CN115364208A - Medicine and method for treating patient with completely resected mucosal melanoma - Google Patents

Abstract

The invention relates to a medicine and a method for treating a patient with completely resected mucosal melanoma, in particular to application of an anti-PD-1 antibody or an antigen-binding fragment thereof in preparation of a medicine for treating the patient with completely resected mucosal melanoma or preventing recurrence or distant metastasis of the patient with the mucosal melanoma. The invention also relates to a kit for treating a patient who has had a mucosal melanoma completely resected or for preventing recurrence or distant metastasis of the mucosal melanoma in the patient. The medicine and the method can obviously prolong the RFS of a PDL1 expression positive patient and have better tolerance.

Description

Drugs and methods for treating patients with completely resected mucosal melanoma

technical field

The present invention relates to medicines and methods for treating patients with completely resected mucosal melanoma, in particular to anti-PD-1 antibodies or antigen-binding fragments thereof used in the preparation and treatment of patients with completely resected mucosal melanoma or to prevent recurrence or distant metastasis of the patient's mucosal melanoma use in medicines.

Background technique

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), but in Asia, mucosal melanoma ranks second among all melanoma types, accounting for about 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). Mucosal melanoma has a poorer prognosis than cutaneous melanoma. Previous studies have shown that the 5-year overall survival (OS) rate of cutaneous melanoma is about 50-80%, but that of mucosal melanoma 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 208 O 2184-2192). Moreover, early mucosal melanoma is prone to relapse after resection. A previous study showed that the median RFS of 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.

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

Previous clinical trials have shown that high-dose interferon (HDI) (IFN)-α2b can prolong relapse-free survival (RFS) and/or OS in patients with high-risk cutaneous melanoma (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, etal. High-dose interferon alfa-2b significantly prolongs relapse-free and overall survival compared with the GM2-KLH/QS-21 vaccine in patients withresected stage 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 mAbs have also shown them as adjuvant therapy in the treatment of resected high-risk cutaneous melanoma (resected high-risk cutaneous melanoma). melanomas) (Eggermont AMM, BlankCU, Mandala M, et al. Adjuvant Pembrolizumab versus Placebo in Resected StageIII 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 IVMelanoma. N Engl J Med 2017 377: 1824-1835). Although high-dose interferon is currently the only treatment approved for the adjuvant treatment of melanoma, it is not universally accepted as the standard of care. Mounting evidence surrounding a questionable survival benefit, high incidence of serious toxicity, and negligible benefit for patients with more disease makes it an unattractive option for most patients and clinicians (Schuchter L. Adjuvant Interferon Therapy for Melanoma: High-Dose, Low-Dose, NoDose, WhichDose J Clin Oncol 2004; 22:7-10). Therefore, there is a need for more effective treatments with an acceptable safety profile in adjuvant therapy.

Contents of the invention

The present invention provides the use of anti-PD-1 antibody or antigen-binding fragment thereof in the preparation of a drug for treating a patient with completely resected mucosal melanoma or preventing the recurrence or distant metastasis of the patient's mucosal melanoma.

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

In yet another aspect, the present invention provides an anti-PD-1 antibody or an antigen-binding fragment thereof, which is used for adjuvant treatment of a patient with completely resected mucosal melanoma or to prevent recurrence or distant metastasis of the patient's mucosal melanoma.

In one or more embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof of the present invention is any antibody that can specifically bind to PD-1 and can block the binding of PD-1 to its ligand PD-L1 or antigen-binding fragments.

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

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

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 with an amino acid sequence such as SEQ ID NO: 1, 2 and 3, and an amino acid sequence such as SEQ ID NO: 1, 2 and 3 ID NO: 4, 5 and 6 heavy chain complementarity determining region.

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 an amino acid sequence such as SEQ ID NO: 8 Heavy chain variable region indicated.

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

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.

In one or more embodiments, the mucosal melanoma described in the present invention is head and neck mucosal melanoma, and the use further comprises the combined administration of one or more therapies to individuals in need; preferably, the therapies are Radiation Therapy.

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 about 120 mg, 240 mg, 360 mg or 480 mg fixed dose, preferably about 3 mg/kg body weight of the subject or about 240 mg fixed dose.

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 two weeks.

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 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, 480 mg fixed dose administered every two or three weeks.

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 3 mg/kg individual body weight, or a fixed dose of 240 mg, administered once every two weeks.

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 injection, via parenteral route, such as intravenous infusion.

In one or more embodiments, the administration cycle of the anti-PD-1 antibody or antigen-binding fragment thereof of 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 more, optionally, the time of each administration cycle is the same or different, and the interval between each administration cycle is the same or different.

In one or more embodiments, the radiotherapy dose of the present invention is CTV _TB 60-64Gy/30 times, administered on the 1st-5th day of each week, for a total of 6 weeks; preferably, anti-PD-1 antibody or Antigen-binding fragments thereof are administered within 6-8 weeks concurrently with radiation therapy.

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

In one or more embodiments, the additional therapy of the invention is selected from a chemotherapeutic agent, a biotherapeutic agent, an immunogenic agent, an immunostimulatory cytokine, an encoded immunostimulatory cytokine, or radiation therapy.

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

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

In one or more embodiments, the method of the present invention has fewer tertiary and above treatment-related adverse effects (TEAEs) than high-dose interferon alpha 2b (HDI) adjuvant therapy. Preferably, Grade III and above treatment-related adverse effects (TEAEs) are about 10%-60%, 10%-50%, 10%-40%, 10%-30%, or 10%-20% lower than HDI.

In one or more embodiments, the adjuvant therapy method described in the present invention has a better effect than the adjuvant therapy method of high-dose interferon α-2b (HDI). Preferably, the relapse-free survival (RFS) of PD-L1 positive patients is about 50%, 40%, 30%, 20% or 10% higher than HDI.

In one or more embodiments, the adjuvant therapy method described in the present invention has a better effect than the adjuvant therapy method of high-dose interferon α-2b (HDI). Preferably, the relapse-free survival (RFS) of PD-L1 positive patients is about 10-50%, 10-40%, 10-30%, or 10-20% higher than HDI.

In yet another aspect, the present invention provides a kit comprising one or more single pharmaceutical dosage 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 An antigen-binding fragment thereof is as described in any one herein. The kit of the present invention can be used to treat a patient with completely resected mucosal melanoma or prevent the recurrence or distant metastasis of the patient's mucosal melanoma.

In yet another aspect, the present invention provides a method of using biomarkers to predict the efficacy of an anti-PD-1 antibody or an antigen-binding fragment thereof in treating a patient with completely resected mucosal melanoma to prevent recurrence or distant metastasis of the patient's mucosal melanoma.

