Medications and methods for treating patients with completely resected mucosal melanoma.

Abstract

This invention relates to medicaments and methods for treating patients with completely resected mucosal melanoma, specifically to the use of an anti-PD-1 antibody or its antigen-binding fragment in the preparation of medicaments for treating patients with completely resected mucosal melanoma or for preventing recurrence or distant metastasis of mucosal melanoma in such patients. The invention also relates to a kit for treating patients with completely resected mucosal melanoma or for preventing recurrence or distant metastasis of mucosal melanoma in such patients. The medicaments and methods of this invention significantly prolong the remission survival time (RFS) in PDL1-positive patients and are well-tolerated.

Description

Technical Field

[0001] This invention relates to medicaments and methods for treating patients with completely resected mucosal melanoma, and more particularly to the use of an anti-PD-1 antibody or its antigen-binding fragment in the preparation of medicaments for treating patients with completely resected mucosal melanoma or for preventing recurrence or distant metastasis of mucosal melanoma in such patients. Background Technology

[0002] Although mucosal melanoma is a rare subtype of melanoma in Western countries, accounting for only 1.4% of all melanomas (Lian B, Cui CL, Zhou L, et al. The natural history and patterns of metastases from mucosal melanoma: an analysis of 706 prospectively-followed patients. Ann Oncol 2017 28:868-873), it ranks second among all melanoma subtypes in Asia, accounting for approximately 22.6% of such cases (Cui C, Lian B, Zhou L, et al. Multifactorial Analysis of Prognostic Factors and Survival Rates Among 706 Mucosal Melanoma Patients. Ann Surg Oncol 2018 25:2184-2192). Compared to cutaneous melanoma, mucosal melanoma has a very poor prognosis. Previous studies have shown that the 5-year overall survival (OS) rate for cutaneous melanoma is approximately 50-80%, but for mucosal melanoma it is only 25% (Lian B, Cui CL, Zhou L, et al. The natural history and patterns of metastases from mucosal melanoma: an analysis of 706 prospectively-followed patients. Ann Oncol 2017 28:868-873; Cui C, Lian B, Zhou L, et al. Multifactorial Analysis of Prognostic Factors and Survival Rates Among 706 Mucosal Melanoma Patients. Ann Surg Oncol 2018 25:2184-2192).Furthermore, early-stage mucosal melanoma is prone to recurrence after resection. A previous study showed that the median recurrent survival (RFS) in patients with resected mucosal melanoma was only 5.4 months (Lian B, SiL, Cui C, et al. Phase II randomized trial comparing high-dose IFNalpha2b with temozolomide plus cisplatin as systemic adjuvant therapy for resected mucosal melanoma. Clin Cancer Res 2013 19:4488-4498). Therefore, there remains an unmet medical need for systemic adjuvant therapy for resected mucosal melanoma.

[0003] Toripalimab (or JS001) is a humanized IgG4 monoclonal antibody (mAb) against programmed cell death receptor 1 (PD-1). It has been approved for second-line treatment of metastatic melanoma and metastatic urothelial carcinoma, and in China for third-line treatment of recurrent or metastatic nasopharyngeal carcinoma (Mai HQ, Chen QY, Chen D, et al. Toripalimab or placebo plus chemotherapy as first-line treatment in advanced nasopharyngeal carcinoma: a multicenter randomized phase 3 trial. NatMed 2021 27:1536-1543). In a previous phase Ib study, toripalimab in combination with the vascular endothelial growth factor receptor inhibitor axitinib showed good antitumor activity and can be used as first-line treatment for metastatic mucosal melanoma.

[0004] Previous clinical trials have shown that high-dose interferon (HDI)(IFN)-α2b can prolong relapse-free survival (RFS) and / or overall survival (OS) in high-risk cutaneous melanoma patients (Jonasch E, Kumar UN, Linette GP, et al. Adjuvant high-dose interferon alfa-2b in patients with high-risk melanoma. Cancer J 2000 6:139-145; Kirkwood JM, Ibrahim JG, Sosman JA, et al. High-dose interferon alfa-2b significantly prolongs relapse-free and overall survival compared with the GM2-KLH / QS-21 vaccine in patients with resectedstage IIB-III melanoma: results of intergroup trial E1694 / S9512 / C509801. J Clin Oncol 2001 19:2370-2380). In addition, several phase III clinical trials of cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) and PD-1 monoclonal antibodies have demonstrated their efficacy as adjuvant therapy for resected high-risk cutaneous melanomas (Eggermont AMM, Blank CU, Mandala M, et al. Adjuvant Pembrolizumab versus Placebo in Resected Stage III Melanoma. N Engl J Med 2018 378:1789-1801; Weber J, Mandala M, Del Vecchio M, et al. Adjuvant Nivolumab versus Ipilimumab in Resected Stage III or IV Melanoma. N Engl J Med 2017 377: 1824-1835). Although high-dose interferon is currently the only approved treatment for adjuvant melanoma, it is not universally accepted as a standard of care.The mounting evidence surrounding its questionable survival benefits, high incidence of serious toxicity, and negligible benefits for patients with more prevalent diseases makes it an unattractive treatment for most patients and clinicians (Schuchter L. Adjuvant Interferon Therapy for Melanoma: High-Dose, Low-Dose, No-Dose, Which-Dose J Clin Oncol 2004; 22:7-10). Therefore, there is a need for more effective treatments with acceptable safety profiles in adjuvant therapy. Summary of the Invention

[0005] This invention provides the use of an anti-PD-1 antibody or an antigen-binding fragment thereof in the preparation of a medicament for treating a patient with completely resected mucosal melanoma or for preventing recurrence or distant metastasis of mucosal melanoma in such a patient.

[0006] In another aspect, the present invention provides a method for adjuvant treatment of a patient with completely resected mucosal melanoma or for prevention of recurrence or distant metastasis of mucosal melanoma in such a patient, comprising administering to the patient an effective amount of the anti-PD-1 antibody of the present invention or an antigen-binding fragment thereof or a pharmaceutical composition thereof.

[0007] In another aspect, the present invention provides an anti-PD-1 antibody or an antigen-binding fragment thereof for adjuvant treatment of a patient with completely resected mucosal melanoma or for prevention of recurrence or distant metastasis of the mucosal melanoma in such a patient.

[0008] In one or more embodiments, the anti-PD-1 antibody or its antigen-binding fragment described in this invention is any antibody or antigen-binding fragment that can specifically bind to PD-1 and block the binding of PD-1 to its ligand PD-L1.

[0009] In one or more embodiments, the mucosal melanoma of the present invention is a head and neck mucosal melanoma or a non-head and neck mucosal melanoma.

[0010] In one or more embodiments, the mucosal melanoma of the present invention is a mucosal melanoma that is positive for PD-L1 expression in immunohistochemical staining analysis of tumor tissue sections or a mucosal melanoma that is negative for PD-L1 expression in immunohistochemical staining analysis of tumor tissue sections; preferably, it is a mucosal melanoma that is positive for PD-L1 expression in immunohistochemical staining analysis of tumor tissue sections.

[0011] In one or more embodiments, the anti-PD-1 antibody or its antigen-binding fragment of the present invention comprises a light chain complementarity-determining region as shown in SEQ ID NO: 1, 2 and 3, and a heavy chain complementarity-determining region as shown in SEQ ID NO: 4, 5 and 6.

[0012] In one or more embodiments, the anti-PD-1 antibody or its antigen-binding fragment of the present invention 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.

[0013] In one or more embodiments, the anti-PD-1 antibody of the present invention 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.

[0014] 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.

[0015] In one or more embodiments, the mucosal melanoma of the present invention is a head and neck mucosal melanoma, and the use further comprises administering one or more therapies to an individual in need; preferably, the therapy is radiotherapy.

[0016] In one or more embodiments, the dosage of the anti-PD-1 antibody or its antigen-binding fragment described in this invention is from about 0.1 mg / kg to about 10.0 mg / kg of individual body weight, for example, 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, for example, a fixed dose of about 120 mg, 240 mg, 360 mg, or 480 mg, preferably about 3 mg / kg of individual body weight or about 240 mg of fixed dose.

[0017] In one or more embodiments, the anti-PD-1 antibody or its antigen-binding fragment 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.

[0018] In one or more embodiments, the dosage of the anti-PD-1 antibody or its antigen-binding fragment described in this invention is 1 mg / kg body weight, 3 mg / kg body weight, 10 mg / kg body weight, or a fixed dose of 240 mg or 480 mg, administered once every two or three weeks.

[0019] In one or more embodiments, the anti-PD-1 antibody or its antigen-binding fragment of the present invention is administered at a dose of 3 mg / kg individual body weight or a fixed dose of 240 mg every two weeks.

[0020] In one or more embodiments, the anti-PD-1 antibody or its antigen-binding fragment of the present invention is administered via a parenteral route, such as intravenous infusion, in a liquid dosage form, such as an injection.

[0021] In one or more embodiments, the administration cycle of the anti-PD-1 antibody or its antigen-binding fragment according to the present invention is one week, two weeks, three weeks, one month, two months, three months, four months, five months, six months, one year or longer. Optionally, the duration of each administration cycle may be the same or different, and the interval between each administration cycle may be the same or different.

