Neoadjuvant therapy using PD-l1 inhibitor for treating cancer

Abstract

The present invention relates to a method for treating tumors or inhibiting tumor proliferation, comprising administering, as a neoadjuvant therapy, a therapeutically effective amount of a programmed death-ligand 1 (PD-L1) inhibitor (for example, an anti-PD-L1 antibody) to a cancer patient in need thereof, followed by surgical resection. In particular, the present invention relates to a method for administering a PD-L1 inhibitor as a neoadjuvant therapy prior to surgery to patients with surgically resectable upper gastrointestinal cancer, for example, localized gastric cancer, esophageal cancer or liver cancer. It has been identified that the administration of a PD-L1 inhibitor, particularly an anti-PD-L1 antibody, exhibits an excellent therapeutic effect as a neoadjuvant therapy for patients with surgically resectable localized gastric cancer, esophageal cancer, and liver cancer.

Description

Neoadjuvant therapy using PD-L1 inhibitors for cancer treatment

[0001] The present invention relates to a method for treating a tumor or inhibiting tumor growth, comprising administering a therapeutically effective amount of a programmed death-ligand 1 (PD-L1) inhibitor (e.g., anti-PD-L1 antibody IMC-001 or a biological equivalent thereof) to a cancer patient as neoadjuvant therapy, followed by surgical resection. In particular, the present invention relates to a method for administering a PD-L1 inhibitor as neoadjuvant therapy before surgery to a patient with upper gastrointestinal cancer, e.g., localized gastric cancer, esophageal cancer, or liver cancer, which is amenable to surgical resection.

[0002] Sequence list

[0003] This application includes a sequence listing, which has been submitted electronically in XML format and is incorporated herein by reference in its entirety. A copy of said sequence listing, dated April 13, 2024, is named KC24060.xml and is 12.3 kilobytes in size.

[0004] Immune evasion, a key feature of cancer, has recently gained attention, and its mechanisms are being elucidated through advancements in tumor immunology. The immune checkpoint, a concept encompassing immunosuppressive regulatory molecules that prevent immune cells from attacking self-cells, is one of several mechanisms by which tumor cells evade immune surveillance, creating an immunosuppressive environment within the tumor.

[0005] Immunotherapy strategies that achieve anticancer effects by activating the suppressed antitumor immune system have attracted 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 types of cancer, establishing them as one of the standard immunotherapy cancer treatments and suggesting a new paradigm in cancer treatment (Pardoll DM: The blockade of immune checkpoints in cancer immunotherapy. Nat Rev Cancer 12:252-64, 2012).

[0006] There are research results in lung cancer that show that the therapeutic effect of immune checkpoint inhibitors increases as the number of mutations in the tumor increases, and in particular, the therapeutic effect increases as the number of clonal neoantigens increases, and that even if the number of total mutations is high, the therapeutic effect of PD-1 inhibitors is low when the number of subclonal neoantigens is high (McGranahan N, Furness AJ, Rosenthal R, et al: Clonal neoantigens elicit T cell immunoreactivity and sensitivity to immune checkpoint blockade. Science 351:1463-9, 2016).

[0007] Furthermore, considering that as cancer progresses, the number of subclonal neoantigens increases with tumor evolution and the immunosuppressive microenvironment develops, immune checkpoint inhibitor therapy may be 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).

[0008] Immune checkpoint inhibitors such as pembrolizumab, nivolumab, and ipilimumab have already been proven to be effective in treating unresectable locally advanced or metastatic gastric cancer, esophageal cancer, and hepatocellular carcinoma by improving survival rates or reducing tumor size.

[0009] Meanwhile, surgical resection is the treatment option for operable gastric, esophageal, and hepatocellular carcinomas. However, postoperative tumor recurrence is common, and early recurrences are also observed. Therefore, there is a critical need for safe and effective therapies for treating cancers that can be surgically treated.

[0010] The purpose of the present invention is to solve all of the problems of the above-mentioned prior art.

[0011] The present invention aims to provide a preoperative neoadjuvant therapy for patients with upper gastrointestinal cancer amenable to surgical resection, a cancer treatment method, a pharmaceutical composition, a kit, and a dosing regimen for cancer treatment.

[0012] The purpose of the present invention is not limited to the purposes mentioned above. The purpose of the present invention will become clearer from the following description and may be realized by the means and combinations thereof set forth in the claims.

[0013] A representative configuration of the present invention to achieve the above purpose is as follows.

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

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

[0016] In one embodiment, the upper gastrointestinal cancer may be gastric cancer, esophageal cancer, or liver cancer.

[0017] In one embodiment, the anti-PD-L1 antibody can comprise 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.

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

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

[0020] In one embodiment, the anti-PD-L1 antibody may comprise 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.

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

[0022] In one embodiment, the anti-PD-L1 antibody may retain Fc effector function capable of stimulating antibody dependent cellular cytotoxicity (ADCC).

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

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

[0025] In one embodiment, the anti-PD-L1 antibody as neoadjuvant therapy may be administered intravenously or subcutaneously.

[0026] In one embodiment, the upper gastrointestinal cancer may be resectable.

[0027] In one embodiment, the upper gastrointestinal cancer may be recurrent.

[0028] In one embodiment, the upper gastrointestinal cancer may be metastatic.

[0029] In one embodiment, the surgical goal for upper gastrointestinal cancer may be curative.

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

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

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

[0033] In one embodiment, the anti-PD-L1 antibody as neoadjuvant therapy may be administered one or more times, with each dose administered one week apart, two weeks apart, three weeks apart, four weeks apart, five weeks apart, or six weeks apart.

[0034] In one embodiment, the anti-PD-L1 antibody as neoadjuvant therapy may be administered more than once, with each dose administered two weeks apart.

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

[0036] In one embodiment, the anti-PD-L1 antibody as neoadjuvant therapy may be administered at a dose of 10 mg / kg to 30 mg / kg of body weight of the patient.

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

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

[0039] Another aspect of the present invention provides a kit comprising a PD-L1 inhibitor together with instructions for use of the PD-L1 inhibitor (e.g., an anti-PD-L1 antibody that specifically binds to PD-L1 and comprises HCDR1, HCDR2 and HCDR3 contained in the heavy chain variable region (HCVR) of SEQ ID NO: 1 and LCDR1, LCDR2 and LCDR3 contained in the light chain variable region (LCVR) of SEQ ID NO: 2) as neoadjuvant therapy for treating a tumor or inhibiting tumor growth in a patient with upper gastrointestinal cancer.

[0040] In one embodiment, the instructions for use included in the kit comprise administering a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody that specifically binds to PD-L1 and comprises HCDR1, HCDR2 and HCDR3 contained in the heavy chain variable region (HCVR) of SEQ ID NO: 1 and LCDR1, LCDR2 and LCDR3 contained in the light chain variable region (LCVR) of SEQ ID NO: 2) as neoadjuvant therapy to a patient with upper gastrointestinal cancer at a dose of 20 mg / kg administered once, twice, three times, four times, five times or six times at intervals of 10 to 18 days prior to surgical resection of the tumor.

[0041] 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 comprises HCDR1, HCDR2 and HCDR3 contained in a heavy chain variable region (HCVR) of SEQ ID NO: 1 and LCDR1, LCDR2 and LCDR3 contained in a 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 the steps of: (a) selecting a patient with upper gastrointestinal cancer; (b) administering a therapeutically effective amount of a PD-L1 inhibitor to the patient; and (c) surgically resecting the upper gastrointestinal cancer tumor after step (b).

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

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

[0044] Another aspect of the present invention provides a method for treating a tumor or inhibiting tumor growth, comprising the steps of: (a) selecting a patient with resectable liver 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 comprises HCDR1, HCDR2, and HCDR3 contained in a heavy chain variable region (HCVR) of SEQ ID NO: 1 and LCDR1, LCDR2, and LCDR3 contained in a light chain variable region (LCVR) of SEQ ID NO: 2); and (c) surgically resecting the liver cancer tumor after step (b).

[0045] Neoadjuvant therapy with anti-PD-L1 antibodies was well tolerated and showed promising antitumor activity in patients with microsatellite stable, resectable gastric, esophageal, and hepatocellular carcinomas. In one embodiment of the present invention, neoadjuvant therapy with IMC-001 before surgery resulted in low-grade treatment-emergent adverse events (TRAEs), partial remission (PR) or stable disease (SD) in patients with measurable lesions, and metabolic partial remission (mPR), metabolic stable disease (mSD), or metabolic progressive disease (mPD) in patients for whom metabolic response was evaluable. In addition, neoadjuvant therapy with IMC-001 before surgery resulted in variable degrees of tumor necrosis or fibrosis, and in some patients, residual viable tumor cells were reduced to less than 50%.

[0046] Figure 1 schematically illustrates the procedure of a clinical trial according to one embodiment of the present invention.

[0047] Figure 2 illustrates clinical tumor responses as a result of a clinical trial according to one embodiment of the present invention. PR indicates partial response, and SD indicates stable disease.

[0048] Figure 3 illustrates clinical tumor response as a result of a clinical trial according to one embodiment of the present invention. mPR indicates metabolic partial response, mSD indicates metabolic stable disease, and mPD indicates metabolic progressive disease.

[0049] Figure 4 illustrates pathological responses as a result of a clinical trial according to one embodiment of the present invention.