Description of drawings

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

Figure 2: Hazard ratios (HR) for recurrence-free survival (RFS) in each subgroup.

Figure 3: Hazard ratios (HR) for overall survival (OS) in each subgroup.

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

Figure 5: Flowchart of the current study.

Detailed ways

The present invention relates to a treatment method for complete resection of mucosal melanoma. The methods of the invention comprise administering an anti-PD-1 antibody or antigen-binding fragment thereof described herein to a patient in need thereof.

the term

In order to make the present invention easier to understand, certain technical terms are specifically defined below. Unless otherwise specified herein, all technical and technical terms used herein have the meanings commonly understood by those of ordinary skill in the art to which the present invention belongs.

"Administering", "administering" 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. The route of administration of the anti-PD-1 antibody includes intravenous, intramuscular, subcutaneous, peritoneal, spinal or other parenteral routes of administration, such as injection or infusion. "Parenteral administration" means administration other than enteral or topical administration, usually by injection, including but not limited to intravenous, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intrasaccular , Intratracheal, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, intra-articular, subcapsular, subarachnoid, intraspinal, intradural and intrasternal injection and infusion and in vivo electroporation.

An "adverse effect" (AE) as described herein is any unfavorable and often unintentional or undesired sign, symptom or disease associated with the use of a medical treatment. For example, adverse reactions may be related to 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 be of the same or a different level of severity.

"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 various methods known in the art, such as using calipers after the tumor has been removed from the subject, or using imaging techniques (such as ultrasound, bone scan, computed tomography, etc.) (CT) or magnetic resonance imaging (MRI) scan) to measure its size.

The term "tumor size" refers to the overall size of the tumor, which can be measured as the length and width of the tumor. Tumor size can be determined by various methods known in the art, such as measuring the tumor after it has been removed from the subject using calipers, or while in vivo using imaging techniques such as bone scans, ultrasound, CT or MRI scans. size.

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

An "antibody" as used herein refers to any form of antibody that can achieve a desired biological activity or binding activity. Accordingly, 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 single domain antibodies of camelid origin. An "antibody" specifically binds 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 comprising three constant domains CH1, CH2 and CH3. Each light chain comprises a light chain variable region (VL) and a light chain constant region comprising a constant domain, CL. The VH and VL regions can be further subdivided into hypervariable regions called complementarity determining regions (CDRs), interspersed with more conserved regions called framework regions (FRs). In general, from N-terminus to C-terminus, both light and heavy chain variable domains comprise FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4. Amino acids are generally assigned to each domain according to the definitions in: 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.

The carboxy-terminal portion of the heavy chain defines a constant region primarily responsible for effector functions. Generally, human light chains are classified into kappa chains and lambda chains. Human heavy chains are typically classified as mu, delta, gamma, alpha, or epsilon, and define the antibody's 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, IgGl, IgG2, IgG and IgG4.

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; Non-human Abs can be humanized by recombinant methods to reduce their immunogenicity in humans.

Unless expressly stated otherwise, an "antibody fragment" or "antigen-binding fragment" as used herein refers to an antigen-binding fragment of an antibody, i.e., an antibody fragment that retains the ability of a full-length antibody to specifically bind to an antigen, such as retaining one or Fragments of multiple 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.

"Chimeric antibody" refers to an antibody, and fragments thereof, in which a portion of the heavy and/or light chain is identical or homologous to the corresponding sequence in an antibody derived from a specific species (e.g., human) or belonging to a specific antibody class or subclass , while the rest 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, as long as it exhibits the desired biological activity.

"Human antibody" refers to an antibody that comprises only human immunoglobulin sequences. If the human antibody is produced in a mouse, mouse cells, or hybridomas derived from mouse cells, it may contain murine carbohydrate chains. Similarly, a "mouse antibody" or "rat antibody" refers to an antibody comprising only mouse or rat immunoglobulin sequences, respectively.

"Humanized antibody" refers to a form of antibody that contains sequences from non-human (eg, murine) antibodies as well as human antibodies. Such antibodies contain minimal sequence derived from a single side of a non-human immunoglobulin. Typically, a humanized antibody will comprise substantially all of at least one, and usually two variable domains, in which all or substantially all hypervariable loops correspond to those of a non-human immunoglobulin, and in which all or substantially all hypervariable loops correspond to those of a non-human immunoglobulin, and all or substantially all The FR regions are those of human immunoglobulins. The humanized antibody optionally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin.

The term "immunotherapy" refers to the treatment of a subject suffering from a disease or at risk of infection or suffering relapse of the disease by methods including 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 administration of an active agent to a subject, for the purpose of reversing, alleviating, ameliorating, slowing down or preventing symptoms, complications or the onset, progression, severity or recurrence of a condition, or biochemical indicators associated with a disease.

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

A "therapeutically effective amount" or "therapeutically effective dose" of a drug or therapeutic agent is any amount of the drug that protects a subject from the onset of a disease or promotes regression of a disease when used alone or in combination with another therapeutic agent that Disease regression is evidenced by a decrease in the severity of disease symptoms, an increase in the frequency and duration of asymptomatic periods of disease, or prevention of impairment or disability resulting from disease affliction. The ability of a therapeutic agent to promote disease regression can be assessed 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 in vitro assays The activity of the drug was determined in the assay.

A therapeutically effective amount of a drug includes a "prophylactically effective amount", i.e. any amount that inhibits the development or recurrence of cancer when administered alone or in combination with an antineoplastic agent to a subject at risk of developing cancer or to a subject suffering from cancer recurrence medicine.

"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.

As used herein, unless expressly indicated otherwise, "CDR" refers to an immunoglobulin variable region that is a complementarity determining region defined using the Kabat numbering system.

"Therapeutic anti-PD-1 monoclonal antibody" refers to an antibody that specifically binds to a particular mature form of PD-1 expressed on the surface of certain mammalian cells. Mature PD-1 has no presecretion leader sequence, or leader peptide. The terms "PD-1" and "mature PD-1" are used interchangeably herein and are to be understood as the same molecule unless clearly defined otherwise, or clearly evident from the context.

As described herein, a therapeutic anti-human PD-1 antibody or anti-hPD-1 antibody refers to a monoclonal antibody that specifically binds to mature human PD-1.

"Framework region" or "FR" as used herein refers to the variable region of an immunoglobulin excluding the CDR regions.

"Isolated antibody or antigen-binding fragment thereof" refers to a purified state and in which case the specified molecule is substantially free of other biological molecules, such as nucleic acids, proteins, lipids, carbohydrates, or other material (such as cell debris or growth culture medium).

