Neoadjuvant immunotherapy using PD-L1 inhibitors in treatment of cancer

Abstract

The present disclosure relates to methods for treating a tumor or inhibiting tumor proliferation comprising administering to a cancer patient in need thereof a therapeutically effective amount of a programmed death ligand 1 (PD-L1) inhibitor (e.g., an anti-PD-L1 antibody) as a neoadjuvant therapy followed by surgical resection. In particular, the present disclosure relates to methods for administering a PD-L1 inhibitor as a pre-operative therapy to a patient suffering from resectable upper gastrointestinal cancer (e.g., resectable localized gastric, esophageal or liver cancer). Administration of a PD-L1 inhibitor, in particular an anti-PD-L1 antibody, has proved to exhibit an excellent therapeutic effect as a pre-operative therapy for patients suffering from resectable localized gastric, esophageal and liver cancer.

Description

Technical Field

[0001] This disclosure relates to methods for treating tumors and inhibiting tumor proliferation, the methods comprising administering a therapeutically effective amount of a programmed death-ligand 1 (PD-L1) inhibitor (e.g., anti-PD-L1 antibody IMC-001 or its bioequivalence) to a cancer patient as neoadjuvant therapy, followed by surgical resection. In particular, this disclosure relates to methods for administering a PD-L1 inhibitor as preoperative therapy to patients with surgically resectable upper gastrointestinal cancers (e.g., localized gastric cancer, esophageal cancer, or liver cancer).

[0002] sequence list

[0003] This application contains a sequence list that has been electronically submitted in XML format, which is incorporated herein by reference in its entirety. A copy of the sequence list was created on April 13, 2024, and is named KC24060.xml, with a size of 12.3 kilobytes. Background Technology

[0004] Immune evasion, as one of the major characteristics of cancer, has recently gained attention, and advances in tumor immunology have begun to elucidate its underlying mechanisms. Immune checkpoints are a concept encompassing immunosuppressive regulatory molecules that prevent immune cells from attacking their own cells. Tumor cells utilize these immune checkpoints as one of several mechanisms to evade immune surveillance, thereby creating an immunosuppressive environment within the tumor.

[0005] The therapeutic strategy of cancer immunotherapy, which exerts its anti-cancer effect by activating the suppressed anti-tumor immune system, has attracted considerable attention. Representative immune checkpoint inhibitors (such as CTLA-4 inhibitors or PD-1 / PD-L1 inhibitors) have recently been shown to improve survival rates in various cancers. Therefore, cancer immunotherapy has become one of the standard cancer treatments and has pioneered a new paradigm in cancer treatment (Pardoll DM: The blockade of immune checkpoints in cancer immunotherapy. Nat Rev Cancer 12:252-64, 2012).

[0006] Research on lung cancer has shown that immune checkpoint inhibitors exhibit stronger therapeutic effects in tumors with a higher number of mutations, especially in cases of high clonal neoantigen burden. However, even in tumors with a high total number of mutations, PD-1 inhibitors show reduced therapeutic effects when the subclonal neoantigen burden is high (McGranahan N, Furness AJ, Rosenthal R et al.: Clonal neoantigens elicit T cell immunoreactivity and sensitivity to immune checkpoint blockade. Science 351:1463-9, 2016).

[0007] Furthermore, considering the increase in the number of subclonal neoantigens and the progression of the immunosuppressive microenvironment as tumors evolve with cancer progression, immune checkpoint inhibitors may be more effective in treating early-stage localized cancers than in stage IV metastatic cancers (Jamal-Hanjani M, Quezada SA, Larkin J et al.: Translational implications of tumorheterogeneity. Clin Cancer Res 21:1258-66, 2015; and Gil Del Alcazar CR, Huh SJ, Ekram MB et al.: Immune Escape in Breast Cancer During In Situ to Invasive Carcinoma Transition. Cancer Discov 7:1098-1115, 2017).

[0008] Immune checkpoint inhibitors (such as pembrolizumab, nivolumab, and ipilimumab) have been shown to improve survival or shrink tumor size in patients with unresectable locally advanced or metastatic gastric, esophageal, and hepatocellular carcinomas, thus demonstrating the effectiveness of treatment strategies using immune checkpoint inhibitors for these cancers.

[0009] On the other hand, radical surgical resection is a treatment option for resectable gastric, esophageal, and hepatocellular carcinomas. However, tumor recurrence after surgery is common, and early recurrence has also been observed. Therefore, there is an urgent need for safe and effective therapies to treat surgically resectable cancers. Summary of the Invention Technical issues

[0010] The purpose of this disclosure is to solve all the aforementioned problems in the prior art.

[0011] The purpose of this disclosure is to provide preoperative therapies for patients with surgically resectable upper gastrointestinal cancers, methods for treating cancer, pharmaceutical compositions, kits, and dosing regimens for treating cancer.

[0012] The purpose of this disclosure is not limited to the foregoing. The purpose of this disclosure will become clearer from the following description and can be achieved by the means set forth in the claims and combinations thereof. Solution to the problem

[0013] The following are representative ways in which this disclosure achieves the above objectives.

[0014] According to an aspect of this disclosure, a pharmaceutical composition is provided for treating or inhibiting tumor proliferation in a patient with upper gastrointestinal cancer, said composition comprising a therapeutically effective amount of a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody that specifically binds to PD-L1, comprising HCDR1, HCDR2, and HCDR3 contained in the heavy chain variable region (HCVR) of SEQ ID NO:1, and LCDR1, LCDR2, and LCDR3 contained in the light chain variable region (LCVR) of SEQ ID NO:2), wherein said composition is administered to said patient as neoadjuvant therapy prior to surgical resection of the tumor.

[0015] According to another aspect of this disclosure, a method for treating a tumor or inhibiting tumor growth is provided, the method comprising: (a) selecting a patient with upper gastrointestinal cancer; (b) administering to the patient a therapeutically effective amount of a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody that specifically binds to PD-L1, comprising HCDR1, HCDR2, and HCDR3 contained in the heavy chain variable region (HCVR) of SEQ ID NO:1, and LCDR1, LCDR2, and LCDR3 contained in the light chain variable region (LCVR) of SEQ ID NO:2); and (c) surgically removing the tumor of the upper gastrointestinal cancer after step (b).

[0016] In this implementation, upper gastrointestinal cancers may include stomach cancer, esophageal cancer, or liver cancer.

[0017] In an embodiment, the anti-PD-L1 antibody may include a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region includes HCDR1 of SEQ ID NO:5, HCDR2 of SEQ ID NO:6 and HCDR3 of SEQ ID NO:7, and the light chain variable region includes LCDR1 of SEQ ID NO:8, LCDR2 of SEQ ID NO:9 and LCDR3 of SEQ ID NO:10.

[0018] In an embodiment, the light chain variable region of the anti-PD-L1 antibody may contain the amino acid sequence of SEQ ID NO:2 or an amino acid sequence that is at least 95% identical to SEQ ID NO:2, or may be composed of such sequence.

[0019] In an embodiment, the heavy chain variable region of the anti-PD-L1 antibody may contain the amino acid sequence of SEQ ID NO:1 or an amino acid sequence that is at least 95% identical to SEQ ID NO:1, or may be composed of such sequence.

[0020] In an embodiment, the anti-PD-L1 antibody may comprise a heavy chain having the amino acid sequence SEQ ID NO:3 and a light chain having the amino acid sequence SEQ ID NO:4.

[0021] In one embodiment, the anti-PD-L1 antibody may be a fully human antibody.

[0022] In this embodiment, the anti-PD-L1 antibody can retain the Fc effector function that can stimulate antibody-dependent cytotoxicity (ADCC).

[0023] In this embodiment, the anti-PD-L1 antibody may be a fully human antibody of the IgG class.

[0024] In this embodiment, the anti-PD-L1 antibody may be a fully human antibody of the IgG1 or IgG4 class.

[0025] In one implementation, the neoadjuvant therapy allows the anti-PD-L1 antibody to be administered intravenously or subcutaneously.

[0026] In this implementation, the upper gastrointestinal cancer may be resectable.

[0027] In this implementation, the upper gastrointestinal cancer may be recurrent.

[0028] In this implementation, the upper gastrointestinal cancer may be metastatic.

[0029] In this embodiment, the upper gastrointestinal cancer can be treated with radical surgery.

[0030] In this embodiment, the gastric cancer may be a submucosal tumor of the stomach or adenocarcinoma of the stomach (GC).

[0031] In this embodiment, the esophageal cancer may be esophageal squamous cell carcinoma (ESCC).

[0032] In this embodiment, the liver cancer may be hepatocellular carcinoma (HCC).

[0033] In one embodiment, the neoadjuvant therapy allows the anti-PD-L1 antibody to be administered once or multiple times, with each dose administered at intervals of 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, or 6 weeks.

[0034] In one implementation, the neoadjuvant therapy allows the anti-PD-L1 antibody to be administered once or multiple times, with each dose administered at 2-week intervals.

[0035] In one implementation, the neoadjuvant therapy allows the anti-PD-L1 antibody to be administered once, twice, three times, four times, five times, or six times prior to surgical removal of the tumor.

[0036] In one implementation, the neoadjuvant therapy allows the anti-PD-L1 antibody to be administered at a dose of 10 mg / kg to 30 mg / kg based on the patient's weight.

[0037] In one embodiment, the neoadjuvant therapy allows the anti-PD-L1 antibody to be administered at a dose of 20 mg / kg.

[0038] In one embodiment, surgical resection of the tumor may be performed 11 to 84 days or 11 to 42 days after the last administration of the anti-PD-L1 antibody.

[0039] According to another aspect of this disclosure, a kit is provided comprising a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody that specifically binds to PD-L1, comprising HCDR1, HCDR2, and HCDR3 contained in the heavy chain variable region (HCVR) of SEQ ID NO:1, and LCDR1, LCDR2, and LCDR3 contained in the light chain variable region (LCVR) of SEQ ID NO:2) as a neoadjuvant therapy for treating tumors or inhibiting tumor proliferation in patients with upper gastrointestinal cancer, and a manual for using said PD-L1 inhibitor.

[0040] In one embodiment, the manual included in the kit includes administering a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody that specifically binds to PD-L1, comprising HCDR1, HCDR2, and HCDR3 contained in the heavy chain variable region (HCVR) of SEQ ID NO:1, and LCDR1, LCDR2, and LCDR3 contained in the light chain variable region (LCVR) of SEQ ID NO:2) once, twice, three times, four times, five times, or six times as neoadjuvant therapy to patients with upper gastrointestinal cancer at intervals of 10 to 18 days prior to surgical resection of the tumor.

[0041] According to yet another aspect of this disclosure, the use of a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody that specifically binds to PD-L1, comprising HCDR1, HCDR2, and HCDR3 contained in the heavy chain variable region (HCVR) of SEQ ID NO:1, and LCDR1, LCDR2, and LCDR3 contained in the light chain variable region (LCVR) of SEQ ID NO:2) in a method for treating a tumor or inhibiting tumor growth, the method comprising: (a) selecting a patient with upper gastrointestinal cancer; (b) administering a therapeutically effective amount of the PD-L1 inhibitor to the patient; and (c) surgically removing the tumor of the upper gastrointestinal cancer after step (b).

[0042] According to yet another aspect of this disclosure, a method for treating a tumor or inhibiting tumor growth is provided, the method comprising: (a) selecting a patient with resectable gastric cancer; (b) administering to the patient a therapeutically effective amount of a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody that specifically binds to PD-L1, comprising HCDR1, HCDR2, and HCDR3 contained in the heavy chain variable region (HCVR) of SEQ ID NO:1, and LCDR1, LCDR2, and LCDR3 contained in the light chain variable region (LCVR) of SEQ ID NO:2); and (c) surgically removing the gastric cancer tumor after step (b).

[0043] According to yet another aspect of this disclosure, a method for treating a tumor or inhibiting tumor growth is provided, the method comprising: (a) selecting a patient with resectable esophageal cancer; (b) administering to the patient a therapeutically effective amount of a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody that specifically binds to PD-L1, comprising HCDR1, HCDR2, and HCDR3 contained in the heavy chain variable region (HCVR) of SEQ ID NO:1, and LCDR1, LCDR2, and LCDR3 contained in the light chain variable region (LCVR) of SEQ ID NO:2); and (c) surgically removing the esophageal cancer tumor after step (b).

