Neoadjuvant therapy using PD-L1 inhibitors for cancer treatment

Abstract

The present invention relates to a method for treating a tumor or inhibiting tumor growth, comprising administering a therapeutically effective dose of a PD-L1 (programmed death-ligand 1) inhibitor (e.g., an anti-PD-L1 antibody) as neoadjuvant therapy to cancer patients in need, followed by surgical resection. In particular, the present invention relates to a method for administering a PD-L1 inhibitor as neoadjuvant therapy to patients with surgically resectable upper gastrointestinal cancers, such as surgically resectable localized gastric cancer, esophageal cancer, or liver cancer. The administration of PD-L1 inhibitors, particularly anti-PD-L1 antibodies, has been confirmed to show excellent therapeutic efficacy as neoadjuvant therapy in patients with surgically resectable localized gastric cancer, esophageal cancer, and liver cancer.

Description

Technical Field

[0001] The present invention relates to a method for treating a tumor or inhibiting tumor growth, which comprises administering a PD-L1 (programmed death-ligand 1) inhibitor (for example, anti-PD-L1 antibody IMC-001 or its biological equivalent) in a therapeutically effective amount as neoadjuvant therapy to a cancer patient, and then performing surgical resection. In particular, the present invention relates to a method for administering a PD-L1 inhibitor as preoperative adjuvant therapy to a patient with upper gastrointestinal cancer capable of surgical resection, such as local gastric cancer, esophageal cancer or liver cancer.

[0002] Sequence List This application includes a sequence listing, which is submitted electronically in XML format and is hereby incorporated by reference in its entirety. A copy of the sequence listing created on April 13, 2024 has the name KC24060.xml and a size of 12.3 kilobytes.

Background Art

[0003] As one of the main characteristics of cancer, immune evasion has recently attracted attention, and its mechanism is being clarified with the development of tumor immunology. An immune checkpoint is a concept that includes immunosuppressive regulatory molecules that prevent immune cells from attacking self-cells. As one of the various mechanisms by which tumor cells evade immune surveillance, these immune checkpoints are used to form an immunosuppressive environment within the tumor. Immunotherapy strategies that achieve anti-cancer effects by activating the suppressed anti-tumor immune system have gained attention, and representative immune checkpoint inhibitors, such as CTLA-4 inhibitors or PD-1 / PD-L1 inhibitors, have recently demonstrated improved survival rates in various cancer types, establishing them as one of the standard cancer treatments for immunotherapy and presenting a new paradigm in cancer treatment (Pardoll DM: The blockade of immune checkpoints in cancer immunotherapy. Nat Rev Cancer 12:252-64, 2012). A study on lung cancer has shown that the therapeutic effect of immune checkpoint inhibitors is higher the more mutations there are in the tumor, and especially higher when there are many clonal neoantigens. However, even if the overall number of mutations is high, if there are many subclonal neoantigens, the therapeutic effect of PD-1 inhibitors is low (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).

[0004] Furthermore, considering that the number of new subclonal antigens increases with tumor evolution as cancer progresses, and the immunosuppressive microenvironment advances, it is possible that immune checkpoint inhibitors are more effective in early-stage localized cancer than in stage IV metastatic cancer (Jamal-Hanjani M, Quezada SA, Larkin J, et al: Translational implications of tumor heterogeneity. 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). Immune checkpoint inhibitors such as pembrolizumab, nivolumab, and ipilimumab have shown improved survival rates or tumor reduction in unresectable localized advanced or metastatic gastric cancer, esophageal cancer, and hepatocellular carcinoma, and the effectiveness of immune checkpoint inhibitor treatment strategies in these cancer types has already been demonstrated. On the other hand, surgical resection is the treatment option for surgically treatable gastric cancer, esophageal cancer, and hepatocellular carcinoma. However, tumor recurrence after surgery is common, and early recurrence is also observed. Therefore, there is a pressing need for safe and effective treatment methods for surgically treatable cancers. [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] Pardoll DM, The blockade of immune checkpoints in cancer immunotherapy., Nat Rev Cancer, 2012, Vol. 12, p.252-264 [Non-Patent Document 2] McGranahan N, Furness AJ, Rosenthal R, et al, Clonal neoantigens elicit T cell immunoreactivity and sensitivity to immune checkpoint blockade., Science, 2016, Vol. 351, p.1463-1469 [Non-Patent Document 3] Jamal-Hanjani M, Quezada SA, Larkin J, et al, Translational implications of tumor heterogeneity., Clin Cancer Res, 2015, Vol. 21, p.1258-1266 [Non-Patent Document 4] Gil Del Alcazar CR, Huh SJ, Ekram MB, et al, Immune Escape in Breast Cancer During In Situ to Invasive Carcinoma Transition., Cancer Discov, 2017, Vol. 7, p.1098-1115 [Overview of the Initiative] [Problems that the invention aims to solve]

[0006] The purpose of this invention is to solve all of the problems of the prior art described above. One objective of the present invention is to provide preoperative therapy, cancer treatment methods, pharmaceutical compositions, kits, and administration methods for patients with surgically resectable upper gastrointestinal cancers. The objectives of the present invention are not limited to those stated above. The objectives of the present invention will become clearer from the following description and will be implemented by the means and combinations thereof described in the claims. [Means for solving the problem]

[0007] A typical configuration of the present invention for achieving the above objective is as follows. One aspect of the present invention provides a pharmaceutical composition for treating tumors in patients with upper gastrointestinal cancer or inhibiting tumor growth, comprising a therapeutically effective amount of a PD-L1 inhibitor (for example, an anti-PD-L1 antibody that specifically binds to PD-L1 and includes 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), which is administered to the patient as neoadjuvant therapy before surgical resection of the tumor. Another aspect of the present invention provides a method for treating a tumor or inhibiting tumor growth, comprising: (a) screening patients for upper gastrointestinal cancer; (b) administering a therapeutically effective dose of a PD-L1 inhibitor (for example, an anti-PD-L1 antibody that specifically binds to PD-L1 and includes 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 resecting an upper gastrointestinal cancer tumor after step (b).

[0008] In one embodiment, the upper gastrointestinal cancer may be gastric cancer, esophageal cancer, or liver cancer. In one embodiment, the anti-PD-L1 antibody may include a heavy chain variable region comprising HCDR1 of SEQ ID NO: 5, HCDR2 of SEQ ID NO: 6, and HCDR3 of SEQ ID NO: 7, and a light chain variable region comprising LCDR1 of SEQ ID NO: 8, LCDR2 of SEQ ID NO: 9, and LCDR3 of SEQ ID NO: 10. In one embodiment, the light chain variable region of the anti-PD-L1 antibody may contain or be composed of the amino acid sequence of SEQ ID NO: 2, or an amino acid sequence that is 95% or more identical to SEQ ID NO: 2. In one embodiment, the heavy chain variable region of the anti-PD-L1 antibody may contain or be composed of the amino acid sequence of SEQ ID NO: 1, or an amino acid sequence that is 95% or more identical to SEQ ID NO: 1.

[0009] In one embodiment, the anti-PD-L1 antibody may include 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. In one embodiment, the anti-PD-L1 antibody may be a fully human antibody. In one embodiment, the anti-PD-L1 antibody may retain an Fc effector function that can stimulate antibody-dependent cellular cytotoxicity (ADCC). In one embodiment, the anti-PD-L1 antibody may be a fully human antibody of the IgG class. In one embodiment, the anti-PD-L1 antibody may be a fully human antibody of the IgG1 or IgG4 class.

[0010] In one embodiment, an anti-PD-L1 antibody may be administered intravenously or subcutaneously as neoadjuvant therapy. In one embodiment, the upper gastrointestinal cancer may be resectable. In one embodiment, the upper gastrointestinal cancer may be recurrent. In one embodiment, the upper gastrointestinal cancer may be metastatic. In one embodiment, the goal of surgery for upper gastrointestinal cancer may be curative. In one embodiment, gastric cancer may be a gastric submucosal tumor or a gastric adenocarcinoma (GC). In one embodiment, the esophageal cancer may be esophageal squamous cell carcinoma (ESCC). In one embodiment, the liver cancer may be hepatocellular carcinoma (HCC).

[0011] In one embodiment, as neoadjuvant therapy, the anti-PD-L1 antibody may be administered once or more times, with each dose administered at intervals of once a week, once every two weeks, once every three weeks, once every four weeks, once every five weeks, or once every six weeks. In one embodiment, as neoadjuvant therapy, the anti-PD-L1 antibody may be administered one or more times, and each dose may be administered at intervals of once every two weeks. In one embodiment, as neoadjuvant therapy, the anti-PD-L1 antibody may be administered once, twice, three times, four times, five times, or six times before surgical resection of the tumor. In one embodiment, as neoadjuvant therapy, the anti-PD-L1 antibody may be administered at a dose of 10 mg / kg to 30 mg / kg based on the patient's body weight. In one embodiment, as neoadjuvant therapy, the anti-PD-L1 antibody may be administered at a dose of 20 mg / kg. In one embodiment, surgical resection of the tumor may be performed between 11 days and 84 days or between 11 days and 42 days after the last administration of the anti-PD-L1 antibody.

[0012] Another aspect of the present invention provides a kit containing a PD-L1 inhibitor (for example, an anti-PD-L1 antibody that specifically binds to PD-L1 and contains HCDR1, HCDR2, and HCDR3 included in the heavy chain variable region (HCVR) of SEQ ID NO: 1 and LCDR1, LCDR2, and LCDR3 included in the light chain variable region (LCVR) of SEQ ID NO: 2) together with instructions for use as neoadjuvant therapy for treating tumors or inhibiting tumor growth in upper gastrointestinal cancer patients. In one embodiment, the instructions for use included in the kit include administering a PD-L1 inhibitor (for example, an anti-PD-L1 antibody that specifically binds to PD-L1 and contains HCDR1, HCDR2, and HCDR3 included in the heavy chain variable region (HCVR) of SEQ ID NO: 1 and LCDR1, LCDR2, and LCDR3 included in the light chain variable region (LCVR) of SEQ ID NO: 2) to upper gastrointestinal cancer patients at a dose of 20 mg / kg at intervals of 10 days to 18 days once, twice, three times, four times, five times, or six times before surgical resection of the tumor. Another aspect of the present invention provides a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody that specifically binds to PD-L1 and includes HCDR1, HCDR2, and HCDR3 included in the heavy chain variable region (HCVR) of SEQ ID NO: 1 and LCDR1, LCDR2, and LCDR3 included in the light chain variable region (LCVR) of SEQ ID NO: 2) for use in a method of treating a tumor or inhibiting tumor growth, the method comprising: (a) screening a patient with upper gastrointestinal cancer; (b) administering to the patient a therapeutically effective amount of a PD-L1 inhibitor; and (c) surgically excising the upper gastrointestinal cancer tumor after step (b).

