Use of Anti-PD-1 antibody in treatment of nasopharyngeal carcinoma

Abstract

The present invention relates to the use of an anti-PD-1 antibody or an antigen-binding fragment thereof in preparing a drug for preventing or treating a malignant cancer, and the use of a combination of an anti-PD-1 antibody or an antigen-binding fragment thereof and gemcitabine-cisplatin in preparing a drug for preventing or treating a malignant cancer. The malignant cancer is preferably nasopharyngeal carcinoma. The present invention also relates to a method for using a biomarker to predict the therapeutic effect of the anti-PD-1 antibody or the antigen-binding fragment thereof in treatment of nasopharyngeal carcinoma.

Description

Use of anti-PD-1 antibodies in the treatment of nasopharyngeal carcinoma Technical Field

[0001] The present invention relates to the use of anti-PD-1 antibodies or antigen-binding fragments thereof in the treatment of malignant tumors. Specifically, the present invention relates to the use of anti-PD-1 antibodies or antigen-binding fragments thereof in the preparation of a medicament for treating nasopharyngeal carcinoma, the use of an anti-PD-1 antibody or antigen-binding fragment thereof in combination with gemcitabine and cisplatin in the preparation of a medicament for preventing or treating malignant tumors, and a method for predicting the efficacy of anti-PD-1 antibodies or antigen-binding fragments thereof in treating nasopharyngeal carcinoma using biomarkers. Background Art

[0002] Immune escape is one of the characteristics of cancer. Ahmadzadeh, M. et al., Blood, 114:1537-44, disclosed that tumor-specific T lymphocytes are often present in the tumor microenvironment, draining lymph nodes and peripheral blood, but due to the network of immunosuppressive mechanisms in the tumor microenvironment, they are usually unable to control the progression of the tumor. + Tumor-infiltrating T lymphocytes (TILs) typically express activation-induced inhibitory receptors, including CTLA-4 and PD-1, while tumor cells frequently express immunosuppressive ligands, including PD-1 ligand 1 (PD-L1, also known as B7-H1 or CD274), which inhibits T cell activation and effector function. Among these inhibitory mechanisms, PD-1 and its ligands have emerged as important pathways that tumor cells exploit to suppress activated T cells in the tumor microenvironment.

[0003] Programmed death receptor 1 (PD-1) plays an important role in immune regulation and maintaining peripheral tolerance. PD-1 is primarily expressed in activated T and B cells and functions to inhibit lymphocyte activation, a normal peripheral tissue tolerance mechanism of the immune system that prevents immune hyperactivity. However, activated T cells infiltrating the tumor microenvironment highly express the PD-1 molecule. Inflammatory factors secreted by activated leukocytes induce tumor cells to highly express PD-1 ligands PD-L1 and PD-L2, leading to persistent activation of the PD-1 pathway on activated T cells in the tumor microenvironment. This suppresses T cell function and prevents them from killing tumor cells. Therapeutic PD-1 antibodies can block this pathway, partially restoring T cell function and enabling activated T cells to continue killing tumor cells.

[0004] Over the past decade, PD-1 / PD-L1 pathway blockade has been shown to be an effective approach for inducing durable anti-tumor responses in various cancer indications. Monoclonal antibodies (mAbs) that block the PD / PD-L1 pathway can enhance the activation and effector function of tumor-specific T cells, reduce tumor burden, and improve survival. Between 2014 and 2017, the FDA approved two anti-PD1 monoclonal antibodies (nivolumab) and three anti-PD-L1 monoclonal antibodies (atezolizumab, avelumab, and durvalumab) for the treatment of human tumors.

[0005] Nasopharyngeal carcinoma (NPC) refers to a malignant tumor that develops on the roof and side walls of the nasopharynx. Its incidence is the highest among malignant tumors of the ear, nose, and throat. A World Health Organization survey reports that 80% of NPC patients worldwide are in China. Due to its insidious onset and strong propensity to metastasize, approximately 75% of patients present at an advanced stage, with local lymph node and / or distant metastasis. While comprehensive treatment, typically focused on radiotherapy, is highly effective for early-stage NPC, recurrence or metastasis after treatment carries an extremely poor prognosis and is the primary cause of treatment failure and decreased survival. Epstein-Barr virus (EBV) infection is crucial for the development of NPC. According to the WHO classification, NPC has three histopathological types: keratinizing (type I), non-keratinizing (type II), and basosquamous cell carcinoma (type III).

[0006] However, some of these antibodies that have been marketed still have safety issues such as adverse drug reactions. Therefore, there is still a high unmet clinical need for effective therapies for malignant tumors (such as nasopharyngeal carcinoma).

[0007] Summary of the Invention

[0008] The present invention provides a use of an anti-PD-1 antibody or an antigen-binding fragment thereof in the preparation of a medicament for preventing or treating patients with malignant tumors, and a use of a combination of an anti-PD-1 antibody or an antigen-binding fragment thereof with gemcitabine and cisplatin in the preparation of a medicament for preventing or treating malignant tumors.

[0009] In another aspect, the present invention provides a method for preventing or treating malignant tumors, comprising administering to an individual in need thereof an effective amount of the anti-PD-1 antibody or antigen-binding fragment thereof of the present invention, or a combination of the anti-PD-1 antibody or antigen-binding fragment thereof with gemcitabine and cisplatin.

[0010] In another aspect, the present invention provides an anti-PD-1 antibody or antigen-binding fragment thereof, or a combination of an anti-PD-1 antibody or antigen-binding fragment thereof with gemcitabine and cisplatin, for use in treating or preventing a malignant tumor. In one or more embodiments, the malignant tumor is nasopharyngeal carcinoma.

[0011] In one or more embodiments, the malignant tumor of the present invention is recurrent or metastatic nasopharyngeal carcinoma.

[0012] In one or more embodiments, the malignant tumor of the present invention has a PD-L1 expression greater than 1% as determined by immunohistochemical staining of tumor tissue sections. As a preferred embodiment, the nasopharyngeal carcinoma of the present invention has a PD-L1 expression greater than 25% as determined by immunohistochemical staining of tumor tissue sections.

[0013] In one or more embodiments, the malignant tumor of the present invention is selected from keratinizing nasopharyngeal carcinoma and non-keratinizing nasopharyngeal carcinoma, preferably keratinizing nasopharyngeal carcinoma.

[0014] In one or more embodiments, the nasopharyngeal carcinoma described in the present invention is a nasopharyngeal carcinoma in which genomic amplification of the chromosome 11q13 region of CCND1, FGF14, FGF3 or FGF4 is not detected in peripheral blood circulating tumor DNA or tumor tissue; or the patient is a nasopharyngeal carcinoma patient in which genomic amplification of the chromosome 11q13 region of CCND1, FGF14, FGF3 or FGF4 is not detected in peripheral blood circulating tumor DNA or tumor tissue.

[0015] In one or more embodiments, the patient is a nasopharyngeal carcinoma patient whose disease is refractory to standard systemic therapy or whose disease progresses after 6 months of chemoradiotherapy.

[0016] In one or more embodiments, the nasopharyngeal carcinoma of the present invention is a nasopharyngeal carcinoma in which the number of EBV DNA copies in peripheral blood on day 28 of treatment is reduced by more than two times compared with that before administration on day 0.

[0017] In one or more embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof of the present invention comprises a light chain complementary determining region having an amino acid sequence as shown in SEQ ID NOs: 1, 2, and 3, and a heavy chain complementary determining region having an amino acid sequence as shown in SEQ ID NOs: 4, 5, and 6.

[0018] In one or more embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof of the present invention comprises a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 7, and a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 8.

[0019] In one or more embodiments, the anti-PD-1 antibody of the present invention comprises a light chain having an amino acid sequence as shown in SEQ ID NO:9, and a heavy chain having an amino acid sequence as shown in SEQ ID NO:10.

[0020] In one or more embodiments, the anti-PD-1 antibody of the present invention is selected from one or more of nivolumab, pembrolizumab, toripalimab, sintilimab, camrelizumab, tislelizumab, and cemiplimab; preferably toripalimab.

[0021] In one or more embodiments, the anti-PD-1 antibody of the present invention is a monoclonal antibody or an antigen-binding fragment thereof.

[0022] In one or more embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof described herein is administered alone.

[0023] In one or more embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof of the present invention is administered at a dosage of about 0.1 mg / kg to about 10.0 mg / kg of individual body weight, such as about 0.1 mg / kg, about 0.3 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 5 mg / kg or 10 mg / kg of individual body weight, or a fixed dose selected from about 120 mg to about 480 mg, such as a fixed dose of 120 mg, 240 mg, 360 mg or 480 mg, preferably a fixed dose of 3 mg / kg of individual body weight or a fixed dose of 240 mg.

[0024] In one or more embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof of the present invention is administered approximately once a week, once every two weeks, once every three weeks, once every four weeks, or once a month, preferably once every two weeks.

[0025] In one or more embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof of the present invention is administered at a dosage of 1 mg / kg body weight, 3 mg / kg body weight, 10 mg / kg body weight, or a fixed dose of 240 mg, or a fixed dose of 480 mg, administered once every two weeks or three weeks.

[0026] In one or more embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof of the present invention is administered in a liquid dosage form, such as an injection, via a parenteral route, such as intravenous infusion.

[0027] In one or more embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof of the present invention is administered for a period of one week, two weeks, three weeks, one month, two months, three months, four months, five months, six months or longer. Optionally, the duration of each administration cycle is the same or different, and the intervals between each administration cycle are the same or different.

[0028] In one or more embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof described herein is administered in combination with gemcitabine and cisplatin.

[0029] In one or more embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof of the present invention is administered at a dose of about 0.1 mg / kg to about 10.0 mg / kg of individual body weight, such as about 0.1 mg / kg, about 0.3 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 5 mg / kg or 10 mg / kg of individual body weight, or a fixed dose selected from about 120 mg to about 480 mg, such as a fixed dose of 120 mg, 240 mg, 360 mg or 480 mg, preferably a fixed dose of 3 mg / kg of individual body weight or a fixed dose of 240 mg; and

[0030] The single administration dose of gemcitabine is about 600 mg / m 2 to about 1400 mg / m 2 Body surface area, for example 800 mg / m 2 , 1000mg / m 2 or 1200 mg / m 2 body surface area; and

[0031] The single administration dose of cisplatin is about 40 mg / m 2 to about 120 mg / m 2 Body surface area, for example, 60 mg / m 2 , 80mg / m 2 or 100 mg / m 2 Body surface area.

[0032] In one or more embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof of the present invention is administered at a frequency of about once a week, once every two weeks, once every three weeks, once every four weeks, or once a month, preferably once every three weeks; the gemcitabine is administered at a frequency of about once a week, once every two weeks, once every three weeks, twice every three weeks, once every four weeks, or once a month, preferably twice every three weeks; and the cisplatin is administered at a frequency of about once a week, once every two weeks, once every three weeks, once every four weeks, or once a month, preferably once every three weeks.

[0033] In one or more embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof of the present invention is administered at a fixed dose of 240 mg once every three weeks; and the single administration dose of gemcitabine is about 1000 mg / m 2 body surface area, twice every three weeks; and a single administration dose of cisplatin of about 80 mg / m 2 body surface area, administered once every three weeks.

[0034] In one or more embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof, gemcitabine, and cisplatin of the present invention are administered in a liquid dosage form, such as an injection, via a parenteral route, such as intravenous infusion.

[0035] In one or more embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof, gemcitabine, and cisplatin of the present invention are administered for one week, two weeks, three weeks, one month, two months, three months, four months, five months, six months, or longer, respectively. Optionally, the duration of each administration cycle is the same or different, and the intervals between each administration cycle are the same or different.

[0036] In yet another aspect, the present invention provides a pharmaceutical combination comprising an anti-PD-1 antibody or an antigen-binding fragment thereof, gemcitabine, and cisplatin.

[0037] In one or more embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof of the present invention comprises a light chain complementarity determining region as shown in SEQ ID NOs: 1, 2 and 3, and a heavy chain complementarity determining region as shown in SEQ ID NOs: 4, 5 and 6; preferably, the anti-PD-1 antibody or antigen-binding fragment thereof comprises a light chain variable region as shown in SEQ ID NO: 7, and a heavy chain variable region as shown in SEQ ID NO: 8; preferably, the anti-PD-1 antibody comprises a light chain as shown in SEQ ID NO: 9, and a heavy chain as shown in SEQ ID NO: 10; more preferably, the anti-PD-1 antibody is toripalimab.