[0022] In one or more embodiments, the radiotherapy dose of the present invention is CTV. TB 60–64 Gy / 30 times, administered on days 1–5 of each week for a total of 6 weeks; preferably, radiotherapy is administered concurrently with the administration of anti-PD-1 antibody or its antigen-binding fragment for 6–8 weeks.

[0023] In another aspect, the present invention provides a combination therapy comprising administering to a patient in need an anti-PD-1 antibody or an antigen-binding fragment thereof or a pharmaceutical composition thereof as described herein and one or more additional therapies.

[0024] In one or more embodiments, the additional therapy described in this invention is selected from chemotherapeutic agents, biological agents, immunogenic agents, immunostimulatory cytokines, encoded immunostimulatory cytokines, or radiotherapy.

[0025] In one or more embodiments, the pharmaceutical composition of the present invention comprises the anti-PD-1 antibody or its antigen-binding fragment described herein, and a pharmaceutically acceptable carrier or excipient.

[0026] In one or more embodiments, the method of the present invention has fewer grade 3 or higher treatment-related adverse events (TEAEs) than high-dose interferon alpha-2b (HDI) adjunctive therapy. Preferably, the grade 3 or higher treatment-related adverse events (TEAEs) are about 60%, 50%, 40%, 30%, 20%, or 10% lower than HDI.

[0027] In one or more embodiments, the method of the present invention has fewer grade 3 or higher treatment-related adverse events (TEAEs) than high-dose interferon α2b (HDI) adjunctive therapy. Preferably, the grade 3 or higher treatment-related adverse events (TEAEs) are about 10%-60%, 10%-50%, 10%-40%, 10%-30%, or 10%-20% lower than HDI.

[0028] In one or more embodiments, the adjuvant therapy described in this invention is more effective than high-dose interferon α-2b (HDI) adjuvant therapy. Preferably, the relapse-free survival (RFS) of PD-L1-positive patients is about 50%, 40%, 30%, 20%, or 10% higher than that of HDI.

[0029] In one or more embodiments, the adjuvant therapy described in this invention is more effective than high-dose interferon α-2b (HDI) adjuvant therapy. Preferably, the relapse-free survival (RFS) of PD-L1-positive patients is about 10-50%, 10-40%, 10-30%, or 10-20% higher than that of HDI.

[0030] In another aspect, the present invention provides a kit comprising one or more single-dose units of an anti-PD-1 antibody or an antigen-binding fragment thereof or a pharmaceutical composition thereof, wherein the anti-PD-1 antibody or the antigen-binding fragment thereof is as described in any of the present invention. The kit of the present invention can be used to treat patients with completely resected mucosal melanoma or to prevent recurrence or distant metastasis of mucosal melanoma in such patients.

[0031] In another aspect, the present invention provides a method for predicting the efficacy of anti-PD-1 antibody or its antigen-binding fragment in treating a patient with completely resected mucosal melanoma to prevent recurrence or distant metastasis of the mucosal melanoma. Attached Figure Description

[0032] Figure 1 Kaplan-Meier curves for recurrence-free survival (RFS) of the entire analyzed population.

[0033] Figure 2 : Hazard ratio (HR) of recurrence-free survival (RFS) in each subgroup.

[0034] Figure 3 Overall survival (OS) hazard ratio (HR) for each subgroup.

[0035] Figure 4: Kaplan-Meier curves of recurrence-free survival (RFS) for subgroups: A) RFS of patients with PD-L1 positive tumors; B) RFS of patients with PD-L1 negative tumors.

[0036] Figure 5 Current research flowchart. Detailed Implementation

[0037] This invention relates to a treatment method for the complete resection of mucosal melanoma. The method of this invention includes administering the anti-PD-1 antibody or its antigen-binding fragment described herein to a patient in need.

[0038] the term

[0039] To facilitate understanding of this invention, certain technical terms are specifically defined below. Unless otherwise expressly stated elsewhere in this document, the technical terms used herein have the meanings commonly understood by one of ordinary skill in the art to which this invention pertains.

[0040] "Administration," "giving," and "treatment" refer to introducing a composition containing a therapeutic agent into a subject using any of the various methods or delivery systems known to those skilled in the art. Routes of administration for anti-PD-1 antibodies include intravenous, intramuscular, subcutaneous, peritoneal, spinal, or other parenteral routes, such as injection or infusion. "Parenteral administration" refers to administration by injection other than enteral or local administration, including but not limited to intravenous, intramuscular, intra-arterial, intrathecal, intralymphatic, intra-tracheal, intracapsular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, intra-articular, and intrasternal injections and infusions, as well as intracorporeal electroporation.

[0041] As used herein, an “adverse reaction” (AE) is any unfavorable and usually unintentional or undesirable sign, symptom, or illness associated with the use of medical treatment. For example, an adverse reaction may be associated with activation of the immune system or expansion of immune system cells in response to treatment. Medical treatment may have one or more associated AEs, and each AE may have the same or different levels of severity.

[0042] "Tumor burden" refers to the total amount of tumor material distributed throughout the body. It refers to the total number of cancer cells or the total size of the tumor. Tumor burden can be measured using various methods known in the art, such as using calipers after tumor removal from the subject, or measuring its size while in vivo using imaging techniques (such as ultrasound, bone scans, computational tomography (CT), or magnetic resonance imaging (MRI) scans).

[0043] The term "tumor size" refers to the total size of a tumor, which can be measured as its length and width. Tumor size can be determined using a variety of methods known in the art, such as using calipers after the tumor has been removed from the subject, or using imaging techniques (such as bone scans, ultrasound, CT, or MRI scans) while the tumor is in vivo.

[0044] The terms “subject,” “individual,” and “object” include any living organism, preferably an animal, more preferably a mammal (e.g., rat, mouse, dog, cat, rabbit, etc.), and most preferably a human being. The terms “subject” and “patient” are used interchangeably herein.

[0045] The term "antibody" as used herein refers to any form of antibody capable of achieving the desired biological 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 camel-derived single-domain antibodies. An "antibody" specifically binds to an antigen and comprises at least two heavy (H) chains and two light (L) chains linked by disulfide bonds. Each heavy chain contains a heavy chain variable region (VH) and a heavy chain constant region, the heavy chain constant region containing three constant domains CH1, CH2, and CH3. Each light chain contains a light chain variable region (VL) and a light chain constant region, the light chain constant region containing one constant domain CL. The VH and VL regions can be further subdivided into hypervariable regions called complementarity-determining regions (CDRs), which are dispersed within more conserved regions called framework regions (FRs). Generally, from the N-terminus to the C-terminus, both the light chain and heavy chain variable domains contain 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.

[0046] The carboxyl terminus of the heavy chain can define a constant region primarily responsible for effector function. Typically, human light chains are divided into κ and λ chains. Human heavy chains are usually classified into μ, δ, γ, α, or ε chains, and antibody isotypes are defined 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.

[0047] 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 through recombinant methods to reduce their immunogenicity in humans.

[0048] Unless otherwise expressly stated, “antibody fragment” or “antigen-binding fragment” as used herein refers to an antigen-binding fragment of an antibody, that is, an antibody fragment that retains the specific ability of the full-length antibody to bind to an antigen, such as a fragment retaining one or more CDR regions. Examples of antigen-binding fragments include, but are not limited to, Fab, Fab', F(ab')2, and Fv fragments; double-chain antibodies; linear antibodies; single-chain antibody molecules; nanobodies; and multispecific antibodies formed from antibody fragments.

[0049] "Chimeric antibody" refers to an antibody and its fragments in which a portion of the heavy chain and / or light chain is identical or homologous to the corresponding sequence in an antibody derived from a specific species (such as human) or belonging to a specific antibody class or subclass, while the remaining portion of the chain is identical or homologous to the corresponding sequence in an antibody derived from another species (such as mouse) or belonging to another antibody class or subclass, provided that it exhibits the desired biological activity.

[0050] "Human antibody" refers to an antibody that contains only human immunoglobulin sequences. If the human antibody is produced in a mouse, mouse cell, or hybridoma derived from mouse cells, it may contain mouse carbohydrate chains. Similarly, "mouse antibody" or "rat antibody" refers to an antibody that contains only mouse or rat immunoglobulin sequences, respectively.

[0051] "Humanized antibodies" refer to antibody forms containing sequences derived from non-human (e.g., mouse) antibodies and human antibodies. These antibodies contain a minimal sequence derived from a single side of a non-human immunoglobulin. Typically, humanized antibodies will contain substantially all of at least one and usually two variable domains, wherein all or substantially all of the hypervariable loops correspond to the hypervariable loops of the non-human immunoglobulin, and all or substantially all of the FR regions are the FR regions of the human immunoglobulin. Humanized antibodies optionally also include at least a portion of the immunoglobulin constant region (Fc) (usually the human immunoglobulin constant region).