"Patient", "Patient" or "Subject" means any single subject, usually a mammal, including humans and other mammals, in need of a medical procedure or participating in a clinical trial, epidemiological study or used as a control, Such as horses, cows, dogs or cats.

"RECIST 1.1 Response Criteria" as used herein refers to the definition described by Eisenhauver et al., E.A. et al., Eur. J Cancer 45:228-247 (2009) for target or non-target damage based on the context of the measured response. Before immunotherapy, it was the most commonly used standard for evaluating response in solid tumors. However, with the advent of the era of immunity, there have been many problems that have not appeared in the evaluation of tumors before. Therefore, based on the emerging phenomenon caused by immunotherapy itself, in 2016, the RECIST working group reviewed the existing "RECIST v.1.1 "After the revision, a new judgment standard is proposed, namely the "irRECIST standard" described in this article, which aims to better evaluate the efficacy of immunotherapy drugs.

The term "ECOG" scoring standard is an indicator of the patient's general health and ability to tolerate treatment from the physical strength of the patient. ECOG physical status scoring standard score: 0 points, 1 point, 2 points, 3 points, 4 points and 5 points. A score of 0 means that the activity ability is completely normal, and there is no difference in the activity ability before the onset of the disease. A score of 1 refers to the ability to ambulate freely and engage in light physical activity, including general housework or office work, but not to engage in heavier physical activity.

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

A "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.

"Treating" cancer as described herein refers to a subject suffering from cancer or diagnosed with cancer using a treatment regimen as described herein (such as administering an anti-PD-1 antibody) to achieve at least one positive therapeutic effect (such as, cancer decreased cell number, decreased tumor volume, decreased rate of cancer cell infiltration into surrounding organs, or decreased rate of tumor metastasis or tumor growth). Positive treatment effects in cancer can be measured in various ways (see W.A. 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 value of treated tumor volume/median value of 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 CR or PR as well as the amount of time a patient experiences SD. DFS is the length of time a patient remains disease-free during and after treatment. OS refers to the increase in life expectancy compared to naive or untreated individuals or patients. The treatment regimen of the combinations of the invention effective in treating a cancer patient may vary depending on a variety of 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 the invention may not be effective in achieving a positive therapeutic effect in every subject, they should be effective and achieve a positive therapeutic effect in a statistically significant number of subjects.

The terms "administration mode" and "dosage regimen" are used interchangeably and refer to the dosage and time of use of each therapeutic agent in the combination of the present invention.

The term "immunohistochemistry (IHC)" refers to the use of the principle of specific binding of an antigen to an antibody to determine the antigen in tissue cells by chemically reacting the chromogenic agent (fluorescein, enzyme, metal ion, isotope) of the labeled antibody. (polypeptides and proteins), and methods for their localization, qualitative and relative quantitative research. In some embodiments of the present invention, before anti-PD-1 antibody treatment, PD-L1 detection is performed on the tumor tissue sample of the subject, and the detection uses Roche's anti-human PD-L1 antibody SP142 (Cat No: M4422 ) for staining experiments. In some embodiments, tumor cells are defined as PD-L1 positive by >1% membrane staining intensity.

As used herein, the terms "cancer" or "malignancy" refer to a broad variety of diseases characterized by the uncontrolled growth of abnormal cells in the body. Unregulated cell division, growth division, and growth lead to the formation of malignant tumors, which invade adjacent tissues and can also metastasize to distant parts of the body through the lymphatic system or bloodstream. Examples of cancers suitable for treatment or prevention using the methods, medicaments, and kits of the 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's lymphoma, non-Hodgkin's lymphoma, acute myelogenous leukemia, multiple myeloma , gastrointestinal (tract) 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.

Herein, the term "tumor mutational burden (TMB)" refers to the total number of detected somatic gene coding errors, base substitutions, gene insertion or deletion errors per million bases. In some embodiments of the invention, tumor mutational burden (TMB) is estimated by analysis of somatic mutations, including coding base substitutions and megabase insertions of the panel sequences studied.

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

anti-PD-1 antibody

Herein, "PD-1 antibody" refers to binding to the PD-1 receptor, blocking the combination of PD-L1 expressed on cancer cells and PD-1 expressed on immune cells (T, B, NK cells) and preferably Any chemical compound or biomolecule that blocks the binding of PD-L2 expressed on cancer cells to PD-1 expressed on immune cells. Alternatives 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 there are PDCD1L2, PDL2, B7-DC and CD273. In any of the methods of treatment, medicaments and uses of the 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 PD-L1. PD1 binding. The human PD-1 amino acid sequence can be found at NCBI locus number: NP_005009. The amino acid sequences of human PD-L1 and PD-L2 can be found in NCBI locus numbers: NP_054862 and NP_079515, respectively.

Herein, when an "anti-PD-1 antibody" is referred to, unless otherwise stated or described, the term includes antigen-binding fragments thereof.

The anti-PD-1 antibody suitable for any use, therapy, drug and kit described in the present invention binds to PD-1 with high specificity and high affinity, blocks the combination of PD-L1/2 and PD-1, and inhibits PD-1 signal transduction, so as to achieve the immunosuppressive effect. In any of the uses, therapies, medicaments, and kits disclosed herein, anti-PD-1 antibodies include full-length antibodies themselves, as well as antibodies that bind to the PD-1 receptor and behave like intact Abs in inhibiting ligand binding and upregulating the immune system. Antigen-binding portions or fragments of functional properties. In some embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof is an anti-PD-1 antibody or antigen-binding fragment thereof that cross-competes with toripalimab for binding to human PD-1. In other embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof is a chimeric, humanized or human Ab or antigen-binding fragment thereof. In certain embodiments for treating a human subject, the Ab is a humanized Ab.

In some embodiments, anti-PD-1 antibodies for any of the uses, therapies, medicaments and kits described herein include monoclonal antibodies (mAbs) or antigen-binding fragments thereof that specifically bind to PD-1, And preferably specifically bind to human PD-1. mAbs can be human, humanized, or chimeric antibodies, and can include human constant regions. 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 suitable for any use, therapy, drug and kit described in the present invention The antigen-binding fragment thereof comprises a heavy chain constant region of a human IgGl 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 the S228P mutation, which replaces the hinge region with a proline residue normally present at the corresponding position in antibodies of the IgG1 isotype serine residues in .

Preferably, in any embodiment of the uses, therapies, medicaments and kits of the present invention, the PD-1 antibody is a monoclonal antibody or an antigen-binding fragment thereof, and its light chain CDR is SEQ ID NO: 1, 2 The amino acids shown in and 3, the heavy chain CDRs are the amino acids shown in SEQ ID NO: 4, 5 and 6.

