CN111973739A

CN111973739A - Use of anti-PD-L1 monoclonal antibody for treating cancer

Info

Publication number: CN111973739A
Application number: CN202010407559.1A
Authority: CN
Inventors: 杨朝强; 苏楠; 张喜全; 王训强
Original assignee: Chia Tai Tianqing Pharmaceutical Group Co Ltd; Nanjing Shunxin Pharmaceutical Co Ltd
Current assignee: Chia Tai Tianqing Pharmaceutical Group Co Ltd; Nanjing Shunxin Pharmaceutical Co Ltd
Priority date: 2019-05-23
Filing date: 2020-05-14
Publication date: 2020-11-24
Anticipated expiration: 2040-05-14
Also published as: CN111973739B

Abstract

The invention provides the use of an anti-PD-L1 monoclonal antibody for the treatment of cancer, in particular for the treatment of mediastinal large B-cell lymphoma, comprising administering to said subject a therapeutically effective amount of an inhibitor of the interaction between the PD-1 receptor and its ligand PD-L1, wherein the inhibitor is an anti-PD-L1 monoclonal antibody, clinical trial results showing significant control of disease progression.

Description

Use of anti-PD-L1 monoclonal antibody for treating cancer

Technical Field

The present invention provides a method of treating cancer in a subject comprising administering to the subject a therapeutically effective amount of an inhibitor of the interaction between the PD-1 receptor and its ligand PD-L1.

Background

The natural immune system containing T lymphocytes has strong anticancer ability and has wide applicationPan-competence and fine specificity, responding to various tumor antigens. Emerging cancer immunotherapy enhances anti-tumor immune responses by adoptive transfer of activated effector cells, immunization against relevant antigens, or provision of non-specific immunostimulants. Over the past decade, researchers have struggled to develop specific immune checkpoint inhibitors and expect to provide new immunotherapeutic protocols for the treatment of cancer, including the development of antibodies (antibodies) Ipilimumab (Ipilimumab) that bind to and inhibit CTLA-4

For treating patients with advanced melanoma (Hodi et al (2010) N Engl J Med 363:711-23), and for developing antibodies such as Nivolumab

And Pabolilizumab (Pembrolizumab)

They specifically bind to programmed death receptor-1 (PD-1) and block the inhibitory PD-1/PD-1 ligand pathway (Topalian et al (2012a) N Engl J Med 366: 2443-54). Among them, PD-1(programmed death-1) is a key immune checkpoint receptor expressed by activated T lymphocytes and B lymphocytes and mediates immunosuppression, and its ligands include at least PD-L1 and PD-L2.

PD-L1(Programmed death-ligand 1), also known as CD274 and B7-H1, is a 40kDa type 1 transmembrane protein encoded by the CD274 gene and is a ligand for PD-1. Both PD-L1 and PD-1 belong to the immunoglobulin superfamily and both consist of two extracellular Ig domains, an N-terminal V domain and a C-terminal constant domain. The binding interface of PD-L1 to programmed death receptor 1(PD-1) and B7-1(CD80) is on an IgV-like domain (Lin et al (2008) PNAS 105: 3011-3016). While PD-L1 contains a conserved short intracellular tail region (about 30 amino acids), PD-1 contains two cytoplasmic tyrosine-based signaling motifs, namely an immunoreceptor tyrosine-based inhibitory motif (ITIM) and an immunoreceptor tyrosine-based switching motif (ITSM). Following T cell stimulation, PD-1 recruits the tyrosine phosphatase SHP-2 to the ITSM motif in its cytoplasmic tail, resulting in dephosphorylation of effector molecules involved in the CD 3T cell signaling cascade, such as CD3 ζ, PKC θ and ZAP70 (Freeman et al (2000) J Exp Med 192: 1027-34; Latchman et al (2001) Nat Immunol 2: 261-8; Carter et al (2002) Eur J Immunol32: 634-43). PD-L1 is widely distributed not only on leukocytes and nonhematopoietic cells in lymphoid and non-lymphoid tissues, but also in various cancer cells, is highly expressed on the surface of various tumor cells, and the degree of malignancy and poor prognosis of tumors are closely related to the expression level of PD-L1. There are clinical data indicating that high tumor expression of PD-L1 is associated with increased tumor invasiveness and poor prognosis. The formation of the PD-1/PD-L1 complex transmits inhibitory signals and negatively regulates T cell immune responses; it inhibits TCR-mediated T cell activation, cytokine production and T cell proliferation (Fife et al (2011) Nature Immunology 10: 1185-1193); induction of depletion or anergy in cognate antigen-specific T cells (Hofmeyer et al (2011) Journal of Biomedicine and Biotechnology 2011: 1-9); promote the differentiation of Th1 cells into Foxp3+ regulatory T cells (Armanath et al (2011) Science TransMed 3: 1-13; Francisco et al (2009) J. exp. Med.206: 3015-; and inducing apoptosis of effector T cells. Disruption of the PD-L1 gene resulted in an upregulated T cell response and the generation of autoreactive T cells (Latchman et al (2004) PNAS 101: 10691-10696). Antibody blockade of PD-1 or PD-L1 resulted in increased anti-tumor immunity (Iwai et al (2002) PNAS99: 12293-12297).

The anti-PD-L1 antibody can block the interaction of PD-L1 with PD-1 and CD80, so that the related negative regulation signals can not be started and conducted, thereby preventing the activity of effector T cells in a tumor microenvironment from being inhibited, and enabling the T cells to play the functions of killing and inhibiting the tumor cells. The anti-PD-L1 antibody can directly act on tumor tissues, so that the antibody has high specificity and safety. The major anti-PD-L1 monoclonal antibody drug products currently include Atezolizumab, Roche, Durvalumab, Asricon, and Avelumab, Merck & Hurride, among others. Patent WO2016022630 also discloses an anti-PD-L1 antibody, which has high affinity for PD-L1, can significantly inhibit the interaction between PD-L1 and PD-1 on the cell surface, and significantly promote the secretion of IL-2 and IFN-gamma by T cells.

Malignant lymphoma is an immune cell tumor of lymph nodes and lymph tissue outside the node, derived from malignant changes of lymphocytes or histiocytes. Malignant lymphomas are classified into Hodgkin's Disease (HD) and non-hodgkin's lymphoma (NHL). In China, the incidence rate of malignant lymphoma is 11-13 malignant tumors, and the incidence rate is increased in recent years, which at least exceeds 25000 cases per year. The incidence in western countries such as europe, america and australia can be as high as 11/10-18/10 ten thousand, slightly higher than the sum of all types of leukemia. At least 3 million new cases are found each year in the united states. At present, the treatment of malignant lymphoma is being changed from the conventional treatment mainly involving nonspecific cytotoxic drug chemotherapy and radiotherapy to multidisciplinary comprehensive treatment involving various modes such as biological targeted therapy. NHL is further classified into aggressive lymphoma and indolent lymphoma. Primary mediastinal large B-cell lymphoma (PMBCL) is an aggressive histological subtype of NHL, accounting for approximately 5% of aggressive lymphoma, 2% of all lymphomas. Compared with diffuse large B-cell lymphoma (DLBCL), PMBCL has a unique molecular marker, and shares with tuberous-sclerosis type non-hodgkin's lymphoma. PMBCL is mainly seen in women, most of the tumors are better to be developed at the anterior mediastinum, the growth speed is higher, the cases that the diameter of the tumor is larger than 10cm are more common, and the PMBCL is often accompanied with the symptoms of infiltration of the respiratory system and partial thoracic cavities (lung, chest wall, pleura and pericardium). In 50% of patients, symptoms and signs of superior vena cava syndrome can occur, such as facial edema, jugular vein anger, and upper limb edema and/or deep vein thrombosis are sometimes accompanied. Approximately 20% of patients are associated with systemic symptoms, primarily fever and weight loss, and 70% of patients can develop elevated Lactate Dehydrogenase (LDH). Most patients are in stage I-II at the time of initial treatment, and only 20% of patients are in stage III-IV. Distant metastasis spreading outside the thorax is not common in naive patients, but in recurrent patients, the kidneys, adrenal glands, liver, ovaries and central nervous system are affected, but bone marrow infiltration is rare. Relapsed/refractory primary mediastinal large B-cell lymphoma (rrPMBCL) treatment is generally referred to relapsed/refractory diffuse large B-cell lymphoma, and is recommended for patients who relapse after autologous stem cell transplantation therapy or who are not amenable to large dose treatment to participate in clinical trials or receive second-line chemotherapy regimens, palliative radiation therapy, or supportive care. In a study of 106patients with PMBCL (Lazzarino M, Orlandi E, Pauli M, et al.treatment out of mice and systemic factors for primary medical (thymic) B-cell lymphoma: a multicenter study of 106patients. J Clin Oncol.1997; 15(4): 1646) 1653.), doxorubicin-containing treatment resulted in poor results of rrPMBCL, the effective rate of 35 refractory PMBCL was 0%, and only 4 of 18 patients with relapsed PMBCL was effective (22%); another study for the treatment of rrPMBCL/rrDLBCL (Kuruvilla J, Pintilie M, Tsang R, Nagy T, Keting A, Crump M. Salvage chemotherapy and autogous stem cell transplantation area information for replayed or recurrent primary therapy large B-cell Lymphoma compounded with two drug large B-cell Lymphoma. Leuk Lymphoma.), rrPMBCL rescue chemotherapy had a total efficacy of 25%.

Disclosure of Invention

Summary of the invention

The present invention provides a method of treating cancer in a subject comprising administering to the subject a therapeutically effective amount of an inhibitor of the interaction between the PD-1 receptor and its ligand PD-L1, wherein the inhibitor is a PD-L1 antibody.

The invention also provides the use of an anti-PD-L1 antibody for the treatment of cancer, comprising administering to the subject a therapeutically effective amount of an inhibitor of the interaction between the PD-1 receptor and its ligand PD-L1, wherein the inhibitor is an anti-PD-L1 antibody.

The invention also provides the use of an anti-PD-L1 antibody in the manufacture of a medicament for the treatment of cancer.

In some embodiments, the anti-PD-L1 antibody comprises the amino acid sequence: a heavy chain CDR1 region having at least 80% homology to the amino acid sequence set forth in SEQ ID NO.1 or SEQ ID NO. 4; a heavy chain CDR2 region having at least 80% homology to the amino acid sequence set forth in SEQ ID NO.2 or SEQ ID NO. 5; a heavy chain CDR3 region having at least 80% homology to the amino acid sequence set forth in SEQ ID NO. 3 or SEQ ID NO. 6; a light chain CDR1 region having at least 80% homology to the amino acid sequence set forth in SEQ ID NO. 7 or SEQ ID NO. 10; a light chain CDR2 region having at least 80% homology to the amino acid sequence set forth in SEQ ID NO. 8 or SEQ ID NO. 11; a light chain CDR3 region having at least 80% homology to the amino acid sequence set forth in SEQ ID NO. 9 or SEQ ID NO. 12.

In some embodiments, the anti-PD-L1 antibody is administered at a dose of 1mg/kg, 2mg/kg, 3mg/kg, 5mg/kg, 6mg/kg, 9mg/kg, 10mg/kg, 15mg/kg, 20mg/kg, 30mg/kg body weight for continuous dosing.

In some embodiments, the anti-PD-L1 antibody is administered in one or more unitary doses that are effective to treat the cancer. In some embodiments, wherein the unitized dose is in the range of about 20mg to about 2000mg of anti-PD-L1 antibody. In some embodiments, wherein the unitized dose is selected from about 300mg, about 600mg, about 900mg, about 1000mg, about 1200mg, about 1500mg, about 1800mg, about 2100mg, or about 2400mg of the anti-PD-L1 antibody. In some embodiments, the unitized dose is selected from about 1200mg of anti-PD-L1 antibody.

In some embodiments, the anti-PD-L1 antibody is administered about every week (q1w), about every 2 weeks (q1w), about every 3 weeks (q1w), or about every 4 weeks (q1 w). In some embodiments, the patient is administered a uniform dose of anti-PD-L1 antibody about every 3 weeks. In some embodiments, the anti-PD-L1 antibody is administered at a dose of 1200mg per patient, approximately once every 3 weeks, for a sustained period.

In some embodiments, the anti-PD-L1 antibody is administered as an intravenous infusion. In some embodiments, the anti-PD-L1 antibody is administered as an intravenous infusion over about 1-2 hours, preferably about 1 hour.

In some embodiments, the anti-PD-L1 antibody is a naked antibody, an intact antibody, or an antibody fragment comprising an antigen binding region.

In some embodiments, the method results in an objective response, preferably a complete response or a partial response.

In some embodiments, the subject has previously received surgery, chemotherapy, and/or radiation therapy. In some embodiments, the subject has reoccured disease progression after achieving complete remission post-surgery. In some embodiments, the subject has failed to provide complete or partial remission following surgery. In some embodiments, the subject regains disease progression after achieving complete remission following chemotherapy. In some embodiments, the subject has failed to complete remission or failed to partial remission following chemotherapy. In some embodiments, the subject regains disease progression after achieving complete remission following radiation therapy. In some embodiments, the subject has failed to complete remission or failed to partial remission following radiation therapy. In some embodiments, the subject has previously received surgery and chemotherapy. In some embodiments, the subject has reoccured disease progression after achieving complete remission following surgery and chemotherapy. In some embodiments, the subject has failed to complete remission or failed to partial remission following surgery or chemotherapy. In some aspects, the subject has previously undergone surgery and radiation therapy. In some embodiments, the subject has reoccured disease progression after achieving complete remission following surgery and radiation therapy. In some embodiments, the subject has failed to alleviate completely or partially after surgery and radiation therapy. In some embodiments, the subject has previously received chemotherapy and radiation therapy. In some embodiments, the subject is treated with chemotherapy and radiation to achieve complete remission before disease progression occurs. In some embodiments, the subject has failed to complete remission or failed to partial remission following chemotherapy and radiation therapy. In some embodiments, the subject has previously received surgery, chemotherapy, and radiation therapy. In some embodiments, the subject has re-developed disease progression after achieving complete remission following surgery, chemotherapy, and radiation therapy. In some embodiments, the subject has failed to complete remission or partial remission following surgery, chemotherapy, and radiation therapy.