[0052] The term "immunotherapy" refers to the treatment of a subject who has a disease or is at risk of infection or relapse of disease by means of methods including inducing, enhancing, suppressing, or otherwise modifying an immune response. "Treatment" or "therapy" for a subject means any type of intervention or procedure performed on a subject, or the administration of an active agent to a subject with the aim of reversing, alleviating, improving, slowing, or preventing the onset, progression, severity, or relapse of symptoms, complications, or conditions, or disease-related biochemical markers.

[0053] "Programmed death receptor-1 (PD-1)" refers to an immunosuppressive 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. As used herein, the term "PD-1" includes human PD-1 (hPD-1), variants, isotypes, and species homologs of hPD-1, as well as analogs that share at least one common epitope with hPD-1.

[0054] A "therapeutic effective amount" or "therapeutic 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 remission, demonstrated by a reduction in the severity of disease symptoms, an increase in the frequency and duration of asymptomatic periods of disease, or prevention of injury or disability caused by disease suffering. The ability of a therapeutic agent to promote disease remission 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 predicting human efficacy, or by determining the activity of the drug in an in vitro assay.

[0055] Therapeutic effective doses of a drug include “preventive effective doses,” which are any amount of drug that inhibits the development or recurrence of cancer when administered alone or in combination with an antitumor agent to a subject at risk of developing cancer or a subject with a recurrence of cancer.

[0056] "Biotherapeutic agents" refer to biomolecules, such as antibodies or fusion proteins, that block ligand / receptor signaling in any biological pathway that supports tumor maintenance and / or growth or inhibits antitumor immune responses.

[0057] Unless otherwise expressly stated, “CDR” as used herein refers to the immunoglobulin variable region being the complementarity-determining region as defined using the Kabat numbering system.

[0058] "Therapeutic anti-PD-1 monoclonal antibody" refers to an antibody that specifically binds to the mature form of a particular form of PD-1 expressed on the surface of certain mammalian cells. Mature PD-1 lacks a prosecuratory leader sequence, or leader peptide. The terms "PD-1" and "mature PD-1" are used interchangeably in this document and should be understood as the same molecule unless otherwise clearly defined or evident from the context.

[0059] As described in this article, therapeutic anti-human PD-1 antibodies or anti-hPD-1 antibodies refer to monoclonal antibodies that specifically bind to mature human PD-1.

[0060] The “frame region” or “FR” mentioned in this article refers to the immunoglobulin variable region excluding the CDR region.

[0061] "Isolated antibody or its antigen-binding fragment" refers to a purified state in which the specified molecule does not substantially contain other biomolecules, such as nucleic acids, proteins, lipids, carbohydrates or other materials (such as cell debris or growth medium).

[0062] "Patient," "patient," or "subject" means any single subject who requires medical intervention or participates in a clinical trial, epidemiological study, or is used as a control, and is typically a mammal, including humans and other mammals such as horses, cows, dogs, or cats.

[0063] The “RECIST 1.1 efficacy criteria” described in this article refer to the definition given by Eisenhauver et al. and EA et al., Eur. J Cancer 45:228-247 (2009), based on the context of the measured response and targeting or non-target damage. Prior to immunotherapy, it was the most commonly used standard for evaluating the efficacy of treatments in solid tumors. However, with the advent of the immunotherapy era, many challenges have arisen that were previously unseen in tumor evaluation. Therefore, based on the newly emerging phenomena arising from immunotherapy itself, in 2016, the RECIST working group revised the existing “RECIST v.1.1” and proposed a new set of criteria, namely the “irRECIST criteria” described in this article, aiming to better assess the efficacy of immunotherapy drugs.

[0064] The term "ECOG" (Energy Performance Status) scoring system is an indicator of a patient's general health status and tolerance to treatment based on their physical ability. The ECOG performance status scoring system uses a scale of 0, 1, 2, 3, 4, and 5. A score of 0 indicates completely normal activity levels, with no difference from pre-illness activity levels. A score of 1 indicates the ability to walk freely and perform light physical activities, including general housework or office work, but not to engage in strenuous physical activity.

[0065] "Sustained response" refers to a sustained therapeutic effect after discontinuation of the therapeutic agent or combination therapy described herein. In some embodiments, the sustained response has a duration at least equal to or at least 1.5, 2.0, 2.5, or 3 times the duration of treatment.

[0066] A "tissue slice" refers to a single portion or slice of a tissue sample, such as a thin slice of tissue cut from a sample of normal tissue or a tumor.

[0067] The term "treatment" for cancer as used herein refers to the use of the treatment regimens described herein (such as administration of anti-PD-1 antibodies) in subjects who have cancer or have been diagnosed with cancer to achieve at least one positive therapeutic effect (e.g., a reduction in the number of cancer cells, a decrease in tumor volume, a reduction in the rate of cancer cell invasion into surrounding organs, or a reduction in the rate of tumor metastasis or growth). Positive therapeutic effects in cancer can be measured in various ways (see WA Weber, J. Nucl. Med., 50: 1S-10S (2009)). For example, regarding tumor growth inhibition, according to NCI criteria, T / C ≤ 42% is the minimum level of antitumor 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 when cancer does not grow, including the amount of time a patient experiences 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 this invention, which effectively treats cancer patients, can vary depending on various factors, such as the patient's disease state, age, weight, and the ability of the therapy to elicit an anti-cancer response in the subject. Although embodiments of this invention may not achieve effective positive therapeutic effects in every subject, they should be effective and achieve positive therapeutic effects in a statistically significant number of subjects.

[0068] The terms "method of administration" and "administration regimen" are used interchangeably and refer to the dosage and timing of each therapeutic agent in the combination of the present invention.

[0069] The term "immunohistochemistry (IHC)" refers to a method that utilizes the principle of specific binding between antigens and antibodies to determine intracellular antigens (peptides and proteins) by using a chemical reaction to develop color in labeled antibody chromogenic agents (fluorescein, enzymes, metal ions, isotopes), and to study their localization, qualitative analysis, and relative quantification. In some embodiments of the present invention, before treatment with anti-PD-1 antibodies, tumor tissue samples from the subject are subjected to PD-L1 detection using Roche's anti-human PD-L1 antibody SP142 (Cat No: M4422). In some embodiments, a membrane staining intensity of ≥1% for tumor cells is defined as PD-L1 positive.

[0070] In this document, the term "cancer" or "malignant tumor" refers to a wide variety of diseases characterized by the uncontrolled growth of abnormal cells in the body. Unregulated cell division, growth, and proliferation lead to the formation of malignant tumors, which invade adjacent tissues and can 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, medicines, and kits of this invention include, but are not limited to, carcinoma, lymphoma, leukemia, blastoma, and sarcoma. More specific examples of cancer include squamous cell carcinoma, myeloma, small cell lung cancer, non-small cell lung cancer, glioma, Hodgkin lymphoma, non-Hodgkin lymphoma, acute myeloid leukemia, multiple myeloma, gastrointestinal (intestinal) 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 carcinoma, cervical cancer, brain cancer, stomach cancer, bladder cancer, hepatocellular carcinoma, breast cancer, colon cancer, and head and neck cancer.

[0071] In this document, 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 invention, the tumor mutation burden (TMB) is estimated by analyzing somatic mutations, including coding base substitutions and large base insertions in the panel sequence under study.

[0072] The various aspects of the invention are further described in detail in the following paragraphs.

[0073] Anti-PD-1 antibody

[0074] In this document, "PD-1 antibody" refers to any chemical compound or biomolecule that binds to the PD-1 receptor, blocking the binding of PD-L1 expressed on cancer cells to PD-1 expressed on immune cells (T, B, NK cells), and preferably also blocking the binding of PD-L2 expressed on cancer cells to PD-1 expressed on immune cells. Alternative names or synonyms for PD-1 and its ligands include: for PD-1, PDCD1, PD1, CD279, and SLEB2; for PD-L1, PDCD1L1, PDL1, B7-H1, B7H1, B7-4, CD274, and B7-H; and for PD-L2, PDCD1L2, PDL2, B7-DC, and CD273. In any treatment method, medicine, or use of this invention for treating a human individual, 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 number NP_005009. The amino acid sequences of human PD-L1 and PD-L2 can be found at NCBI loci number NP_054862 and NP_079515, respectively.

[0075] In this article, when “anti-PD-1 antibody” is mentioned, unless otherwise stated or described, the term includes its antigen-binding fragment.

[0076] Anti-PD-1 antibodies applicable to any of the uses, therapies, pharmaceuticals, and kits described herein bind to PD-1 with high specificity and high affinity, blocking the binding of PD-L1 / 2 to PD-1 and inhibiting PD-1 signal transduction, thereby achieving an immunosuppressive effect. In any of the uses, therapies, pharmaceuticals, and kits disclosed herein, the anti-PD-1 antibody includes the full-length antibody itself, and an antigen-binding portion or fragment that binds to the PD-1 receptor and exhibits functional properties similar to a full-length antibody 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 to bind to human PD-1. In other embodiments, the anti-PD-1 antibody or its antigen-binding fragment is a chimeric, humanized, or human antibody or its antigen-binding fragment. In some embodiments used to treat human individuals, the antibody is a humanized antibody.