[0038] In another aspect, the present invention provides a reagent for detecting gene mutation or amplification of CCND1, FGF14, FGF3 or FGF4 chromosome 11q13 region in an individual's peripheral blood circulating tumor DNA and / or tumor tissue in the preparation of a kit for predicting the effect of anti-PD-1 antibodies in treating nasopharyngeal carcinoma.

[0039] In yet another aspect, the present invention provides a use of a reagent for detecting the number of EBV DNA copies in the peripheral blood of an individual in the preparation of a kit for predicting the efficacy of anti-PD-1 antibodies in treating nasopharyngeal carcinoma.

[0040] In another aspect, the present invention provides a method for predicting the effect of anti-PD-1 antibodies in treating nasopharyngeal carcinoma, which comprises detecting a gene mutation or amplification in the CCND1, FGF14, FGF3 or FGF4 chromosome 11q13 region in circulating tumor DNA in the individual's peripheral blood and / or tumor tissue before treatment, wherein the presence of a gene mutation or amplification in the CCND1, FGF14, FGF3 or FGF4 chromosome 11q13 region indicates that the malignant tumor patient is not suitable for anti-PD-1 antibodies.

[0041] In another aspect, the present invention provides a method for predicting the effect of anti-PD-1 antibodies in treating nasopharyngeal carcinoma, which comprises detecting the number of EBV DNA copies in the peripheral blood of an individual on the 28th day of treatment, wherein a decrease of more than two times in the number of EBV DNA copies in the peripheral blood indicates that the tumor patient is suitable for treatment with anti-PD-1 antibodies.

[0042] In yet another aspect, the present invention provides a detection kit comprising reagents for detecting gene mutation or amplification of CCND1, FGF14, FGF3 or FGF4 in chromosome 11q13 region in individual peripheral blood and / or tumor tissue.

[0043] In yet another aspect, the present invention provides a detection kit comprising reagents for detecting the number of EBV DNA copies in the peripheral blood of an individual.

[0044] In another aspect, the present invention provides a detection kit comprising reagents for detecting gene mutations or amplifications in the CCND1, FGF14, FGF3 or FGF4 chromosome 11q13 region in an individual's peripheral blood and / or tumor tissue, and reagents for detecting the number of EBV DNA copies in an individual's peripheral blood.

[0045] In yet another aspect, the present invention provides a kit comprising:

[0046] One or more single pharmaceutical dosage units of an anti-PD-1 antibody or an antigen-binding fragment thereof, wherein the anti-PD-1 antibody or antigen-binding fragment thereof is as described in any embodiment herein; or

[0047] One or more single pharmaceutical dosage units of an anti-PD-1 antibody or an antigen-binding fragment thereof, one or more single pharmaceutical dosage units of gemcitabine, and one or more single pharmaceutical dosage units of cisplatin; preferably, the anti-PD-1 antibody or antigen-binding fragment thereof is as described in any embodiment herein; or

[0048] One or more single pharmaceutical dosage units of a pharmaceutical combination as described in any embodiment herein. BRIEF DESCRIPTION OF THE DRAWINGS

[0049] Figure 1: Results of clinical response assessment according to RECIST v1.1. 1a: Maximum change from baseline in tumor burden for patients assessed at baseline and at least one post-treatment imaging assessment (n = 190); the length of the bar represents the maximum reduction or minimum increase in target lesions; 1b: Change in individual tumor burden over time from baseline assessment (n = 190).

[0050] Figure 2: 2a: Progression-free survival (PFS) of nasopharyngeal carcinoma patients in this study; 2b: Overall survival (OS) of nasopharyngeal carcinoma patients in this study; 2c: Duration of response (DR) of nasopharyngeal carcinoma patients in this study; 2d: Progression-free survival (PFS) of keratinizing and non-keratinizing nasopharyngeal carcinoma patients in this study; 2e: Overall survival (OS) of keratinizing and non-keratinizing nasopharyngeal carcinoma patients in this study.

[0051] Figure 3: 3a: Relationship between clinical response and tumor PD-L1 expression and TMB, PD-L1 positivity was defined as any membrane staining intensity >1% of tumor cells or immune cells using SP142 IHC staining; TMB was calculated by whole-exome sequencing of somatic mutations within coding regions; 3b: Progression-free survival (PFS) of patients with PD-L1+ vs. PD-L1-; 3c: Overall survival (OS) of patients with PD-L1+ vs. PD-L1-; 3d: Progression-free survival (PFS) of patients with the highest 10% of TMB values ​​vs. 90% of patients with the lowest TMB values; 3e: Overall survival (OS) of patients with the highest 10% of TMB values ​​vs. 90% of patients with the lowest TMB values.

[0052] Figure 4: Genetic variants and frequencies in 174 patients by whole-exome sequencing (WES).

[0053] Figure 5: 5a: Relationship between plasma EBV DNA copy number and disease stability (SD) in patients with nasopharyngeal carcinoma (n=35); 5b: Relationship between plasma EBV DNA copy number and complete remission / partial remission (CR / PR) in patients with nasopharyngeal carcinoma (n=34); 5c: Relationship between plasma EBV DNA copy number and disease progression (PD) in patients with nasopharyngeal carcinoma (n=80).

[0054] Figure 6: 6a: PFS assessed by independent review committee according to RECIST v1.1 (intention-to-treat population); 6b: PFS assessed by investigator according to RECIST v1.1 (intention-to-treat population); 6c: Subgroup treatment effect (progression-free survival).

[0055] Figure 7: Overall survival (OS) (intention-to-treat population).

[0056] Figure 8: Duration of response as assessed by an independent review committee according to RECIST v1.1.

[0057] In the figure, “chemotherapy” refers to the administration of gemcitabine and cisplatin. DETAILED DESCRIPTION

[0058] The present invention relates to methods for treating malignant tumors. The methods of the present invention comprise administering an anti-PD-1 antibody or an antigen-binding fragment thereof to a patient in need thereof. The malignant tumor described herein is nasopharyngeal carcinoma. The present invention also relates to methods for using biomarkers to predict the efficacy of anti-PD-1 antibodies in treating malignant tumors, particularly nasopharyngeal carcinoma.

[0059] the term

[0060] In order to make the present invention easier to understand, certain technical terms are specifically defined below. Unless otherwise explicitly stated elsewhere in this document, the technical terms used herein have the meanings commonly understood by those of ordinary skill in the art to which the present invention belongs.

[0061] "Administering," "giving," and "treating" refer to introducing a composition comprising a therapeutic agent into a subject using any of a variety of methods or delivery systems known to those skilled in the art. Routes of administration of anti-PD-1 antibodies include intravenous, intramuscular, subcutaneous, peritoneal, spinal, or other parenteral routes of administration, such as injection or infusion. "Parenteral administration" refers to administration other than enteral or topical administration, typically by injection, including but not limited to intravenous, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intracortical, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, intradural, and intrasternal injection and infusion, as well as in vivo electroporation.

[0062] An "adverse effect" (AE) as described herein is any unfavorable and generally unintended or undesirable sign, symptom, or disease associated with the use of a medical treatment. For example, an adverse effect may be associated with activation of the immune system or expansion of immune system cells in response to treatment. A medical treatment may have one or more associated AEs, and each AE may have the same or different levels of severity.

[0063] "Tumor burden" refers to the total amount of tumor material distributed throughout the body. Tumor burden refers to the total number of cancer cells or the total size of a tumor throughout the body. Tumor burden can be determined by a variety of methods known in the art, such as measuring the size of a tumor after it has been removed from a subject using calipers, or while in vivo using imaging techniques such as ultrasound, bone scans, computed tomography (CT), or magnetic resonance imaging (MRI) scans.

[0064] The term "tumor size" refers to the overall size of a tumor, which can be measured as the length and width of the tumor. Tumor size can be determined by a variety of methods known in the art, such as measuring the size of the tumor using calipers after removal from the subject, or using imaging techniques (such as bone scans, ultrasound, CT or MRI scans) while in vivo.

[0065] The terms "subject," "individual," and "object" include any organism, preferably an animal, more preferably a mammal (e.g., rat, mouse, dog, cat, rabbit, etc.), and most preferably a human. The terms "subject" and "patient" are used interchangeably herein.

[0066] As used herein, "antibody" refers to any form of antibody that can achieve the desired biological activity or binding activity. Therefore, it is used in the broadest sense, but is not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies, humanized full-length human antibodies, chimeric antibodies, and camelid-derived single-domain antibodies. An "antibody" specifically binds to an antigen and comprises at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each heavy chain comprises a heavy chain variable region (VH) and a heavy chain constant region, the heavy chain constant region comprising three constant domains, CH1, CH2, and CH3. Each light chain comprises a light chain variable region (VL) and a light chain constant region, the light chain constant region comprising one constant domain, CL. The VH and VL regions can be further subdivided into hypervariable regions, known as complementarity determining regions (CDRs), which are interspersed with more conserved regions known as framework regions (FRs). Generally speaking, from N-terminus to C-terminus, both the light and heavy chain variable domains comprise FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. Amino acids are typically assigned to each domain according to the following definitions: Sequences of Proteins of Immunological Interest, Kabat et al.; National Institutes of Health, Bethesda, Md.; 5th Edition; NIH Publication No. 91-3242 (1991); Kabat (1978) Adv. Prot. Chem. 32: 1-75; Kabat et al., (1977) J. Biol. Chem. 252: 6609-6616; Chothia et al., (1987) J Mol. Biol. 196: 901-917 or Chothia et al., (1989) Nature 341: 878-883.

[0067] The carboxyl-terminal portion of the heavy chain defines the constant region primarily responsible for effector function. Human light chains are typically classified as kappa and lambda chains. Human heavy chains are typically classified as μ, δ, γ, α, or ε, and define the antibody isotype as IgM, IgD, IgG, IgA, and IgE, respectively. IgG subclasses are well known to those skilled in the art and include, but are not limited to, IgG1, IgG2, IgG, and IgG4.

[0068] The term "antibody" includes: naturally occurring and non-naturally occurring Abs; monoclonal and polyclonal Abs; chimeric and humanized Abs; human or non-human Abs; fully synthetic Abs; and single-chain Abs. Non-human Abs can be humanized by recombinant methods to reduce their immunogenicity in humans.

[0069] Unless otherwise expressly indicated, "antibody fragments" or "antigen-binding fragments" as used herein refer to antigen-binding fragments of antibodies, i.e., antibody fragments that retain the ability of a full-length antibody to specifically bind to an antigen, such as fragments that retain one or more CDR regions. Examples of antigen-binding fragments include, but are not limited to, Fab, Fab', F(ab')2, and Fv fragments; diabodies; linear antibodies; single-chain antibody molecules; nanobodies; and multispecific antibodies formed from antibody fragments.

[0070] "Chimeric antibodies" refer to antibodies and fragments thereof in which a portion of the heavy chain and / or light chain is identical or homologous to the corresponding sequence in antibodies derived from a particular species (such as human) or belonging to a particular antibody class or subclass, while the remainder of the chain is identical or homologous to the corresponding sequence in antibodies derived from another species (such as mouse) or belonging to another antibody class or subclass, as long as it exhibits the desired biological activity.

[0071] A "human antibody" refers to an antibody that comprises only human immunoglobulin sequences. If the human antibody is produced in a mouse, mouse cell, or hybridoma derived from a mouse cell, it may contain murine carbohydrate chains. Similarly, a "mouse antibody" or "rat antibody" refers to an antibody that comprises only mouse or rat immunoglobulin sequences, respectively.

[0072] "Humanized antibody" refers to an antibody form containing sequences from non-human (e.g., murine) antibodies as well as human antibodies. Such antibodies contain minimal sequences derived from non-human immunoglobulins. Typically, a humanized antibody will comprise substantially all of at least one and typically two variable domains, wherein all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin, and all or substantially all of the FR regions are those of a human immunoglobulin. The humanized antibody optionally further comprises at least a portion of an immunoglobulin constant region (Fc), typically a human immunoglobulin constant region.

[0073] The term "nasopharyngeal carcinoma" refers to a malignant tumor that occurs in the nasopharynx or upper throat. Common clinical symptoms include nasal congestion, blood in mucus, stuffy ears, hearing loss, diplopia, and headache. Epstein-Barr virus (EBV) infection is crucial for the development of NPC. According to the WHO classification, there are three histopathological types of nasopharyngeal carcinoma: keratinizing type (type I), non-keratinizing type (type II), and basosquamous cell carcinoma (type III). Non-keratinizing nasopharyngeal carcinoma is closely related to EBV, has a higher response to radiotherapy, and has a higher overall survival rate.