In some embodiments, the subject has undergone autologous stem cell transplantation following surgery, chemotherapy, and/or radiation therapy. In some embodiments, the subject regains disease progression after obtaining complete remission following surgery, chemotherapy, and/or autologous stem cell transplantation. In some embodiments, the subject has failed to complete remission or failed to partial remission following surgery, chemotherapy, and/or autologous stem cell transplantation. In some embodiments, the subject has undergone an autologous stem cell transplant following surgery. In some embodiments, the subject has reoccured disease progression after surgery and after complete remission from autologous stem cell transplantation. In some embodiments, the subject has failed to achieve complete remission or partial remission following surgery or autologous stem cell transplantation. In some embodiments, the subject has received an autologous stem cell transplant following receiving chemotherapy. In some embodiments, the subject has re-developed disease progression after complete remission following chemotherapy and autologous stem cell transplantation. In some embodiments, the subject has failed to complete remission or partial remission following chemotherapy and autologous stem cell transplantation. In some embodiments, the subject has received autologous stem cell transplantation after receiving radiation therapy. In some embodiments, the subject has re-developed disease progression after complete remission following radiation therapy and autologous stem cell transplantation. In some embodiments, the subject has failed to complete remission or failed to partial remission following radiation therapy and autologous stem cell transplantation. In some embodiments, the subject has undergone autologous stem cell transplantation after undergoing surgery and chemotherapy. In some embodiments, the subject has reoccured disease progression after obtaining complete remission following surgery, chemotherapy, and autologous stem cell transplantation. In some embodiments, the subject has failed to complete remission or partial remission following surgery, chemotherapy, and autologous stem cell transplantation. In some embodiments, the subject has undergone autologous stem cell transplantation after undergoing surgery and radiation therapy. In some embodiments, the subject has re-developed disease progression after obtaining complete remission following surgery, radiation therapy, and autologous stem cell transplantation. In some embodiments, the subject has failed to complete remission or partial remission following surgery, radiation therapy, and autologous stem cell transplantation. In some embodiments, the subject has received autologous stem cell transplantation after receiving chemotherapy and radiation therapy. In some embodiments, the subject has re-developed disease progression after obtaining complete remission following chemotherapy, radiation therapy, and autologous stem cell transplantation. In some embodiments, the subject has failed to complete remission or failed to partial remission following chemotherapy, radiation therapy, and autologous stem cell transplantation.

In some embodiments, the subject has undergone an autologous stem cell transplant following surgery. In some embodiments, the subject has re-developed disease progression after complete remission following autologous stem cell transplantation. In some embodiments, the subject has failed to complete remission or failed to partial remission following transplantation of autologous stem cells.

In some embodiments, the cancer is non-Hodgkin's lymphoma. In some embodiments, the cancer is mediastinal large B-cell lymphoma. In some embodiments, the mediastinal large B-cell lymphoma has progressed following chemotherapy and/or radiation therapy. In some embodiments, the cancer is relapsed or refractory mediastinal large B-cell lymphoma. In some embodiments, the mediastinal large B-cell lymphoma is relapsed. In some embodiments, the mediastinal large B-cell lymphoma is refractory. In some embodiments, the mediastinal large B-cell lymphoma is metastatic. In some embodiments, the cancer treatment is second line treatment of relapsed or refractory mediastinal large B-cell lymphoma. In some embodiments, the cancer treatment is a second line treatment of metastatic mediastinal large B-cell lymphoma. In some embodiments, the relapsed or refractory mediastinal large B-cell lymphoma does not progress during completion of first-line therapy and/or after radiation therapy. In some embodiments, the metastatic mediastinal large B-cell lymphoma does not progress during completion of first-line therapy and/or after radiation therapy. In some embodiments, the relapsed or refractory mediastinal large B-cell lymphoma has progressed following chemotherapy and/or radiation therapy. In some embodiments, the metastatic mediastinal large B-cell lymphoma has progressed following chemotherapy and/or radiation therapy.

The invention provides an article of manufacture comprising a container containing a fixed dose of an anti-PD-L1 antibody. The invention also provides for the use of an anti-PD-L1 antibody in the manufacture of a product for the treatment of cancer comprising a container comprising a fixed dose of an anti-PD-L1 antibody. In some embodiments, the container is a vial. The fixed dose is selected from about 300mg, about 600mg, about 900mg, about 1000mg, about 1200mg, about 1500mg, about 1800mg, about 2100mg, and about 2400mg of the anti-PD-L1 antibody. In some embodiments, the article of manufacture further comprises a package insert or package insert instructing the user to administer the fixed dose to a cancer patient. In some embodiments, the article of manufacture comprises 1 or more than 1 vial containing about 300mg or 600mg of anti-PD-L1 antibody. In some embodiments, the article of manufacture comprises 1 vial containing about 300mg of the anti-PD-L1 antibody. In some embodiments, the article of manufacture comprises 1 vial containing about 600mg of anti-PD-L1 antibody.

Detailed description and preferred embodiments of the invention

The present invention provides a method for treating a subject having a cancer or tumor comprising administering to the subject a therapeutically effective amount of an inhibitor of the interaction between the PD-1 receptor and its ligand PD-L1.

The present invention also provides a method for treating a subject having a cancer or tumor, the method comprising administering to the subject a therapeutically effective amount of: an antibody or antigen-binding portion thereof that binds programmed death ligand 1(PD-L1) and/or inhibits PD-L1 activity.

The present invention also provides a monotherapy for treating a subject having a cancer or tumor, said method comprising administering to said subject separately a therapeutically effective amount of: an antibody or antigen-binding portion thereof that binds programmed death ligand 1(PD-L1) and/or inhibits PD-L1 activity.

The invention also provides a method of treating a subject having a cancer or tumor that is mediastinal large B-cell lymphoma, the method comprising: (i) measuring the level of PD-L1 in a sample from the subject, wherein the subject is PD-L1 positive, and (ii) administering to the subject a therapeutically effective amount of an anti-PD-L1 antibody or antigen-binding portion thereof.

The present invention provides a method for treating a subject having a cancer or tumor. In other embodiments, the subject is a patient that has been histopathologically diagnosed as mediastinal large B-cell lymphoma (MBCL). In other embodiments, the subject is a relapsed or refractory patient with primary mediastinal large B-cell lymphoma (PMBCL). In other embodiments, the subject is a patient with relapsed or refractory primary mediastinal large B-cell lymphoma. In other embodiments, the subject is a patient with relapsed primary mediastinal large B-cell lymphoma, where relapsed refers to recurrence of disease progression after treatment to complete remission. In other embodiments, the subject is a patient with primary mediastinal large B-cell lymphoma that is refractory to obtaining complete or partial remission after treatment. In other embodiments, the subject is a patient with metastatic mediastinal large B-cell lymphoma. In some embodiments, the metastasis is lymph node metastasis. In other specific embodiments, the metastasis is pleural and/or local chest wall metastasis.

In some embodiments, the subject is a relapsed and refractory patient with primary mediastinal large B-cell lymphoma (PMBCL). In some embodiments, the subject is a patient with metastatic relapsed and refractory mediastinal large B-cell lymphoma. In some embodiments, the subject is a patient with metastatic relapsed or refractory mediastinal large B-cell lymphoma.

The invention also provides a method for identifying a subject having mediastinal large B-cell lymphoma, the subject being suitable for anti-PD-L1 antibody therapy, the method comprising measuring the level of PD-L1 in a sample of the subject, and wherein the subject is administered a therapeutically effective amount of an anti-PD-L1 antibody, or an antigen-binding portion thereof.

The invention also provides a kit for treating a subject having B-cell lymphoma from mediastinum, the kit comprising: (a) an anti-PD-L1 antibody or antigen-binding portion thereof; (b) instructions for treating the tumor.

The present invention provides methods of treating cancer using one or more immune checkpoint inhibitors, such as an anti-PD-L1 antibody or antigen-binding portion thereof or an anti-PD-L1 antibody or antigen-binding portion thereof. In one embodiment, the cancer is a primary cancer. In one embodiment, the cancer is metastatic or recurrent cancer. In one embodiment, the cancer is a relapsed or refractory cancer. In one embodiment, the cancer is primary mediastinal large B-cell lymphoma. In one embodiment, the cancer is relapsed or refractory primary mediastinal large B-cell lymphoma.

The invention also provides the use of an anti-PD-L1 antibody in the first-line treatment of primary mediastinal large B-cell lymphoma. The invention also provides the use of an anti-PD-L1 antibody in the second-line treatment of primary mediastinal large B-cell lymphoma. The invention also provides an application of the anti-PD-L1 antibody in the three-line treatment of primary mediastinal large B cell lymphoma. In some embodiments, the anti-PD-L1 antibody is used for second-line treatment of relapsed or refractory primary mediastinal large B-cell lymphoma. In some embodiments, the anti-PD-L1 antibody is used for three-line therapy of relapsed or refractory primary mediastinal large B-cell lymphoma. In some embodiments, the anti-PD-L1 antibody is used alone for second-or third-line therapy of relapsed or refractory primary mediastinal large B-cell lymphoma. In some embodiments, the anti-PD-L1 antibody is used for subsequent treatment of relapsed or refractory primary mediastinal large B-cell lymphoma. In some embodiments, the anti-PD-L1 antibody is used alone for second-or third-line therapy in patients with relapsed or refractory primary mediastinal large B-cell lymphoma.

Treatment of primary mediastinal large B-cell lymphoma may be referred to NCCN guidelines 5 th edition at the end of 2017. Optimal first line therapy is more controversial than other subtypes of NHL, however, these treatment regimens include: dose-adjusted DA-EPOCH-R ([ etoposide, prednisone, vincristine, cyclophosphamide, doxorubicin ] + rituximab) x 6 cycles; RCHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, prednisone) x 6 cycle + radiotherapy; RCHOP 4 × followed by ICE (ifosfamide, carboplatin, etoposide) × 3 cycles + -radiation therapy, etc.

In certain embodiments, the subject is a human patient. In certain embodiments, the subject has received another cancer treatment (e.g., chemotherapy), but is resistant or refractory to such another cancer treatment.

In some embodiments, the subject has previously received EPOCH-R or RCHOP therapy. In some embodiments, the subject regains disease progression after complete remission following treatment with EPOCH-R or RCHOP. In some embodiments, the subject has failed to achieve complete or partial remission following treatment with EPOCH-R or RCHOP.

In certain embodiments, the invention provides a method for treating a subject having a tumor, the method comprising administering to the subject a therapeutically effective amount of an immune checkpoint inhibitor, e.g., an anti-PD-L1 antibody. In certain embodiments, the invention relates to a method of treating a subject having a tumor that is a relapsed or refractory primary mediastinal large B-cell lymphoma, the method comprising: (i) measuring the level of PD-L1 in a sample of the subject, wherein the subject expresses PD-L1, and (ii) administering to the subject a therapeutically effective amount of an immune checkpoint inhibitor, e.g., an anti-PD-L1 antibody or an antigen-binding portion thereof. In certain embodiments, the anti-PD-1 antibody is 13C5, 5G11, ch13C5-hIgG1, ch13C5-hIgG4, ch5G11-hIgG1, ch5G11-hIgG4, hu13C5-hIgG1, hu13C5-hIgG4, hu5G11-hIgG1, or hu5G11-hIgG4 monoclonal antibody. In other embodiments, the anti-PD-1 antibody competes for binding with 5G11 mab.

In some embodiments, the cancer or tumor expresses PD-L1. The PD-L1 status of a tumor in a subject can be measured prior to administration of any composition or using any method disclosed herein. In one embodiment, the tumor has a PD-L1 expression level of at least about 0.5%, at least about 0.6%, at least about 0.7%, at least about 0.8%, at least about 0.9%, 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 6%, at least about 7%, at least about 8%, at least about 9%, at least about 10%, at least about 11%, at least about 12%, at least about 13%, at least about 14%, at least about 15%, at least about 20%, or greater than at least about 20%. In another embodiment, the PD-L1 status of the tumor is at least about 1%. In other embodiments, the PD-L1 status of the subject is at least about 5%. In a particular embodiment, the PD-L1 status of the tumor is at least about 10%. Measurement of PD-L1 status can be performed using methods such as antibodies, in situ mRNA hybridization, automated IHC, and the like.

The present invention provides a method of treating a subject for cancer or a tumor, the method comprising administering to the subject a therapeutically effective amount of an anti-PD-L1 antibody, or antigen-binding portion thereof, wherein a tumor sample obtained from the patient has been determined to have a detectable expression level of PD-L1 in cells comprising about 1% or more (e.g., about 1%, about 2%, about 3%, or about 4% or more). In some embodiments, the tumor sample obtained from the patient has been determined to have a detectable expression level of PD-L1 in cells comprising about 1% to about 65% or more (e.g., about 1% to about 5%, about 5% to about 10%, about 10% to about 20%, about 20% to about 30%, about 30% to about 40%, about 40% to about 50%, or about 50% to about 65%). The invention also provides a method of determining whether a patient suffering from primary mediastinal B-cell lymphoma is likely to respond to treatment with a therapeutic agent comprising an anti-PD-L1 antibody, the method comprising determining the expression level of PD-L1 in tumor cells in a tumor sample obtained from the patient, wherein a detectable expression level of PD-L1 in cells comprising about 1% or more of the tumor sample indicates that the patient is likely to respond to treatment with a therapeutic agent comprising an anti-PD-L1 antibody. The invention also provides a method for predicting responsiveness of a patient suffering from primary mediastinal B-cell lymphoma to treatment with a therapeutic agent comprising an anti-PD-L1 antibody, the method comprising determining the expression level of PD-L1 in cells in a tumor sample obtained from the patient, wherein a detectable expression level of PD-L1 in tumor cells comprising about 1% or more of the tumor sample indicates that the patient is likely to respond to treatment with the therapeutic agent comprising the anti-PD-L1 antibody. The invention also provides a method for selecting a therapy for a patient suffering from primary mediastinal B-cell lymphoma, the method comprising determining the expression level of PD-L1 in tumor cells in a tumor sample obtained from the patient, and selecting a therapy for the patient comprising a therapeutic agent comprising an anti-PD-L1 antibody based on the detectable expression level of PD-L1 in tumor cells comprising about 1% or more of the tumor sample. In some embodiments, the tumor sample obtained from the patient has been determined to have a detectable expression level of PD-L1 in tumor cells comprising about 5% or more of the tumor sample. In some embodiments, the tumor sample obtained from the patient has been determined to have a detectable expression level of PD-L1 in tumor cells comprising at least about 10% of the tumor sample. The invention also provides a method for determining whether a patient suffering from primary mediastinal large B-cell lymphoma is likely to respond to treatment with a therapeutic agent comprising an anti-PD-L1 antibody or antigen-binding portion thereof, the method comprising determining a subtype of the tumor from a tumor sample obtained from the patient, wherein a relapsed and/or refractory primary mediastinal large B-cell lymphoma indicates that the patient is likely to respond to treatment with a therapeutic agent comprising an anti-PD-L1 antibody. The present invention provides a method for predicting responsiveness of a patient suffering from primary mediastinal large B-cell lymphoma to treatment with a therapeutic agent comprising an anti-PD-L1 antibody, the method comprising determining a subtype of the tumor from a tumor sample obtained from the patient, wherein a relapsed and/or refractory primary mediastinal large B-cell lymphoma indicates that the patient is likely to respond to treatment with the therapeutic agent comprising an anti-PD-L1 antibody. The invention provides a method for selecting a therapy for a patient suffering from primary mediastinal large B-cell lymphoma, the method comprising determining a subtype of the tumor from a tumor sample obtained from the patient, and selecting a therapy for the patient comprising a therapeutic agent comprising an anti-PD-L1 antibody based on the primary mediastinal large B-cell lymphoma determined to be relapsed and/or refractory to the tumor. In some embodiments, the method further comprises administering to the patient a therapeutically effective amount of a therapeutic agent that is an anti-PD-L1 antibody based on the expression level of PD-L1 in tumor cells in the tumor sample.