[0077] In some embodiments, the anti-PD-1 antibody for any use, therapy, medicament, or kit described in this invention 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 may be a human antibody, a humanized antibody, or a chimeric antibody, and may include a human constant region. In some embodiments, the constant region is selected from the group consisting of the constant regions of human IgG1, IgG2, IgG3, and IgG4; preferably, the anti-PD-1 antibody or its antigen-binding fragment suitable for any use, therapy, medicament, or kit described in this invention comprises a heavy chain constant region of human IgG1 or IgG4 isotypes, 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 its antigen-binding fragment includes an S228P mutation, which replaces a serine residue in the hinge region with a proline residue normally present at the corresponding position on an IgG1 isotype antibody.

[0078] Preferably, in any embodiment of the uses, therapies, drugs and kits described in this invention, the PD-1 antibody is a monoclonal antibody or its antigen-binding fragment, wherein its light chain CDR is the amino acid shown in SEQ ID NO: 1, 2 and 3, and its heavy chain CDR is the amino acid shown in SEQ ID NO: 4, 5 and 6.

[0079] More preferably, in any embodiment of the uses, therapies, drugs and kits described in this 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.

[0080] More preferably, in any embodiment of the uses, therapies, drugs and kits described in this 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.

[0081] Table A provides the amino acid sequence numbers of the light chain CDR and heavy chain CDR of exemplary anti-PD-1 antibody mAbs used in the purposes, therapies, pharmaceuticals, and kits described in this invention, numbered according to Kabat rules:

[0082] Table A: Light and Heavy Chain CDRs of Exemplary Anti-Human PD-1 Antibodies

[0083] LCDR1 SEQ ID NO: 1 LCDR2 SEQ ID NO: 2 LCDR3 SEQ ID NO: 3 HCDR1 SEQ ID NO: 4 HCDR2 SEQ ID NO: 5 HCDR3 SEQ ID NO: 6 VL SEQ ID NO: 7 VH SEQ ID NO: 8 LC SEQ ID NO: 9 HC SEQ ID NO: 10

[0084] Examples of anti-PD-1 antibodies that bind to human PD-1 and can be used for the purposes, therapies, medicines and kits described in this invention are set forth in WO2014206107. Human PD-1 mAbs that can be used as anti-PD-1 antibodies in the uses, therapies, medicines, and kits described in this invention include any one of the anti-PD-1 antibodies described in WO2014206107, including: Toripalimab (a humanized IgG4 mAb having the structure described in WHO Drug Information (Vol. 32, No. 2, pp. 372-373 (2018)) and comprising the light and heavy chain amino acid sequences shown in SEQ ID NO: 9 and 10. In a preferred embodiment, the anti-PD-1 antibody that can be used in any of the uses, therapies, medicines, and kits described in this invention is selected from humanized antibodies 38, 39, 41, and 48 described in WO2014206107. In a particularly preferred embodiment, the anti-PD-1 antibody that can be used in any of the uses, therapies, medicines, and kits described in this invention is toripalimab.

[0085] Anti-PD-1 antibodies that can be used for any of the uses, therapies, medicines and kits described in this invention also include FDA-approved Nivolumab and Pembrolizumab.

[0086] In some embodiments, anti-PD-1 antibodies that can be used for any of the purposes, therapies, pharmaceuticals and kits described in this invention 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.

[0087] As used herein, “PD-L1” expression or “PD-L2” expression refers to any detectable expression level of a specific PD-L protein on the cell surface 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 in tumor cells can be detected by PET imaging using a binder that specifically binds to the desired PD-L target (such as PD-L1 or PD-L2).

[0088] For methods used to quantify PD-L1 protein expression in IHC analysis of tumor tissue sections, see, but 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).

[0089] One approach uses a simple binary endpoint of PD-L1 expression positivity or negativity, where a positive result is defined as the percentage of tumor cells showing histological evidence of cell surface membrane staining. PD-L1 expression positivity is defined as a tumor tissue section counted at more than 1% of total tumor cells.

[0090] In another method, PD-L1 expression in tumor tissue sections is quantified in tumor cells and in infiltrating immune cells. The percentage of membrane-stained tumor cells and infiltrating immune cells is quantified separately as ≤1%, 1% to 50%, and subsequently 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%.

[0091] In some implementations, the PD-L1 expression level in malignant cells and / or infiltrating immune cells within the tumor is determined as "overexpressed" or "elevated" based on a comparison with the PD-L1 expression level of an appropriate control. For example, the protein or mRNA expression level of the control PD-L1 may be the level quantified in the same type of non-malignant cells or in sections from matched normal tissue.

[0092] In some implementations, PD-L1 positivity is defined as ≥1% CPS of tumor cells stained with JS311 IHC. CPS represents the positive composite score. The CPS criterion is: the percentage of tumor cells stained with membranes of any intensity, and the percentage of lymphocytes / macrophages stained with membranes / cytoplasm directly associated with tumor cells relative to tumor cells (at least one hundred), but excluding all necrotic cells, stromal cells, carcinoma in situ, and other immune cells (including but not limited to neutrophils, eosinophils, and plasma cells).

[0093] adjunctive therapy

[0094] As used herein, “adjuvant therapy” or “adjuvant treatment” is understood to be the administration of one or more medications to a patient after surgical removal of one or more cancerous tumors in which all detectable and resectable disease (e.g., cancer) has been removed from the patient, but there is a statistical risk of recurrence due to occult lesions, with the aim of reducing the likelihood or severity of recurrence or delaying the appearance of biological manifestations of disease recurrence.

[0095] Effective dose and excision

[0096] The term "effective amount" refers to the amount of a drug or agent that will elicit a biological or medical response in a tissue, system, animal, or human, as sought by, for example, an investigator or clinician. Furthermore, the term "therapeuticly effective amount" refers to any amount that, compared to a corresponding subject who has not received that amount, results in treatment, cure, prevention, or relief of disease, condition, or side effects, or a reduction in the rate of disease or condition progression. Within its scope, the term also includes amounts that effectively enhance normal physiological function. Therapeuticly effective amounts of drugs also include "preventatively effective amounts."

[0097] Regarding solid tumors, an effective amount includes an amount sufficient to cause tumor shrinkage and / or reduce the rate of tumor growth (such as inhibiting tumor growth) or delay other unwanted cell proliferation. In some embodiments, an effective amount is an amount sufficient to prevent or delay tumor recurrence. An effective amount may be administered in one or more doses. An effective amount of a drug or composition may: (i) reduce the number of cancer cells; (ii) reduce tumor size; (iii) inhibit, delay, or slow down cancer cells to a certain extent and prevent cancer cell invasion into peripheral organs; (iv) inhibit, i.e., slow down, to a certain extent and prevent tumor metastasis; (v) inhibit tumor growth; (vi) prevent or delay the occurrence and / or recurrence of tumors; and / or (vii) alleviate one or more cancer-related symptoms to a certain extent.

[0098] The term "resection" refers to the surgical removal of the characteristic malignant tissue of a melanoma from a human patient. Resection can be understood as the removal of malignant tissue so that the presence of any remaining malignant tissue in the patient is undetectable by existing methods. Resection can be understood as the removal of melanoma so that the presence of any remaining melanoma in the patient is undetectable. Resection may include complete resection or partial resection.

[0099] Recurrence and distant metastasis

[0100] The term "recurrence" refers to a local recurrence of a tumor or melanoma, meaning that in a patient who has had a tumor or melanoma removed / completely removed, the tumor or melanoma reappears within 2 cm of the original tumor site. Local recurrence of melanoma can occur at any location of the primary melanoma or in the nearest regional lymph nodes to the primary melanoma.

[0101] The term "distant metastasis" refers to the distant metastasis of a tumor or melanoma. This means that in patients who have had their tumor or melanoma removed or completely removed, the tumor or melanoma reappears at a distance of more than 2 cm from the original tumor area. At this time, the tumor or melanoma cells can spread to distant parts of the body, such as the lungs, liver, or other parts of the body.

[0102] Combination therapy, dosage and dosing regimen

[0103] The combination therapy of the present invention may also include one or more additional therapies. Additional therapeutic agents may be, for example, chemotherapeutic agents, biological 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), cells transfected with genes encoding immunostimulatory cytokines (such as, but not limited to, GM-CSF), or radiotherapy.

[0104] The selection of a dosing regimen (also referred to herein as an administration regimen) for the medicaments of the present invention depends on several factors, including the solid serum or tissue turnover rate of the treated individual, symptom levels, overall immunogenicity, and accessibility of target cells, tissues, or organs. Preferably, the dosing regimen maximizes the amount of each therapeutic agent delivered to the patient in accordance with acceptable levels of side effects. Therefore, the dosage and frequency of administration of each biological and chemotherapeutic agent depend in part on the specific therapeutic agent, the severity of the cancer being treated, and the patient's characteristics. Guidance can be obtained for selecting appropriate dosages of antibodies, cytokines, and small molecules. The determination of an appropriate dosing regimen can be made by a clinician, for example, by referring to parameters or factors known or suspected in the art to affect or expected to affect treatment, and will depend on, for example, the patient's clinical history (e.g., previous treatments), the type and stage of the cancer being treated, and biomarkers of response to one or more therapeutic agents in combination therapy.

