 120 mg / mL, or any range between these values. In some embodiments, the sugar concentration is 70 mg / mL. In some embodiments, the sugar concentration is 75 mg / mL.

[0033] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments, wherein the concentration of sucrose is from 10 mg / mL to 120 mg / mL. In some embodiments, the concentration of sucrose is from 30 mg / mL to 90 mg / mL. In some embodiments, the concentration of sucrose is from 50 mg / mL to 90 mg / mL. In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments, wherein the concentration of sucrose is from 55 mg / mL to 85 mg / mL. In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments, wherein the concentration of sucrose is from 65 mg / mL to 75 mg / mL. In some embodiments, the concentration of sucrose is from 55 mg / mL to 65 mg / mL. In some embodiments, the concentration of the sucrose is about 10 mg / mL, about 20 mg / mL, about 30 mg / mL, about 40 mg / mL, about 45 mg / mL, about 50 mg / mL, about 55 mg / mL, about 60 mg / mL, about 65 mg / mL, about 66 mg / mL, about 67 mg / mL, about 68 mg / mL, about 69 mg / mL, about 70 mg / mL, about 71 mg / mL, about 72 mg / mL, about 73 mg / mL, about 74 mg / mL, about 75 mg / mL, about 80 mg / mL, about 85 mg / mL, about 90 mg / mL, about 95 mg / mL, about 100 mg / mL, about 110 mg / mL, or about 120 mg / mL. In some embodiments, the concentration of the sucrose is about 70 mg / mL. In some embodiments, the concentration of the sucrose is about 75 mg / mL. In some embodiments, the concentration of sucrose is 10 mg / mL, 20 mg / mL, 30 mg / mL, 40 mg / mL, 45 mg / mL, 50 mg / mL, 55 mg / mL, 60 mg / mL, 65 mg / mL, 66 mg / mL, 67 mg / mL, 68 mg / mL, 69 mg / mL, 70 mg / mL, 71 mg / mL, 72 mg / mL, 73 mg / mL, 74 mg / mL, 75 mg / mL, 80 mg / mL, 85 mg / mL, 90 mg / mL, 95 mg / mL, 100 mg / mL, 110 mg / mL, or 120 mg / mL, or any range between these values. In some embodiments, the concentration of sucrose is 70 mg / mL. In some embodiments, the concentration of sucrose is 75 mg / mL. In some embodiments, when the sucrose concentration is 70 mg / mL, the osmotic pressure is isotonic median. In some implementations, when the sucrose concentration is 70 mg / mL, the osmotic pressure of the formulation is measured to be 306 mOsm / kg.

[0034] surfactants

[0035] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments further comprises a surfactant. In some embodiments, the surfactant is a nonionic surfactant. In some embodiments, the surfactant is selected from polysorbates (e.g., polysorbate 80, polysorbate 20), poloxamer (e.g., poloxamer 188, i.e., PF68 or P188), Triton, sodium lauryl sulfonate, sodium lauryl sulfonate, sodium octyl glycoside, lauryl-sulfobetaine, myristyl-sulfobetaine, linoleyl-sulfobetaine, stearyl-sulfobetaine, lauryl-sarcosine, myristyl-sarcosine, linoleyl-sarcosine, stearyl-sarcosine, linoleyl-saccharide, succinate ... Betaine, myristyl-betaine, cetyl-betaine, lauramidopropyl-betaine, cocarbamate-propyl-betaine, linoleamide-propyl-betaine, myristamidopropyl-betaine, palmitoamide-propyl-betaine, isostearamidopropyl-betaine, myristamidopropyl-dimethylamine, palmitoamide-propyl-dimethylamine, isostearamidopropyl-dimethylamine, sodium methyl cocoyl, sodium methyl oleate, polyethylene glycol, polypropylene glycol, copolymers of ethylene and propylene glycol, etc. In some embodiments, the surfactant is polysorbate. In some embodiments, the surfactant is polysorbate 80 (PS80) or polysorbate 20 (PS20). In some embodiments, the surfactant is polysorbate 80 (PS80).

[0036] In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of the surfactant is from 0.01 mg / mL to 2.0 mg / mL. In some embodiments, the concentration of the surfactant is from 0.1 mg / mL to 1.0 mg / mL. In some embodiments, the concentration of the surfactant is from 0.1 mg / mL to 0.8 mg / mL. In some embodiments, the concentration of the surfactant is from 0.1 mg / mL to 0.6 mg / mL. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of the surfactant is from 0.4 mg / mL to 0.8 mg / mL. In some embodiments, the surfactant concentration is about 0.01 mg / mL, about 0.02 mg / mL, about 0.05 mg / mL, about 0.1 mg / mL, about 0.2 mg / mL, about 0.3 mg / mL, about 0.4 mg / mL, about 0.5 mg / mL, about 0.6 mg / mL, about 0.7 mg / mL, about 0.8 mg / mL, about 0.9 mg / mL, about 1.0 mg / mL, about 1.5 mg / mL, or about 2.0 mg / mL. In some embodiments, the surfactant concentration is about 0.1 mg / mL. In some embodiments, the surfactant concentration is about 0.6 mg / mL. In some embodiments, the surfactant concentration is 0.01 mg / mL, 0.02 mg / mL, 0.05 mg / mL, 0.1 mg / mL, 0.2 mg / mL, 0.3 mg / mL, 0.4 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, 1.5 mg / mL, or 2.0 mg / mL, or any range between these values. In some embodiments, the surfactant concentration is 0.1 mg / mL. In some embodiments, the surfactant concentration is 0.6 mg / mL.

[0037] In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of polysorbate 80 is from 0.01 mg / mL to 2.0 mg / mL. In some embodiments, the concentration of polysorbate 80 is from 0.1 mg / mL to 1.0 mg / mL. In some embodiments, the concentration of polysorbate 80 is from 0.1 mg / mL to 0.8 mg / mL. In some embodiments, the concentration of polysorbate 80 is from 0.1 mg / mL to 0.6 mg / mL. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of polysorbate 80 is from 0.4 mg / mL to 0.8 mg / mL. In some embodiments, the concentration of polysorbate 80 is about 0.01 mg / mL, about 0.02 mg / mL, about 0.05 mg / mL, about 0.1 mg / mL, about 0.2 mg / mL, about 0.3 mg / mL, about 0.4 mg / mL, about 0.5 mg / mL, about 0.6 mg / mL, about 0.7 mg / mL, about 0.8 mg / mL, about 0.9 mg / mL, about 1.0 mg / mL, about 1.5 mg / mL, or about 2.0 mg / mL. In some embodiments, the concentration of polysorbate 80 is about 0.1 mg / mL. In some embodiments, the concentration of polysorbate 80 is about 0.6 mg / mL. In some embodiments, the concentration of polysorbate 80 is 0.01 mg / mL, 0.02 mg / mL, 0.05 mg / mL, 0.1 mg / mL, 0.2 mg / mL, 0.3 mg / mL, 0.4 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, 1.5 mg / mL, or 2.0 mg / mL, or any range between these values. In some embodiments, the concentration of polysorbate 80 is 0.1 mg / mL. In some embodiments, the concentration of polysorbate 80 is 0.6 mg / mL.

[0038] In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of polysorbate 20 is from 0.01 mg / mL to 2.0 mg / mL. In some embodiments, the concentration of polysorbate 20 is from 0.1 mg / mL to 1.0 mg / mL. In some embodiments, the concentration of polysorbate 20 is from 0.1 mg / mL to 0.8 mg / mL. In some embodiments, the concentration of polysorbate 20 is from 0.1 mg / mL to 0.6 mg / mL. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of polysorbate 20 is from 0.4 mg / mL to 0.8 mg / mL. In some embodiments, the concentration of polysorbate 20 is about 0.01 mg / mL, about 0.02 mg / mL, about 0.05 mg / mL, about 0.1 mg / mL, about 0.2 mg / mL, about 0.3 mg / mL, about 0.4 mg / mL, about 0.5 mg / mL, about 0.6 mg / mL, about 0.7 mg / mL, about 0.8 mg / mL, about 0.9 mg / mL, about 1.0 mg / mL, about 1.5 mg / mL, or about 2.0 mg / mL. In some embodiments, the concentration of polysorbate 20 is about 0.1 mg / mL. In some embodiments, the concentration of polysorbate 20 is about 0.6 mg / mL. In some embodiments, the concentration of polysorbate 20 is 0.01 mg / mL, 0.02 mg / mL, 0.05 mg / mL, 0.1 mg / mL, 0.2 mg / mL, 0.3 mg / mL, 0.4 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, 1.5 mg / mL, or 2.0 mg / mL, or any range between these values. In some embodiments, the concentration of polysorbate 20 is 0.1 mg / mL. In some embodiments, the concentration of polysorbate 20 is 0.6 mg / mL.

[0039] antioxidants

[0040] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments further comprises an antioxidant. In some embodiments, the antioxidant is a metal ion chelating agent. In some embodiments, the antioxidant is ethylenediaminetetraacetic acid (EDTA) or a salt thereof, EDTA hydrate or a salt thereof. In some embodiments, the antioxidant is EDTA hydrate or a salt thereof. In some embodiments, the antioxidant is a sodium salt of EDTA hydrate. In some embodiments, the antioxidant is disodium edetate (EDTA-2Na).

[0041] In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of disodium edetate is from 0.01 mg / mL to 2.0 mg / mL. In some embodiments, the concentration of disodium edetate is from 0.01 mg / mL to 1.0 mg / mL. In some embodiments, the concentration of disodium edetate is from 0.01 mg / mL to 0.1 mg / mL. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of disodium edetate is from 0.01 mg / mL to 0.5 mg / mL. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of disodium edetate is from 0.05 mg / mL to 0.2 mg / mL. In some embodiments, the concentration of the disodium edetate is about 0.01 mg / mL, about 0.02 mg / mL, about 0.03 mg / mL, about 0.04 mg / mL, about 0.05 mg / mL, about 0.06 mg / mL, about 0.07 mg / mL, about 0.08 mg / mL, about 0.09 mg / mL, about 0.1 mg / mL, about 0.2 mg / mL, about 0.3 mg / mL, about 0.4 mg / mL, about 0.5 mg / mL, about 0.6 mg / mL, about 0.7 mg / mL, about 0.8 mg / mL, about 0.9 mg / mL, about 1.0 mg / mL, or about 2.0 mg / mL. In some embodiments, the concentration of the disodium edetate is about 0.1 mg / mL. In some embodiments, the concentration of the disodium edetate is 0.01 mg / mL, 0.02 mg / mL, 0.03 mg / mL, 0.04 mg / mL, 0.05 mg / mL, 0.06 mg / mL, 0.07 mg / mL, 0.08 mg / mL, 0.09 mg / mL, 0.1 mg / mL, 0.2 mg / mL, 0.3 mg / mL, 0.4 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, or 2.0 mg / mL, or any range between these values. In some embodiments, the concentration of the disodium edetate is 0.1 mg / mL.