In certain embodiments, the expression level of at least one of CD15, CD30, PAX5, CD20, EBV-EBER, CCL17, and/or CCL22 in a tumor sample or blood tissue sample obtained from the patient has been determined to be altered, e.g., elevated, relative to a reference level of the at least one gene; and/or, the expression level of at least one of CD15, CD45, CD20, CD99, and/or CD3 in a tumor sample obtained from the patient has been determined to be altered, e.g., decreased, relative to a reference level of the at least one gene. In other embodiments, the expression level of miR135a in the tumor sample obtained from the patient has been determined to be altered relative to a reference level of the microrna (micro rna), in some specific embodiments, the alteration is a decrease in the expression level. In some embodiments, the reference level is a non-tumor sample or a blood tissue sample of the non-diseased subject as a reference. In some embodiments, the reference level is a non-tumor sample of the patient as a reference.

In certain embodiments, the level of genetic mutation, level of genetic modification, level of transcription, and/or level of expression of at least one of PD-L1, TMB, MSI, 9p24.1 in a tumor sample or blood tissue sample obtained from the patient has been determined to be altered relative to a reference level of the at least one gene. In some embodiments, the reference level is a non-tumor sample or a blood tissue sample of the non-diseased subject as a reference. In some embodiments, the reference level is a non-tumor sample of the patient as a reference.

In certain embodiments, the objective response rate of a subject administered a therapeutically effective amount of an anti-PD-L1 antibody is between about 10% to about 40% (e.g., about 10% to about 20%, about 20% to about 30%, about 30% to about 40%). In yet another embodiment, the objective response rate of a patient administered a therapeutically effective amount of an anti-PD-L1 antibody, or antigen-binding portion thereof, is between about 15% to about 25%. In other embodiments, the objective response rate of a patient administered a therapeutically effective amount of a therapeutic agent against a PD-L1 antibody is at least about 15%. In other embodiments, the objective response rate of a patient administered a therapeutically effective amount of a therapeutic agent against a PD-L1 antibody is at least about 20%.

In certain embodiments, the therapies of the invention (e.g., anti-PD-L1 antibodies) are effective to increase the duration of survival of a subject. In certain embodiments, the anti-PD-L1 antibody therapy of the invention increases the duration of survival of a subject compared to standard of care therapy. In certain embodiments, the therapies of the invention increase the overall survival of the subject. In certain embodiments, the subject exhibits an overall survival of at least about 6 months, 7 months, 8 months, 9 months, 10 months, at least about 11 months, at least about 12 months, at least about 13 months, at least about 14 months at least about 15 months, at least about 16 months, at least about 17 months, at least about 18 months, at least about 19 months, at least about 20 months, at least about 21 months, at least about 22 months, at least about 23 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years after administration. In certain embodiments, the survival or duration of total survival of a subject is increased by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, or at least about 75% when compared to another subject treated with standard of care therapy alone. In other embodiments, the survival or duration of total survival of a subject is increased by at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 6 months, at least about 1 year, at least about 18 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years when compared to another subject treated with standard of care therapy alone.

In certain embodiments, the therapies of the invention are effective to increase the duration of progression-free survival of a subject. For example, progression free survival of a subject is increased by at least about 2 weeks, at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 6 months, or at least about 1 year when compared to another subject treated with standard of care therapy alone. In certain embodiments, after administration of the anti-PD-L1 antibody therapy, the subject exhibits a total response rate that is at least about 30%, 35%, 36%, 37%, 39%, 40%, 45%, or 50% compared to the response rate after administration of the standard of care therapy.

Immune checkpoint inhibitors suitable for use in the disclosed methods include anti-PD-L1 antibodies that bind to PD-L1 with high specificity and affinity, block the binding of PD-L1, and inhibit the immunosuppressive effects of the PD-1 signaling pathway. In any of the methods of treatment disclosed herein, an anti-PD-1 or anti-PD-L1 "antibody" includes an antigen-binding moiety that binds to the PD-1 or PD-L1 receptor, respectively, and exhibits functional properties in inhibiting ligand binding and upregulating the immune system similar to the functional properties of an intact antibody. In some embodiments, the anti-PD-1 antibody, anti-PD-L1 antibody, or antigen-binding portion thereof is a chimeric, humanized, or human monoclonal antibody, or portion thereof. In certain embodiments for treating a human subject, the antibody is a humanized antibody. In other embodiments for treating a human subject, the antibody is a human antibody. Antibodies of the IgG1, IgG2, IgG3 or IgG4 isotype may be used.

In certain embodiments, the anti-PD-1 antibody, anti-PD-L1 antibody, or antigen-binding portion thereof, comprises a heavy chain constant region of human IgG1 or IgG4 isotype. In certain other embodiments, the sequence of the IgG4 heavy chain constant region of the anti-PD-1 antibody, anti-PD-L1 antibody, or antigen-binding portion thereof, contains the S228P mutation that replaces a serine residue in the hinge region with a proline residue typically found at the corresponding position of an IgG1 isotype antibody. This mutation present in the monoclonal antibody prevents Fab arm exchange with endogenous IgG4 antibody while retaining low affinity for activation of the Fc receptor associated with wild-type IgG4 antibody (Wang et al, 2014Cancer Immunol res.2(9): 846-56). In other embodiments, the antibody comprises a light chain constant region that is a human kappa or lambda constant region. In other embodiments, the anti-PD-1 antibody, anti-PD-L1 antibody, or antigen-binding portion thereof is a mAb or antigen-binding portion thereof.

In WO2016022630, it has been disclosed that an anti-PD-L1 antibody has high affinity for PD-L1, can significantly inhibit the interaction of PD-L1 and PD-1 on the cell surface, and significantly promote the secretion of IL-2 and IFN-gamma by T cells.

In certain embodiments, the anti-PD-L1 antibody or fragment thereof cross-competes with 5G11 or 13C5 mab. In other embodiments, the anti-PD-L1 antibody or fragment thereof binds to the same or a similar epitope as the 5G11 or 13C5 mab. In certain embodiments, the anti-PD-L1 antibody has the same CDRs as the 5G11 or 13C5 mab.

In certain embodiments, the same epitope region of human PD-L1 is cross-competed with 5G11 or 13C5 mab. For administration to a human subject, these cross-competitive antibodies are chimeric or humanized or human antibodies. Such chimeric, humanized or human mabs may be prepared and isolated by methods well known in the art.

anti-PD-L1 antibodies useful in the disclosed inventive methods also include antigen-binding portions of the above antibodies. It is well established that the antigen binding function of an antibody can be performed by fragments of a full-length antibody. Examples of binding fragments encompassed by the term "antigen-binding portion" of an antibody include: (i) fab fragments, i.e., monovalent fragments consisting of the VL, VH, CL and CH1 domains; (ii) a F (ab ") 2 fragment, i.e. a bivalent fragment comprising 2 Fab fragments linked by a disulfide bond at the hinge region; (iii) an Fd fragment consisting of the VH and CH1 domains; and (iv) Fv fragments consisting of the VL and VH domains of a single arm of an antibody.

An anti-PD-L1 antibody suitable for use in the disclosed compositions is an antibody that binds to PD-L1 with high specificity and affinity, blocks the binding of PD-1, and inhibits the immunosuppressive effects of the PD-L1/PD-1 signaling pathway. In any of the compositions or methods disclosed herein, an anti-PD-L1 "antibody" includes an antigen-binding portion or fragment that binds to a PD-L1 ligand and exhibits functional properties similar to those of an intact antibody in inhibiting receptor binding and upregulating the immune system. In certain embodiments, the anti-PD-L1 antibody or antigen-binding portion thereof cross-competes with 5G11 or 13C5 mab for binding to human PD-L1. In other embodiments, the anti-PD-L1 antibody or antigen-binding portion thereof is a chimeric, humanized, or human monoclonal antibody or portion thereof. In certain embodiments, the antibody is a humanized antibody. In other embodiments, the antibody is a human antibody. Antibodies of the IgG1, IgG2, IgG3 or IgG4 isotype may be used.

In certain embodiments, the anti-PD-L1 antibody used in the methods may be replaced with another PD-1 or anti-PD-L1 antagonist. For example, because anti-PD-L1 antibodies prevent the interaction between PD-1 and PD-L1, thereby exerting similar effects on the signaling pathway of PD-1, anti-PD-1 antibodies may replace the use of anti-PD-L1 antibodies in the methods disclosed herein. Accordingly, in one embodiment, the invention relates to a method for treating a subject having a tumor that is primarily a mediastinal B cell lymphoma, comprising administering to the subject a therapeutically effective amount of an anti-PD-L1 antibody.

In certain embodiments, the anti-PD-L1 antibody or antigen-binding fragment thereof is a 13C5, 5G11, ch13C5-hIgG1, ch13C5-hIgG4, ch5G11-hIgG1, ch5G11-hIgG4, hu13C5-hIgG1, hu13C5-hIgG4, hu5G11-hIgG1, or hu5G11-hIgG4 monoclonal antibody or antigen-binding fragment thereof (see WO2016022630 or CN 107001463A).

The present invention provides antigen binding polypeptides or pharmaceutical compositions comprising antigen binding polypeptides for the treatment of cancer or tumors. In some embodiments, the antigen binding polypeptide is an antibody or antigen binding portion thereof that binds programmed death ligand 1(PD-L1) and/or inhibits PD-L1 activity. In some embodiments, the antigen binding polypeptide is an anti-PD-L1 antibody.

The present invention provides an isolated antibody or fragment thereof that binds PD-L1, wherein the antibody is produced by a hybridoma selected from the group consisting of the hybridomas designated herein as 13C5, 5G 11. Accordingly, the invention also includes hybridomas 13C5, 5G11, and any hybridoma that produces an antibody disclosed herein. The invention also provides isolated polynucleotides encoding the antibodies and fragments thereof provided herein. The invention also includes expression vectors comprising the isolated polynucleotides, and host cells comprising the expression vectors.

The present invention provides an anti-PD-L1 antibody comprising heavy chain Complementarity Determining Regions (CDRs) selected from the group consisting of the 13C5 or 5G11 antibodies, and light chain complementarity determining regions selected from the group consisting of the 13C5 or 5G11 antibodies. In one embodiment, the invention provides an anti-PD-L1 antibody comprising a variable heavy chain selected from a ch5G11-hIgG1, ch5G11-hIgG4, ch13C5-hIgG1, ch13C5-hIgG4 chimeric antibody, and a variable light chain selected from a ch5G11-hIgG1, ch5G11-hIgG4, ch13C5-hIgG1, ch13C5-hIgG4 chimeric antibody. In one embodiment, the invention provides an anti-PD-L1 antibody comprising a variable heavy chain selected from a humanized antibody of hu13C5-hIgG1, hu13C5-hIgG4, hu5G11-hIgG1, or hu5G11-hIgG4, and a variable light chain selected from a humanized antibody of hu13C5-hIgG1, hu13C5-hIgG4, hu5G11-hIgG1, or hu5G11-hIgG 4. Reference may be made to the description of patent documents WO2016022630 or CN 107001463A: 13C5, ch13C5-hIgG1, ch13C5-hIgG4, hu13C5-hIgG1 or hu13C5-hIgG4, wherein the sequence of HCDR1 is SYGMS, the sequence of HCDR2 is SISSGGSTYYPDSVKG, HCDR3 is GYDSGFAY, the sequence of LCDR1 is ASQSVSTSSSSFMH, LCDR2 is YASNLES, and the sequence of LCDR3 is QHSWEIPYT; 5G11, ch5G11-hIgG1, ch5G11-hIgG4, hu5G11-hIgG1 or hu5G11-hIgG4 have the sequence of HCDR1 as TYGVH, the sequence of HCDR2 as VIWRGVTTDYNAAFMS, HCDR3 as LGFYAMDY, the sequence of LCDR1 as KASQSVSNDVA, LCDR2 as YAANRYT and the sequence of LCDR3 as QQDYTSPYT.

In certain embodiments, the immunodetection point inhibitor (e.g., an anti-PD-L1 antagonist) used in the present invention is a PD-L1 Fc fusion protein.

In certain embodiments, the invention provides a method of treating a subject with a therapeutically effective amount of an anti-PD-L1 antibody, wherein the anti-PD-L1 antibody is administered alone. In some embodiments, the separate administration refers to that the anti-PD-L1 antibody may not be used in combination with, and/or not administered simultaneously with, other anti-cancer drugs. In some embodiments, the separate administration refers to the anti-PD-L1 antibody may be administered without a chemotherapeutic agent in combination with, and/or concurrently with, the chemotherapeutic agent. In some embodiments, the separate administration refers to that the anti-PD-L1 antibody may not be used in combination with, and/or not administered simultaneously with, other targeted drugs. In some embodiments, the separate administration refers to the anti-PD-L1 antibody may not be administered in combination with, and/or not administered simultaneously with, other anti-cancer antibodies. In some embodiments, the separate administration refers to that the anti-PD-L1 antibody may not be administered in combination with, and/or concurrently with, radiation therapy.

In certain embodiments, the invention provides a method of treating a disease associated with a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an anti-PD-L1 antibody, wherein the anti-PD-L1 antibody is co-administered.

In certain embodiments, the immune checkpoint inhibitor (e.g., an anti-PD-1 antibody or an anti-PD-L1 antibody) is administered in combination with one or more other anti-cancer drugs. In certain embodiments, the one or more anti-cancer agents have been administered to the subject prior to administration of the anti-PD-1 or anti-PD-L1 antibody or prior to combination with the anti-PD-1 or anti-PD-L1 antibody. In certain embodiments, the one or more anti-cancer agents are not effective in treating the cancer. In certain embodiments, the additional anti-cancer agent is any anti-cancer agent described herein or known in the art.