[0105] Each therapeutic agent or therapy of the combination therapy of the present invention can be administered simultaneously (i.e., in the same pharmaceutical composition), concurrently (i.e., as separate pharmaceutical formulations, administered one after another in any order), or sequentially in any order. Sequential administration is particularly useful when the therapeutic agents in the combination therapy may be in different dosage forms (one drug is a tablet or capsule and the other is a sterile liquid formulation) and / or at different dosing schedules (e.g., a chemotherapeutic agent is administered at least daily and a biological agent is administered less frequently (e.g., once a week, once every two weeks, or once every three weeks)).

[0106] In some embodiments, at least one therapeutic agent in the combination therapy is administered using the same dosing regimen (treatment dose, frequency, and duration) typically used when the agent is used as a monotherapy to treat the same tumor. In other embodiments, the patient receives a smaller total amount of at least one therapeutic agent in the combination therapy compared to when the agent is used as a monotherapy, for example, a smaller dose, a less frequent dose, and / or a shorter treatment duration.

[0107] Each therapeutic agent in the combination therapy of the present invention can be administered orally or parenterally, including intravenous, intramuscular, intraperitoneal, subcutaneous, rectal, local, and percutaneous routes.

[0108] The anti-PD-1 antibody or its antigen-binding fragment of the present invention can be administered by continuous infusion or by intermittent dosing. Single-dose doses can range from about 0.01 mg / kg to about 20 mg / kg, from about 0.1 mg / kg to about 10 mg / kg body weight, or a fixed dose from about 120 mg to about 480 mg. For example, doses can be about 0.1 mg / kg, about 0.3 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, or about 10 mg / kg body weight. Alternatively, fixed doses of about 120 mg, 240 mg, 360 mg, or 480 mg can be administered. Dosing regimens are typically designed to achieve exposure that results in sustained receptor occupancy (RO) based on the typical pharmacokinetic characteristics of the antibody. Representative dosing regimens may include approximately once weekly, approximately once every two weeks, approximately once every three weeks, approximately once every four weeks, approximately once a month, or longer. In some implementation schemes, anti-PD-1 antibodies are administered to individuals approximately every two weeks.

[0109] In some embodiments, the anti-PD-1 antibody of the present invention is toripalimab, with a single administration dose selected from about 1 to about 5 mg / kg body weight. In some embodiments, the single administration dose of toripalimab is selected from doses of about 1 mg / kg, 2 mg / kg, 3 mg / kg, 3 mg / kg, 4 mg / kg, and 5 mg / kg body weight, or fixed doses of about 120 mg, 240 mg, and 360 mg, administered intravenously. In some preferred embodiments, toripalimab is administered as a liquid drug, with the selected dose administered via intravenous infusion over a period of 30 to 60 minutes. In some embodiments, toripalimab is administered at a fixed dose of about 3 mg / kg or about 240 mg every two weeks (Q2W) via intravenous infusion over a period of 30 minutes.

[0110] In some embodiments, the mucosal melanoma is a head and neck mucosal melanoma, and radiotherapy is administered concurrently with anti-PD-1 antibody or its antigen-binding fragment for 6-8 weeks. The radiotherapy dose is 60-64 Gy / 30 fractions of CTVTB, administered on days 1-5 of each week for a total of 6 weeks.

[0111] The administration cycles of the anti-PD-1 antibody or its antigen-binding fragment of the present invention and chemotherapy are 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. In some embodiments, one administration / treatment cycle is two weeks. In some embodiments, in one administration cycle, toripalimab is administered at approximately 3 mg / kg every two weeks. In some embodiments, one administration / treatment cycle of radiotherapy is one week. In some embodiments, in one administration cycle, the radiotherapy dose is CTVTB 60-64 Gy / 30 fractions, administered on days 1-5 of each week. The administration duration of the anti-PD-1 antibody or its antigen-binding fragment of the present invention is no more than one year. The administration duration of radiotherapy of the present invention is 6 weeks, administered concurrently within 6-8 weeks of the administration of the anti-PD-1 antibody or its antigen-binding fragment.

[0112] Treatment methods and uses

[0113] The purpose of this invention is to treat patients with completely resected mucosal melanoma and prevent recurrence or distant metastasis. This invention has found that administering an effective amount of any of the anti-PD-1 antibodies described herein or their antigen-binding fragments, or a pharmaceutical composition containing such anti-PD-1 antibodies or their antigen-binding fragments, can effectively prevent recurrence or distant metastasis of mucosal melanoma in patients with completely resected mucosal melanoma, improving recurrence-free survival (RFS) and / or distant metastasis-free survival (DMFS); simultaneously, it improves overall survival. In some embodiments, administering an effective amount of any of the anti-PD-1 antibodies described herein or their antigen-binding fragments, or a pharmaceutical composition containing such anti-PD-1 antibodies or their antigen-binding fragments, improves patient safety and tolerability.

[0114] Therefore, the present invention provides the use of the aforementioned anti-PD-1 antibody or its antigen-binding fragment in the preparation of a medicament for the prevention or treatment of patients with completely resected mucosal melanoma or for the prevention of recurrence or distant metastasis of mucosal melanoma in such patients.

[0115] The present invention also provides a method for preventing or treating a patient with completely resected mucosal melanoma, or for preventing recurrence or distant metastasis of mucosal melanoma in such a patient, comprising administering to the patient an effective amount of the anti-PD-1 antibody or its antigen-binding fragment thereof described in this invention. The effective amount includes both preventative and therapeutic effective amounts.

[0116] The present invention also provides the aforementioned anti-PD-1 antibody or antigen-binding fragment thereof for use in treating patients with completely resected mucosal melanoma or in preventing recurrence or distant metastasis of mucosal melanoma in such patients.

[0117] The present invention also provides the administration of therapeutically effective amounts of one or more therapies (e.g., treatment modalities and / or other therapeutic agents) to a subject in combination. In some embodiments, the therapies include surgical treatment and / or radiation therapy.

[0118] In this article, "complete resection of mucosal melanoma" refers to the complete removal of the primary lesion with negative surgical margins.

[0119] In some embodiments, the methods or uses provided by the present invention further include administering one or more therapies (e.g., treatment modalities and / or other therapeutic agents) to an individual in need. The antibodies of the present invention can be used alone or in combination with other therapeutic agents in a therapy. For example, they can be co-administered with at least one additional therapeutic agent. In some embodiments, the methods or uses provided by the present invention further include administering radiotherapy (radiotherapy) to an individual in need.

[0120] In one or more embodiments, the mucosal melanoma of the present invention is a head and neck mucosal melanoma or a non-head and neck mucosal melanoma.

[0121] In one or more embodiments, the mucosal melanoma of the present invention is a mucosal melanoma that is positive for PD-L1 expression in immunohistochemical staining analysis of tumor tissue sections or a mucosal melanoma that is negative for PD-L1 expression in immunohistochemical staining analysis of tumor tissue sections; preferably, it is a mucosal melanoma that is positive for PD-L1 expression in immunohistochemical staining analysis of tumor tissue sections.

[0122] Preferably, the method, use, or anti-PD-1 antibody described in any embodiment of the present invention is particularly suitable for mucosal melanomas that show positive PD-L1 expression in immunohistochemical staining analysis of tumor tissue sections.

[0123] Preferred anti-PD-1 antibodies for treating patients with completely resected mucosal melanoma or for preventing recurrence or distant metastasis of mucosal melanoma in such patients may be as described in any embodiment herein, more preferably antibodies with light chain CDRs of the amino acids shown in SEQ ID NO: 1, 2 and 3 and heavy chain CDRs of the amino acids shown in SEQ ID NO: 4, 5 and 6, more preferably monoclonal antibodies containing the light chain variable region shown in SEQ ID NO: 7 and the heavy chain variable region shown in SEQ ID NO: 8, more preferably monoclonal antibodies containing the light chain shown in SEQ ID NO: 9 and the heavy chain shown in SEQ ID NO: 10, more preferably humanized antibodies 38, 39, 41 and 48 described in WO2014206107, and most preferably toripalimab.

[0124] In a particularly preferred embodiment, the present invention provides a method for treating a patient with completely resected mucosal melanoma or for preventing recurrence or distant metastasis of the mucosal melanoma in such a patient, the method comprising administering the patient a therapeutically or preventively effective amount of toripalimab; preferably, the patient is PD-L1 positive. In some embodiments, the mucosal melanoma is a non-head and neck mucosal melanoma. In some embodiments, the mucosal melanoma is a head and neck mucosal melanoma, and the method further comprises concurrent administration of radiotherapy within 6-8 weeks of toripalimab administration.

[0125] In a particularly preferred embodiment, the present invention provides the use of an anti-PD-1 antibody or an antigen-binding fragment thereof in the preparation of a medicament for treating a patient with completely resected mucosal melanoma or for preventing recurrence or distant metastasis of mucosal melanoma in such a patient. Preferably, the tumor tissue sections from the patient with completely resected mucosal melanoma show positive PD-L1 expression (CPS ≥ 1%) in immunohistochemical staining analysis. In some embodiments, the mucosal melanoma is a non-head and neck mucosal melanoma. In some embodiments, the mucosal melanoma is a head and neck mucosal melanoma, and the use further includes concurrent administration of radiotherapy within 6-8 weeks of toripalimab administration.