[0042] In some embodiments, such as the pharmaceutical composition described in any of the preceding embodiments, the antioxidant is methionine.

[0043] In some embodiments, the concentration of methionine in the pharmaceutical composition as described in any of the preceding embodiments is from 1 mM to 200 mM. In some embodiments, the concentration of methionine is from 1 mM to 20 mM. In some embodiments, the concentration of methionine is from 5 mM to 15 mM. In some embodiments, the concentration of methionine is from 8 mM to 12 mM. In some embodiments, the concentration of methionine is from 9 mM to 11 mM. In some embodiments, the concentration of the methionine is about 1 mM, about 3 mM, about 5 mM, about 8 mM, about 9 mM, about 10 mM, about 11 mM, about 12 mM, about 15 mM, about 20 mM, about 25 mM, about 30 mM, about 50 mM, about 75 mM, about 80 mM, about 90 mM, about 100 mM, about 110 mM, about 120 mM, about 150 mM, about 160 mM, about 180 mM, or about 200 mM. In some embodiments, the concentration of the methionine is about 10 mM. In some embodiments, the concentration of methionine is 1 mM, 3 mM, 5 mM, 8 mM, 9 mM, 10 mM, 11 mM, 12 mM, 15 mM, 20 mM, 25 mM, 30 mM, 50 mM, 75 mM, 80 mM, 90 mM, 100 mM, 110 mM, 120 mM, 150 mM, 160 mM, 180 mM, or 200 mM, or any range between these values. In some embodiments, the concentration of methionine is 10 mM.

[0044] Hyaluronidase

[0045] In some embodiments, the pharmaceutical composition as described in any of the preceding claims further comprises hyaluronidase. In some embodiments, the hyaluronidase in the pharmaceutical composition as described in any of the preceding claims is recombinant human hyaluronidase (rHuPH20) or a variant thereof. The amino acid sequence of rHuPH20 (HYLENEXTM) is well known and can be obtained with CAS Registry No. 757971-58-7. Its approximate molecular weight is 61 kDa (see also U.S. Patent No. 7,767,429).

[0046] In some embodiments, the pharmaceutical composition as described in any of the preceding claims, the hyaluronidase comprises SEQ ID NO: 11, or an amino acid sequence having at least 80% (e.g., at least 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity with it. In some embodiments, the hyaluronidase comprises the amino acid sequence shown in SEQ ID NO: 11. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, the sequence of the hyaluronidase is as shown in SEQ ID NO: 11.

[0047] >PH20 (SEQ ID NO: 11)

[0048] In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the hyaluronidase activity is greater than 500 U / mL. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the hyaluronidase activity is from 1000 U / mL to 3000 U / mL. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the hyaluronidase activity is from 1600 U / mL to 2400 U / mL. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the hyaluronidase activity is from 1800 U / mL to 2200 U / mL. In some embodiments, the hyaluronidase activity is about 1000 U / mL, about 1100 U / mL, about 1200 U / mL, about 1300 U / mL, about 1400 U / mL, about 1500 U / mL, about 1600 U / mL, about 1700 U / mL, about 1800 U / mL, about 1900 U / mL, about 2000 U / mL, about 2100 U / mL, about 2200 U / mL, about 2300 U / mL, about 2400 U / mL, about 2500 U / mL, about 2600 U / mL, about 2700 U / mL, about 2800 U / mL, about 2900 U / mL, or about 3000 U / mL. In some embodiments, in the pharmaceutical composition as described in any of the preceding claims, the hyaluronidase activity is about 2000 U / mL. In some embodiments, the hyaluronidase activity is 1000 U / mL, 1100 U / mL, 1200 U / mL, 1300 U / mL, 1400 U / mL, 1500 U / mL, 1600 U / mL, 1700 U / mL, 1800 U / mL, 1900 U / mL, 2000 U / mL, 2100 U / mL, 2200 U / mL, 2300 U / mL, 2400 U / mL, 2500 U / mL, 2600 U / mL, 2700 U / mL, 2800 U / mL, 2900 U / mL, or 3000 U / mL, and any range between these values. In some embodiments, in the pharmaceutical composition as described in any of the preceding claims, the hyaluronidase activity is 2000 U / mL.

[0049] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments comprises the following components:

[0050] a) Anti-PD-L1 antibody at concentrations of 80 mg / mL to 180 mg / mL;

[0051] b) 5mM to 50mM histidine-histidine hydrochloride buffer;

[0052] c) Sucrose at concentrations of 50 mg / mL to 90 mg / mL;

[0053] d) Polysorbate at concentrations of 0.1 mg / mL to 1.0 mg / mL;

[0054] e) Disodium edetate at concentrations of 0.01 mg / mL to 1.0 mg / mL;

[0055] The pH of the pharmaceutical composition is 5.0 to 5.8.

[0056] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments comprises the following components:

[0057] a) Anti-PD-L1 antibody at concentrations of 80 mg / mL to 180 mg / mL;

[0058] b) 5mM to 50mM histidine-histidine hydrochloride buffer;

[0059] c) Sucrose at concentrations of 50 mg / mL to 90 mg / mL;

[0060] d) Polysorbate 80 at concentrations of 0.1 mg / mL to 1.0 mg / mL;

[0061] e) Disodium edetate at concentrations of 0.01 mg / mL to 1.0 mg / mL;

[0062] The pH of the pharmaceutical composition is 5.0 to 5.8.

[0063] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments comprises the following components:

[0064] a) Anti-PD-L1 antibody at concentrations of 80 mg / mL to 180 mg / mL;

[0065] b) 5mM to 50mM histidine-histidine hydrochloride buffer;

[0066] c) Sucrose at concentrations of 50 mg / mL to 90 mg / mL;

[0067] d) Polysorbate 80 at concentrations of 0.1 mg / mL to 1.0 mg / mL;

[0068] e) Disodium edetate at concentrations of 0.01 mg / mL to 1.0 mg / mL;

[0069] f) Methionine from 1 mM to 20 mM;

[0070] 1000 U / mL to 3000 U / mL of hyaluronidase;

[0071] The pH of the pharmaceutical composition is 5.0 to 5.8.

[0072] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments comprises the following components:

[0073] a) Anti-PD-L1 antibody at concentrations of 120 mg / mL to 180 mg / mL;

[0074] b) 10 mM to 40 mM histidine-histidine hydrochloride buffer;

[0075] c) Sucrose at concentrations of 55 mg / mL to 85 mg / mL;

[0076] d) Polysorbate 80 at concentrations of 0.1 mg / mL to 0.8 mg / mL;

[0077] e) Disodium edetate at concentrations of 0.01 mg / mL to 0.5 mg / mL;

[0078] f) Methionine from 5 mM to 15 mM;

[0079] 1600 U / mL to 2400 U / mL of hyaluronidase;

[0080] The pH of the pharmaceutical composition is 5.0 to 5.8.

[0081] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments comprises the following components:

[0082] a) Anti-PD-L1 antibody at concentrations of 135 mg / mL to 165 mg / mL;

[0083] b) 15mM to 30mM histidine-histidine hydrochloride buffer;

[0084] c) Sucrose at concentrations of 65 mg / mL to 75 mg / mL;

[0085] d) Polysorbate 80 at concentrations of 0.4 mg / mL to 0.8 mg / mL;

[0086] e) Disodium edetate at concentrations of 0.05 mg / mL to 0.2 mg / mL;

[0087] f) Methionine from 5 mM to 15 mM;

[0088] 1800 U / mL to 2200 U / mL of hyaluronidase;

[0089] The pH of the pharmaceutical composition is 5.0 to 5.5.

[0090] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments comprises the following components:

[0091] a) Anti-PD-L1 antibody at concentrations of 135 mg / mL to 165 mg / mL;

[0092] b) 15 mM to 25 mM histidine-histidine hydrochloride buffer;

[0093] c) Sucrose at concentrations of 65 mg / mL to 75 mg / mL;

[0094] d) Polysorbate 80 at concentrations of 0.4 mg / mL to 0.8 mg / mL;

[0095] e) Disodium edetate at concentrations of 0.05 mg / mL to 0.2 mg / mL;

[0096] f) Methionine from 5 mM to 15 mM;

[0097] 1800 U / mL to 2200 U / mL of hyaluronidase;

[0098] The pH of the pharmaceutical composition is 5.0 to 5.5.

[0099] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments comprises the following components:

[0100] a) Approximately 150 mg / mL of anti-PD-L1 antibody;

[0101] b) Approximately 20 mM histidine-histidine hydrochloride buffer;

[0102] c) Approximately 70 mg / mL of sucrose;

[0103] d) Approximately 0.6 mg / mL of polysorbate 80;

[0104] e) Approximately 0.1 mg / mL of disodium edetate;

[0105] f) Approximately 10 mM of methionine;

[0106] g) Approximately 2000 U / mL of hyaluronidase;

[0107] The pH of the pharmaceutical composition is approximately 5.3.

[0108] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments comprises the following components:

[0109] a) 150 mg / mL of anti-PD-L1 antibody;

[0110] b) 20 mM histidine-histidine hydrochloride buffer;

[0111] c) 70 mg / mL of sucrose;

[0112] d) 0.6 mg / mL polysorbate 80;

[0113] e) 0.1 mg / mL of disodium edetate;

[0114] f) 10 mM methionine;

[0115] 2000 U / mL hyaluronidase;

[0116] The pH of the pharmaceutical composition is 5.3.

[0117] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments comprises the following components:

[0118] a) Approximately 150 mg / mL of anti-PD-L1 antibody;

[0119] b) Approximately 30 mM histidine-histidine hydrochloride buffer;

[0120] c) Approximately 70 mg / mL of sucrose;

[0121] d) Approximately 0.6 mg / mL of polysorbate 80;

[0122] e) Approximately 0.1 mg / mL of disodium edetate;

[0123] f) Approximately 10 mM of methionine;

[0124] g) Approximately 2000 U / mL of hyaluronidase;

[0125] The pH of the pharmaceutical composition is approximately 5.2.

[0126] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments comprises the following components:

[0127] a) 150 mg / mL of anti-PD-L1 antibody;

[0128] b) 30 mM histidine-histidine hydrochloride buffer;

[0129] c) 70 mg / mL of sucrose;

[0130] d) 0.6 mg / mL polysorbate 80;

[0131] e) 0.1 mg / mL of disodium edetate;

[0132] f) 10 mM methionine;

[0133] 2000 U / mL hyaluronidase;

[0134] The pH of the pharmaceutical composition is 5.2.