[0044] According to yet another aspect of this disclosure, a method for treating a tumor or inhibiting tumor growth is provided, the method comprising: (a) selecting a patient with resectable hepatocellular carcinoma; (b) administering to the patient a therapeutically effective amount of a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody that specifically binds to PD-L1, comprising HCDR1, HCDR2, and HCDR3 contained in the heavy chain variable region (HCVR) of SEQ ID NO:1, and LCDR1, LCDR2, and LCDR3 contained in the light chain variable region (LCVR) of SEQ ID NO:2); and (c) surgically removing the hepatocellular carcinoma tumor after step (b). Invention Effects

[0045] Neoadjuvant therapy using anti-PD-L1 antibodies has demonstrated good tolerability and promising antitumor activity in patients with resectable microsatellite-stable gastric, esophageal, and hepatocellular carcinoma. In the examples disclosed herein, administration of IMC-001 as preoperative therapy resulted in low-grade treatment-related adverse events (TRAEs); partial response (PR) or stable disease (SD) in patients with measurable lesions; and metabolic partial response (mPR), stable metabolic disease (mSD), or metabolic disease progression (mPD) in patients with evaluable metabolic responses. Furthermore, it has been determined that administration of IMC-001 as preoperative therapy leads to varying degrees of tumor necrosis or fibrosis and results in a reduction of at least 50% of residual viable tumor cells in some patients. Attached Figure Description

[0046] Figure 1 A clinical trial process according to an embodiment of this disclosure is illustrated schematically.

[0047] Figure 2 Clinical oncology responses as clinical trial results according to embodiments of this disclosure are shown. PR indicates partial response, and SD indicates stable disease.

[0048] Figure 3 Clinical oncology responses as clinical trial results according to embodiments of this disclosure are shown. mPR indicates a metabolic partial response, mSD indicates stable metabolic disease, and mPD indicates metabolic disease progression.

[0049] Figure 4 The pathological response as a clinical trial result is shown according to an embodiment of the present disclosure. Detailed Implementation

[0050] The detailed description of this disclosure described below will be illustrated with reference to specific accompanying drawings (if any), which relate to particular embodiments in which this disclosure may be practiced. However, this disclosure is not limited thereto, but is limited only by the appended claims and covers any scope identical or equivalent to that stated in the claims. It should be understood that the various embodiments / examples of this disclosure are different from each other, but are not necessarily mutually exclusive. For example, certain shapes, structures, and features described herein may be varied in one embodiment / example to another, or may be implemented by a combination of multiple embodiments / examples, without departing from the technical concept and scope of this disclosure. Unless otherwise defined, the technical and academic terms used in this specification have the same meaning as those commonly used in the art to which this disclosure pertains. In case of conflict, the specification containing the definition shall prevail. The definitions of terms used in this specification shall be used to interpret the purposes of this specification, and terms expressed in the singular shall be interpreted as also referring to the plural form (i.e., at least one), unless the context makes it inappropriate, and vice versa.

[0051] Methods used to treat cancer or inhibit cancer proliferation

[0052] In this disclosure, methods for treating tumors or inhibiting tumor proliferation are provided, the methods comprising selecting a patient with surgically resectable upper gastrointestinal cancer (e.g., localized gastric cancer, esophageal cancer, or liver cancer) and administering a PD-L1 inhibitor (e.g., anti-PD-L1 antibody IMC-001 or its bioequivalent) to the patient as neoadjuvant therapy prior to surgical resection of the patient's tumor.

[0053] In implementation, the methods disclosed herein may also include administering a PD-L1 inhibitor (e.g., anti-PD-L1 antibody IMC-001 or its bioequivalence) to a patient as adjuvant therapy after surgery for the treatment of upper gastrointestinal cancers (e.g., gastric cancer, esophageal cancer, or liver cancer).

[0054] As used herein, "gastric cancer" refers to any cancer (malignant tumor) occurring in the stomach, including gastric adenocarcinoma, lymphoma, submucosal tumors of the stomach, and leiomyosarcoma. Gastric cancer can occur in any part of the stomach and can spread throughout the stomach and to other organs, particularly the esophagus, lungs, and liver. In this embodiment, the gastric cancer is gastric adenocarcinoma. In this embodiment, the gastric cancer is resectable and recurrent. In this embodiment, the gastric cancer is metastatic.

[0055] As used herein, "esophageal cancer" refers to cancer occurring in the esophagus. Based on its location, it is classified into cervical esophageal cancer, thoracic esophageal cancer, and gastroesophageal junction cancer. Based on cell morphology, it is classified into squamous cell carcinoma, adenocarcinoma, sarcoma, lymphoma, melanoma, etc. In this embodiment, the esophageal cancer is esophageal squamous cell carcinoma (ESCC). In this embodiment, the esophageal cancer is resectable and recurrent. In this embodiment, the esophageal cancer is metastatic.

[0056] As used herein, "liver cancer" refers to cancer of the liver, such as hepatocellular carcinoma, fibrolamellar carcinoma, cholangiocarcinoma, angiosarcoma, and hepatoblastoma. In this embodiment, the liver cancer is hepatocellular carcinoma (HCC). In this embodiment, the liver cancer is resectable and recurrent. In this embodiment, the liver cancer is metastatic.

[0057] In some embodiments, the patient is a surgical candidate for resection of a tumor of the stomach, esophagus, or liver. In some embodiments, the patient has stomach, esophagus, or liver cancer and is expected to undergo radical surgery.

[0058] As used herein, the term "treatment / management" and the like mean relieving or reducing the severity of at least one symptom or indication; temporarily or permanently eliminating the triggers of symptoms; slowing or inhibiting tumor growth; reducing tumor cell burden or tumor load; promoting tumor regression; shrinking, necrosis, and / or disappearing of tumors; preventing tumor recurrence; preventing or inhibiting metastasis; inhibiting the growth of metastatic tumors; eliminating the need for surgery; and / or increasing the survival of the subject. In many implementations, the terms "tumor," "lesion," "tumor lesion," "cancer," and "malignant tumor" are used interchangeably to refer to one or more cancerous growths.

[0059] As used herein, the term "recurrence" refers to the frequent or repeated diagnoses of gastric, esophageal, or liver cancer in a patient, or the frequent or repeated appearance of a single tumor (e.g., a primary tumor and / or a new tumor that may represent a recurrence of a previous tumor). In some implementations, administration of a PD-L1 inhibitor suppresses recurrence of gastric, esophageal, or liver cancer tumors in the patient.

[0060] As used herein, the expression "subject requiring a PD-L1 inhibitor" refers to a human or non-human mammal exhibiting one or more symptoms or indications of gastric, esophageal, or hepatocellular carcinoma, and / or who has been diagnosed with gastric, esophageal, or hepatocellular carcinoma and requires treatment. In many embodiments, the terms "subject" and "patient" are used interchangeably. This expression includes subjects with a primary, confirmed, or recurrent tumor (advanced malignancy). In some embodiments, this expression includes subjects with and / or requiring treatment for recurrent but non-metastatic gastric, esophageal, or hepatocellular carcinoma. In some embodiments, this expression includes patients with solid tumors that are resistant, refractory, or poorly controlled to prior treatment (e.g., surgery or treatment with other anticancer drugs besides IMC-001 or its bioequivalent). In some embodiments, this expression includes subjects with gastric, esophageal, or hepatocellular carcinoma who are not candidates for radical surgery.

[0061] In some embodiments, the methods of this disclosure are used to treat a subject with a solid tumor. As used herein, the term "solid tumor" refers to an abnormal mass of tissue that does not typically contain cysts or fluid-filled areas. Solid tumors can be benign (non-cancerous) or malignant (cancerous). For the purposes of this disclosure, the term "solid tumor" refers to a malignant solid tumor. The term includes different types of solid tumors named after the cell types that form the tumor, namely sarcomas, carcinomas, and blastomas.

[0062] In some embodiments, the disclosed method includes administering a therapeutically effective amount of a PD-L1 inhibitor (e.g., anti-PD-L1 antibody IMC-001 or its bioequivalence) in combination with additional therapeutic agents or therapies. These additional therapeutic agents or therapies may be administered to increase antitumor efficacy, reduce the toxicity of one or more therapies, and / or reduce the dosage of one or more therapies. In various embodiments, the additional therapeutic agents or therapies may include one or more of the following: antiviral therapy (e.g., cidofovir), photodynamic therapy, PD-1 inhibitors (e.g., antibodies against PD-1 known in the art, such as nivolumab or pembrolizumab), lymphocyte activation gene 3 (LAG3) inhibitors (e.g., anti-LAG3 antibodies), cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) inhibitors (e.g., ipilimumab), and glucocorticoid-induced tumor necrosis factor receptor (GITR) agonists (e.g., anti-GITR antibodies). T cell immunoglobulin and mucin-3 (TIM3) inhibitors, B and T lymphocyte attenuator (BTLA) inhibitors, T cell immune receptor (TIGIT) inhibitors with Ig and ITIM domains, CD38 inhibitors, CD47 inhibitors, other T cell co-inhibitors or ligand antagonists (e.g., antibodies against CD-28, 2B4, LY108, LAIR1, ICOS, CD160, or VISTA), CD20 inhibitors (e.g., anti-CD20 antibodies or bispecific CD3 / CD20 antibodies), indoleamine-2,3-dioxygenase (IDO) inhibitors, CD28 activators, vascular endothelial growth factor (VEGF) antagonists (e.g., “VEGF-Trap”, such as aflibercept or other VEGF inhibitory fusion proteins (as listed in US7087411), or anti-VEGF antibodies or their antigen-binding fragments (e.g., bevacizumab or ranibizumab), or small molecule kinase inhibitors of the VEGF receptor (e.g., sunitinib, sorafenib, pazopanib, or ramucirumab), angiopoietin-2 (Ang2) inhibitors, transforming growth factor β (TGFβ) inhibitors, epidermal growth factor receptor (EGFR) inhibitors (e.g., erlotinib, cetuximab), co-stimulatory receptor agonists (e.g., CD28, 4-1BB, or OX40 agonists), tumor-specific antigens (e.g., CA9, CA125, melanoma-associated antigen 3 (MAGE3), carcinoembryonic antigen (CEA), vimentin, tumor-M2-PK, prostate-specific antigen (PSA) A) Antibodies against mucin-1, MART-1, and CA19-9; vaccines (e.g., BCG or cancer vaccines); adjuvants that enhance antigen presentation (e.g., granulocyte-macrophage colony-stimulating factor); oncolytic viruses; cytotoxins; chemotherapeutic agents (e.g., pemetrexed, dacarbazine, temozolomide, cyclophosphamide, docetaxel, doxorubicin, daunorubicin, cisplatin, carboplatin, gemcitabine, methotrexate, mitoxantrone, oxaliplatin, paclitaxel, topotecan, irinotecan, vinorelbine, and... Vincristine, platinum-based chemotherapy (e.g., platinum-doublet chemotherapy), tyrosine kinase inhibitors (e.g., lenvatinib, regorafenib, and cabozantinib), IL-6R inhibitors, IL-4R inhibitors, IL-10 inhibitors, cytokines (e.g., IL-2, IL-7, IL-12, IL-21, and IL-15), antibody-drug conjugates (ADCs) (e.g., anti-CD19-DM4 ADCs and anti-DS6-DM4 ADCs), chimeric antigen receptor T cells (e.g., CD19-targeting T cells), anti-inflammatory drugs (e.g., corticosteroids), nonsteroidal anti-inflammatory drugs (NSAIDs), and dietary supplements (e.g., antioxidants).

[0063] As used herein, the term "antiviral therapy" refers to any agent, medicine, or therapy used to treat, prevent, or improve viral infection in a host subject, including but not limited to: zidovudine, lamivudine, abacavir, ribavirin, lopinavir, efavirenz, cobicistat, tenofovir, rilpivirine, analgesics, corticosteroids, and combinations thereof. In the context of this disclosure, chronic viral infection includes infections caused by viruses, including but not limited to human immunodeficiency virus (HIV), hepatitis B virus (HBV), and hepatitis C virus (HCV).

[0064] In some implementations, administering a therapeutically effective amount of a PD-L1 inhibitor (e.g., anti-PD-L1 antibody IMC-001 or its bioequivalence) to a subject with gastric, esophageal, or liver cancer results in enhanced tumor proliferation inhibition, for example, higher tumor regression in the treated subject.