In certain embodiments, an anti-PD-1 antibody or an anti-PD-L1 antibody can be combined with another immunotherapy. In certain embodiments, immunotherapy involving blockade of the immune checkpoint is administered as monotherapy. In other embodiments, immunotherapy involving blockade of the immunodetection site is administered in combination with other therapies.

The therapeutic agents of the invention may be constituted in a composition, for example, a pharmaceutical composition comprising the antibody and a pharmaceutically acceptable carrier. As used herein, "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, which are physiologically compatible. In one embodiment, the carrier for the antibody-containing composition is suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal, or epidermal administration (e.g., by injection or infusion), while the carrier for the TKI-containing composition is suitable for non-parenteral (e.g., oral) administration. The pharmaceutical compositions of the present invention may include one or more pharmaceutically acceptable salts, antioxidants, aqueous and non-aqueous carriers, and/or adjuvants such as preservatives, wetting agents, emulsifying agents, and dispersing agents.

Dosage regimens are adjusted to provide the most desirable response, e.g., the maximum therapeutic response and/or the minimum adverse effects. In certain embodiments, the methods of the invention can be used with a uniform dose or a weight-based dose. In other embodiments, the anti-PD-1 antibody, anti-PD-L1 antibody, or antigen-binding portion thereof, is administered as a unitary dose. In other embodiments, the anti-PD-1 antibody, anti-PD-L1 antibody, or antigen-binding portion thereof, is administered as a weight-based dose. For administration of the anti-PD-L1 antibody (as monotherapy or in combination with another anti-cancer agent), the dose may be in the following range: from about 0.01 to about 40mg/kg, from about 0.1 to about 30mg/kg, from about 0.1 to about 20mg/kg, from about 0.1 to about 15mg/kg, from about 0.1 to about 10mg/kg, from about 1 to about 15mg/kg, from about 1 to about 20mg/kg, from about 1 to about 3mg/kg, from about 3 to about 10mg/kg, from about 3 to about 15mg/kg, from about 3 to about 20mg/kg, from about 3 to about 30mg/kg, from about 10 to about 20mg/kg, or from about 15 to about 20mg/kg of body weight, or from about 60mg to at least 2400mg, from about 90mg to at least about 1800mg, from about 120mg to at least about 1500mg, from about 300mg to at least about 9000mg, from about 600mg to at least about 900mg, from about 300mg to at least about 1200mg, from about 600mg to at least 1200mg, or from about 900mg to at least 1200 mg. For example, the dose may be about 0.1, about 1, about 2, about 3, about 5, about 6, about 9, about 10, about 15, about 20, or about 30mg/kg body weight; or about 30mg, about 60mg, about 120mg, about 150mg, about 180mg, about 300mg, about 600mg, about 900mg, about 1200mg, about 1800mg, about 2100mg, or about 2400 mg. Dosing schedules are generally designed to achieve exposures that result in sustained Receptor Occupancy (RO) (based on the typical pharmacokinetic properties of antibodies). One exemplary treatment regimen entails administration about once per week (q1w), about once every 2 weeks (q2w), about once every 3 weeks (q3w), about once every 4 weeks (q4w), about once every 1 month (q1m), about once every 3-6 months or longer. In certain embodiments, an anti-PD-L1 antibody, such as 13C5, ch13C5-hIgG1, ch13C5-hIgG4, hu13C5-hIgG1, hu13C5-hIgG4, 5G11, ch5G11-hIgG1, ch5G11-hIgG4, hu5G11-hIgG1, or hu5G11-hIgG4 monoclonal antibody, is administered to the subject about once every 2 weeks. In other embodiments, the antibody is administered about once every 3 weeks. The dosage and schedule may vary during the course of treatment. Given that IgG4 antibodies typically have a half-life of 2-3 weeks, the dosage regimen of the anti-PD-L1 antibodies of the invention comprises at least about 1 to at least about 30mg/kg body weight, at least about 3 to at least about 20mg/kg body weight, at least about 10 to at least about 15mg/kg body weight, or at least about 300 to at least about 1200mg via intravenous administration of the antibody every about 14-21 days in up to about 6 weeks or about 12 weeks of the cycle until complete response or confirmation of progressive disease. In certain embodiments, anti-PD-L1 monotherapy is administered at 3mg/kg every 2 weeks until progressive disease or unacceptable toxicity. In other embodiments, anti-PD-L1 monotherapy is administered at 1200mg every 3 weeks until progressive disease or unacceptable toxicity. In certain embodiments, the antibody treatment or any combination treatment disclosed herein lasts for at least about 1 month, at least about 3 months, at least about 6 months, at least about 9 months, at least about 1 year, at least about 18 months, at least about 24 months, at least about 3 years, at least about 5 years, or at least about 10 years.

When used in combination with other cancer agents, the dose of the anti-PD-L1 antibody can be reduced relative to a monotherapy dose. The dose of 13C5, ch13C5-hIgG1, ch13C5-hIgG4, hu13C5-hIgG1, hu13C5-hIgG4, 5G11, ch5G11-hIgG1, ch5G11-hIgG4, hu5G11-hIgG1 or hu5G11-hIgG4 monoclonal antibodies lower than the typical 20mg/kg but not less than 0.001mg/kg is a sub-therapeutic dose. Sub-therapeutic doses of anti-PD-L1 antibody used in the methods herein are above 0.001mg/kg and below 20 mg/kg. In certain embodiments, the subtherapeutic dose is from about 0.001mg/kg to about 3mg/kg, from about 0.01mg/kg to about 3mg/kg, from about 0.001mg/kg to about 10mg/kg, or from about 0.01mg/kg to about 10mg/kg of body weight. In certain embodiments, the subtherapeutic dose is at least about 0.001mg/kg, at least about 0.005mg/kg, at least about 0.01mg/kg, at least about 0.05mg/kg, at least about 0.1mg/kg, at least about 0.5mg/kg, at least about 1.0mg/kg body weight, or at least about 3.0mg/kg body weight. In certain embodiments, the sub-therapeutic unitized dose is less than about 600mg every 3 weeks, e.g., about 300mg or about 120mg every 3 weeks. In certain embodiments, 3mg/kg administration may allow sufficient exposure to result in maximal biological activity.

In certain embodiments, the dose of the anti-PD-L1 antibody or anti-PD-1 antibody is a fixed dose in a pharmaceutical composition. In other embodiments, the methods of the invention can be used in a uniform dose (a dose administered to a patient without regard to the patient's weight). For example, a uniform dose of 13C5, ch13C5-hIgG1, ch13C5-hIgG4, hu13C5-hIgG1, hu13C5-hIgG4, 5G11, ch5G11-hIgG1, ch5G11-hIgG4, hu5G11-hIgG1, or hu5G11-hIgG4 mAb may be about 1200 mg. In certain embodiments, the anti-PD-L1 antibody, or antigen-binding portion thereof, is administered at a dose of about 1200 mg. In certain embodiments, the anti-PD-L1 antibody, or antigen-binding portion thereof, is administered at a dose of about 900 mg. In certain embodiments, the anti-PD-L1 antibody, or antigen-binding portion thereof, is administered at a dose of about 600 mg. In one embodiment, 900mg of the anti-PD-L1 antibody or antigen-binding fragment is administered once every 3 weeks. In another embodiment, 1200mg of the anti-PD-L1 antibody or antigen-binding fragment is administered once every 4 weeks.

For administration of the anti-PD-L1 antibody (as monotherapy or in combination with another anti-cancer agent), the dose may be in the following range: from about 0.01 to about 20mg/kg, from about 0.1 to about 10mg/kg, from about 0.1 to about 5mg/kg, from about 3 to about 10mg/kg, from about 3 to about 15mg/kg, or from about 0.1 to about 30mg/kg of body weight or from about 80mg to at least 800mg, from about 80mg to at least about 700mg, from about 80mg to at least about 600mg, from about 80mg to at least about 500mg, from about 80mg to at least about 400mg, from about 80mg to at least about 300mg, from about 100mg to at least about 300mg, or from about 200mg to about 300 mg. For example, the dose may be about 0.1, about 0.3, about 1, about 2, about 3, about 5, or about 10mg/kg body weight, or about 0.3, about 1, about 2, about 3, or about 5mg/kg body weight; or about 80mg, about 100mg, about 160mg, about 200mg, about 240mg, about 300mg, about 320mg, about 400mg, about 500mg, about 600mg, about 700mg, or about 800 mg. Dosing schedules are generally designed to achieve exposures that result in sustained Receptor Occupancy (RO) (based on the typical pharmacokinetic properties of antibodies). An exemplary treatment regimen entails administration about once per week, about once every 2 weeks, about once every 3 weeks, about once every 4 weeks, about 1 time per month, about once every 3-6 months, or longer.

In certain embodiments, a dose of about 3mg/kg to about 30mg/kg of body weight of the subject is administered intravenously on day 1 of the cycle (D1) for about 21 days for 1 cycle until efficacy is assessed as disease progression and an intolerable toxic response is present. In certain embodiments, a body weight dose of about 3mg/kg to about 20mg/kg is administered intravenously on day 1 of the cycle (D1) for 1 cycle of about 21 days until efficacy is assessed as disease progression with intolerable toxicity. In certain embodiments, a 3mg/kg dose is received by intravenous drip on day 1 of the cycle (D1), for about 21 days for 1 cycle, until efficacy is assessed as disease progression, with intolerable toxic reactions. In certain embodiments, a dose of about 10mg/kg body weight is administered by intravenous drip on day 1 of the cycle (D1) for about 21 days for 1 cycle until therapeutic efficacy is assessed as disease progression and an intolerable toxic response occurs. In certain embodiments, a dose of about 15mg/kg body weight is administered by intravenous drip on day 1 of the cycle (D1) for about 21 days for 1 cycle until therapeutic efficacy is assessed as disease progression and an intolerable toxic response occurs. In certain embodiments, a dose of about 20mg/kg body weight is administered intravenously on day 1 of the cycle (D1) for 1 cycle for 21 days until efficacy is assessed as disease progression and an intolerable toxic response is present. In certain embodiments, a dose of about 30mg/kg body weight is administered intravenously on day 1 of the cycle (D1) for 1 cycle for 21 days until efficacy is assessed as disease progression and an intolerable toxic response is present. In certain embodiments, a body weight dose of about 3mg/kg, 10mg/kg, 15mg/kg, 20mg/kg or 30mg/kg is administered by intravenous drip on day 1 of each cycle (D1) for 1 cycle of about 2 weeks, about 3 weeks or about 4 weeks until the therapeutic effect is assessed as disease progression with intolerable toxic reactions. In certain embodiments, a single dose intravenous drip of about 1200mg is received on day 1 of the cycle (D1) for 1 cycle of about 2 weeks, about 3 weeks, or about 4 weeks, until efficacy is assessed as disease progression, with intolerable toxic reactions.

In certain embodiments, a uniform dose of about 600mg to about 1200mg is received as an intravenous drip on day 1 of the cycle (D1), for about 1 cycle for about 21 days, until efficacy is assessed as disease progression, with intolerable toxicity. In certain embodiments, a uniform dose of about 600mg to about 900mg per cycle on day 1 (D1) is administered by intravenous drip for about 21 days for 1 cycle until efficacy is assessed as disease progression and an intolerable toxic response is present. In certain embodiments, a 1200mg bolus intravenous drip is received on day 1 of the cycle (D1) for 1 cycle of about 21 days until efficacy is assessed as disease progression with intolerable toxic response. In certain embodiments, a uniform dose of about 900mg to about 1200mg per cycle on day 1 (D1) is administered by intravenous drip for about 21 days for 1 cycle until efficacy is assessed as disease progression and an intolerable toxic response is present. In certain embodiments, a 600mg bolus intravenous drip is received on day 1 of the cycle (D1) for 1 cycle for 21 days until efficacy is assessed as disease progression and an intolerable toxic response occurs. In certain embodiments, a 900mg bolus intravenous drip is received on day 1 of the cycle (D1) for 1 cycle of 21 days until efficacy is assessed as disease progression with intolerable toxic reactions. In certain embodiments, a 1200mg bolus intravenous drip is received on day 1 of the cycle (D1) for 1 cycle for 21 days until efficacy is assessed as disease progression and an intolerable toxic response occurs. In certain embodiments, a single dose intravenous drip of about 900mg is received on day 1 of the cycle (D1) for 1 cycle of about 2 weeks, about 3 weeks, or about 4 weeks, until efficacy is assessed as disease progression, with intolerable toxic reactions. In certain embodiments, a single dose intravenous drip of about 1200mg is received on day 1 of the cycle (D1) for 1 cycle of about 2 weeks, about 3 weeks, or about 4 weeks, until efficacy is assessed as disease progression, with intolerable toxic reactions.

In certain embodiments, the dose of the anti-PD-L1 antibody or anti-PD-1 antibody is a fixed dose in a pharmaceutical composition.

In certain embodiments, the dose of the anti-PD-L1 antibody or anti-PD-1 antibody is a fixed dose in a pharmaceutical composition containing a second anti-cancer agent.

The actual dosage level of one or more of the active ingredients in the pharmaceutical compositions of the invention can be varied so as to obtain an amount of the active ingredient that is effective to achieve a desired therapeutic response for a particular patient, composition, and mode of administration, without undue toxicity to the patient. The selected dosage level will depend upon a variety of pharmacokinetic factors including the activity of the particular composition of the invention employed, the route of administration, the time of administration, the rate of excretion of the particular compound being employed, the duration of the treatment, other drugs, compounds and/or materials used in conjunction with the particular composition employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts. The compositions of the present invention may be administered via one or more routes of administration using one or more of a variety of methods well known in the art. The skilled artisan will appreciate that the route and/or mode of administration will vary with the desired result.

The present invention also aims to provide a pharmaceutical composition, which is characterized in that: the pharmaceutical composition comprises an antibody and at least one or more of a buffer, an isotonic regulator, a stabilizer and/or a surfactant. In particular, the pharmaceutical composition comprises 1-150mg/ml of an anti-PD-L1 humanized monoclonal antibody (mab), 3-50mM buffer, 2-150mg/ml of an isotonic adjusting/stabilizing agent and 0.01-0.8mg/ml of a surfactant, and has a pH of about 4.5-6.8.