[0013] Another aspect of the present invention provides a method of treating a tumor or inhibiting tumor growth, the method comprising: (a) screening 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 and includes HCDR1, HCDR2, and HCDR3 included in the heavy chain variable region (HCVR) of SEQ ID NO: 1 and LCDR1, LCDR2, and LCDR3 included in the light chain variable region (LCVR) of SEQ ID NO: 2); and (c) surgically excising the gastric cancer tumor after step (b). Another aspect of the present invention provides a method of treating a tumor or inhibiting tumor growth, the method comprising: (a) screening 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 and includes HCDR1, HCDR2, and HCDR3 included in the heavy chain variable region (HCVR) of SEQ ID NO: 1 and LCDR1, LCDR2, and LCDR3 included in the light chain variable region (LCVR) of SEQ ID NO: 2); and (c) surgically excising the esophageal cancer tumor after step (b). Another aspect of the present invention provides a method for treating a tumor or inhibiting tumor growth, comprising the steps of (a) screening patients for resectable liver cancer; (b) administering a therapeutically effective dose of a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody that specifically binds to PD-L1 and includes 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 resecting the liver cancer tumor after step (b). [Effects of the Invention]

[0014] Preoperative adjuvant therapy using anti-PD-L1 antibodies showed good tolerability and promising antitumor activity in patients with microsatellite-stable, resectable gastric, esophageal, and hepatocellular carcinomas. In one embodiment of the present invention, administration of IMC-001 as preoperative pre-treatment resulted in low-grade treatment-related adverse events (TRAEs) and measurable lesions, with partial remission (PR) or stable disease (SD). In patients where metabolic response could be evaluated, metabolic partial remission (mPR), metabolic stable disease (mSD), or metabolic progression (mPD) were observed. Furthermore, administration of IMC-001 as preoperative pre-treatment resulted in varying levels of tumor necrosis or fibrosis, and in some patients, a reduction in residual surviving tumor cells to less than 50% was confirmed. [Brief explanation of the drawing]

[0015] [Figure 1] This figure schematically illustrates the procedure for a clinical trial according to one embodiment of the present invention. [Figure 2] This figure shows the results of a clinical trial according to one embodiment of the present invention, illustrating the clinical tumor response. PR indicates partial remission, and SD indicates stable disease. [Figure 3]This figure shows the results of a clinical trial according to one embodiment of the present invention, illustrating the clinical tumor response. mPR indicates metabolic partial response, mSD indicates metabolic stable disease, and mPD indicates metabolic progressive disease. [Figure 4] This figure shows the results of a clinical trial according to one embodiment of the present invention, specifically a diagram illustrating a pathological reaction. [Modes for carrying out the invention]

[0016] The detailed description below of the present invention will be given with reference to specific embodiments in which the invention can be implemented (if drawings exist), but the present invention is not limited thereto and is limited only to the appended claims in all scope identical or equivalent to that described herein. It should be understood that the various embodiments / examples of the present invention are different from each other but do not need to be mutually exclusive. For example, specific shapes, structures and characteristics described herein may be changed from one embodiment / example to another, or realized by combining multiple embodiments / examples, as long as they do not depart from the technical idea and scope of the present invention. Technical and scientific terms used herein have their ordinary meanings in the art unless otherwise defined. In case of any conflict, the description herein, including the definitions, shall prevail. Furthermore, the definitions of terms given herein are for the purpose of interpretation herein, and a singular term shall be interpreted to include the plural form (i.e., at least one) unless otherwise inappropriate in the context, and vice versa.

[0017] Cancer treatment or cancer growth inhibition method One aspect of the present invention provides a method for treating a tumor or inhibiting tumor growth, comprising the steps of screening patients for 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 before surgical resection of the tumor in the patient. In one embodiment, the method disclosed herein may further include the step of administering a PD-L1 inhibitor (e.g., anti-PD-L1 antibody IMC-001 or its bioequivalent) to a patient as adjuvant therapy after completing surgery to treat upper gastrointestinal cancer (e.g., gastric cancer, esophageal cancer, or liver cancer).

[0018] As used in this application, "gastric cancer" is a general term for carcinomas (malignant tumors) that occur in the stomach, and examples of malignant tumors that occur in the stomach include gastric adenocarcinoma, lymphoma, gastric submucosal tumor, and leiomyosarcoma. Gastric cancer can occur in any part of the stomach and can metastasize to the entire stomach and other organs, particularly the esophagus, lungs, and liver. In one embodiment, gastric cancer is gastric adenocarcinoma. In one embodiment, gastric cancer is resectable and recurrent. In one embodiment, gastric cancer is metastatic. As used in this application, "esophageal cancer" is cancer that occurs in the esophagus and is classified by location into cervical esophageal cancer, thoracic esophageal cancer, and gastroesophageal junction cancer, and by cell morphology into squamous cell carcinoma, adenocarcinoma, sarcoma, lymphoma, melanoma, etc. In one embodiment, esophageal cancer is esophageal squamous cell carcinoma (ESCC). In one embodiment, esophageal cancer is resectable and recurrent. In one embodiment, esophageal cancer is metastatic. As used in this application, “liver cancer” refers to cancers of the liver, such as hepatocellular carcinoma, lamellar carcinoma, cholangiocarcinoma, angiosarcoma, and hepatoblastoma. In one embodiment, liver cancer is hepatocellular carcinoma (HCC). In one embodiment, liver cancer is resectable and recurrent. In one embodiment, liver cancer is metastatic. In some embodiments, the patient is a candidate for surgery to remove a tumor of gastric cancer, esophageal cancer, or liver cancer. In some embodiments, the patient has gastric cancer, esophageal cancer, or liver cancer that is a candidate for surgery aimed at curative treatment.

[0019] As used in this application, the terms “to treat,” “to cure,” or similar expressions mean the alleviation or reduction of the severity of at least one symptom or sign, the temporary or permanent elimination of a symptomatic factor, the delay or inhibition of tumor growth, the reduction of tumor cell load or tumor burden, the promotion of tumor regression, the reduction of tumor size, the induction of necrosis and / or disappearance, the prevention of tumor recurrence, the prevention or inhibition of metastasis, the inhibition of metastatic tumor growth, the elimination of the need for surgery, and / or the extension of the subject’s survival. In some embodiments, the terms “tumor,” “lesion,” “tumor lesion,” “cancer,” and “malignant” are used interchangeably and mean one or more malignant growths. As used in this application, the term “recurrence” means frequent or recurrent diagnoses of gastric, esophageal, or liver cancer in a patient, or recurrence of a primary tumor and / or previous tumor, and may also mean the frequent or recurrent occurrence of individual tumors, such as new tumors showing recurrence of a primary tumor or previous tumor. In certain embodiments, administration of a PD-L1 inhibitor inhibits the recurrence of gastric, esophageal, or liver cancer in a patient.

[0020] As used in this application, “Subject requiring a PD-L1 inhibitor” means a human or non-human mammal exhibiting one or more symptoms or signs of gastric cancer, esophageal cancer, or liver cancer, or being diagnosed with gastric cancer, esophageal cancer, or liver cancer and requiring treatment for it. In many embodiments, the terms “subject” and “patient” are used interchangeably. Such expressions include subjects with primary, established, or recurrent tumors (advanced malignancies). In certain embodiments, this expression includes human subjects who have been treated for and / or require treatment for recurrent but non-metastatic gastric, esophageal, or liver cancer. In certain embodiments, this expression includes patients with solid tumors that have not been adequately controlled by previous therapies (e.g., surgery or treatment with other anticancer agents other than IMC-001 or its bioequivalents), or who are resistant or refractory. In certain embodiments, this expression includes subjects with gastric, esophageal, or liver cancer that are candidates for curative surgery.

[0021] In certain embodiments, the methods of the present invention are used to treat subjects having solid tumors. As used in this application, the term “solid tumor” typically refers to an abnormal mass of tissue that does not contain cysts or fluid areas. Solid tumors can be benign (not cancerous) or malignant (cancerous). For the purposes of this invention, the term “solid tumor” means malignant solid tumor. This term includes various types of solid tumors named according to the type of cells that form them, namely sarcomas, carcinomas, and progenitor tumors.

[0022] In certain embodiments, the method of the present invention involves administering a therapeutically effective dose of a PD-L1 inhibitor (e.g., anti-PD-L1 antibody IMC-001 or its bioequivalent) in combination with an additional therapeutic substance or therapy. The additional therapeutic substance or therapy may be applied to enhance the antitumor effect, to reduce the toxic effects of one or more therapies, and / or to reduce the dose of one therapy. In various embodiments, the additional therapeutic substance or therapy may include one or more of the following: antiviral therapy (e.g., cidofovir), photodynamic therapy, PD-1 inhibitor (e.g., nivolumab, pembrolizumab, etc., antibodies against PD-1 well known to those skilled in the art), lymphocyte-activating gene 3 (LAG3) inhibitor (e.g., anti-LAG3 antibody), cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) inhibitor (e.g., ipilimumab (ip)). ilimumab), glucocorticoid-induced tumor necrosis factor receptor (GITR) agonists (e.g., anti-GITR antibodies), T cell immunoglobulin and mucin-containing molecule 3 (TIM3) inhibitors, B and T lymphocyte attenuation factor (BTLA) inhibitors, T cell immune receptor (TIGIT) inhibitors with Ig and ITIM domains, CD38 inhibitors, CD47 inhibitors, other T cell co-suppressive molecules, or ligand antagonists (e.g., CD28, 2B4, LY) Antibodies against 108, 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., "VEGFTrap" such as aflibercept, or other VEGF inhibitors as described in US7087411) Fusion proteins, or anti-VEGF antibodies, or their antigen-binding fragments (e.g., bevacizumab or ranibizumab), small molecule kinase inhibitors of the VEGF receptor (e.g., sunitinib, sorafenib, pazopanib, or ramucirumab), angiopoietin-2 (Ang2) inhibitors,Transforming growth factor beta (TGFβ) inhibitors, epidermal growth factor receptor (EGFR) inhibitors (e.g., erlotinib, cetuximab), agonists against costimulatory receptors (e.g., agonists against CD28, 4-1BB, or OX40), antibodies against tumor-specific antigens (e.g., CA9, CA125, melanoma-associated antigen 3 (MAGE3), carcinoembryonic antigen (CEA), vimentin, tumor M2-PK, prostate-specific antigen (PSA), whip (e.g., N-1, MART-1 and CA19-9), vaccines (e.g., Bacillus Calmet-Guérin 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) (doxorubicin), daunorubicin, cisplatin, carboplatin, gemcitabine, methotrexate, mitoxantrone, oxaliplatin, paclitaxel, topotecan, irinotecan, vinorelbine lbine 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 such as 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 agents such as corticosteroids,Nutritional supplements such as non-steroidal anti-inflammatory drugs (NSAIDs) and antioxidants.