[0074] The term "immunotherapy" refers to the treatment of a subject who has a disease or is at risk of contracting or suffering from a recurrence of a disease by methods that include inducing, enhancing, suppressing, or otherwise modifying an immune response. "Treatment" or "therapy" of a subject refers to any type of intervention or procedure performed on a subject, or the administration of an active agent to a subject, with the intent to reverse, alleviate, ameliorate, slow, or prevent the onset, progression, severity, or recurrence of symptoms, complications, or conditions, or biochemical markers associated with a disease.

[0075] "Programmed death receptor-1 (PD-1)" refers to an immunoinhibitory receptor belonging to the CD28 family. PD-1 is primarily expressed on previously activated T cells in vivo and binds to two ligands, PD-L1 and PD-L2. The term "PD-1" as used herein includes human PD-1 (hPD-1), variants, isoforms, and species homologs of hPD-1, as well as analogs that share at least one common epitope with hPD-1.

[0076] A "therapeutically effective amount" or "therapeutically effective dose" of a drug or therapeutic agent is any amount of the drug that, when used alone or in combination with another therapeutic agent, protects a subject from the onset of 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 periods, or the prevention of impairment or disability resulting from 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 those skilled in the art, such as in human subjects during clinical trials, in animal model systems predictive of human efficacy, or by measuring the activity of the agent in in vitro assays.

[0077] A therapeutically effective amount of a drug includes a "prophylactically effective amount," ie, any amount of a drug that, when administered alone or in combination with an anti-tumor agent to a subject at risk of developing cancer or to a subject who has had a recurrence of cancer, inhibits the development or recurrence of cancer.

[0078] "Biotherapeutic agent" refers to a biological molecule, such as an antibody or fusion protein, that blocks ligand / receptor signaling in any biological pathway that supports tumor maintenance and / or growth or suppresses anti-tumor immune responses.

[0079] Unless expressly indicated otherwise, "CDR" as used herein refers to the complementarity determining regions of immunoglobulin variable regions as defined using the Kabat numbering system.

[0080] "Therapeutic anti-PD-1 monoclonal antibodies" refer to antibodies that specifically bind to a specific mature form of PD-1 expressed on the surface of certain mammalian cells. Mature PD-1 lacks a pre-secretory leader sequence, or leader peptide. The terms "PD-1" and "mature PD-1" are used interchangeably herein and, unless otherwise explicitly defined or clearly evident from the context, should be understood to refer to the same molecule.

[0081] As described herein, therapeutic anti-human PD-1 antibodies or anti-hPD-1 antibodies refer to monoclonal antibodies that specifically bind to mature human PD-1.

[0082] As used herein, "framework region" or "FR" refers to an immunoglobulin variable region excluding the CDR regions.

[0083] An "isolated antibody or antigen-binding fragment thereof" refers to a purified state and in which case the designated molecule is substantially free of other biomolecules, such as nucleic acids, proteins, lipids, carbohydrates, or other materials (such as cell debris or growth medium).

[0084] "Patient," "patient," or "subject" refers to any single human, typically a mammal, including humans and other mammals such as horses, cows, dogs, or cats, who is in need of medical treatment or is participating in a clinical trial, epidemiological study, or serving as a control.

[0085] The "RECIST 1.1 efficacy criteria" described herein refer to the definitions of target damage or non-target damage described in Eisenhauver et al., EA et al., Eur. J Cancer 45:228-247 (2009) based on the context of the measured response. Before immunotherapy, it was the most commonly used standard for evaluating the efficacy of solid tumors. However, with the advent of the immune era, many problems that had not previously appeared in tumor evaluation have emerged. Therefore, based on the emerging phenomena caused by immunotherapy itself, in 2016, the RECIST Working Group revised the existing "RECIST v.1.1" and proposed a new judgment standard, namely the "irRECIST standard" described herein, which aims to better evaluate the efficacy of immunotherapy drugs.

[0086] The Eastern Cooperative Oncology Group (ECOG) performance status score (ECOG) is an indicator of a patient's general health and ability to tolerate treatment based on their physical strength. The ECOG performance status score is 0, 1, 2, 3, 4, or 5. A score of 0 indicates completely normal activity, no difference from pre-onset activity. A score of 1 indicates the patient can move freely and engage in light physical activity, including general housework or office work, but cannot engage in heavy physical activity.

[0087] "Sustained response" refers to a persistent therapeutic effect after cessation of treatment with a therapeutic agent or combination therapy described herein. In some embodiments, the sustained response has a duration that is at least the same as the duration of treatment or at least 1.5, 2.0, 2.5, or 3 times the duration of treatment.

[0088] "Tissue section" refers to a single portion or piece of a tissue sample, such as a thin slice of tissue cut from a sample of normal tissue or a tumor.

[0089] As used herein, "treating" cancer refers to administering a treatment regimen described herein (e.g., administering an anti-PD-1 antibody) to a subject having or diagnosed with cancer to achieve at least one positive therapeutic effect (e.g., a decrease in the number of cancer cells, a decrease in tumor volume, a decrease in the rate of cancer cell infiltration into peripheral organs, or a decrease in the rate of tumor metastasis or tumor growth). Positive therapeutic effects in cancer can be measured in a variety of ways (see WA Weber, J. Nucl. Med., 50:1S-10S (2009)). For example, with respect to tumor growth inhibition, according to NCI criteria, a T / C ≤ 42% is the minimum level of anti-tumor activity. T / C (%) = median treated tumor volume / median control tumor volume × 100. PFS (also called "time to tumor progression") refers to the length of time during and after treatment that the cancer does not grow, and includes the amount of time a patient experiences a CR or PR and the amount of time a patient experiences SD. DFS refers to the length of time a patient remains disease-free during and after treatment. OS refers to the extension of life expectancy compared to an initially or untreated individual or patient. The treatment regimen of the combination of the present invention that is effective in treating a cancer patient may vary according to a variety of factors, such as the patient's disease state, age, weight, and the ability of the therapy to stimulate an anti-cancer response in the subject. Although embodiments of the present invention may not achieve an effective positive therapeutic effect in every subject, they should be effective and achieve a positive therapeutic effect in a statistically significant number of subjects.

[0090] The terms "administration method" and "dosage regimen" are used interchangeably to refer to the dosage and timing of each therapeutic agent in the combination of the present invention.

[0091] The term "immunohistochemistry (IHC)" refers to a method for determining the location, qualitative, and relative quantitative analysis of antigens (peptides and proteins) within tissue cells by using the principle of specific binding between antigens and antibodies to develop color with a colorant (fluorescein, enzyme, metal ion, isotope) labeled with the antibody through a chemical reaction. In some embodiments of the present invention, prior to treatment with an anti-PD-1 antibody, a PD-L1 test is performed on a tumor tissue sample from a subject using Roche's anti-human PD-L1 antibody SP142 (Cat No: M4422) for staining. In some embodiments, a membrane staining intensity of ≥1% of tumor cells is defined as PD-L1 positive.

[0092] As used herein, the term "cancer" or "malignancy" refers to a broad range of diseases characterized by the uncontrolled growth of abnormal cells in the body. Unregulated cell division, proliferation, and growth lead to the formation of malignant tumors, which invade adjacent tissues and may also metastasize to distant parts of the body via the lymphatic system or bloodstream. Examples of cancers suitable for treatment or prevention using the methods, medicaments, and kits of the present invention include, but are not limited to, carcinomas, lymphomas, leukemias, blastomas, and sarcomas. More specific examples of cancer include squamous cell carcinoma, myeloma, small cell lung cancer, non-small cell lung cancer, glioma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, acute myeloid leukemia, multiple myeloma, gastrointestinal cancer, kidney cancer, ovarian cancer, liver cancer, lymphoblastic leukemia, lymphocytic leukemia, colorectal cancer, endometrial cancer, kidney cancer, prostate cancer, thyroid cancer, melanoma, chondrosarcoma, neuroblastoma, pancreatic cancer, glioblastoma multiforme, nasopharyngeal cancer, cervical cancer, brain cancer, stomach cancer, bladder cancer, hepatoma, breast cancer, colon cancer, and head and neck cancer.

[0093] As used herein, the term "tumor mutation burden (TMB)" refers to the total number of somatic gene coding errors, base substitutions, gene insertions, or deletions detected per million bases. In some embodiments of the present invention, tumor mutation burden (TMB) is estimated by analyzing somatic mutations, including coding base substitutions and megabase insertions of the studied panel sequences.

[0094] In the following paragraphs, various aspects of the invention are described in further detail.

[0095] Anti-PD-1 antibodies

[0096] As used herein, "PD-1 antibody" refers to any chemical compound or biological molecule that binds to the PD-1 receptor, blocks the binding of PD-L1 expressed on cancer cells to PD-1 expressed on immune cells (T, B, NK cells), and preferably also blocks the binding of PD-L2 expressed on cancer cells to PD-1 expressed on immune cells. Alternative terms or synonyms for PD-1 and its ligands include: PDCD1, PD1, CD279, and SLEB2 for PD-1; PDCD1L1, PDL1, B7-H1, B7H1, B7-4, CD274, and B7-H for PD-L1; and PDCD1L2, PDL2, B7-DC, and CD273 for PD-L2. In any of the methods, medicaments, and uses of the present invention for treating human subjects, the PD-1 antibody blocks the binding of human PD-L1 to human PD-1, and preferably blocks the binding of both human PD-L1 and PD-L2 to human PD1. The amino acid sequence of human PD-1 can be found at NCBI Locus Accession No. NP_005009. The amino acid sequences of human PD-L1 and PD-L2 can be found at NCBI Locus Accession No. NP_054862 and NP_079515, respectively.

[0097] Herein, when referring to an "anti-PD-1 antibody," the term includes antigen-binding fragments thereof unless otherwise indicated or described.

[0098] The anti-PD-1 antibodies suitable for any of the uses, therapies, drugs and kits described herein bind to PD-1 with high specificity and affinity, block the binding of PD-L1 / 2 to PD-1, and inhibit PD-1 signal transduction, thereby achieving an immunosuppressive effect. In any of the uses, therapies, drugs and kits disclosed herein, the anti-PD-1 antibodies include the full-length antibodies themselves, as well as antigen-binding portions or fragments that bind to the PD-1 receptor and exhibit functional properties similar to those of the complete Ab in inhibiting ligand binding and upregulating the immune system. In some embodiments, the anti-PD-1 antibody or its antigen-binding fragment is an anti-PD-1 antibody or its antigen-binding fragment that cross-competes with toripalimab for binding to human PD-1. In other embodiments, the anti-PD-1 antibody or its antigen-binding fragment is a chimeric, humanized or human Ab or its antigen-binding fragment. In certain embodiments for treating human individuals, the Ab is a humanized Ab.

[0099] In some embodiments, the anti-PD-1 antibody for any of the uses, therapies, medicaments, and kits described herein comprises a monoclonal antibody (mAb) or an antigen-binding fragment thereof that specifically binds to PD-1, and preferably specifically binds to human PD-1. The mAb can be a human antibody, a humanized antibody, or a chimeric antibody, and can include a human constant region. In some embodiments, the constant region is selected from the group consisting of human IgG1, IgG2, IgG3, and IgG4 constant regions; preferably, the anti-PD-1 antibody or antigen-binding fragment thereof for any of the uses, therapies, medicaments, and kits described herein comprises a heavy chain constant region of the human IgG1 or IgG4 isotype, more preferably a human IgG4 constant region. In some embodiments, the sequence of the IgG4 heavy chain constant region of the anti-PD-1 antibody or antigen-binding fragment thereof comprises an S228P mutation, which replaces a serine residue in the hinge region with a proline residue that is typically present at the corresponding position in antibodies of the IgG1 isotype.

[0100] Preferably, in any embodiment of the use, therapy, drug and kit described in the present invention, the PD-1 antibody is a monoclonal antibody or an antigen-binding fragment thereof, and its light chain CDRs are the amino acids shown in SEQ ID NOs: 1, 2 and 3, and the heavy chain CDRs are the amino acids shown in SEQ ID NOs: 4, 5 and 6.