[0135] In another aspect, this disclosure provides a pharmaceutical composition comprising:

[0136] a) Anti-PD-L1 antibody at concentrations of 80 mg / mL to 180 mg / mL;

[0137] b) 5mM to 50mM histidine-histidine hydrochloride buffer;

[0138] c) Sucrose at concentrations of 50 mg / mL to 90 mg / mL;

[0139] d) Polysorbate at concentrations of 0.1 mg / mL to 1.0 mg / mL;

[0140] e) Disodium edetate at concentrations of 0.01 mg / mL to 1.0 mg / mL;

[0141] Furthermore, the pH of the pharmaceutical composition is 5.0 to 5.8;

[0142] in:

[0143] The anti-PD-L1 antibody comprises a heavy chain variable region and a light chain variable region. The heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3. The HCDR1 of the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 1, the HCDR2 comprises the amino acid sequence of SEQ ID NO: 2, and the HCDR3 comprises the amino acid sequence of SEQ ID NO: 3. The LCDR1 of the light chain variable region comprises the amino acid sequence of SEQ ID NO: 4, the LCDR2 comprises the amino acid sequence of SEQ ID NO: 5, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 6.

[0144] In some embodiments, the pharmaceutical composition as described above further comprises methionine.

[0145] In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of methionine is from 1 mM to 20 mM. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of methionine is from 5 mM to 15 mM. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of methionine is about 10 mM.

[0146] In some embodiments, the pharmaceutical composition as described in any of the preceding claims further comprises hyaluronidase. In some embodiments, the hyaluronidase in the pharmaceutical composition as described in any of the preceding claims is recombinant human hyaluronidase (rHuPH20). In some embodiments, the hyaluronidase in the pharmaceutical composition as described in any of the preceding claims comprises the amino acid sequence shown in SEQ ID NO: 11.

[0147] In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of hyaluronidase is from 1000 U / mL to 3000 U / mL. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of hyaluronidase is from 1600 U / mL to 2400 U / mL. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of hyaluronidase is from 1800 U / mL to 2200 U / mL. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of hyaluronidase is about 2000 U / mL.

[0148] In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of the anti-PD-L1 antibody is from 120 mg / mL to 180 mg / mL. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of the anti-PD-L1 antibody is from 135 mg / mL to 165 mg / mL. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of the anti-PD-L1 antibody is about 150 mg / mL.

[0149] In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of the histidine-histidine hydrochloride buffer is from 10 mM to 40 mM. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of the histidine-histidine hydrochloride buffer is from 15 mM to 30 mM. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of the histidine-histidine hydrochloride buffer is about 20 mM or about 30 mM.

[0150] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments has a pH of 5.0 to 5.5. In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments has a pH of about 5.3 or about 5.2.

[0151] In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of sucrose is from 55 mg / mL to 85 mg / mL. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of sucrose is from 65 mg / mL to 75 mg / mL. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of sucrose is about 70 mg / mL.

[0152] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments, wherein the polysorbate is polysorbate 80 or polysorbate 20. In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments, wherein the polysorbate is polysorbate 80.

[0153] In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of polysorbate is from 0.1 mg / mL to 0.8 mg / mL. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of polysorbate is from 0.4 mg / mL to 0.8 mg / mL. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of polysorbate is about 0.6 mg / mL.

[0154] In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of disodium edetate is from 0.01 mg / mL to 0.5 mg / mL. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of disodium edetate is from 0.05 mg / mL to 0.2 mg / mL. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the concentration of disodium edetate is about 0.1 mg / mL.

[0155] In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the anti-PD-L1 antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 7, or an amino acid sequence having at least 80% sequence identity with it; and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 8, or an amino acid sequence having at least 80% sequence identity with it.

[0156] In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the anti-PD-L1 antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 7, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 8.

[0157] In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the anti-PD-L1 antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 9, or an amino acid sequence having at least 80% sequence identity with it; and the light chain comprises the amino acid sequence of SEQ ID NO: 10, or an amino acid sequence having at least 80% sequence identity with it.

[0158] In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the anti-PD-L1 antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 9, and the light chain comprises the amino acid sequence of SEQ ID NO: 10.

[0159] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments comprises the following components:

[0160] a) Anti-PD-L1 antibody at concentrations of 80 mg / mL to 180 mg / mL;

[0161] b) 5mM to 50mM histidine-histidine hydrochloride buffer;

[0162] c) Sucrose at concentrations of 50 mg / mL to 90 mg / mL;

[0163] d) Polysorbate 80 at concentrations of 0.1 mg / mL to 1.0 mg / mL;

[0164] e) Disodium edetate at concentrations of 0.01 mg / mL to 1.0 mg / mL;

[0165] f) Methionine from 1 mM to 20 mM;

[0166] 1000 U / mL to 3000 U / mL of hyaluronidase;

[0167] The pH of the pharmaceutical composition is 5.0 to 5.8.

[0168] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments comprises the following components:

[0169] a) Anti-PD-L1 antibody at concentrations of 120 mg / mL to 180 mg / mL;

[0170] b) 10 mM to 40 mM histidine-histidine hydrochloride buffer;

[0171] c) Sucrose at concentrations of 55 mg / mL to 85 mg / mL;

[0172] d) Polysorbate 80 at concentrations of 0.1 mg / mL to 0.8 mg / mL;

[0173] e) Disodium edetate at concentrations of 0.01 mg / mL to 0.5 mg / mL;

[0174] f) Methionine from 5 mM to 15 mM;

[0175] 1600 U / mL to 2400 U / mL of hyaluronidase;

[0176] The pH of the pharmaceutical composition is 5.0 to 5.8.

[0177] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments comprises the following components:

[0178] a) Anti-PD-L1 antibody at concentrations of 135 mg / mL to 165 mg / mL;

[0179] b) 15mM to 30mM histidine-histidine hydrochloride buffer;

[0180] c) Sucrose at concentrations of 65 mg / mL to 75 mg / mL;

[0181] d) Polysorbate 80 at concentrations of 0.4 mg / mL to 0.8 mg / mL;

[0182] e) Disodium edetate at concentrations of 0.05 mg / mL to 0.2 mg / mL;

[0183] f) Methionine from 5 mM to 15 mM;

[0184] 1800 U / mL to 2200 U / mL of hyaluronidase;

[0185] The pH of the pharmaceutical composition is 5.0 to 5.5.

[0186] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments comprises the following components:

[0187] a) Approximately 150 mg / mL of anti-PD-L1 antibody;

[0188] b) Approximately 20 mM histidine-histidine hydrochloride buffer;

[0189] c) Approximately 70 mg / mL of sucrose;

[0190] d) Approximately 0.6 mg / mL of polysorbate 80;

[0191] e) Approximately 0.1 mg / mL of disodium edetate;

[0192] f) Approximately 10 mM of methionine;

[0193] g) Approximately 2000 U / mL of hyaluronidase;

[0194] The pH of the pharmaceutical composition is approximately 5.3.

[0195] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments is a fixed combination or a non-fixed combination. In some embodiments, the pharmaceutical composition is a fixed combination.

[0196] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments is a liquid formulation. In some embodiments, the solvent of the liquid formulation is water.

[0197] In some embodiments, the pharmaceutical composition, as described in any of the preceding embodiments, is a subcutaneous injection formulation.

[0198] In some embodiments, the pharmaceutical composition, as described in any of the preceding embodiments, is suitable for subcutaneous injection.

[0199] In some embodiments, the pharmaceutical composition as described in any of the preceding claims is a drug for preparation of a subcutaneous injection. In some embodiments, the pharmaceutical composition as described in any of the preceding claims is stable at 2-8°C and can be stored for at least about 1 week, 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, at least about 2 years, or at least about 5 years. In some embodiments, the pharmaceutical composition as described in any of the preceding claims is stable at 2-8°C for at least 6 months. In some embodiments, the pharmaceutical composition as described in any of the preceding claims is stable at 2-8°C for at least 1 year. In some embodiments, the pharmaceutical composition as described in any of the preceding claims is stable at 2-8°C for at least 2 years.

[0200] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments is stable at about 5°C and can be stored for at least about 1 week, 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, at least about 2 years, or at least about 5 years. In some embodiments, the pharmaceutical composition is stable at about 40°C and can be stored for at least about 1 week, 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, at least about 2 years, or at least about 5 years. In other embodiments, the pharmaceutical composition is stable at about 25°C and can be stored for at least about 1 week, 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, at least about 2 years, or at least about 5 years.

[0201] In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the pharmaceutical composition exhibits minimal change in its acidic peak after prolonged storage under stress, such as at a specific temperature. In one embodiment, the pharmaceutical composition exhibits a change in its acidic peak of less than about 10%, about 9%, about 8%, about 7%, about 6%, about 5%, about 4%, about 3%, about 2%, about 1%, about 0.9%, about 0.8%, about 0.7%, about 0.6%, about 0.5%, about 0.4%, about 0.3%, about 0.2%, or about 0.1% after storage at 2-8°C for about 6 months. In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the pharmaceutical composition is stable at 2-8°C for 6 months and its IEC acidic peak change is about 0.1%.

[0202] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments exhibits minimal change in its basic peak after prolonged storage under stress, such as at a specific temperature. In one embodiment, the pharmaceutical composition exhibits a change in its basic peak of less than about 10%, about 9%, about 8%, about 7%, about 6%, about 5%, about 4%, about 3%, about 2%, or about 1.1% after storage at 2-8°C for about 6 months. In another embodiment, the pharmaceutical composition is stable at 2-8°C for 6 months, and its IEC basic peak changes by about 1.1%.

[0203] The pharmaceutical compositions disclosed herein may also exhibit minimal change in polymer peaks after prolonged storage, lasting weeks, months, or years. In one embodiment, after storage at 2-8°C for approximately 6 months, the pharmaceutical compositions exhibit changes in polymer peaks of less than approximately 5%, approximately 4%, approximately 3%, approximately 2%, approximately 1.5%, approximately 1.4%, approximately 1.3%, approximately 1.2%, approximately 1.1%, approximately 1%, approximately 0.9%, approximately 0.8%, approximately 0.7%, approximately 0.6%, approximately 0.5%, approximately 0.4%, approximately 0.3%, approximately 0.2%, or approximately 0.1%. In one embodiment, the pharmaceutical compositions are stable at 2-8°C for 6 months with a change in their polymer peaks of approximately 0.1%.

[0204] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments, wherein the anti-PD-L1 antibody in the pharmaceutical composition retains at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 105%, 110%, 115%, 120%, 125%, 130%, 135%, or 140%) of its biological activity (e.g., binding activity) after storage. In some embodiments, the biological activity is measured by binding to PD-L1.