[0023] As used in this application, the term “antiviral therapy” refers to any substance, drug, or therapy used on a host for the treatment, prevention, or improvement of a viral infection, including but not limited to zidovudine, lamivudine, abacavir, ribavirin, lopinavir, efavirenz, cobicistat, tenofovir, rilpivirine, analgesics, corticosteroids, and combinations thereof. In the context of this invention, 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). In certain embodiments, administering a therapeutically effective dose of a PD-L1 inhibitor (e.g., anti-PD-L1 antibody IMC-001 or its bioequivalent) to subjects with gastric, esophageal, or liver cancer results in increased inhibition of tumor growth, for example, leading to greater tumor regression in the treated subjects.

[0024] In certain embodiments, administration of a PD-L1 inhibitor achieves one or more of the following: (i) improved tumor response rates, e.g., improved overall response rates, complete remissions, or partial remissions compared to subjects treated with surgical resection alone or untreated subjects; (ii) delayed tumor growth and development, e.g., delayed tumor growth by approximately 3 days, >3 days, approximately 7 days, >7 days, >15 days, >1 month, >3 months, >6 months, >1 year, >2 years, or >3 years compared to subjects treated with surgical resection alone or untreated subjects; (iii) increased disease-free survival (DFS) to tumor recurrence or death compared to subjects treated with surgical resection alone or untreated subjects; and (iv) improved overall response rates, complete remissions, or partial remissions compared to subjects treated with surgical resection alone or untreated subjects. In certain embodiments, administration of a therapeutically effective dose of a PD-L1 inhibitor (e.g., anti-PD-L1 antibody IMC-001, or its bioequivalent) to subjects with gastric, esophageal, or liver cancer prevents tumor recurrence and / or extends the subject's survival, for example, by >15 days, >1 month, >3 months, >6 months, >12 months, >18 months, >24 months, >36 months, or >48 months compared to subjects treated with surgical resection alone or untreated subjects.

[0025] In certain embodiments, administration of a therapeutically effective dose of a PD-L1 inhibitor (e.g., anti-PD-L1 antibody IMC-001, or its bioequivalent) to subjects with gastric, esophageal, or liver cancer results in an increase in overall survival (OS) or progression-free survival (PFS) compared to subjects treated with surgical resection alone. In certain embodiments, PFS is increased by 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 certain embodiments, overall survival (OS) is increased by 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 individuals treated with surgical resection alone.

[0026] PD-L1 inhibitors The methods disclosed herein include administering a PD-L1 inhibitor in a therapeutically effective dose, wherein the PD-L1 inhibitor may be an anti-PD-L1 antibody, for example, a fully human anti-PD-L1 antibody IMC-001 or its bioequivalent. As used herein, the term “bioequivalent” means an anti-PD-L1 antibody or PD-L1 binding protein, or a fragment thereof, which, when administered in the same molar dose in one or more doses under similar experimental conditions, exhibits no significant difference in absorption rate and / or absorption level compared to the IMC-001 antibody. In the context of the present invention, the term “bioequivalent” includes an antibody-binding protein that binds to PD-L1 and is clinically indistinguishable from the IMC-001 antibody in terms of safety, purity and / or efficacy.

[0027] As used in this application, 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) interconnected by disulfide bonds, as well as its polymer (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 invariant 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 invariant region (CL). The VH and VL regions can be further subdivided into highly variable regions called complementarity-determining regions (CDRs), with more conserved regions called framework regions (FRs) distributed between them. Each VH and VL consists of three CDRs and four FRs, arranged in the following order from the amino-terminus to the carboxy-terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In certain embodiments, the FR of an antibody (or its antigen-binding fragment) may be identical to a human germline sequence and may be naturally or artificially modified. The amino acid consensus sequence can be defined based on side-by-side analysis of two or more CDRs. As used in this application, the term “antibody” also includes antigen-binding fragments of full-length antibody molecules.

[0028] As used in this application, terms such as “antigen-binding fragment” and “antigen-binding moiety” of an antibody include naturally occurring, enzymatically obtained, or synthetically or genetically engineered polypeptides or glycoproteins that specifically bind to an antigen to form a complex. Antigen-binding fragments of antibodies can be derived from full-length antibody molecules using any suitable standard technique, such as protein hydrolysis or recombinant genetic engineering techniques involving the manipulation and expression of DNA encoding the antibody’s variable and selectively invariant domains. Such DNA is readily available from known and / or commercially available sources, such as DNA libraries (e.g., phage antibody libraries), or can be synthesized. DNA can be sequenced and manipulated using chemical or molecular biological techniques, for example, by arranging one or more variable and / or invariant domains in a suitable configuration, introducing codons, constructing cysteine ​​residues, or modifying, adding, or removing amino acids.

[0029] Non-restrictive examples of antigen-binding fragments include: (i) Fab fragments, (ii) F(ab')2 fragments, (iii) Fd fragments, (iv) Fv fragments, (v) short-chain Fv(scFv) molecules, (vi) dAb fragments, (vii) minimal recognition units consisting of amino acid residues that mimic the highly variable region of an antibody (e.g., isolated complementarity-determining regions (CDRs) such as the CDR3 peptide), or restricted FR3-CDR3-FR4 peptides. Domain-specific antibodies, single-domain antibodies, domain-deficient antibodies, chimeric antibodies, CDR-grafted antibodies, diabodies, triabodies, tetrabodies, minibodies, nanobodies (e.g., monovalent nanobodies, bivalent nanobodies, etc.), small modular immunopharmaceuticals (SMIPs), and other manipulated molecules such as shark variable IgNAR domains are also included in the expression "antigen-binding fragment" as used in this application.

[0030] Antibody antigen-binding fragments typically contain at least one variable domain. The variable domain may be of any size or amino acid composition and typically contains at least one CDR adjacent to or within the same frame as one or more framework sequences. In antigen-binding fragments having a VH domain conjugated to a VL domain, the VH and VL domains can be arranged relative to each other in any suitable configuration. For example, the variable region may be a dimer and may contain a VH-VH, VH-VL, or VL-VL dimer. Alternatively, the antibody antigen-binding fragment may contain a monomeric VH or VL domain.

[0031] In certain embodiments, the antigen-binding fragment of the antibody may include at least one invariant domain and at least one variable domain covalently linked thereto. Non-limiting exemplary structures of variable and invariant domains that may be found in the antigen-binding fragment of the antibody of the present invention 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 arrangement of variable and immutable domains including any of the exemplary structures described above, the variable and immutable domains may be directly linked to each other, or they may be linked to each other by the entire hinge, a portion thereof, or a linker region. The hinge region may consist of at least two (e.g., five, ten, fifteen, twenty, forty, sixty, or more) amino acids that form a single polypeptide molecule through flexible or semi-flexible linkages between adjacent variable and / or immutable domains. Furthermore, the antigen-binding fragment of the antibody of the present invention may include, non-covalently (e.g., by disulfide bonds) homodimers or heterodimers (or other multimers) of any of the aforementioned variable and immutable domain sequences, linked to each other and / or one or more monomeric VH or VL domains.

[0032] The antibody used in the method of the present invention may be a human antibody. As used in this application, the term “human antibody” refers to an antibody having variable domains and invariant regions derived from a human germline immunoglobulin sequence. Nevertheless, the human antibody of the present invention may include amino acid residues not encoded by the human germline immunoglobulin sequence (e.g., mutations introduced by random or site-directed mutagenesis in vitro or by somatic mutation in vivo), for example, in the CDR, particularly CDR3. However, as used in this application, the term “human antibody” is not intended to include antibodies in which a CDR sequence derived from the germline of another mammalian species, such as mouse, is grafted onto a human framework sequence. In one embodiment, all variable and invariant domains may be derived from a human immunoglobulin sequence (a fully human antibody).

[0033] The antibodies used in the methods disclosed herein may be recombinant human antibodies. As used herein, the term “recombinant human antibody” encompasses all human antibodies produced, expressed, constructed or isolated by recombinant means, including antibodies expressed using recombinant expression vectors transfected into host cells (as further described below), antibodies isolated from recombinant or combined human antibody libraries (as further described below), antibodies isolated from animals transgenic to human immunoglobulin genes (e.g., mice) (see, for example, literature [Taylor et al. (1992) Nucl. Acids Res. 20:6287-6295]), or antibodies produced, expressed, constructed or isolated by any other means via splicing of human immunoglobulin gene sequences into DNA sequences. These recombinant human antibodies have variable and invariant regions derived from human germline immunoglobulin sequences. However, in certain embodiments, these recombinant human antibodies are mutagenic in vitro (or, if transgenic animals for the human Ig sequence are used, somatically mutagenic in vivo), and therefore the amino acid sequences of the VH and VL regions of the recombinant antibodies are derived from and related to human germline VH and VL sequences, but are sequences that may not naturally exist in the in vivo human antibody germline repertoire.

[0034] 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 composed 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 composed 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), where 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 certain embodiments, the anti-PD-L1 antibody comprises an HCVR comprising or being composed of SEQ ID NO: 1 and an LCVR comprising or being composed of SEQ ID NO: 2. In certain embodiments, the anti-PD-L1 antibody comprises a heavy chain and a light chain, the heavy chain comprising or comprising the amino acid sequence of SEQ ID NO: 3, and the light chain comprising or comprising 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".

[0035] In some embodiments, the anti-PD-L1 antibody may include an HCVR having 90% or more, 95% or more, 97% or more, or 98% or more sequence identity with SEQ ID NO: 1. In some embodiments, the anti-PD-L1 antibody may include an LCVR having 90% or more, 95% or more, 97% or more, or 98% or more sequence identity with SEQ ID NO: 2. In some embodiments, the anti-PD-L1 antibody may include an HCVR having 90% or more, 95% or more, 97% or more, or 98% or more sequence identity with SEQ ID NO: 1, and an LCVR having 90% or more, 95% or more, 97% or more, or 98% or more sequence identity with SEQ ID NO: 2. Sequence identity can be measured by methods known in the art (e.g., GAP, BESTFIT, and BLAST).

[0036] In some embodiments, sequence identity may be for regions excluding the CDR region (e.g., framework sequences). In some embodiments, the bioequivalent of IMC-001 antibody is an anti-PD-L1 antibody containing HCVR having 90% or more, 95% or more, 97% or more, or 98% or more sequence identity with respect to regions excluding the HCDR1, HCDR2, and HCDR3 regions in SEQ ID NO: 1. In some embodiments, the bioequivalent of IMC-001 antibody is an anti-PD-L1 antibody containing LCVR having 90% or more, 95% or more, 97% or more, or 98% or more sequence identity with respect to regions excluding the LCDR1, LCDR2, and LCDR3 regions in SEQ ID NO: 2. In some embodiments, the bioequivalent of IMC-001 is an anti-PD-L1 antibody containing HCVR having 90% or more, 95% or more, 97% or more, or 98% or more sequence identity with respect to the region of SEQ ID NO: 1 excluding the HCDR1, HCDR2, and HCDR3 regions, and LCVR having 90% or more, 95% or more, 97% or more, or 98% or more sequence identity with respect to the region of SEQ ID NO: 2 excluding the LCDR1, LCDR2, and LCDR3 regions. In some embodiments, the bioequivalent of the IMC-001 antibody is an anti-PD-L1 antibody containing LCVR having 90% or more, 95% or more, 97% or more, or 98% or more sequence identity with respect to the region of SEQ ID NO: 2 excluding the LCDR1, LCDR2, and LCDR3 regions, and based on SEQ ID NO: 2, the 3rd and 5th amino acids from the N-terminus to the C-terminus are identical to the 3rd and 5th amino acids of SEQ ID NO: 2.