[0126] medicine box

[0127] The present invention also provides a kit containing one or more single-dose units of an anti-PD-1 antibody or an antigen-binding fragment thereof as described in any embodiment herein.

[0128] When present as a separate formulation, each formulation contains, in addition to the active ingredient, a pharmaceutically acceptable carrier.

[0129] The kit of the present invention can be used to treat patients who have undergone complete resection of mucosal melanoma or to prevent recurrence or distant metastasis of mucosal melanoma in such patients.

[0130] Abbreviations

[0131] Throughout the specification and embodiments of this invention, the following abbreviations are used:

[0132] BID one dose, twice daily.

[0133] CDR Complementary Determinant Region

[0134] DFS (Disease-Free Survival)

[0135] FR Frame Area

[0136] IgG Immunoglobulin G

[0137] IHC Immunohistochemistry

[0138] OR Overall Response

[0139] ORR (Objective Response Rate)

[0140] DCR (Disease Control Rate)

[0141] OS Total Lifetime

[0142] mOS (mean overall survival)

[0143] PD disease progression

[0144] PFS (Progression-Free Survival)

[0145] mPFS mean progression-free survival

[0146] PR Partial Response

[0147] CR complete response

[0148] SD disease stable

[0149] DLT dose-limiting toxicity

[0150] MTD Maximum Tolerable Dose

[0151] Q2W, one dose every two weeks.

[0152] QD One dose per day

[0153] CSD Long-term Sunlight Type

[0154] non-CSD (non-long-term sunlight type)

[0155] IRC Independent Review Committee

[0156] AE adverse events

[0157] Adverse events occurring during TEAE / TEAEs treatment

[0158] Treatment-related adverse reactions of TRAEs / TRAEs

[0159] SAE serious adverse reactions

[0160] RO receptor occupancy rate

[0161] RECIST criteria for evaluating the efficacy of treatment in solid tumors

[0162] irRECIST criteria for evaluating the efficacy of treatment in immune-related solid tumors

[0163] DOR Relief Duration

[0164] Time to Remission (TTR)

[0165] BIRC Blinded Individual Review Committee

[0166] NE cannot be evaluated

[0167] RFS (Relapse-Free Survival)

[0168] DMFS has no distant metastasis survival.

[0169] IVGTT (intravenous infusion)

[0170] ECOG (Eastern Cooperative Oncology Group)

[0171] HDI High-Dose Interferon

[0172] HR risk ratio

[0173] PD-1 programmed death-1

[0174] PD-L1 programmed death-ligand-1

[0175] CPS combined positive score

[0176] The present invention is further illustrated by the following embodiments, but these embodiments should not be construed as limiting the invention. All references cited throughout this application are expressly incorporated herein by reference.

[0177] Example

[0178] Example 1: Clinical study of anti-PD-1 antibody as adjuvant therapy for mucosal melanoma

[0179] This study is a multicenter, open-label, randomized, parallel-controlled trial comparing the efficacy and safety of recombinant humanized anti-PD-1 monoclonal antibody (Toripalimab / JS001) injection or high-dose interferon (HDI) in patients with mucosal melanoma, with or without localized lymphoid disease, after complete resection, and assessing the population for optimal biomarker prediction. The clinical trial registration number is NCT03178123 (ClinicalTrials.gov).

[0180] 1.1 Study population

[0181] Main inclusion criteria: Eligible subjects must be (1) aged between 18 and 75 years; (2) pathologically confirmed as mucosal melanoma; (3) complete resection of the primary lesion with negative surgical margins; complete systemic staging examination before enrollment to confirm no regional or distant metastasis; (4) not receiving regular adjuvant therapy; (5) ECOG score of 0 or 1; (6) no contraindications to treatment and adequate organ and bone marrow function; enrolled for treatment within 4 months after surgery.

[0182] The main exclusion criteria are: 1) coexistence of other melanoma subtypes; 2) history of autoimmune diseases; 3) non-melanoma-like cancer within the past 5 years; 4) persistent infection; 5) patients with autoimmune diseases who have received systemic treatment within the past two years.

[0183] 6) Patients with melanoma resection who have received systemic adjuvant therapy; 7) Patients who have previously received anti-PD-1, anti-PD-L1, or anti-PD-L2 immunotherapy.

[0184] 1.2 Clinical Design

[0185] The study planned to enroll 148 patients, who were randomly assigned 1:1 to either the toripalimab group (experimental group) or the interferon group (control group). Randomization stratification factors were disease stage (I vs II vs III), primary site location (head and neck, non-head and neck), and PD-L1 expression status (positive, negative). Disease stage was defined according to the criteria in two previous studies (Lian B et al., Ann Oncol 2017 28:868-873; Cui C et al., Ann Surg Oncol 2018 25:2184-2192): T1, invasion of the mucosa or submucosa; T2, invasion of the muscularis propria; T3, invasion of the adventitia; T4, invasion of adjacent structures; N0, no regional metastatic nodes; N1, one or more regional metastatic lymph nodes; Stage I, T1–2N0; Stage II, T3–4 / xN0; Stage III, T1–4 / xN1.

[0186] Patients with head and neck mucosal melanoma (primarily nasopharynx and oropharynx) were randomly assigned to either the toripalimab group or the HDI group. Patients in the experimental group received toripalimab infusion (3 mg / kg IV drip every 2 weeks) for up to 1 year (27 treatments). Patients in the control group received interferon alpha-2b, with some patients eligible for low-dose escalation therapy depending on tolerance (poor tolerance: 3 MIU / m²). 2 / d1、6MIU / m 2 / d2、9MIU / m 2 / d3, well tolerated: 9 MIU d1 or directly 15 MIU / m 2 / d). Subsequent acceptance of 15 MIU / m 2 / d, days 1-5 of the week, for 4 weeks; then 9 MIU / m 2 / d, 3 times a week, for 48 weeks. For patients with head and neck mucosal melanoma, adjuvant radiotherapy should be initiated 6-8 weeks after starting recombinant humanized anti-PD-1 monoclonal antibody injection or high-dose interferon treatment, with a reference dose of CTVTB 60-64 Gy / 30 fractions, on days 1-5 of the week, for a total of 6 weeks.

[0187] Patients with non-head and neck mucosal melanoma were randomly assigned to either the toripalimab group or the HDI group. Patients in the experimental group received toripalimab infusion (3 mg / kg IV drip every 2 weeks) for a maximum of 1 year (27 treatments). Patients in the control group received interferon alpha-2b, and some patients may opt for low-dose escalation therapy depending on tolerance (poor tolerance: 3 MIU / m²). 2 / d1、6MIU / m 2 / d2、9MIU / m 2 / d3, well tolerated: 9MIU / m 2 / d or directly 15MIU / m 2 / d), subsequently accepting 15 MIU / m 2 / d, days 1-5 / week, for 4 weeks; then 9 MIU / d, 3 times a week, for 48 weeks.

[0188] Safety will be evaluated during treatment. The treatment study may be terminated if disease recurrence or metastasis occurs, intolerable toxicity occurs, informed consent is withdrawn, or other criteria for withdrawal from the study are met.

[0189] All enrolled participants will be followed up on overall survival outcomes (tumor re-evaluation frequency: every 12 weeks in the first year, every 16 weeks in the second year, every six months in years 3-5, and every year thereafter), and information on their subsequent anti-tumor treatment and survival will be collected. If a participant discontinues study treatment without documented evidence of disease recurrence or metastasis, their tumor will continue to be evaluated at the same frequency until tumor recurrence or metastasis or the participant begins new anti-tumor treatment, whichever occurs first. The primary endpoint is RFS in the intention-to-treat population. Secondary endpoints include DMFS, 2-year RFS rate, OS, and safety.

[0190] 1.3 Number of patients

[0191] From July 2017 to May 2019, 187 patients underwent screening, and 145 patients were randomly assigned 1:1 to either the toripalimab group (n=73, experimental group) or the interferon group (n=72, control group). The demographic statistics of the enrolled subjects are shown in Table 1.

[0192] Table 1: Demographic and baseline clinical characteristics of the study subjects

[0193]

[0194]

[0195] PD-L1 positive: defined as tumor cells with CPS ≥ 1% stained with JS311 IHC.

[0196] HDI, High-dose interferon-α2b; IQR, Interquartile range; T, Tumor stage; N, Nodal stage; PD-L1, Programmed death-ligand 1; CLND, Complete lymph node anatomy; LDH, Lactate dehydrogenase; ULN, Upper limit of normal.

[0197] 1.4 Test Drug

[0198] Investigational drug: Recombinant humanized anti-PD1 monoclonal antibody injection (JS001, generic name: toripalimab injection, trade name: Tuoyi) was provided free of charge by Shanghai Junshi Biosciences Co., Ltd. / Suzhou Zhonghe Biopharmaceutical Technology Co., Ltd.; specification: 240mg / 6ml / vial.