[0065] In some implementations, administration of a PD-L1 inhibitor results in one or more of the following: (i) an improved tumor response rate, such as an improved overall response rate, complete response, or partial response, compared to subjects treated with surgical resection alone or untreated subjects; (ii) delayed tumor growth and progression (e.g., in treated subjects, tumor growth may be delayed by about 3 days, more than 3 days, about 7 days, more than 7 days, more than 15 days, more than 1 month, more than 3 months, more than 6 months, more than 1 year, more than 2 years, or more than 3 years, compared to subjects treated with surgical resection alone or untreated subjects); (iii) an increased disease-free survival (DFS) from the date of treatment until tumor recurrence or death, compared to subjects treated with surgical resection alone or untreated subjects; and (iv) an improved overall response rate, complete response, or partial response, compared to subjects treated with surgical resection alone or untreated subjects. In some implementations, administering a therapeutically effective amount of a PD-L1 inhibitor (e.g., anti-PD-L1 antibody IMC-001 or its bioequivalence) to a subject with gastric, esophageal, or hepatic cancer may prevent tumor recurrence and / or increase the subject's survival time, for example, by more than 15 days, more than 1 month, more than 3 months, more than 6 months, more than 12 months, more than 18 months, more than 24 months, more than 36 months, or more than 48 months compared to a subject treated with surgical resection alone or a treatment-naïve subject.

[0066] In some embodiments, administration of a therapeutically effective amount of a PD-L1 inhibitor (e.g., anti-PD-L1 antibody IMC-001 or its bioequivalence) to a subject with gastric, esophageal, or hepatocellular carcinoma results in an increase in overall survival (OS) or progression-free survival (PFS) compared to subjects treated with surgical resection alone. In some embodiments, the PFS increase is at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 1 year, at least 2 years, or at least 3 years compared to subjects treated with surgical resection alone. In some embodiments, the OS increase is at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 1 year, at least 2 years, or at least 3 years compared to subjects treated with surgical resection alone.

[0067] PD-L1 inhibitors

[0068] The methods disclosed herein include administering a therapeutically effective amount of a PD-L1 inhibitor, wherein the PD-L1 inhibitor is an anti-PD-L1 antibody, such as the fully human anti-PD-L1 antibody IMC-001 or its bioequivalence. As used herein, the term "bioequivalence" refers to an anti-PD-L1 antibody or PD-L1 binding protein or fragment thereof, which is a pharmaceutical equivalent or substitute, and whose absorption rate and / or extent, when administered at the same molar dose (single or multiple doses) under similar experimental conditions, does not show a significant difference from that of the IMC-001 antibody. In the context of this disclosure, the term "bioequivalence" includes an antigen-binding protein that binds to PD-L1 and is not clinically different from the IMC-001 antibody in terms of safety, purity, and / or potency.

[0069] As used herein, the term "antibody" is intended to refer to an immunoglobulin molecule (i.e., a "complete antibody molecule") consisting of four polypeptide chains (two heavy chains (H) and two light chains (L) linked together by disulfide bonds), as well as its multimer (e.g., IgM) or its antigen-binding fragment. Each heavy chain consists of a heavy chain variable region ("HCVR" or "VH") and a heavy chain constant region (consisting of domains CH1, CH2, and CH3). Each light chain consists of a light chain variable region ("LCVR" or "VL") and a light chain constant region (CL). The VH and VL regions can be further subdivided into hypervariable regions (called complementarity-determining regions (CDRs)) dispersed with more conserved regions (called framework regions (FRs)). Each VH and VL consists of three CDRs and four FRs, arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In some embodiments, the FR of the antibody (or its antigen-binding fragment) may be identical to the human germline sequence, or may be natural or artificially modified. The common amino acid sequence may be defined based on a side-by-side analysis of two or more CDRs. As used herein, the term "antibody" also includes the antigen-binding fragment of the complete antibody molecule.

[0070] As used herein, the terms “antigen-binding fragment” and “antigen-binding portion” of an antibody include any naturally occurring, enzymatically obtainable, synthetic, or genetically engineered polypeptide or glycoprotein that specifically binds to an antigen to form a complex. Antigen-binding fragments of antibodies can be obtained from intact antibody molecules, for example, using any suitable standard technique, such as proteolytic digestion or recombinant genetic engineering techniques involving the manipulation and expression of DNA encoding variable and optional constant domains of the antibody. Such DNA is known and / or readily available from, for example, commercial sources, DNA libraries (including, for example, phage antibody libraries), or can be synthesized. DNA can be sequenced and manipulated by chemical methods or by using molecular biology techniques, such as arranging one or more variable and / or constant domains into suitable conformations, or introducing codons, generating cysteine ​​residues, modifying, adding, or deleting amino acids, etc.

[0071] Non-limiting examples of antigen-binding fragments include: (i) Fab fragments; (ii) F(ab')2 fragments; (iii) Fd fragments; (iv) Fv fragments; (v) single-chain Fv (scFv) molecules; (vi) dAb fragments; and (vii) minimal recognition units consisting of amino acid residues mimicking the hypervariable region of an antibody (e.g., isolated complementarity-determining regions (CDRs), such as CDR3 peptides), or constrained FR3-CDR3-FR4 peptides. The term "antigen-binding fragment" as used herein also encompasses other engineered molecules, such as domain-specific antibodies, single-domain antibodies, domain-deficient antibodies, chimeric antibodies, CDR-transplanted antibodies, diabodies, triabodies, tetrabodies, minibodies, nanobodies (e.g., monovalent nanobodies, bivalent nanobodies, etc.), small modular immunopharmaceuticals (SMIPs), and shark variable IgNAR domains.

[0072] Antigen-binding fragments of antibodies will typically contain at least one variable domain. The variable domain can be of any size or amino acid composition and will typically contain at least one CDR adjacent to or in frame one or more frames. In antigen-binding fragments having a VH domain associated with a VL domain, the VH and VL domains can be positioned relative to each other in any suitable manner. For example, the variable region can be a dimer and may include VH-VH, VH-VL, or VL-VL dimers. Alternatively, the antigen-binding fragment of an antibody may contain a monomeric VH or VL domain.

[0073] In some embodiments, the antigen-binding fragment of the antibody may include at least one variable domain covalently linked to at least one constant domain. Non-limiting exemplary configurations of the variable and constant domains that may be present in the antigen-binding fragment of the antibody disclosed herein include: (i) VH-CH1; (ii) VH-CH2; (iii) VH-CH3; (iv) VH-CH1-CH2; (v) VH-CH1-CH2-CH3; (vi) VH-CH2-CH3; (vii) VH-CL; (viii) VL-CH1; (ix) VL-CH2; (x) VL-CH3; (xi) VL-CH1-CH2; (xii) VL-CH1-CH2-CH3; (xiii) VL-CH2-CH3; and (xiv) VL-CL. In any configuration of the variable and constant domains, including any of the exemplary configurations listed above, the variable and constant domains may be directly interconnected or connected via complete or partial hinge or linker regions. The hinge region may consist of at least two (e.g., 5, 10, 15, 20, 40, 60, or more) amino acids, resulting in flexible or semi-flexible connections between adjacent variable and / or constant domains in a single polypeptide molecule. Furthermore, the antigen-binding fragment of the antibody disclosed herein may comprise homodimers or heterodimers (or other multimers) of any of the variable and constant domain configurations listed above, which are connected to each other and / or to one or more monomeric VH or VL domains via non-covalent bonds (e.g., via disulfide bonds).

[0074] The antibodies used in the methods disclosed herein may be human antibodies. As used herein, the term "human antibody" refers to an antibody having variable and constant regions derived from human germline immunoglobulin sequences. However, the human antibodies of this disclosure may contain amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-directed mutagenesis in vitro or by somatic mutations in vivo), such as in CDRs, particularly in CDR3. However, as used herein, the term "human antibody" is not intended to include antibodies that transplant CDR sequences derived from other mammalian species (e.g., mice) onto human frame sequences. In embodiments, all variable and constant regions may be derived from human immunoglobulin sequences (fully human antibodies).

[0075] The antibodies used in the methods disclosed herein may be recombinant human antibodies. As used herein, the term "recombinant human antibody" includes all human antibodies prepared, expressed, generated, or isolated by recombinant means, such as antibodies expressed using recombinant expression vectors transfected into host cells (described further below), antibodies isolated from a library of recombinant human antibodies (described further below), antibodies isolated from transgenic animals (e.g., mice) containing human immunoglobulin genes (see, for example, Taylor et al., (1992) Nucl. Acids Res. 20:6287-6295), or antibodies prepared, expressed, generated, or isolated by any other means involving splicing human immunoglobulin gene sequences into other DNA sequences. Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. However, in some implementations, such recombinant human antibodies are subjected to in vitro mutagenesis (or, when using transgenic animals targeting human Ig sequences, in vivo somatic cell mutagenesis), so that the amino acid sequences of the VH and VL regions of the recombinant antibody are sequences that, although derived from and associated with human germline VH and VL sequences, may not naturally exist in the human antibody germline library in vivo.

[0076] In some embodiments, the PD-L1 inhibitor is an anti-PD-L1 antibody (e.g., IMC-001) comprising three heavy chain complementarity-determining regions (HCDRs) of a heavy chain variable region (HCVR) containing or consisting of the amino acid sequence of SEQ ID NO:1; and three light chain complementarity-determining regions (LCDRs) of a light chain variable region (LCVR) containing or consisting of the amino acid sequence of SEQ ID NO:2. In some embodiments, the anti-PD-L1 antibody comprises three HCDRs (HCDR1, HCDR2, and HCDR3) and three LCDRs (LCDR1, LCDR2, and LCDR3), wherein HCDR1 comprises or is composed of the amino acid sequence of SEQ ID NO:5; HCDR2 comprises or is composed of the amino acid sequence of SEQ ID NO:6; HCDR3 comprises or is composed of the amino acid sequence of SEQ ID NO:7; LCDR1 comprises or is composed of the amino acid sequence of SEQ ID NO:8; LCDR2 comprises or is composed of the amino acid sequence of SEQ ID NO:9; and LCDR3 comprises or is composed of the amino acid sequence of SEQ ID NO:10. In some embodiments, the anti-PD-L1 antibody comprises HCVR and LCVR, wherein the HCVR comprises or is composed of SEQ ID NO:1, and the LCVR comprises or is composed of SEQ ID NO:2. In some embodiments, the anti-PD-L1 antibody comprises a heavy chain and a light chain, wherein the heavy chain may comprise or consist of the amino acid sequence of SEQ ID NO:3, and the light chain may comprise or consist of the amino acid sequence of SEQ ID NO:4. In this specification, a fully human IgG1 monoclonal antibody comprising a heavy chain having the amino acid sequence of SEQ ID NO:3 and a light chain having the amino acid sequence of SEQ ID NO:4 is referred to as "IMC-001".

[0077] In some embodiments, the anti-PD-L1 antibody may comprise an HCVR having at least 90%, at least 95%, at least 97%, or at least 98% sequence identity with SEQ ID NO:1. In some embodiments, the anti-PD-L1 antibody may comprise an LCVR having at least 90%, at least 95%, at least 97%, or at least 98% sequence identity with SEQ ID NO:2. In some embodiments, the anti-PD-L1 antibody may comprise an HCVR having at least 90%, at least 95%, at least 97%, or at least 98% sequence identity with SEQ ID NO:1 and an LCVR having at least 90%, at least 95%, at least 97%, or at least 98% sequence identity with SEQ ID NO:2. Sequence identity can be measured by methods known in the art (e.g., GAP, BESTFIT, and BLAST).

[0078] In some embodiments, sequence identity can be determined based on regions excluding CDRs (e.g., frame sequences). In some embodiments, the bioequivalence of the IMC-001 antibody is an anti-PD-L1 antibody comprising an HCVR having at least 90%, at least 95%, at least 97%, or at least 98% sequence identity with regions other than HCDR1, HCDR2, and HCDR3 in SEQ ID NO:1. In some embodiments, the bioequivalence of the IMC-001 antibody is an anti-PD-L1 antibody comprising an LCVR having at least 90%, at least 95%, at least 97%, or at least 98% sequence identity with regions other than LCDR1, LCDR2, and LCDR3 in SEQ ID NO:2. In some embodiments, the bioequivalence of IMC-001 is an anti-PD-L1 antibody comprising: an HCVR having at least 90%, at least 95%, at least 97%, or at least 98% sequence identity with the regions of SEQ ID NO:1 excluding HCDR1, HCDR2, and HCDR3; and an LCVR having at least 90%, at least 95%, at least 97%, or at least 98% sequence identity with the regions of SEQ ID NO:2 excluding LCDR1, LCDR2, and LCDR3. In some embodiments, the bioequivalence of the IMC-001 antibody is an anti-PD-L1 antibody comprising an LCVR having at least 90%, at least 95%, at least 97%, or at least 98% sequence identity with the regions of SEQ ID NO:2 excluding LCDR1, LCDR2, and LCDR3, and having amino acids identical to those at positions 3 and 5 of SEQ ID NO:2 from the N-terminus to the C-terminus.