[0037] In some embodiments, the IMC-001 antibody or its bioequivalent can be produced, particularly in CHO cells, without light chain fragmentation, by having the aforementioned light chain amino acid sequence. In some embodiments, the bioequivalent of IMC-001 antibody is an anti-PD-L1 antibody comprising HCVR having the amino acid sequence of SEQ ID NO: 1 and having 5 or fewer amino acid substitutions. In some embodiments, the bioequivalent of IMC-001 is an anti-PD-L1 antibody comprising LCVR having the amino acid sequence of SEQ ID NO: 2 and having 2 or fewer amino acid substitutions. In some embodiments, the bioequivalent of IMC-001 is an anti-PD-L1 antibody comprising HCVR having the amino acid sequence of SEQ ID NO: 1 and having 5 or fewer amino acid substitutions, and LCVR having the amino acid sequence of SEQ ID NO: 2 and having 2 or fewer amino acid substitutions.

[0038] The antibodies used in the methods disclosed herein may be humanized antibodies. As used herein, the term “humanized antibody” means that, by means of one or more amino acid substitutions, deletions, and / or additions, it has a sequence different from that of an antibody of a non-human species, and that, when administered to a human subject, is less likely to induce an immune response and / or induces a milder immune response than an antibody of a non-human species. In one embodiment, a humanized antibody is produced by mutating specific amino acids in the framework and immutable domains of the heavy and / or light chains of a non-human species antibody. In another embodiment, an immutable domain of a human antibody is fused to a variable domain of a non-human species. In another embodiment, the potential immunogenicity of a non-human antibody when administered to a human subject can be reduced by altering one or more amino acid residues of one or more CDR sequences of a non-human antibody, wherein the altered amino acid residues are not important for the antibody’s immune-specific binding to the antigen, or the alteration of the amino acid sequence is a conservative alteration, and the binding of the humanized antibody to the antigen is not significantly worse than the binding of the non-human antibody to the antigen. Examples of methods for producing humanized antibodies are described in U.S. Patents 6,054,297, 5,886,152, and 5,877,293.

[0039] In one embodiment, the anti-PD-L1 antibody is applied to the PD-L1 epitope at a rate of 10 -6This is a fully human antibody of the IgG class that binds with an affinity of M or less. In one embodiment, the anti-PD-L1 antibody may be IgG1 or IgG4. In one embodiment, a fully human antibody of the IgG class (e.g., IgG1 or IgG4) that binds to the PD-L1 epitope is provided, the antibody comprising a heavy chain variable domain containing an amino acid sequence at least 90% identical to the amino acid sequence described in SEQ ID NO: 1, and a light chain variable domain containing an amino acid sequence at least 90% identical to the amino acid sequence described in SEQ ID NO: 2. In one embodiment, the present invention provides a fully human antibody of the IgG class (e.g., IgG1 or IgG4) that binds to the PD-L1 epitope, the antibody comprising a heavy chain variable domain containing a CDR1 domain, a CDR2 domain, and a CDR3 domain described in the amino acid sequences of SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7, respectively, and a light chain variable domain containing a CDR1 domain, a CDR2 domain, and a CDR3 domain described in the amino acid sequences of SEQ ID NO: 8, SEQ ID NO: 9, and SEQ ID NO: 10, respectively. More specific details regarding the anti-PD-L1 antibody of the present invention can be found in International Publication WO 2017 / 132562 A1. The IMC-001 antibody includes the sequence of the heavy chain variable domain region of SEQ ID NO: 1 and the sequence of the light chain variable domain region of SEQ ID NO: 2 of the aforementioned document, and all disclosures of the aforementioned document are incorporated herein by reference.

[0040] Manufacturing of PD-L1 inhibitors Antigen-binding proteins can be produced by any number of known techniques. In one embodiment, the present invention provides a monoclonal antibody that binds to PD-L1. Monoclonal antibodies can be purified, for example, from cells that naturally express them (for example, antibodies can be purified from hybridomas that produce them), or they can be generated in a recombinant expression system using any technique known in the art. See, for example, the literature [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)]. Monoclonal antibodies can be produced using any technique known to the art, for example, by immortalizing spleen cells isolated from transgenic animals after completion of an immunization schedule. These spleen cells can be immortalized using any technique known to the art, for example, by fusing them with myeloma cells to generate hybridomas. Myeloma cells for use in the hybridoma generation fusion procedure are preferably non-antibody-producing myeloma cells that exhibit high fusion efficiency and have enzyme deficiencies that prevent their growth in specific selective media that support the growth of only preferred fusion cells (hybridoms). Examples of cell lines suitable for use in 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-GTG 1.7, and S194 / 5XX0 Bul. Examples of cell lines used in rat fusion include R210.RCY3, Y3-Ag 1.2.3, IR983F, and 48210. Other cell lines useful for cell fusion include U-266, GM1500-GRG2, LICR-LON-HMy2, and UC729-6. In one example, a polypeptide is produced by a recombinant DNA method comprising the steps of 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.

[0041] For recombinant production of anti-PD-L1 antibodies, the nucleic acids encoding the antibodies are isolated and inserted into one or more vectors for further cloning and / or expression in host cells. These nucleic acids can be readily isolated and sequenced using known procedures (e.g., the use of oligonucleotide probes that can specifically bind to the genes encoding the heavy and light chains of the antibody). The nucleic acids encoding the anti-PD-L1 antibody (or fragment) disclosed herein can be chemically synthesized. Codon usage can be selected to improve expression in cells. Such codon usage is dependent on the selected cell type. Specific codon usage patterns have been developed not only for E. coli and other bacteria, but also for mammalian cells, plant cells, yeast cells, and insect cells. For example, see the following references: [Mayfield et al, Proc. Natl. Acad. Sci. USA. 2003 100(2):438-42], [Sinclair et al. Protein Expr. Purif. 2002 (1):96-105], [Connell N D. Curr. Opin. Biotechnol. 2001 12(5):446-9], [Makrides et al. Microbiol. Rev. 1996 60(3):512-38], [Sharp et al. Yeast. 1991 7(7):657-78].

[0042] General techniques for nucleic acid manipulation are described, for example, in the literature referenced herein by reference [Sambrook et al: 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. The polypeptide-encoding DNA is operably ligated to appropriate transcriptional or translational regulatory elements derived from mammalian, viral, or insect genes. These regulatory elements include transcriptional promoters, optional operator sequences for regulating transcription, sequences encoding appropriate mRNA-ribosome binding sites, and sequences regulating the termination of transcription and translation. They typically further include select genes that facilitate the ability to replicate in the host and recognize the transformant, which are usually conferred by the replication origin. Furthermore, recombinant DNA may contain any type of protein tag sequence useful for protein purification. 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 use with bacterial, fungal, yeast, and mammalian cell hosts are described in the literature [Cloning Vectors: A Laboratory Manual, (Elsevier, NY, 1985)].

[0043] The expression components are introduced into host cells using appropriate methods. Various methods for introducing nucleic acids into host cells are known in the art, including, but are not limited to, electroporation, transfection with calcium chloride, rubidium chloride, calcium phosphate, DEAE-dextran, and other substances, microprojectile bombardment, lipofection, and infection if the vector is an infectious formulation. Suitable host cells include prokaryotes, yeasts, mammalian cells, or bacterial cells, as will be described in more detail below.

[0044] The recombinant polypeptides (e.g., recombinant antibodies) of the present invention can be produced using any expression system known in the art. Generally, host cells are transformed using a recombinant expression vector containing DNA encoding the polypeptide of interest. Suitable host cells include prokaryotes, yeasts, or higher eukaryotic cells. Prokaryotes include Gram-negative or Gram-positive organisms, e.g., E. coli or Bacillus (bacilli). Higher eukaryotic cells include insect cells and established cell lines derived from mammals. Suitable host cells for cloning or expression of antibody-encoding vectors include the prokaryotes or eukaryotic cells described herein. For example, antibodies can be produced in bacteria, particularly when glycation and Fc effector functions are not required. For the expression of antibody fragments and polypeptides in bacteria, see, for example, U.S. Patents 5,648,237, 5,789,199, and 5,840,523. (See also the literature describing the expression of antibody fragments in E. coli [Charlton, Methods in Molecular Biology, Vol. 248 BKC Lo, ed., Humana Press, Totowa, NJ, (2003), pp.245-254]). After expression, the antibody can be isolated from the bacterial cell paste in the soluble fraction and further purified. More advanced eukaryotic cells include insect cells and established mammalian cell lines. Examples of suitable mammalian host cell lines include the COS-7 cell line of monkey kidney cells (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 (ATCC CCL 70) derived from the African green monkey kidney cell line CV1, as described in the literature [McMahan et al., 1991, EMBO J. 10: 2821].Suitable cloning and expression vectors for use with bacteria, fungi, yeast, and mammalian cell hosts have been described by Pouwels et al. [Cloning Vectors: A Laboratory Manual, (Elsevier, NY, 1985)].

[0045] In certain embodiments, vertebrate cells can be used as hosts to express anti-PD-L1 antibodies or fragments thereof. For example, mammalian cell lines adapted for growth in suspension may be useful. Other examples of useful mammalian host cell lines include the SV40-transformed monkey kidney CV1 cell line (COS-7); human embryonic kidney cell lines (e.g., 293 or 293 cells as described in [Graham et al., J. Gen Virol. 36:59 (1977)]); baby hamster kidney cells (BHK); mouse Sertoli cells (e.g., TM4 cells as 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 (Hep G2); mouse mammary tumor cells (MMT 060562); e.g., [Mather et al., Annals NY Acad. Sci. 383:44-68] Examples include TRI cells, MRC 5 cells, and FS4 cells, as described in (1982). Other useful mammalian host cell lines include Chinese hamster ovary (CHO) cells - CHO cells (Urlaub et al., Proc. Natl. Acad. Sci. USA 77:4216 (1980)), including the aforementioned DHFR, and myeloma cell lines, such as Y0, NS0, and Sp2 / 0. For the study of specific mammalian host cell lines suitable for antibody production, see, for example, the literature [Yazaki and Wu, Methods in Molecular Biology, Vol. 248 (BKC Lo, ed., Humana Press, Totowa, NJ), pp. 255-268 (2003)].Examples of suitable mammalian host cell lines include the COS-7 cell line of monkey kidney cells (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 (ATCC CCL 70) derived from the African green monkey kidney cell line CV1, as described in the literature [McMahan et al., 1991, EMBO J. 10:2821].