[0199] Control drug: Interferon α-2b injection was provided by Merck & Co. (Ganlen), 18 MIU / 1.2 ml / vial.

[0200] 1.5 Disease Assessment

[0201] Disease assessments are performed every 12 weeks in the first year, every 16 weeks in the second year, then every 6 months until years 3-5, and then annually according to RECIST 1.1. Patients with nasopharyngeal or oropharyngeal melanoma require additional nasopharyngeal or oropharyngeal CT or MRI. Any suspicious lymph node metastasis detected by superficial lymph node ultrasound should be histologically verified by lymph node resection or anatomy, if necessary.

[0202] 1.6 Efficacy and safety assessment

[0203] The efficacy analysis was performed on all randomly assigned patients. The primary endpoint, RFS, was defined as the time from randomization to any documented tumor recurrence, metastasis, or death.

[0204] Safety assessments were performed on all patients who had received at least one dose of the study treatment. Adverse events were assessed and recorded in accordance with the National Cancer Institute Common Terminology Criteria, version 4.0.

[0205] 1.7 PD-L1 Expression Analysis in Tumor Biopsies

[0206] Tumor PD-L1 IHC staining was performed in the central laboratory using JS311 antibody (Wang Z, Ying J, Xu J, et al. Safety, Antitumor Activity, and Pharmacokinetics of Toripalimab, a Programmed Cell Death 1 Inhibitor, in Patients with Advanced Non-Small Cell Lung Cancer: A Phase 1 Trial. JAMA Netw Open 2020 3:e2013770). PD-L1 positivity was defined as a proportion of tumor cells and / or infiltrating immune cells (CPS) ≥ 1%.

[0207] 1.8 Statistical Analysis

[0208] Validity:

[0209] For the primary efficacy endpoint, recurrence-free survival (RFS), it refers to the time from the date of randomization to any recorded tumor recurrence, metastasis, or death.

[0210] Survival times will be calculated using the non-parametric Kaplan-Meier method; if the impact of stratification factors such as disease stage on survival curves is to be studied, a stratified Kaplan-Meier method can also be used for analysis. The comparison of the two groups of survival curves will be performed using a two-tailed Log-rank test.

[0211] Patients for whom no events (such as recurrence) were observed were censored, and their survival time was the time between the start of the study and the last follow-up.

[0212] In addition, Cox proportional hazards regression was used to conduct multivariate analysis to explore various research factors that may affect the relapse-free survival curve.

[0213] For the OS endpoint, a Kaplan-Meier (KM) curve and median estimate will be provided, depending on the circumstances.

[0214] Security:

[0215] Safety and tolerability will be assessed through clinical review of all relevant parameters, including adverse events (AEs), laboratory tests, and vital signs.

[0216] Summary statistics (number, percentage, median, standard deviation, etc.) of safety endpoints will be provided as appropriate. For any adverse events (AEs), any serious AEs, any grade 3-5 AEs, any drug-related AEs, any serious drug-related AEs, any grade 3-5 drug-related AEs, drug discontinuation due to AEs, drug discontinuation due to AEs, any immune-related AEs (irAEs), and the incidence of specific AEs, summary statistics of number, percentage, and 95% confidence intervals will be provided.

[0217] A summary statistical analysis of the mean and standard deviation of laboratory test and vital sign changes relative to baseline will be provided for each treatment cycle. A summary statistical analysis of the number and percentage of laboratory values ​​that deteriorated relative to baseline will be provided according to the CTCAE grade.

[0218] 2. Research Results

[0219] From July 2017 to May 2019, 187 patients underwent screening, and 145 patients were randomly assigned 1:1 to either the toripalimab group (n=73, experimental group) or the interferon group (n=72, control group). The mean age of the patients was 58 years; most patients were female (62.8%); 80.7% had focal disease, 19.3% had focal lymph node disease, 37.2% had local excision ± CLND, and 62.8% had wide excision ± CLND; 51.0% were PD-L1 positive (CPS≥1,22C3), and 49.0% were PD-L1 negative. There were no differences in baseline characteristics between the two groups.

[0220] Of the 57 (39.3%) patients with primary head and neck tumors, 24 (85.7%) of the 28 patients in the HDI group and 26 (89.7%) of the 29 patients in the toripalimab group received adjuvant radiotherapy. A total of 52 patients completed the 1-year treatment, including 30 (41.1%) patients in the toripalimab group and 22 (30.6%) patients in the HDI group. The current study workflow is described in [link to study details]. Figure 5 .

[0221] As of March 31, 2021, the median follow-up time was 26.4 months in the HDI group and 25.5 months in the toripalimab group. As the first clinical study comparing immunotherapy with HDI for mucosal melanoma, this study showed that toripalimab and HDI, as adjuvant therapies, had similar remission survival rates (RFS) in patients with completely resected mucosal melanoma. Toripalimab not only significantly prolonged the RFS in the PD-L1-positive subgroup, but also demonstrated significantly better safety and tolerability than the HDI group, with a lower incidence of ≥ grade 3 treatment-related adverse events (TRAEs), meeting the pre-specified superiority margin of the primary endpoint.

[0222] The results of the primary efficacy endpoint and all secondary efficacy endpoints are as follows:

[0223] 2.1 Study on anti-tumor effects

[0224] 2.1.1 RFS, DMFS, OS values

[0225] As of March 31, 2021, a total of 97 relapse-free survival (RFS) events occurred among all 145 patients, with 46 RFS events (63.9%) in the HDI group and 51 RFS events (69.9%) in the toripalimab group. The median RFS values ​​were similar between the HDI and toripalimab groups, with 13.9 months (95% CI: 8.28–19.61) in the HDI group and 13.6 months (95% CI: 8.31–19.02) in the toripalimab group. The relapse hazard ratio (HR) was 1.05 (95% CI: 0.69–1.61; stratified p = 0.812). Figure 1 (See Table 2). The 1-year RFS rates in the HDI group and the toripalimab group were 52% (95% CI: 38.61–63.76) and 52.9% (95% CI: 40.33–63.90), respectively, and the 2-year RFS rates were 25.1% (95% CI: 14.75–36.83) and 30.5% (95% CI: 19.73–41.89), respectively. The relapse hazard ratio (HR) for each subgroup, i.e., the ratio of RFS events between the experimental group and the control group, is shown in Table 2. Figure 2 As stated above.

[0226] Table 2: Median RFS, DMFS, and OS data of the participants (as of March 31, 2021)

[0227]

[0228]

[0229] PD-L1+ is defined as tumor cells with CPS ≥ 1% stained by JS311 IHC.

[0230] NR indicates that it cannot be evaluated; NR: Not met;

[0231] After disease recurrence or metastasis, the rates of subsequent surgical treatment were similar in both groups. However, after treatment cessation, more patients in the HDI group received immunotherapy than in the toripalimab group (32.6% vs. 26.0%) (Table 3).

[0232] Table 3: Proportion of patients receiving subsequent immunotherapy

[0233]

[0234] HDI, high-dose interferon-α2b; PD-1, programmed cell death-1; CTLA4, cytotoxic T-lymphocyte-associated protein 4.

[0235] A total of 93 DMFS events occurred, including 44 in the HDI group (61.1%) and 49 in the toripalimab group (67.1%). The median DMFS values ​​for the HDI group and the toripalimab group were 14.6 months (95% CI: 8.34–21.26) and 16.3 months (95% CI: 10.94–21.09), respectively, with a hazard ratio of 1.00 for distant metastasis (95% CI: 0.65–1.54; stratified p = 0.994). The 1-year DMFS rates for the HDI group and the toripalimab group were 54.8% (95% CI: 41.25–66.50) and 58.2% (95% CI: 45.41–68.96), respectively, and the 2-year DMFS rates were 27.1% (95% CI: 16.20–39.18) and 32.0% (95% CI: 20.86–43.67), respectively.

[0236] The median overall survival (OS) was not reached in the HDI group (95% CI: 28.29–NR), while the median OS in the toripalimab group was 35.1 months (95% CI: 27.93–NR) (Table 2). The relative hazard ratio (HR) for mortality was 1.11 (95% CI: 0.66–1.84). HRs for OS in each subgroup are shown below. Figure 3 .

[0237] 2.1.2 Subgroup efficacy

[0238] 1) PD-L1 expression positive / negative subgroups

[0239] As of March 31, 2021, PD-1-positive tumors were found in 36 patients (50.0%) in the HDI group and 38 patients (52.1%) in the toripalimab group. In PD-L1-positive patients, the median remission duration (RFS) was 11.1 months (95% CI: 6.60–21.29) in the HDI group and 17.4 months (95% CI: 8.15–22.47) in the toripalimab group, with a hazard ratio (HR) of 1.00 (95% CI: 0.57–1.74; p = 0.988). Toripalimab significantly prolonged the RFS in the PD-L1-positive subgroup (Table 2 and...). Figure 4A ).

[0240] In patients with PD-L1-negative tumors, the median RFS was 14.6 months in the HDI group (95% CI: 5.72–21.29) and 11.3 months in the toripalimab group (95% CI: 8.21–18.00) (Table 2 and ). Figure 4BThe hazard ratio (HR) was 1.00 (95% CI: 0.56–1.77; p = 0.99) (Table 2 and...). Figure 4B ).