More preferably, in any embodiment of the uses, therapies, medicaments and kits of the present invention, the PD-1 antibody specifically binds to human PD-1 and comprises: (a) comprising SEQ ID NO: 7 and (b) a monoclonal antibody comprising the heavy chain variable region of SEQ ID NO:8.

Further preferably, in any embodiment of the uses, therapies, medicaments and kits of the present invention, the PD-1 antibody specifically binds to human PD-1 and comprises: (a) comprising SEQ ID NO: 9 and (b) a monoclonal antibody comprising the heavy chain of SEQ ID NO:10.

Table A provides the amino acid sequence numbering of the light chain CDR and heavy chain CDR of the exemplary anti-PD-1 antibody mAb used in the uses, therapies, medicaments and kits of the present invention, and the numbering is according to the Kabat rule:

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

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

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

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

In certain embodiments, the anti-PD-1 antibodies that can be used in any of the uses, therapies, medicaments, and kits described in the present invention also include antibodies that specifically bind to PD-L1 to block the combination of PD-L1 and PD-1. Anti-PD-L1 monoclonal antibodies, such as nivolumab, pembrolizumab, toripalimab, sintilimab, camrelizumab, tislelizumab, cemiplimab.

"PD-L1" expression or "PD-L2" expression as described herein 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 in IHC analysis of tumor tissue sections using a diagnostic PD-L antibody 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 a desired PD-L target, such as PD-L1 or PD-L2.

For methods of quantifying PD-L1 protein expression in IHC analysis of tumor tissue sections, see, but not limited to, Thompson, R.H. et al., PNAS 101(49): 17174-17179 (2004); Taube, J.M. et al., Sci TranslMed 4, 127ra37 (2012); and Toplian, S.L. et al., New Eng. J. Med. 366(26):2443-2454 (2012) et al.

One approach employs a simple binary endpoint of positive or negative PD-L1 expression, where a positive result is defined by the percentage of tumor cells showing histological evidence of cell surface membrane staining. The positive expression of PD-L1 was defined as the tumor tissue section counted as more than 1% of the total tumor cells.

In another approach, PD-L1 expression in tumor tissue sections is quantified in tumor cells as well as in infiltrating immune cells. The percentages of tumor cells and infiltrating immune cells exhibiting membrane staining were quantified separately from < 1%, 1% to 50%, and then 50% up to 100%. For tumor cells, if the score is ≤1%, the PD-L1 expression count is negative, and if the score is >1%, it is positive.

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

In some embodiments, PD-L1 positive is defined as CPS≥1% of tumor cells stained by JS311 IHC. CPS denotes Positive Combined Score. CPS evaluation criteria are: tumor cells with any intensity membrane staining, membrane/cytoplasmic staining lymphocytes/macrophages directly associated with tumor cells, the proportion of the above cells relative to tumor cells (at least one hundred), but should be excluded All necrotic cells, stromal cells, carcinoma in situ and other immune cells (including but not limited to neutrophils, eosinophils, plasma cells) were stained.

Complementary therapy

As used herein, "adjuvant therapy" or "adjuvant therapy" is understood as the administration of one or more drugs to a patient following surgical resection of one or more cancerous tumors in which all tumors have been removed from the patient. Detectable and resectable disease (e.g., cancer), but there is also a statistical risk of recurrence due to occult disease, where the goal is to reduce the likelihood or severity of recurrence or disease, or to delay the biological manifestations of disease recurrence Appear.

Effective amount and excision

The term "effective amount" refers to the amount of a drug or agent that elicits the biological or medical response sought by, for example, a researcher or clinician in a tissue, system, animal or human. Moreover, the term "therapeutically effective amount" refers to any amount that results in improved treatment, cure, prevention, or alleviation of a disease, disorder, or side effect, or a reduced rate of disease or disorder progression, compared to a corresponding subject not receiving that amount . The term also includes within its scope amounts effective to enhance normal physiological function. A therapeutically effective amount of a drug also includes a "prophylactically effective amount".

With respect to 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 otherwise undesired cellular proliferation. In some embodiments, an effective amount is an amount sufficient to prevent or delay tumor recurrence. An effective amount can be administered in one or more administrations. An effective amount of the drug or composition can: (i) reduce the number of cancer cells; (ii) reduce tumor size; (iii) inhibit, delay, slow down to a certain extent and prevent cancer cells from infiltrating into peripheral organs; (iv) ) Inhibition means slowing to a certain extent and can prevent tumor metastasis; (v) inhibiting tumor growth; (vi) preventing or delaying tumor occurrence and/or recurrence; and/or (vii) reducing one or more cancer-related Symptoms relieved to a certain extent.

The term "resection" refers to the surgical removal of malignant tissue characteristic of melanoma from a human patient. Resection is understood to mean the removal of malignant tissue such that the presence of remaining malignant tissue in said patient cannot be detected with existing methods. Resection is understood to mean the removal of melanoma such that the presence of remaining melanoma in said patient cannot be detected. Resection can include complete resection or partial resection.

recurrence and distant metastasis

The term "recurrence" refers to the occurrence of local recurrence of tumor or melanoma, and refers to the reappearance of tumor or melanoma within 2cm of the original tumor area in patients who have had resected/completely resected tumor or melanoma. Local recurrence of melanoma can occur at any site of the primary melanoma or in the closest regional lymph node to the primary melanoma.

The term "distant metastasis" refers to the distant metastasis of tumor or melanoma, which refers to patients who have resected/completely resected the tumor or melanoma, and the tumor or melanoma reappears beyond 2cm of 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, and others.

Combination Therapies, Dosage, and Administration Schedules

The combination therapies of the invention may also comprise one or more additional therapies. Additional therapeutic agents can be, for example, chemotherapeutics, biotherapeutics, 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 (eg, IL-2, IFN-onoma, GM-CSF), cells transfected with genes encoding immunostimulatory cytokines (such as but not limited to GM-CSF), or radiation therapy.

The choice of a dosing regimen (also referred to herein as an administration regimen) for a drug of the invention depends on several factors, including the subject's individual serum or tissue turnover rate, level of symptoms, overall immunogenicity, and target cell , the accessibility of tissues or organs. Preferably, the dosing regimen maximizes the amount of each therapeutic agent delivered to the patient, consistent with acceptable levels of side effects. Thus, the dosage and frequency of administration of each biotherapeutic and chemotherapeutic agent will depend in part on the particular therapeutic agent, the severity of the cancer being treated, and the characteristics of the patient. Guidance for selecting appropriate doses of antibodies, cytokines, and small molecules is available. Determination of an appropriate dosage regimen can be made by a clinician, e.g., by reference to parameters or factors in the art known or suspected to affect therapy or expected to affect therapy, and will depend, e.g., on the patient's clinical history (e.g., previous treatment ), the type and stage of cancer being treated, and biomarkers of response to one or more therapeutic agents in the combination therapy.

