TWI814752B - Uses of immunotherapy agents, nucleoside antimetabolites combined with platinum in the preparation of drugs for treating tumor - Google Patents

Abstract

The present invention provides to uses of immunotherapy agents, nucleoside antimetabolites combined with platinum in the preparation of drugs for treating tumor. Specifically, the present invention provides to uses of PD-1 antibody, gemcitabine combined with cisplatin in the preparation of drugs for treating nasopharyngeal carcinoma.

Description

Use of immunotherapeutic agents, nucleoside antimetabolites and platinum combinations in the preparation of drugs for the treatment of tumors

The present invention provides the use of a combination of an immunotherapeutic agent, a nucleoside antimetabolite and a platinum complex in the preparation of a drug for treating nasopharyngeal cancer.

Nasopharyngeal carcinoma (NPC) is a common malignant tumor in southern my country and Southeast Asia. It is highly malignant and metastasizes early. About 30% to 50% of nasopharyngeal cancer patients have cervical lymph node metastasis when they seek treatment. and distant transfer. With the gradual improvement of diagnosis and treatment methods of nasopharyngeal cancer, the local control rate of nasopharyngeal cancer patients has been significantly improved, but local recurrence and distant metastasis are still the main reasons for treatment failure of nasopharyngeal cancer. Radiotherapy or concurrent chemoradiotherapy is still the main treatment method for early-stage or locally advanced nasopharyngeal cancer. The 5-year survival rate is about 85%. However, these patients will eventually experience recurrence or metastasis, and 15% of nasopharyngeal cancer patients are diagnosed when they are first diagnosed. For distant metastasis, treatment for patients with recurrent or original metastatic nasopharyngeal carcinoma is very limited, and the median survival is mostly about 20 months.

Cancer immunotherapy is a hot topic in the field. It kills tumors by fully utilizing and mobilizing killer T cells in tumor patients. PD-1 (programmed death-1, programmed death receptor 1) antibodies can specifically recognize and bind to PD-1 on the surface of lymphocytes, block the PD-1/PD-L1 signaling pathway, and then activate T cell immunity against tumors. Killing effect, mobilize the body's immune system and eliminate tumor cells in the body. Epigenetic changes are closely associated with cancer development and drug resistance. As the research deepened, it was discovered that drug resistance existed after repeated administration of PD-1 antibodies to patients. Currently, many multinational pharmaceutical companies are developing monoclonal antibodies against PD-1. By blocking the binding between PD-L1/PD-1, it maximizes the patient's own response to the tumor immune system, thereby achieving the goal of treating tumors. cells for killing purposes. The PD-1 monoclonal antibodies disclosed by BMS, Merck, and WO2015085847 (publication date 2015.06.18) are currently cutting-edge PD-1 antibody drugs. The FDA has approved PD-1 antibodies for the treatment of malignant melanoma, non-small cell lung cancer, For single-agent use in multiple tumors such as head and neck squamous cell carcinoma, renal cancer, liver cancer, lymphoma, etc., the FDA has not yet approved PD-1 antibody monotherapy for the treatment of nasopharyngeal cancer, but Novartis' PDR-001 antibody is being used as a monotherapy for the nasopharyngeal cancer. Cancer clinical research.

Gemcitabine is a difluoronucleoside antimetabolite anticancer drug that damages cell replication. It is a water-soluble analog of deoxycytidine and an inhibitory enzyme substitute for ribonucleotide reductase. This enzyme is crucial for the generation of deoxyribonucleotides during DNA synthesis and repair. Cisplatin is a commonly used metal platinum complex, which has a broad anti-tumor spectrum and is effective against hypoxic cells. But it is toxic to the kidneys, nervous system and pancreas. Can combine with DNA to form cross bonds, thereby destroying the function of DNA It can no longer replicate; it also inhibits the synthesis of RNA and protein at high concentrations. It is a cycle-non-specific drug. Another feature of this product is that it also has an effect on hypoxic cells. After entering the human body, it can diffuse through charged cell membranes. It is relatively stable under the condition of high Cl- ion concentration. After entering the cell, due to the low intracellular Cl- concentration, the drug is hydrolyzed into a cationic hydrate, which has the effect of a bifunctional group similar to an alkylating agent and mainly interacts with the alkali on the DNA chain. base effect.