[0205] In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the hyaluronidase in the pharmaceutical composition retains at least 60% (e.g., 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 105%, 110%, 115%, 120%, 125%, 130%, 135%, or 140%) of its biological activity (e.g., enzyme activity) after storage. In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments, wherein the hyaluronidase in the pharmaceutical composition retains at least 80% (e.g., at least 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) of its biological activity (e.g., enzyme activity) after storage. In some embodiments, the biological activity is measured by a multifunctional microplate reader.

[0206] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments, wherein the degradation rate of said polysorbate 80 is at least less than 50% (e.g., less than 45%, less than 40%, less than 35%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%).

[0207] In some embodiments, the pharmaceutical composition as described in any of the preceding claims has a viscosity not exceeding 20 cp (centipoise). In some embodiments, the viscosity of the pharmaceutical composition is from 2 cp to 20 cp. In some embodiments, the viscosity of the pharmaceutical composition is from 2 cp to 13 cp. In some embodiments, the pharmaceutical composition as described in any of the preceding claims has a viscosity of 2 cp to 13 cp when the concentration of the anti-PD-L1 antibody is from 80 mg / mL to 150 mg / mL. In some embodiments, the pharmaceutical composition as described in any of the preceding claims has a viscosity of 12 cp to 13 cp when the concentration of the anti-PD-L1 antibody is 150 mg / mL. In some embodiments, the viscosity of the pharmaceutical composition is measured at 25°C.

[0208] In another aspect, this disclosure also provides a method for preparing a lyophilized formulation, comprising the step of lyophilizing a pharmaceutical composition as described in any of the preceding claims.

[0209] In another aspect, this disclosure also provides a lyophilized formulation obtained by the method described above.

[0210] In another aspect, this disclosure also provides a lyophilized formulation comprising the pharmaceutical composition as described in any of the preceding claims.

[0211] In another aspect, this disclosure also provides a lyophilized formulation that, upon reconstitution, can form a pharmaceutical composition as described in any of the preceding claims.

[0212] In another aspect, this disclosure also provides a reconstituted solution, characterized in that the reconstituted solution is prepared by reconstituted a lyophilized formulation as described in any of the preceding claims.

[0213] In another aspect, this disclosure also provides a reconstituted solution, which is a reconstituted form of the lyophilized formulation as described in any of the preceding claims.

[0214] In some embodiments, the reconstituted solution as described in the preceding claim has the same components and contents as the pharmaceutical composition as described in the preceding claim.

[0215] In another aspect, this disclosure also provides an article of manufacture comprising a container containing a pharmaceutical composition as described in any of the preceding claims, or a lyophilized formulation as described in any of the preceding claims, or a reconstituted solution as described in any of the preceding claims.

[0216] In another aspect, this disclosure also provides a method for treating or preventing a disease, the method comprising administering to a subject a therapeutically effective amount of a pharmaceutical composition as described in any of the preceding claims, or a lyophilized formulation as described in any of the preceding claims, or a reconstituted solution as described in any of the preceding claims.

[0217] In another aspect, this disclosure also provides the use of the pharmaceutical compositions as described in any of the preceding claims, or the lyophilized formulations as described in any of the preceding claims, or the reconstituted solutions as described in any of the preceding claims, in the preparation of medicaments for the treatment or prevention of diseases.

[0218] In another aspect, this disclosure also provides pharmaceutical compositions as described in any of the preceding claims, or lyophilized formulations as described in any of the preceding claims, or reconstituted solutions as described in any of the preceding claims, said pharmaceuticals for the treatment or prevention of disease.

[0219] In some implementations, the disease is a tumor.

[0220] In some implementations, the disease is a solid tumor or a malignant tumor.

[0221] In some implementations, the disease is an advanced solid tumor or an advanced malignant tumor.

[0222] In some implementations, the disease is cancer.

[0223] In some implementations, the disease is a cancer that expresses PD-L1.

[0224] In some implementations, the disease is breast cancer, lung cancer (e.g., non-small cell lung cancer, small cell lung cancer, lung adenocarcinoma), gastric cancer (e.g., gastric / gastroesophageal junction adenocarcinoma), colorectal cancer (e.g., colorectal cancer, rectal adenocarcinoma), kidney cancer, melanoma, liver cancer (e.g., hepatocellular carcinoma), esophageal cancer (e.g., esophageal squamous cell carcinoma, esophageal squamous cell carcinoma, esophageal small cell carcinoma, esophageal cancer), cervical cancer, ovarian cancer, endometrial cancer, biliary tract cancer (e.g., biliary adenocarcinoma, bile duct cancer), pancreatic cancer, thyroid cancer, bladder cancer, nasopharyngeal carcinoma, prostate cancer, or squamous cell carcinoma (e.g., oral squamous cell carcinoma, head and neck squamous cell carcinoma). Detailed Implementation

[0225] the term

[0226] To facilitate understanding of this disclosure, certain technical and scientific terms are specifically defined below. Unless otherwise expressly defined herein, all other technical and scientific terms used herein have the meanings commonly understood by one of ordinary skill in the art to which this disclosure pertains.

[0227] The singular forms “a,” “an,” and “the” used in this disclosure include plural references unless the context clearly indicates otherwise.

[0228] Unless the context clearly requires otherwise, the words “comprising,” “having,” “including,” etc., in the patent specification and claims should be understood as “including but not limited to,” rather than as exclusive or exhaustive.

[0229] "Optional" or "optionally" means that the event or circumstances described below may, but do not have to, occur, including the circumstances in which the event or circumstances may or may not occur.

[0230] Those skilled in the art will understand that when used as a reference range, cutoff value, or specific value, "about" can mean within one or more standard deviations. Alternatively, "about" can mean a range with a difference of up to 20% (i.e., ±20%). Since many of the values ​​used herein were determined experimentally, those skilled in the art will understand that such determinations can vary between different experiments and are generally true across experiments. Due to this inherent variability, the values ​​used herein should not be unduly restricted. Therefore, the term "about" is used to cover variations of ±20%, ±10%, ±5%, ±1%, ±0.5%, or ±0.1% or less from a specified value.

[0231] Although this disclosure provides content ranges or content values, those skilled in the art will understand that the content ranges or content values ​​cover the acceptable range of error for the specific values ​​measured.

[0232] The three-letter and single-letter codes for amino acids used in this disclosure are as described in J. Biol. Chem., 243, p3558 (1968).

[0233] The term "amino acid" refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimics that function in a manner similar to naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code, as well as those that are subsequently modified, such as hydroxyproline, γ-carboxyglutamic acid, and O-phosphoserine. Amino acid analogs are compounds that have the same basic chemical structure as naturally occurring amino acids (i.e., the α-carbon bound to hydrogen, carboxyl, amino, and R groups), such as homoserine, ortholeucine, methionine sulfoxide, and methionine methylsulfonium. These analogs have modified R groups (e.g., ortholeucine) or modified peptide backbones but retain the same basic chemical structure as naturally occurring amino acids. Amino acid mimics are chemical compounds that have a structure different from the general chemical structure of amino acids but function in a manner similar to naturally occurring amino acids.

[0234] The term "antibody" is used in the broadest sense and covers a wide range of antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies; monospecific antibodies, multispecific antibodies (e.g., bispecific antibodies); full-length antibodies and antigen-binding fragments (or antigen-binding portions), as long as they exhibit the desired antigen-binding activity.

[0235] "Natural antibodies" refer to naturally occurring immunoglobulin molecules. For example, natural IgG antibodies are heterotetraglycoproteins of approximately 150,000 Daltons, composed of two identical light chains and two identical heavy chains linked by disulfide bonds. From the N to the C-terminus, each heavy chain has a variable region (VH), also known as a variable heavy domain or heavy chain variable region, followed by a heavy chain constant region. The IgG heavy chain constant region (CH) typically contains three constant regions (CH1, CH2, and CH3). Similarly, from the N to the C-terminus, each light chain has a variable region (VL), also known as a variable light domain or light chain variable domain, followed by a constant light domain (light chain constant region, CL).

[0236] The term "variable region" or "variable domain" refers to the domain in the antibody heavy or light chain involved in antibody-antigen binding. In this paper, the antibody heavy chain variable region (VH) and light chain variable region (VL) each contain four conserved frame regions (FRs) and three complementarity-determining regions (CDRs). The term "complementarity-determining region" or "CDR" refers to the region within the variable domain that primarily facilitates antigen binding; "frame" or "FR" refers to the variable domain residues other than the CDR residues. The VH contains three CDR regions: HCDR1, HCDR2, and HCDR3; the VL contains three CDR regions: LCDR1, LCDR2, and LCDR3. Each VH and VL consists of three CDRs and four FRs arranged in the following order from the amino terminus (also called the N-terminus) to the carboxyl terminus (also called the C-terminus): FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. A single VH or VL may be sufficient to confer antigen-binding specificity.

[0237] The amino acid sequence boundaries of CDRs can be determined using various well-known schemes, such as the "Kabat" numbering rule, the "Chothia" numbering rule, the "ABM" numbering rule, the "contact" numbering rule, and the ImMunoGenTics (IMGT) numbering rule. The correspondence between various numbering systems is well known to those skilled in the art and is exemplified as shown in Table 1 below.

[0238] Table 1. Relationship between CDR numbering systems

[0239] Unless otherwise stated, the variable regions and CDR sequences in this disclosure embodiment are governed by the "Kabat" numbering rule. Although the Kabat numbering rule is used to define amino acid residues in specific implementations, corresponding technical solutions using other numbering systems are considered equivalent.

[0240] The term "monoclonal antibody" refers to a group of substantially homogeneous antibodies, meaning that the antibody molecules contained in this group have the same amino acid sequence, except for the possible small number of naturally occurring mutations. In contrast, polyclonal antibody formulations typically contain multiple different antibodies with different amino acid sequences in their variable structural domains, and they generally specifically target different epitopes. "Monoclonal" indicates the characteristic of an antibody obtained from a substantially homogeneous group of antibodies and should not be construed as requiring the antibody to be produced by any particular method. In some embodiments, the antibodies provided in this disclosure are monoclonal antibodies.

[0241] The antibodies disclosed herein may be derived from animals (such as antibodies from mice, birds, rabbits, camels, monkeys, etc.), chimeric antibodies, or humanized antibodies.

[0242] The term "chimeric" antibody refers to an antibody in which a portion of the heavy and / or light chain is derived from a specific source or species, while the remaining portion of the heavy and / or light chain is derived from another different source or species.

[0243] The term "humanized" antibody refers to an antibody that retains the reactivity of a non-human antibody while exhibiting lower immunogenicity in humans. For example, this can be achieved by retaining the non-human CDR region and replacing the rest of the antibody with its human counterpart (i.e., the frame region portion of the constant region and the variable region).