[0050] The detailed description of the present invention described below will be described with reference to specific drawings (if any) regarding specific embodiments in which the present invention may be practiced; however, the present invention is not limited thereto, but is defined only by the appended claims to the full scope equivalent to or equivalent to what the claims describe. It should be understood that the various embodiments / embodiments of the present invention, while different from each other, are not necessarily mutually exclusive. For example, specific shapes, structures, and characteristics described in this specification may be changed from one embodiment / embodiment to another, or multiple embodiments / embodiments may be combined, without departing from the spirit and scope of the present invention. Technical and scientific terms used herein have the same meaning as commonly used in the art to which the present invention belongs, unless otherwise defined. In case of conflict, the present specification, including its definitions, will control. The definitions of terms set forth in this specification will be applied for the purpose of interpreting this specification, and terms expressed in the singular will be construed to also refer to the plural (i.e., at least one) and vice versa, unless the context otherwise makes it inappropriate.

[0051] Methods for treating cancer or inhibiting cancer growth

[0052] In one aspect of the present invention, a method of treating a tumor or inhibiting tumor growth is provided, comprising the steps of selecting a patient with an upper gastrointestinal cancer (e.g., localized gastric cancer, esophageal cancer, or liver cancer) amenable to surgical resection, and administering to the patient a PD-L1 inhibitor (e.g., anti-PD-L1 antibody IMC-001 or a biological equivalent thereof) as neoadjuvant therapy prior to surgical resection of the tumor.

[0053] In one embodiment, the methods disclosed herein can further comprise administering to the patient a PD-L1 inhibitor (e.g., anti-PD-L1 antibody IMC-001 or a bioequivalent thereof) as an adjuvant therapy after completing surgery to treat an upper gastrointestinal cancer (e.g., gastric cancer, esophageal cancer, or liver cancer).

[0054] As used herein, "gastric cancer" refers to a general term for cancers (malignant tumors) arising in the stomach, including gastric adenocarcinoma, lymphoma, gastric submucosal tumor, and leiomyoma. Gastric cancer can arise in any part of the stomach and can spread throughout the stomach and to other organs, particularly the esophagus, lungs, and liver. In one embodiment, the gastric cancer is gastric adenocarcinoma. In one embodiment, the gastric cancer is resectable and recurrent. In one embodiment, the gastric cancer is metastatic.

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

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

[0057] In some embodiments, the patient is a candidate for surgery to resect a gastric, esophageal, or liver cancer tumor. In some embodiments, the patient suffers from gastric, esophageal, or liver cancer for which the surgical procedure is intended to be curative.

[0058] As used herein, the terms "treating," "treatment," or similar expressions mean alleviating or reducing the severity of at least one symptom or sign, temporarily or permanently eliminating a symptomatic factor, delaying or inhibiting tumor growth, reducing tumor cell burden or tumor burden, promoting tumor regression, causing tumor shrinkage, necrosis, and / or disappearance, preventing tumor recurrence, preventing or inhibiting metastasis, inhibiting metastatic tumor growth, eliminating the need for surgery, and / or increasing the survival time of a subject. In many embodiments, the terms "tumor," "lesion," "tumor lesion," "cancer," and "malignancy" are used interchangeably and refer to one or more malignant growths.

[0059] As used herein, the term "recurrence" refers to the frequent or recurrent diagnosis of gastric cancer, esophageal cancer, or liver cancer in a patient, or the frequent or recurrent occurrence of an individual tumor, such as a new tumor, which may represent a recurrence of a primary tumor and / or a previous tumor. In certain embodiments, administration of a PD-L1 inhibitor inhibits the recurrence of gastric cancer, esophageal cancer, or liver cancer in a patient.

[0060] As used herein, an "individual in need of a PD-L1 inhibitor" means a human or non-human mammal that exhibits one or more symptoms or signs of gastric, esophageal, or liver cancer and / or has been diagnosed with gastric, esophageal, or liver cancer and is in need of treatment therefor. In many embodiments, the terms "individual" and "patient" are used interchangeably. Such expressions include individuals with primary, established, or recurrent tumors (advanced malignancies). In certain embodiments, the expression includes human individuals who have been treated for and / or are in need of treatment for recurrent but non-metastatic gastric, esophageal, or liver cancer. In certain embodiments, the expression includes patients with solid tumors that have not been adequately controlled by, are resistant to, or are refractory to, prior therapy (e.g., surgery or treatment with an anticancer agent other than IMC-001 or a bioequivalent thereof). In certain embodiments, the expression includes individuals with gastric, esophageal, or liver cancer who are candidates for curative surgery.

[0061] In certain embodiments, the methods of the present invention are used to treat individuals with solid tumors. As used herein, the term "solid tumor" typically refers to an abnormal mass of tissue that does not contain cysts or areas of liquid. Solid tumors can be benign (non-cancerous) or malignant (cancerous). For the purposes of the present invention, the term "solid tumor" refers to a malignant solid tumor. This term encompasses several types of solid tumors, named according to the cell type that forms them, namely, sarcomas, carcinomas, and blastomas.

[0062] In certain embodiments, the methods of the present invention comprise administering a therapeutically effective amount of a PD-L1 inhibitor (e.g., the anti-PD-L1 antibody IMC-001 or a bioequivalent thereof) in combination with an additional therapeutic agent or therapy. The additional therapeutic agent or therapy may be used to enhance anti-tumor efficacy, reduce toxic effects of one or more therapies, and / or reduce the dosage of one or more therapies. In various embodiments, the additional therapeutic agent or therapy may include one or more of the following: an anti-viral therapy (e.g., cidofovir), photodynamic therapy, a PD-1 inhibitor (e.g., an antibody to PD-1 known in the art, such as nivolumab, pembrolizumab, etc.), a lymphocyte activation gene 3 (LAG3) inhibitor (e.g., an anti-LAG3 antibody), a cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) inhibitor (e.g., ipilimumab), a glucocorticoid-induced tumor necrosis factor receptor (GITR) agonist (e.g., an anti-GITR antibody), a T-cell immunoglobulin and mucin containing-3 (TIM3) inhibitor, a B- and T-lymphocyte attenuating factor (BTLA) inhibitor, a T-cell immunoreceptor with Ig and ITIM domains (TIGIT) inhibitor, CD38 inhibitors, CD47 inhibitors, antagonists of other T-cell co-inhibitors or ligands (e.g., antibodies to CD-28, 2B4, LY108, LAIR1, ICOS, CD160 or VISTA), CD20 inhibitors (e.g., anti-CD20 antibodies or bispecific CD3 / CD20 antibodies), indoleamine-2,3-dioxygenase (IDO) inhibitors, CD28 activators, vascular endothelial growth factor (VEGF) antagonists (e.g., "VEGFTrap" such as aflibercept or other VEGF-inhibiting fusion proteins listed in US 7087411), or anti-VEGF antibodies or antigen-binding fragments thereof (e.g.,bevacizumab or ranibizumab) or small molecule kinase inhibitors of VEGF receptors (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 to costimulatory receptors (e.g., agonists to CD28, 4-1BB, or OX40), antibodies to tumor-specific antigens (e.g., CA9, CA125, melanoma-associated antigen 3 (MAGE3), carcinoembryonic antigen (CEA), vimentin), tumor-M2-PK, prostate-specific antigen (PSA), mucin-1, MART-1, and CA19-9), vaccines (e.g., Bacillus Calmette-Guérin or cancer vaccines), adjuvants that increase antigen presentation (e.g., granulocyte-macrophage colony-stimulating factor), oncolytic viruses, cytotoxins, chemotherapeutic agents (e.g., pemetrexed, dacarbazine, temozolomide, cyclophosphamide, docetaxel, doxorubicin, daunorubicin, cisplatin, carboplatin, gemcitabine, methotrexate, mitoxantrone, oxaliplatin, paclitaxel, topotecan, irinotecan, vinorelbine, and vincristine), platinum-based chemotherapy (e.g., platinum-doublet chemotherapy), tyrosine kinase inhibitors (e.g., lenvatinib),Cytokines such as regorafenib and cabozantinib), IL-6R inhibitors, IL-4R inhibitors, IL-10 inhibitors, 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-targeted T cells), anti-inflammatory agents such as corticosteroids, non-steroidal anti-inflammatory drugs (NSAIDs), and dietary supplements such as antioxidants.

[0063] As used herein, the term "anti-viral therapy" refers to any substance, drug or therapy used to treat, prevent or ameliorate a viral infection in a host organism, including but not limited to zidovudine, lamivudine, abacavir, ribavirin, lopinavir, efavirenz, cobicistat, tenofovir, rilpivirine, analgesics, corticosteroids and combinations thereof. In the context of the present invention, chronic viral infections include infections caused by viruses, including but not limited to human immunodeficiency virus (HIV), hepatitis B virus (HBV) and hepatitis C virus (HCV).

[0064] In certain embodiments, administering a therapeutically effective amount of a PD-L1 inhibitor (e.g., anti-PD-L1 antibody IMC-001 or a bioequivalent thereof) to a subject suffering from gastric cancer, esophageal cancer, or liver cancer results in increased inhibition of tumor proliferation, e.g., greater tumor regression in the treated subject.

[0065] In certain embodiments, administration of a PD-L1 inhibitor achieves one or more of the following: (i) an improvement in tumor response rate, e.g., an improvement in overall response rate, complete response, or partial response, as compared to subjects treated with surgical resection alone or a non-treated subject; (ii) a delay in tumor growth and development, e.g., tumor growth can be delayed by about 3 days, >3 days, about 7 days, >7 days, >15 days, >1 month, >3 months, >6 months, >1 year, >2 years, or >3 years as compared to subjects treated with surgical resection alone or a non-treated subject; (iii) an increase in disease-free survival (DFS) until tumor recurrence or death as compared to subjects treated with surgical resection alone or a non-treated subject; and (iv) an improvement in overall response rate, complete response, or partial response, as compared to subjects treated with surgical resection alone or a non-treated subject. In certain embodiments, administering a therapeutically effective amount of a PD-L1 inhibitor (e.g., anti-PD-L1 antibody IMC-001 or a bioequivalent thereof) to a subject having gastric cancer, esophageal cancer, or liver cancer prevents tumor recurrence and / or increases survival of the subject, e.g., increases survival by >15 days, >1 month, >3 months, >6 months, >12 months, >18 months, >24 months, >36 months, or >48 months compared to a subject treated only with surgical resection or a non-treated subject.