PD-1 monoclonal antibody combined with chemotherapy has achieved good results in non-small cell lung cancer and has been approved by the FDA. In addition, research on bladder cancer and breast cancer is also ongoing, but PD-1 monoclonal antibody combined with chemotherapy is rarely reported in nasopharyngeal cancer. We hope to further effectively treat nasopharyngeal cancer through PD-1 combined with chemotherapy.

The technical problem to be solved by the present invention is to provide a combined use of an immunotherapeutic agent, a nucleoside antimetabolite and a platinum complex in the preparation of a drug for the treatment of nasopharyngeal carcinoma. The combined treatment method shows good safety and efficacy. It has the advantages of high toxicity and low adverse reactions.

The technical solution of the present invention is as follows: The present invention provides the use of an immunotherapeutic agent, a nucleoside antimetabolite and a platinum complex in combination in the preparation of a drug for the treatment of nasopharyngeal cancer. The immunotherapeutic agent is selected from PD-1 antibodies. .

PD-1 antibodies are known. Preferably, the light chain variable region of the PD-1 antibody includes LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6 respectively. .

The heavy chain variable region of the PD-1 antibody includes SEQ ID NO: 1, HCDR1, HCDR2 and HCDR3 shown in SEQ ID NO:2 and SEQ ID NO:3.

Among them, the CDR sequences mentioned above are shown in the following table:

Preferably, the PD-1 antibody is a humanized antibody.

The preferred humanized antibody light chain variable region sequence is the sequence shown in SEQ ID NO: 10 or a variant thereof. The variant preferably has 0 to 10 amino acid changes in the light chain variable region, More preferably, it is the amino acid change of A43S; the humanized antibody heavy chain variable region sequence is the sequence shown in SEQ ID NO: 9 or a variant thereof, and the variant preferably has 0 in the heavy chain variable region. The amino acid change is to 10, more preferably the amino acid change of G44R.

SEQID NO：9 The variable region sequences of the heavy and light chains of the aforementioned humanized antibodies are as follows: Heavy chain variable region

SEQ ID NO: 9

SEQID NO：10 light chain variable region

SEQ ID NO: 10

The preferred humanized antibody light chain sequence is the sequence shown in SEQ ID NO: 8 or a variant thereof; the variant preferably has 0 to 10 amino acid changes in the light chain variable region; more preferably Amino acid changes in A43S.

The humanized antibody heavy chain sequence is the sequence shown in SEQ ID NO: 7 or a variant thereof; the variant preferably has 0 to 10 amino acid changes in the heavy chain variable region; more preferably it is G44R. Amino acid changes.

A particularly preferred light chain sequence of the humanized antibody is the sequence shown in SEQ ID NO: 8, and the heavy chain sequence is the sequence shown in SEQ ID NO: 7.

SEQID NO：7 The sequences of the heavy and light chains of the aforementioned humanized antibodies are as follows: Heavy chain

SEQ ID NO: 7

SEQID NO：8 light chain

SEQ ID NO: 8

In a preferred embodiment of the present invention, the nucleoside anti-metabolite is selected from gemcitabine, mercaptopurine, nelarabine, fludarabine, cladribine, clofarabine ), azacitidine, amcitabine, troxacitabine, capecitabine, decitabine, preferably gemcitabine.

In some embodiments, the platinum-based complex is selected from cisplatin, carboplatin, nedaplatin, oxaliplatin, or loplatin, preferably cisplatin.