[0046] In addition to prokaryotes, eukaryotic microorganisms such as filamentous fungi or yeasts, including fungal and yeast strains whose glycosylation pathways are humanized and induce the production of antibodies with partially or completely human-type glycosylation patterns, are also suitable cloning or expression hosts for antibody-encoding vectors. See references [Gerngross, Nat. Biotech. 22:1409-1414 (2004)] and [Li et al., Nat. Biotech. 24:210-215 (2006)]. Host cells suitable for the expression of glycosylated antibodies are derived from multicellular organisms (invertebrates and vertebrates). Examples of invertebrate cells include plant and insect cells. In particular, various baculovirus strains that can be used in combination with insect cells for the phenotypic infection of Spodoptera frugiperda cells have been identified. Cloning and expression vectors suitable for use with bacteria, fungi, yeast, and mammalian cell hosts have been described by Pouwels et al. [Cloning Vectors: A Laboratory Manual, (Elsevier, NY, 1985)].

[0047] Transformed cells can be cultured under conditions that promote polypeptide expression, and the polypeptide can be recovered by standard protein purification procedures. One of these purification procedures involves, for example, the use of affinity chromatography on a matrix having all or part of matrix-bound PD-L1 (e.g., the extracellular domain). The polypeptides considered for use in this application include substantially homogeneous recombinant mammalian anti-PD-L1 antibody polypeptides that are substantially free of endogenous contaminants. Therefore, antibodies can be produced 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 in this application is provided. Such nucleic acid may encode an amino acid sequence containing VL of the antibody and / or an amino acid sequence containing VH of the antibody (e.g., the light chain and / or heavy chain of the antibody). In additional embodiments, one or more vectors (e.g., expression vectors) containing these nucleic acids are provided. In additional embodiments, host cells containing these nucleic acids are provided. In one such embodiment, the host cells include (e.g., transformed into): (1) a vector containing nucleic acids encoding an amino acid sequence containing VL of the antibody and an amino acid sequence containing VH of the antibody, or (2) a first vector containing nucleic acids encoding an amino acid sequence containing VL of the antibody and a second vector containing nucleic acids encoding an amino acid sequence containing VH of the antibody. In one embodiment, the host cells are eukaryotes, for example, Chinese hamster ovary (CHO) cells or lymphoid cells (e.g., Y0, NS0, Sp20 cells). In one embodiment, a method for producing an anti-PD-L1 antibody is provided, comprising the steps of culturing host cells containing a nucleic acid encoding the antibody as provided above under appropriate conditions for antibody expression, and optionally recovering the antibody from the host cells (or host cell culture medium).

[0048] The proteins disclosed herein can also be produced using cell translation systems. For this purpose, the nucleic acids encoding polypeptides need to be modified to enable the generation of mRNA by in vitro transcription and to enable cell-free translation of mRNA in the specific cell-free system used (eukaryotes, e.g., mammalian or yeast cell-free translation systems, or prokaryotes, e.g., bacterial cell-free translation systems). Furthermore, PD-L1-binding polypeptides can also be produced by chemical synthesis (for example, by the method described in the literature [Solid Phase Peptide Synthesis, 2nd ed., 1984, The Pierce Chemical Co., Rockford, III.]). Additionally, protein modification can also be performed by chemical synthesis.

[0049] The polypeptides of the present invention can be purified by protein separation / purification methods commonly known in the field of protein chemistry. Non-limiting examples include extraction, recrystallization, salting out (e.g., salting out with ammonium sulfate or sodium sulfate), centrifugation, dialysis, ultrafiltration, adsorption chromatography, ion exchange chromatography, hydrophobic chromatography, normal phase chromatography, reverse phase chromatography, gel filtration, gel permeation chromatography, affinity chromatography, electroporation, back partitioning, or any combination thereof. After purification, the polypeptides can be exchanged for various buffers or concentrated by various known methods, including but not limited to filtration and dialysis. 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 exact purity level, the polypeptide is pure enough to be used as a pharmaceutical product. Antigen-binding proteins (e.g., antibodies, antibody fragments, antibody derivatives, antibody mutains, and antibody variants) are polypeptides that bind to PD-L1 (preferably human PD-L1). Antigen-binding proteins include antigen-binding proteins that inhibit the biological activity of PD-L1. Antigen-binding proteins can be produced by any of the many known techniques and screened for desired properties. Certain techniques include the steps of isolating a nucleic acid encoding a polypeptide chain (or a portion thereof) of an antigen-binding protein of interest (e.g., an anti-PD-L1 antibody), and manipulating the nucleic acid by recombinant DNA techniques. The nucleic acid can be fused with another nucleic acid of interest, or modified (e.g., by adding, deleting, or substituting one or more amino acid residues) using mutagenesis or other conventional techniques.

[0050] Single-chain antibodies can be formed by linking heavy-chain and light-chain variable domain (Fv region) fragments via an amino acid bridge (short peptide linker) to generate a single polypeptide chain. The single-chain Fv (scFv) was produced by fusing DNA encoding a peptide linker between DNA encoding two variable domain polypeptides (VL and VH). The resulting polypeptides can refold 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 polypeptides containing different VL and VH, anyone can form multimer scFv that bind to different epitopes (Kriangkum et al., 2001, Biomol. Eng. 18:31-40). Techniques developed for the production of single-chain antibodies include those described in U.S. Patent No. 4,946,778, and references [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].

[0051] A technique for inducing various subclasses or isotypes of antibodies from an antibody of interest, i.e., subclass switching, is known. Therefore, IgG antibodies can be derived from, for example, IgM antibodies, and conversely, IgM antibodies can be derived from IgG antibodies. Such techniques enable the production of novel antibodies that possess the antigen-binding properties of a specific antibody (parent antibody) but have biological properties associated with a different antibody isotype or subclass than the parent antibody. Recombinant DNA technology can be utilized. Cloned DNA encoding a specific antibody polypeptide, for example, DNA encoding the invariant domain of an antibody of the desired isotype, is used in these procedures (Lantto et al., 2002, Methods Mol. Biol. 178:303-16). Furthermore, if IgG4 is preferred, it may be preferable to introduce a point mutation (CPSCP → CPPCP) in the hinge region of the IgG4 antibody to reduce the tendency to form inter-H chain disulfide bonds that could cause heterogeneity (Bloom et al., 1997, Protein Science 6:407).

[0052] Pharmaceutical composition and administration Another aspect of the present invention provides pharmaceutical compositions comprising a PD-L1 inhibitor as a neoadjuvant therapy disclosed herein. Such pharmaceutical compositions can be formulated with pharmaceutically acceptable suitable carriers, excipients, buffers, and other substances that provide appropriate transport, delivery, tolerability, etc. Many suitable formulations are listed in prescription collections known to all pharmacists: Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA. These formulations include, for example, acids, pastes, ointments, jellies, waxes, oils, lipids, (LIPOFECTIN) TMExamples include lipid (cationic or anionic)-containing vesicles, DNA conjugates, anhydrous absorbent pastes, oil-in-water and water-in-oil emulsions, emulsion carbowaxes (polyethylene glycol of various molecular weights), semi-solid gels, and semi-solid mixtures containing carbowaxes. See also [Powell et al., "Compendium of excipients for parenteral formulations" PDA, J Pharm Sci Technol 52:238-311 (1998)].

[0053] The dose of a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody) varies depending on the age and weight of the subject, the target disease, the pathology, and the route of administration. When the PD-L1 inhibitor of the present invention is used to treat or inhibit the growth of gastric cancer, esophageal cancer, or liver cancer, the PD-L1 inhibitor can be administered as a single or multiple dose at a dose of approximately 0.1 mg to approximately 100 mg / kg body weight. The frequency and duration of treatment can be adjusted according to the severity of the disease. In some embodiments, the PD-L1 inhibitor of the present invention can be administered as an initial dose of at least approximately 0.1 mg to approximately 800 mg, approximately 1 mg to approximately 1000 mg, approximately 1 mg to approximately 800 mg, approximately 5 mg to approximately 500 mg, or approximately 10 mg to approximately 400 mg. In some embodiments, a second or more doses of the PD-L1 inhibitor may be administered after the initial dose, with subsequent doses being approximately the same as or less than the initial dose, and these subsequent doses are administered at intervals of 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 administered at intervals of at least 1 week, at least 2 weeks, at least 3 weeks, or at least 4 weeks.

[0054] Various delivery systems are known and can be used to administer the pharmaceutical composition of the present invention, such as liposome encapsulation, 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 delivery include, but are not limited to, intradermal, transdermal, intramuscular, intravenous, subcutaneous, intranasal, epidural, and oral routes. The composition can be administered by any convenient route, such as injection or bolus injection, by absorption through epithelial or mucocutaneous linings (e.g., oral mucosa, rectal and intestinal mucosa), and may be administered together with other biologically active substances. Pharmaceutical compositions can be delivered in the form of vesicles, particularly liposomes (see, for example, the reference [Langer (1990) Science 249:1527-1533]).

[0055] The use of nanoparticles for delivering the PD-L1 inhibitor of the present invention is also considered in this application. Antibody-conjugated nanoparticles can be used for both therapeutic and diagnostic applications. Antibody-conjugated nanoparticles and methods for their manufacture and use are described in detail in the literature [Arruebo, which is an engineered tracrRNA, al., 2009, "Antibody-conjugated nanoparticles for biomedical applications," J. Nanomat., Vol. 2009, Article ID 439389, 24 pages]. Nanoparticles can be developed and conjugated to antibodies contained in pharmaceutical compositions against target cells. Nanoparticles for drug delivery are described, for example, in U.S. Patent No. 8,257,740 or No. 8,246,995. In some situations, pharmaceutical compositions can be delivered in the form of 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 may be positioned near the target of the composition, so that only a portion of the systemic dose may be required. Injectable formulations may include dosage forms for intravenous, subcutaneous, intracranial, and intramuscular injection, as well as intravenous infusion. Such injectable formulations can be manufactured by known methods. In one embodiment, the injectable formulation may be a histidine solution.

[0056] The pharmaceutical composition of the present invention can be delivered subcutaneously or intravenously using a standard needle and syringe. For subcutaneous delivery, a pen delivery device can be appropriately used to deliver the pharmaceutical composition of the present invention. Such a pen delivery device may be reusable or disposable. Reusable pen delivery devices typically use a replaceable cartridge containing the pharmaceutical composition. Once all of 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 delivery device can then be reused. Disposable pen delivery devices do not have replaceable cartridges. Instead, disposable pen delivery devices have the pharmaceutical composition pre-filled in the device's storage compartment. Once the pharmaceutical composition from the storage compartment is depleted, the empty device is discarded. Advantageously, the above-mentioned oral or parenteral pharmaceutical compositions are prepared in unit dose forms appropriate to the dose of the active ingredient. Examples of such unit dose forms include tablets, pills, capsules, injections (ampoules), and suppositories. The antibody content is typically about 5 mg to 1000 mg per unit dose form, for example, about 5 mg to 600 mg, about 5 mg to 350 mg, or about 10 mg to 300 mg.