In some embodiments, the anti-PD-L1 humanized mab is present at a concentration of about 5 to about 150mg/ml, calculated as w/v; preferably about 10-60 mg/ml; more preferably about 10-30 mg/ml. In some embodiments, the anti-PD-L1 humanized monoclonal antibody has a mass volume concentration of about 10mg/ml, about 20mg/ml, about 30mg/ml, about 40mg/ml, about 50mg/ml, about 60mg/ml, about 70mg/ml, about 80mg/ml, about 90mg/ml, about 100mg/ml, about 110mg/ml, or about 120mg/ml, preferably about 10mg/ml, about 20mg/ml, about 30mg/ml, about 40mg/ml, about 50mg/ml, or about 60mg/ml, more preferably about 10mg/ml, about 20mg/ml, or about 30 mg/ml. In some embodiments, the anti-PD-L1 humanized monoclonal antibody mass volume concentration is about 10 mg/ml. In other embodiments, the anti-PD-L1 humanized monoclonal antibody has a mass volume concentration of about 30 mg/ml. In other embodiments, the anti-PD-L1 humanized monoclonal antibody has a mass volume concentration of about 60 mg/ml.

In some embodiments, the buffer is a histidine salt buffer. The histidine salt buffer is present at a concentration of about 5 to about 30mM, preferably about 10 to about 25mM, more preferably about 10 to about 20mM, and most preferably about 10 to about 15 mM. In some embodiments, the histidine salt buffer is about 5mM, about 10mM, about 15mM, about 20mM, about 25mM, or about 30 mM. In some embodiments, the histidine salt buffer is about 10 mM. In other embodiments, the histidine salt buffer is about 15 mM. In other embodiments, the histidine salt buffer is about 20 mM. Wherein the histidine salt buffer comprises histidine and hydrochloric acid.

In some embodiments, the isotonicity adjusting/stabilizing agent is about 20-150mg/ml sucrose, preferably about 40-100mg/ml sucrose, more preferably about 60-80mg/ml sucrose, calculated as w/v. In some embodiments, the sucrose is at a concentration of about 40mg/ml, 50mg/ml, 60mg/ml, 70mg/ml, 80mg/ml, 90mg/ml, or 100 mg/ml. In some embodiments, the sucrose is at a concentration of about 60 mg/ml. In some embodiments, the sucrose is at a concentration of about 70 mg/ml. In some embodiments, the sucrose is at a concentration of about 80 mg/ml. In some embodiments, the sucrose is at a concentration of about 90 mg/ml.

In some embodiments, the surfactant is selected from polysorbate 80, polysorbate 20, poloxamer 188; preferably polysorbate 80 or polysorbate 20; more preferably polysorbate 80. In some embodiments, the surfactant is present at a concentration of about 0.05 to about 0.6mg/ml, preferably about 0.1 to about 0.4mg/ml, and more preferably about 0.2 to about 0.3mg/ml, calculated as w/v.

In some embodiments, the surfactant is about 0.01-0.8mg/ml polysorbate 80 or polysorbate 20, calculated as w/v. In some embodiments, the surfactant is about 0.05-0.6mg/ml polysorbate 80, preferably about 0.1-0.4mg/ml polysorbate 80, more preferably about 0.2-0.3mg/ml polysorbate 80, and most preferably about 0.2mg/ml polysorbate 80. In some embodiments, the pharmaceutical composition comprises polysorbate 80 in an amount of about 0.1mg/ml, 0.2mg/ml, 0.3mg/ml, 0.4mg/ml, 0.5mg/ml, or 0.6 mg/ml; preferably, the pharmaceutical composition comprises polysorbate 80 at a level of about 0.2mg/ml, 0.3mg/ml, 0.4mg/ml or 0.5 mg/ml; more preferably, the pharmaceutical composition comprises polysorbate 80 at a level of about 0.2mg/ml, 0.3mg/ml, or 0.4 mg/ml; most preferably, the pharmaceutical composition has polysorbate 80 at a level of about 0.2 mg/ml. In some embodiments, the pharmaceutical composition comprises polysorbate 80 at a level of about 0.1 mg/ml. In other embodiments, the pharmaceutical composition comprises polysorbate 80 at a level of about 0.2 mg/ml. In some embodiments, the pharmaceutical composition comprises polysorbate 80 at a level of about 0.3 mg/ml. In other embodiments, the pharmaceutical composition comprises polysorbate 80 at a level of about 0.4 mg/ml. In some embodiments, the pharmaceutical composition comprises polysorbate 80 at a level of about 0.5 mg/ml.

In some embodiments, the pH of the aqueous solution of the pharmaceutical composition is selected from 4.0-6.8; preferably 4.5 to 6.5; more preferably 5.5 to 6.0; most preferably 5.5. In some embodiments, the pH of the aqueous pharmaceutical composition solution is about 4.5, about 4.8, about 5.0, about 5.2, about 5.4, about 5.5, about 5.6, about 5.8, or about 6.0, preferably about 5.0, about 5.2, about 5.4, about 5.5, or about 5.6, more preferably about 5.5. In some embodiments, the pH of the aqueous pharmaceutical composition solution is about 5.0. In some embodiments, the pH of the aqueous pharmaceutical composition solution is about 5.2. In some embodiments, the pH of the aqueous pharmaceutical composition solution is about 5.4. In some embodiments, the pH of the aqueous pharmaceutical composition solution is about 5.5. In some embodiments, the pH of the aqueous pharmaceutical composition solution is about 5.6. In some embodiments, the pH of the aqueous pharmaceutical composition solution is about 5.8. In some embodiments, the pH of the aqueous pharmaceutical composition solution is about 6.0.

The present invention provides isolated antibodies or fragments thereof that bind to PD-L1. The anti-PD-L1 humanized monoclonal antibody provided by the invention comprises the following amino acid sequence: a heavy chain CDR1 region having at least 80% (e.g., 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%) homology to the amino acid sequence set forth in SEQ ID NO 1 or SEQ ID NO 4; a heavy chain CDR2 region having at least 80% (e.g., 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%) homology to the amino acid sequence set forth in SEQ ID NO 2 or SEQ ID NO 5; a heavy chain CDR3 region having at least 80% (e.g., 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%) homology to the amino acid sequence set forth in SEQ ID NO 3 or SEQ ID NO 6; a light chain CDR1 region having at least 80% (e.g., 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%) homology to the amino acid sequence set forth in SEQ ID NO 7 or SEQ ID NO 10; a light chain CDR2 region having at least 80% (e.g., 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%) homology to the amino acid sequence set forth in SEQ ID NO 8 or SEQ ID NO 11; a light chain CDR3 region having at least 80% (e.g., 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%) homology to the amino acid sequence set forth in SEQ ID NO 9 or SEQ ID NO 12.

In a specific embodiment, the anti-PD-L1 humanized monoclonal antibody provided by the present invention comprises the following amino acid sequence: a heavy chain CDR1 region selected from SEQ ID NO.1 or SEQ ID NO. 4; a heavy chain CDR2 region selected from SEQ ID NO 2 or SEQ ID NO 5; a heavy chain CDR3 region selected from SEQ ID NO 3 or SEQ ID NO 6; a light chain CDR1 region selected from SEQ ID NO. 7 or SEQ ID NO. 10; a light chain CDR2 region selected from SEQ ID NO 8 or SEQ ID NO 11; a light chain CDR3 region selected from SEQ ID NO 9 or SEQ ID NO 12.

Preferably, the anti-PD-L1 humanized monoclonal antibody provided by the invention comprises the following amino acid sequence: a heavy chain variable region having at least 80% (e.g., 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) homology to the amino acid sequence set forth in SEQ ID NO 13 or SEQ ID NO 14; a light chain variable region that is at least 80% (e.g., 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) homologous to the amino acid sequence set forth in SEQ ID NO 15 or SEQ ID NO 16.

In a specific embodiment, the anti-PD-L1 humanized monoclonal antibody provided by the invention comprises the following amino acid sequence, such as a heavy chain variable region shown as SEQ ID NO: 13; the light chain variable region shown as SEQ ID NO. 15.

In another embodiment, the anti-PD-L1 humanized monoclonal antibody provided by the invention comprises the following amino acid sequence, such as the heavy chain variable region shown in SEQ ID NO: 14; the variable region of the light chain as shown in SEQ ID NO 16.

In a specific embodiment, the anti-PD-L1 humanized monoclonal antibody provided by the invention comprises a heavy chain amino acid sequence shown as SEQ ID NO.17 and a light chain amino acid sequence shown as SEQ ID NO. 18.

In another specific embodiment, the anti-PD-L1 humanized monoclonal antibody provided by the invention comprises a heavy chain amino acid sequence as shown in SEQ ID NO.19 and a light chain amino acid sequence as shown in SEQ ID NO. 20.

In another specific embodiment, the anti-PD-L1 humanized monoclonal antibody provided by the invention comprises a heavy chain amino acid sequence shown as SEQ ID NO.21 and a light chain amino acid sequence shown as SEQ ID NO. 18.

The anti-PD-L1 humanized monoclonal antibody provided by the invention can be an IgG1 antibody or an IgG4 antibody, preferably, the anti-PD-L1 humanized monoclonal antibody is an IgG1 antibody, and more preferably is a glycosylated IgG1 antibody.

In a specific embodiment of the present invention, the pharmaceutical composition comprises: (a) an anti-PD-L1 humanized mab at a mass volume concentration of about 20mg/mL, (b) sucrose at a mass volume concentration of about 70mg/mL, (c) polysorbate 80 at a mass volume concentration of about 0.1mg/mL, (d) histidine at a molar concentration of about 20mM, (e) optionally, a suitable amount of hydrochloric acid to adjust the pH of the composition to about 5.0.

In another specific embodiment of the present invention, the pharmaceutical composition comprises: (a) an anti-PD-L1 humanized mab at a mass volume concentration of about 10mg/mL, (b) sucrose at a mass volume concentration of about 80mg/mL, (c) polysorbate 80 at a mass volume concentration of about 0.2mg/mL, (d) histidine at a molar concentration of about 10mM, (e) optionally, a suitable amount of hydrochloric acid to adjust the pH of the composition to about 5.5.

In yet another embodiment of the present invention, the pharmaceutical composition comprises: (a) an anti-PD-L1 humanized mab at a mass volume concentration of about 50mg/mL, (b) sucrose at a mass volume concentration of about 80mg/mL, (c) polysorbate 80 at a mass volume concentration of about 0.3mg/mL, (d) histidine at a molar concentration of about 10mM, (e) optionally, a suitable amount of hydrochloric acid to adjust the pH of the composition to about 5.5.

In yet a more specific embodiment of the present invention, the pharmaceutical composition comprises: (a) an anti-PD-L1 humanized mab at a mass volume concentration of about 100mg/mL, (b) sucrose at a mass volume concentration of about 80mg/mL, (c) polysorbate 80 at a mass volume concentration of about 0.5mg/mL, (d) histidine at a molar concentration of about 10mM, (e) optionally, a suitable amount of hydrochloric acid to adjust the pH of the composition to about 5.5.

In yet another embodiment of the present invention, the pharmaceutical composition comprises: (a) an anti-PD-L1 humanized mab at a mass volume concentration of about 30mg/mL, (b) sucrose at a mass volume concentration of about 80mg/mL, (c) polysorbate 80 at a mass volume concentration of about 0.2mg/mL, (d) histidine at a molar concentration of about 10mM, (e) optionally, a suitable amount of hydrochloric acid to adjust the pH of the composition to about 5.5.

In yet another embodiment of the present invention, the pharmaceutical composition comprises: (a) an anti-PD-L1 humanized mab at a mass volume concentration of about 60mg/mL, (b) sucrose at a mass volume concentration of about 80mg/mL, (c) polysorbate 80 at a mass volume concentration of about 0.2mg/mL, (d) histidine at a molar concentration of about 10mM, (e) optionally, a suitable amount of hydrochloric acid to adjust the pH of the composition to about 5.5.

In yet another embodiment of the present invention, the pharmaceutical composition comprises: (a) an anti-PD-L1 humanized mab at a mass volume concentration of about 10mg/mL, (b) sucrose at a mass volume concentration of about 70mg/mL, (c) polysorbate 80 at a mass volume concentration of about 0.4mg/mL, (d) histidine at a molar concentration of about 20mM, (e) optionally, a suitable amount of acetic acid to adjust the pH of the composition to about 6.5.

In yet another embodiment of the present invention, the pharmaceutical composition comprises: (a) an anti-PD-L1 humanized mab at a mass volume concentration of about 10mg/mL, (b) sucrose at a mass volume concentration of about 80mg/mL, (c) polysorbate 80 at a mass volume concentration of about 0.2mg/mL, (d) histidine at a molar concentration of about 20mM, (e) optionally, a suitable amount of hydrochloric acid to adjust the pH of the composition to about 5.5.

In some embodiments, the pharmaceutical composition is an aqueous injection solution, including but not limited to an aqueous formulation that is not lyophilized or an aqueous formulation reconstituted from a lyophilized powder. In other embodiments, the pharmaceutical composition is a lyophilized formulation. The lyophilized preparation refers to a preparation prepared by subjecting an aqueous solution to a lyophilization process, which is a stabilization process in which a substance is first frozen, then the amount of solvent is reduced by sublimation (primary drying process) and then by desorption (secondary drying process) until the amount of solvent is a value that no longer supports biological activity or chemical reaction. The lyophilized formulation of the present invention may also be dried by other methods known in the art, such as spray drying and bubble drying.

The present invention provides formulations which do not exceed 1.1%, preferably not exceed 0.9%, more preferably not exceed 0.5% of the polymer when stored at 2-8 ℃ or 25 ℃ for at least 6 months.

The invention also provides a method for preparing the pharmaceutical composition, which comprises mixing the anti-PD-L1 humanized monoclonal antibody with other reagents, such as one or more of a buffer, an isotonic regulator/stabilizer and/or a surfactant.

The present invention also provides a method for treating a neoplastic condition in a subject comprising administering to the subject the aforementioned pharmaceutical composition.