In a preferred embodiment of the present invention, the tumor is selected from malignant tumors and benign tumors; the malignant tumor is selected from malignant epithelial tumors, sarcoma, myeloma, leukemia, lymphoma, melanoma, head and neck tumors, brain tumors, Mixed tumors, childhood malignant tumors; the malignant epithelial tumors are selected from lung cancer, breast cancer, liver cancer, pancreatic cancer, colorectal cancer, gastric cancer, gastroesophageal adenocarcinoma, gastroesophageal adenocarcinoma, Tube cancer, small bowel cancer, cardia cancer, endometrial cancer, ovarian cancer, fallopian tube cancer, vulva cancer, testicular cancer, prostate cancer, penis cancer, kidney cancer, bladder cancer, anal cancer, gallbladder cancer, cholangiocarcinoma, teratoma , heart tumor; the head and neck tumor is selected from nasopharyngeal cancer, laryngeal cancer, thyroid cancer, tongue cancer, oral cancer; the sarcoma is selected from Askin tumor, chondrosarcoma, Ewing's sarcoma, malignant hemangioendothelioma, malignant schwannoma, Osteosarcoma, soft tissue sarcoma; the myeloma is selected from the group consisting of solitary myeloma, multiple myeloma, diffuse myeloma, leukemic myeloma, extramedullary myeloma; the leukemia is selected from the group consisting of acute lymphoblastic leukemia, chronic lymphocytic leukemia , acute myeloid leukemia, chronic myelogenous leukemia, hairy cell leukemia, T-cell lymphocytic leukemia, large granular lymphocytic leukemia, adult T-cell leukemia; the lymphoma is selected from the group consisting of non-Hodgkin lymphoma, Hodgkin lymphoma tumor; the brain tumor is selected from neuroepithelial tissue tumors, cranial nerve and spinal cord nerve tumors, and meningeal tissue tumors; the childhood malignant tumor is selected from Wilms tumor, neuroblastoma, retinoblastoma, and childhood germ cell tumors. .

In another preferred embodiment of the present invention, the nasopharyngeal carcinoma is selected from tumors that have not received any chemotherapy before, are recurrent, metastatic, are not suitable for local treatment, have recurred or metastasized after radiotherapy, and preferably have not received any chemotherapy before. , Not suitable for local treatment of nasopharyngeal carcinoma, recurrence or metastasis of nasopharyngeal carcinoma after radiotherapy.

In a preferred embodiment of the present invention, the tumor is resistant or drug-resistant to immunotherapeutic agents or immunotherapy. Preferably, the immunotherapeutic agent is based on PD-1 and/or PD-L1 or CTLA-4. (Cytotoxic T lymphocyte-associated protein 4) is the target; the immunotherapy is selected from immune checkpoint blockade (ICB) therapy, chimeric antigen receptor T cell immunotherapy (CAR-T therapy), autologous cell immune therapy Immunotherapy (CIK therapy).

In a preferred embodiment of the present invention, preferably, the immunotherapeutic agent is selected from the group consisting of PD-1 antibodies, PD-L1 antibodies, and CTLA-4 antibodies. The PD-1 antibodies include but are not limited to pidilizumab ( Pidilizumab), MEDI-0680, AMP-224, PF-06801591, TSR-042, JS-001, GLS-010, PDR-001, Genolimzumab, Camrelizumab, BGB-A317, IBI-308, REGN-2810, Pembrolizumab, Nivolumab; the PD-L1 antibodies include but are not limited to MSB-0011359-C, CA-170, LY- 3300054, BMS-936559, Durvalumab, Avelumab, Atezolizumab; the CTLA-4 antibodies include but are not limited to ipilimumab , AK-104, JHL-1155, ATOR-1015, AGEN-1884, PRS-010, tremelimumab, IBI-310, MK-1308, BMS-986218, SN-CA21, FPT-155, KN -044, CG-0161, ONC-392, AGEN-2041, PBI-5D3H5.

In a preferred embodiment of the present invention, the invention provides the use of PD-1 antibodies in combination with nucleoside antimetabolites and platinum in the preparation of drugs that enhance the activity of T-cells, preferably peripheral T-cells.

In a preferred embodiment of the present invention, the dose of the immunotherapeutic agent is selected from 1-10 mg/kg, preferably 1 mg/kg, 2 mg/kg, 3 mg/kg, 4 mg/kg, 5 mg/kg, 6 mg/kg, 7 mg /kg, 8mg/kg, 9mg/kg, 10mg/kg, preferably 3mg/kg, 4mg/kg, 5mg/kg.