[0057] In certain embodiments, the present invention provides a pharmaceutical composition or formulation comprising a therapeutically effective amount of a PD-L1 inhibitor (e.g., IMC-001 or its bioequivalent) and a pharmaceutically acceptable carrier. Non-limiting examples of pharmaceutical compositions comprising the anti-PD-L1 antibody provided herein that are available in the context of the present invention are disclosed in U.S. Patent Publication No. 2019 / 0040137. Furthermore, in another aspect of the present invention, a kit is provided comprising a PD-L1 inhibitor (e.g., IMC-001 or its bioequivalent) for therapeutic use as described herein. The kit typically includes a label and instructions for use indicating the intended use of the kit's contents. As used herein, the term “label” includes any written or recorded material on, within, or accompanying to the kit, or otherwise attached to the kit. In one embodiment, a kit is provided for treating a patient with upper gastrointestinal cancer (e.g., gastric cancer, esophageal cancer, or liver cancer), comprising (a) a therapeutically effective dose of a PD-L1 inhibitor (e.g., IMC-001 or its bioequivalent), and (b) instructions for using the PD-L1 inhibitor in any manner disclosed herein.

[0058] Administration regimen and dosage In certain embodiments, the method of the present invention may include administering a therapeutically effective dose of a PD-L1 inhibitor (e.g., IMC-001 antibody or its bioequivalent) to an individual tumor one or more times, for example, one, two, three, four, five, six, seven, eight, nine, or ten times. For example, a therapeutic administration regimen may include administering one or more doses of the PD-L1 inhibitor at intervals of about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 12 weeks, about 1 month, about 2 months, about 3 months, or about 4 months, at longer intervals, or as needed, as long as a therapeutic response is achieved. In certain embodiments, the PD-L1 inhibitor is administered in one or more treatment cycles, e.g., one, two, three, four, five, six, seven, eight, nine, or ten treatment cycles. Methods in these aspects include administering one or more neoadjuvant therapy cycles to the required subjects and selectively administering one or more adjuvant therapy cycles, each treatment cycle including the administration of a PD-L1 inhibitor (e.g., IMC-001 or its bioequivalent) one, two, three, four, five, six, seven, eight, nine, ten, or more times.

[0059] In one embodiment, as neoadjuvant therapy, the PD-L1 inhibitor is administered once, twice, three times, four times, five times, or six times before surgical resection of the tumor. In one embodiment, as neoadjuvant therapy, the PD-L1 inhibitor is administered two to six times before surgical resection of the tumor. In one embodiment, as neoadjuvant therapy, the PD-L1 inhibitor is administered a total of two times (two treatment cycles) before surgical resection of the tumor. In one embodiment, as neoadjuvant therapy, the PD-L1 inhibitor is administered two to six times (six treatment cycles) before surgical resection of the tumor. In one embodiment, as neoadjuvant therapy, the PD-L1 inhibitor is administered once approximately every week, every two weeks, or every three weeks. In one embodiment, as neoadjuvant therapy, the PD-L1 inhibitor is administered at intervals of 10 to 18 days, preferably every 14 days. In certain embodiments, each dose of the PD-L1 inhibitor includes 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 per kg of the patient's body weight. In certain embodiments, each dose of the PD-L1 inhibitor is 20 mg / kg. In some embodiments, as neoadjuvant therapy, the PD-L1 inhibitor is administered at a dose of 20 mg / kg for a total of two doses at 10-18 day intervals prior to surgical resection of the tumor (2 cycles). The amount of PD-L1 inhibitor (e.g., IMC-001 or its bioequivalent) administered to a subject according to the method disclosed herein is typically a therapeutically effective dose. As used herein, the term “therapeutically effective dose” means an amount of PD-L1 inhibitor administered as neoadjuvant therapy before performing surgery planned to treat gastric cancer, esophageal cancer, or liver cancer, that achieves one or more of the following: compared to a subject treated by surgical resection alone or an untreated subject, respectively: (a) increased inhibition of tumor growth or tumor necrosis, tumor reduction and / or tumor disappearance; (b) reduced symptoms or signs of cancer, e.g., reduced severity or duration of tumor lesions; (c) delayed tumor growth and development; (d) inhibition of tumor metastasis; (e) prevention of recurrence of tumor growth; and / or (f) increased survival rate of a subject with cancer.

[0060] In certain embodiments, the therapeutically effective dose of the PD-L1 inhibitor (e.g., IMC-001 or its bioequivalent) may be approximately 0.05 mg to approximately 1000 mg, approximately 1 mg to approximately 800 mg, approximately 5 mg to approximately 600 mg, approximately 10 mg to approximately 550 mg, approximately 50 mg to approximately 400 mg, approximately 75 mg to approximately 350 mg, or approximately 100 mg to approximately 300 mg of antibody. For example, in various embodiments, the amount of PD-L1 inhibitor is approximately 0.05 mg, approximately 0.1 mg, approximately 1.0 mg, approximately 1.5 mg, approximately 2.0 mg, approximately 5 mg, approximately 10 mg, approximately 15 mg, approximately 20 mg, approximately 30 mg, approximately 40 mg, approximately 50 mg, approximately 60 mg, approximately 70 mg, approximately 80 mg, approximately 90 mg, approximately 100 mg, approximately 110 mg, approximately 120 mg, approximately 130 mg, approximately 140 mg, approximately 150 mg, approximately 160 mg, approximately 170 mg, approximately 180 mg, approximately 1 90mg, about 200mg, about 210mg, about 220mg, about 230mg, about 240mg, about 250mg, about 260mg, about 270mg, about 280mg, about 290mg, about 300mg, about 310mg, about 320mg, About 330mg, about 340mg, about 350mg, about 360mg, about 370mg, about 380mg, about 390mg, about 400mg, about 410mg, about 420mg, about 430mg, about 440mg, about 450mg, about 460mg , about 470mg, about 480mg, about 490mg, about 500mg, about 510mg, about 520mg, about 530mg, about 540mg, about 550mg, about 560mg, about 570mg, about 580mg, about 590mg, about 600 mg, about 610mg, about 620mg, about 630mg, about 640mg, about 650mg, about 660mg, about 670mg, about 680mg, about 690mg, about 700mg, about 710mg, about 720mg, about 730mg, about 74 The amounts are 0 mg, approximately 750 mg, approximately 760 mg, approximately 770 mg, approximately 780 mg, approximately 790 mg, approximately 800 mg, approximately 810 mg, approximately 820 mg, approximately 830 mg, approximately 840 mg, approximately 850 mg, approximately 860 mg, approximately 870 mg, approximately 880 mg, approximately 890 mg, approximately 900 mg, approximately 910 mg, approximately 920 mg, approximately 930 mg, approximately 940 mg, approximately 950 mg, approximately 960 mg, approximately 970 mg, approximately 980 mg, approximately 990 mg, or approximately 1000 mg.

[0061] The amount of PD-L1 inhibitor (e.g., IMC-001 or its bioequivalent) contained in each dose can be expressed in mg of antibody per kg of body weight of the subject (i.e., mg / kg). In certain embodiments, the PD-L1 inhibitor used in the methods disclosed herein can be administered to the subject in doses of about 0.0001 mg to about 100 mg / kg relative to the subject's body weight. In certain embodiments, the anti-PD-L1 antibody can be administered to the subject in doses of about 0.1 mg / kg to about 30 mg / kg relative to the subject's body weight. In certain embodiments, the methods of the present invention include administering a PD-L1 inhibitor (e.g., anti-PD-L1 antibody IMC-001 or its bioequivalent) in 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 relative to the subject's body weight. In certain embodiments, the individual dose content of the 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 is 20 mg / kg, a subtherapeutic dose may be less than 20 mg / kg, e.g., 18 mg / kg, 16 mg / kg, 14 mg / kg, 12 mg / kg, or 10 mg / kg. As defined herein, a “subtherapeutic dose” means an amount of PD-L1 inhibitor that does not produce a therapeutic effect on its own. However, in certain embodiments, the PD-L1 inhibitor is administered multiple times in subtherapeutic doses to achieve a therapeutic effect collectively in the subject.

[0062] In certain embodiments, each dose contains a PD-L1 inhibitor (e.g., IMC-001 or its bioequivalent) at a dose of 0.1 to 30 mg / kg (e.g., 5 mg / kg, 10 mg / kg, 15 mg / kg, 20 mg / kg, or 25 mg / kg) based on the patient's body weight. In some other embodiments, each dose contains 5 to 600 mg of the PD-L1 inhibitor, for example, 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 the PD-L1 inhibitor. In one embodiment, the therapeutically effective dose of a PD-L1 inhibitor (e.g., IMC-001 or its bioequivalent) is 20 mg / kg administered intravenously as neoadjuvant therapy before planned surgery for gastric, esophageal, or liver 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 as postoperative adjuvant therapy.

[0063] In certain embodiments, surgical resection of the tumor is performed 11 days or within 84 days of the last dose of the PD-L1 inhibitor. In certain embodiments, surgical resection of the tumor is performed 11 days or within 84 days of the last dose of the PD-L1 inhibitor. In certain embodiments, surgical resection of the tumor is performed between 11 and 84 days of the last dose of the PD-L1 inhibitor. In certain embodiments, surgical resection of the tumor is performed between 11 and 42 days of the last dose of the PD-L1 inhibitor. In certain embodiments, surgical resection of the tumor is performed between 18 and 35 days, or between 21 and 35 days, of the last dose of the PD-L1 inhibitor. In a particular embodiment, the anti-PD-L1 antibody is administered twice at a dose of 20 mg / kg at intervals of 10 to 18 days or 2 weeks prior to surgical resection of the tumor in the patient, with the surgical resection of the tumor taking place between 11 and 42 days after the last dose of the anti-PD-L1 antibody.

[0064] All patents and references cited herein are incorporated herein by reference in their entirety. The present invention will be described in more detail below with reference to the following examples. However, the following examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples. [Examples]

[0065] Example 1. Clinical trial of IMC-001 as neoadjuvant therapy for the treatment of resectable gastric, esophageal, and liver cancers. 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 IgG1 containing a heavy chain with the amino acid sequence described in SEQ ID NO: 3 and a light chain with the amino acid sequence described in SEQ ID NO: 4. See Table 1 below for the amino acid sequence information of IMC-001.

[0066] [Table 1]

[0067] [Table 2]

[0068] IMC-001 was produced in CHO cells as H6B1L-EM, a variant of the wild-type parental antibody H6B1L. For further details, please refer to International Publication No. WO 2017 / 132562 A1. This 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.