The invention provides an article of manufacture comprising a container containing a fixed dose of an anti-PD-L1 antibody. The invention also provides for the use of an anti-PD-L1 antibody in the manufacture of a product for the treatment of cancer comprising a container comprising a fixed dose of an anti-PD-L1 antibody. In some embodiments, the container is a vial. The fixed dose is selected from about 300mg, about 600mg, about 900mg, about 1000mg, about 1200mg, about 1500mg, about 1800mg, about 2100mg, and about 2400mg of the anti-PD-L1 antibody. In some embodiments, the article of manufacture further comprises a package insert or package insert instructing the user to administer the fixed dose to a cancer patient. In some embodiments, the article of manufacture comprises 1 or more than 1 vial containing about 300mg or 600mg of anti-PD-L1 antibody. In some embodiments, the article of manufacture comprises 1 vial containing about 300mg of the anti-PD-L1 antibody. In some embodiments, the article of manufacture comprises 1 or more than 1 vial, each of which contains about 10mL of a pharmaceutical composition comprising an anti-PD-L1 antibody. In some embodiments, the article of manufacture comprises 1 or more than 1 vial, each of which contains about 20mL of the anti-PD-L1 antibody-containing pharmaceutical composition. In some embodiments, the pharmaceutical composition comprises 1-150mg/ml of the anti-PD-L1 humanized monoclonal antibody (mab), 3-50mM buffer, 2-150mg/ml of an isotonicity adjusting/stabilizing agent, and 0.01-0.8mg/ml of a surfactant, and has a pH of about 4.5-6.8. In some embodiments, the article of manufacture comprises 1 or more than 1 vial, each of which contains about 10mL of a pharmaceutical composition comprising an anti-PD-L1 antibody, wherein the pharmaceutical composition comprises 30mg/mL of the anti-PD-L1 humanized monoclonal antibody. In some embodiments, the article of manufacture comprises 1 or more than 1 vial, each of the vials containing about 20mL of a pharmaceutical composition comprising an anti-PD-L1 antibody, wherein the pharmaceutical composition comprises 30mg/mL of the anti-PD-L1 humanized monoclonal antibody. In some embodiments, the pharmaceutical composition is any one of the pharmaceutical compositions provided herein.

In some embodiments, the invention comprises:

1. a method of treating mediastinal large B-cell lymphoma in a subject, comprising administering to the subject a therapeutically effective amount of an anti-PD-L1 antibody, wherein the anti-PD-L1 antibody comprises the amino acid sequence: a heavy chain CDR1 region having at least 80% homology to the amino acid sequence set forth in SEQ ID NO.1 or SEQ ID NO. 4; a heavy chain CDR2 region having at least 80% homology to the amino acid sequence set forth in SEQ ID NO.2 or SEQ ID NO. 5; a heavy chain CDR3 region having at least 80% homology to the amino acid sequence set forth in SEQ ID NO. 3 or SEQ ID NO. 6; a light chain CDR1 region having at least 80% homology to the amino acid sequence set forth in SEQ ID NO. 7 or SEQ ID NO. 10; a light chain CDR2 region having at least 80% homology to the amino acid sequence set forth in SEQ ID NO. 8 or SEQ ID NO. 11; a light chain CDR3 region having at least 80% homology to the amino acid sequence set forth in SEQ ID NO. 9 or SEQ ID NO. 12.

2. The method of the preceding item, wherein the anti-PD-L1 antibody is administered at a dose of 1mg/kg, 3mg/kg, 10mg/kg, 15mg/kg, 20mg/kg, 25mg/kg, 30mg/kg body weight for continuous dosing.

3. The method of any one of the preceding claims, wherein the anti-PD-L1 antibody is administered in one or more unitary doses effective to treat the mediastinal large B-cell lymphoma.

4. The method of any one of the preceding claims, wherein the unitized dose is in the range of about 20mg to about 2000mg of anti-PD-L1 antibody.

5. The method of any one of the preceding claims, wherein the unitized dose is selected from about 300mg, about 600mg, about 900mg, about 1000mg, about 1200mg, about 1500mg, about 1800mg, about 2100mg, or about 2400mg of anti-PD-L1 antibody.

6. The method of any one of the preceding claims, wherein the unitized dose is selected from about 1200mg of anti-PD-L1 antibody.

7. The method of any one of the preceding claims, wherein the anti-PD-L1 antibody is administered about every week (q1w), about every 2 weeks (q1w), about every 3 weeks (q1w), or about every 4 weeks (q1 w).

8. The method according to any one of the preceding claims, wherein the anti-PD-L1 antibody is administered at a dose of about 1200mg per patient, about once every 3 weeks, for a sustained administration.

9. The method of any one of the preceding claims, wherein the patient is administered a uniform dose of the anti-PD-L1 antibody about every 3 weeks.

10. The method of any one of the preceding claims, wherein the anti-PD-L1 antibody is administered as an intravenous infusion.

11. The method of any one of the preceding claims, wherein the anti-PD-L1 antibody is administered as a 1-2 hour intravenous infusion, preferably a 1 hour intravenous infusion.

12. The method of any one of the preceding claims, wherein the anti-PD-L1 antibody is a naked antibody, an intact antibody, or an antibody fragment comprising an antigen binding region.

13. The method according to any of the preceding claims, wherein the method results in an objective response, preferably a complete response or a partial response.

14. The method of any one of the preceding claims, wherein the subject has previously received chemotherapy and/or radiation therapy.

15. The method of any one of the preceding claims, wherein the subject has received an autologous stem cell transplant following chemotherapy and/or radiation therapy.

16. The method of any one of the preceding claims, wherein disease progression resumes after the subject has achieved complete remission following chemotherapy and/or autologous stem cell transplantation.

17. The method of any one of the preceding claims, wherein the subject has failed to complete remission or failed to partial remission following chemotherapy and/or autologous stem cell transplantation.

18. The method of any one of the preceding claims, wherein the mediastinal large B-cell lymphoma has progressed following chemotherapy.

19. The method of any one of the preceding claims, wherein the mediastinal large B-cell lymphoma is a relapsed and/or refractory mediastinal large B-cell lymphoma.

20. The method of any one of the preceding claims, wherein the mediastinal large B-cell lymphoma is a relapsed or refractory mediastinal large B-cell lymphoma.

21. The method of any one of the preceding claims, wherein the mediastinal large B-cell lymphoma is relapsed.

22. The method of any one of the preceding claims, wherein the mediastinal large B-cell lymphoma is refractory.

23. The method of any one of the preceding claims, wherein the mediastinal large B-cell lymphoma treatment is treatment of relapsed or refractory mediastinal large B-cell lymphoma.

24. The method of any one of the preceding claims, wherein the relapsed or refractory mediastinal large B-cell lymphoma has not progressed during or after completion of first-line therapy.

25. The method of any one of the preceding claims, wherein the relapsed or refractory mediastinal large B-cell lymphoma has progressed following chemotherapy.

26. The method of any one of the preceding claims, wherein the mediastinal large B-cell lymphoma is metastatic.

27. The method of any one of the preceding claims, wherein the metastatic mediastinal large B-cell lymphoma has not progressed during or after completion of first line therapy.

28. The method of any one of the preceding claims, wherein the metastatic mediastinal large B-cell lymphoma has progressed following first-line treatment.

29. The method of any one of the preceding claims, wherein the metastatic mediastinal large B-cell lymphoma is a metastatic relapsed and/or refractory mediastinal large B-cell lymphoma.

30. An article of manufacture comprising a container containing a fixed dose of an anti-PD-L1 antibody, wherein the fixed dose is selected from the group consisting of about 300mg, about 600mg, about 900mg, about 1000mg, about 1200mg, about 1500mg, about 1800mg, about 2100mg, and about 2400mg of the anti-PD-L1 antibody, wherein the anti-PD-L1 antibody comprises the amino acid sequence: a heavy chain CDR1 region having at least 80% homology to the amino acid sequence set forth in SEQ ID NO.1 or SEQ ID NO. 4; a heavy chain CDR2 region having at least 80% homology to the amino acid sequence set forth in SEQ ID NO.2 or SEQ ID NO. 5; a heavy chain CDR3 region having at least 80% homology to the amino acid sequence set forth in SEQ ID NO. 3 or SEQ ID NO. 6; a light chain CDR1 region having at least 80% homology to the amino acid sequence set forth in SEQ ID NO. 7 or SEQ ID NO. 10; a light chain CDR2 region having at least 80% homology to the amino acid sequence set forth in SEQ ID NO. 8 or SEQ ID NO. 11; a light chain CDR3 region having at least 80% homology to the amino acid sequence set forth in SEQ ID NO. 9 or SEQ ID NO. 12.

31. An article of manufacture according to any preceding claim, comprising 1 or more than 1 vial containing about 300mg or 600mg of anti-PD-L1 antibody.

32. The article of manufacture of any one of the preceding claims, comprising 1 vial containing about 300mg of anti-PD-L1 antibody.

33. The article of manufacture of any one of the preceding claims, comprising 1 vial containing about 600mg of anti-PD-L1 antibody.

Definitions and explanations of terms

In order that the invention may be more readily understood, certain terms are defined. As used in this application, each of the following terms shall have the following meaning, except where otherwise explicitly provided herein. Additional definitions are set forth in the present application.

As used herein, the term "antibody" refers to a binding protein having at least one antigen binding domain. The antibodies and fragments thereof of the present invention may be whole antibodies or any fragment thereof. Thus, the antibodies and fragments of the invention include monoclonal antibodies or fragments thereof and antibody variants or fragments thereof, as well as immunoconjugates. Examples of antibody fragments include Fab fragments, Fab 'fragments, f (ab)' fragments, Fv fragments, isolated CDR regions, single chain Fv molecules (scFv), and other antibody fragments known in the art. Antibodies and fragments thereof can also include recombinant polypeptides, fusion proteins, and bispecific antibodies. The anti-PD-L1 antibodies and fragments thereof disclosed herein may be of the IgG1, IgG2, IgG3, or IgG4 isotype. The term "isotype" refers to the class of antibodies encoded by the heavy chain constant region gene. In one embodiment, the anti-PD-L1 antibodies and fragments thereof disclosed herein are of the IgG1 or IgG4 isotype. The PD-L1 antibodies and fragments thereof of the present invention may be derived from any species, including but not limited to mouse, rat, rabbit, primate, llama, and human. The PD-L1 antibody and fragments thereof can be chimeric, humanized or fully human. In one embodiment, the anti-PD-L1 antibody is an antibody produced by a mouse-derived hybridoma cell line. Thus, in one embodiment, the anti-PD-L1 antibody is a murine antibody. In another embodiment, the anti-PD-L1 antibody is a chimeric antibody. In another embodiment, the chimeric antibody is a mouse-human chimeric antibody. In another embodiment, the antibody is a humanized antibody. In another embodiment, the antibody is derived from a murine antibody and is humanized.

"humanized antibodies" are the following antibodies: the antibody comprises Complementarity Determining Regions (CDRs) derived from a non-human antibody; and framework regions and constant regions derived from human antibodies. For example, an anti-PD-L1 antibody provided herein can comprise CDRs derived from one or more murine antibodies as well as human framework and constant regions. Thus, in one embodiment, a humanized antibody provided herein binds to the same epitope on PD-L1 as a murine antibody from which the CDRs of the antibody are derived. Provided herein are exemplary humanized antibodies. Additional anti-PD-L1 antibodies or variants thereof comprising the heavy and light chain CDRs provided herein can be produced using any human framework sequence and are also included in the invention. In one embodiment, suitable for use in the invention of the framework sequences include the structure and the provision of framework sequences similar to those of the framework sequences. Additional modifications may be made in the framework regions to improve the properties of the antibodies provided herein. Such additional framework modifications may include chemical modifications; point mutations to reduce immunogenicity or to remove T cell epitopes; or reverting the mutation to a residue in the original germline sequence. In some embodiments, such modifications include those corresponding to the mutations exemplified herein, including back mutations to germline sequences. For example, in one embodiment, one or more amino acids in the human framework regions of the VH and/or VL of the humanized antibodies provided herein are back mutated to the corresponding amino acids in the parent murine antibody. For example, for the VH and VL of humanized 5G11 and humanized 13C5, several sites of the framework amino acids of the template human antibody described above were back mutated to the corresponding amino acid sequences in the mouse 5G11 and 13C5 antibodies. In one embodiment, the amino acids at positions 53 and/or 60 and/or 67 of the light chain variable region are back mutated to the corresponding amino acids found at said positions in the mouse 5G11 or 13C5 light chain variable region. In another embodiment, the amino acid at position 24 and/or 28 and/or 30 and/or 49 and/or 73 and/or 83 and/or 94 of the heavy chain variable region is back mutated to the corresponding amino acid found at said position in the mouse 5G11 or 13C5 heavy chain variable region. In one embodiment, the humanized 5G11 antibody comprises a light chain variable region wherein the amino acid at position 60 is mutated from ser(s) to asp (d) and the amino acid at position 67 is mutated from ser(s) to tyr (y); and a heavy chain variable region wherein the amino acid at position 24 is mutated from phe (f) to val (v), the amino acid at position 49 is mutated from ala (a) to gly (g), the amino acid at position 73 is mutated from thr (t) to asn (n), and the amino acid at position 83 is mutated from thr (t) to asn (n). In one embodiment, the humanized 13C5 antibody comprises a light chain variable region wherein the amino acid at position 53 is mutated from tyr (y) to lys (k); and a heavy chain variable region wherein the amino acid at position 28 is mutated from Thr (T) to Ile (I), the amino acid at position 30 is mutated from Ser (S) to Arg (R), the amino acid at position 49 is mutated from Ser (S) to Ala (A), and the amino acid at position 94 is mutated from Tyr (Y) to Asp (D). Additional or alternative back mutations can be made in the framework regions of the humanized antibodies provided herein to improve the properties of the antibody. The invention also includes humanized antibodies that bind PD-L1 and comprise framework modifications corresponding to the exemplary modifications described herein relative to any suitable framework sequence, as well as other framework modifications that otherwise improve antibody properties.

By "isolated antibody" is meant an antibody that: it is substantially free of other antibodies having different antigen specificities (e.g., an isolated antibody that specifically binds PD-1 is substantially free of antibodies that specifically bind antigens other than PD-1). However, an isolated antibody that specifically binds PD-1 may have cross-reactivity with other antigens (such as PD-1 molecules from different species). Furthermore, the isolated antibody may be substantially free of other cellular material and/or chemicals.

The term "monoclonal antibody" ("mAb") refers to a non-naturally occurring preparation of antibody molecules of single molecular composition (i.e., antibody molecules whose base sequences are substantially identical and which exhibit a single binding specificity and affinity for a particular epitope). mabs are an example of an isolated antibody. Mabs can be produced by hybridoma techniques, recombinant techniques, transgenic techniques, or other techniques known to those of skill in the art.

An "antigen-binding portion" (also referred to as an "antigen-binding fragment") of an antibody refers to one or more fragments of an antibody that retain the ability to specifically bind to an antigen bound by an intact antibody.

As used herein, the term "derived" when used in reference to a molecule or polypeptide relative to a reference antibody or other binding protein means a molecule or polypeptide that is capable of specifically binding the same epitope as the reference antibody or other binding protein.

The antibodies and antigen binding fragments thereof disclosed herein are specific for PD-L1. In one embodiment, the antibody or fragment thereof is specific for PD-L1. In one embodiment, the antibodies and fragments provided herein bind to human or primate PD-L1, but not PD-L1 from any other mammal. In another embodiment, the antibody or and fragments thereof does not bind to mouse PD-L1. The terms "human PD-L1", "hPD-L1", and "huPD-L1", and the like, are used interchangeably herein, and refer to variants or isoforms of human PD-L1 and human PD-L1. By "specific to" is meant that the antibody and fragments thereof bind PD-L1 with greater affinity than any other target.