[0066] In certain embodiments, administering a therapeutically effective amount of a PD-L1 inhibitor (e.g., the anti-PD-L1 antibody IMC-001 or a bioequivalent thereof) to a subject having gastric cancer, esophageal cancer, or liver cancer results in an increase in overall survival (OS) or progression-free survival (PFS) of the subject compared to a subject 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 a subject treated with surgical resection alone. In certain embodiments, 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 subjects treated with surgical resection alone.

[0067] PD-L1 inhibitors

[0068] The methods disclosed herein comprise administering a therapeutically effective amount of a PD-L1 inhibitor, wherein the PD-L1 inhibitor can be an anti-PD-L1 antibody, such as the fully human anti-PD-L1 antibody IMC-001 or a biological equivalent thereof. As used herein, the term "biological equivalent" refers to an anti-PD-L1 antibody or a PD-L1-binding protein or fragment thereof, as a pharmaceutical equivalent or alternative, that does not significantly differ from the IMC-001 antibody in rate and / or level of absorption when administered at the same molar dose as a single or multiple administrations under similar experimental conditions. In the context of the present invention, the term "biological equivalent" encompasses antibody-binding proteins that do not clinically differ significantly from the IMC-001 antibody in terms of safety, purity, and / or potency and that bind to PD-L1.

[0069] As used herein, the term "antibody" is intended to refer to immunoglobulin molecules (i.e., "full antibody molecules") composed of four polypeptide chains, two heavy (H) chains and two light (L) chains interconnected by disulfide bonds, as well as multimers thereof (e.g., IgM) or antigen-binding fragments thereof. Each heavy chain is comprised of a heavy chain variable region ("HCVR" or "VH") and a heavy chain constant region (composed of domains CH1, CH2, and CH3). Each light chain is composed of a light chain variable region ("LCVR" or "VL") and a light chain constant region (CL). The VH and VL regions can be further subdivided into hypervariable regions, called complementarity determining regions (CDRs), separated by more conserved regions, called framework regions (FRs). Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In certain embodiments, the FRs of an antibody (or antigen-binding fragment thereof) can be identical to a human germline sequence or can be naturally or artificially modified. An amino acid consensus sequence can be defined based on side-by-side analysis of two or more CDRs. As used herein, the term "antibody" also includes antigen-binding fragments of full-length antibody molecules.

[0070] As used herein, the terms "antigen-binding fragment" of an antibody, "antigen-binding portion" of an antibody, and the like, include naturally occurring, enzymatically obtainable, synthetic, 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, for example, proteolysis or recombinant genetic engineering techniques involving the manipulation and expression of DNA encoding antibody variable domains and, optionally, constant domains. Such DNA is known and / or readily available, for example, from commercial sources, DNA libraries (e.g., phage-antibody libraries), or can be synthesized. The DNA can be sequenced and manipulated chemically or using molecular biology techniques to, for example, arrange one or more variable and / or constant domains into a suitable configuration, introduce codons, construct cysteine ​​residues, or modify, add, or remove amino acids.

[0071] Non-limiting examples of antigen-binding fragments include: (i) a Fab fragment; (ii) an F(ab')2 fragment; (iii) an Fd fragment; (iv) an Fv fragment; (v) a single-chain Fv (scFv) molecule; (vi) a dAb fragment; and (vii) a minimal recognition unit composed of amino acid residues that mimics the hypervariable region of an antibody (e.g., an isolated complementarity determining region (CDR) such as a CDR3 peptide), or a restricted FR3-CDR3-FR4 peptide. Other engineered molecules, such as domain-specific antibodies, single domain antibodies, domain-deleted antibodies, chimeric antibodies, CDR-grafted antibodies, diabodies, triabodies, tetrabodies, minibodies, nanobodies (e.g., monovalent nanobodies, bivalent nanobodies, etc.), small modular immunopharmaceuticals (SMIPs), and shark variable IgNAR domains, are also encompassed by the expression "antigen-binding fragment" as used herein.

[0072] An antigen-binding fragment of an antibody will typically comprise at least one variable domain. The variable domain may be of any size or amino acid composition and will generally comprise at least one CDR adjacent to or in-frame with one or more framework sequences. In an antigen-binding fragment having a VH domain associated with a VL domain, the VH and VL domains may be arranged in any suitable arrangement relative to each other. For example, the variable domains may be dimeric and may contain VH-VH, VH-VL, or VL-VL dimers. Alternatively, the antigen-binding fragment of an antibody may contain monomeric VH or VL domains.

[0073] In certain embodiments, an antigen-binding fragment of an antibody may contain at least one variable domain covalently linked to at least one constant domain. Non-limiting exemplary structures for variable and constant domains that may be found in an antigen-binding fragment of an antibody of the 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 constant domains, including any of the exemplary structures described above, the variable and constant domains may be directly connected to each other or may be connected to each other by the full length or part of a hinge or a linker region. The hinge region may be composed of at least two (e.g., 5, 10, 15, 20, 40, 60, or more) amino acids that form a single polypeptide molecule through a flexible or semi-flexible linkage between adjacent variable and / or constant domains. Furthermore, antigen-binding fragments of antibodies of the invention may comprise homo-dimers or hetero-dimers (or other multimers) of any of the variable and constant domain arrangements described above, non-covalently associated with each other and / or with one or more monomeric VH or VL domains (e.g., by disulfide bond(s)).

[0074] The antibodies utilized in the methods of the present invention may be human antibodies. As used herein, the term "human antibody" refers to an antibody having variable domains and constant regions derived from human germline immunoglobulin sequences. Human antibodies of the present invention may, however, include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced in vitro by random or site-directed mutagenesis or in vivo by somatic mutation), for example, in the CDRs, particularly CDR3. However, as used herein, the term "human antibody" is not intended to encompass antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, are grafted onto human framework sequences. In one embodiment, all variable and constant domains may be derived from human immunoglobulin sequences (a fully human antibody).

[0075] The antibodies utilized in the methods disclosed herein may be recombinant human antibodies. As used herein, the term "recombinant human antibody" encompasses any human antibody that is prepared, expressed, constructed, or isolated by recombinant means, such as antibodies expressed using a recombinant expression vector transfected into a host cell (described further below), antibodies isolated from a recombinant, combinatorial human antibody library (described further below), antibodies isolated from an animal (e.g., a mouse) that is transgenic for human immunoglobulin genes (see, e.g., Taylor et al. (1992) Nucl. Acids Res. 20:6287-6295), or antibodies prepared, expressed, constructed, or isolated by any other means that involves splicing of human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. However, in certain embodiments, such recombinant human antibodies are mutagenized in vitro (or, when using animals transgenic for human Ig sequences, somatically mutagenized in vivo), and thus the amino acid sequences of the VH and VL regions of the recombinant antibodies are sequences that are derived from and related to human germline VH and VL sequences, but which may not be natively present in the human antibody germline repertoire in vivo.

[0076] In some embodiments, the PD-L1 inhibitor is an anti-PD-L1 antibody (e.g., IMC-001) comprising three heavy chain complementary determining regions (HCDRs) of a heavy chain variable region (HCVR) comprising or consisting of the amino acid sequence of SEQ ID NO: 1 and three light chain complementary determining regions (LCDRs) of a light chain variable region (LCVR) comprising or consisting of the amino acid sequence of SEQ ID NO: 2. In some embodiments, the anti-PD-L1 antibody comprises three HCDRs (HCDR1, HCDR2, and HCDR3) and three LCDRs (LCDR1, LCDR2, and LCDR3), wherein HCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 5; HCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 6; HCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 7; LCDR1 comprises or consists of the amino acid sequence of SEQ ID NO: 8; LCDR2 comprises or consists of the amino acid sequence of SEQ ID NO: 9; and LCDR3 comprises or consists of the amino acid sequence of SEQ ID NO: 10. In certain embodiments, the anti-PD-L1 antibody comprises an HCVR comprising or consisting of SEQ ID NO: 1 and an LCVR comprising or consisting of SEQ ID NO: 2. In certain embodiments, the anti-PD-L1 antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises or consists of the amino acid sequence of SEQ ID NO: 3, and the light chain comprises or consists of the amino acid sequence of SEQ ID NO: 4. 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 herein as "IMC-001."

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

[0078] In some embodiments, the sequence identity may be to a region excluding the CDR regions (e.g., a framework sequence). In some embodiments, a biological equivalent of the IMC-001 antibody is an anti-PD-L1 antibody comprising an HCVR having at least 90%, at least 95%, at least 97%, or at least 98% sequence identity to a region excluding the HCDR1, HCDR2, and HCDR3 regions of SEQ ID NO: 1. In some embodiments, a biological equivalent of the IMC-001 antibody is an anti-PD-L1 antibody comprising an LCVR having at least 90%, at least 95%, at least 97%, or at least 98% sequence identity to a region excluding the LCDR1, LCDR2, and LCDR3 regions of SEQ ID NO: 2. In some embodiments, a biological equivalent of IMC-001 is an anti-PD-L1 antibody comprising an HCVR having at least 90%, at least 95%, at least 97%, or at least 98% sequence identity to a region excluding the HCDR1, HCDR2, and HCDR3 regions of SEQ ID NO: 1, and an LCVR having at least 90%, at least 95%, at least 97%, or at least 98% sequence identity to a region excluding the LCDR1, LCDR2, and LCDR3 regions of SEQ ID NO: 2. In some embodiments, the biological equivalent of the IMC-001 antibody is an anti-PD-L1 antibody comprising an LCVR having at least 90%, at least 95%, at least 97%, or at least 98% sequence identity to a region excluding the LCDR1, LCDR2, and LCDR3 regions of SEQ ID NO: 2, and wherein amino acids 3 and 5 from N-terminus to C-terminus of SEQ ID NO: 2 are identical to amino acids 3 and 5 of SEQ ID NO: 2.