Each therapeutic agent or therapy of the combination therapy of the invention may 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 in any order Apply sequentially. The therapeutic agents in 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 on different dosing schedules (e.g., chemotherapeutics are administered at least daily and Sequential administration is especially useful when the biotherapeutic is administered less frequently (eg, once a week, every two weeks, or every three weeks).

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

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

Anti-PD-1 antibodies or antigen-binding fragments thereof of the invention can be administered by continuous infusion or by interval doses. Single administration doses may 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 of an individual, or as fixed doses from about 120 mg to about 480 mg. For example, the dose 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 kg, about 8 mg/kg, about 9 mg/kg, or about 10 mg/kg body weight of the individual. Alternatively, a fixed dose of about 120 mg, 240 mg, 360 mg or 480 mg may be administered. Dosage regimens are generally designed to achieve exposures that result in sustained receptor occupancy (RO) based on the typical pharmacokinetic properties of Abs. A representative dosing regimen may be about once weekly, about once every two weeks, about once every three weeks, about once every four weeks, about once a month, or longer. In some embodiments, the anti-PD-1 antibody is administered to the individual about once every two weeks.

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

In some embodiments, the mucosal melanoma is head and neck mucosal melanoma, within 6-8 weeks of administration of the anti-PD-1 antibody or antigen-binding fragment thereof, concurrently with radiation therapy. The dose of radiotherapy was CTVTB 60-64Gy/30 times, administered on the 1st-5th day of each week for a total of 6 weeks.

The anti-PD-1 antibody or antigen-binding fragment thereof of the present invention and the administration period of chemotherapy are one week, two weeks, three weeks, one month, two months, three months, four months, five months, half a year or more Over a long period of time, optionally, the timing of each dosing cycle can be the same or different, and the interval between each dosing cycle can be the same or different. In some embodiments, one dosing/treatment cycle is two weeks. In some embodiments, toripalimab is administered at about 3 mg/kg biweekly in a dosing cycle. In some embodiments, one dosing/treatment cycle of radiation therapy is one week. In some embodiments, in one administration cycle, the dose of radiotherapy is CTVTB 60-64Gy/30 times, administered on the 1st-5th day of each week. The administration time of the anti-PD-1 antibody or antigen-binding fragment thereof of the present invention is no more than 1 year. The administration time of the radiotherapy of the present invention is 6 weeks, and it is administered simultaneously within 6-8 weeks from the administration of the anti-PD-1 antibody or its antigen-binding fragment.

Treatments and uses

The purpose of the present invention is to treat patients with completely resected mucosal melanoma and prevent recurrence or distant metastasis of their mucosal melanoma. The present invention finds that complete resection can be effectively prevented by administering an effective amount of any anti-PD-1 antibody or antigen-binding fragment thereof described herein or a pharmaceutical composition containing the anti-PD-1 antibody or antigen-binding fragment thereof For patients with recurrent or distantly metastatic mucosal melanoma, the recurrence-free survival time RFS and/or distant metastasis-free survival period DMFS of patients can be improved; at the same time, the overall survival period of patients can be improved. In some embodiments, by administering an effective amount of any anti-PD-1 antibody or antigen-binding fragment thereof described herein or a pharmaceutical composition containing the anti-PD-1 antibody or antigen-binding fragment thereof, the patient's Medication safety and tolerability.

Therefore, the present invention provides the use of the aforementioned anti-PD-1 antibody or antigen-binding fragment thereof of the present invention in the preparation of a drug for preventing or treating a patient with completely resected mucosal melanoma or preventing recurrence or distant metastasis of the patient's mucosal melanoma.

The present invention also provides a method for preventing or treating a patient with completely resected mucosal melanoma or preventing recurrence or distant metastasis of the patient's mucosal melanoma, which comprises administering to the patient an effective amount of the anti-PD-1 antibody or its antigen of the present invention Combine fragments. The effective amount includes prophylactic effective amount and therapeutic effective amount.

The present invention also provides the aforementioned anti-PD-1 antibody of the present invention or an antigen-binding fragment thereof, which is used for treating a patient with completely resected mucosal melanoma or preventing recurrence or distant metastasis of the patient's mucosal melanoma.

The invention also provides for the co-administration of a therapeutically effective amount of one or more therapies (eg, treatment modalities and/or other therapeutic agents) to a subject. In some embodiments, the therapy includes surgery and/or radiation therapy.

In this article, "completely resected mucosal melanoma" refers to complete resection of the primary tumor with negative surgical margins.

In some embodiments, the methods or uses provided herein further comprise administering one or more therapies (e.g., treatment modalities and/or other therapeutic agents) to an individual in need thereof. Antibodies of the invention can be used alone or in combination with other therapeutic agents in therapy. For example, it can be co-administered with at least one additional therapeutic agent. In some embodiments, the methods or uses provided herein further comprise administering radiation therapy (radiotherapy) to the individual in need thereof.

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

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

Preferably, the method, use or anti-PD-1 antibody described in any embodiment of the present invention is especially suitable for mucosal melanoma with positive expression of PD-L1 in the immunohistochemical staining analysis of tumor tissue sections.

The preferred anti-PD-1 antibody for treating a patient with completely resected mucosal melanoma or preventing the recurrence or distant metastasis of the patient's mucosal melanoma can be as described in any embodiment herein, more preferably the light chain CDR is SEQ ID NO : amino acids shown in 1, 2 and 3, heavy chain CDRs are antibodies shown in SEQ ID NO: 4, 5 and 6 amino acids, more preferably comprising the light chain variable region shown in SEQ ID NO: 7 and SEQ ID The monoclonal antibody of the heavy chain variable region shown in NO: 8, more preferably a monoclonal antibody comprising the light chain shown in SEQ ID NO: 9 and the heavy chain shown in SEQ ID NO: 10, more preferably WO2014206107 The humanized antibodies described in 38, 39, 41 and 48, most preferably toripalimab.