As used herein, the term "EC 50" refers to the effective concentration, 50% of the maximal response of an antibody. As used herein, the term "IC 50" refers to the inhibitory concentration, 50% of the maximal response of an antibody. Both EC50 and IC50 may be measured by ELISA or FACS analysis or any other method known in the art.

As used herein, the term "treatment" refers to both therapeutic treatment as well as preventative or prophylactic measures. Subjects in need of treatment include those already with the disease or condition, as well as those who may have the disease or condition and whose purpose is to prevent, delay or attenuate the disease or condition.

As used herein, the term "subject" means a mammal, such as a rodent, feline, canine, and primate. Preferably, the subject according to the invention is a human.

By "administering" is meant physically introducing a composition comprising a therapeutic agent to a subject using any of a variety of methods and delivery systems known to those skilled in the art. Routes of administration of the immune checkpoint inhibitor (e.g., anti-PD-1 antibody or anti-PD-L1 antibody) include intravenous, intramuscular, subcutaneous, intraperitoneal, spinal, or other parenteral routes of administration, e.g., by injection or infusion. The phrase "parenteral administration" as used herein refers to modes of administration other than enteral and topical administration, typically by injection, and includes, but is not limited to, intravenous, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrasternal injection and infusion, and in vivo electroporation. In certain embodiments, the immune checkpoint inhibitor (e.g., an anti-PD-1 antibody or an anti-PD-L1 antibody) is administered by a non-parenteral route, and in certain embodiments, orally. Other non-parenteral routes include topical, epidermal or mucosal routes of administration, e.g., intranasally, vaginally, rectally, sublingually or topically. Administration may also be performed, for example, once, multiple times, and/or over one or more extended periods of time.

As used herein, an "adverse effect" (AE) is any adverse and often unintended or undesirable sign (including abnormal laboratory findings), symptom or disease associated with the use of medical treatment. For example, an adverse event can be associated with activation of the immune system or expansion of cells of the immune system (e.g., T cells) in response to a treatment. The medical treatment may have one or more related AEs, and each AE may have the same or different severity level. Reference to a method capable of "altering an adverse event" refers to a treatment regimen that reduces the incidence and/or severity of one or more AEs associated with the use of a different treatment regimen.

As used herein, "dosing interval" refers to the amount of time that elapses between multiple doses of a formulation disclosed herein administered to a subject. The dosing interval may thus be indicated as a range.

The term "dosing frequency" as used herein means the frequency of doses administered of a formulation disclosed herein over a given time. The frequency of administration may be indicated as the number of doses per given time, e.g. 1 time per week or 1 time per 2 weeks.

The use of the term "flat dose" refers to a dose that is administered to a patient without regard to the weight or Body Surface Area (BSA) of the patient. Thus, the unitized dose is provided as a mg/kg dose, rather than as an absolute amount of the agent (e.g., anti-PD-1 antibody). For example, a 60kg human and a 100kg human will receive the same dose of antibody (e.g., 240mg of anti-PD-1 antibody).

The use of the term "fixed dose" in relation to the compositions of the invention means that two or more different antibodies in a single composition are present in the composition in a specific (fixed) ratio to each other. In certain embodiments, the fixed dose is based on the weight of the antibody (e.g., mg). In certain embodiments, the fixed dose is based on the concentration of the antibody (e.g., mg/ml). In certain embodiments, the ratio of mg of the first antibody to mg of the second antibody is at least about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, about 1:30, about 1:40, about 1:50, about 1:60, about 1:70, about 1:80, about 1:90, about 1:100, about 1:120, about 1:140, about 1:160, about 1:180, about 1:200, about 200:1, about 180:1, about 160:1, about 140:1, about 120:1, about 100:1, about 90:1, about 80:1, about 70:1, about 60:1, about 50:1, about 40:1, about 30:1, about 20:1, about 15:1, about 10:1, about 1: 8:1, about 1:1, about 5: 7, about 1:1, about 1: 6:1, about 1:1, about, About 4:1, about 3:1, or about 2: 1. For example, a 3:1 ratio of primary and secondary antibodies may mean that the vial may contain about 240mg of primary and 80mg of secondary antibody, or about 3mg/ml of primary and 1mg/ml of secondary antibody.

The term "weight-based dose" as referred to herein refers to a dose administered to a patient that is calculated based on the weight of the patient. For example, when a patient with a weight of 60kg requires 3mg/kg of anti-PD-1 antibody and 1mg/kg of anti-CTLA-4 antibody, one can extract appropriate amounts of anti-PD-1 antibody (i.e., 180mg) and anti-CTLA-4 antibody (i.e., 60mg) at a time from a 3:1 ratio fixed dose formulation of anti-PD-1 antibody and anti-CTLA-4 antibody.

The term "immunotherapy" means the treatment of a subject having a disease or at risk of infection or of suffering from a relapse of a disease by a method that includes inducing, enhancing, suppressing or otherwise altering an immune response. By "treatment" or "therapy" of a subject is meant any type of intervention or process performed on the subject, or the administration of an active agent to a subject, with the purpose of reversing, alleviating, ameliorating, inhibiting, slowing, or preventing the onset, progression, severity, or recurrence of a symptom, complication, or condition, or biochemical indicator associated with the disease.

As used herein, "PD-L1 positive" may be used interchangeably with "at least about 1% of PD-L1 expression". In one embodiment, PD-L1 expression may be used by any method known in the art. In another embodiment, PD-L1 expression is measured by automated IHC. A PD-L1 positive tumor may thus have at least about 1%, at least about 2%, at least about 5%, at least about 10%, or at least about 20% PD-L1 expressing tumor cells as measured by automated IHC. In certain embodiments, "PD-L1 positive" means that there are at least 100 cells expressing PD-L1 on the cell surface.

"programmed death receptor-1 (PD-1)" means an immunosuppressive receptor belonging to the CD28 family. PD-1 is expressed predominantly on previously activated T cells in vivo and binds to both ligands PD-L1 and PD-L2. The term "PD-1" as used herein includes variants, homologs, and species homologs of human PD-1(hPD-1), hPD-1, and analogs having at least one common epitope with hPD-1.

"programmed death ligand-1 (PD-L1)" is one of two cell surface glycoprotein ligands for PD-1 (the other being PD-L2) that down-regulates T cell activation and cytokine secretion upon binding to PD-1.

"subject" includes any human or non-human animal. The term "non-human animal" includes, but is not limited to, vertebrates such as non-human primates, sheep, dogs, and rodents such as mice, rats, and guinea pigs. In certain embodiments, the subject is a human. The terms "subject" and "patient" are used interchangeably herein in certain contexts.

A "therapeutically effective amount" or "therapeutically effective dose" of a drug or therapeutic agent is any amount of drug that, when used alone or in combination with another therapeutic agent, protects a subject from the onset of a disease or promotes disease regression as evidenced by a reduction in the severity of disease symptoms, an increase in the frequency and duration of disease symptom-free stages, or prevention of injury or disability caused by the affliction of the disease. The ability of a therapeutic agent to promote disease regression can be evaluated using a variety of methods known to skilled practitioners, such as in human subjects during clinical trials, in animal model systems predicting efficacy for humans, or by determining the activity of the agent in an in vitro assay.

As used herein, a "sub-therapeutic dose" refers to a dose of a therapeutic compound (e.g., an antibody) that is lower than the usual or typical dose of the therapeutic compound when administered alone for the treatment of a hyperproliferative disease (e.g., cancer).

As an example, an "anti-cancer drug" promotes cancer regression in a subject or prevents further tumor growth. In certain embodiments, the therapeutically effective amount of the drug promotes cancer regression to the point of eliminating the cancer. By "promoting cancer regression" is meant that an effective amount of a drug, administered alone or in combination with an anti-neoplastic agent, results in the reduction of tumor growth or size, necrosis of the tumor, a reduction in the severity of at least one disease symptom, an increase in the frequency and duration of disease symptom-free stages, or the prevention of injury or disability resulting from the affliction of the disease. Furthermore, the terms "effective" and "effectiveness" with respect to treatment include pharmacological effectiveness and physiological safety. Pharmacological efficacy refers to the ability of a drug to promote cancer regression in a patient. Physiological safety means the level of toxicity or other adverse physiological effects (adverse effects) at the cellular, organ and/or organism level resulting from drug administration.

As an example for treating a tumor, a therapeutically effective amount of an anti-cancer agent can inhibit cell growth or tumor growth by at least about 10%, at least about 20%, at least about 40%, at least about 60%, or at least about 80% relative to an untreated subject, or, in certain embodiments, relative to a patient treated with standard of care therapy. In other embodiments of the invention, tumor regression may be observed for a period of at least about 20 days, at least about 40 days, or at least about 60 days. Despite these final measures of therapeutic effectiveness, the evaluation of immunotherapeutic drugs must also take into account "immune-related" response patterns.

By "immune-related" response pattern is meant the clinical response pattern often observed in cancer patients treated with immunotherapeutic agents that produce an anti-tumor effect by inducing a cancer-specific immune response or by altering the innate immune process. This response pattern is characterized by beneficial therapeutic effects following an initial increase in tumor burden or the appearance of new lesions, which would be classified as disease progression and would be synonymous with drug failure in the evaluation of traditional chemotherapeutic agents. Thus, proper evaluation of immunotherapeutic agents may require long-term monitoring of the effect of these agents on the target disease.

A therapeutically effective amount of a drug includes a "prophylactically effective amount," which is any amount of drug that inhibits the occurrence or recurrence of cancer when administered, alone or in combination with an anti-neoplastic agent, to a subject at risk of developing cancer (e.g., a subject with a premalignant condition) or a subject at risk of cancer recurrence. In certain embodiments, the prophylactically effective amount completely prevents the occurrence or recurrence of cancer. By "inhibiting" the occurrence or recurrence of cancer is meant reducing the likelihood of occurrence or recurrence of cancer, or completely preventing the occurrence or recurrence of cancer.

The use of alternatives (e.g., "or") should be understood to refer to either, both, or any combination of alternatives. The indefinite articles "a" or "an" as used herein shall be understood to mean "one or more" of any listed or enumerated component.

The terms "about," about, "or" consisting essentially of mean a value or composition within an acceptable error range for the particular value or composition as determined by one of ordinary skill in the art, which will depend in part on how the value or composition is measured or determined, i.e., the limitations of the measurement system. For example, "about" or "consisting essentially of can mean within 1 or more than 1 standard deviation, as practiced in the art. Alternatively, "about" or "substantially comprising" may refer to a range of up to 10% or 20% (i.e., ± 10% or ± 20%). For example, about 3mg may include any number between 2.7mg to 3.3mg (for 10%) or between 2.4mg to 3.6mg (for 20%). Furthermore, particularly with respect to biological systems or processes, the term may refer to up to an order of magnitude or up to at most 5 times the numerical value. Where a particular value or composition is provided in the application and claims, unless otherwise stated, the meaning of "about" or "consisting essentially of" should be assumed to be within an acceptable error range for that particular value or composition.

As used herein, the terms "about once per week", "about once per two weeks" or any other similar dosing interval term refer to approximations. "about once per week" may include every 7 days ± 1 day, i.e., every 6 days to every 8 days. "about once every two weeks" may include every 14 days ± 3 days, i.e., every 11 days to every 17 days. Similar approximations apply, for example, about once every 3 weeks, about once every 4 weeks, about once every 5 weeks, about once every 6 weeks, and about once every 12 weeks. In certain embodiments, a dosing interval of about once every 6 weeks or about once every 12 weeks means that a first dose may be administered on any day of the first week, and then a second dose may be administered on any day of the sixth or twelfth week, respectively. In other embodiments, a dosing interval of about once every 6 weeks or about once every 12 weeks refers to administration of a first dose on a particular day of the first week (e.g., monday) followed by administration of a second dose on the same day of the sixth or twelfth week (i.e., monday), respectively. Similar principles apply to phrases including, but not limited to, "about 1 every 2 weeks," "about 1 every month," etc. … ….

As used herein, any concentration range, percentage range, ratio range, or integer range should be understood to include the value of any integer within the recited range, and when appropriate, to include fractions thereof (such as tenths and hundredths of integers), unless otherwise indicated.

Unless otherwise stated, "about" or "approximately" in the context of the present invention means within + -5%, preferably within + -2%, more preferably within + -1% of the specified numerical range given. For example, a pH of about 5.5 means a pH of 5.5. + -. 5%, preferably a pH of 5.5. + -. 2%, more preferably a pH of 5.5. + -. 1%.

Detailed Description

The present invention is further described below with reference to specific examples, which, however, are only illustrative and not intended to limit the scope of the present invention. Likewise, the present invention is not limited to any particular preferred embodiment described herein. It will be appreciated by those skilled in the art that equivalent substitutions for the features of the invention, or corresponding modifications, may be made without departing from the scope of the invention. The reagents used in the following examples are commercially available products, and the solutions can be prepared by techniques conventional in the art, except where otherwise specified. In the examples, the anti-PD-L1 humanized monoclonal antibody was prepared as described in WO2016022630, and after affinity chromatography, an eluate containing the antibody was obtained by a conventional antibody purification method.

TABLE 1 abbreviation table

Example 1 preclinical pharmacodynamic test

This example discloses the results of a pharmacodynamic study of an anti-PD-L1 antibody in vitro and in vivo in mice.

hu5G11-hIgG1 binds to human PD-L1 protein with an EC50 of 21.3 ng/mL; the hu5G11-hIgG1 and human PD-L1 remarkably induce the secretion of IFN-gamma in a CD4+ cell MLR, the effect of the secretion is obvious dose-dependent, and the EC50 is 35.0 +/-11.3 ng/mL, which indicates that the hu5G11-hIgG1 inhibits a PD-L1/PD-1 signal channel after being combined with DC-expressed PD-L1, thereby stimulating the secretion of the IFN-gamma in a CD4+ T cell.

The hu5G11-hIgG1(15mg/kg, IP, Q2D X11) had a 91.7% tumor suppression rate against MC-38/H-11 mice subcutaneously transplanted tumors (all calculated based on median tumor volume) and significantly prolonged survival of mice with intraperitoneal inoculation of mouse MC-38/H-11 cells, with median survival >98 days, to 80% survival at the end of the experiment (D98) (p <0.01, compared to the human IgG 15mg/kg group).