[0079] In some embodiments, the IMC-001 antibody or a biological equivalent thereof can be produced, particularly in CHO cells, without involving light chain fragmentation by having a light chain amino acid sequence as described above.

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

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

[0082] In one embodiment, the anti-PD-L1 antibody binds to a PD-L1 epitope at a density of 10 -6A 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 a PD-L1 epitope is provided, wherein the antibody comprises a heavy chain variable domain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence set forth in SEQ ID NO: 1, and a light chain variable domain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence set forth in SEQ ID NO: 2. In one embodiment, the invention provides a fully human antibody of the IgG class (e.g., IgG1 or IgG4) that binds to a PD-L1 epitope, wherein the antibody comprises a heavy chain variable domain comprising a CDR1 domain, a CDR2 domain and a CDR3 domain as set forth 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 comprising a CDR1 domain, a CDR2 domain and a CDR3 domain as set forth in the amino acid sequences of SEQ ID NO: 8, SEQ ID NO: 9 and SEQ ID NO: 10, respectively.

[0083] For more specific information regarding the anti-PD-L1 antibody of the present invention, please refer to International Publication No. WO 2017 / 132562 A1. The IMC-001 antibody comprises 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 in the above document, the entire disclosure of which is incorporated herein by reference.

[0084] Manufacturing of PD-L1 inhibitors

[0085] Antigen binding proteins can be produced by any of a number of conventional techniques. In one embodiment, the present invention provides a monoclonal antibody that binds PD-L1. The monoclonal antibody can be purified, for example, from cells that naturally express it (e.g., the antibody can be purified from a hybridoma producing it), or produced in a recombinant expression system using any technique known in the art. See, e.g., Monoclonal Antibodies, Hybridomas: A New Dimension in Biological Analyses, Kennet et al. (eds.), Plenum Press, New York (1980); and Antibodies: A Laboratory Manual, Harlow and Land (eds.), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, (1988).

[0086] Monoclonal antibodies can be produced using any technique known in the art, for example, by immortalizing spleen cells harvested from a transgenic animal after completion of the immunization schedule. The spleen cells can be immortalized using any technique known in the art, for example, by fusing them with myeloma cells to produce hybridomas. The myeloma cells for use in the hybridoma-producing fusion procedure are preferably non-antibody-producing myeloma cells, exhibit high fusion efficiency, and have enzyme deficiencies that render them incapable of growth in specific selective media that support the growth of only the desired fused cells (hybridomas). Examples of cell lines suitable for use in mouse fusions 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 fusions include R210.RCY3, Y3-Ag 1.2.3, IR983F, and 48210. Other cell lines useful for cell fusions include U-266, GM1500-GRG2, LICR-LON-HMy2, and UC729-6.

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

[0088] For recombinant production of anti-PD-L1 antibodies, nucleic acids encoding the antibody 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 conventional procedures (e.g., using oligonucleotide probes capable of specifically binding to genes encoding the heavy and light chains of the antibody).

[0089] Nucleic acids encoding the anti-PD-L1 antibodies (or fragments) disclosed herein can be chemically synthesized. Codon usage can be selected to improve expression in cells. This codon usage will depend on the cell type selected. Specific codon usage patterns have been developed for E. coli and other bacteria, as well as mammalian cells, plant cells, yeast cells, and insect cells. See, e.g., 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; and Sharp et al. Yeast. See [1991 7(7):657-78].

[0090] General techniques for nucleic acid manipulation are described, for example, in Sambrook et al., Molecular Cloning: A Laboratory Manual, Vols. 1-3, Cold Spring Harbor Laboratory Press, 2nd ed., 1989, or F. Ausubel et al., Current Protocols in Molecular Biology (Green Publishing and Wiley-Interscience: New York, 1987), and as periodically updated, which are incorporated herein by reference. The DNA encoding the polypeptide is operably linked to appropriate transcriptional or translational control elements derived from mammalian, viral, or insect genes. Such control elements include a transcriptional promoter, an optional operator sequence for regulating transcription, a sequence encoding a suitable mRNA ribosome binding site, and sequences regulating the termination of transcription and translation. Typically, a selection gene is additionally included to facilitate the ability to replicate in the host, conferred by the origin of replication, and recognition of the transformant.

[0091] Additionally, the recombinant DNA may contain any type of protein tag sequence that may be useful for purifying the protein. Examples of protein tags include, but are not limited to, histidine tags, FLAG tags, myc tags, HA tags, or GST tags. Suitable cloning and expression vectors for use with bacterial, fungal, yeast, and mammalian cell hosts can be found in Cloning Vectors: A Laboratory Manual (Elsevier, NY, 1985).

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

[0093] Any expression system known in the art can be used to produce recombinant polypeptides (e.g., recombinant antibodies) of the present invention. Typically, a host cell is transformed with a recombinant expression vector containing DNA encoding the desired polypeptide. Host cells that can be used include prokaryotes, yeast, or higher eukaryotic cells. Prokaryotes include gram-negative or gram-positive organisms, such as E. coli or bacilli. Higher eukaryotic cells include insect cells and established cell lines of mammalian origin. Suitable host cells for cloning or expressing antibody-encoding vectors include the prokaryotic or eukaryotic cells described herein. For example, antibodies can be produced in bacteria, particularly when glycosylation and Fc effector function are not required. For the expression of antibody fragments and polypeptides in bacteria, see, e.g., U.S. Patent Nos. 5,648,237, 5,789,199, and 5,840,523. (See also Charlton, Methods in Molecular Biology, Vol. 248 (BKC Lo, ed., Humana Press, Totowa, NJ, (2003), pp. 245-254) which describes the expression of antibody fragments in E. coli. After expression, the antibody can be isolated from the bacterial cell paste in the soluble fraction and further purified. Higher eukaryotic cells include insect cells and established cell lines of mammalian origin. 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 lines and the like described in McMahan et al. al., 1991, EMBO J.10: 2821], including the CV1 / EBNA cell line (ATCC CCL 70), derived from the African green monkey kidney cell line CV1. Suitable cloning and expression vectors for use with bacterial, fungal, yeast, and mammalian cell hosts are described by Pouwels et al. [Cloning Vectors: A Laboratory Manual, (Elsevier, NY, 1985)].

[0094] In certain embodiments, vertebrate cells may be used as hosts to express anti-PD-L1 antibodies or fragments thereof. For example, mammalian cell lines adapted to grow in suspension may be useful. Other examples of useful mammalian host cell lines include monkey kidney CV1 cells transformed by SV40 (COS-7); human embryonic kidney cells (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); Dog kidney cells (MDCK; buffalo rat hepatocytes (BRL 3A); human lung cells (W138); human hepatocytes (Hep G2); mouse mammary tumor (MMT 060562); TRI cells, as described in, e.g., Mather et al., Annals NY Acad. Sci. 383:44-68 (1982); MRC 5 cells; and FS4 cells. 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))—which contain the DHFR; and myeloma cell lines, such as Y0, NS0, and Sp2 / 0. For a review of specific mammalian host cell lines suitable for antibody production, see, e.g., 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 (ATCC CRL 1651) of monkey kidney cells (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 lines, and the CV1 / EBNA cell line (ATCC CCL 70) derived from the African green monkey kidney cell line CV1 as described by McMahan et al., 1991, EMBO J. 10: 2821.

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

[0096] Suitable host cells for the expression of glycosylated antibodies are derived from multicellular organisms (both invertebrates and vertebrates). Examples of invertebrate cells include plant and insect cells. Various baculovirus strains have been identified that can be used in conjunction with insect cells, particularly for transfection of Spodoptera frugiperda cells.

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

[0098] Transformed cells can be cultured under conditions that promote expression of the polypeptide, and the polypeptide can be recovered using conventional protein purification procedures. One such purification procedure involves, for example, the use of affinity chromatography on a matrix with all or a portion (e.g., the extracellular domain) of PD-L1 bound to the matrix. Polypeptides contemplated for use herein include substantially homogeneous recombinant mammalian anti-PD-L1 antibody polypeptides that are substantially free of endogenous contaminants.

[0099] Accordingly, 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 an anti-PD-L1 antibody described herein is provided. Such nucleic acid may encode an amino acid sequence comprising a VL of the antibody and / or an amino acid sequence comprising a VH of the antibody (e.g., a light chain and / or a heavy chain of the antibody). In a further embodiment, one or more vectors (e.g., expression vectors) comprising such nucleic acids are provided. In a further embodiment, a host cell comprising such nucleic acids is provided. In one such embodiment, the host cell comprises (e.g., is transformed with): (1) a vector comprising a nucleic acid encoding an amino acid sequence comprising a VL of the antibody and an amino acid sequence comprising a VH of the antibody, or (2) a first vector comprising a nucleic acid encoding an amino acid sequence comprising a VL of the antibody and a second vector comprising a nucleic acid encoding an amino acid sequence comprising a VH of the antibody. In one embodiment, the host cell is a eukaryote, such as a Chinese hamster ovary (CHO) cell or a lymphoid cell (e.g., Y0, NS0, Sp20 cell). In one embodiment, a method of producing an anti-PD-L1 antibody is provided, wherein the method comprises culturing a host cell comprising a nucleic acid encoding an antibody as provided above under conditions suitable for expression of the antibody, and optionally recovering the antibody from the host cell (or host cell culture medium).