In a particularly preferred embodiment, the present invention provides a method for treating a patient with completely resected mucosal melanoma or preventing recurrence or distant metastasis of mucosal melanoma in the patient, said method comprising administering to the patient a therapeutically or prophylactically effective amount of Trepro Limumab; preferably, the patient is positive for PD-L1 expression. In certain embodiments, the mucosal melanoma is a non-head and neck mucosal melanoma. In certain embodiments, the mucosal melanoma is head and neck mucosal melanoma, and the method further comprises concurrently administering radiation therapy within 6-8 weeks of administering toripalimab.

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 drug for treating a patient with completely resected mucosal melanoma or preventing recurrence or distant metastasis of the patient's mucosal melanoma. Preferably, the expression of PD-L1 is positive (CPS ≥ 1%) in the immunohistochemical staining analysis of the tumor tissue section of the patient with completely resected mucosal melanoma. In certain embodiments, the mucosal melanoma is a non-head and neck mucosal melanoma. In certain embodiments, the mucosal melanoma is a head and neck mucosal melanoma, and the use further includes concurrently administering radiation therapy within 6-8 weeks of administering toripalimab.

pill box

The invention also provides a kit comprising one or more single pharmaceutical dosage units of the anti-PD-1 antibody or antigen-binding fragment thereof according to any embodiment herein.

When present in the form of independent preparations, each preparation contains a pharmaceutically acceptable carrier in addition to the active ingredient.

The medicine kit of the present invention can be used for treating patients with completely resected mucosal melanoma or preventing the recurrence or distant metastasis of the patients' mucosal melanoma.

Acronym

Throughout the specification and examples of the present invention, the following abbreviations are used:

One dose BID, 2 times a day

CDR complementarity determining region

DFS disease-free survival

FR framework region

IgG Immunoglobulin G

IHC Immunohistochemistry

OR overall response

ORR objective response rate

DCR disease control rate

OS overall survival

mOS mean overall survival

PD disease progression

PFS progression-free survival

mPFS mean progression-free survival

PR partial answer

CR complete response

SD stable disease

DLT dose limiting toxicity

MTD maximum tolerated dose

Q2W One dose every two weeks

QD One dose per day

CSD long-term sunshine type

non-CSD non-long-term sunshine type

IRC Independent Review Committee

AE adverse event

Adverse events during TEAE/TEAEs treatment

TRAE/TRAEs treatment-related adverse reactions

SAE serious adverse reaction

RO receptor occupancy

RECIST Response Evaluation Criteria for Solid Tumors

irRECIST Response Evaluation Criteria for Immune-Related Solid Tumors

DOR duration of response

Time from TTR to disease remission

BIRC Blinded Individual Review Committee

NE cannot evaluate

RFS recurrence-free survival

DMFS distant metastasis-free survival

ivgtt intravenous drip

ECOG Eastern Cooperative Oncology Group

HDI high dose interferon

HR hazard ratio

PD-1 Programmed Death-1

PD-L1 Programmed Death Ligand-1

CPS Combined Positive Score

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

Example

Example 1: Clinical research of anti-PD-1 antibody in adjuvant therapy of mucosal melanoma

This study is a multi-center, open, randomized, parallel controlled study comparing completely resected mucosal melanoma patients with or without regional lymphatic disease receiving recombinant humanized anti-PD-1 monoclonal antibody (Terry Efficacy and safety of pulimumab/Toripalimab/JS001) injection or high-dose interferon (HDI) treatment, and the population for which best biomarker prediction was evaluated. The clinical trial registration number: NCT03178123 (ClinicalTrials.gov).

1.1 Study population

Main inclusion criteria: Eligible subjects must (1) be between 18 and 75 years old; (2) be diagnosed with mucosal melanoma confirmed by histopathology; (3) complete resection of the primary tumor, surgical resection Margin (-); complete systemic staging examination before enrollment confirmed no regional or distant metastasis; (4) did not receive formal adjuvant therapy; (5) ECOG score was 0 or 1; (6) had no treatment contraindications and had adequate organ and bone marrow function; enrolled within 4 months after surgery for treatment.

Main exclusion criteria: 1) other melanoma subtypes coexist; 2) autoimmune history; 3) non-melanoma cancer within 5 years; 4) persistent infection; 5) received systemic therapy within 2 years patients with autoimmune diseases;

6) Resected melanoma patients who have received systemic adjuvant therapy; 7) Patients who have previously received anti-PD-1, anti-PD-L1 or anti-PD-L2 immunotherapy. .

1.2 Clinical design

A total of 148 patients were planned to be enrolled and randomized 1:1 to the toripalimab group (test group) or interferon group (control group). Random stratification factors were disease stage (I vs II vs III), site of primary lesion (head and neck, non-head and neck), and PD-L1 expression status (positive, negative). Among them, the disease stage is defined according to the previous two studies (Lian B et al., Ann Oncol2017 28:868-873; Cui C et al., Ann Surg Oncol 2018 25:2184-2192): T1, mucosa or submucosa invasion; T2, Invasion of muscularis propria; T3, invasion of adventitia; T4, invasion of adjacent structures; N0, no regional nodes; N1, one or more regional lymph nodes; stage I, T1-2N0; stage II, T3–4/xN0; Stage III, T1-4/xN1.

Patients with head and neck mucosal melanoma (mainly including nasopharynx and oropharynx) were randomized into the toripalimab group or the HDI group. Patients in the experimental group will receive toripalimab group infusion (3mg/kg ivgtt q2w) for a maximum of 1 year (27 treatments). The patients in the control group will receive interferon α-2b. According to the patient's tolerance, some patients can choose appropriate low-dose escalation therapy (poor tolerance: 3MIU/m ² /d1, 6MIU/m ² /d2, 9MIU/ m ² /d3, well tolerated: 9MIU d1 or directly 15MIU/m ² /d). Follow-up received 15MIU/m ² /d, 1-5 days/week, for 4 weeks; then 9MIU/m ² /d, 3 times a week, for 48 weeks. Patients with head and neck mucosal melanoma should start receiving recombinant humanized anti-PD-1 monoclonal antibody injection or high-dose interferon therapy within 6-8 weeks, and start adjuvant radiation therapy at the same time. Reference dose: CTVTB 60-64Gy/ 30 times, 1-5 days/week, a total of 6 weeks.

Patients with non-head and neck mucosal melanoma were randomly divided into toripalimab group or HDI group. Patients in the experimental group will receive toripalimab group infusion (3mg/kg ivgtt q2w) for a maximum of 1 year (27 treatments). The patients in the control group will receive interferon α-2b. According to the patient's tolerance, some patients can choose appropriate low-dose escalation therapy (poor tolerance: 3MIU/m ² /d1, 6MIU/m ² /d2, 9MIU/ m ² /d3, well tolerated: 9MIU/m ² /d or directly 15MIU/m ² /d), followed by 15MIU/m ² /d on days 1-5/week for 4 weeks; then 9MIU/m 2 /d d, 3 times a week for 48 weeks.