In a preferred embodiment of the invention, the immunotherapeutic agent dose is selected from the group consisting of 50-600mg, preferably 50mg, 60mg, 70mg, 75mg, 100mg, 125mg, 150mg, 175mg, 200mg, 225mg, 250mg, 375mg, 400mg, 425mg, 450mg, 475mg, 500mg, 600mg, more preferably 100mg, 200mg, 400mg.

In a preferred embodiment of the present invention, the nucleoside anti-metabolite dosage is selected from 5-1000 mg/m ² , preferably from 5 mg/m ² , 6 mg/m ² , 7 mg/m ² , 8 mg/m ² , 9mg/m ² , 10mg/m ² , 12mg/m ² , 15mg/m ² , 20mg/m 2 , 25mg/m ² , 30mg/m ² , 35mg/m ² , 40mg/ ^{m 2 ,} 50mg/m ² , 60mg/m ² , 75mg/m ² , 80mg/m ² , 85mg/m 2 , 90mg/m ² , 100mg/ ^{m 2 ,} 110mg/m ² , 150mg/m ² , 180mg/m ² , 200mg/m ² , 250mg/m ² , 300mg/m ² , 350mg/m ² , 400mg/m ^{2 , 450mg/m 2 ,} 500mg/m ² , 550mg/m ² , 600mg/m ² , 650mg/m ² , 700mg/m ² , 800mg/m ² , 900mg/m ² , 1000mg/m ² ; better 7mg/m ² , 10mg/m ² , 12mg/m ² , 15mg/m ² , 20mg/m ² , 25mg/m ² , 30mg/m ^2. 40mg/m ² , 50mg/m ² , 75mg/m ² , 100mg/m ² , 150mg/m ² , 180mg/m ² , 200mg/m ² , 300mg/m ² , 400mg/m ² , 500mg/m ² , 600mg/m ² , 700mg/m ² , 800mg/m ² , 900mg/m ² or 1000mg/m ² .

In a preferred embodiment of the present invention, the nucleoside anti-metabolite dosage is selected from 5-2000 mg, preferably from 10 mg, 15 mg, 20 mg, 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 225 mg , 250mg, 275mg, 300mg, 400mg, 500mg, 600mg, 800mg, 100mg, 1200mg, 1500mg, 1600mg, 1700mg, 1800mg, 2000mg, more preferably 10mg, 20mg, 100mg, 200mg, 300mg, 500mg, 1000mg or 1500mg.

In a preferred embodiment of the present invention, the dosage of the platinum complex is selected from 5-200 mg/m ² , preferably from 5 mg/m ² , 6 mg/m ² , 7 mg/m ² , 8 mg/m ² , 9 mg /m ² , 10mg/m ² , 12mg/m ² , 15mg/m ² , 20mg/m 2 , 25mg/m ² , 30mg/m ² , 35mg/m ² , 40mg/ ^{m 2 ,} 50mg/m ² , 60mg /m ² , 75mg/m ² , 80mg/m ² , 85mg/m 2 , 90mg/m ² , 95mg/m ² , 100mg/ ^{m 2 ,} 120mg/m ² , 125mg/m ² , 130mg/m ² , 150mg /m ² , 160mg/m ² , 180mg/m ² , 180mg/m ² , 190mg/m ² , 195mg/m ² , 200mg/m ² ; better 7mg/m ² , 10mg/m ² , 12mg/m ² , 15mg/m ² , 20mg/m ² , 25mg/m ² , 30mg/m ² , 40mg/m ² , 50mg/m ² , 75mg/m ² , 80mg/m ² .

In a preferred embodiment of the present invention, the dosage of the platinum complex is selected from 5-400 mg, preferably from 10 mg, 15 mg, 20 mg, 25 mg, 40 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg , 85mg, 90mg, 95mg, 100mg, 125mg, 130mg, 135mg, 150mg, 155mg, 160mg, 165mg, 170mg, 180mg, 185mg, 190mg, 200mg, 250mg, 300mg, 400mg, preferably 10mg, 20mg, 50mg, 60mg, 70mg , 80mg, 100mg.