[0101] More preferably, in any embodiment of the use, therapy, medicine and kit described in the present invention, the PD-1 antibody is a monoclonal antibody that specifically binds to human PD-1 and comprises: (a) a light chain variable region comprising SEQ ID NO: 7, and (b) a heavy chain variable region comprising SEQ ID NO: 8.

[0102] Further preferably, in any embodiment of the use, therapy, medicine and kit described in the present invention, the PD-1 antibody is a monoclonal antibody that specifically binds to human PD-1 and comprises: (a) a light chain comprising SEQ ID NO: 9, and (b) a heavy chain comprising SEQ ID NO: 10.

[0103] Table A below provides the amino acid sequence numbers of the light chain CDRs and heavy chain CDRs of exemplary anti-PD-1 antibody mAbs for use in the uses, therapies, medicaments, and kits described herein:

[0104] Table A: Light and heavy chain CDRs of exemplary anti-human PD-1 antibodies

[0105] LCDR1SEQ ID NO: 1LCDR2SEQ ID NO: 2LCDR3SEQ ID NO: 3HCDR1SEQ ID NO: 4HCDR2SEQ ID NO: 5HCDR3SEQ ID NO: 6

[0106] Examples of anti-PD-1 antibodies that bind to human PD-1 and can be used in the uses, therapies, medicaments, and kits described herein are described in WO2014206107. Human PD-1 mAbs that can be used as anti-PD-1 antibodies in the uses, therapies, medicaments, and kits described herein include any of the anti-PD-1 antibodies described in WO2014206107, including Toripalimab (a humanized IgG4 mAb having a structure described in WHO Drug Information (Vol. 32, No. 2, pp. 372-373 (2018)) and comprising the light and heavy chain amino acid sequences set forth in SEQ ID NOs: 9 and 10). In preferred embodiments, the anti-PD-1 antibody that can be used in any of the uses, therapies, medicaments, and kits described herein is selected from humanized antibodies 38, 39, 41, and 48 described in WO2014206107. In particularly preferred embodiments, the anti-PD-1 antibody that can be used in any of the uses, therapies, medicaments, and kits described herein is Toripalimab.

[0107] Anti-PD-1 antibodies that can be used in any of the uses, therapies, drugs, and kits described in the present invention also include Nivolumab and Pembrolizumab approved by the FDA.

[0108] In certain embodiments, the anti-PD-1 antibodies that can be used in any of the uses, therapies, drugs, and kits described herein also include anti-PD-L1 monoclonal antibodies that specifically bind to PD-L1 to block the binding of PD-L1 to PD-1, such as nivolumab, pembrolizumab, toripalimab, sintilimab, camrelizumab, tislelizumab, and cemiplimab.

[0109] As used herein, "PD-L1" expression or "PD-L2" expression refers to any detectable expression level of a specific PD-L protein on the surface of a cell or a specific PD-L mRNA within a cell or tissue. PD-L protein expression can be detected using diagnostic PD-L antibodies in IHC analysis of tumor tissue sections or by flow cytometry. Alternatively, PD-L protein expression by tumor cells can be detected by PET imaging using a binding agent that specifically binds to the desired PD-L target (such as PD-L1 or PD-L2).

[0110] For methods for quantifying PD-L1 protein expression in IHC analysis of tumor tissue sections, see, but are not limited to, Thompson, RH et al., PNAS 101(49): 17174-17179 (2004); Taube, JM et al., Sci Transl Med 4, 127ra37 (2012); and Toplian, SL et al., New Eng. J. Med. 366(26): 2443-2454 (2012).

[0111] One approach uses a simple binary endpoint of positive or negative PD-L1 expression, where a positive result is defined as the percentage of tumor cells showing histological evidence of cell surface membrane staining. Positive PD-L1 expression is defined as a count of greater than 1% of total tumor cells in a tumor tissue section.

[0112] In another approach, PD-L1 expression in tumor tissue sections is quantified in tumor cells and in infiltrating immune cells. The percentage of tumor cells and infiltrating immune cells exhibiting membrane staining is quantified separately as ≤1%, 1% to 50%, and then 50% to 100%. For tumor cells, PD-L1 expression is counted as negative if the score is ≤1% and as positive if the score is >1%.

[0113] In some embodiments, the level of PD-L1 expression by malignant cells and / or by infiltrating immune cells within a tumor is determined to be "overexpressed" or "elevated" based on comparison to the level of PD-L1 expression by an appropriate control. For example, the control PD-L1 protein or mRNA expression level can be the level quantified in non-malignant cells of the same type or in sections from matched normal tissue.

[0114] Gemcitabine

[0115] Gemcitabine is a new cytosine nucleoside derivative with the structure shown below:

[0116]

[0117] Like cytarabine, gemcitabine is activated by deoxycytidine kinase and metabolized by cytidine deaminase upon entry into the human body. Difluorodeoxycytidine, the primary metabolite of gemcitabine, is incorporated into DNA intracellularly, primarily acting during the G1 / S phase. However, difluorodeoxycytidine differs from cytarabine in that it not only incorporates into DNA but also inhibits ribonucleotide reductase, leading to a decrease in intracellular deoxynucleoside triphosphates. Another difference from cytarabine is that gemcitabine inhibits deoxycytidine deaminase, reducing the degradation of intracellular metabolites, resulting in a self-enhancing effect.

[0118] In some embodiments of the present invention, gemcitabine may also refer to a composition comprising a therapeutically effective amount of the compound represented by the above structural formula, a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0119] Cisplatin

[0120] Cisplatin is a platinum-containing anticancer drug, namely cis-dichlorodiammineplatinum, which is an orange-yellow or yellow crystalline powder. It is slightly soluble in water and easily soluble in dimethylformamide. It can gradually convert into the trans form and hydrolyze in aqueous solution. Cisplatin is a compound with the following structure:

[0121]

[0122] In some embodiments of the present invention, cisplatin may also refer to a composition comprising a therapeutically effective amount of the compound represented by the above formula, a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0123] Drug combinations

[0124] The present invention also provides a drug combination comprising the anti-PD-1 antibody described herein, gemcitabine and cisplatin. In this drug combination, the anti-PD-1 antibody, gemcitabine and cisplatin can be provided in the form of a mixture of the three (i.e., in the form of a pharmaceutical composition), or provided in the form of a mixture of any two and another independent preparation, or each is provided in the form of an independent preparation. In some embodiments, the drug combination contains a three-week dosage, including 1 dose of the anti-PD-1 antibody described herein, 2 doses of gemcitabine and 1 dose of cisplatin. When present in the form of an independent preparation, each preparation contains a pharmaceutically acceptable carrier in addition to the active ingredient.

[0125] In some embodiments, the anti-PD-1 antibody of the present invention may be as described in any embodiment herein, more preferably an antibody having a light chain CDR with amino acids shown in SEQ ID NOs: 1, 2, and 3, and a heavy chain CDR with amino acids shown in SEQ ID NOs: 4, 5, and 6, more preferably a monoclonal antibody comprising a light chain variable region shown in SEQ ID NO: 7 and a heavy chain variable region shown in SEQ ID NO: 8, more preferably a monoclonal antibody comprising a light chain shown in SEQ ID NO: 9 and a heavy chain shown in SEQ ID NO: 10, more preferably humanized antibodies 38, 39, 41, and 48 described in WO2014206107, and most preferably toripalimab.

[0126] The pharmaceutical combinations of the present invention may also include one or more additional therapeutic agents. The additional therapeutic agents may be, for example, chemotherapeutic agents, biotherapeutic agents, immunogenic agents (e.g., attenuated cancer cells, tumor antigens, antigen-presenting cells (such as dendritic cells pulsed with tumor-derived antigens or nucleic acids), immunostimulatory cytokines (e.g., IL-2, IFN-γ, GM-CSF), and cells transfected with genes encoding immunostimulatory cytokines (such as, but not limited to, GM-CSF)).

[0127] Dosage and administration schedule

[0128] The anti-PD-1 antibodies of the present invention can be administered by continuous infusion or by intermittent doses. The single administration dose range may be about 0.01 to about 20 mg / kg, about 0.1 to about 10 mg / kg individual body weight, or about 120 mg to about 480 mg fixed dose. For example, the dose may be about 0.1, about 0.3, about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9 or about 10 mg / kg individual body weight, or a fixed dose of about 120 mg, 240 mg, 360 mg or 480 mg. Dosage regimens are generally designed to achieve such exposure, which results in sustained receptor occupancy (RO) based on the typical pharmacokinetic properties of Ab. A representative dosing regimen may be about once a week, about once every two weeks, about once every three weeks, about once every four weeks, about once a month, or longer. In some embodiments, anti-PD-1 antibodies are administered to an individual about once every three weeks. In some embodiments, anti-PD-1 antibodies are administered to an individual about once every two weeks.

[0129] In some embodiments, the anti-PD-1 antibody of the present invention is Teplizumab, and its single administration dose is selected from about 1 to about 5 mg / kg of individual body weight. In some embodiments, the single administration dose of Teplizumab is selected from a dose of about 1 mg / kg, 2 mg / kg, 3 mg / kg, 3 mg / kg, 4 mg / kg and 5 mg / kg of body weight, or a fixed dose of 120 mg, 240 mg and 360 mg, administered intravenously. In some preferred embodiments, Teplizumab is administered as a liquid drug, and the selected dose of the drug is administered by intravenous infusion over a period of 30 to 60 minutes. In some embodiments, Teplizumab is administered at a fixed dose of about 3 mg / kg or about 240 mg once every three weeks (Q3W) by intravenous infusion over a period of 30 minutes. In some embodiments, Teplizumab is administered at a fixed dose of about 3 mg / kg or about 240 mg once every two weeks (Q3W) by intravenous infusion over a period of 30 minutes.

[0130] Each therapeutic agent of the pharmaceutical combination of the present invention can be administered simultaneously (i.e., in the same pharmaceutical composition), concurrently (i.e., in separate pharmaceutical formulations, administered one after the other in any order), or sequentially in any order. Sequential administration is particularly useful when the therapeutic agents in the pharmaceutical combination are available in different dosage forms (one drug is a tablet or capsule and the other drug is a sterile liquid formulation) and / or on different dosing schedules (e.g., a chemotherapeutic agent is administered at least daily and a biotherapeutic is administered less frequently (e.g., once a week, once every two weeks, or once every three weeks)).

[0131] In some embodiments, at least one of the therapeutic agents in the drug combination is administered using the same dosing regimen (therapeutic dose, frequency, and duration) that is typically used when the agents are used as monotherapy to treat the same tumor. In other embodiments, the patient receives a lesser total amount of at least one therapeutic agent in the combination therapy, e.g., a smaller dose, less frequent dose, and / or a shorter treatment duration, than when the agents are used as monotherapy.

[0132] Each therapeutic agent in the pharmaceutical combination of the present invention can be administered orally or parenterally, which includes intravenous, intramuscular, intraperitoneal, subcutaneous, rectal, topical and transdermal routes of administration.

[0133] The gemcitabine of the present invention is administered at its approved or recommended dose, and treatment is continued until clinical effect is observed or until unacceptable toxicity or disease progression occurs. In some embodiments, the single administration dose of gemcitabine of the present invention is selected from about 600 mg to about 1400 mg / m 2 In some embodiments, a single dose of gemcitabine is selected from about 800 mg / m 2 , 900mg / m 2 , 1000mg / m 2 , 1100mg / m 2 and 1200 mg / m 2 Any dose in body surface area. Representative dosing schedules may be about once every week, once every two weeks, once every three weeks, twice every three weeks, once every four weeks, or once a month. In some embodiments, gemcitabine is administered to an individual twice every three weeks. In some embodiments, gemcitabine is administered on day 1 and day 8 of each treatment cycle. In some embodiments, gemcitabine is administered at about 1000 mg / m 2 body surface area, twice every three weeks.

[0134] Cisplatin of the present invention is administered at its approved or recommended dose, and treatment is continued until the disease maintenance phase is reached, or until unacceptable toxicity or disease progression occurs. In some embodiments, a single dose of cisplatin of the present invention is selected from about 40 mg to about 120 mg / m 2In some embodiments, a single dose of cisplatin is selected from about 60 mg / m 2 , 70mg / m 2 , 80mg / m 2 , 90mg / m 2 and 100 mg / m 2 Any dose over a given body surface area. Representative dosing regimens may be about once a week, once every two weeks, once every three weeks, once every four weeks, or once a month. In some embodiments, cisplatin is administered to an individual once every three weeks. In some embodiments, cisplatin is administered at about 80 mg / m 2 body surface area, administered once every three weeks.