Safety was evaluated during treatment. The treatment study can be terminated when there is recurrence and metastasis of the disease, unacceptable toxicity, withdrawal of informed consent or meeting other criteria for withdrawal from the study.

All enrolled subjects will be followed up for overall survival results (tumor re-examination assessment time: every 12 weeks in the first year, every 16 weeks in the second year, every six months in the 3rd to 5th years, and every year thereafter), and collect their Information on subsequent antineoplastic treatment and survival. If the subject terminates the study treatment without documented evidence of disease recurrence and metastasis, the tumor will continue to be assessed at the same frequency until the tumor relapses and metastasis or the subject begins to receive new anti-tumor therapy, whichever occurs first allow. The primary endpoint was RFS in the intention-to-treat population. Secondary endpoints included DMFS, 2-year RFS rate, OS and safety.

1.3 Number of patients

From July 2017 to May 2019, 187 patients were screened, and 145 patients were randomized 1:1 into the toripalimab group (n=73, test group) or interferon group (n=72 , control group). The demographic data of the enrolled subjects are shown in Table 1.

Table 1: Demographics and Baseline Clinical Characteristics of Study Subjects

PD-L1 positive: defined as CPS ≥ 1% of tumor cells stained by JS311 IHC

HDI, high-dose interferon-α2b; IQR, interquartile range; T, tumor stage; N, nodal stage; PD-L1, programmed death-ligand 1; CLND, complete lymph node dissection; LDH, lactate dehydrogenase; ULN, upper limit of normal;

1.4 Test drugs

Experimental drug: Recombinant humanized anti-PD1 monoclonal antibody injection (JS001, common name: Toripalimab injection, product name: Tuoyi) was provided by Shanghai Junshi Biomedical Technology Co., Ltd./Suzhou Zhonghe Biotechnology Co., Ltd. Medical Technology Co., Ltd. provides free; specification 240mg/6ml/bottle.

Control drug: Interferon α-2b injection was provided by Merck (Ganleneng), with a specification of 18MIU/1.2ml/bottle.

1.5 Disease assessment

Disease assessments were performed every 12 weeks for the first year, every 16 weeks for 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 nodal metastases detected by ultrasonographic findings of superficial lymph nodes should be verified histologically by lymphadenectomy or dissection, if necessary.

1.6 Efficacy and Safety Evaluation

Efficacy analyzes were performed on all randomized patients. The primary endpoint, RFS, refers to the time from the date of random enrollment to any documented tumor recurrence, metastasis or death.

Safety assessments were performed on all patients who received at least one dose of study treatment. Adverse events were assessed and recorded according to the National Cancer Institute Common Terminology Criteria, version 4.0.

1.7 PD-L1 expression analysis in tumor biopsy

Tumor PD-L1 IHC staining was performed in a 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 CellLung Cancer: A Phase 1 Trial. JAMA Netw Open 2020 3:e2013770). PD-L1 positivity was defined as the proportion of tumor cells and/or infiltrating immune cells CPS ≥ 1%.

1.8 Statistical Analysis

Validity:

For the primary efficacy endpoint, recurrence-free survival (RFS): refers to the time from the date of random enrollment to any recorded tumor recurrence, metastasis or death.

The survival time will be calculated using the non-parametric Kaplan-Meier method; if you want to study the impact of stratification factors such as disease stage on the survival curve, you can also use the stratified Kaplan-Meier method for analysis. The comparison of the survival curves of the two groups will use the two-sided Log-rank test.

Patients with no observed events (eg, relapse) were censored and their survival time was defined as the time between study initiation and last follow-up.

In addition, Cox proportional hazards regression was used for multivariate analysis to explore the research factors that may affect the recurrence-free survival curve.

For OS endpoints, Kaplan-Meier (KM) curves and median estimates will be provided as appropriate.

safety:

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

Summary statistics (number, percentage, median, standard deviation, etc.) for safety endpoints will be provided as appropriate. For any AE, any serious AE, any Grade 3-5 AE, any drug-related AE, any serious drug-related AE, any Grade 3-5 drug-related AE, dose suspension due to AE, drug withdrawal due to AE, any immunization The incidence of related AEs (irAEs), deaths, and specific AEs will provide summary statistics in numbers, percentages, and 95% confidence intervals.

Summary statistics will be provided for the mean and standard deviation of changes from baseline in laboratory tests and vital signs for each treatment cycle. Summary statistics will be provided for the number and percentage deterioration of laboratory values from baseline according to CTCAE grade.

2 Research results

From July 2017 to May 2019, 187 patients were screened, and 145 patients were randomized 1:1 into the toripalimab group (n=73, test group) or interferon group (n=72 , control group). Mean patient age was 58 years; most patients were female (62.8%); localized disease 80.7%, localized lymphoid disease 19.3%, local excision ± CLND 37.2%, wide excision ± CLND 62.8%; PD-L1 positive 51.0% (CPS≥1,22C3), PD-L1 negative 49.0%. Baseline characteristics did not differ between the two groups.

Among 57 (39.3%) patients with head and neck primary tumors, 24 (85.7%) of 28 patients in the HDI group and 26 (89.7%) of 29 patients in the toripalimab group received Adjuvant radiotherapy. A total of 52 patients completed the 1-year treatment period, including 30 (41.1%) patients in the toripalimab group and 22 (30.6%) patients in the HDI group. The current research process is shown in Figure 5.

As of March 31, 2021, the median follow-up time was 26.4 months in the HDI arm and 25.5 months in the toripalimab arm. As the first clinical study comparing immunotherapy with HDI in the treatment of mucosal melanoma, the results of this study show that toripalimab and HDI as adjuvant therapy have similar RFS in patients with completely resected mucosal melanoma, and toripalil The monoclonal antibody can not only significantly prolong the RFS of the PD-L1 positive subgroup, but at the same time, the safety and tolerability of toripalimab is significantly better than that of the HDI group, and the incidence of ≥ grade 3 treatment-related adverse events (TRAEs) Lower, reaching the pre-set superiority margin for the primary endpoint.