[0079] In some embodiments, the IMC-001 antibody or its bioequivalent can be produced by having the above-described light chain amino acid sequence, particularly in CHO cells where light chain fragmentation is not involved.

[0080] In some embodiments, the bioequivalence of the IMC-001 antibody is an anti-PD-L1 antibody comprising an HCVR containing the amino acid sequence of SEQ ID NO:1 and having no more than 5 amino acid substitutions. In some embodiments, the bioequivalence of IMC-001 is an anti-PD-L1 antibody comprising an LCVR containing the amino acid sequence of SEQ ID NO:2 and having no more than 2 amino acid substitutions. In some embodiments, the bioequivalence of IMC-001 is an anti-PD-L1 antibody comprising: an HCVR containing the amino acid sequence of SEQ ID NO:1 and having no more than 5 amino acid substitutions, and an LCVR containing the amino acid sequence of SEQ ID NO:2 and having no more than 2 amino acid substitutions.

[0081] The antibodies used in the methods disclosed herein may be humanized antibodies. As used herein, the sequence of the term "humanized antibody" differs from that of an antibody derived from a non-human species due to substitutions, deletions, and / or additions of one or more amino acids, making the humanized antibody less likely to induce an immune response and / or induce a lower intensity immune response compared to the non-human species antibody when administered to a human subject. In one embodiment, certain amino acids within the framework and constant domains of the heavy and / or light chains of the non-human species antibody are mutated to produce a humanized antibody. In another embodiment, the constant domain of the human antibody is fused with a variable domain of the non-human species. In yet another embodiment, one or more amino acid residues in one or more CDR sequences of the non-human antibody may be altered to reduce the potential immunogenicity of the non-human antibody when administered to a human subject, wherein the altered amino acid residues are not essential for the immune-specific binding of the antibody to its antigen, or the alteration to the amino acid sequence is a conserved change such that the binding of the humanized antibody to the antigen is not significantly worse than that of the non-human antibody to the antigen. Examples of how to prepare humanized antibodies can be found in U.S. Patent Nos. 6,054,297, 5,886,152 and 5,877,293.

[0082] In this embodiment, the anti-PD-L1 antibody is a fully human IgG antibody that binds to the PD-L1 epitope at a ratio of 10:1. -6 Binding with an affinity of M or lower. In an embodiment, the anti-PD-L1 antibody may be an IgG1 or IgG4 subclass. In an embodiment, a fully human antibody of an IgG class (e.g., IgG1 or IgG4) that binds to the PD-L1 epitope is provided, wherein the antibody comprises a heavy chain variable domain and a light chain variable domain, the heavy chain variable domain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence shown in SEQ ID NO:1, and the light chain variable domain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence shown in SEQ ID NO:2. In one embodiment, a fully human antibody of an IgG class (e.g., IgG1 or IgG4) that binds to a PD-L1 epitope is provided, wherein the antibody comprises a heavy chain variable domain and a light chain variable domain, the heavy chain variable domain comprising CDR1, CDR2, and CDR3 domains having amino acid sequences of SEQ ID NO:5, SEQ ID NO:6, and SEQ ID NO:7, respectively, and the light chain variable domain comprising CDR1, CDR2, and CDR3 domains having amino acid sequences of SEQ ID NO:8, SEQ ID NO:9, and SEQ ID NO:10, respectively.

[0083] Further specific details regarding the anti-PD-L1 antibody disclosed herein can be found in international publication WO 2017 / 132562A1. As described in that document, the IMC-001 antibody comprises the heavy chain variable region sequence of SEQ ID NO:1 and the light chain variable region sequence of SEQ ID NO:2, the entire disclosure of which is incorporated herein by reference.

[0084] Preparation of PD-L1 inhibitors

[0085] Antigen-binding proteins can be prepared using any of a variety of conventional techniques. In embodiments of this disclosure, monoclonal antibodies that bind to PD-L1 are provided. For example, the monoclonal antibody can be purified from cells that naturally express the monoclonal antibody (e.g., from a hybridoma that produces the antibody), or generated in a recombinant expression system using any technique known in the art. See, for example, Monoclonal Antibodies, Hybridomas: A New Dimension in Biological Analyses, Kennet et al. (eds.), Plenum Press, New York (1980); and Antibodies: A Laboratory Manual, Harlow and Land (eds.), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1988).

[0086] Monoclonal antibodies can be produced using any technique known in the art, for example, by immortalizing spleen cells collected from transgenic animals after an immunization procedure has been completed. Spleen cells can be immortalized using any technique known in the art, for example, by fusing them with myeloma cells to create a hybridoma. Myeloma cells used in the hybridoma production fusion procedure preferably do not produce antibodies, have high fusion efficiency, and are enzyme-deficient, preventing them from growing in certain selective media that only support the growth of the desired fusion cells (hybridomas). Examples of cell lines suitable for mouse fusion include Sp-20, P3-X63 / Ag8, P3-X63-Ag8.653, NS1 / 1.Ag 41, Sp210-Ag14, FO, NSO / U, MPC-11, MPC11-X45-GTG1.7, and S194 / 5XX0Bul; examples of cell lines suitable for rat fusion include R210.RCY3, Y3-Ag 1.2.3, IR983F, and 4B210. Other cell lines suitable for cell fusion include U-266, GM1500-GRG2, LICR-LON-HMy2, and UC729-6.

[0087] In this example, the polypeptide is produced by a recombinant DNA method: inserting a nucleic acid sequence (e.g., cDNA) encoding the polypeptide into a recombinant expression vector, and expressing the DNA sequence under conditions that promote expression.

[0088] To recombinantly generate anti-PD-L1 antibodies, the nucleic acid encoding the antibody is isolated and inserted into one or more vectors for further cloning and / or expression in host cells. Such nucleic acids can be readily isolated and sequenced using routine procedures (e.g., using oligonucleotide probes capable of specifically binding to genes encoding the heavy and light chains of the antibody).

[0089] The nucleic acid encoding the anti-PD-L1 antibody (or fragment thereof) disclosed herein can be chemically synthesized. Codon usage can be selected to enhance expression in cells. Such codon usage will depend on the cell type selected. Specific codon usage patterns have been developed for E. coli and other bacteria, as well as mammalian cells, plant cells, yeast cells, and insect cells. See, for example, Mayfield et al., Proc. Natl. Acad. Sci. USA. 2003 100(2):438-42; Sinclair et al., Protein Expr. Purif. 2002(1):96-105; Connell ND. Curr. Opin. Biotechnol. 2001 12(5):446-9; Makrides et al., Microbiol. Rev. 1996 60(3):512-38; and Sharp et al., Yeast. 1991 7(7):657-78.

[0090] General techniques for nucleic acid manipulation have been described, for example, in Sambrook et al., *Molecular Cloning: A Laboratory Manual*, Vols. 1-3, Cold Spring Harbor Laboratory Press, 2nd ed., 1989, or F. Ausubel et al., *Current Protocols in Molecular Biology* (Green Publishing and Wiley-Interscience: New York, 1987), and their periodic updates, which are incorporated herein by reference. DNA encoding polypeptides is operatively linked to suitable transcriptional or translational regulatory elements derived from mammalian, viral, or insect genes. Such regulatory elements include transcription promoters, optional operon sequences controlling transcription, sequences encoding suitable mRNA ribosome binding sites, and sequences controlling the termination of transcription and translation. Furthermore, they include the ability to replicate in the host, typically conferred by the origin of replication, and selection genes that facilitate transformant recognition.

[0091] Recombinant DNA can also contain any type of protein tag sequence that can be used to purify proteins. Examples of protein tags include, but are not limited to, histidine tags, FLAG tags, myc tags, HA tags, or GST tags. Cloning and expression vectors suitable for bacterial, fungal, yeast, and mammalian cell hosts can be found in Cloning Vectors: A Laboratory Manual (Elsevier, NY, 1985).

[0092] The expression construct is introduced into a host cell using a method suitable for the host cell. Various methods for introducing nucleic acids into host cells are known in the art, including but not limited to electroporation; transfection using calcium chloride, rubidium chloride, calcium phosphate, DEAE-glucan, or other substances; microbolite bombardment; liposome transfection; and infection (where the vector is an infectious agent). Suitable host cells include prokaryotes, yeast, mammalian cells, or bacterial cells, as described in more detail below.

[0093] Any expression system known in the art can be used to manufacture the recombinant peptides (e.g., recombinant antibodies) disclosed herein. Typically, host cells are transformed with a recombinant expression vector containing DNA encoding the desired peptide. Suitable host cells include prokaryotes, yeast, or higher eukaryotic cells. Prokaryotes include Gram-negative or Gram-positive organisms such as *Escherichia coli* or bacilli. Higher eukaryotic cells include insect cells and established mammalian-derived cell lines. Host cells suitable for cloning or expressing antibody-encoding vectors include the prokaryotic or eukaryotic cells described herein. For example, antibodies can be produced in bacteria, especially where glycosylation and Fc effector function are not required. For the expression of antibody fragments and peptides in bacteria, see, for example, U.S. Patent Nos. 5,648,237, 5,789,199, and 5,840,523. (See also Charlton, Methods in Molecular Biology, Vol. 248 (BKCLo, ed., Humana Press, Totowa, NJ, (2003), pp. 245-254), which describes the expression of antibody fragments in *E. coli*. After expression, the soluble fraction of the antibody can be isolated from the bacterial cell paste and further purified. Higher eukaryotic cells include insect cells and established mammalian-derived cell lines. Examples of suitable mammalian host cell lines include the COS-7 monkey kidney cell line (ATCC CRL 1651) (Gluzman et al., 1981, Cell 23:175), L cells, 293 cells, C127 cells, 3T3 cells (ATCC CCL 163), Chinese hamster ovary (CHO) cells, HeLa cells, the BHK (ATCC CRL 10) cell line, and the CV1 (ATCC CCL) kidney cell line derived from African green monkeys.) The CV1 / EBNA cell line (70%) is described, as in McMahan et al., 1991, EMBO J.10:2821. Cloning and expression vectors suitable for bacterial, fungal, yeast, and mammalian cell hosts are described by Pouwels et al. (Cloning Vectors: A Laboratory Manual, Elsevier, NY, 1985).

[0094] In some implementations, vertebrate cells can be used as hosts for expressing anti-PD-L1 antibodies or fragments thereof. For example, mammalian cell lines adapted for suspension growth may be useful. Other examples of useful mammalian host cell lines include: SV40-transformed monkey kidney CV1 line (COS-7); human embryonic kidney line (293 or 293 cells, e.g., described in Graham et al., J. GenVirol. 36:59 (1977)); young hamster kidney cells (BHK); mouse Sertoli cells (TM4 cells, e.g., described in Mather, Biol. Reprod. 23:243-251 (1980)); monkey kidney cells (CV1); African green monkey kidney cells (VERO-76); human cervical cancer cells (HELA); canine kidney cells (MDCK); Buffalo rat hepatocytes (BRL 3A); human lung cells (W138); human hepatocytes (HepG2); mouse mammary tumors (MMT 060562); TRI cells, e.g., described in Mather et al., Annals As described in NYAcad.Sci.383:44-68 (1982); MRC 5 cells; and FS4 cells. Other useful mammalian host cell lines include Chinese hamster ovary (CHO) cells, including DHFR-CHO cells (Urlaub et al., Proc. Natl. Acad. Sci. USA 77:4216 (1980)); and myeloma cell lines such as Y0, NSO, and Sp2 / 0. For a review of certain mammalian host cell lines suitable for antibody production, see, for example, Yazaki and Wu, Methods in Molecular Biology, Vol. 248 (BKCLo, ed., Humana Press, Totowa, NJ), pp. 255-268 (2003). Examples of suitable mammalian host cell lines include the COS-7 monkey kidney cell line (ATCC CRL 1651) (Gluzman et al., 1981, Cell 23:175), L cells, 293 cells, C127 cells, 3T3 cells (ATCC CCL 163), Chinese hamster ovary (CHO) cells, HeLa cells, BHK (ATCC CRL 10) cell line, and the CV1 / EBNA cell line derived from the African green monkey kidney cell line CV1 (ATCC CCL 70), as described by McMahan et al., 1991, EMBO J.10:2821.

[0095] Besides prokaryotes, eukaryotic microorganisms (such as filamentous fungi or yeast) are also suitable cloning or expression hosts for antibody-encoding vectors, including fungal and yeast strains whose glycosylation pathways have been humanized, thereby producing antibodies with partially or fully humanized glycosylation patterns. See Gerngross, Nat. Biotech. 22:1409-1414 (2004), and Li et al., 24:210-215 (2006).