[0135] In some embodiments, toripalimab is administered at a fixed dose of about 240 mg Q3W and gemcitabine is administered at a fixed dose of about 1000 mg / m 2 body surface area, twice every three weeks, cisplatin is administered at a dose of approximately 80 mg / m 2 Body surface area, administered Q3W.

[0136] In some embodiments, on the day of toripalimab administration, gemcitabine may be administered before or after toripalimab administration, and cisplatin may be administered before or after toripalimab administration.

[0137] The administration cycles of the anti-PD-1 antibody, gemcitabine, and cisplatin of the present invention may be the same or different, and may be one week, two weeks, three weeks, one month, two months, three months, four months, five months, six months, or longer. Optionally, the duration of each administration cycle may be the same or different, and the intervals between each administration cycle may be the same or different. For example, in some embodiments, toripalimab is administered at a fixed dose of about 240 mg once every three weeks, and gemcitabine is administered at a fixed dose of about 1000 mg / m 2 body surface area, twice every three weeks, cisplatin is administered at a dose of approximately 80 mg / m 2 Body surface area, administered once every three weeks, and the dosing cycle for all three is three weeks.

[0138] Treatment methods and uses

[0139] The present invention provides use of the aforementioned anti-PD-1 antibody or antigen-binding fragment thereof of the present invention and optionally gemcitabine and cisplatin in the preparation of a medicament for preventing or treating malignant tumors.

[0140] The present invention provides a method for preventing or treating malignant tumors, comprising administering to an individual in need thereof an effective amount of an anti-PD-1 antibody or antigen-binding fragment thereof of the present invention, optionally with gemcitabine and cisplatin. The effective amount includes a prophylactic effective amount and a therapeutically effective amount. In a preferred embodiment, the dosing regimen (including dosage, frequency of administration, and order of administration, etc.) of the preventive or therapeutic method is as described in any of the embodiments above.

[0141] The present invention provides the aforementioned anti-PD-1 antibody or antigen-binding fragment thereof of the present invention, for use in preventing or treating malignant tumors. The present invention also provides a pharmaceutical combination of the aforementioned anti-PD-1 antibody or antigen-binding fragment thereof of the present invention, gemcitabine, and cisplatin, for use in preventing or treating malignant tumors.

[0142] The malignant tumor described in the present invention may be as described in any of the aforementioned embodiments; preferably, the malignant tumor described in the present invention is nasopharyngeal carcinoma, preferably, the malignant tumor described in the present invention is recurrent or metastatic nasopharyngeal carcinoma.

[0143] Preferably, the method, use, anti-PD-1 antibody and drug combination according to any embodiment of the present invention are particularly suitable for keratinizing nasopharyngeal carcinoma and non-keratinizing nasopharyngeal carcinoma, preferably keratinizing nasopharyngeal carcinoma.

[0144] Preferably, the methods, uses, anti-PD-1 antibodies, and drug combinations described in any embodiment of the present invention are particularly suitable for malignant tumors with positive PD-L1 expression in immunohistochemical staining analysis of tumor tissue sections; preferably, malignant tumors with PD-L1>25% in immunohistochemical staining analysis of tumor tissue sections.

[0145] Preferably, the methods, uses, anti-PD-1 antibodies and drug combinations described in any embodiment of the present invention are particularly suitable for malignant tumors in which CCND1, FGF14, FGF3 or FGF4 chromosome 11q13 region gene amplification is not detected in peripheral blood circulating tumor DNA or tumor tissue.

[0146] Preferably, the method, use, anti-PD-1 antibody and drug combination described in any embodiment of the present invention are particularly suitable for malignant tumors in which the number of DNA copies of EBV in peripheral blood (on the 28th day of treatment) is reduced by more than two times.

[0147] Preferred anti-PD-1 antibodies for malignant tumors may be as described in any embodiment herein, more preferably an antibody whose light chain CDRs are the amino acids shown in SEQ ID NOs: 1, 2, and 3, and whose heavy chain CDRs are the amino acids shown in SEQ ID NOs: 4, 5, and 6, more preferably a monoclonal antibody comprising a light chain variable region shown in SEQ ID NO: 7 and a heavy chain variable region shown in SEQ ID NO: 8, more preferably a monoclonal antibody comprising a light chain shown in SEQ ID NO: 9 and a heavy chain shown in SEQ ID NO: 10, more preferably humanized antibodies 38, 39, 41, and 48 described in WO2014206107, and most preferably toripalimab.

[0148] In particularly preferred embodiments, the present invention provides a method for preventing or treating nasopharyngeal carcinoma, comprising administering a therapeutically effective amount of toripalimab or the drug combination described herein to a nasopharyngeal carcinoma patient; preferably, the patient is PD-L1 positive. In certain embodiments, the preferred nasopharyngeal carcinoma is keratinizing nasopharyngeal carcinoma; in certain embodiments, the preferred nasopharyngeal carcinoma patient is one in which no amplification of CCND1, FGF14, FGF3, or FGF4 genes in the chromosome 11q13 region is detected in circulating tumor DNA or tumor tissue; in certain embodiments, the preferred patient is one in which the number of EBV DNA copies in peripheral blood decreases by more than two-fold after 28 days of treatment.

[0149] In particularly preferred embodiments, the present invention provides the use of an anti-PD-1 antibody or antigen-binding fragment thereof, or the drug combination described herein, in the preparation of a medicament for the prevention or treatment of nasopharyngeal carcinoma. Preferably, the nasopharyngeal carcinoma tumor tissue section is positive for PD-L1 expression in immunohistochemical staining analysis. In certain embodiments, the preferred nasopharyngeal carcinoma is one in which no amplification of the CCND1, FGF14, FGF3, or FGF4 chromosome 11q13 region gene is detected in circulating tumor DNA or tumor tissue in peripheral blood. In certain embodiments, the preferred nasopharyngeal carcinoma is one in which the number of EBV DNA copies in peripheral blood decreases by more than two-fold after 28 days of treatment.

[0150] The therapeutic agents described in the present invention may constitute pharmaceutical compositions, such as pharmaceutical compositions containing the anti-PD-1 antibodies described herein and / or other anticancer agents other than the anti-PD-1 antibodies, and other pharmaceutically acceptable carriers. As described herein, "pharmaceutically acceptable carriers" include any and all physiologically compatible solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, etc. Preferably, carriers suitable for compositions containing anti-PD-1 antibodies are suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal or epidermal administration, such as by injection or infusion, while carriers for compositions containing other anticancer agents are suitable for parenteral administration, such as oral administration. The pharmaceutical compositions of the present invention may contain one or more pharmaceutically acceptable salts, antioxidants, water, non-aqueous carriers, and / or adjuvants such as preservatives, wetting agents, emulsifiers and dispersants. In a preferred embodiment, the other anticancer agents other than the anti-PD-1 antibodies include gemcitabine and cisplatin.

[0151] The dosage regimen is adjusted to provide the best desired response, such as maximum therapeutic response and / or minimum adverse effect. For anti-PD-1 antibodies, including administration in combination with another anticancer agent, the dosage range may be about 0.01 to about 20 mg / kg, about 0.1 to about 10 mg / kg of individual body weight, or a fixed dose of 120 mg, 240 mg, 360 mg, or 480 mg. For example, the dosage may be about 0.1, about 0.3, about 1, about 2, about 3, about 5, or about 10 mg / kg of individual body weight. Dosage regimens are typically designed to achieve such exposure that results in sustained receptor occupancy (RO) based on the typical pharmacokinetic properties of Ab. A representative dosage regimen may be about once a week, about once every two weeks, about once every three weeks, about once every four weeks, about once a month, or longer. In some embodiments, anti-PD-1 antibodies are administered to an individual about once every two weeks.

[0152] Predicting the efficacy of anti-PD-1 antibodies in treating malignant tumors

[0153] As used herein, the term "gene amplification" refers to a process in which the copy number of a gene encoding a specific protein is selectively increased while the number of copies of other genes is not proportionally increased. Under natural conditions, gene amplification occurs by excising repetitive sequences of a gene from chromosomes and replicating them extrachromosomally in plasmids, or by generating RNA transcripts from all repetitive sequences of ribosomal RNA, which are then transcribed to generate additional copies of the original DNA molecule. Some embodiments of the present invention disclose gene sequencing analysis.

[0154] In some embodiments of the present invention, the subjects described herein have certain unique gene amplifications, such as amplification of CCND1, FGF14, FGF3, or FGF4 genes in the chromosome 11q13 region. In some embodiments of the present invention, amplification of CCND1, FGF14, FGF3, or FGF4 genes in the chromosome 11q13 region indicates that the patient will not respond well to treatment with the anti-PD-1 antibody described herein alone.

[0155] In some embodiments of the present invention, the number of EBV DNA copies in the peripheral blood of some subjects decreased on treatment day 28. In some embodiments of the present invention, a decrease of more than two-fold in the number of EBV DNA copies in the peripheral blood indicates that the patient will have a better therapeutic effect when treated with the anti-PD-1 antibody of the present invention.

[0156] Therefore, the present invention provides a method for predicting the effect of using the anti-PD-1 antibody of the present invention, especially toripalimab, on treating malignant tumors in an individual, comprising detecting biomarkers in the patient's peripheral blood before treatment, wherein the biomarkers are selected from but not limited to CCND1, FGF14, FGF3 or FGF4 chromosome 11q13 region mutations, or detecting the number of EBV DNA copies in the patient's peripheral blood on the 28th day of treatment.

[0157] The present invention also includes a method for predicting the efficacy of anti-PD-1 antibody treatment in a tumor patient by detecting the presence of CCND1, FGF14, FGF3, or FGF4 gene amplification in the chromosome 11q13 region. Preferably, the presence of CCND1, FGF14, FGF3, or FGF4 gene amplification in the chromosome 11q13 region indicates that the tumor patient is not suitable for treatment with an anti-PD-1 antibody alone. Preferably, the tumor patient is selected from patients with nasopharyngeal carcinoma.

[0158] The present invention also includes a method for predicting the efficacy of anti-PD-1 antibody treatment in a tumor patient by measuring the number of EBV DNA copies in the patient's peripheral blood on day 28 of treatment. Preferably, a decrease of more than twofold in the number of EBV DNA copies in the peripheral blood (number of EBV DNA copies in the peripheral blood before treatment on day 0 / number of EBV DNA copies in the peripheral blood on day 28 ≥ 2) indicates that the tumor patient is suitable for treatment with an anti-PD-1 antibody. Preferably, the tumor patient is selected from patients with nasopharyngeal carcinoma.

[0159] The present invention also includes the use of reagents for detecting mutations in the CCND1, FGF14, FGF3, or FGF4 chromosome 11q13 region in the preparation of a kit for predicting the efficacy of anti-PD-1 antibodies in treating malignant tumors. Such reagents include, but are not limited to, reagents routinely used in testing, including, but not limited to, primers, probes, and reagents required for PCR.

[0160] The present invention also includes the use of reagents for detecting the number of EBV DNA copies in peripheral blood in the preparation of a kit for predicting the efficacy of anti-PD-1 antibodies in treating malignant tumors. Such reagents include, but are not limited to, reagents routinely used in testing, including, but not limited to, primers, probes, and reagents required for PCR.

[0161] medicine box

[0162] The present invention also provides a drug kit containing one or more single drug dosage units of an anti-PD-1 antibody or antigen-binding fragment thereof as described in any embodiment herein; or containing one or more single drug dosage units of an anti-PD-1 antibody or antigen-binding fragment thereof as described in any embodiment herein, one or more single drug dosage units of gemcitabine as described in any embodiment herein, and one or more single drug dosage units of cisplatin as described in any embodiment herein.

[0163] In some embodiments, the kit contains one or more single-dose units of the drug combination described in any embodiment herein. In some embodiments, the kit contains one or more groups of drug preparations, each group of drug preparations is a 3-week dosing dose, including 1 dose of toripalimab, 2 doses of gemcitabine and 1 dose of cisplatin; preferably, the toripalimab monoclonal antibody is a fixed dose of about 240 mg, and the 2 doses of gemcitabine are sufficient to administer at a dose of about 1000 mg / m 2 The 1 dose of cisplatin is sufficient to deliver approximately 80 mg / m2 of cisplatin to the body surface area twice. 2 Apply once per body surface area.