[0100] The proteins disclosed herein can also be produced using a cellular translation system. For this purpose, the nucleic acid encoding the polypeptide must be modified to allow for in vitro transcription to produce mRNA and for cell-free translation of the mRNA in the specific cell-free system being utilized (eukaryotic, e.g., mammalian or yeast cell-free translation systems, or prokaryotic, e.g., bacterial cell-free translation systems).

[0101] Additionally, PD-L1-binding polypeptides can be prepared by chemical synthesis (e.g., by the method described in Solid Phase Peptide Synthesis, 2nd ed., 1984, The Pierce Chemical Co., Rockford, III.). Modifications to the protein can also be made by chemical synthesis.

[0102] The polypeptides of the present invention can be purified by any separation / purification method for proteins generally known in the art 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, reversed phase chromatography, gel filtration, gel permeation chromatography, affinity chromatography, electroporation, countercurrent distribution, or any combination thereof. After purification, the polypeptides can be exchanged into various buffers and / or concentrated by any of a variety of methods known in the art, including, but not limited to, filtration and dialysis.

[0103] The purified polypeptide is preferably at least 85% pure, more preferably at least 95% pure, and most preferably at least 98% pure. Regardless of the exact degree of purity, the polypeptide is sufficiently pure for use as a pharmaceutical product. An antigen binding protein (e.g., an antibody, antibody fragment, antibody derivative, antibody mutein, or antibody variant) is a polypeptide that binds to PD-L1 (preferably human PD-L1). An antigen binding protein includes an antigen binding protein that inhibits the biological activity of PD-L1.

[0104] Antigen binding proteins can be produced by any of a number of known techniques and screened for desired properties. Particular techniques involve isolating a nucleic acid encoding a polypeptide chain (or portion thereof) of an antigen binding protein of interest (e.g., an anti-PD-L1 antibody) and manipulating the nucleic acid using recombinant DNA techniques. The nucleic acid may be fused to another nucleic acid of interest or modified (e.g., using mutagenesis or other conventional techniques) to, for example, add, delete, or substitute one or more amino acid residues.

[0105] Single-chain antibodies can be formed by linking fragments of the heavy and light chain variable domains (Fv regions) via an amino acid bridge (a short peptide linker) to form a single polypeptide chain. The single-chain Fv (scFv) has been prepared by fusing DNA encoding a peptide linker between DNAs encoding two variable domain polypeptides (VL and VH). The resulting polypeptides can self-fold 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 multimeric scFvs 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; Bird, 1988, Science 242:423; Huston et al., 1988, Proc. Natl. Acad. Sci. USA 85:5879; Ward et al., 1989, Nature 334:544; de Graaf et al., 2002, Methods Mol. Biol. 178:379-87).

[0106] Techniques for deriving antibodies of different subclasses or isotypes from an antibody of interest, called subclass switching, are known. Thus, for example, an IgG antibody can be derived from an IgM antibody, and conversely, an IgM antibody can be derived from an IgG antibody. This technique allows for the production of new antibodies that retain the antigen-binding properties of a given antibody (the parent antibody) but also exhibit biological properties associated with a different antibody isotype or subclass than those of the parent antibody. Recombinant DNA techniques can be utilized. Cloned DNA encoding a specific antibody polypeptide, such as DNA encoding the constant domain of an antibody of the desired isotype, can be used in this procedure (Lantto et al., 2002, Methods Mol. Biol. 178:303-16). Additionally, if IgG4 is preferred, it may also be desirable to introduce a point mutation (CPSCP->CPPCP) in the hinge region to mitigate the tendency to form inter-H chain disulfide bonds, which can cause heterogeneity in IgG4 antibodies (Bloom et al., 1997, Protein Science 6:407).

[0107] Pharmaceutical compositions and administration

[0108] Another aspect of the present invention provides a pharmaceutical composition comprising a PD-L1 inhibitor as a neoadjuvant therapy disclosed herein. Such a pharmaceutical composition may be formulated with suitable pharmaceutically acceptable carriers, excipients, buffers, and other materials that provide suitable transport, delivery, tolerability, etc. Many suitable formulations can be found in formularies known to all pharmacists: Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA. These formulations include, for example, powders, pastes, ointments, jellies, waxes, oils, lipids, (LIPOFECTIN)TM Examples include lipid (cationic or anionic)-containing vesicles, DNA conjugates, anhydrous absorption pastes, oil-in-water and water-in-oil emulsions, emulsified carbowaxes (polyethylene glycols 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).

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

[0110] Various delivery systems are known, such as encapsulation in liposomes, microparticles, microcapsules, recombinant cells capable of expressing the mutant virus, and receptor-mediated endocytosis (see, e.g., Wu et al. (1987) J. Biol. Chem. 262:4429-4432), which can be used to administer the pharmaceutical compositions of the present invention. Routes of introduction include, but are not limited to, intradermal, transdermal, intramuscular, intravenous, subcutaneous, intranasal, epidural, and oral routes. The compositions may be administered by any convenient route, for example, by infusion 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 may also be delivered in the form of vesicles, particularly liposomes (see, e.g., Langer (1990) Science 249:1527-1533).

[0111] The use of nanoparticles to deliver PD-L1 inhibitors of the present invention is also contemplated herein. Antibody-conjugated nanoparticles can be utilized 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, et 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 for target cell delivery. Nanoparticles for drug delivery are also described, for example, in US Pat. No. 8,257,740 or US Pat. No. 8,246,995.

[0112] In some situations, the pharmaceutical composition may be delivered in the form of a controlled release system. In one embodiment, a pump may be used. In another embodiment, a polymeric material may be used. In yet another embodiment, the controlled release system is positioned near the target of the composition, so that only a fraction of the systemic dose may be required.

[0113] Injectable formulations may include dosage forms for intravenous, subcutaneous, intracranial, and intramuscular injection, as well as drip infusion. These injectable formulations may be prepared by publicly known methods. In one embodiment, the injectable formulation may be a histidine preparation solution.

[0114] The pharmaceutical composition of the present invention can be delivered subcutaneously or intravenously using a standard needle and syringe. Furthermore, for subcutaneous delivery, a pen delivery device is readily useful for delivering the pharmaceutical composition of the present invention. Such pen delivery devices may be reusable or disposable. Reusable pen delivery devices typically utilize a replaceable cartridge containing the pharmaceutical composition. Once all of the pharmaceutical composition in the cartridge has been dispensed 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 a replaceable cartridge. Instead, disposable pen delivery devices have a reservoir pre-filled with the pharmaceutical composition. Once the reservoir is empty, the empty device is discarded.

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

[0116] 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 a biological equivalent thereof) and a pharmaceutically acceptable carrier. Non-limiting examples of pharmaceutical compositions comprising anti-PD-L1 antibodies provided herein that may be used in the context of the present invention are disclosed in US 2019 / 0040137.

[0117] Also, another aspect of the present invention provides a kit comprising a PD-L1 inhibitor (e.g., IMC-001 or a biological equivalent thereof) for therapeutic use as described herein. The kit typically includes a label indicating the intended use of the contents of the kit and instructions for use. As used herein, the term "label" includes any written or recorded material provided on, in, or with the kit, or otherwise attached to the kit. In one embodiment, a kit is provided for treating a patient suffering from an upper gastrointestinal cancer (e.g., gastric cancer, esophageal cancer, or liver cancer), the kit comprising: (a) a therapeutically effective amount of a PD-L1 inhibitor (e.g., IMC-001 or a biological equivalent thereof); and (b) instructions for using the PD-L1 inhibitor in any of the methods disclosed herein.

[0118] Dosage and dosage

[0119] In certain embodiments, the methods of the invention can comprise administering to the tumor of the subject a therapeutically effective amount of a PD-L1 inhibitor (e.g., an IMC-001 antibody or a biological equivalent thereof) one or more times, for example, 1 time, 2 times, 3 times, 4 times, 5 times, 6 times, 7 times, 8 times, 9 times, or 10 times. For example, a therapeutic dosing regimen can include one or more administrations of a PD-L1 inhibitor given 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.

[0120] In certain embodiments, the PD-L1 inhibitor is administered in one or more treatment cycles, for example, one, two, three, four, five, six, seven, eight, nine, or ten treatment cycles. Methods according to this aspect include administering to a subject in need thereof one or more neoadjuvant cycles, and optionally one or more adjuvant cycles, each treatment cycle comprising administering the PD-L1 inhibitor (e.g., IMC-001 or a bioequivalent thereof) one, two, three, four, five, six, seven, eight, nine, ten, or more times.

[0121] In one embodiment, the neoadjuvant PD-L1 inhibitor is administered 1, 2, 3, 4, 5, or 6 times prior to surgical resection of the tumor. In one embodiment, the neoadjuvant PD-L1 inhibitor is administered 2 to 6 times prior to surgical resection of the tumor. In one embodiment, the neoadjuvant PD-L1 inhibitor is administered a total of 2 times (2 treatment cycles) prior to surgical resection of the tumor. In one embodiment, the neoadjuvant PD-L1 inhibitor is administered 2 to 6 times (6 treatment cycles) prior to surgical resection of the tumor. In one embodiment, the neoadjuvant PD-L1 inhibitor is administered 2 to 6 times (2 treatment cycles) prior to surgical resection of the tumor. In one embodiment, the neoadjuvant PD-L1 inhibitor is administered about once every week, about every 2 weeks, or about every 3 weeks. In one embodiment, the PD-L1 inhibitor as neoadjuvant therapy is administered at intervals of 10 to 18 days, preferably at intervals of 14 days.