[0096] Suitable host cells for expressing glycosylated antibodies also originate from multicellular organisms (invertebrates and vertebrates). Examples of invertebrate cells include plant and insect cells. Several baculovirus strains have been identified that can be used in combination with insect cells, particularly for transfecting fall armyworm (Spodoptera frugiperda) cells.

[0097] Cloning and expression vectors suitable for bacterial, fungal, yeast, and mammalian cell hosts are described by Pouwels et al. (Cloning Vectors: A Laboratory Manual, (Elsevier, NY, 1985)).

[0098] Transformed cells can be cultured under conditions that promote peptide expression, and the peptides can be recovered using standard protein purification procedures. One such purification procedure includes, for example, affinity chromatography on a matrix to which all or part (e.g., the extracellular domain) of PD-L1 is bound. The peptides considered for use herein include substantially homogeneous recombinant mammalian anti-PD-L1 antibody peptides that are substantially free of contaminating endogenous substances.

[0099] Therefore, antibodies can be generated using recombinant methods and compositions, for example, as described in U.S. Patent No. 4,816,567. In one embodiment, an isolated nucleic acid encoding the anti-PD-L1 antibody described herein is provided. Such a nucleic acid may encode an amino acid sequence comprising antibody VL and / or an amino acid sequence comprising antibody VH (e.g., the light chain and / or heavy chain of the antibody). In another embodiment, one or more vectors (e.g., expression vectors) comprising such a nucleic acid are provided. In another embodiment, a host cell comprising such a nucleic acid is provided. In one such embodiment, the host cell comprises (e.g., transformed with): (1) a vector comprising a nucleic acid encoding an amino acid sequence comprising antibody VL and an amino acid sequence comprising antibody VH; or (2) a first vector comprising a nucleic acid encoding an amino acid sequence comprising antibody VL; and a second vector comprising a nucleic acid encoding an amino acid sequence comprising antibody VH. In an embodiment, the host cell is a eukaryotic cell, such as Chinese hamster ovary (CHO) cells or lymphoid cells (e.g., Y0, NSO, Sp20 cells). In one embodiment, a method for generating anti-PD-L1 antibodies is provided, wherein the method includes culturing a host cell containing a nucleic acid encoding an antibody as described above under conditions suitable for antibody expression, and optionally recovering the antibody from the host cell (or host cell culture medium).

[0100] The proteins disclosed herein can also be produced using cellular translation systems. For this purpose, the nucleic acids encoding the polypeptides must be modified to enable in vitro transcription to produce mRNA and to enable cell-free translation of the mRNA in the specific cell-free system used (eukaryotic, such as mammalian or yeast cell-free translation systems, or prokaryotic, such as bacterial cell-free translation systems).

[0101] Furthermore, PD-L1 binding peptides can also be produced through chemical synthesis (e.g., by the method described in Solid Phase Peptide Synthesis, 2nd ed., 1984, The Pierce Chemical Co., Rockford, III). Protein modification can also be achieved through chemical synthesis.

[0102] The polypeptides disclosed herein can be purified using protein isolation / purification methods known in the field of protein chemistry. Non-limiting examples include extraction, recrystallization, salting out (e.g., with ammonium sulfate or sodium sulfate), centrifugation, dialysis, ultrafiltration, adsorption chromatography, ion exchange chromatography, hydrophobic chromatography, normal-phase chromatography, reversed-phase chromatography, gel filtration, gel permeation chromatography, affinity chromatography, electrophoresis, countercurrent partitioning, or any combination thereof. After purification, the polypeptides can be exchanged into different buffers and / or concentrated using any of a variety of methods known in the art, including but not limited to filtration and dialysis.

[0103] The purified polypeptide is preferably at least 85% pure, more preferably at least 95% pure, and most preferably at least 98% pure. Regardless of the specific purity value, the polypeptide is pure enough to be used as a pharmaceutical product. Antigen-binding proteins (e.g., antibodies, antibody fragments, antibody derivatives, antibody mutants, and antibody variants) are polypeptides that bind to PD-L1 (preferably human PD-L1). Antigen-binding proteins include proteins that inhibit the biological activity of PD-L1.

[0104] Antigen-binding proteins can be prepared and screened for desired properties using any of a variety of known techniques. Some techniques involve isolating the nucleic acid of a polypeptide chain (or a portion thereof) encoding the antigen-binding protein of interest (e.g., an anti-PD-L1 antibody) and manipulating that nucleic acid using recombinant DNA techniques. The nucleic acid can be fused with another nucleic acid of interest or altered (e.g., by mutagenesis or other conventional techniques), for example, by adding, deleting, or substituting one or more amino acid residues.

[0105] Single-chain antibodies can be formed by linking heavy and light chain variable domain (Fv region) fragments via amino acid bridges (short peptide linkers) to form a single polypeptide chain. Such single-chain Fvs (scFvs) are prepared by fusing DNA encoding the peptide linker between DNA encoding two variable domain polypeptides (VL and VH). The resulting polypeptides can fold themselves to form antigen-binding monomers, or they can form multimers (e.g., dimers, trimers, or tetramers), depending on the length of the flexible linker between the two variable domains (Kortt et al., 1997, Prot. Eng. 10:423; Kortt et al., 2001, Biomol. Eng. 18:95-108). By combining different VL and VH-containing polypeptides, multimer scFvs binding to different epitopes can be formed (Kriangkum et al., 2001, Biomol. Eng. 18:31-40). Techniques developed for generating single-chain antibodies include those described in the following: U.S. Patent No. 4,946,778; Bird, 1988, Science 242:423; Huston et al., 1988, Proc. Natl. Acad. Sci. USA 85:5879; Ward et al., 1989, Nature 334:544; de Graaf et al., 2002, Methods Mol. Biol. 178:379-87.

[0106] Techniques for deriving antibodies from different subclasses or isotypes from an antibody of interest are known, known as subclass conversion. Thus, for example, IgG antibodies can be derived from IgM antibodies, and vice versa. Such techniques can prepare new antibodies that possess the antigen-binding properties of a given antibody (parental antibody) but exhibit biological characteristics associated with a different antibody isotype or subclass than the parent antibody. Recombinant DNA technology can be employed. In such procedures, clonal DNA encoding a specific antibody polypeptide can be used, for example, DNA encoding a constant domain of the desired isotype antibody (Lantto et al., 2002, Methods Mol. Biol. 178:303-16). Furthermore, if IgG4 is desired, it is also desirable to introduce point mutations (CPSCP→CPPCP) into the hinge region (Bloom et al., 1997, Protein Science 6:407) to mitigate the tendency to form inter-chain disulfide bonds, which may lead to heterogeneity in IgG4 antibodies.

[0107] Pharmaceutical Compositions and Administration

[0108] According to another aspect of this disclosure, pharmaceutical compositions comprising a PD-L1 inhibitor as neoadjuvant therapy are provided, as described herein. Such pharmaceutical compositions can be formulated with suitable pharmaceutically acceptable carriers, excipients, buffers, and other agents that provide suitable transport, delivery, tolerability, etc. A variety of suitable formulations can be found in all formularies known to medicinal chemists: Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA. These formulations include, for example, acids, pastes, ointments, gels, waxes, oils, lipids, and lipid-containing (cationic or anionic) vesicles (e.g., LIPOFECTIN). TM DNA conjugates, anhydrous absorbent pastes, oil-in-water and water-in-oil emulsions, polyethylene glycol emulsions (polyethylene glycol of various molecular weights), semi-solid gels, and semi-solid mixtures containing polyethylene glycol. See also Powell et al., “Compendium of excipients for parenteral formulations” PDA, J Pharm Sci Technol 52:238-311 (1998).

[0109] The dosage of a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody) may vary depending on the age and size of the subject to be administered, the target disease, the condition, the route of administration, etc. When the PD-L1 inhibitor of this disclosure is used to treat or inhibit the growth of gastric cancer, esophageal cancer, or liver cancer, the PD-L1 inhibitor may be administered in single or multiple doses at a dose of about 0.1 to about 100 mg / kg body weight. The frequency and duration of treatment may be adjusted according to the severity of the condition. In some embodiments, the PD-L1 inhibitor of this disclosure may be administered at an initial dose of at least about 0.1 mg to about 800 mg, about 1 to about 1000 mg, about 1 to about 800 mg, about 5 to about 500 mg, or about 10 to about 400 mg. In some embodiments, a second or more subsequent doses of the PD-L1 inhibitor may be administered after the initial dose, in an amount that may be substantially the same as or less than the initial dose, wherein the subsequent doses are given at the following intervals: at least 1 to 3 days; at least 1 week; at least 2 weeks; at least 3 weeks; at least 4 weeks; at least 5 weeks; at least 6 weeks; at least 7 weeks; at least 8 weeks; at least 9 weeks; at least 10 weeks; at least 12 weeks; or at least 14 weeks. In some embodiments, subsequent doses of the PD-L1 inhibitor after the initial dose are preferably given at intervals of at least 1 week, at least 2 weeks, at least 3 weeks, or at least 4 weeks.

[0110] Various delivery systems are known and can be used to administer the pharmaceutical compositions disclosed herein, such as encapsulation in liposomes, microparticles, microcapsules, recombinant cells capable of expressing mutant viruses, and receptor-mediated endocytosis (see, for example, Wu et al., (1987) J. Biol. Chem. 262:4429-4432). Methods of administration include, but are not limited to, intradermal, transdermal, intramuscular, intravenous, subcutaneous, intranasal, epidural, and oral routes. The compositions can be administered via any convenient route, such as by infusion or bolus injection, absorption through epithelial or mucocutaneous linings (e.g., oral mucosa, rectal and intestinal mucosa), and can be administered with other bioactive agents. The pharmaceutical compositions can also be delivered in vesicles, particularly liposomes (see, for example, Langer (1990) Science 249:1527-1533).

[0111] This document also envisions the use of nanoparticles to deliver the PD-L1 inhibitor disclosed herein. Antibody-conjugated nanoparticles can be used for therapeutic and diagnostic applications. Antibody-conjugated nanoparticles and methods of their preparation and use are described in detail in Arruebo, M. et al., 2009, “Antibody-conjugated nanoparticles for biomedical applications,” J. Nanomat., Vol. 2009, Article ID 439389, page 24. Nanoparticles can be developed and conjugated with antibodies contained in pharmaceutical compositions to target cells. Nanoparticles for drug delivery have also been described, for example, in US 8257740 or US 8246995.

[0112] In some cases, the drug composition can be delivered using a controlled-release system. In one embodiment, a pump can be used. In another embodiment, a polymer material can be used. In yet another embodiment, the controlled-release system can be placed near the target of the composition, thus requiring only a portion of the systemic dose.

[0113] Injectable formulations may include dosage forms for intravenous, subcutaneous, intracranial, and intramuscular injection, infusion, etc. These injectable formulations can be prepared using known methods. In one embodiment, the injectable formulation may be a histidine-based solution.

[0114] The pharmaceutical compositions disclosed herein can be delivered subcutaneously or intravenously using standard needles and syringes. Furthermore, for subcutaneous delivery, pen-type delivery devices are readily applicable in delivering the pharmaceutical compositions of this disclosure. Such pen-type delivery devices can be reusable or disposable. Reusable pen-type delivery devices typically use replaceable cartridges containing the pharmaceutical composition. Once all the pharmaceutical composition in the cartridge has been administered and the cartridge is empty, the empty cartridge can be easily discarded and replaced with a new cartridge containing the pharmaceutical composition. The pen-type delivery device can then be reused. In disposable pen-type delivery devices, there are no replaceable cartridges. Instead, the disposable pen-type delivery device is pre-loaded with the pharmaceutical composition, which is contained in a reservoir within the device. Once the pharmaceutical composition in the reservoir is used up, the entire device can be discarded.

[0115] Advantageously, the above-described oral or parenteral pharmaceutical compositions are prepared in unit-dose dosage forms suitable for the dosage of the active ingredient. Such unit-dose dosage forms include, for example, tablets, pills, capsules, injections (ampoules), suppositories, etc. The amount of antibody contained in each unit-dose dosage form is typically from about 5 to about 1000 mg, for example, from about 5 to about 600 mg, from about 5 to about 350 mg, or from about 10 to about 300 mg.