In a preferred embodiment of the present invention, the combination further includes a fourth component selected from the group consisting of alkylating agents, platinum complexes, nucleoside anticancer agents, metabolic antagonists, plant alkaloids, One or more of hormonal anticancer agents, proteasome inhibitors, aromatase inhibitors, immunomodulators, EGFR inhibitors, ALK inhibitors, PARP inhibitors, VEGF antibodies, VEGFR inhibitors, and mTOR inhibitors. Preferably, the chemotherapeutic drug is selected from the group consisting of alkylating agents, platinum complexing agents, metabolic antagonists, plant alkaloids (such as vinblastine, H. ester bases), hormone anti-cancer agents, proteasome inhibitors, aromatase inhibitors, immunomodulators, one or more; in another preferred embodiment, the chemotherapeutic drugs include but are not limited to cyclophosphamide, Ifosfamide, melphalan, busulfan, nimustine, ramustine, dacarbazine, temozolomide, nitrogen mustard, dibromomannitol, cisplatin, carboplatin, oxaliplatin, Nedaplatin, methotrexate, 5-fluorouracil, tegafur, decitabine, capecitabine, fulvestrant, pemetrexed, anthracyclines, mitomycin, bleomycin antibiotics, actinomycins, vinblastines, camptothecins, paclitaxels, vincristine, vinblastine, vindesine, etoposide, docetaxel, paclitaxel, albumin-bound paclitaxel, paclitaxel liposomes , irinotecan, vinorelbine, mitoxantrone, vinflunine, topotecan, leuprorelin, goserelin, dutasteride, fulvestrant, dexamethasone, tamoxifen , bortezomib, lenalidomide, etc., exemestane, letrozole, anastrozole.

Preferably, the targeted drug is selected from one or more therapeutic agents selected from the group consisting of EGFR inhibitors, ALK inhibitors, PARP inhibitors, VEGF antibodies, VEGFR inhibitors, and mTOR inhibitors. These targeted drugs are well known in the art. For example, EGFR inhibitors are selected from gefitinib, erlotinib, icotinib, afatinib, cetuximab, and trastuzumab. One or more; ALK inhibitor is selected from one or more of crizotinib, ceritinib, axitinib, and brigatinib; VEGF antibody is selected from bevacizumab, ibuprofen One or more of Brolucizumab, Vanucizumab, Navicixizumab, Ranibizumab, Conbercept; VEGFR inhibition The agent is selected from sunitinib, apatinib, and famitinib. species or several species.

In some embodiments, the fourth component is selected from decitabine, carboplatin, paclitaxel albumin, paclitaxel liposome, paclitaxel, docetaxel, cyclophosphamide, doxorubicin, tetrahydrouridine (Tetrahydrouridine).

In the above-mentioned preferred embodiment of the present invention, the fourth component can be determined according to the patient's body surface area, weight, or KPS functional status scoring standard or ECOG performance status scoring standard (Zubrod-ECOG-WHO) and various tumor diagnosis and treatment guidelines. Choose the recommended dosage and dosage regimen for different types of tumor chemotherapy regimens. For example, the dosage of paclitaxel and albumin is 50-500 mg/m ² , preferably 125 mg/m ² ; the dosage of decitabine is 5 to 500 mg/m ² , preferably 5 to 100 mg/m ² .

In the present invention, the treatment cycle can be 1 day, 3 days, 1 week, 2 weeks, or 3 weeks, preferably 3 weeks.

In the present invention, the treatment cycle includes but is not limited to chemotherapy cycle or radiotherapy cycle or other related targeted drug treatment cycle or immunotherapy cycle.

In the present invention, immunotherapeutic agents, nucleoside antimetabolites and platinum complexes can be used in combination to treat tumors in the same or different treatment cycles. In the process of treating tumors, immunotherapeutic agents, nucleoside antimetabolites and platinum complexes At the same time, before or after the combined administration of antimetabolites and platinum complexes, the tumors can also be treated with better chemotherapy regimens or radiotherapy regimens or targeted small molecule drug regimens or immunotherapy regimens based on different tumors. Treatment options include but are not limited to cellular immunotherapy (such as CAR-T therapy, tumor vaccines, CIK therapy, etc.); in addition, the combined administration of epiimmunotherapy agents, nucleoside antimetabolites, and platinum complexes may not be combined Other treatment options are performed individually.