[0164] The kit of the present invention can be used in the treatment of malignant tumors, such as those described in any embodiment herein, and in particular, nasopharyngeal carcinoma. Preferably, the amount of the anticancer active ingredient contained in the kit of the present invention is sufficient for one or more courses of treatment (e.g., 2-8 courses) according to the treatment methods described herein. In the case of the drug combination of the present invention, the course of treatment includes at least one administration of toripalimab, two administrations of gemcitabine, and one administration of cisplatin.

[0165] Abbreviations

[0166] Throughout the description and examples of the present invention, the following abbreviations are used:

[0167] BID one dose, twice daily

[0168] CDR complementarity determining region

[0169] DFS disease-free survival

[0170] FR framework region

[0171] IgG immunoglobulin G

[0172] IHC immunohistochemistry

[0173] OR overall response rate

[0174] ORR objective response rate

[0175] OS overall survival rate

[0176] PD disease progression

[0177] PFS progression-free survival

[0178] PR partial response

[0179] CR (complete remission)

[0180] SD stable disease

[0181] DLT dose-limiting toxicity

[0182] MTD maximum tolerated dose

[0183] AE adverse event

[0184] Q2W One dose every two weeks

[0185] QD one dose per day

[0186] CSD long-term sunshine type

[0187] non-CSD non-long-term sunshine type

[0188] IRC Independent Review Committee

[0189] TRAEs Treatment-related adverse events

[0190] SAE serious adverse event

[0191] RO receptor occupancy

[0192] UC urothelial carcinoma

[0193] RCC Renal Cell Carcinoma

[0194] MM metastatic melanoma

[0195] RECIST criteria for evaluating the efficacy of solid tumors

[0196] irRECIST immune-related solid tumor response evaluation criteria

[0197] DOR duration of response

[0198] MSI microsatellite instability

[0199] BICR: Double-blind independent central review

[0200] The present invention is further illustrated by the following examples, which should not be construed as limiting the present invention.The contents of all references cited throughout this application are expressly incorporated herein by reference.

[0201] Example

[0202] Example 1: Clinical study of anti-PD-1 antibody treatment of nasopharyngeal carcinoma

[0203] Eligible subjects must be (1) aged 18 years or older, (2) have recurrent or metastatic nasopharyngeal carcinoma, (3) be refractory to standard systemic therapy or have progressed after 6 months of chemoradiotherapy, (4) have an ECOG performance status of 0 or 1, (5) have normal organ function within 10 days of treatment initiation, (6) have no history of autoimmune diseases or other malignancies, and (7) have not received any previous anti-PD-1 / or anti-PD-L1 immunotherapy.

[0204] Subjects must have evaluable lesions according to RECIST v 1.1 criteria, have not received anti-tumor monoclonal antibody treatment within 4 weeks before treatment, have not received any anti-tumor drug treatment within 2 weeks before treatment, and have not received systemic steroid treatment within 7 days before the start of treatment.

[0205] From December 22, 2016, to February 19, 2019, a total of 279 patients with recurrent or metastatic NPC were screened at 17 centers in mainland China, and 190 patients were enrolled in this study. The mean age was 46.4 years, and the majority of patients were male (n = 158, 83.2%). Of the two histological subtypes, 182 (95.8%) were non-keratinizing NPC and 8 (4.2%) were keratinizing NPC. 116 patients (61.1%) had received at least two prior systemic therapies. The demographic data of the enrolled subjects are shown in Table 1.

[0206] Table 1: Demographic data of enrolled subjects

[0207]

[0208] Note: *PD-L1 positivity is defined as PD-L1 expression of ≥1% of tumor cells using SP142 IHC staining.

[0209] Test drug: anti-PD-1 antibody toripalimab (WO2014206107).

[0210] Enrolled subjects received toripalimab 3 mg / kg intravenously every two weeks (Q2W) until confirmed disease progression, intolerable toxicity, withdrawal of individual consent, investigator decision to discontinue treatment, or until the end of 24 months of treatment.

[0211] Patients were assessed every 8 weeks during the first year and every 12 weeks thereafter according to RECIST v1.1 and immune-related response evaluation criteria in solid tumors (irRECIST). Patients were assessed every 3 months after treatment discontinuation.

[0212] Clinical design:

[0213] This is a single-arm, Phase II, open-label clinical trial to evaluate the safety and anti-tumor activity of anti-PD-1 antibodies in the treatment of patients with recurrent or metastatic nasopharyngeal carcinoma.

[0214] 1.1 Safety Study:

[0215] As of February 19, 2020, 12 months after the last patient was admitted, the median number of doses of toripalimab received by patients was 8 (range: 1 to 69). 181 (95.3%) patients experienced treatment-emergent adverse events (TEAEs), of which 141 (74.2%) experienced treatment-related adverse events (TRAEs). Common (>5%) TRAEs are shown in Table 2. 63 (33.2%) patients experienced TEAEs of grade 3 or higher, while 27 (14.2%) patients experienced treatment-related TRAEs of grade 3 or higher. 4 (2.1%) patients discontinued due to TRAEs, and 7 (3.7%) patients had dose interruptions due to TRAEs. Immune-related adverse reactions (AEs) included 45 patients (23.7%) with hypothyroidism, 5 patients (2.6%) with hyperthyroidism, 3 patients (1.6%) with abnormal liver function, 3 patients (1.6%) with interstitial lung disease, 1 patient (0.5%) with dermatomyositis, and 1 patient (0.5%) with autoimmune myocarditis.

[0216] Table 2: Common (>5%) adverse reactions associated with toripalimab treatment (n=190)

[0217] N (%) All Grade 1 Grade 2 Grade 3 Grade 4 Grade 5 All adverse reactions 141 (74.2) 55 (28.9) 59 (31.1) 17 (8.9) 4 (2.1) 6 (3.2) Hypothyroidism 45 (23.7) 19 (10.0) 26 (13.7) 000 Anemia 29 (15.3) 15 (7.9) 12 (6.3) 2 (1.1) 000 AST increased 29 (15.3) 26 (13.7) 3 (1.6) 000 ALT increased 26 (13.7) 21 (11.1) 5 (2.6) 000 Fatigue 25 (13.2) 18 (9.5) 5 (2.6) 2 (1.1) 00 Proteinuria 24 (12.6) 24 (12.6) 0000 Leukopenia 19 (10.0) 8 (4.2) 11 (5.8) 000 Fever 18 (9.5) 13 (6.8) 5 (2.6) 000 Pruritus 16 (8.4) 14 (7.4) 2 (1.1) 000 Rash 12 (6.3) 8 (4.2) 4 (2.1) 000 Neutropenia 10 (5.3) 6 (3.2) 3 (1.6) 1 (0.5) 00

[0218] 1.2 Anti-tumor activity studies:

[0219] As of February 19, 2020, of the 190 patients enrolled, 94 (49.5%) had died, 78 (41.1%) had discontinued treatment, and 18 (9.5%) remained on study. The median duration of treatment was 3.7 months (range, 0.2 to 34.8 months). Among the 190 patients, the objective response rate (ORR) was 20.5% (95% CI, 15.0-27.0) according to IRCIST / RECIST v1.1, including 5 complete responses (CRs), 34 partial responses (PRs), and 37 sudden changes in disease, for a disease control rate (DCR) of 40.0% (95% CI, 33.0-47.3). By IRCIST / irRECIST assessment, the ORR was 20.5% (95% CI, 15.0-27.0), and the DCR was 47.9% (95% CI, 40.6-55.2) (Table 3).

[0220] Table 3: Clinical efficacy assessed according to RECIST v1.1 or irRECIST criteria

[0221]

[0222] Note:

[0223] *ORR = (CR + PR) / total × 100%;

[0224] **DCR = (CR + PR + SD) / total × 100%;

[0225] CR: complete response; PR: partial response; SD: stable disease; PD: progressive disease; NE: not evaluable; ORR: objective response rate; DCR: disease control rate; CI: confidence interval.

[0226] Target lesions decreased relative to baseline in 73 subjects (38.4%), of whom 48 subjects (25.3%) achieved a greater than 30% reduction in target lesions relative to baseline ( Figures 1 , 1a and 1b ). The median duration of response was 1.8 months (95% CI: 1.8-2.1). The median progression-free survival (mPFS) was 1.9 months (95% CI: 1.8-3.5) ( Figure 2 , 2a ). The median overall survival (mOS) was 17.4 months (95% CI: 11.7-22.9) ( Figure 2 , 2b ). Responses were durable, with a median duration of response ( DOR) of 12.8 months (95% CI: 9.4-NE) ( Figure 2 , 2c ). For patients who had received at least two prior therapies (n=116), the ORR was 21.6% (95% CI 14.5-30.1), the median DOR was 21.5 months, the mPFS was 2.0 months, and the mOS was 15.1 months.

[0227] Since only 7 PR / CR subjects and 11 SD subjects died, the median OS for subjects who experienced an objective response (n=39) or stable disease (n=38) was not reached. The median OS for subjects with progressive disease (n=113) was 8.4 months.

[0228] 1.3 Immunogenicity

[0229] Antidrug antibody (ADA) testing was performed in 190 patients. Seven patients (3.7%) were ADA-positive, including four consecutive positive samples. There were no significant differences in the incidence of AEs, SAEs, grade 3 or higher AEs, discontinuations or dose delays, or clinical efficacy between ADA-positive and ADA-negative patients.

[0230] 1.4 Histological subtypes

[0231] Based on IRC assessment, the ORR for keratinizing NPC (n=8) was significantly superior to that for non-keratinizing NPC (n=182), at 62.5% and 18.7%, respectively (p=0.01). PFS was also significantly superior in keratinizing NPC compared with non-keratinizing NPC, at 16.6 months and 1.9 months, respectively (HR=0.46 (95% CI: 0.25-0.85), p=0.013). OS was not reached and 15.1 months, respectively, with no statistically significant difference (HR=0.51 (95% CI: 0.21-1.24), p=0.14) (Figures 2d and 2e).

[0232] Example 2: Study on the correlation between biomarkers and clinical efficacy

[0233] 2.1 PD-L1 expression in tumors

[0234] The PD-L1 expression status in tumor biopsies was determined by SP142 IHC staining, and the tumor proportion score (TPS) was >1%, which was determined to be positive. Among the 190 patients, 48 ​​(25.3%) were PD-L1 positive and 134 (70.5%) were PD-L1 negative (Figure 3, 3a). The PD-L1 expression status of 8 patients (4.2%) was unknown. Among the PD-L1+ patients, 21 (11.1%) were determined to have high PD-L1 expression (>25%). According to the histological subtype, the PD-L1+ percentage was significantly higher in keratinizing NPC (75.0%) than in non-keratinizing NPC (24.1%), p=0.0047. The ORR values ​​of PD-L1+ patients were higher than those of PD-L1- patients, with ORRs of 27.1% and 19.4%, respectively, but the difference was not statistically significant (p=0.31). In patients with PD-L1 > 25%, the difference in ORR was more pronounced (38.1% vs. 19.3%, p value = 0.08) (Figure 3, 3a). Patients with PD-L1 > 25% also had better PFS and OS than those with PD-L1 ≤ 25%, with mPFS of 7.2 months and 1.9 months, respectively, HR = 0.64 (95% CI: 0.40-1.02), p = 0.059; mOS was not reached and 15.1 months, respectively, HR = 0.57 (95% CI: 0.31-1.05), p = 0.071 (Figure 3, 3b and 3c), but the differences were not statistically significant.

[0235] 2.2 Tumor mutation burden (TMB) analysis

[0236] Whole exome sequencing (WES) was performed on tumor biopsy tissue and paired peripheral blood samples from the subjects to identify tumor-specific mutations. Valid WES results were available for 174 patients. In this NPC patient population, TMB was very low, with a median TMB of 0.95 mutations / million base pairs (Muts / Mb). Only one MSI-high patient and four patients had a TMB exceeding 10 mutations / Mb, while the remaining patients had a TMB not exceeding 5.8 mutations / Mb. This study selected a cutoff of the top 10% or 20% of TMB values ​​(2.9 and 2.0 Muts / Mb, respectively) for the evaluation of clinical response. The ORRs of the top 10% and top 20% of patients were 17.6% and 14.3%, respectively (Figure 3, 3a). Four patients with a TMB exceeding 10 Muts / Mb, including one MSI-high patient, had the best response with progressive disease. Furthermore, at 1.9 months, the 10% of patients with the highest TMB values ​​had similar PFS to the 90% of patients with the lowest TMB values ​​(Figure 3, 3d). Conversely, patients with high TMB had lower OS values ​​than those with low TMB, at 9.2 months and 17.4 months, respectively, but the difference was not statistically significant (Figure 3, 3e). In summary, in this study, TMB was not associated with clinical response in patients with advanced NPC who received toripalimab monotherapy.