[0244] The term "affinity" refers to the overall strength of the non-covalent interaction between a single binding site of a molecule (e.g., an antibody) and its binding ligand (e.g., an antigen). Unless otherwise specified, as used herein, binding "affinity" refers to internal binding affinity, which reflects a 1:1 interaction between members of a binding pair (e.g., antibody and antigen). The affinity of molecule X for its ligand Y can typically be represented by the dissociation constant (KD). Affinity can be measured using conventional methods known in the art.

[0245] The terms “anti-PD-L1 antibody” and “PD-L1-binding antibody” refer to antibodies that can bind to PD-L1 or its epitopes with sufficient affinity.

[0246] The term "antigen" refers to a molecule or molecular moiety that can be bound by a selective binder of an antigen-binding protein (such as an antibody). An antigen may have one or more epitopes that can interact with different antigen-binding proteins (such as antibodies).

[0247] The term "epitope" refers to a region on an antigen that can specifically bind to an antibody or its antigen-binding fragment. Epitopes can be formed from consecutive amino acids (linear epitopes) or contain non-consecutive amino acids (conformal epitopes), for example, due to the folding of the antigen (i.e., tertiary folding of the antigen as a protein), which allows non-consecutive amino acids to be spatially close. The difference between conformational and linear epitopes is that antibody binding to a conformational epitope is lost in the presence of a denaturing solvent. An epitope contains at least 3, at least 4, at least 5, at least 6, at least 7, or 8-10 amino acids in a unique spatial conformation. Screening for antibodies that bind to a specific epitope (i.e., those that bind the same epitope) can be performed using methods routine in the art, such as, but not limited to, alanine scanning, Western blotting, peptide cleavage analysis, epitope excision, epitope extraction, chemical modification of the antigen, and cross-blocking.

[0248] The terms "capable of specific binding," "specific binding," or "binding" refer to the ability of an antibody to bind to a specific antigen or epitope with a higher affinity than other antigens or epitopes. Typically, antibodies bind at an affinity of approximately 1 × 10⁻⁶. -7An equilibrium dissociation constant (KD) of M or less binds to an antigen or epitope. In some embodiments, the KD of antibody binding to an antigen is 10% or less (e.g., 1%) of the KD of the antibody binding to a nonspecific antigen (e.g., BSA, casein). KD can be measured using known methods, such as by FACS or surface plasmon resonance assays. However, antibodies that specifically bind to an antigen or an epitope within an antigen may be cross-reactive to other related antigens, for example, to corresponding antigens from other species (homologous), such as humans or monkeys, such as the cynomolgus (cyno), the chimpanzee (chimp), or the common marmoset (marmoset).

[0249] "Surfactant" refers to a surface-active agent, preferably a nonionic surfactant. Surfactants can reduce protein aggregation and / or particle formation in formulations. The amount of surfactant added is such that it reduces protein aggregation and minimizes particle formation in the formulation.

[0250] "Disodium edetate" (CAS No.: 6381-92-6, the disodium edetate described in this disclosure was purchased from SIGMA). The company's website lists its alternative names as disodium edetate dihydrate, EDTA disodium salt, EDTA-Na2, EDTA Na2, disodium edetate dihydrate, disodium edetate dihydrate, and calcium sodium edetate. Therefore, in this disclosure, "disodium edetate," "disodium edetate dihydrate," and "EDTA-2Na" are interchangeable, as they all refer to the substance with CAS No.: 6381-92-6.

[0251] "Buffer solution" refers to a buffering agent that is resistant to pH changes through the action of its acid-base conjugate components. Examples of buffer solutions that maintain the pH within an appropriate range include acetate, succinate, gluconate, histidine, oxalate, lactate, phosphate, citrate (also known as tartrate), tartrate, fumarate, glycylglycine, and other organic acid buffer solutions.

[0252] "Histidine buffer" is a buffer solution containing histidine. Examples of histidine buffer solutions include histidine-histidine hydrochloride, histidine-histidine acetate, histidine-histidine phosphate, and histidine-histidine sulfate buffers, with histidine-histidine hydrochloride buffer being preferred. Histidine-histidine hydrochloride buffer can be prepared from histidine and hydrochloric acid, or from histidine and histidine hydrochloride.

[0253] "Succinate buffer" is a buffer solution containing succinate ions. Examples of succinate buffer solutions include sodium succinate, potassium succinate, and calcium succinate. A preferred succinate buffer solution is sodium succinate. Exemplarily, the sodium succinate may be prepared from succinic acid and sodium hydroxide, or from succinic acid and sodium succinate.

[0254] "Displacement" refers to the replacement of the solvent system in which antibody proteins are dissolved. For example, a buffer system containing antibody proteins may be physically replaced by a high-salt or hypertonic solvent system containing antibody proteins, thereby ensuring the presence of antibody proteins within the stable formulation. Such physical operations include, but are not limited to, ultrafiltration, dialysis, or centrifugation.

[0255] "Pharmaceutical composition" means a mixture containing one or more antibodies described herein or their physiologically / pharmacologically acceptable salts or prodrugs, along with other chemical components, such as physiologically / pharmacologically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to a living organism, thereby promoting the absorption of the active ingredient and the exertment of its biological activity.

[0256] The term "pharmaceutically acceptable salt" or "medicinal salt" refers to the salts of the antibody-drug conjugates disclosed herein, which are safe and effective when used in subjects and possess the intended biological activity. As an example, the antibody-drug conjugates disclosed herein contain at least one amino group and can therefore form salts with acids. Non-limiting examples of pharmaceutically acceptable salts include: hydrochloride, hydrobromide, hydroiodide, sulfate, hydrogen sulfate, citrate, acetate, succinate, ascorbate, oxalate, nitrate, sorbate, hydrogen phosphate, dihydrogen phosphate, salicylate, hydrogen citrate, tartrate, maleate, fumarate, formate, benzoate, methanesulfonate, ethanesulfonate, benzenesulfonate, and p-toluenesulfonate.

[0257] "Pharmaceutically acceptable carriers" or "pharmaceutically acceptable excipients" include any material that, when combined with an active ingredient, allows the ingredient to retain its biological activity and does not react with the subject's immune system. Examples include, but are not limited to, any standard pharmaceutical carrier, such as phosphate-buffered saline solutions, water, emulsions such as oil / water emulsions, and various types of wetting agents. In some embodiments, the diluent for aerosol or parenteral administration is phosphate-buffered saline (PBS) or physiological (0.9%) saline. Compositions containing such carriers are formulated using well-known conventional methods (see, for example, Remington's Pharmaceutical Sciences, 18th edition, A. Gennaro, editor, Mack Publishing Co., Easton, PA, 1990; and R. Remington, The Science and Practice of Pharmacy, 20th edition, Mack Publishing, 2000).

[0258] Unless otherwise specified, the solvent in the solution form of the pharmaceutical compositions described in this disclosure is water.

[0259] In this disclosure, "pharmaceutical composition" and "formulation" are not mutually exclusive.

[0260] The pharmaceutical compositions disclosed herein achieve a stable effect: the antibodies contained therein substantially retain their physical and / or chemical stability and / or biological activity after storage. Preferably, the pharmaceutical compositions substantially retain their physical and chemical stability and their biological activity after storage. The storage period is generally selected based on the intended shelf life of the pharmaceutical composition. Currently, there are various analytical techniques available for measuring protein stability, which can measure stability after storage at a selected temperature for a selected period of time.

[0261] Stable formulations include those in which no significant changes are observed when stored at refrigerated temperatures (2°C–8°C) for at least 1 month, at least 3 months, at least 6 months, preferably 1 year, and even more preferably up to 2 years. Stable liquid formulations also include those that exhibit the desired characteristics after storage at temperatures including 25°C for periods of 1 month, 3 months, or 6 months. Furthermore, stable liquid formulations also include those that exhibit the desired characteristics after storage at 40°C for periods of 4 weeks, 1 month, 3 months, or 6 months. Typical examples of stability include antibody aggregation or degradation typically not exceeding about 10%, preferably not exceeding about 5%, as determined by SEC-HPLC. Visually, the formulation is a pale yellow, nearly colorless, clear liquid or a colorless, clear liquid, or clear to slightly milky white. The concentration, pH, weight, and molecular osmotic pressure of the formulation exhibit aggregation of not more than about 10%, preferably not more than about 5%.

[0262] If, after visual inspection of color and / or clarity, or by means of UV light scattering, size exclusion chromatography (SEC), and dynamic light scattering (DLS), the antibody does not show significant increase in aggregation, precipitation, and / or denaturation, then the antibody “retains its physical stability” in the pharmaceutical formulation. Changes in protein conformation can be evaluated by fluorescence spectroscopy (which determines the tertiary structure of the protein) and by FTIR spectroscopy (which determines the secondary structure of the protein).

[0263] If an antibody does not exhibit significant chemical changes, then the antibody "retains its chemical stability" in the pharmaceutical formulation. Chemical stability can be assessed by detecting and quantifying the chemically altered form of the protein. Degradation processes that frequently alter the chemical structure of a protein include hydrolysis or truncation (evaluated by methods such as size exclusion chromatography and CE-SDS), oxidation (evaluated by methods such as peptide mapping combined with mass spectrometry or MALDI / TOF / MS), deamidation (evaluated by methods such as ion exchange chromatography, capillary isoelectric focusing, peptide mapping, and isofpartate measurement), and isomerization (evaluated by measuring isofpartate content, peptide mapping, etc.).

[0264] If the antibody's biological activity at a given time is within a predetermined range of the biological activity exhibited when the pharmaceutical formulation is prepared, then the antibody "retains its biological activity" in the pharmaceutical formulation.

[0265] "Administration," "giving," and "treatment," when applied to animals, humans, experimental subjects, cells, tissues, organs, or biological fluids, refer to the contact of an exogenous drug, therapeutic agent, diagnostic agent, or composition with the animal, human, subject, cell, tissue, organ, or biological fluid. "Administration," "giving," and "treatment" can refer to, for example, therapeutic, pharmacokinetic, diagnostic, research, and experimental methods. Cellular treatment includes contact between a reagent and cells, as well as contact between a reagent and a fluid, wherein the fluid is in contact with the cells. "Administration," "giving," and "treatment" also mean, by means of a reagent, diagnostic agent, conjugate composition, or by means of another cell in vitro and ex vivo, such as cells. "Treatment," when applied to humans, veterinary, or research subjects, refers to therapeutic treatment, preventative or prophylactic measures, research, and diagnostic applications.