[0116] In some embodiments, this disclosure provides pharmaceutical compositions or formulations comprising a therapeutically effective amount of a PD-L1 inhibitor (e.g., IMC-001 or its bioequivalence) and a pharmaceutically acceptable delivery vehicle. Non-limiting examples of pharmaceutical compositions comprising the anti-PD-L1 antibody provided herein that can be used in the context of this disclosure are disclosed in US2019 / 0040137.

[0117] Furthermore, according to another aspect of this disclosure, a kit is provided that contains a PD-L1 inhibitor (e.g., IMC-001 or its bioequivalent) for the therapeutic use described herein. Kits typically include a label indicating the intended use of the kit contents and a user manual. As used herein, the term "label" includes any written or recorded material provided on, inside, or accompanying the kit, or otherwise provided with the kit. In embodiments, a kit is provided for treating patients with upper gastrointestinal cancers (e.g., gastric, esophageal, or liver cancer) comprising: (a) a therapeutically effective amount of a PD-L1 inhibitor (e.g., IMC-001 or its bioequivalent); and (b) a user manual for using the PD-L1 inhibitor in any of the methods described herein.

[0118] Dosage regimen and dosage

[0119] In some embodiments, the methods of this disclosure may include administering a therapeutically effective amount of a PD-L1 inhibitor (e.g., an IMC-001 antibody or its bioequivalence) to a subject's tumor at least once, such as once, twice, three times, four times, five times, six times, seven times, eight times, nine times, or ten times. For example, the treatment dosing regimen may include administering one or more doses of the PD-L1 inhibitor to the subject at intervals of approximately 1 day, approximately 2 days, approximately 3 days, approximately 4 days, approximately 5 days, approximately 6 days, approximately 1 week, approximately 2 weeks, approximately 3 weeks, approximately 4 weeks, approximately 5 weeks, approximately 6 weeks, approximately 7 weeks, approximately 8 weeks, approximately 12 weeks, approximately 1 month, approximately 2 months, approximately 3 months, or approximately 4 months or longer, or as needed, as long as a therapeutic response is achieved.

[0120] In some embodiments, the PD-L1 inhibitor is administered for at least one treatment cycle, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 treatment cycles. Methods according to this aspect include administering one or more cycles of neoadjuvant therapy to a subject in need, optionally followed by one or more cycles of adjuvant therapy. Each treatment cycle includes administering a PD-L1 inhibitor (e.g., IMC-001 or its bioequivalent) once, twice, three times, four times, five times, six times, seven times, eight times, nine times, ten times, or more.

[0121] In one embodiment, the neoadjuvant therapy involves administering a PD-L1 inhibitor once, twice, three times, four times, five times, or six times prior to surgical resection of the tumor. In another embodiment, the neoadjuvant therapy involves administering a PD-L1 inhibitor two to six times prior to surgical resection of the tumor. In yet another embodiment, the neoadjuvant therapy involves administering a total of two doses (two treatment cycles) of a PD-L1 inhibitor prior to surgical resection of the tumor. In yet another embodiment, the neoadjuvant therapy involves administering a PD-L1 inhibitor at least twice (two treatment cycles) and at most six times (six treatment cycles) prior to surgical resection of the tumor. In yet another embodiment, the neoadjuvant therapy involves administering a PD-L1 inhibitor once every approximately one week, approximately two weeks, or approximately three weeks. In yet another embodiment, the neoadjuvant therapy involves administering a PD-L1 inhibitor at intervals of 10 to 18 days, preferably 14 days.

[0122] In some embodiments, the PD-L1 inhibitor is administered at a dose of 0.1, 1, 0.3, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 mg / kg of patient body weight. In some embodiments, the PD-L1 inhibitor is administered at a dose of 20 mg / kg at 10-18 day intervals (two administration cycles) prior to surgical resection of the tumor.

[0123] The amount of a PD-L1 inhibitor (e.g., IMC-001 or its bioequivalent) administered to a subject according to the methods disclosed herein is generally a therapeutically effective amount. As used herein, the term "therapeuticly effective amount" refers to the amount of a PD-L1 inhibitor administered as neoadjuvant therapy prior to planned surgery for the treatment of gastric, esophageal, or hepatic cancer that results in at least one of the following outcomes: (a) inhibition of tumor proliferation or increased tumor necrosis, tumor shrinkage, and / or tumor disappearance; (b) reduction of the severity or duration of symptoms or indications of cancer (e.g., tumor lesions); (c) delay of tumor proliferation and development; (d) inhibition of tumor metastasis; (e) prevention of tumor proliferation and recurrence; and / or (f) increased survival in a cancer subject, each of which is compared to a subject treated with surgical resection alone or a untreated subject.

[0124] In some embodiments, the therapeutically effective amount of a PD-L1 inhibitor (e.g., IMC-001 or its bioequivalent) may be an antibody of about 0.05 mg to about 1000 mg, about 1 mg to about 800 mg, about 5 mg to about 600 mg, about 10 mg to about 550 mg, about 50 mg to about 400 mg, about 75 mg to about 350 mg, or about 100 mg to about 300 mg. For example, in several embodiments, the amount of the PD-L1 inhibitor is about 0.05 mg, about 0.1 mg, about 1.0 mg, about 1.5 mg, about 2.0 mg, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 19 mg, etc. 0mg, approximately 200mg, approximately 210mg, approximately 220mg, approximately 230mg, approximately 240mg, approximately 250mg, approximately 260mg, approximately 270mg, approximately 280mg, approximately 290mg, approximately 300mg, approximately 310mg, approximately 320mg, approximately 330mg, approximately 340mg, approximately 350mg, approximately 360mg, approximately 370mg, approximately 380mg, approximately 390mg, approximately 400mg, approximately 410mg, approximately 420mg, approximately 430mg, approximately 440mg, approximately 450mg, approximately 460mg g, approximately 470mg, approximately 480mg, approximately 490mg, approximately 500mg, approximately 510mg, approximately 520mg, approximately 530mg, approximately 540mg, approximately 550mg, approximately 560mg, approximately 570mg, approximately 580mg, approximately 590mg, approximately 600mg, approximately 610mg, approximately 620mg, approximately 630mg, approximately 640mg, approximately 650mg, approximately 660mg, approximately 670mg, approximately 680mg, approximately 690mg, approximately 700mg, approximately 710mg, approximately 720mg, approximately 730mg, Approximately 740mg, approximately 750mg, approximately 760mg, approximately 770mg, approximately 780mg, approximately 790mg, approximately 800mg, approximately 810mg, approximately 820mg, approximately 830mg, approximately 840mg, approximately 850mg, approximately 860mg, approximately 870mg, approximately 880mg, approximately 890mg, approximately 900mg, approximately 910mg, approximately 920mg, approximately 930mg, approximately 940mg, approximately 950mg, approximately 960mg, approximately 970mg, approximately 980mg, approximately 990mg, or approximately 1000mg.

[0125] The amount of PD-L1 inhibitor (e.g., IMC-001 or its bioequivalent) contained in a single dose can be expressed in milligrams of antibody per kilogram of subject body weight (i.e., mg / kg). In some embodiments, the PD-L1 inhibitor used in the methods disclosed herein can be administered to the subject at a dose of about 0.0001 to about 100 mg / kg of subject body weight. In some embodiments, the anti-PD-L1 antibody can be administered to the subject at a dose of about 0.1 mg / kg to about 30 mg / kg of subject body weight. In some embodiments, the methods of this disclosure include administering a PD-L1 inhibitor (e.g., anti-PD-L1 antibody IMC-001 or its bioequivalent) at doses of about 1 mg / kg to 30 mg / kg, 5 mg / kg to 30 mg / kg, 10 mg / kg to 30 mg / kg, 15 mg / kg, or 20 mg / kg of subject body weight.

[0126] In some embodiments, the individual dose of a PD-L1 inhibitor (e.g., IMC-001 or its bioequivalent) administered to a patient may be lower than the therapeutically effective dose, i.e., a subtherapeutic dose. For example, if the therapeutically effective dose of a PD-L1 inhibitor includes 20 mg / kg, then a subtherapeutic dose includes amounts below 20 mg / kg, such as 18 mg / kg, 16 mg / kg, 14 mg / kg, 12 mg / kg, or 10 mg / kg. As defined herein, a “subtherapeutic dose” refers to an amount of PD-L1 inhibitor that does not produce a therapeutic effect on its own. However, in some embodiments, multiple subtherapeutic doses of the PD-L1 inhibitor are administered to achieve a combined therapeutic effect in the subject.

[0127] In some embodiments, each dose contains 0.1 to 30 mg / kg (e.g., 5 mg / kg, 10 mg / kg, 15 mg / kg, 20 mg / kg, 20 mg / kg, or 25 mg / kg) of a PD-L1 inhibitor (e.g., IMC-001 or its bioequivalent), based on the patient's weight. In some other embodiments, each dose contains 5 to 600 mg of a PD-L1 inhibitor, such as 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 40 mg, 45 mg, 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 400 mg, 500 mg, or 600 mg of a PD-L1 inhibitor.

[0128] In one embodiment, a therapeutically effective dose of a PD-L1 inhibitor (e.g., IMC-001 or its bioequivalent) is 20 mg / kg, administered intravenously as neoadjuvant therapy prior to planned surgery for gastric, esophageal, or hepatic cancer. In some embodiments, another therapeutically effective dose of a PD-L1 inhibitor (e.g., IMC-001 or its bioequivalent) is 20 mg / kg, administered intravenously postoperatively as adjuvant therapy.

[0129] In some embodiments, surgical resection of the tumor is performed 11 days or 84 days after the last administration of the PD-L1 inhibitor. In some embodiments, surgical resection of the tumor is performed between 11 and 84 days after the last administration of the PD-L1 inhibitor. In some embodiments, surgical resection of the tumor is performed between 11 and 42 days after the last administration of the PD-L1 inhibitor. In some embodiments, surgical resection of the tumor is performed between 18 and 35 days or between 21 and 35 days after the last administration of the PD-L1 inhibitor.

[0130] In some implementations, the patient is given an anti-PD-L1 antibody at a dose of 20 mg / kg and at intervals of 10 to 18 days or every 2 weeks prior to surgical resection of the tumor, and surgical resection of the tumor is performed between 11 and 42 days after the last administration of the anti-PD-L1 antibody.

[0131] All patents and references cited in this article are incorporated herein by reference in their entirety.

[0132] The present disclosure will be described in more detail below through the following embodiments. However, the following embodiments are for illustrative purposes only, and the scope of the present disclosure is not limited thereto.

[0133] Example

[0134] Example 1. Clinical trial of IMC-001 as a neoadjuvant therapy for resectable gastric, esophageal, and liver cancer.

[0135] This study is a phase 2 multi-cohort trial of the immune checkpoint inhibitor IMC-001 as neoadjuvant therapy for resectable localized gastric adenocarcinoma (GC), esophageal squamous cell carcinoma (ESCC), or hepatocellular carcinoma (HCC). IMC-001 is a fully human anti-PD-L1 recombinant monoclonal antibody of the IgG1 type, comprising a heavy chain and a light chain. The heavy chain contains the amino acid sequence shown in SEQ ID NO:3, and the light chain contains the amino acid sequence shown in SEQ ID NO:4. The amino acid sequence information of IMC-001 is shown in Table 1 below.

[0136] [Table 1]

[0138] IMC-001 is H6B1L-EM, a variant of the wild-type parental antibody H6B1L, produced in CHO cells. For more details, see international publication number WO 2017 / 132562A1.

[0139] The study included the following cohorts: a cohort of patients with resectable gastric cancer; a cohort of patients with resectable esophageal cancer; and a cohort of patients with resectable hepatocellular carcinoma.

[0140] Research Objective

[0141] The aim of this study was to evaluate the clinical activity of IMC-001 as a neoadjuvant therapy in patients with resectable lesions of gastric, esophageal, and hepatocellular carcinoma.

[0142] Basic principles

[0143] Neoadjuvant immunotherapy offers potential biological benefits, including less immunosuppressive tumor microenvironment (TME), lower disease burden, and antitumor immune initiation in the presence of a primary tumor. Digestive system cancers (including gastroesophageal and hepatocellular carcinoma) are a class of cancers for which immune checkpoint inhibitors (ICIs) have shown survival benefits in palliative care. IMC-001, a fully human anti-PD-L1 recombinant monoclonal antibody that retains the Fc effector to stimulate ADCC, demonstrated good safety in a phase 1 study (n=15) when administered intravenously at doses of up to 20 mg / kg every 2 weeks, and showed promising initial efficacy in patients with previously treated solid tumors (1 partial response, 4 stable disease) (Keam B et al., Invest New Drugs 2021; 39:1624-32).