[0241] 2) Other subgroups

[0242] For patients with disease stages I, II, and III, the median RFS in the HDI group was 8.3 months (95% CI: 5.52–21.29), 14.0 months (95% CI: 5.65–21.26), and 21.7 months (95% CI: 10.25–26.58), respectively; while the median RFS in the toripalimab group was 16.3 months (95% CI: 8.25–30.62), 17.4 months (95% CI: 5.39–NR), and 13.6 months (95% CI: 2.60–17.31), respectively. Furthermore, sex, age, primary tumor location, or other baseline characteristics did not lead to significant differences in RFS.

[0243] 2.2 Safety and Tolerability Studies

[0244] As of March 31, 2021, the number of treatment-related adverse events (TEAEs) was 90.4% in the toripalimab group and 100% in the HDI group. Most TEAEs in the toripalimab group were grade 1 or 2. No treatment-related deaths or infusion reactions occurred. Grade ≥3 TEAEs occurred in 20 patients (27.4%) in the toripalimab group and 63 patients (87.5%) in the HDI group; Grade ≥3 TRAEs were reported in 8 patients (11%) in the toripalimab group and 61 patients (84.7%) in the HDI group (Table 4). Therefore, toripalimab has better safety and tolerability compared to the HDI group, with a lower incidence of Grade ≥3 treatment-related adverse events (TRAEs).

[0245] Table 4: Adverse events were reported in at least 15% of patients in both trials during treatment. #

[0246]

[0247]

[0248]

[0249] “ # "Common treatment-related adverse events" are defined as those occurring in ≥20% of patients in the toripalimab group.

[0250] Overall, in this first-ever randomized phase II trial comparing toripalimab and hemolytic angioplasty (HDI) in patients with completely resected mucosal melanoma, both therapies showed numerically superior median remission survival (RFS) compared to resection alone. Toripalimab significantly prolonged RFS in the PD-L1-positive subgroup. These findings highlight the potential utility of these two interventions as adjuvant therapy for mucosal melanoma. Furthermore, toripalimab demonstrated superior safety and tolerability compared to HDI, suggesting it is a better treatment option. sequence list <110> Shanghai Junshi Biosciences Co., Ltd. <120> Medications and methods for treating patients with completely resected mucosal melanoma. <130> 213607Z11CNCN <150> 202110532489.7 <151> 2021-05-17 <160> 10 <170> PatentIn version 3.5 <210> 1 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> LCDR1 <400> 1 Arg Ser Ser Gln Ser Ile Val His Ser Asn Gly Asn Thr Tyr Leu Glu 1 5 10 15 <210> 2 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> LCDR2 <400> 2 Lys Val Ser Asn Arg Phe Ser 1 5 <210> 3 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> LCDR3 <400> 3 Phe Gln Gly Ser His Val Pro Leu Thr 1 5 <210> 4 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> HCDR1 <400> 4 Asp Tyr Glu Met His 1 5 <210> 5 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> HCDR2 <400> 5 Val Ile Glu Ser Glu Thr Gly Gly Thr Ala Tyr Asn Gln Lys Phe Lys 1 5 10 15 Gly <210> 6 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> HCDR3 <400> 6 Glu Gly Ile Thr Thr Val Ala Thr Thr Tyr Tyr Trp Tyr Phe Asp Val 1 5 10 15 <210> 7 <211> 112 <212> PRT <213> Artificial Sequence <220> <223> VL <400> 7 Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly 1 5 10 15 Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 8 <211> 125 <212> PRT <213> Artificial Sequence <220> <223> VH <400> 8 Gln Gly Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Ala Pro Ile His Gly Leu Glu Trp Ile 35 40 45 Gly Val Ile Glu Ser Glu Thr Gly Gly Thr Ala Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Gly Ile Thr Thr Val Ala Thr Thr Tyr Tyr Trp Tyr Phe 100 105 110 Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 125 <210> 9 <211> 219 <212> PRT <213> Artificial Sequence <220> <223> LC <400> 9 Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly 1 5 10 15 Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu 115 120 125 Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe 130 135 140 Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln 145 150 155 160 Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser 165 170 175 Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu 180 185 190 Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser 195 200 205 Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210 215 <210> 10 <211> 452 <212> PRT <213> Artificial Sequence <220> <223> HC <400> 10 Gln Gly Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Glu Met His Trp Val Arg Gln Ala Pro Ile His Gly Leu Glu Trp Ile 35 40 45 Gly Val Ile Glu Ser Glu Thr Gly Gly Thr Ala Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Gly Ile Thr Thr Val Ala Thr Thr Tyr Tyr Trp Tyr Phe 100 105 110 Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr 115 120 125 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser 130 135 140 Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 145 150 155 160 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 165 170 175 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 180 185 190 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys 195 200 205 Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu 210 215 220 Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val 355 360 365 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Leu Gly Lys 450

Claims

1. Use of an anti-PD-1 antibody or an antigen-binding fragment thereof in the manufacture of a medicament or kit for adjuvant treatment of a patient with completely resected mucosal melanoma or preventing recurrence or distant metastasis of mucosal melanoma in the patient; wherein the anti-PD-1 antibody or an antigen-binding fragment thereof comprises light chain complementarity determining regions of amino acid sequences set forth in SEQ ID NOs: 1, 2, and 3, and heavy chain complementarity determining regions of amino acid sequences set forth in SEQ ID NOs: 4, 5, and 6.

2. Use according to claim 1, characterized in that, The mucosal melanoma is a head and neck mucosal melanoma or a non-head and neck mucosal melanoma.

3. Use according to claim 1, characterized in that, The mucosal melanoma is a mucosal melanoma that is positive for PD-L1 expression in an immunohistochemical staining analysis of a tumor tissue section.

4. The use according to claim 1, characterized in that, The mucosal melanoma is a head and neck mucosal melanoma, and the use further comprises administering one or more therapies to the individual in need thereof.

5. Use according to claim 4, characterized in that, The therapy is a chemotherapeutic agent, a biologic therapeutic agent, an immunogenic agent, an immunostimulatory cytokine, a cytokine-encoding immunostimulatory agent, or a radiation therapy.

6. Use according to any one of claims 1 to 5, characterized in that, The anti-PD-1 antibody or an antigen-binding fragment thereof comprises a light chain variable region of an amino acid sequence set forth in SEQ ID NO: 7, and a heavy chain variable region of an amino acid sequence set forth in SEQ ID NO:

8.

7. Use according to any one of claims 1 to 5, characterized in that, The anti-PD-1 antibody comprises a light chain of an amino acid sequence set forth in SEQ ID NO: 9, and a heavy chain of an amino acid sequence set forth in SEQ ID NO:

10.

8. Use according to any one of claims 1 to 5, characterized in that, The anti-PD-1 antibody is toripalimab.

9. Use according to any one of claims 1 to 5, characterized in that, The anti-PD-1 antibody or an antigen-binding fragment thereof is administered at a dose of 0.1 mg / kg to 10.0 mg / kg body weight of the individual, or a fixed dose selected from the group consisting of 120 mg, 240 mg, 360 mg, and 480 mg.

10. Use according to claim 9, characterized in that, The anti-PD-1 antibody or an antigen-binding fragment thereof is administered at a dose of 0.1 mg / kg, 0.3 mg / kg, 1 mg / kg, 2 mg / kg, 3 mg / kg, 5 mg / kg, or 10 mg / kg body weight of the individual, or a fixed dose selected from the group consisting of 120 mg, 240 mg, 360 mg, and 480 mg.

11. The use according to claim 9, characterized in that, The anti-PD-1 antibody or an antigen-binding fragment thereof is administered at a frequency of once every week, once every two weeks, once every three weeks, once every four weeks, or once a month.

12. The use according to claim 9, characterized in that, The anti-PD-1 antibody or an antigen-binding fragment thereof is administered at a dose of 1 mg / kg, 3 mg / kg, or 10 mg / kg body weight of the individual, or a fixed dose of 240 mg or 480 mg, once every two or three weeks.

13. The use according to claim 9, characterized in that, The anti-PD-1 antibody or an antigen-binding fragment thereof is administered in a liquid dosage form via a parenteral route.

14. The use according to claim 13, characterized in that, The liquid dosage form is an injection.

15. The use according to claim 13, characterized in that, The parenteral route is intravenous infusion.

16. The use according to claim 9, characterized in that, The anti-PD-1 antibody or an antigen-binding fragment thereof is administered in a cycle of one week, two weeks, three weeks, one month, two months, three months, four months, five months, half a year, or one year.

17. The use according to claim 16, characterized in that, The time of each administration cycle is the same or different, and the interval between each administration cycle is the same or different.

18. The use according to claim 5, characterized in that, The radiation therapy dose is CTV TB 60~64 Gy / 30 fractions administered on days 1-5 of each week for a total of 6 weeks.

19. The use according to claim 18, characterized in that, The anti-PD-1 antibody or an antigen-binding fragment thereof is administered simultaneously with a radiation therapy within 6-8 weeks of administration of the anti-PD-1 antibody or an antigen-binding fragment thereof.

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