[0266] "Treatment" means administering an oral or topical therapeutic agent, such as a pharmaceutical composition comprising any of the substances disclosed herein, to a patient who has symptoms of one or more diseases, and the therapeutic agent is known to have a therapeutic effect on these symptoms. Typically, a therapeutic agent is administered in a treated patient or population in an amount that effectively relieves symptoms of one or more diseases, in order to induce the regression of such symptoms or inhibit their progression to any clinically measurable degree. The amount of a therapeutic agent that effectively relieves any specific disease symptom (also referred to as a "therapeuticly effective amount") can vary depending on a variety of factors, such as the patient's disease state, age, and weight, and the drug's ability to produce the desired therapeutic effect in the patient. Whether the disease symptoms have been relieved can be evaluated using any clinical test method commonly used by a physician or other healthcare professional to assess the severity or progression of the symptoms. Although the embodiments disclosed herein (e.g., treatment methods or products) may be ineffective in alleviating symptoms of each target disease, they should reduce symptoms of the target disease in a statistically significant number of patients, as determined by any statistical test known in the art, such as the Student t-test, chi-square test, U-test according to Mann and Whitney, Kruskal-Wallis test (H-test), Jonckheere-Terpstra test, and Wilcoxon test.

[0267] "Effective amount" includes an amount sufficient to improve or prevent the symptoms or condition of a medical condition. Effective amount also means an amount sufficient to allow or facilitate a diagnosis. The effective amount used on a subject may vary depending on factors such as the condition to be treated, the subject's overall health, the route and dosage of administration, and the severity of side effects. Effective amount may be the maximum dose or administration regimen that avoids significant side effects or toxicity. Subjects disclosed herein may be animal or human subjects.

[0268] This document considers various dosing regimens, including but not limited to single or multiple administrations at multiple time points, bolus administration, and pulsatile infusion. In some embodiments, the pharmaceutical compositions disclosed herein are administered by subcutaneous injection.

[0269] The pharmaceutical compositions disclosed herein will be formulated, administered, and applied in accordance with good medical practice. Factors considered in this context include the specific condition being treated, the specific mammal being treated, the individual patient's clinical condition, the cause of the condition, the site of delivery of the agent, the method of administration, the timing of administration, and other factors known to a medical practitioner. Optionally, the pharmaceutical composition may also be formulated with one or more other agents for the prevention or treatment of said condition. The effective amount of such other agents depends on the amount of antigen-binding molecules present in the pharmaceutical composition, the type of condition or treatment, and other factors. It may be used at the same dosage and route of administration as described herein, or at about 1% to 99% of the dosage described herein, or at any dosage and any route of administration determined empirically / clinically to be appropriate.

[0270] The pharmaceutical composition containing an anti-PD-L1 antibody disclosed herein

[0271] For example, this disclosure provides a pharmaceutical composition comprising:

[0272] a) Anti-PD-L1 antibody at concentrations of 80 mg / mL to 180 mg / mL;

[0273] b) 5mM to 50mM histidine-histidine hydrochloride buffer;

[0274] c) Sucrose at concentrations of 50 mg / mL to 90 mg / mL;

[0275] d) Polysorbate at concentrations of 0.1 mg / mL to 1.0 mg / mL;

[0276] e) Disodium edetate at concentrations of 0.01 mg / mL to 1.0 mg / mL;

[0277] Furthermore, the pH of the pharmaceutical composition is 5.0 to 5.8;

[0278] in:

[0279] The anti-PD-L1 antibody comprises a heavy chain variable region and a light chain variable region. The heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3. The amino acid sequences of HCDR1 in the heavy chain variable region are shown in SEQ ID NO: 1, HCDR2 in SEQ ID NO: 2, and HCDR3 in SEQ ID NO: 3. The amino acid sequences of LCDR1 in the light chain variable region are shown in SEQ ID NO: 4, LCDR2 in SEQ ID NO: 5, and LCDR3 in SEQ ID NO: 6.

[0280] In some embodiments, the pharmaceutical composition as described in the preceding one further comprises methionine.

[0281] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments, wherein the concentration of methionine is from 1 mM to 20 mM. In some embodiments, the concentration of methionine is from 5 mM to 15 mM. In some embodiments, the concentration of methionine is about 10 mM.

[0282] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments further comprises hyaluronidase. In some embodiments, the hyaluronidase is recombinant human hyaluronidase (rHuPH20). In some embodiments, the sequence of the hyaluronidase is shown in SEQ ID NO: 11.

[0283] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments, wherein the hyaluronidase activity is from 1000 U / mL to 3000 U / mL. In some embodiments, the hyaluronidase activity is from 1600 U / mL to 2400 U / mL. In some embodiments, the hyaluronidase activity is from 1800 U / mL to 2200 U / mL. In some embodiments, the hyaluronidase activity is about 2000 U / mL.

[0284] In some embodiments, the pharmaceutical composition as described above, wherein the concentration of the anti-PD-L1 antibody is from 120 mg / mL to 180 mg / mL. In some embodiments, the concentration of the anti-PD-L1 antibody is from 135 mg / mL to 165 mg / mL. In some embodiments, the concentration of the anti-PD-L1 antibody is about 150 mg / mL.

[0285] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments, wherein the concentration of the histidine-histidine hydrochloride buffer is from 10 mM to 40 mM. In some embodiments, the concentration of the histidine-histidine hydrochloride buffer is from 15 mM to 30 mM. In some embodiments, the concentration of the histidine-histidine hydrochloride buffer is about 20 mM or about 30 mM.

[0286] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments has a pH of 5.0 to 5.5. In some embodiments, the pharmaceutical composition has a pH of about 5.3 or about 5.2.

[0287] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments, wherein the concentration of sucrose is from 55 mg / mL to 85 mg / mL. In some embodiments, the concentration of sucrose is from 65 mg / mL to 75 mg / mL. In some embodiments, the concentration of sucrose is about 70 mg / mL.

[0288] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments, wherein the polysorbate is polysorbate 80 or polysorbate 20. In some embodiments, the polysorbate is polysorbate 80.

[0289] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments, wherein the concentration of said polysorbate is from 0.1 mg / mL to 0.8 mg / mL. In some embodiments, the concentration of said polysorbate is from 0.4 mg / mL to 0.8 mg / mL. In some embodiments, the concentration of said polysorbate is about 0.6 mg / mL.

[0290] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments, wherein the concentration of disodium edetate is from 0.01 mg / mL to 0.5 mg / mL. In some embodiments, the concentration of disodium edetate is from 0.05 mg / mL to 0.2 mg / mL. In some embodiments, the concentration of disodium edetate is about 0.1 mg / mL.

[0291] In some embodiments, the anti-PD-L1 antibody comprises a heavy chain variable region and a light chain variable region, wherein 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.

[0292] In some embodiments, the pharmaceutical composition as described in any of the preceding claims, wherein the anti-PD-L1 antibody comprises a heavy chain and a light chain, wherein the amino acid sequence of the heavy chain is as shown in SEQ ID NO: 9, and the amino acid sequence of the light chain is as shown in SEQ ID NO: 10.

[0293] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments comprises the following components:

[0294] a) Anti-PD-L1 antibody at concentrations of 80 mg / mL to 180 mg / mL;

[0295] b) 5mM to 50mM histidine-histidine hydrochloride buffer;

[0296] c) Sucrose at concentrations of 50 mg / mL to 90 mg / mL;

[0297] d) Polysorbate 80 at concentrations of 0.1 mg / mL to 1.0 mg / mL;

[0298] e) Disodium edetate at concentrations of 0.01 mg / mL to 1.0 mg / mL;

[0299] f) Methionine from 1 mM to 20 mM;

[0300] 1000 U / mL to 3000 U / mL of hyaluronidase;

[0301] The pH of the pharmaceutical composition is 5.0 to 5.8.

[0302] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments comprises the following components:

[0303] a) Anti-PD-L1 antibody at concentrations of 120 mg / mL to 180 mg / mL;

[0304] b) 10 mM to 40 mM histidine-histidine hydrochloride buffer;

[0305] c) Sucrose at concentrations of 55 mg / mL to 85 mg / mL;

[0306] d) Polysorbate 80 at concentrations of 0.1 mg / mL to 0.8 mg / mL;

[0307] e) Disodium edetate at concentrations of 0.01 mg / mL to 0.5 mg / mL;

[0308] f) Methionine from 5 mM to 15 mM;

[0309] 1600 U / mL to 2400 U / mL of hyaluronidase;

[0310] The pH of the pharmaceutical composition is 5.0 to 5.8.

[0311] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments comprises the following components:

[0312] a) Anti-PD-L1 antibody at concentrations of 135 mg / mL to 165 mg / mL;

[0313] b) 15mM to 30mM histidine-histidine hydrochloride buffer;

[0314] c) Sucrose at concentrations of 65 mg / mL to 75 mg / mL;

[0315] d) Polysorbate 80 at concentrations of 0.4 mg / mL to 0.8 mg / mL;

[0316] e) Disodium edetate at concentrations of 0.05 mg / mL to 0.2 mg / mL;

[0317] f) Methionine from 5 mM to 15 mM;

[0318] 1800 U / mL to 2200 U / mL of hyaluronidase;

[0319] The pH of the pharmaceutical composition is 5.0 to 5.5.

[0320] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments comprises the following components:

[0321] a) Approximately 150 mg / mL of anti-PD-L1 antibody;

[0322] b) Approximately 20 mM histidine-histidine hydrochloride buffer;

[0323] c) Approximately 70 mg / mL of sucrose;

[0324] d) Approximately 0.6 mg / mL of polysorbate 80;

[0325] e) Approximately 0.1 mg / mL of disodium edetate;

[0326] f) Approximately 10 mM of methionine;

[0327] g) Approximately 2000 U / mL of hyaluronidase;

[0328] The pH of the pharmaceutical composition is approximately 5.3.

[0329] In some embodiments, the pharmaceutical composition as described in any of the preceding embodiments is a fixed combination or a non-fixed combination. In some embodiments, the pharmaceutical composition is a fixed combination.

[0330] In some embodiments, the pharmaceutical composition, as described in any of the preceding embodiments, is a liquid formulation.

[0331] In some embodiments, the pharmaceutical composition, as described in any of the preceding embodiments, is a subcutaneous injection formulation.

[0332] In some embodiments, the pharmaceutical composition, as described in any of the preceding embodiments, is suitable for subcutaneous injection.

[0333] In some embodiments, the pharmaceutical composition described in any of the preceding embodiments is used to prepare a drug for subcutaneous injection.

[0334] Details of one or more embodiments of this disclosure are set forth in the foregoing description. While any methods and materials similar to or the same as those described herein may be used to implement or test this disclosure, preferred methods and materials are described below. Other features, objects, and advantages of this disclosure will be apparent from the description and claims. In the description and claims, the singular form includes plural references unless the context clearly indicates otherwise. Unless otherwise defined, all technical and scientific terms used herein have their general meaning as understood by one of ordinary skill in the art to which this disclosure pertains. All patents and publications referenced in the description are incorporated herein by reference. The following embodiments are presented to illustrate preferred embodiments of this disclosure more fully. These embodiments should not be construed in any way as limiting the scope of this disclosure, which is defined by the claims.