[0144] Study endpoints

[0145] The primary endpoint of this study was to evaluate the major pathological response rate in resected tissue following preoperative administration of the immune checkpoint inhibitor IMC-001. Secondary endpoints included assessing the feasibility and safety of postponing planned surgery when IMC-001 was used as preoperative therapy; assessing the R0 resection rate; assessing clinical tumor response rate according to RECIST v1.1; assessing clinical disease control rate according to RECIST v1.1; assessing progression-free survival; assessing recurrence-free survival; assessing overall survival; and assessing cancer progression / recurrence rate and cancer progression / recurrence patterns. Additionally, an exploratory objective of this study was to obtain samples (tissue, blood, and stool) from participants before and after administration of the immune checkpoint inhibitor IMC-001 to establish a biomarker cohort, and to perform immunoprofiling and genomic analyses to identify biomarkers that could predict treatment efficacy.

[0146] Research Design

[0147] Subjects meeting the inclusion / exclusion criteria were assigned to the corresponding cohorts for gastric cancer, esophageal cancer, and hepatocellular carcinoma. Within each cancer cohort, subjects received two cycles of IMC-001 (1 cycle = 2 weeks) as pre-treatment, followed by radical surgery and subsequent follow-up. After radical surgery, adjuvant therapy was administered as appropriate based on disease stage and according to standard practice guidelines.

[0148] The study targeted patients who had not yet received treatment for resectable localized gastric cancer, esophageal cancer, or hepatocellular carcinoma. A total of 48 patients were included in the study (16 patients in each cancer cohort).

[0149] Neoadjuvant therapy

[0150] In the absence of unacceptable toxicity, clinical cancer progression, or withdrawal of consent, IMC-001 may be administered twice every two weeks at 20 mg / kg as a pre-treatment prior to surgical resection.

[0151] Biomarker sample collection

[0152] Tumor tissue, blood, and stool were collected before IMC-001 was administered. Blood and stool were collected after the first and second cycles of administration. Excised tumor tissue, blood, and stool were collected after surgery.

[0153] Follow-up

[0154] From the date of surgery, every three months for the first two years, every six months for the following three years, and in the event of cancer recurrence, blood, stool, and tumor tissue were collected and observed (only in the case of recurrence).

[0155] Subjects

[0156] This study included 48 patients with gastric adenocarcinoma, esophageal squamous cell carcinoma, or hepatocellular carcinoma (16 patients in each cancer cohort), all of whom were capable of radical surgical resection.

[0157] Inclusion criteria

[0158] Disease inclusion criteria

[0159] 1) Histologically confirmed localized gastric adenocarcinoma, esophageal squamous cell carcinoma, hepatocellular carcinoma, or hepatocellular carcinoma clinically diagnosed according to the American Association for the Study of Liver Diseases (AASLD) guidelines. However, biopsy is not performed for hepatocellular carcinoma that can be clinically diagnosed according to the AASLD guidelines.

[0160] 2) Gastric adenocarcinoma, esophageal squamous cell carcinoma, or hepatocellular carcinoma, where radical resection is expected.

[0161] - Gastric adenocarcinoma: Clinical stage ≥T2 or positive lymph node metastasis (AJCC 8th edition)

[0162] - Esophageal squamous cell carcinoma: Clinical stage ≥T1b or positive lymph node metastasis (AJCC 8th edition)

[0163] - Hepatocellular carcinoma: A single hepatocellular carcinoma confined to the liver, or three or fewer hepatocellular carcinomas confined to the liver. Even if there is portal vein, hepatic vein, or bile duct invasion, there must be no invasion of the main portal vein.

[0164] 3) Hematological, blood chemistry, and major organ function requirements must meet the following criteria (confirmed within 7 days prior to the first dose of the study drug).

[0165] - Absolute neutrophil count ≥1000 / μL

[0166] - Platelet count ≥75000 / μL

[0167] - Serum total bilirubin ≤1.5x Upper Limit of Normal (ULN) (For subjects with Gilbert's syndrome, total bilirubin ≤3.0x ULN)

[0168] - Aspartate aminotransferase (AST) or alanine aminotransferase (ALT) ≤ 2.5x ULN; alkaline phosphatase ≤ 2.5x ULN

[0169] - Serum creatinine ≤1.5x upper limit of normal, or creatinine clearance ≥50 mL / min (creatinine clearance is first calculated according to sex using the following Cockcroft-Gault formula. If the value is less than 50 mL / min, a 24-hour urine collection test is performed. Here, subjects with a creatinine clearance of 50 mL / min or higher are eligible for enrollment).

[0170] [Male Creatinine Clearance Rate]

[0171]

[0172] [Female creatinine clearance rate]

[0173]

[0174] - Urine protein-creatinine ratio (UPC) ≤ 1 (However, subjects with UPC > 1 may be admitted if the 24-hour urine protein test shows < 2g).

[0175] - In addition, for hepatocellular carcinoma, subjects must have Child-Pugh A (see Table 4) and grade 0 encephalopathy (see Table 3), and have confirmed adequate liver function (see Table 2).

[0176] [Table 2] score a. Select the higher score: the extension time (seconds) or INR result relative to the control.

[0177] [Table 3] Brain disease classification

[0178] [Table 4] Classification Fraction Child-Pugh classification 5-6 A 7-9 B 10-15 C

[0179] 4) Measurable or evaluable lesions according to the RECIST version 1.1 criteria for evaluating the efficacy of treatment in solid tumors (see Eisenhauer EA, Therasse P, Bogaerts J, Schwartz LH, Sargent D, Ford R et al., New response evaluation criteria in solid tumors: revised RECIST guideline (Version 1.1). Eur J Cancer. 2009; 45:228 247).

[0180] 5) The study must be able to provide tumor tissue samples suitable for biomarker analysis (for hepatocellular carcinoma, enrollment is possible even if no tumor tissue sample is available before administration of the study drug).

[0181] General inclusion criteria

[0182] 6) Age 19 or above

[0183] 7) Eastern Cooperative Oncology Group (ECOG) performance status 0 to 1 (see Table 5)

[0184] [Table 5]

[0185] 8) Informed consent form must be signed.

[0186] 9) For fertile male and female subjects, they must be able to adhere to appropriate contraception during treatment and for up to 3 months after the last dose of study drug.

[0187] Exclusion criteria

[0188] Subjects who meet any of the following criteria will be excluded from clinical trials.

[0189] Tumor-related exclusion criteria

[0190] 1) Unresectable or metastatic disease

[0191] 2) Prior treatment for gastric adenocarcinoma, esophageal squamous cell carcinoma, or hepatocellular carcinoma. However, for hepatocellular carcinoma, even if the subject has received prior treatment for the local lesion, if more than 6 months have passed since treatment, the treated area has progressed, or a new lesion has appeared outside the previously treated area, and radical resection is possible, and the subject meets other inclusion / exclusion criteria, the subject will still be eligible for clinical trial enrollment.

[0192] 3) Patients with a history of other cancers within 3 years prior to starting the trial treatment. However, enrollment of patients with cancers unlikely to affect prognosis (such as carcinoma in situ or papillary thyroid carcinoma) may be decided by the investigator at their discretion.

[0193] 4) History of hepatic encephalopathy

[0194] 5) Clinically significant ascites, defined as follows:

[0195] - Ascites was found during physical examination during screening, or

[0196] -Previous history of ascites requiring treatment and ongoing prevention, or current ascites requiring treatment.

[0197] Exclusion criteria for investigational drugs

[0198] 6) A history of active autoimmune disease requiring systemic treatment (i.e., disease modifiers, corticosteroids, or immunosuppressants) within the past two years. Replacement therapies (e.g., physiological corticosteroid replacement therapy due to dysfunction of thyroid hormones or insulin, adrenal glands, or pituitary gland) are not considered a form of systemic treatment and are permitted.

[0199] 7) Subjects diagnosed with immunodeficiency, or who received chronic systemic steroid therapy (prednisone or equivalent doses exceeding 10 mg / day) or any other form of immunosuppressive therapy within 7 days prior to the first dose of the study drug.

[0200] 8) Subjects with a history of (non-infectious) interstitial pneumonia requiring steroid treatment or currently suffering from interstitial pneumonia.

[0201] 9) Prior treatment with anti-PD-1, anti-PD-L1, anti-PD-L2, anti-CTLA-4 antibodies or other antibodies or drugs that specifically target T cell co-stimulation or checkpoint pathways.

[0202] 10) Known to have severe hypersensitivity or allergic reactions to recombinant proteins (including monoclonal antibodies).

[0203] General exclusion criteria

[0204] 11) Active infections requiring systemic treatment

[0205] 12) There is a history or current evidence of any medical condition, treatment, or laboratory abnormality that the investigator believes would prevent a participant from participating throughout the duration of the clinical trial or would not be in the participant's best interest in participation, or that may affect the outcome of the clinical trial.

[0206] 13) Fertility-appropriate female subjects who had a positive urine or blood pregnancy test within 7 days prior to the first dose of the study drug.

[0207] 14) Female subjects who are pregnant or breastfeeding, or who plan to become pregnant or give birth within up to 90 days during the clinical trial and after the last dose of the investigational drug.

[0208] 15) A history or current evidence of symptomatic congestive heart failure (i.e., New York Heart Association Class II) or clinically significant arrhythmias (requiring treatment with antiarrhythmic drugs other than beta-blockers or digoxin), and / or conduction abnormalities (excluding atrial fibrillation and paroxysmal ventricular tachycardia), active coronary artery disease, unstable angina within 6 months prior to the start of the clinical trial, new-onset angina within 3 months prior to enrollment in the clinical trial, or myocardial infarction within 6 months prior to enrollment in the clinical trial.

[0209] 16) Known history of human immunodeficiency virus (HIV) infection (HIV1 / 2 antibody)

[0210] 17) Subjects with known active hepatitis B (detectable hepatitis B surface antigen HBsAg or HBV DNA) or hepatitis C (detectable HCV RNA). However, subjects with hepatitis B may be admitted if HBV DNA is <500 IU / mL (or 2500 copies / mL) at screening. Subjects with detectable HBsAg or detectable HBV DNA should be managed according to treatment guidelines. Subjects receiving antiviral drugs at screening must receive at least 2 weeks of treatment prior to enrollment and must continue treatment for 6 months after treatment with study drugs. Patients who are HCV antibody positive will only be admitted if HCV RNA is negative in PCR testing (however, subjects with hepatocellular carcinoma will be admitted regardless of whether their HCV RNA is positive).

[0211] 18) History of allogeneic tissue / solid organ transplantation

[0212] 19) Received a live vaccine within 28 days prior to the first dose of the investigational drug.

[0213] Experimental treatment (preoperative therapy using IMC-001)

[0214] Methods for preparing and administering investigational drugs

[0215] IMC-001 is provided in injectable form in 10 mL sterile glass vials at concentrations of 10 mg / mL and 60 mg / mL in histidine solution. IMC-001 is diluted in 250 mL of 0.9% sodium chloride and administered via intravenous infusion over 60 minutes (±10 minutes). During IMC-001 administration, subjects should be closely monitored for any infusion-related reactions. Preoperative administration is permitted, and antihistamines, antipyretics, and / or analgesics (e.g., acetaminophen) may be administered preoperatively at the investigator's discretion. If infusion-related reactions occur, subjects should be managed according to treatment guidelines. Vital signs (blood pressure, pulse rate, respiratory rate, and temperature) should be measured within 60 minutes prior to infusion. If clinically indicated, vital signs should be measured every 15 minutes (±10 minutes) during infusion or 30 minutes (±10 minutes) after infusion. For subjects who experienced infusion-related reactions during the first infusion, vital signs were measured during the infusion and 30 minutes (±10 minutes) after the infusion.

[0216] Initial dose and dosing regimen

[0217] In this clinical trial, the dose of IMC-001 was 20 mg / kg, which was determined in the ongoing Phase 1 clinical trial and is the recommended dose (RP2D) for the Phase 2 clinical trial. It was administered intravenously on day 1 (± day 4) of each 14-day cycle. Dosage calculations were based on weight assessed at baseline. If a subject's weight differed from the weight used in the previous dose calculation by less than 10%, the dose did not need to be recalculated. All doses were rounded to the nearest milligram (mg) according to the standards of the clinical trial site.

[0218] Total application duration

[0219] Two cycles of IMC-001 were administered prior to surgery unless the subject discontinued treatment prematurely due to unacceptable toxicity or cancer progression, or it was determined that continued treatment would be harmful, or unless informed consent was withdrawn.