[0069] Research objectives One objective of this study is to evaluate the clinical activity of IMC-001 as neoadjuvant therapy in patients with resectable gastric, esophageal, and hepatocellular carcinoma lesions. basis Neoadjuvant immunotherapy offers potential biological advantages, including less immunosuppressive TME, lower disease burden, and antitumor immune priming in the presence of primary tumors. Gastrointestinal cancers, including gastroesophageal cancer and hepatocellular carcinoma, are among the cancers in which immune checkpoint inhibitors (ICIs) offer survival benefits in palliative care. IMC-001 is a fully human anti-PD-L1 recombinant monoclonal antibody that stimulates ADCC by maintaining the Fc effector and demonstrated a favorable safety profile at up to 20 mg / kg IV every two weeks in a Phase I trial (n=15), showing promising preliminary efficacy in patients with solid tumors who had received multiple treatments (1 partial remission, 4 stable disease) (Keam B, et al, Invest New Drugs 2021;39:1624-32).

[0070] Study Endpoints The primary endpoint of this study is to evaluate the major pathologic response rate in surgical tissue after administration of the preoperative pre-emptive immune checkpoint inhibitor IMC-001. Secondary endpoints include evaluation of the feasibility and safety of planned surgical delay with the preoperative pre-emptive therapy IMC-001, evaluation of the R0 resection rate, evaluation of the clinical tumor response rate according to RECIST v1.1, evaluation of the clinical disease control rate according to RECIST v1.1, evaluation of progression-free survival, evaluation of relapse-free survival, evaluation of overall survival, and evaluation of cancer progression / recurrence rate and the characteristics of cancer progression / recurrence. On the other hand, the exploratory objective of this study is to obtain subject samples (tissue, blood, feces) before and after administration of the immune checkpoint inhibitor IMC-001, construct a biomarker cohort, perform immunoprofiling and genomic analysis, and discover biomarkers that predict therapeutic efficacy.

[0071] Experimental Design Clinical trial participants who met the selection / exclusion criteria were assigned to one of three cohorts: gastric cancer, esophageal cancer, or hepatocellular carcinoma. In each cancer type cohort, two cycles of IMC-001 (consisting of 1 cycle = 2 weeks) were administered as pre-treatment, followed by curative surgery, and then follow-up observation. After curative surgery, adjuvant therapy was administered as needed, according to standard clinical practice guidelines and the stage of the disease. The study included patients with no prior treatment history for resectable localized gastric cancer, esophageal cancer, or hepatocellular carcinoma, and a total of 48 patients (16 per cancer cohort) were enrolled. Neoadjuvant therapy Unless there was unacceptable toxicity, clinical cancer progression, or withdrawal of consent, IMC-001 was administered twice at 2-week intervals as pre-treatment before surgical resection.

[0072] Collection of biomarker samples Tumor tissue, blood, and stool samples were collected before IMC-001 administration, blood and stool samples were collected after the first and second cycles of administration, and surgical tumor tissue, blood, and stool samples were collected after surgery. Tracking observation Based on the date of surgery, blood, stool, and tumor tissue samples were collected and monitored every three months for the first two years, every six months for the following three years, and only when cancer recurred. Participants in the clinical trial This study enrolled a total of 48 patients (16 from each cancer type cohort) with gastric adenocarcinoma, esophageal squamous cell carcinoma, or hepatocellular carcinoma that were suitable for curative surgical resection.

[0073] Selection criteria Disease selection criteria 1) Histologically confirmed localized gastric adenocarcinoma, esophageal squamous cell carcinoma, hepatocellular carcinoma, or hepatocellular carcinoma clinically diagnosed according to the AASLD (American Association for the Study of Liver Disease) guidelines. However, in the case of hepatocellular carcinoma that is clinically diagnoseable according to the AASLD guidelines, histological examination will not be performed. 2) Gastric adenocarcinoma, esophageal squamous cell carcinoma, or hepatocellular carcinoma that is expected to be curatively resectable. - Gastric adenocarcinoma: Clinical stage ≥ T2 or positive lymph node metastasis (AJCC 8) th ) - Esophageal squamous cell carcinoma: Clinical stage ≥ T1b or positive lymph node metastasis (AJCC 8) th ) - Hepatocellular carcinoma: A single hepatocellular carcinoma confined to the liver, or three or fewer hepatocellular carcinomas confined to the liver. Invasion of the hepatic portal vein, hepatic vein, and biliary tract may occur, but there must be no invasion of the main portal trunk. 3) Hematological, hemochemical, and major organ function requirements must meet the following criteria (confirmed within 7 days prior to the first dose of the study drug). - Absolute neutrophil count ≧1,000 / μL - Platelet count ≥75,000 / μL - Serum total bilirubin ≤ 1.5 x upper limit of normal (ULN) (For subjects with Gilbert's syndrome, total bilirubin ≤ 3.0 x ULN) - AST (aspartate aminotransferase) or ALT (alanine aminotransferase) ≤ 2.5 x ULN; alkaline phosphatase ≤ 2.5 x ULN - Serum creatinine ≤ 1.5 x upper limit of normal or creatinine clearance ≥ 50 mL / minute (However, creatinine clearance is first calculated using the Cockcroft-Gault formula based on sex. If the value is less than 50 mL / min, a 24-hour urine sample is collected and tested. If the creatinine clearance is 50 mL / min or higher, registration is possible.)

[0074]

number

[0075]

number

[0076] - Urine protein-creatinine ratio (UPC) ≤ 1 (however, if UPC > 1, registration is possible if the 24-hour urine protein sample is less than 2g). - In addition, in the case of hepatocellular carcinoma, sufficient liver function must be confirmed to be Child-Pugh grade A (see Table 4) and encephalopathy grade 0 (see Table 3) (see Table 2).

[0077] [Table 3]

[0078] [Table 4]

[0079] [Table 5]

[0080] 4) A measurable or evaluable lesion based on RECIST (Response Evaluation Criteria in Solid Tumors) version 1.1 (see reference [Eisenhauer EA, Therasse P, Bogaerts J, Schwartz LH, Sargent D, Ford R, et al. New response evaluation criteria in solid tumours: revised RECIST guideline (Version 1.1). Eur J Cancer. 2009; 45:228 247]). 5) Ability to provide tumor tissue samples deemed appropriate for biomarker analysis (in the case of hepatocellular carcinoma, registration is possible even without tumor tissue samples taken before administration of the investigational drug). General selection criteria 6) Must be 19 years of age or older. 7) ECOG (Eastern Cooperative Oncology Group) Performance Status: 0-1 (See Table 5)

[0081] [Table 6]

[0082] 8) You have signed the consent form. 9) In the case of men and women of childbearing age, they must be able to adhere to appropriate contraception during treatment and for three months after the last dose of treatment.

[0083] Exclusion criteria Participants who met any of the following criteria were excluded from this study. Tumor-related exclusion criteria 1) Unresectable or metastatic disease. 2) If you have previously received treatment for gastric adenocarcinoma, esophageal squamous cell carcinoma, or hepatocellular carcinoma. However, in the case of hepatocellular carcinoma, even if you have previously received treatment for a local lesion, you may be enrolled in the trial if more than 6 months have passed since treatment, the treated area has progressed, or a new lesion has developed outside the previously treated area, is resectable, and meets other selection / exclusion criteria. 3) Patients with a history of another cancer within three years prior to the start of the investigational treatment. However, patients with conditions that have little impact on the patient's prognosis, such as carcinoma in situ or papillary thyroid carcinoma, may be enrolled at the discretion of the principal investigator. 4) History of hepatic encephalopathy 5) Clinically significant ascites as defined below: - If ascites is found during the physical examination at the time of screening or - Past ascites requiring treatment and ongoing prevention, or current ascites requiring treatment.

[0084] Exclusion Criteria for Investigational Drugs 6) A history of active autoimmune disease requiring systemic treatment (i.e., disease modifiers, corticosteroids, or immunosuppressants) within the past two years. Alternative therapies (e.g., thyroxine, insulin, or physiological corticosteroid replacement therapy due to adrenal or pituitary dysfunction) are not considered systemic treatment and are acceptable. 7) Subjects diagnosed with immunodeficiency, or subjects receiving chronic systemic steroid therapy (a dose exceeding the equivalent of 10 mg of prednisone per day) or any other form of immunosuppressive therapy within 7 days prior to the first dose of the study drug. 8) You have a history of (non-infectious) interstitial pneumonia requiring steroid treatment, or you currently have interstitial pneumonia. 9) A history of 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 costimulation or gateway pathways. 10) Known severe hypersensitivity or anaphylaxis to recombinant proteins, including monoclonal antibodies.

[0085] General exclusion criteria 11) Presence of active infection requiring systemic treatment 12) The Principal Investigator has, or has currently had, a medical condition, treatment, or a history of abnormal clinical laboratory values ​​that could interfere with the results of the trial, prevent the participant from participating in the trial, or would not be in the participant's best interest to participate in the trial. 13) Pregnant women who have tested positive for pregnancy in urine or blood within 7 days prior to the first dose of the investigational drug. 14) You are pregnant or breastfeeding, or you plan to become pregnant or give birth during the scheduled clinical trial period, up to 90 days after the last dose of the investigational drug. 15) Having symptomatic congestive heart failure (i.e., class II or higher according to the New York Heart Association classification), or a history or current finding of clinically significant cardiac arrhythmias requiring treatment with antiarrhythmic drugs other than beta-blockers or digoxin, and / or having a history of conduction disorder (excluding atrial fibrillation and paroxysmal ventricular tachycardia), active coronary artery disease, unstable angina, new-onset angina within 3 months prior to enrollment, or myocardial infarction within 6 months prior to enrollment. 16) The patient has a known history of exposure to human immunodeficiency virus (HIV) (HIV 1 / 2 antibodies). 17) Subjects with known active hepatitis B (detection of hepatitis B surface antigen HBsAg or HBV DNA) or hepatitis C (detection of HCV RNA). However, for subjects with hepatitis B, enrollment is permitted if HBV DNA is less than 500 IU / mL (or 2500 copies / mL) at the time of screening. Subjects in whom HBsAg or HBV DNA is detected must be managed according to treatment guidelines. Subjects receiving antiviral drugs at the time of screening must have received treatment for at least two weeks prior to enrollment and must continue treatment for six months after treatment with the study drug. Patients positive for HCV antibodies can only be enrolled if their PCR test is negative for HCV RNA (exception: patients with hepatocellular carcinoma can be enrolled regardless of whether they are HCV RNA positive or not). 18) A history of allogeneic tissue or solid organ transplantation. 19) The patient received a live vaccine within 28 days prior to the first dose of the investigational drug.

[0086] Experimental treatment (pre-operative IMC-001 treatment) Preparation and administration methods of investigational drugs IMC-001 was provided as an injectable preparation, filled 10 mL of sterile glass vials with histidine solution at concentrations of 10 mg / mL and 60 mg / mL. IMC-001 was diluted with 250 mL of 0.9% sodium chloride and administered by intravenous infusion over 60 minutes (±10 minutes). During administration of IMC-001, subjects were carefully monitored for infusion reactions. Administration of prior therapies was permitted, and at the discretion of the principal investigator, prior therapies such as antihistamines, antipyretics, and / or analgesics (e.g., acetaminophen) were administered. In the event of an infusion-related reaction, subjects were managed according to treatment guidelines. Vital signs (blood pressure, pulse rate, respiratory rate, body temperature) were measured within 60 minutes prior to infusion. Vital signs were measured every 15 minutes (±10 minutes) during infusion or 30 minutes (±10 minutes) after infusion, if clinically necessary. If a subject experienced an infusion-related reaction during the initial infusion, vital signs were measured during the infusion and 30 minutes (±10 minutes) after the infusion.