[0122] In certain embodiments, each dose of the PD-L1 inhibitor comprises 0.1, 1, 0.3, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 mg / kg of body weight of the patient. 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 twice, 10 to 18 days apart, at a dose of 20 mg / kg prior to surgical resection of the tumor (2 cycles of administration).

[0123] The amount of a PD-L1 inhibitor (e.g., IMC-001 or a bioequivalent thereof) administered to a subject according to the methods disclosed herein is generally a therapeutically effective amount. As used herein, the term "therapeutically effective amount" means an amount of a PD-L1 inhibitor administered as neoadjuvant therapy prior to surgery planned to treat gastric cancer, esophageal cancer, or liver cancer that achieves one or more of the following: (a) inhibition of tumor growth or an increase in tumor necrosis, tumor shrinkage, and / or tumor disappearance, as compared to a subject treated with surgical resection alone or a non-treated subject, respectively: (b) a decrease in symptoms or signs of cancer, e.g., the severity or duration of tumor lesions; (c) a delay in tumor growth and progression; (d) an inhibition of tumor metastasis; (e) a prevention of recurrence of tumor growth; and / or (f) an increase in the survival of a subject suffering from cancer.

[0124] In certain embodiments, a therapeutically effective amount of a PD-L1 inhibitor (e.g., IMC-001 or a bioequivalent thereof) can be from about 0.05 mg to about 1000 mg, from about 1 mg to about 800 mg, from about 5 mg to about 600 mg, from about 10 mg to about 550 mg, from about 50 mg to about 400 mg, from about 75 mg to about 350 mg, or from about 100 mg to about 300 mg of antibody. For example, in various embodiments, the amount of PD-L1 inhibitor is about 0.05 mg, about 0.1 mg, about 1.0 mg, about 1.5 mg, about 2.0 mg, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 200 mg, about 210 mg, about 220 mg, about 230 mg, about 240 mg, about 250 mg, about 260 mg, about 270 mg, about 280 mg, about 290 mg, about 300 mg, about 310 mg, about 320 mg, about 330 mg, about 340 mg, about 350 mg, about 360 mg, about 370 mg, about 380 mg, about 390 mg, about 400 mg, about 410 mg, about 420 mg, about 430 mg, about 440 mg, about 450 mg, about 460 mg, about 470 mg, about 480 mg, about 490 mg, about 500 mg, about 510 mg, about 520 mg, about 530 mg, about 540 mg, about 550 mg, about 560 mg, about 570 mg, about 580 mg, about 590 mg, about 600 mg, about 610 mg, about 620 mg, about 630 mg, about 640 mg, about 650 mg, about 660 mg, about 670 mg, about 680 mg,about 690 mg, about 700 mg, about 710 mg, about 720 mg, about 730 mg, about 740 mg, about 750 mg, about 760 mg, about 770 mg, about 780 mg, about 790 mg, about 800 mg, about 810 mg, about 820 mg, about 830 mg, about 840 mg, about 850 mg, about 860 mg, about 870 mg, about 880 mg, about 890 mg, about 900 mg, about 910 mg, about 920 mg, about 930 mg, about 940 mg, about 950 mg, about 960 mg, about 970 mg, about 980 mg, about 990 mg, or about 1000 mg.

[0125] The amount of PD-L1 inhibitor (e.g., IMC-001 or a bioequivalent thereof) contained in an individual dose may be expressed as 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 at a dose of about 0.0001 to about 100 mg / kg of body weight of the subject. In certain embodiments, the anti-PD-L1 antibody can be administered to the subject at a dose of about 0.1 mg / kg to about 30 mg / kg of body weight of the subject. In certain embodiments, the methods of the invention comprise administering a PD-L1 inhibitor (e.g., anti-PD-L1 antibody IMC-001 or a bioequivalent thereof) at a dose of about 1 mg / kg to 30 mg / kg, 5 mg / kg to 30 mg / kg, 10 mg / kg to 30 mg / kg, 15 mg / kg, or 20 mg / kg of body weight of the subject.

[0126] In certain embodiments, the individual dose amount of the PD-L1 inhibitor (e.g., IMC-001 or a bioequivalent thereof) administered to the patient may be less than a therapeutically effective amount, i.e., a subtherapeutic dose. For example, if the therapeutically effective amount of the PD-L1 inhibitor comprises 20 mg / kg, a subtherapeutic dose comprises 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 a PD-L1 inhibitor that does not, by itself, elicit a therapeutic effect. However, in certain embodiments, multiple administrations of a PD-L1 inhibitor at subtherapeutic doses are performed to collectively achieve a therapeutic effect in the subject.

[0127] In certain embodiments, each dose comprises 0.1 to 30 mg / kg (e.g., 5 mg / kg, 10 mg / kg, 15 mg / kg, 20 mg / kg, or 25 mg / kg) of the PD-L1 inhibitor based on the patient's body weight. In some other embodiments, each dose comprises 5 to 600 mg of the PD-L1 inhibitor, e.g., 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.

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

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

[0130] In certain embodiments, the anti-PD-L1 antibody is administered twice at a dose of 20 mg / kg, spaced 10 to 18 days apart or 2 weeks apart, prior to surgical resection of the tumor in the patient, and the surgical resection of the tumor is performed 11 to 42 days after the last dose of the anti-PD-L1 antibody.

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

[0132] Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are intended only to illustrate the present invention and the scope of the present invention is not limited to these examples.

[0133] Example

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

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

[0136] 구분서열서열번호중쇄 가변도메인QMQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAYSWVRQAPGQGLEWMGGIIPSFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGPIVATITPLDYWGQGTLVTVSS1경쇄 가변도메인SYVLTQPPSVSVAPGKTATIACGGENIGRKTVHWYQQKPGQAPVLVIYYDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCLVWDSSSDHRIFGGGTKLTVL2중쇄MEWSWVFLFFLSVTTGVHSQMQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAYSWVRQAPGQGLEWMGGIIPSFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGPIVATITPLDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK3경쇄MSVPTQVLGLLLLWLTDARCSYVLTQPPSVSVAPGKTATIACGGENIGRKTVHWYQQKPGQAPVLVIYYDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCLVWDSSSDHRIFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS4중쇄 가변도메인 CDR1SYAYS5중쇄 가변도메인CDR2GIIPSFGTANYAQKFQG6Heavy chain variable domain CDR3GPIVATITPLDY7Light chain variable domain CDR1GGENIGRKTVH8Light chain variable domain CDR2YDSDRPS9Light chain variable domain CDR3LVWDSSSDHRI10

[0137]

[0138] IMC-001 is a mutant of the wild-type parent antibody H6B1L, H6B1L-EM, produced in CHO cells. For further details, see International Publication No. WO 2017 / 132562 A1.

[0139] 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.

[0140] Research Purpose

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

[0142] reason

[0143] Neoadjuvant immunotherapy has potential biological advantages, including a less immunosuppressed TME, lower disease burden, and anti-tumor immune priming in the presence of the primary tumor. Gastrointestinal cancers, including gastroesophageal cancer and hepatocellular carcinoma, are cancer types in which immune checkpoint inhibitors (ICIs) have shown a survival benefit over conventional therapy. IMC-001, a fully human anti-PD-L1 recombinant monoclonal antibody that maintains Fc effectors to stimulate ADCC, demonstrated a favorable safety profile in a phase 1 study (n = 15) at doses up to 20 mg / kg iv every 2 weeks and showed promising preliminary efficacy in patients with previously treated solid tumors (1 partial response and 4 stable disease) (Keam B, et al, Invest New Drugs 2021;39:1624-32).

[0144] Study endpoint

[0145] The primary endpoint of this study is to evaluate the major pathologic response rate in the surgical tissue after preoperative administration of the neoadjuvant immune checkpoint inhibitor IMC-001. Secondary endpoints include the feasibility and safety of preoperative neoadjuvant IMC-001 in delaying scheduled surgery, the R0 resection rate, the clinical tumor response rate according to RECIST v1.1, the clinical disease control rate according to RECIST v1.1, the progression-free survival (PFS), the relapse-free survival (RPS), the overall survival (OS), and the rates of cancer progression / recurrence and the patterns of cancer progression / recurrence. Meanwhile, the exploratory objective of this study is to secure subject samples (tissue, blood, stool) before and after administration of the immune checkpoint inhibitor IMC-001, establish a biomarker cohort, and conduct immune profiling and genome analysis to discover biomarkers predicting therapeutic efficacy.

[0146] Experimental design

[0147] Subjects who met the inclusion / exclusion criteria were assigned to respective cohorts for gastric cancer, esophageal cancer, and hepatocellular carcinoma. Each cancer cohort received two cycles of IMC-001 (each cycle consisting of two weeks) as neoadjuvant therapy, followed by curative surgery and follow-up. After curative surgery, adjuvant therapy was administered according to standard clinical guidelines, as appropriate, depending on the stage of the disease.

[0148] Patients with previously untreated resectable localized gastric, esophageal, or hepatocellular carcinoma were enrolled in this trial, and a total of 48 patients (16 per cancer cohort) were enrolled in this trial.

[0149] neoadjuvant therapy

[0150] In the absence of unacceptable toxicity or clinical progression, or withdrawal of consent, IMC-001 at 20 mg / kg was administered twice every 2 weeks as neoadjuvant therapy before surgical resection.