[0220] Operation:

[0221] Surgery was performed within four weeks of completion of the second cycle of IMC-001 preoperative therapy (between days 11 and 42 from the date of IMC-001 administration [day 1]). However, the end of the prior therapy visit was earlier than the surgery date. At this time, the subject had a white blood cell count greater than 3,000 / μL, a platelet count greater than 75,000 / μL, no grade 2 or higher toxicity due to study drug treatment, and a clinically suitable condition for major surgery. Surgical procedures were performed according to institutional treatment guidelines for gastric, esophageal, and hepatocellular carcinoma. Open surgery was considered the standard approach. Laparoscopic and robot-assisted surgeries were performed only in consultation with the principal investigator and performed by a surgical team with extensive expertise, where laparoscopic and robot-assisted surgeries were considered comparable to open surgery in terms of morbidity, mortality, and oncological outcomes. During the initial evaluation phase, the surgical team may consider laparoscopic surgery for clinical trials after reviewing evidence of tumor extent, resectability, and distant metastasis.

[0222] Excision surgery

[0223] The extent of surgical resection is determined based on the tumor's location, size, and progression. The resectability of cancer is determined by the attending surgeon during the procedure. The following situations may be considered reasons for inoperability:

[0224] -Remote transfer

[0225] - Tumor invasion of major vascular structures and / or other organs

[0226] - Other situations where radical resection is deemed impossible

[0227] Generally, if complete resection (R0) is not feasible, the surgical team decides to perform a non-radical surgery.

[0228] Lymph node dissection

[0229] The extent of lymph node dissection is determined according to institutional treatment guidelines for gastric cancer, esophageal cancer, and hepatocellular carcinoma.

[0230] Postoperative treatment

[0231] Adjuvant chemotherapy following radical resection and treatment of patients who have undergone non-radical resection or are not candidates for resection are administered in accordance with institutional guidelines.

[0232] Contraindicated concomitant medications and treatments

[0233] The following drugs and treatments are prohibited during clinical trials (except for those used to treat adverse events related to the investigational drug).

[0234] - During this clinical trial, any anticancer therapy other than the investigational drug used to treat cancer (including chemotherapy, hormone therapy, immunotherapy, radiotherapy, biological anticancer therapy, or traditional herbal remedies) during the period of prior anticancer therapy administration.

[0235] - Any other investigational treatments used in this clinical trial within 28 days prior to the start of administration of the investigational drug and during administration of the investigational drug.

[0236] - Live attenuated vaccines (e.g., within 28 days before the start of administration of the study drug, during administration of the study drug, and 5 months after the last dose of the study drug) may be administered intravenously (e.g., within 28 days before the start of administration of the study drug, during administration of the study drug, and 5 months after the last dose of the study drug). ).

[0237] - Systemic immunosuppressants (cyclophosphamide, azathioprine, methotrexate, thalidomide, etc.).

[0238] - Systemic corticosteroids: Generally, corticosteroids at doses greater than or equal to 10 mg / day of prednisone are not permitted. Temporary use of corticosteroids exceeding 10 mg / day is permissible if medically necessary, such as for managing symptoms of infusion reactions or adverse events like intravenous inflammatory response (irAE), or before computed tomography (CT) imaging in subjects with a history of allergy to intravenous contrast agents. If the corticosteroid dose cannot be reduced to 10 mg / day or a lower prednisone dose within 12 weeks of starting steroids, the investigator should immediately initiate a discussion regarding whether to continue treatment with the investigational drug in the subject.

[0239] Permitted concomitant medications and treatments

[0240] The following medications and treatments are permitted.

[0241] - Oral contraceptives

[0242] Hormone replacement therapy

[0243] - Inactivated influenza vaccine

[0244] Medroxyprogesterone acetate as an appetite stimulant

[0245] - Topical, ophthalmic, intra-articular, intranasal, or inhaled corticosteroids

[0246] - Adrenal replacement steroids, prednisone <10mg / day

[0247] - Corticosteroids used temporarily for prevention (e.g., contrast agent allergy).

[0248] The use of other medications required for subject management is at the investigator's discretion. If nausea, vomiting, or diarrhea occurs, effective symptomatic treatment should be initiated. Antiemetics should be administered to subjects according to local institutional guidelines or other guidelines (e.g., ASCO guidelines). For the treatment of febrile neutropenia, hematopoietic growth factors, namely G-CSF or GM-CSF, may be used according to institutional guidelines or other guidelines (e.g., ASCO guidelines); however, these factors are only for primary prophylaxis. For the treatment of infusion reactions related to the study drug, acetaminophen, pethidine, and chlorpheniramine or other antihistamines should be used according to institutional guidelines.

[0249] Adverse event severity assessment

[0250] The severity of adverse events was assessed using the grading scale provided in the NCI Common Terminology for Adverse Events (NCI CTCAE) v4.03 (http: / / ctep.cancer.gov / protocolDevelopment / electronic_applications / ctc.htm#ctc_40). For adverse events not listed in the NCI CTCAE, the severity was assessed using the criteria in Table 6. Heart failure was classified according to the NYHA classification criteria.

[0251] [Table 6] NCICTCAE Unspecified Adverse Event Severity Grading Scale *Instrumental ADLs refer to activities such as preparing meals, buying groceries or clothes, using the telephone, and managing money. Self-care ADLs include bathing, dressing and undressing, feeding yourself, using the toilet, taking medication, and not being bedridden.

[0252] Example 2. Clinical trial results of IMC-001 as a neoadjuvant therapy for resectable GC, ESCC, and HCC.

[0253] Security

[0254] Except for elevated G3 AST / ALT (n = 3, 6%), all treatment-related adverse events (TRAEs) were G1 or G2. TRAEs occurred in 5% or fewer patients and included hyperthyroidism (n = 9, 18%), hypothyroidism (n = 7, 14%), fatigue (n = 7, 14%), pruritus (n = 6, 12%), rash (n = 4, 8%), infusion-related reactions (n ​​= 3, 6%), and diarrhea (n = 3, 6%).

[0255] Clinical tumor response

[0256] No patients experienced disease progression during neoadjuvant therapy. According to RECIST 1.1, among the 17 patients with measurable lesions, 3 (17.6%) achieved a partial response (PR), and 14 (82.4%) had stable disease (SD). Figure 2 ).

[0257] Based on EORTC criteria, among 30 patients with evaluable metabolic response, 7 (23.3%) had a metabolic partial response (mPR), 20 (66.7%) had stable metabolic disease (mSD), and 3 (10.0%) had progressive metabolic disease (mPD). Figure 3 Of the three patients with metabolic disease progression, two showed increased tumor-infiltrating lymphocytes and tumor regression in surgical pathology.

[0258] Surgical results

[0259] All patients underwent radical R0 resection, and the pathological stage was classified as stage I (n=36, 75.0%) / stage II (n=6, 12.5%) / stage III (n=6, 12.5%). There were no surgical deaths or treatment-related surgical morbidities.

[0260] Pathological response

[0261] Although there were no major pathological responses, 51% of patients developed varying degrees of tumor necrosis or fibrosis, and 7 patients (15%) had less than 50% residual viable tumor cells. Figure 4 ).

[0262] relapse

[0263] During a median follow-up of 31.4 months (range: 1.7 to 41.2 months), only 4 patients (8.3%) experienced recurrence. Median progression-free survival (PFS) or recurrence-free survival (RFS) was not reached.

[0264] in conclusion

[0265] Neoadjuvant therapy with IMC-001 has demonstrated good tolerability and promising antitumor activity in patients with resectable microsatellite stable gastric cancer (GC), esophageal cancer (EC), and hepatocellular carcinoma (HCC).

Claims

1. A pharmaceutical composition for treating tumors or inhibiting tumor proliferation in patients with upper gastrointestinal cancer, said pharmaceutical composition comprising: A therapeutically effective amount of an anti-PD-L1 antibody, said anti-PD-L1 antibody specifically binds to PD-L1 and comprises HCDR1, HCDR2 and HCDR3 contained in the heavy chain variable region (HCVR) of SEQ ID NO:1, and LCDR1, LCDR2 and LCDR3 contained in the light chain variable region (LCVR) of SEQ ID NO:

2. in, The antibody was administered to the patient as neoadjuvant therapy prior to surgical removal of the tumor.

2. The pharmaceutical composition according to claim 1, wherein, The upper gastrointestinal cancers mentioned are stomach cancer, esophageal cancer, or liver cancer.

3. The pharmaceutical composition according to claim 1, wherein, The anti-PD-L1 antibody comprises a heavy chain variable region and a light chain variable region. The heavy chain variable region comprises HCDR1 of SEQ ID NO:5, HCDR2 of SEQ ID NO:6, and HCDR3 of SEQ ID NO:

7. The light chain variable region comprises LCDR1 of SEQ ID NO:8, LCDR2 of SEQ ID NO:9, and LCDR3 of SEQ ID NO:

10.

4. The pharmaceutical composition according to claim 3, wherein, The light chain variable region contains the amino acid sequence of SEQ ID NO:2, or contains at least 95% of the same amino acid sequence as SEQ ID NO:

2.

5. The pharmaceutical composition according to claim 4, wherein, The amino acids at positions 3 and 5 of the variable region of the light chain are the same as those at positions 3 and 5 of SEQ ID NO:

2.

6. The pharmaceutical composition according to claim 4, wherein, The heavy chain variable region contains at least 95% of the same amino acid sequence as SEQ ID NO:

1.

7. The pharmaceutical composition according to claim 1, wherein, The anti-PD-L1 antibody comprises a heavy chain having the amino acid sequence of SEQ ID NO:3 and a light chain having the amino acid sequence of SEQ ID NO:

4.

8. The pharmaceutical composition according to claim 1, wherein, The aforementioned upper gastrointestinal cancer is resectable.

9. The pharmaceutical composition according to claim 1, wherein, The aforementioned upper gastrointestinal cancer is recurrent.

10. The pharmaceutical composition according to claim 1, wherein, The upper gastrointestinal cancer is metastatic.

11. The pharmaceutical composition according to claim 1, wherein, Radical surgical treatment was performed for the aforementioned upper gastrointestinal cancer.

12. The pharmaceutical composition according to claim 2, wherein, The gastric cancer referred to is a submucosal tumor of the stomach or adenocarcinoma of the stomach (GC).

13. The pharmaceutical composition according to claim 2, wherein, The esophageal cancer mentioned is esophageal squamous cell carcinoma (ESCC).

14. The pharmaceutical composition according to claim 2, wherein, The liver cancer mentioned is hepatocellular carcinoma (HCC).

15. The pharmaceutical composition according to claim 1, wherein, The neoadjuvant therapy involves administering one or more doses of the anti-PD-L1 antibody prior to surgical resection of the tumor.

16. The pharmaceutical composition according to claim 15, wherein, Each dose is administered at intervals of 10 to 18 days.

17. The pharmaceutical composition according to claim 16, wherein, The neoadjuvant therapy involves administering the anti-PD-L1 antibody 2 to 6 times at 10 to 18-day intervals prior to surgical removal of the tumor.

18. The pharmaceutical composition according to claim 1, wherein, The neoadjuvant therapy involves administering the anti-PD-L1 antibody at a dose ranging from 10 mg / kg to 30 mg / kg.

19. The pharmaceutical composition according to claim 18, wherein, The neoadjuvant therapy involves administering the anti-PD-L1 antibody at a dose of 20 mg / kg.

20. The pharmaceutical composition according to claim 1, wherein, Surgical resection of the tumor was performed between 11 and 84 days after the last administration of the anti-PD-L1 antibody.

21. A method for treating a tumor or inhibiting tumor growth, the method comprising: (a) Select patients with upper gastrointestinal cancer; (b) Administer a therapeutically effective dose of a PD-L1 inhibitor to the patient; as well as (c) Following step (b), surgically remove the tumor of the upper gastrointestinal cancer. The PD-L1 inhibitor is an antibody that specifically binds to PD-L1. The antibody contains HCDR1, HCDR2 and HCDR3 contained in the heavy chain variable region (HCVR) of SEQ ID NO:1, and LCDR1, LCDR2 and LCDR3 contained in the light chain variable region (LCVR) of SEQ ID NO:

2.

22. A kit for treating tumors or inhibiting tumor proliferation in patients with upper gastrointestinal cancer, the kit comprising a PD-L1 inhibitor and a manual for using the PD-L1 inhibitor as neoadjuvant therapy. in, The manual contains instructions for administering the PD-L1 inhibitor intravenously 2 to 6 times at intervals of 10 to 18 days and at a dose of 20 mg / kg prior to surgical resection of the tumor.

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