The results of the primary efficacy endpoint and various secondary efficacy endpoints are as follows:

2.1 Anti-tumor effect research

2.1.1 RFS, DMFS, OS values

As of March 31, 2021, among all 145 patients, a total of 97 RFS events occurred, including 46 RFS (63.9%) in the HDI group and 51 RFS (69.9%) in the toripalimab group. Median RFS values were similar in the HDI group and toripalimab group, 13.9 months (95% CI: 8.28-19.61) in the HDI group and 13.6 months (95% CI: 8.31) in the toripalimab group -19.02), the recurrence hazard ratio (HR) was 1.05 (95% CI: 0.69-1.61; stratified p=0.812) (Figure 1 and Table 2). In the HDI group and toripalimab group, the 1-year RFS rates were 52% (95% CI: 38.61–63.76) and 52.9% (95% CI: 40.33–63.90), and the 2-year RFS rates were 25.1% ( 95% CI: 14.75–36.83) and 30.5% (95% CI: 19.73–41.89). The recurrence hazard ratio HR of each subgroup, that is, the ratio of RFS events between the test group and the control group, is as described in Figure 2.

Table 2: Median RFS, DMFS and OS data of subjects (as of March 31, 2021)

PD-L1+ is defined as CPS ≥ 1% of tumor cells stained by JS311 IHC;

NR means not able to evaluate; NR: Not reached;

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

Table 3: Proportion of Patients Receiving Subsequent Immunotherapy

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

A total of 93 DMFS events occurred, including 44 (61.1%) in the HDI group and 49 (67.1%) in the toripalimab group. The median DMFS values of HDI group and toripalimab group were 14.6 months (95% CI: 8.34-21.26) and 16.3 months (95% CI: 10.94-21.09), respectively, and the hazard ratio of distant metastasis was 1.00 (95% CI: 0.65-1.54; stratified p=0.994). The 1-year DMFS rates of the HDI group and 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% ( 95CI: 16.20–39.18) and 32.0% (95CI: 20.86–43.67).

The median OS was not reached in the HDI group (95% CI: 28.29–NR), and the median OS in the toripalimab group was 35.1 months (95% CI: 27.93–NR) (Table 2). The relative hazard ratio (HR) for death was 1.11 (95% CI: 0.66–1.84). The HR of OS in each subgroup is shown in Figure 3.

2.1.2 Subgroup Efficacy

1) PD-L1 expression positive/negative subgroup

As of March 31, 2021, tumors positive for PD-1 expression were found in 36 (50.0%) patients in the HDI group and 38 (52.1%) patients in the toripalimab group. Among PD-L1 positive patients, the median 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, and the hazard ratio HR 1.00 (95% CI: 0.57–1.74; p=0.988), toripalimab could significantly prolong RFS in the PD-L1 positive subgroup (Table 2 and Figure 4A).

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

2) Other subgroups

For patients with disease stages I, II and III, the median RFS values in the HDI group were 8.3 months (95% CI: 5.52-21.29), 14.0 months (95% CI: 5.65-21.26) and 21.7 months, respectively. months (95%CI: 10.25-26.58); the median RFS values of the toripalimab group were 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). In addition, no significant differences in RFS were found by sex, age, tumor primary site, or other baseline characteristics.

2.2 Safety and Tolerability Studies

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

Table 4: Adverse events during treatment were reported in at least 15% of patients in both trials ^#

"^#" indicates a "common treatment-related adverse event" defined as occurring in ≥20% of patients in the toripalimab group.

Overall, in this first randomized phase II trial comparing toripalimab and HDI in patients with completely resected mucosal melanoma, median RFS was numerically superior to that of resection alone with both therapies. The results of the trial showed that toripalimab significantly prolonged RFS in the PD-L1 positive subgroup, highlighting the potential utility of these two interventions as adjuvant therapy for mucosal melanoma. Meanwhile, compared with HDI, toripalimab has superior safety and tolerability, which indicates that toripalimab is a better treatment option.

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Claims (10)

1. Use of an anti-PD-1 antibody or an antigen-binding fragment thereof in the preparation of a drug or kit for adjuvant treatment of a patient with completely resected mucosal melanoma or prevention of recurrence or distant metastasis of the patient's mucosal melanoma. 2. The use according to claim 1, wherein the mucosal melanoma is head and neck mucosal melanoma or non-head and neck mucosal melanoma. 3. The use according to claim 1 or 2, wherein the mucosal melanoma is a mucosal melanoma with positive expression of PD-L1 in immunohistochemical staining analysis of tumor tissue sections. 4. The use according to claim 1, wherein the mucosal melanoma is head and neck mucosal melanoma, and the use further comprises administering one or more therapies to individuals in need; preferably, The therapy is a chemotherapeutic agent, a biotherapeutic agent, an immunogenic agent, an immunostimulatory cytokine, an encoded immunostimulatory cytokine, or radiation therapy. 5. The use according to any one of claims 1-4, characterized in that the anti-PD-1 antibody or antigen-binding fragment thereof comprises amino acid sequences as shown in SEQ ID NO: 1, 2 and 3. a chain complementarity determining region, and a heavy chain complementarity determining region whose amino acid sequence is shown in SEQ ID NO: 4, 5 and 6; Preferably, the anti-PD-1 antibody or antigen-binding fragment thereof 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 whose amino acid sequence is shown in SEQ ID NO:9, and a heavy chain whose amino acid sequence is shown in SEQ ID NO:10. 6. The use according to any one of claims 1-4, wherein the anti-PD-1 antibody is selected from one or more of nivolumab, pembrolizumab, toripalimab, sintilimab, camrelizumab, tislelizumab, cemiplimab ; preferably toripalimab. 7. The use according to any one of claims 1-6, 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, 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 a fixed dose selected from about 120 mg to about 480 mg , for example about 120 mg, 240 mg, 360 mg or 480 mg fixed dose, preferably about 3 mg/kg body weight of an individual or about 240 mg fixed dose; Preferably, the anti-PD-1 antibody or 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 two weeks; Preferably, the anti-PD-1 antibody or antigen-binding fragment thereof is administered at a dose of 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, every two weeks or Apply once every three weeks. 8. The use according to any one of claims 1-7, wherein the anti-PD-1 antibody or its antigen-binding fragment is administered in a liquid dosage form such as an injection via a parenteral route such as intravenous infusion . 9. The use according to any one of claims 1-8, wherein the administration cycle of the anti-PD-1 antibody or its antigen-binding fragment is one week, two weeks, three weeks, one month, two weeks Months, three months, four months, five months, half a year, one year or more, optionally, the time of each administration cycle is the same or different, and the interval between each administration cycle is the same or different. 10. The use according to claim 4, characterized in that the dose of radiotherapy is CTV _TB 60-64Gy/30 times, administered on the 1st-5th day of each week, and administered for 6 weeks in total; preferably, in anti-PD -1 antibody or antigen-binding fragment thereof is administered within 6-8 weeks concurrently with radiation therapy.

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