[0237] 2.3 Genomic mutation analysis

[0238] The most frequently altered genes (≥10%) identified by WES included CDKN2A (20%), TP53 (13%), NFKB1A (13%), CDKN2B (11%), ETV6 (11%), and MCL1 (10%) (Figure 4). The correlation between genomic alterations and clinical efficacy was analyzed. The study found that 11 patients with genomic amplification of the chromosome 11q13 region of CCND1 (n=11) and / or FGF14, FGF3, or FGF4 had an ORR of 0% with toripalimab. Nineteen patients had mutations in ETV6, and the ORR was only 5.3%.

[0239] 2.4 Plasma EBV DNA copy number

[0240] Plasma was collected from patients before treatment, and EBV DNA copy number was analyzed every 4 weeks by qRT-PCR. Patients with a baseline EBV titer <10,000 IU / mL had a higher ORR than patients with an EBV titer ≥10,000 IU / mL, 26.7% and 15.4%, respectively, p = 0.088. Dynamic plasma EBV DNA copy number during treatment was collected from 149 patients (Figure 5). The study found that in patients with objective response (n = 34), the median decrease in plasma EBV DNA copy number from baseline to the lowest copy number was 31 times, while in patients with stable disease (n = 35), it decreased by 3 times, and there was no change in patients with progressive disease (n = 80) (Figure 5). In addition, patients whose plasma EBV DNA copy number decreased by more than 2-fold on day 28, two weeks before the first radiographic assessment of clinical activity (n = 60), had a significantly better clinical response rate than those whose plasma EBV DNA copy number decreased by less than 2-fold (n = 88), with an ORR of 48.3% and 5.7%, respectively (p = 0.0001). In contrast, the 14 patients who experienced disease progression had at least a 2-fold increase in plasma EBV DNA copy number at least 3 months before radiographic confirmation of disease progression (Figure 5).

[0241] Example 3: Clinical study of anti-PD-1 antibody combined with gemcitabine-cisplatin (GP) in the treatment of nasopharyngeal carcinoma

[0242] Subjects

[0243] Eligible subjects must meet the following criteria: age 18-75 years; have recurrent or metastatic NPC (stage IVB according to the eighth edition of the International Union Against Cancer and American Joint Committee on Cancer staging system); have not received prior systemic chemotherapy for recurrent or metastatic disease; have at least one measurable lesion according to RECIST version 1.1; have a life expectancy of more than 3 months; have an Eastern Cooperative Oncology Group performance status score of 0 or 1; and have normal organ function. For NPC that relapsed after systemic treatment, the interval between recurrence and the last dose of the previous radiation or chemotherapy must have been more than 6 months according to the National Cancer Institute Common Terminology Criteria (CTCAE version 5.0), and any toxicity of the prior treatment must have been reduced to grade 0 or 1.

[0244] Exclusion criteria included: a history of severe hypersensitivity reaction to any monoclonal antibody, gemcitabine, cisplatin, or any component of toripalimab; active or untreated central nervous system metastases or spinal cord compression; potential risk of major bleeding due to necrotic lesions; uncontrolled pleural or pericardial effusion; uncontrolled ascites; uncontrolled tumor-related pain; uncontrolled or symptomatic hypercalcemia; prior malignancy other than NPC within the past 5 years, excluding those with minimal risk of metastasis or death after prospective treatment; prior treatment with monoclonal antibodies targeting PD-1 / PD-L1 / CTLA4; and use of anti-tumor traditional herbal medicines within the past 4 weeks. ; history of major surgery in the past 28 days or anticipated major surgery during the study; history of autoimmune disease; use of systemic immunostimulants within 4 weeks prior to treatment or within the half-life of the drug; use of systemic corticosteroids or immunosuppressive drugs within 2 weeks prior to treatment; history of bone marrow or solid organ transplantation; history of non-infectious pneumonitis or current pneumonitis; use of any live vaccine within the previous 4 weeks; active tuberculosis, active hepatitis B virus or hepatitis C virus infection; active human immunodeficiency virus; active major neuropathy or psychiatric disease; grade 2 or higher peripheral neuropathy; pregnant or lactating women or clinically important cardiovascular disease.

[0245] Test drug

[0246] Anti-PD-1 antibody: toripalimab, Suzhou Junmeng Biopharmaceutical Technology Co., Ltd.

[0247] Gemcitabine (Gem), Cisplatin (Cis)

[0248] Clinical design

[0249] This was a randomized, double-blind, placebo-controlled study. Randomization was performed using the Interactive VoiceWeb Remission System (IVRS). Patients were stratified according to Eastern Cooperative Oncology Group performance status (0 or 1) and disease stage (local recurrence versus primary metastasis) before enrollment.

[0250] The subjects were randomly divided into group A and group B in a 1:1 ratio. Group A received toripalimab combined with gemcitabine (Gem) and cisplatin (Cis) once every 3 weeks (Q3W), and group B received placebo combined with gemcitabine and cisplatin once every 3 weeks (Q3W). All drugs were given by intravenous infusion. Patients will receive toripalimab (240 mg) or placebo on day 1 of each 3-week cycle, and gemcitabine (1000 mg / m2) on days 1 and 8. 2 body surface area), and received cisplatin (80 mg / m 2Body surface area). Chemotherapy will continue until disease progression, intolerable toxicity, noncompliance, withdrawal of consent, or a maximum of 6 cycles, whichever occurs first during the chemotherapy phase. During the maintenance phase, patients will receive toripalimab (240 mg) (Arm A) or placebo (Arm B) every 3 weeks as maintenance therapy until disease progression, intolerable toxicity, withdrawal of consent, or the investigator's judgment, or a maximum of 2 years. Crossover studies are not permitted as part of the study.

[0251] end

[0252] The primary endpoint was progression-free survival (PFS) in the intention-to-treat population, defined as the time from randomization to the first documented disease progression due to any cause or death, whichever occurred first. Secondary endpoints included overall survival (OS) in the ITT population; objective response rate (ORR), defined as the proportion of patients with confirmed complete response or partial response; duration of response (DoR), defined as the time between the first documented response and the first evidence of progressive disease; disease control rate (DCR), defined as the proportion of patients with a best response of complete response (CR), partial response (PR), or stable disease (SD); and 1- and 2-year PFS and OS rates.

[0253] Evaluate

[0254] Baseline tumor assessments included CT scans of the nasopharynx, neck, chest, and abdomen (with oral / intravenous contrast unless contraindicated) or MRI or whole-body positron emission tomography (PET) / CT scans. A bone scan was performed if clinically indicated. All known sites of disease were recorded at screening and reassessed at each subsequent tumor assessment. The same radiographic procedures used to assess baseline disease sites were used throughout the study. Tumor assessments were performed every 6 weeks for the first 12 months and then every 9 weeks until disease progression, loss of clinical benefit, withdrawal of consent, initiation of new anticancer therapy, death, or investigator discontinuation of the study, whichever occurred first.

[0255] Clinical response was assessed by investigators and blinded independent central review (BICR) according to RECIST v1.1 and irRECIST.

[0256] 3.1 Treatment of Subjects

[0257] Between November 10, 2018, and October 20, 2019, 408 subjects were screened from 48 sites in China and Singapore (Figure 6). A total of 289 eligible subjects were randomly assigned to the toripalimab combination group (Group A, n=146) or the placebo combination group (Group B, n=143). Although the proportion of smokers and drinkers in the toripalimab group was higher than that in the placebo group, the baseline demographics and disease characteristics of the two groups were generally balanced (52.1% vs. 41.3%, P=0.077 and 20.5% vs. 12.6%, P=0.082). However, the difference was not statistically significant. Based on the definition of tumor cell or immune cell positivity rate ≥1%, 74.7% of the subjects from the toripalimab group and 76.2% of the subjects in the placebo group stained positive for PD-L1 expression. The demographic data of the enrolled subjects are shown in Table 4.

[0258] During the chemotherapy phase, all patients received at least one dose of study drug. By the cutoff date, 56 (19.4%) participants had discontinued study treatment (31 in the toripalimab group and 25 in the placebo group). Both groups received six cycles of chemotherapy. After completion of chemotherapy, 231 (79.9%) patients continued to receive maintenance therapy (113 in the toripalimab group and 118 in the placebo group). Participants received an average of 12 cycles of toripalimab and 11 cycles of placebo.

[0259] Table 4: Demographic data of enrolled subjects

[0260]

[0261] 3.2 Progression-free survival

[0262] In a pre-specified interim analysis of 128 patients with disease progression or death, as assessed by BICR according to RECIST v1.1, the median treatment duration was 39 weeks in the toripalimab and 36 weeks in the placebo groups. The median PFS was 11.7 months (95% CI, 11.0 to NE) in the toripalimab group and 8.0 months (95% CI, 7.0 to 9.5) in the placebo group. PFS was significantly improved in the toripalimab group compared with placebo (hazard ratio for progression or death, 0.52; 95% CI, 0.36 to 0.74; two-sided P = 0.0003) (Figure 6, 6a). The estimated 1-year PFS was 49.4% (95% CI, 36.4 to 61.1) in the toripalimab group and 27.9% (95% CI, 18.0, 38.8) in the placebo group, a difference of 21.4% (95% CI, 5.1 to 37.8). Across all relevant subgroups, including all PD-L1 subgroups, superior PFS with toripalimab to placebo was observed (Figure 6, 6c). For patients with PD-L1-positive tumor cells (TC) ≥ 1% or immune cells (IC) ≥ 1%, the hazard ratio for progression or death between the toripalimab group and the placebo group was 0.59 (95% CI, 0.39 to 0.89), and the median PFS was 11.4 vs. 8.2 months. For patients with <1% PD-L1-positive tumor cells (TC) and <1% immune cells (IC), the hazard ratio for progression or death was 0.35 (95% CI, 0.15 to 0.81), and the median PFS was 11.0 vs. 6.0 months.

[0263] According to investigator assessment based on RECIST v1.1, treatment with toripalimab plus chemotherapy reduced the risk of death or death by 59% (HR = 0.41; 95% CI, 0.28 to 0.59; P < 0.0001) compared with placebo plus chemotherapy (Figure 6, 6b). The 1-year PFS rate was 59.5% in the toripalimab group and 20.0% in the placebo group, a difference of 39.5% (95% CI, 25.6 to 53.5).

[0264] 3.3 Overall Survival

[0265] At the interim analysis cutoff date of May 30, 2020, 29 deaths were reported: 12 (8.2%) in the toripalimab group and 17 (11.9%) in the placebo group. Median survival had not been reached in either group. The stratified hazard ratio for OS was 0.78 (95% CI, 0.37 to 1.64; P = 0.50). According to the survival update on February 18, 2021, a total of 64 deaths were reported, including 25 deaths in the toripalimab group and 39 deaths in the placebo group. The stratified hazard ratio for OS was 0.60 (95% CI, 0.36 to 1.00; P = 0.0462), and the risk of immediate death in patients in the toripalimab group was 40% lower than that in the placebo group (Figure 7). The estimated proportion of patients alive at 2 years was 77.8% (95% CI, 68.0 to 85.0) in the toripalimab group and 63.3% (95% CI, 49.8 to 74.1) in the placebo group. Median OS values ​​for either group were immature because of the limited number of OS events.

[0266] 3.4 Tumor Response

[0267] According to the BICR assessment, 28 patients (19.2%) in the toripalimab group and 16 (11.2%) in the placebo group had confirmed complete remission, and 85 (58.2%) in the toripalimab group and 79 (55.2%) in the placebo group had confirmed partial remission. The ORR in the toripalimab group was 77.4% (95% CI, 69.8 to 83.9) and in the placebo group was 66.4% (95% CI, 58.1 to 74.1) (P = 0.0335). The DCR in the toripalimab group was 87.7% (95% CI, 81.2 to 92.5) and in the placebo group was 79.7% (95% CI, 72.2, 86.0) (P = 0.0650) (Table 5). The investigator-assessed ORR was 80.8% (95% CI, 73.5 to 86.9) in the toripalimab group and 74.8% (95% CI, 66.9 to 81.7) in the placebo group.