[0151] Biomarker sample collection

[0152] Tumor tissue, blood, and stool were collected before IMC-001 administration, blood and stool were collected after cycle 1 and cycle 2 administration, and surgical tumor tissue, blood, and stool were collected after surgery.

[0153] Follow-up observation

[0154] Blood, stool, and tumor tissue (only in case of recurrence) were collected and observed every 3 months for the first 2 years from the date of surgery, every 6 months for the next 3 years, and in case of cancer recurrence.

[0155] clinical trial subjects

[0156] A total of 48 patients (16 per cancer cohort) with gastric adenocarcinoma, esophageal squamous cell carcinoma, and hepatocellular carcinoma amenable to radical surgical resection were enrolled in this trial.

[0157] Selection criteria

[0158] Selection criteria for diseases

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

[0160] 2) Gastric adenocarcinoma, esophageal squamous cell carcinoma, or hepatocellular carcinoma expected to be amenable to radical resection

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

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

[0163] - Hepatocellular carcinoma: A single hepatocellular carcinoma confined to the liver, or three or fewer hepatocellular carcinomas confined to the liver. Invasion of the portal vein, hepatic vein, or biliary tract is possible, but there must be no invasion of the main portal trunk.

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

[0165] - Absolute neutrophil count ≥ 1,000 / μL

[0166] - Platelet count ≥75,000 / μL

[0167] - Serum total bilirubin ≤1.5 x upper limit of normal (ULN) (total bilirubin ≤3.0 x ULN for subjects with Gilbert syndrome)

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

[0169] - 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 below according to gender, and if the value is less than 50 mL / min, a test is conducted by collecting urine for 24 hours, and registration is possible if the creatinine clearance is 50 mL / min or higher)

[0170] [Creatinine clearance in men]

[0171]

[0172] [Creanine clearance in women]

[0173]

[0174] - Urine protein-creatinine ratio (UPC) ≤1 (However, if UPC >1, registration is possible if 24-hour urine protein is collected and less than 2g is collected)

[0175] - In addition, in the case of hepatocellular carcinoma, sufficient liver function must be confirmed with Child-Pugh grade A (see Table 4) and encephalopathy grade 0 (see Table 3) (see Table 2).

[0176]

[0177]

[0178]

[0179] 4) Measurable lesion or evaluable lesion disease according to RECIST (Response Evaluation Criteria in Solid Tumors) versions 1.1 (refer to the literature [Eisenhauer EA, Therasse P, Bogaerts J, Schwartz LH, Sargent D, Ford R, et al. New response evaluation criteria in solid tumors: revised RECIST guideline (Version 1.1). Eur J Cancer. 2009; 45:228 247]).

[0180] 5) Must be able to provide tumor tissue samples deemed suitable for biomarker analysis (in the case of hepatocellular carcinoma, registration is possible even if there is no tumor tissue sample before administration of the test drug).

[0181] General selection criteria

[0182] 6) 19 years of age or older

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

[0184]

[0185] 8) You must sign the consent form.

[0186] 9) For men and women of childbearing age, those who can adhere to appropriate contraception during treatment and for up to 3 months after the last treatment administration

[0187] Exclusion criteria

[0188] Subjects who met any of the following criteria were excluded from this clinical trial.

[0189] Tumor-related exclusion criteria

[0190] 1) Radically unresectable or metastatic disease

[0191] 2) Previous treatment for gastric adenocarcinoma, esophageal squamous cell carcinoma, or hepatocellular carcinoma. However, for hepatocellular carcinoma, even if prior treatment was given for a localized lesion, if more than 6 months have passed since the treatment and the treated area has progressed, or if a new lesion has developed outside of the previously treated area and is amenable to radical resection, and other inclusion / exclusion criteria are met, clinical trial registration is possible.

[0192] 3) Patients with a history of another cancer within the three years prior to the start of trial treatment. However, patients with cancer that has a minor impact on the patient's prognosis, such as intraepithelial carcinoma or papillary thyroid carcinoma, may be enrolled at the investigator's discretion.

[0193] 4) History of hepatic encephalopathy

[0194] 5) Clinically significant ascites defined as:

[0195] - Ascites or ascites found in the physical examination during screening

[0196] - Past ascites that required treatment and ongoing prevention or current ascites that require treatment

[0197] Exclusion criteria for clinical trial drugs

[0198] 6) History of active autoimmune disease requiring systemic treatment (i.e., disease-modifying agents, corticosteroids, or immunosuppressants) within the past two years. Replacement therapies (e.g., thyroxine or insulin, or physiologic corticosteroid replacement therapy for adrenal or pituitary insufficiency) are not considered systemic treatment and are permitted.

[0199] 7) Have been diagnosed with immunodeficiency or have been receiving chronic systemic steroid therapy (in doses exceeding the equivalent of 10 mg of prednisone daily) or any other form of immunosuppressive therapy within 7 days prior to the first dose of the clinical trial drug.

[0200] 8) History of or current interstitial pneumonia (non-infectious) requiring steroid treatment.

[0201] 9) Previous treatment with anti-PD-1, anti-PD-L1, anti-PD-L2, anti-CTLA-4 antibodies, or other antibodies or drugs specifically targeting T cell costimulation or checkpoint pathways.

[0202] 10) Known severe hypersensitivity or anaphylaxis to recombinant proteins, including monoclonal antibodies.

[0203] General exclusion criteria

[0204] 11) Have an active infection requiring systemic treatment

[0205] 12) There is a history or current evidence of a condition, treatment, or laboratory abnormality that, in the opinion of the treating investigator, could confound the results of the clinical trial, interfere with the participant's participation throughout the clinical trial, or indicate that participation in the clinical trial would not be in the participant's best interest.

[0206] 13) Women of childbearing potential who tested positive in a urine or blood pregnancy test within 7 days prior to the first administration of the test drug

[0207] 14) Are pregnant or breastfeeding, or plan to conceive or have a second child during the intended clinical trial period up to 90 days after the final administration of the clinical trial treatment.

[0208] 15) Symptomatic congestive heart failure (i.e., New York Heart Association class II or higher) or history or current evidence of clinically significant cardiac arrhythmia requiring treatment with antiarrhythmic agents other than beta-blockers or digoxin and / or conduction abnormalities (except atrial fibrillation and paroxysmal ventricular tachycardia) within 6 months prior to study treatment, active coronary artery disease, unstable angina, new-onset angina within 3 months of study entry, or myocardial infarction within 6 months of study entry.

[0209] 16) Known history of human immunodeficiency virus (HIV (HIV 1 / 2 antibodies))

[0210] 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, subjects with hepatitis B are permitted to participate in the clinical trial if HBV DNA is <500 IU / mL (or 2500 copies / mL) at screening. Subjects with detected HBsAg or HBV DNA should be managed according to treatment guidelines. Subjects receiving antiviral agents at screening must have been receiving treatment for at least 2 weeks prior to enrollment and must continue treatment for 6 months after starting the study drug. Patients with HCV antibody positivity can be enrolled only if they are HCV RNA negative by PCR (exception: subjects with hepatocellular carcinoma can be enrolled regardless of HCV RNA positivity).

[0211] 18) Have a history of allogeneic tissue / solid organ transplantation

[0212] 19) Received a live vaccine within 28 days prior to the first dose of the test treatment

[0213] Experimental treatment (prior IMC-001 treatment before surgery)

[0214] Methods of preparing and administering clinical trial drugs

[0215] IMC-001 was supplied as an injection in a sterile glass vial filled with 10 mL of 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 as an intravenous infusion over 60 minutes (±10 minutes). Subjects were carefully monitored for infusion reactions during IMC-001 administration. Premedication with antihistamines, antipyretics, and / or analgesics (e.g., acetaminophen) was permitted and could be administered at the investigator's discretion. If an infusion-related reaction occurred, subjects were managed according to the treatment guidelines. Vital signs (blood pressure, pulse rate, respiratory rate, and body temperature) were measured within 60 minutes before the infusion. If clinically indicated, vital signs were measured every 15 minutes (±10 minutes) during the infusion or 30 minutes (±10 minutes) after the infusion. If the subject experienced an infusion-related reaction during the first infusion, vital signs were measured during the infusion and 30 minutes (±10 minutes) after the infusion.

[0216] Initial dose and dosing schedule

[0217] The dose of IMC-001 in this clinical trial was 20 mg / kg, which was determined as the recommended dose for phase 2 clinical trials (RP2D) in an ongoing phase 1 clinical trial. It was administered as an intravenous infusion on day 1 (±4 days) of every 14-day cycle. Dose calculation was based on baseline body weight. If a subject's body weight was within 10% of the weight used in the previous dose calculation, the dose was not recalculated. All doses were rounded to the nearest milligram (mg) according to the clinical trial site's criteria.

[0218] Total administration period

[0219] A total of two cycles of preoperative neoadjuvant IMC-001 were administered unless patients were prematurely withdrawn or withdrew consent due to unacceptable toxicity or cancer progression, or continued treatment was deemed detrimental.

[0220] surgery:

[0221] Surgery was performed within 4 weeks after completion of the second cycle of neoadjuvant IMC-001 (days 11 to 42 from the day of IMC-001 administration [day 1] of the second cycle). However, the date of the visit at the end of the neoadjuvant treatment preceded the date of surgery. At this time, the subjects had a white blood cell count greater than 3,000 / μL, a platelet count greater than 75,000 / μL, no Grade 2 or higher toxicity due to the investigational drug treatment, and were clinically fit to undergo major surgery. Surgical procedures were performed according to institutional treatment guidelines for gastric, esophageal, and hepatocellular carcinoma. Open surgery was considered the standard approach. Laparoscopic and robot-assisted surgeries were permitted after discussion with the principal investigator and were performed only by a surgical team with extensive professional experience who considered laparoscopic and robot-assisted surgeries to be equivalent to open approaches in terms of morbidity, mortality, and oncologic outcomes. Laparoscopic surgery for clinical trials could be considered by the surgical team during the initial evaluation, after reviewing the extent and resectability of the tumor and evidence of distant metastases.