Therefore, in vivo and in vitro pharmacodynamic tests prove that hu5G11-hIgG1 can prevent PD-L1 from being combined with PD-1 and B7.1 receptors on the surface of T cells, so that the T cells recover the activity, the immune response is enhanced, and the anti-tumor effect is exerted.

Example 2 preclinical toxicology testing

This example discloses the results of acute toxicity testing and long-term toxicity testing of anti-PD-L1 antibodies in animals.

The cynomolgus monkeys were used 6, and divided into 2 groups of 3, each group had both male and female. 10mg/mL of the pharmaceutical composition of the invention containing hu5G11-hIgG1 was administered at the doses of hu5G11-hIgG 1200 and 400mg/kg, respectively. The day of administration was taken as day 1 of the test. General conditions of each group of cynomolgus monkeys were observed for 14 consecutive days after administration; body weight was measured before dosing and on days 4, 9, and 14 of the experiment; measuring the food intake on the 2 nd to 3 th, 8 th to 9 th and 12 th to 13 th days of the test; measuring body temperature, II-lead electrocardiogram and blood pressure before administration, about 0-1 hour after administration and 14 days of experiment; performing hematology and blood biochemical detection on the 4 th and 14 th days of the test; all cynomolgus monkeys of each group were anesthetized on day 15 of the experiment and euthanized for anatomical observation. The test results show that: the cynomolgus monkey is injected with 200mg/kg and 400mg/kg of the product by a single intravenous injection, and no obvious abnormality is found in general observation, weight, ingestion, body temperature, electrocardiogram, blood pressure, hematology, hematobiochemistry, urine convention and general anatomy, and the Maximum Tolerated Dose (MTD) is 400 mg/kg.

The results of long-term toxicity tests show that cynomolgus monkeys intravenously injected with the pharmaceutical composition of the present invention containing hu5G11-hIgG1 for 4 weeks, were recovered from withdrawal for 4 weeks, and had a non-toxic reactive agent (NOAEL) of 200 mg/kg.

Example 3 clinical phase I protocol and safety, tolerability results

This example discloses pharmacokinetic and tolerability preliminary efficacy results for a single-center, open, dose escalation clinical phase I trial.

Clinical phase I tolerance and pharmacokinetic studies patients with advanced malignancies who were diagnosed unequivocally, failed standard therapy or lacked standard therapy were enrolled, the safety and tolerance using pharmaceutical compositions containing hu5G11-hIgG1 were observed, and the Maximum Tolerated Dose (MTD) and dose-limiting toxicity (DLT) were determined.

Primary end point: DLT; and (4) MTD.

Secondary endpoint: evaluating pharmacokinetics; the antitumor treatment effect is preliminarily examined.

Safety and tolerability results show that 1, 3, 10, 20, 30mg/kg of pharmaceutical composition containing hu5G11-hIgG1 was administered to the patients in the group. The administration was continued every 21 days for one cycle. The results show that patients currently in the group are well tolerated during continuous dosing and that the adverse effects associated with the drug do not exceed class II. The dosage can be tolerated at present; detecting the cytokines in the group with the dosage of 1-10mg/kg, and detecting the cytokines without a cytokine storm; serious adverse reactions associated with immunity were also not found for a while.

Preliminary pharmacokinetic evaluation showed that after the first intravenous drip of different doses (1, 3, 10mg/kg) of the test drug, the serum drug exposure levels of the currently enrolled patients were significantly dose dependent, showing linear pharmacokinetic profiles.

Example 4 treatment outcomes for classical Hodgkin lymphoma patients and Primary mediastinal Large B cell lymphoma patients

This example discloses preliminary efficacy assessment results for typical hodgkin lymphoma patients and primary mediastinal large B cell lymphoma patients enrolled in a clinical phase I trial.

2 subjects with classical hodgkin lymphoma, and 1 patient with primary mediastinal B cell lymphoma, had been enrolled by 2018 for 7 months, including: subject No. 3, diagnosed histopathologically as a patient with classical Hodgkin lymphoma, had lymph node metastasis following surgery and had previously undergone surgical, radiation and chemotherapy; subject No. 5, histopathologically diagnosed as a patient with primary mediastinal large B-cell lymphoma; subject No. 6, histopathologically diagnosed as a patient with classical hodgkin's lymphoma, had a tissue biopsy as the pathological tissue acquisition route, developed pleural and local chest wall metastases, and previously underwent chemotherapy.

In terms of efficacy assessment, subject 3 received a 3mg/kg body weight dose for 1 cycle of 21 days with continued dosing, with a 73% reduction in target lesions from baseline in the primary tumor assessment after 9 weeks and a 76% reduction in target lesions from baseline after 18 weeks. Subject No. 5 received a 10mg/kg body weight dose for 21 days for 1 cycle, with continuous dosing, until disease Stabilization (SD) at 27 weeks. Subject 6 received a 10mg/kg body weight dose for 21 days for 1 cycle, with continued dosing, with 55% reduction in target lesions from baseline after 9 weeks for primary tumor evaluation and 75% reduction in target lesions from baseline after 18 weeks. Also, table 2 shows the treatment progress of the already grouped cHL and rrPMBCL patients. These results suggest that the pharmaceutical composition containing hu5G11-hIgG1 significantly controlled the disease in cHL and rrPMBCL patients.

TABLE 2 treatment progression in cHL and rrPMBCL patients

Lymphoma in mm²

While the compositions and methods of this invention have been described in terms of preferred embodiments in light of the present disclosure, it will be apparent to those of skill in the art that variations may be applied to the compositions and/or methods and in the steps or in the sequence of steps of the method described herein without departing from the concept, spirit and scope of the invention.

The disclosures of all documents cited herein are incorporated by reference herein, to the extent that they provide exemplary, procedural and other details supplementary to those set forth herein.

Sequence listing

<110> Ningda Ningqing pharmaceutical industry group, Inc

CHIA TAI TIANQING PHARMACEUTICAL GROUP NANJING SHUNXIN PHARMACEUTICAL Co.,Ltd.

<120> use of anti-PD-L1 monoclonal antibody for treating cancer

<130> 2020

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Thr Tyr Gly Val His

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Val Ile Trp Arg Gly Val Thr Thr Asp Tyr Asn Ala Ala Phe Met Ser

1 5 10 15

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<211> 8

<212> PRT

<213> Mus sp.

<400> 3

Leu Gly Phe Tyr Ala Met Asp Tyr

1 5

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<211> 5

<212> PRT

<213> Mus sp.

<400> 4

Ser Tyr Gly Met Ser

1 5

<210> 5

<211> 16

<212> PRT

<213> Mus sp.

<400> 5

Ser Ile Ser Ser Gly Gly Ser Thr Tyr Tyr Pro Asp Ser Val Lys Gly

1 5 10 15

<210> 6

<211> 8

<212> PRT

<213> Mus sp.

<400> 6

Gly Tyr Asp Ser Gly Phe Ala Tyr

1 5

<210> 7

<211> 11

<212> PRT

<213> Mus sp.

<400> 7

Lys Ala Ser Gln Ser Val Ser Asn Asp Val Ala

1 5 10

<210> 8

<211> 7

<212> PRT

<213> Mus sp.

<400> 8

Tyr Ala Ala Asn Arg Tyr Thr

1 5

<210> 9

<211> 9

<212> PRT

<213> Mus sp.

<400> 9

Gln Gln Asp Tyr Thr Ser Pro Tyr Thr

1 5

<210> 10

<211> 14

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<213> Mus sp.

<400> 10

Ala Ser Gln Ser Val Ser Thr Ser Ser Ser Ser Phe Met His

1 5 10

<210> 11

<211> 7

<212> PRT

<213> Mus sp.

<400> 11

Tyr Ala Ser Asn Leu Glu Ser

1 5

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<211> 9

<212> PRT

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<400> 12

Gln His Ser Trp Glu Ile Pro Tyr Thr

1 5

<210> 13

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<213> Synthetic sequence

<400> 13

Gln Ile Thr Leu Lys Glu Ser Gly Pro Thr Leu Val Lys Pro Thr Gln

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Ser Thr Tyr

20 25 30

Gly Val His Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu Trp Leu

35 40 45

Gly Val Ile Trp Arg Gly Val Thr Thr Asp Tyr Asn Ala Ala Phe Met

50 55 60

Ser Arg Leu Thr Ile Thr Lys Asp Asn Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Asn Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Leu Gly Phe Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Leu Val

100 105 110

Thr Val Ser Ser

115

<210> 14

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<213> Synthetic sequence

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Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

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Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ile Phe Arg Ser Tyr

20 25 30

Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Ser Ile Ser Ser Gly Gly Ser Thr Tyr Tyr Pro Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Asp Cys Ala

85 90 95

Arg Gly Tyr Asp Ser Gly Phe Ala Tyr Trp Gly Gln Gly Thr Leu Val

100 105 110

Thr Val Ser Ser

115

<210> 15

<211> 107

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<213> Synthetic sequence

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Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

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Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Ser Val Ser Asn Asp

20 25 30

Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Tyr Ala Ala Asn Arg Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly

50 55 60

Ser Gly Tyr Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Asp Tyr Thr Ser Pro Tyr

85 90 95

Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

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Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Pro Gly

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Gln Arg Ala Thr Ile Thr Cys Arg Ala Ser Gln Ser Val Ser Thr Ser

20 25 30

Ser Ser Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro

35 40 45

Lys Leu Leu Ile Lys Tyr Ala Ser Asn Leu Glu Ser Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Asn

65 70 75 80

Pro Val Glu Ala Asn Asp Thr Ala Asn Tyr Tyr Cys Gln His Ser Trp

85 90 95

Glu Ile Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 17

<211> 446

<212> PRT

<213> Synthetic sequence

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Gln Ile Thr Leu Lys Glu Ser Gly Pro Thr Leu Val Lys Pro Thr Gln

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Ser Thr Tyr

20 25 30

Gly Val His Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu Trp Leu

35 40 45

Gly Val Ile Trp Arg Gly Val Thr Thr Asp Tyr Asn Ala Ala Phe Met

50 55 60

Ser Arg Leu Thr Ile Thr Lys Asp Asn Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Asn Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Leu Gly Phe Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Ala Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val

355 360 365

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

370 375 380

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

385 390 395 400

Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp

405 410 415

Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

420 425 430

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

435 440 445

<210> 18

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<212> PRT

<213> Synthetic sequence

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Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Ser Val Ser Asn Asp

20 25 30

Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Tyr Ala Ala Asn Arg Tyr Thr Gly Val Pro Asp Arg Phe Ser Gly

50 55 60

Ser Gly Tyr Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Asp Tyr Thr Ser Pro Tyr

85 90 95

Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 19

<211> 446

<212> PRT

<213> Synthetic sequence

<400> 19

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ile Phe Arg Ser Tyr

20 25 30

Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Ser Ile Ser Ser Gly Gly Ser Thr Tyr Tyr Pro Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Asp Cys Ala

85 90 95

Arg Gly Tyr Asp Ser Gly Phe Ala Tyr Trp Gly Gln Gly Thr Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Ala Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val

355 360 365

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

370 375 380

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

385 390 395 400

Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp

405 410 415

Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

420 425 430

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

435 440 445

<210> 20

<211> 218

<212> PRT

<213> Synthetic sequence

<400> 20

Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Pro Gly

1 5 10 15

Gln Arg Ala Thr Ile Thr Cys Arg Ala Ser Gln Ser Val Ser Thr Ser

20 25 30

Ser Ser Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro

35 40 45

Lys Leu Leu Ile Lys Tyr Ala Ser Asn Leu Glu Ser Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Asn

65 70 75 80

Pro Val Glu Ala Asn Asp Thr Ala Asn Tyr Tyr Cys Gln His Ser Trp

85 90 95

Glu Ile Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg

100 105 110

Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln

115 120 125

Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr

130 135 140

Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser

145 150 155 160

Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr

165 170 175

Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys

180 185 190

His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro

195 200 205

Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 21

<211> 442

<212> PRT

<213> Synthetic sequence

<400> 21

Gln Ile Thr Leu Lys Glu Ser Gly Pro Thr Leu Val Lys Pro Thr Gln

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Ser Thr Tyr

20 25 30

Gly Val His Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu Trp Leu

35 40 45

Gly Val Ile Trp Arg Gly Val Thr Thr Asp Tyr Asn Ala Ala Phe Met

50 55 60

Ser Arg Leu Thr Ile Thr Lys Asp Asn Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Asn Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Leu Gly Phe Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro

210 215 220

Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro

225 230 235 240

Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr

245 250 255

Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn

260 265 270

Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg

275 280 285

Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val

290 295 300

Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser

305 310 315 320

Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys

325 330 335

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu

340 345 350

Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe

355 360 365

Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu

370 375 380

Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe

385 390 395 400

Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly

405 410 415

Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr

420 425 430

Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly

435 440

Claims

2. The method of the preceding claim, wherein the anti-PD-L1 antibody is administered at a dose of 1mg/kg, 3mg/kg, 10mg/kg, 15mg/kg, 20mg/kg, 25mg/kg, 30mg/kg body weight for continuous dosing.

5. The method of any one of the preceding claims, wherein the anti-PD-L1 antibody is administered about every week (q1w), about every 2 weeks (q1w), about every 3 weeks (q1w), or about every 4 weeks (q1 w).

6. The method of any one of the preceding claims, wherein the anti-PD-L1 antibody is administered as an intravenous infusion.

7. The method of any one of the preceding claims, wherein the subject has previously received chemotherapy and/or radiation therapy.

8. The method of any one of the preceding claims, wherein the mediastinal large B-cell lymphoma is a relapsed and/or refractory mediastinal large B-cell lymphoma.

9. An article of manufacture comprising a container containing a fixed dose of an anti-PD-L1 antibody, wherein the fixed dose is selected from the group consisting of about 300mg, about 600mg, about 900mg, about 1000mg, about 1200mg, about 1500mg, about 1800mg, about 2100mg, and about 2400mg of the anti-PD-L1 antibody, wherein the anti-PD-L1 antibody comprises the amino acid sequence: a heavy chain CDR1 region having at least 80% homology to the amino acid sequence set forth in SEQ ID NO.1 or SEQ ID NO. 4; a heavy chain CDR2 region having at least 80% homology to the amino acid sequence set forth in SEQ ID NO.2 or SEQ ID NO. 5; a heavy chain CDR3 region having at least 80% homology to the amino acid sequence set forth in SEQ ID NO. 3 or SEQ ID NO. 6; a light chain CDR1 region having at least 80% homology to the amino acid sequence set forth in SEQ ID NO. 7 or SEQ ID NO. 10; a light chain CDR2 region having at least 80% homology to the amino acid sequence set forth in SEQ ID NO. 8 or SEQ ID NO. 11; a light chain CDR3 region having at least 80% homology to the amino acid sequence set forth in SEQ ID NO. 9 or SEQ ID NO. 12.