[0268] Based on the BICR assessment in the ITT population, 114 patients in the toripalimab group and 95 in the placebo group were responders. The median duration of response was 10.0 (95% CI, 8.8 to NE) months in the toripalimab group and 5.7 (95% CI, 5.4 to 6.8) months in the placebo group (HR: 0.50; 95% CI, 0.33 to 0.78) (Table 5). As of May 30, 2020, 66% (75 / 114) of the toripalimab group and 43% (41 / 95) of the placebo group had ongoing responses (Figure 8). Based on investigator assessment of the ITT population, the median DoR was not reached (95% CI, 9.7 to NE) in the toripalimab group and was 5.8 (95% CI, 5.7 to 6.9) months in the placebo group (HR: 0.37; 95% CI, 0.24 to 0.56).

[0269] Table 5: Tumor Response to Toripalimab Versus Placebo in Combination with Gemcitabine and Cisplatin in Patients with Recurrent or Metastatic Nasopharyngeal Carcinoma According to RECIST v1.1

[0270]

[0271] 3.5 Adverse Reactions

[0272] As of May 30, 2020, the median treatment duration was 38.7 weeks in the toripalimab group and 36.0 weeks in the placebo group. The median exposure duration to cisplatin was 18.3 and 18.4 weeks in the toripalimab group and the placebo group, respectively, and the median exposure duration to gemcitabine was 19.3 and 19.6 weeks, respectively. All patients experienced at least one treatment-emergent adverse event (TEAE). The incidence of TEAEs ≥ 3 in the toripalimab group and the placebo group was 89.0% and 89.5%, respectively, and the proportion of TEAEs leading to discontinuation of toripalimab / placebo was 7.5% and 4.9%, respectively; serious adverse reactions (SAEs) (41.1% vs. 43.4%) and fatal TEAEs (2.7% vs. 2.8%) were similar between the two groups.

[0273] The most common TEAEs included leukopenia (91.1% in the toripalimab group vs 94.4% in the placebo group), anemia (88.4% vs 94.4%), neutropenia (85.6% vs 93.0%), nausea (69.2% vs 83.2%), vomiting (67.1% vs 65.7%), thrombocytopenia (63.0% vs 62.2%), and decreased appetite (53.4% ​​vs 58.7%).

[0274] The incidence of grade 3 or higher TEAEs was similar in the two groups and included leukopenia (61.6% vs 58.0%), neutropenia (57.5% vs 63.6%), anemia (47.3% vs 39.9%), thrombocytopenia (32.9% vs 28.7%), pneumonia (10.3% vs 3.5%), lymphopenia (8.9% vs 7.0%), hyponatremia (8.9% vs 4.2%), and hypokalemia (6.8% vs 7.0%).

[0275] 3.6 Conclusion

[0276] In this randomized, phase III study, we compared the efficacy and toxicity of toripalimab versus placebo in combination with GP (gemcitabine and cisplatin) chemotherapy for the treatment of recurrent or metastatic NPC. The results showed that adding toripalimab to chemotherapy resulted in a higher overall response rate and longer overall survival than chemotherapy plus placebo, with a manageable safety profile.

Claims

1. Use of an anti-PD-1 antibody or an antigen-binding fragment thereof in the preparation of a medicament for preventing or treating a malignant tumor, and use of a combination of an anti-PD-1 antibody or an antigen-binding fragment thereof with gemcitabine and cisplatin in the preparation of a medicament for preventing or treating a malignant tumor; preferably, the malignant tumor is nasopharyngeal carcinoma; more preferably, the malignant tumor is recurrent or metastatic nasopharyngeal carcinoma.

2. The use according to claim 1, It is characterized in that The malignant tumor is a malignant tumor in which PD-L1 expression is greater than 1% in immunohistochemical staining analysis of tumor tissue sections; preferably a malignant tumor in which PD-L1 expression is greater than 25% in immunohistochemical staining analysis of tumor tissue sections.

3. The use according to claim 1, It is characterized in that The malignant tumor is selected from keratinizing nasopharyngeal carcinoma and non-keratinizing nasopharyngeal carcinoma, preferably keratinizing nasopharyngeal carcinoma.

4. The use according to claim 1, It is characterized in that The nasopharyngeal carcinoma is a nasopharyngeal carcinoma in which no genomic amplification of CCND1, FGF14, FGF3 or FGF4 chromosome 11q13 region is detected in peripheral blood circulating tumor DNA or tumor tissue; or The nasopharyngeal carcinoma is a nasopharyngeal carcinoma in which the number of EBV DNA copies in peripheral blood on the 28th day of treatment decreases by more than two times compared with that before administration on the 0th day.

5. The use according to any one of claims 1 to 4, It is characterized in that The anti-PD-1 antibody or its antigen-binding fragment comprises a light chain complementary determining region with an amino acid sequence as shown in SEQ ID NOs: 1, 2 and 3, and a heavy chain complementary determining region with an amino acid sequence as shown in SEQ ID NOs: 4, 5 and 6; preferably, the anti-PD-1 antibody or its antigen-binding fragment comprises a light chain variable region with an amino acid sequence as shown in SEQ ID NO: 7, and a heavy chain variable region with an amino acid sequence as shown in SEQ ID NO: 8; more preferably, the anti-PD-1 antibody comprises a light chain with an amino acid sequence as shown in SEQ ID NO: 9, and a heavy chain with an amino acid sequence as shown in SEQ ID NO:

10.

6. The use according to any one of claims 1 to 5, It is characterized in that The anti-PD-1 antibody is selected from one or more of nivolumab, pembrolizumab, toripalimab, Sintilimab, Camrelizumab, Tislelizumab, and Cemiplimab; preferably toripalimab.

7. The use according to any one of claims 1 to 6, It is characterized in that The anti-PD-1 antibody or antigen-binding fragment thereof is administered alone, wherein the anti-PD-1 antibody or antigen-binding fragment thereof is administered at a dose of about 0.1 mg / kg to about 10.0 mg / kg of individual body weight, such as about 0.1 mg / kg, about 0.3 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 5 mg / kg or 10 mg / kg of individual body weight, or selected from a fixed dose of about 120 mg to about 480 mg, such as a fixed dose of 120 mg, 240 mg, 360 mg or 480 mg, preferably a fixed dose of 3 mg / kg of individual body weight or 240 mg; or The anti-PD-1 antibody or its antigen-binding fragment is administered in combination with gemcitabine and cisplatin, wherein the anti-PD-1 antibody or its antigen-binding fragment is administered at a dose of about 0.1 mg / kg to about 10.0 mg / kg of individual body weight, such as about 0.1 mg / kg, about 0.3 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 5 mg / kg or 10 mg / kg of individual body weight, or is selected from a fixed dose of about 120 mg to about 480 mg, such as a fixed dose of 120 mg, 240 mg, 360 mg or 480 mg, preferably 3 mg / kg of individual body weight or a fixed dose of 240 mg; the single administration dose of gemcitabine is about 600 mg / m 2 About 1400 mg / m 2 Body surface area, e.g. 800 mg / m 2 , 1000mg / m 2 or 1200 mg / m 2 body surface area; and a single administration dose of cisplatin of about 40 mg / m 2 To about 120mg / m 2 Body surface area, e.g. 60 mg / m 2 , 80mg / m 2 or 100 mg / m 2 Body surface area.

8. The use according to claim 7, It is characterized in that The anti-PD-1 antibody or antigen-binding fragment thereof is administered alone, wherein the frequency of administration of the anti-PD-1 antibody or antigen-binding fragment thereof is about once a week, once every two weeks, once every three weeks, once every four weeks or once a month, preferably once every two weeks; or The anti-PD-1 antibody or an antigen-binding fragment thereof is administered in combination with gemcitabine and cisplatin, wherein the anti-PD-1 antibody or the antigen-binding fragment thereof is administered at a frequency of about once a week, once every two weeks, once every three weeks, once every four weeks or once a month, preferably once every three weeks; and the gemcitabine is administered at a frequency of about once a week, once every two weeks, once every three weeks, twice every three weeks, once every four weeks or once a month, preferably twice every three weeks; and the cisplatin is administered at a frequency of about once a week, once every two weeks, once every three weeks, once every four weeks or once a month, preferably once every three weeks.

9. The use according to claim 8, It is characterized in that The anti-PD-1 antibody or antigen-binding fragment thereof is administered alone, wherein the dosage of the anti-PD-1 antibody or antigen-binding fragment thereof is 1 mg / kg individual body weight, 3 mg / kg individual body weight, 10 mg / kg individual body weight, or a fixed dose of 240 mg, a fixed dose of 480 mg, administered once every two weeks or three weeks; or The anti-PD-1 antibody or its antigen-binding fragment is administered in combination with gemcitabine and cisplatin, wherein the anti-PD-1 antibody or its antigen-binding fragment is administered at a fixed dose of 240 mg once every three weeks; and the single administration dose of gemcitabine is about 1000 mg / m 2 body surface area, twice every three weeks; and a single administration dose of cisplatin of about 80 mg / m 2 body surface area, once every three weeks.

10. The use according to claim 9, It is characterized in that The anti-PD-1 antibody or antigen-binding fragment thereof, gemcitabine, and cisplatin are administered in a liquid dosage form, such as an injection, via a parenteral route, such as intravenous infusion.

11. The use according to claim 10, It is characterized in that The administration cycles of the anti-PD-1 antibody or antigen-binding fragment thereof, gemcitabine and cisplatin are one week, two weeks, three weeks, one month, two months, three months, four months, five months, half a year or longer, respectively. Optionally, the time of each administration cycle is the same or different, and the interval between each administration cycle is the same or different.

12. A drug combination comprising an anti-PD-1 antibody or an antigen-binding fragment thereof, gemcitabine and cisplatin; preferably, the anti-PD-1 antibody or its antigen-binding fragment comprises a light chain complementary determining region with an amino acid sequence as shown in SEQ ID NOs: 1, 2 and 3, and a heavy chain complementary determining region with an amino acid sequence as shown in SEQ ID NOs: 4, 5 and 6; preferably, the anti-PD-1 antibody or its antigen-binding fragment comprises a light chain variable region with an amino acid sequence as shown in SEQ ID NO: 7, and a heavy chain variable region with an amino acid sequence as shown in SEQ ID NO: 8; preferably, the anti-PD-1 antibody comprises a light chain with an amino acid sequence as shown in SEQ ID NO: 9, and a heavy chain with an amino acid sequence as shown in SEQ ID NO: 10; more preferably, the anti-PD-1 antibody is toripalimab.

13. A method for preventing or treating a malignant tumor, the method comprising administering to an individual in need thereof: Administering an effective amount of the anti-PD-1 antibody or antigen-binding fragment thereof as claimed in any one of claims 5 or 6; preferably, the dosage and regimen of the anti-PD-1 antibody or antigen-binding fragment thereof is as described in any one of claims 7-11; or Administering the drug combination of claim 12; preferably, administering the anti-PD-1 antibody or antigen-binding fragment thereof, gemcitabine and cisplatin in the drug combination at a dosage or regimen as described in any one of claims 7 to 11; Preferably, the malignant tumor is nasopharyngeal carcinoma, more preferably recurrent or metastatic nasopharyngeal carcinoma, or the malignant tumor is as described in claim 3 or 4.

14. Use of a reagent for detecting gene mutation or amplification in the chromosome 11q13 region of CCND1, FGF14, FGF3 or FGF4 in an individual's peripheral blood circulating tumor DNA and / or tumor tissue and / or a reagent for detecting the number of EBV DNA copies in an individual's peripheral blood in the preparation of a kit for predicting the effect of an anti-PD-1 antibody or an antigen-binding fragment thereof in the treatment of nasopharyngeal carcinoma; preferably, the anti-PD-1 antibody or an antigen-binding fragment thereof is as described in claim 5 or 6.

15. A kit comprising: One or more single pharmaceutical dosage units of an anti-PD-1 antibody or an antigen-binding fragment thereof, the anti-PD-1 antibody or an antigen-binding fragment thereof as described in any one of claims 5 or 6; or One or more single drug dosage units of an anti-PD-1 antibody or an antigen-binding fragment thereof, one or more single drug dosage units of gemcitabine, and one or more single drug dosage units of cisplatin; preferably, the anti-PD-1 antibody or an antigen-binding fragment thereof is as described in any one of claims 5 or 6; or One or more single pharmaceutical dosage units of the pharmaceutical combination of claim 12.