[0222] resection

[0223] The extent of surgical resection was determined based on tumor location, size, and progression. The resectability of the cancer was determined intraoperatively by the performing surgeon. The following factors could make resectability impossible:

[0224] - Remote metastasis

[0225] - Tumors that have invaded the vascular structures and / or other organs

[0226] - In cases where other radical resections are deemed difficult

[0227] Typically, if complete resection (R0) was not possible, the surgical team decided whether to perform non-curative surgery.

[0228] lymphadenectomy

[0229] The extent of lymphadenectomy was determined according to institutional treatment guidelines for gastric cancer, esophageal cancer, and hepatocellular carcinoma.

[0230] Postoperative care

[0231] Treatment of patients who underwent adjuvant chemotherapy after radical resection or non-curative resection or who could not undergo resection was performed according to the institutional guidelines.

[0232] Drugs and treatments prohibited for concurrent use

[0233] The following drugs and treatments were prohibited during clinical trials (except when used to treat adverse reactions related to the trial drug):

[0234] - During the period of administration of the preceding anticancer therapy administered in this clinical trial, anticancer therapy for the purpose of treating cancer (including anticancer chemotherapy, hormone therapy, immunotherapy, radiotherapy, biological anticancer therapy, or herbal medicine) excluding the clinical trial drug

[0235] - Other clinical trial treatments within 28 days prior to and during administration of the clinical trial drug used in this clinical trial

[0236] - Live attenuated vaccines (e.g., FluMist®) within 28 days before starting clinical trial drug administration, during clinical trial drug administration, and for 5 months after the last dose of clinical trial drug

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

[0238] - Systemic corticosteroids: The use of corticosteroids exceeding 10 mg / day of prednisone or equivalent was generally not permitted. Corticosteroids could temporarily exceed 10 mg / day when medically necessary, such as to control symptoms of adverse reactions such as infusion reactions or irAEs, or before computed tomography (CT) scans in subjects with a history of allergy to intravenous contrast media. If the corticosteroid dose could not be reduced to a prednisone dose of 10 mg / day or less within 12 weeks of starting steroids, the investigator promptly initiated a discussion about the subject's continuation of study treatment.

[0239] Concurrently Allowed Medications and Treatments

[0240] The following medications and treatments were permitted:

[0241] - Oral contraceptives

[0242] - Hormone replacement therapy

[0243] - Inactivated flu vaccine

[0244] - Megestrol acetate administered as an appetite stimulant

[0245] - Topical, ocular, intra-articular, nasal, and inhaled corticosteroids

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

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

[0248] The use of other medications necessary for subject management was at the investigator's discretion. If nausea, vomiting, or diarrhea occurred, effective symptomatic treatment was initiated. Antiemetics were administered to subjects according to local institutional guidelines or other guidelines (e.g., ASCO guidelines). Hematopoietic growth factors (G-CSF or GM-CSF) were used for the treatment of febrile neutropenia according to institutional guidelines or other guidelines (e.g., ASCO guidelines), but were used for primary prevention. Paracetamol (acetaminophen), meperidine, and chlorpheniramine or other antihistamines were used for the treatment of infusion reactions associated with the study drug according to institutional guidelines.

[0249] Assessment of severity of adverse reactions

[0250] The severity of adverse reactions was assessed using the Adverse Reaction Severity Grading Scale based on the NCI Common Terminology Criteria (CTCAE) v4.03 (http: / ctep.cancer.gov / protocolDevelopment / electronic_applications / ctc.htm#ctc_40). Table 6 was used to assess the severity of adverse reactions not listed in the NCI CTCAE. Heart failure was classified according to the New York Heart Association (NYHA) classification criteria.

[0251]

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

[0253] Safety

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

[0255] Clinical tumor response

[0256] No patient experienced disease progression during neoadjuvant therapy. Among 17 patients with measurable lesions by RECIST 1.1, 3 (17.6%) achieved partial response (PR) and 14 (82.4%) achieved stable disease (SD) (Figure 2).

[0257] Among 30 patients for whom metabolic response could be assessed by EORTC criteria, 7 (23.3%) achieved metabolic partial remission (mPR), 20 (66.7%) achieved metabolic stable disease (mSD), and 3 (10.0%) achieved metabolic progressive disease (mPD) ( Figure 3 ). Two of the three patients with metabolic progressive disease demonstrated increased tumor-infiltrating lymphocytes and tumor regression on surgical pathology.

[0258] Surgical results

[0259] All patients underwent radical resection (R0 resection) of stage I (n = 36, 75.0%) / II (n = 6, 12.5%) / III (n = 6, 12.5%). There was no operative mortality or treatment-related perioperative morbidity.

[0260] Pathological reaction

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

[0262] relapse

[0263] The median follow-up period was 31.4 months (range: 1.7 to 41.2 months), with only 4 patients (8.3%) relapsed and the median progression-free survival (PFS) or relapse-free survival (RFS) not reached.

[0264] conclusion

[0265] IMC-001 neoadjuvant therapy was well tolerated and showed promising antitumor activity in patients with microsatellite stable, resectable GC, EC, and HCC.

Claims

1. A pharmaceutical composition for treating tumors or inhibiting tumor proliferation in patients with upper gastrointestinal cancer, A therapeutically effective amount of an anti-PD-L1 antibody that specifically binds to PD-L1 and comprises HCDR1, HCDR2 and HCDR3 contained in a heavy chain variable region (HCVR) of SEQ ID NO: 1 and LCDR1, LCDR2 and LCDR3 contained in a light chain variable region (LCVR) of SEQ ID NO: 2, The above antibody is administered to the patient as neoadjuvant therapy prior to surgical resection of the tumor in the patient. Pharmaceutical composition.

2. In paragraph 1, The above upper digestive tract cancer is stomach cancer, esophageal cancer, or liver cancer. Pharmaceutical composition.

3. In paragraph 1, The above anti-PD-L1 antibody comprises 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. Pharmaceutical composition.

4. In paragraph 3, The light chain variable region comprises the amino acid sequence of SEQ ID NO: 2 or an amino acid sequence that is at least 95% identical to SEQ ID NO:

2. Pharmaceutical composition.

5. In paragraph 4, The amino acids at positions 3 and 5 of the light chain variable region are identical to the amino acids at positions 3 and 5 of SEQ ID NO:

2. Pharmaceutical composition.

6. In paragraph 4, The above heavy chain variable region comprises an amino acid sequence that is at least 95% identical to SEQ ID NO:

1. Pharmaceutical composition.

7. In paragraph 1, The above anti-PD-L1 antibody comprises a heavy chain having an amino acid sequence of SEQ ID NO: 3 and a light chain having an amino acid sequence of SEQ ID NO:

4. Pharmaceutical composition.

8. In paragraph 1, The above upper gastrointestinal cancer is resectable. Pharmaceutical composition.

9. In paragraph 1, The above upper gastrointestinal cancer is recurrent Pharmaceutical composition.

10. In paragraph 1, The above upper gastrointestinal cancer is metastatic. Pharmaceutical composition.

11. In paragraph 1, The purpose of the surgery for the above upper gastrointestinal cancer is radical treatment. Pharmaceutical composition.

12. In paragraph 2, The above gastric cancer is gastric submucosal tumor or gastric adenocarcinoma (GC). Pharmaceutical composition.

13. In paragraph 2, The above esophageal cancer is esophageal squamous cell carcinoma (ESCC). Pharmaceutical composition.

14. In paragraph 2, The above liver cancer is hepatocellular carcinoma (HCC). Pharmaceutical composition.

15. In paragraph 1, As the above neoadjuvant therapy, anti-PD-L1 antibody is administered in one or more doses before surgical resection of the tumor. Pharmaceutical composition.

16. In paragraph 15, Each of the above doses is administered at intervals of 10 to 18 days. Pharmaceutical composition.

17. In paragraph 16, As the above neoadjuvant therapy, anti-PD-L1 antibody is administered 2 to 6 times at 10 to 18-day intervals before surgical resection of the tumor. Pharmaceutical composition.

18. In paragraph 1, As the above neoadjuvant therapy, anti-PD-L1 antibody is administered at a dose of 10 mg / kg to 30 mg / kg. Pharmaceutical composition.

19. In paragraph 18, As the above neoadjuvant therapy, anti-PD-L1 antibody is administered at a dose of 20 mg / kg. Pharmaceutical composition.

20. In paragraph 1, Surgical resection of the above tumor is performed between 11 and 84 days after the last administration of anti-PD-L1 antibody. Pharmaceutical composition.

21. As a method for treating tumors or inhibiting tumor growth, (a) A step for screening patients with upper gastrointestinal cancer; (b) administering a PD-L1 inhibitor to the patient in a therapeutically effective amount; and (c) a step of surgically resecting the upper gastrointestinal cancer tumor after step (b); The above PD-L1 inhibitor is an 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. method.

22. A kit comprising a PD-L1 inhibitor together with instructions for use of the PD-L1 inhibitor as neoadjuvant therapy for treating tumors or inhibiting tumor growth in patients with upper gastrointestinal cancer, The above instructions include instructions for administering the PD-L1 inhibitor intravenously at a dose of 20 mg / kg for 2 to 6 doses at intervals of 10 to 18 days prior to surgical resection of the tumor. Kit.

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