[0087] Initial dose and administration schedule The dose of IMC-001 in this study was 20 mg / kg, which was determined as the recommended dose (RP2D) for Phase II in the ongoing Phase I study. It was administered intravenously on day 1 of a 14-day cycle (±4 days). Dose calculations were based on body weight assessed at baseline. If a subject's body weight was within 10% of the body weight used in previous dose calculations, the dose was not recalculated. All doses were rounded to the nearest milligram (mg) according to the standards of the clinical trial site. Total duration of administration Unless the patient dropped out early or withdrew consent due to unacceptable toxicity, cancer progression, or because it was determined that continuing treatment would be detrimental, a total of two cycles of preoperative IMC-001 were administered.

[0088] Surgery: Surgery was performed within four weeks of completion of the second cycle of the preceding therapy IMC-001 (from day 1 to day 42 of IMC-001 administration in the second cycle). However, the date of the visit to complete the preceding therapy preceded the date of surgery. At this time, the subjects had a white blood cell count exceeding 3,000 / μL, a platelet count exceeding 75,000 / μL, no grade 2 or higher toxicity from the investigational drug treatment, and were clinically capable of undergoing major surgery. Surgical procedures were performed in accordance with institutional treatment guidelines for gastric cancer, esophageal cancer, and hepatocellular carcinoma. Open surgery was the standard approach. Laparoscopic and robot-assisted surgery were permitted after consultation with the principal investigator, and these surgeries were only permitted if performed by a surgical team with sufficient expertise to be considered equivalent to the open surgery approach in terms of morbidity, mortality, and oncological outcomes. Laparoscopic surgery in the clinical trial could be considered after the surgical team reviewed the extent and resectableness of the tumor and the presence of distant metastases during the initial evaluation. Excision The extent of surgical resection was determined based on the location, size, and stage of the tumor. The surgical resection feasibility of the cancer was determined by the surgeon's intraoperative judgment. The following may be reasons why resection is not possible: - Distant metastasis - Tumors that invade major blood vessels and / or other organs - In cases where other radical resections are deemed difficult. Generally, if complete resection (R0) is not possible, the surgical team decides whether or not to perform non-curative surgery. Lymph node dissection The extent of lymph node dissection was determined based on institutional treatment guidelines for gastric cancer, esophageal cancer, and hepatocellular carcinoma.

[0089] Postoperative treatment Adjuvant chemotherapy after curative resection, as well as treatment for patients who underwent non-curative resection or were unable to undergo resection, were carried out in accordance with the institution's guidelines. Drugs and treatments that are prohibited from being used together. The following drugs and treatments were prohibited during the study (except when used to treat adverse events related to the study drug): - During the administration of prior anticancer drugs in this study, anticancer therapies for cancer treatment other than the investigational drug (including anticancer chemotherapy, hormone therapy, immunotherapy, radiation therapy, biological anticancer therapy, or herbal medicine) - Other investigational drug treatments within 28 days prior to the start of administration of the investigational drug used in this clinical trial and during administration of the investigational drug. - Attenuated live vaccine (e.g., FluMist) within 28 days prior to the start of investigational drug administration, during investigational drug administration, and for 5 months after the last investigational drug dose. - Systemic immunosuppressants (cyclophosphamide, azathioprine, methotrexate, thalidomide, etc.) - Systemic corticosteroids: Generally, the use of corticosteroids exceeding 10 mg / day of prednisone or similar doses is not permitted. Corticosteroids were temporarily allowed to exceed 10 mg / day when medically necessary, such as to suppress symptoms of adverse events like infusion reactions or irAEs, or when administered before computed tomography (CT) in subjects with a history of allergy to intravenous contrast agents. If the corticosteroid dose could not be reduced to a prednisone dose of 10 mg / day or less within 12 weeks of starting steroid administration, the principal investigator promptly began discussions regarding the continuation of the subject's investigational drug treatment.

[0090] Drugs and treatments that can be used in combination The following medications and treatments were approved: - Oral contraceptives - Hormone replacement therapy - Inactivated influenza vaccine - Megestrol acetate administered as an appetite stimulant. - Topical, intraocular, intra-articular, intranasal, and inhaled corticosteroids - Corticosteroid replacement therapy with less than 10 mg / day of prednisone - Corticosteroids used temporarily for prevention (for example, contrast agent allergy) The use of other medications necessary for subject management was at the discretion of the principal investigator. If nausea, vomiting, or diarrhea occurred, effective symptomatic treatment was initiated. Antiemetics were administered to subjects in accordance with the site guidelines or other guidelines (e.g., ASCO guidelines). For the treatment of febrile neutropenia, hematopoietic growth factors, i.e., G-CSF or GM-CSF, were used for primary prevention purposes, in accordance with the site guidelines or other guidelines (e.g., ASCO guidelines). For the treatment of infusion reactions related to the study drug, paracetamol (acetaminophen), pethidine (meperidine), chlorpheniramine, or other antihistamines were used in accordance with the site guidelines.

[0091] Severity assessment of adverse events The severity of adverse events was assessed using the adverse event severity scale based on NCI CTCAE (Common Terminology Criteria) v4.03 (http: / / ctep.cancer.gov / protocolDevelopment / electronic_applications / ctc.htm#ctc_40). For adverse events not included in the NCI CTCAE, Table 6 was used to assess their severity. Heart failure was classified according to the NYHA classification criteria.

[0092] [Table 7]

[0093] Example 2. Clinical trial results of IMC-001 as neoadjuvant therapy in the treatment of resectable GC, ESCC, and HCC. safety With the exception of elevated G3AST / ALT (n=3, 6%), all treatment-related adverse events (TRAEs) were G1 or G2. TRAEs occurring in less than 5% of patients included hyperthyroidism (n=9, 18%), hypothyroidism (n=7, 14%), fatigue (n=7, 14%), itching (n=6, 12%), skin rash (n=4, 8%), infusion-related reactions (n=3, 6%), and diarrhea (n=3, 6%). Clinical oncological response No patients experienced disease progression during neoadjuvant therapy. Of the 17 patients with measurable lesions according to RECIST 1.1, 3 (17.6%) achieved partial remission (PR) and 14 (82.4%) achieved stable disease (SD) (Figure 2). Of the 30 patients whose metabolic response could be evaluated according to the EORTC criteria, 7 (23.3%) showed metabolic partial remission (mPR), 20 (66.7%) showed metabolic stable disease (mSD), and 3 (10.0%) showed metabolic progressive disease (mPD) (Figure 3). Of the 3 patients with metabolic progressive disease, 2 showed an increase in tumor-infiltrating lymphocytes and tumor regression on surgical pathology.

[0094] surgery results All patients underwent radical resection (R0 resection) for stage I (n=36, 75.0%), stage II (n=6, 12.5%), and stage III (n=6, 12.5%). There were no reported surgical mortality rates or treatment-related surgical morbidities. Pathological reaction Although there were no major pathological reactions, 51% of patients showed varying degrees of tumor necrosis or fibrosis, and seven patients (15%) had less than 50% of tumor cells remaining (Figure 4). recurrence The median follow-up period was 31.4 months (range: 1.7 months to 41.2 months), with only 4 patients (8.3%) experiencing relapse and failing to reach the median progression-free survival (PFS) or relapse-free survival (RFS). conclusion IMC-001 neoadjuvant therapy is microsatellite-stable and demonstrated good tolerability and promising antitumor activity in patients with resectable GC, EC, and HCC.

Claims

1. A pharmaceutical composition for treating tumors or inhibiting tumor growth in patients with upper gastrointestinal cancer, The anti-PD-L1 antibody, in a therapeutically effective dose, specifically binds to PD-L1 and contains HCDR1, HCDR2, and HCDR3 from the heavy chain variable region (HCVR) of SEQ ID NO: 1, and LCDR1, LCDR2, and LCDR3 from the light chain variable region (LCVR) of SEQ ID NO:

2. A pharmaceutical composition comprising the antibody administered to a patient as neoadjuvant therapy prior to surgical resection of a tumor in the patient.

2. The pharmaceutical composition according to claim 1, wherein the upper gastrointestinal cancer is gastric 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 including HCDR1 of SEQ ID NO: 5, HCDR2 of SEQ ID NO: 6, and HCDR3 of SEQ ID NO: 7, and a light chain variable region including 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 includes the amino acid sequence of SEQ ID NO: 2, or includes an amino acid sequence that is 95% or more identical to SEQ ID NO:

2.

5. The pharmaceutical composition according to claim 4, wherein the amino acids at positions 3 and 5 of the light chain variable region are the same as the amino acids 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 an amino acid sequence that is 95% or more identical to that of 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 upper gastrointestinal cancer is resectable.

9. The pharmaceutical composition according to claim 1, wherein the 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 the upper gastrointestinal cancer is curative by surgical intervention.

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

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

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

15. The pharmaceutical composition according to claim 1, wherein, as the preoperative adjuvant therapy, an anti-PD-L1 antibody is administered once or more times before surgical resection of the tumor.

16. The pharmaceutical composition according to claim 15, wherein each of the aforementioned doses is administered at intervals of 10 to 18 days.

17. The pharmaceutical composition according to claim 16, wherein, as the preoperative adjuvant therapy, an anti-PD-L1 antibody is administered 2 to 6 times at intervals of 10 to 18 days prior to the surgical resection of the tumor.

18. The pharmaceutical composition according to claim 1, wherein an anti-PD-L1 antibody is administered in a dose of 10 mg / kg to 30 mg / kg as the aforementioned preoperative adjuvant therapy.

19. The pharmaceutical composition according to claim 18, wherein an anti-PD-L1 antibody is administered at a dose of 20 mg / kg as the aforementioned preoperative adjuvant therapy.

20. The pharmaceutical composition according to claim 1, wherein the surgical resection of the tumor is 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 its growth, (a) A step of screening patients for upper gastrointestinal cancer, (b) The step of administering a PD-L1 inhibitor to the patient in a therapeutically effective dose, (c) The step of surgically resecting the upper gastrointestinal cancerous tumor after step (b), A method wherein the PD-L1 inhibitor is an antibody that specifically binds to PD-L1 and 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 containing a PD-L1 inhibitor, along with instructions for use as neoadjuvant therapy to treat or inhibit tumor growth in patients with upper gastrointestinal cancer, The kit includes instructions for administering the PD-L1 inhibitor intravenously at a dose of 20 mg / kg two to six times at intervals of 10 to 18 days prior to surgical resection of the tumor.

Images