[0335] Example

[0336] The present disclosure is further described below with reference to embodiments, but these embodiments are not intended to limit the scope of the invention.

[0337] Experimental methods not specifying specific conditions in the embodiments disclosed herein are generally performed under standard conditions or as recommended by the raw material or product manufacturer. Reagents not specifying their source are commercially available, standard reagents.

[0338] Example 1. Anti-PD-L1 antibody

[0339] WO2017084495A1 has disclosed the anti-PD-L1 antibody HRP00052, and the amino acid sequences of the heavy chain CDR and light chain CDR of the anti-PD-L1 antibody are shown in Table 2.

[0340] Table 2. CDR region sequences of the heavy and light chains of anti-PD-L1 antibodies Note: The CDRs in the table are CDRs determined according to the Kabat numbering system.

[0341] The heavy chain variable region and light chain variable region, and the heavy chain and light chain sequences of the anti-PD-L1 antibody are as follows: Anti-PD-L1 antibody heavy chain variable region

[0342] Anti-PD-L1 antibody light chain variable region Note: Underlined sequences are CDR sequences.

[0343] Anti-PD-L1 antibody heavy chain sequence

[0344] Anti-PD-L1 antibody light chain sequence Note: The underlined portion is the variable region sequence of the antibody heavy or light chain; the ununderlined portion is the constant region sequence of the antibody.

[0345] Example 2. Anti-PD-L1 antibody preparation

[0346] SEC size exclusion chromatography:

[0347] An analytical method for separating solutes based on the relative relationship between the pore size of the gel and the coil size of the polymer sample molecules.

[0348] SEC% polymer (SEC polymer content percentage) = A polymer / A total × 100% (A polymer is the peak area of ​​the polymer peak in the sample, and A total is the sum of the peak areas of all peaks).

[0349] ΔSEC% polymer = SEC% polymer of the formulation after stability testing - SEC% polymer of the formulation before stability testing.

[0350] Instrument used for SEC determination: Agilent HPLC 1260;

[0351] Pillars: Waters, BioResolve TM SEC mAb 2.5μm 7.8×300mm Column.

[0352] NR-CE capillary gel electrophoresis:

[0353] An electrophoresis method in which a gel is transferred into a capillary as a supporting medium and separated according to the molecular weight of the sample under a certain voltage.

[0354] NR-CE% fragments (percentage of NR-CE fragment content) = A fragments / A total × 100% (A fragments is the peak area of ​​fragment peaks in the sample, and A total is the sum of the peak areas of all peaks).

[0355] ΔNR-CE% fragments = NR-CE% fragments of the formulation after stability testing - NR-CE% fragments of the formulation before stability testing.

[0356] The instrument used for CE determination was a SCIEX capillary electrophoresis apparatus, model PA800 plus.

[0357] IEC ion exchange chromatography:

[0358] A chromatographic method that uses ion exchange resins or chemically bonded ion exchangers as the stationary phase and achieves separation by utilizing the differences in the ion exchange capacity or selectivity coefficients of the components to be separated.

[0359] IEC% acid peak (IEC acid peak content percentage) = A acid peak / A total × 100% (A acid peak is the peak area of ​​the acid peak in the sample, and A total area is the sum of the areas of the acid peak, neutral peak and basic peak).

[0360] IEC% alkaline peak (IEC alkaline peak content percentage) = A alkaline peak / A total × 100% (A alkaline peak is the peak area of ​​the alkaline peak in the sample, and A total area is the sum of the areas of the acidic peak, neutral peak and alkaline peak).

[0361] ΔIEC% acid peak = IEC% acid peak of the formulation after stability test - IEC% acid peak of the formulation before stability test.

[0362] ΔIEC% alkaline peak = IEC% alkaline peak of the formulation after stability test - IEC% alkaline peak of the formulation before stability test.

[0363] IEC measuring instrument: Agilent HPLC 1260.

[0364] ELSD anion exchange and reverse adsorption chromatography:

[0365] A method for quantifying polysorbate 80 is developed based on the principles of anion exchange and reverse adsorption to separate the solute and analyze it using electro-fogging detection technology.

[0366] PS80 degradation rate % = ((PS80 concentration of the formulation before stability test mg / ml - PS80 concentration of the formulation after stability test mg / ml) / PS80 concentration of the formulation before stability test mg / ml) × 100%.

[0367] ELSD determination instrument: Agilent HPLC 1260.

[0368] Enzyme activity assay:

[0369] Sodium hyaluronate was used as the substrate solution. The substrate solution before and after enzymatic hydrolysis was reacted with acidic bovine serum albumin, and the turbidity changes before and after enzymatic hydrolysis were compared to determine the activity of hyaluronidase.

[0370] Hyaluronidase activity remaining percentage % = Hyaluronidase activity of the formulation after stability test / Hyaluronidase activity of the formulation before stability test × 100.

[0371] Instrument used for enzyme activity assay: Multifunctional microplate reader, model MD Spectramax M5.

[0372] Viscosity measurement:

[0373] Viscosity is determined by a pressure-driven method based on Hagenpoisson's derivative equation, which measures viscosity by measuring shear stress and shear rate.

[0374] Viscosity measuring instrument: Unchained, model Honeybun.

[0375] Osmotic pressure measurement:

[0376] The freezing point method for determining osmotic pressure is based on the principle that the freezing point depression is directly proportional to the molar concentration of the solution. It uses a highly sensitive temperature sensing element to measure the freezing point of the solution and converts the electrical charge into osmotic pressure.

[0377] Instrument used for osmotic pressure measurement: Loser, model OM815.

[0378] protein:

[0379] The protein used in the following examples is the anti-PD-L1 antibody (hereinafter referred to as "protein") as described above.

[0380] The instrument used for protein concentration determination was a UV-Vis spectrophotometer, model: Nano Drop oneC, with a path length of 1 mm.

[0381] Example 2.1: Buffer System Screening

[0382] The buffer system shown in Table 3 was prepared, along with a formulation containing 150 mg / mL protein, 70 mg / mL sucrose, 0.6 mg / mL polysorbate 80 (PS80), 0.1 mg / mL disodium edetate (hereinafter referred to as "EDTA-2Na"), 10 mM methionine (Met), and 2000 U / mL hyaluronidase (PH20). Forced degradation studies (40℃ for 4 weeks) were conducted on the samples, and the effects of different buffer systems on protein stability were investigated using SEC, NR-CE, and IEC as evaluation indicators.

[0383] The results are shown in Table 3. After being placed at 40℃ for 4 weeks, there were no significant differences in the changes of SEC polymer peak and NR-CE fragment peak in the formulations of each buffer system. Among them, the changes of SEC polymer peak and NR-CE fragment peak in the His-HCl pH5.3 group were slightly lower, while the changes of IEC acid peak and alkaline peak were significantly lower than those in the His-AA pH5.3 group and SA pH5.3 group. Therefore, the formulation prepared by the His-HCl buffer system has the best stability.

[0384] Table 3. Results of buffer system screening Note: His-HCl represents histidine-histidine hydrochloride; His-AA represents histidine-histidine acetate; SA represents succinic acid-sodium succinate; 40℃ W4 represents storage at 40℃ for 4 weeks, the same applies below.

[0385] Example 2.2: pH Range Confirmation

[0386] Prepare 20 mM His-HCl buffer solutions with different pH values ​​as shown in Table 4, containing 150 mg / mL protein, 70 mg / mL sucrose, 0.6 mg / mL PS80, 0.1 mg / mL EDTA-2Na, 10 mM Met, and 2000 U / mL pH20. Conduct long-term stability studies on the samples (2–8 °C, i.e., 5 ± 3 °C for 6 months), using NR-CE and IEC as evaluation indicators to investigate the effect of different buffer pH values ​​on protein stability.

[0387] Long-term stability results (Table 4) show that after being stored at 2–8℃ for 6 months, the formulations with 20mM His-HCl at pH 5.0–5.8 all exhibited good stability. Among them, the formulations with the smallest changes in NR-CE fragments, IEC acid peak, and IEC alkaline peak were found under the condition of 20mM His-HCl at pH 5.3, indicating that the formulations were more stable.

[0388] Table 4. Results of pH Range Confirmation Experiment (2–8℃, M6)

[0389] Example 2.3: Surfactant Screening

[0390] Formulations were prepared using 20 mM His-HCl pH 5.3 buffer, 150 mg / mL protein, 70 mg / mL sucrose, and different concentrations of PS80 as shown in Table 5. Forced degradation studies were conducted on the samples (40°C for 4 weeks), and the effects of different surfactant concentrations on protein stability were investigated using SEC, NR-CE, and IEC as evaluation indicators.

[0391] The results are shown in Table 5. After being placed at 40℃ for 4 weeks, the differences in the SEC polymer peak, NR-CE fragments, and IEC acid peak results among the groups were small. Therefore, there were no significant differences among the formulation groups with PS80 concentrations of 0.1–0.6 mg / mL, and all formulations showed good stability.

[0392] Table 5. Surfactant screening results

[0393] Example 2.4: Effect of EDTA-2Na on formulation stability

[0394] Preparations were made using 20 mM His-HCl pH 5.3 buffer, 150 mg / mL protein, 70 mg / mL sucrose, 0.6 mg / mL PS80, and iron (Fe) ions as shown in Table 6, as well as formulations with or without EDTA-2Na and with 0.1 mg / mL EDTA-2Na. Forced degradation studies (40°C for 4 weeks) were conducted on the samples. The effects of EDTA-2Na on the stability of the formulations were investigated using appearance, SEC, NR-CE, IEC, and PS80 degradation rates as evaluation indicators.

[0395] The results are shown in Table 6. After 4 weeks of storage at 40℃, the formulation without EDTA-2Na turned yellow and PS80 was severely degraded. However, the formulation with EDTA-2Na remained clear after 4 weeks of storage at 40℃, with no significant degradation of PS80, and a significant reduction in the changes in SEC polymers, NR-CE fragments, IEC acid peak, and IEC basic peak. Therefore, EDTA-2Na not only prevents PS80 degradation but also maintains the stability of the formulation.

[0396] Table 6. Experimental results on the effect of EDTA-2Na on formulation stability Note: N / A means not added.

[0397] Example 2.5: Protein Concentration Screening

[0398] Formulations were prepared using 20 mM His-HCl pH 5.3 buffer, with different protein concentrations as shown in Table 7, 70 mg / mL sucrose, 0.6 mg / mL PS80, 0.1 mg / mL EDTA-2Na, 10 mM Met, and 2000 U / mL pH 20. Forced degradation studies (40°C for 4 weeks) were conducted on the samples, and the effects of different protein concentrations on the stability of the formulations were investigated using SEC, NR-CE, and IEC as evaluation indicators.

[0399] The results are shown in Table 7. After being placed at 40℃ for 4 weeks, the results showed that there were little difference between the groups, and the stability of the formulations with protein concentrations of 80–180 mg / mL was good.

[0400] Table 7. Protein concentration screening results

[0401] Example 2.6: Confirmation of sucrose concentration

[0402] A formulation was prepared containing 20 mM His-HCl pH 5.3 buffer, 150 mg / mL protein, 70 mg / mL sucrose, 0.6 mg / mL PS80, 0.1 mg / mL EDTA-2Na, 10 mM Met, and 2000 U / mL pH 20. The measured osmolarity was 306 mOsm / kg, close to the isotonic median, meeting the isotonic requirements of the formulation (270 mOsm / kg - 330 mOsm / kg).

[0403] Example 2.7: Prescription Confirmation

[0404] A formulation was prepared containing 20 mM His-HCl pH 5.3 buffer, 150 mg / mL protein, 70 mg / mL sucrose, 0.6 mg / mL PS80, 0.1 mg / mL EDTA-2Na, 10 mM Met, and 2000 U / mL pH 20. Long-term stability studies were conducted on the samples (stored at 2–8 °C for 6 months). The stability of the formulation was evaluated using SEC, NR-CE, IEC, and pH 20 enzyme activity residue as evaluation indicators.

[0405] The results are shown in Table 8. After being stored at 2–8℃ for 6 months, compared with D0, there were no significant changes in the SEC aggregate peak, NR-CE fragments, IEC acid peak, and PH20 enzyme activity, indicating that the protein in this formulation has good stability.

[0406] Table 8. Long-term stability results of the final prescription Note: 2~8℃ M6 means 6 months of storage at 2~8℃.

[0407] Although the invention has been described in detail with the aid of accompanying drawings and examples for clarity of understanding, these descriptions and examples should not be construed as limiting the scope of this disclosure. All patent and scientific literature disclosures cited herein are clearly and fully incorporated by reference.

Claims

1. A pharmaceutical composition comprising: Anti-PD-L1 antibody, and Hyaluronidase; The anti-PD-L1 antibody comprises a heavy chain variable region and a light chain variable region. The heavy chain variable region comprises HCDR1, HCDR2, and HCDR3, and the light chain variable region comprises LCDR1, LCDR2, and LCDR3. HCDR1 comprises the amino acid sequence of SEQ ID NO: 1, HCDR2 comprises the amino acid sequence of SEQ ID NO: 2, and HCDR3 comprises the amino acid sequence of SEQ ID NO:

3. LCDR1 comprises the amino acid sequence of SEQ ID NO: 4, LCDR2 comprises the amino acid sequence of SEQ ID NO: 5, and LCDR3 comprises the amino acid sequence of SEQ ID NO:

6.

2. The pharmaceutical composition according to claim 1, wherein the hyaluronidase is recombinant human hyaluronidase (rHuPH20) or a variant thereof; Preferably, the hyaluronidase comprises SEQ ID NO: 11, or an amino acid sequence having 80% sequence identity with it.

3. The pharmaceutical composition according to claim 1 or 2, wherein the hyaluronidase activity is 1000 U / mL to 3000 U / mL; Preferably, the hyaluronidase activity is 1600 U / mL to 2400 U / mL; More preferably, the hyaluronidase activity is 1800 U / mL to 2200 U / mL; Most preferably, the hyaluronidase has an enzyme activity of about 2000 U / mL.

4. The pharmaceutical composition according to any one of claims 1 to 3, wherein the concentration of the anti-PD-L1 antibody is from 80 mg / mL to 180 mg / mL; Preferably, the concentration of the anti-PD-L1 antibody is from 120 mg / mL to 180 mg / mL; More preferably, the concentration of the anti-PD-L1 antibody is from 135 mg / mL to 165 mg / mL; Most preferably, the concentration of the anti-PD-L1 antibody is about 150 mg / mL.

5. The pharmaceutical composition according to any one of claims 1 to 4, further comprising a buffer solution; Preferably, the buffer solution is histidine-histidine hydrochloride buffer, histidine-histidine acetate buffer, or succinate-sodium succinate buffer. More preferably, the buffer solution is a histidine-histidine hydrochloride buffer solution.

6. The pharmaceutical composition according to claim 5, wherein the concentration of the buffer solution is from 5 mM to 50 mM; Preferably, the concentration of the buffer solution is from 10 mM to 40 mM; More preferably, the concentration of the buffer solution is 15 mM to 30 mM; Most preferably, the concentration of the buffer solution is about 20 mM.

7. The pharmaceutical composition according to any one of claims 1 to 6, wherein the pH of the pharmaceutical composition is 5.0 to 5.8; Preferably, the pH of the pharmaceutical composition is 5.0 to 5.5; More preferably, the pH of the pharmaceutical composition is about 5.

3.

8. The pharmaceutical composition according to any one of claims 1 to 7, further comprising sugar; Preferably, the sugar is sucrose.

9. The pharmaceutical composition according to claim 8, wherein the concentration of the sugar is from 50 mg / mL to 90 mg / mL; Preferably, the concentration of the sugar is from 55 mg / mL to 85 mg / mL; Preferably, the concentration of the sugar is 65 mg / mL to 75 mg / mL; More preferably, the concentration of the sugar is about 70 mg / mL.

10. The pharmaceutical composition according to any one of claims 1 to 9, further comprising polysorbate; Preferably, the polysorbate is polysorbate 80 or polysorbate 20; More preferably, the polysorbate is polysorbate 80.

11. The pharmaceutical composition according to claim 10, wherein the concentration of the polysorbate is from 0.1 mg / mL to 1.0 mg / mL; Preferably, the concentration of the polysorbate is from 0.1 mg / mL to 0.8 mg / mL; More preferably, the concentration of the polysorbate is from 0.4 mg / mL to 0.8 mg / mL; Most preferably, the concentration of the polysorbate is about 0.6 mg / mL.

12. The pharmaceutical composition according to any one of claims 1 to 11, further comprising a metal ion chelating agent; Preferably, the metal ion chelating agent is disodium edetate.

13. The pharmaceutical composition according to claim 12, wherein the concentration of the metal ion chelating agent is from 0.01 mg / mL to 1.0 mg / mL; Preferably, the concentration of the metal ion chelating agent is from 0.01 mg / mL to 0.5 mg / mL; More preferably, the concentration of the metal ion chelating agent is from 0.05 mg / mL to 0.2 mg / mL; Most preferably, the concentration of the metal ion chelating agent is about 0.1 mg / mL.

14. The pharmaceutical composition according to any one of claims 1 to 13, further comprising methionine.

15. The pharmaceutical composition according to claim 14, wherein the concentration of methionine is from 1 mM to 20 mM; Preferably, the concentration of the methionine is 5 mM to 15 mM; More preferably, the concentration of the methionine is about 10 mM.

16. The pharmaceutical composition according to any one of claims 1 to 15, wherein the anti-PD-L1 antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 7, or an amino acid sequence having at least 80% sequence identity therewith; and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 8, or an amino acid sequence having at least 80% sequence identity therewith; Preferably, the anti-PD-L1 antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 7, and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 8; More preferably, the anti-PD-L1 antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 9, or an amino acid sequence having at least 80% sequence identity with it; and the light chain comprises the amino acid sequence of SEQ ID NO: 10, or an amino acid sequence having at least 80% sequence identity with it. Most preferably, the anti-PD-L1 antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 9, and the light chain comprises the amino acid sequence of SEQ ID NO:

10.

17. The pharmaceutical composition according to any one of claims 1 to 16, comprising the following components: a) Anti-PD-L1 antibody at concentrations of 80 mg / mL to 180 mg / mL; b) 5mM to 50mM histidine-histidine hydrochloride buffer; c) Sucrose at concentrations of 50 mg / mL to 90 mg / mL; d) Polysorbate 80 at concentrations of 0.1 mg / mL to 1.0 mg / mL; e) Disodium edetate at concentrations of 0.01 mg / mL to 1.0 mg / mL; f) Methionine from 1 mM to 20 mM; 1000 U / mL to 3000 U / mL of hyaluronidase; Furthermore, the pH of the pharmaceutical composition is 5.0 to 5.8; Preferably, the pharmaceutical composition comprises the following components: a) Anti-PD-L1 antibody at concentrations of 120 mg / mL to 180 mg / mL; b) 10 mM to 40 mM histidine-histidine hydrochloride buffer; c) Sucrose at concentrations of 55 mg / mL to 85 mg / mL; d) Polysorbate 80 at concentrations of 0.1 mg / mL to 0.8 mg / mL; e) Disodium edetate at concentrations of 0.01 mg / mL to 0.5 mg / mL; f) Methionine from 5 mM to 15 mM; 1600 U / mL to 2400 U / mL of hyaluronidase; Furthermore, the pH of the pharmaceutical composition is 5.0 to 5.8; More preferably, the pharmaceutical composition comprises the following components: a) Anti-PD-L1 antibody at concentrations of 135 mg / mL to 165 mg / mL; b) 15mM to 30mM histidine-histidine hydrochloride buffer; c) Sucrose at concentrations of 65 mg / mL to 75 mg / mL; d) Polysorbate 80 at concentrations of 0.4 mg / mL to 0.8 mg / mL; e) Disodium edetate at concentrations of 0.05 mg / mL to 0.2 mg / mL; f) Methionine from 5 mM to 15 mM; 1800 U / mL to 2200 U / mL of hyaluronidase; Furthermore, the pH of the pharmaceutical composition is 5.0 to 5.5; Most preferably, the pharmaceutical composition comprises the following components: a) Approximately 150 mg / mL of anti-PD-L1 antibody; b) Approximately 20 mM histidine-histidine hydrochloride buffer; c) Approximately 70 mg / mL of sucrose; d) Approximately 0.6 mg / mL of polysorbate 80; e) Approximately 0.1 mg / mL of disodium edetate; f) Approximately 10 mM of methionine; g) Approximately 2000 U / mL of hyaluronidase; The pH of the pharmaceutical composition is approximately 5.

3.

18. The pharmaceutical composition according to any one of claims 1 to 17, wherein it is a subcutaneous injection formulation.

19. An article comprising a container containing a pharmaceutical composition as described in any one of claims 1 to 18.

20. Use of the pharmaceutical composition according to any one of claims 1 to 18 in the preparation of a medicament for treating or preventing a disease; Preferably, the disease is a tumor; More preferably, the disease is breast cancer, lung cancer, stomach cancer, intestinal cancer, kidney cancer, melanoma, liver cancer, esophageal cancer, cervical cancer, ovarian cancer, endometrial cancer, biliary tract cancer, pancreatic cancer, thyroid cancer, bladder cancer, nasopharyngeal cancer, prostate cancer, or squamous cell carcinoma.

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