In the present invention, immunotherapeutic agents, nucleoside antimetabolites and platinum complexes can be used in combination to carry out treatment plans for tumors of different pathological classifications and progression stages stipulated in various tumor diagnosis and treatment standards or guidelines. , the tumor diagnosis and treatment standards or guidelines include but are not limited to NCCN (National Comprehensive Cancer Network clinical practice guidelines for various malignant tumors) or the malignant tumor diagnosis and treatment standards promulgated by the Ministry of Health of China.

In the present invention, within one treatment cycle, the immunotherapeutic agent, the nucleoside antimetabolite and the platinum complex are administered simultaneously or before the nucleoside antimetabolite or after the nucleoside antimetabolite. , preferably, gemcitabine can be administered on the 1st, 2nd, 3rd, 4th, 5th, 6th and 7th days after the end of PD-1 antibody administration, preferably on the 1st, 2nd, 3rd, 4th, 5th, 6th and 7th days. Administer on day 1 and day 8. Cisplatin can be administered on day 1, day 2, day 3, day 4, day 5, day 6, and day 7 after the end of PD-1 antibody administration. The drug is preferably administered on the first day; every 3 weeks is a treatment cycle, and the combined drug can be used for up to 6 cycles. After that, the PD-1 antibody can be used alone to maintain treatment until the end.

In the present invention, the invention further provides uses in medicines, wherein the frequency of administration of the immunotherapeutic agent is once a day, twice a day, three times a day, once a week, once every two weeks, or once every three weeks. The administration frequency of glycoside antimetabolites is once a day, twice a day, three times a day, once a week, once every two weeks, once every three weeks, once a month, and once every two months. The administration frequency of platinum complexes is The frequency is once a day, twice a day, three times a day, once a week, once every two weeks, once every three weeks, and once a month.

In a preferred embodiment of the present invention, during administration, 3 weeks constitutes a cycle, wherein the dosage of the PD-1 antibody is 60 to 600 mg, intravenously infused, and administered on the first day of each cycle; nucleoside antibodies The dosage of metabolites is 200mg to 2000mg/m ² , intravenous infusion, administered on the 1st and 8th days of each cycle; the dosage of platinum complexes is 5 to 200mg/m ² , intravenous infusion, administered on the 1st day of each cycle Dosing; use for a maximum of 2 to 12 cycles, and then use PD-1 antibody alone to maintain treatment until disease progression, intolerable toxicity, subject requirements, or the investigator determines that the subject is not suitable to continue treatment.

In a preferred embodiment of the present invention, during administration, 3 weeks constitutes a cycle, wherein the dosage of the PD-1 antibody is 80 to 300 mg, intravenously infused, and administered on the first day of each cycle; nucleoside antibodies The dosage of metabolites is 500 to 1500 mg/m ² , intravenous infusion, administered on the 1st and 8th days of each cycle; the dosage of platinum complexes is 10 to 100 mg/m ² , intravenous infusion, administered on the 1st day of each cycle Dosing; use for a maximum of 3 to 10 cycles, and then use PD-1 antibody alone to maintain treatment until disease progression, intolerable toxicity, subject requirements, or the investigator determines that the subject is not suitable to continue treatment.

In a preferred embodiment of the present invention, during administration, 3 weeks constitutes a cycle, wherein the dosage of the PD-1 antibody is 200 mg, intravenously infused, and administered on the first day of each cycle; nucleoside antimetabolites The dosage of platinum complex is 1000mg/m ² , intravenous infusion, administered on the 1st and 8th day of each cycle; the dosage of platinum complex is 80mg/m ² , intravenous infusion, administered on the 1st day of each cycle; maximum use is 6 cycles, and then use PD-1 antibody alone for maintenance treatment until disease progression, intolerable toxicity, subject's request, or the investigator's judgment that the subject is not suitable to continue treatment.

In a preferred embodiment of the present invention, the immunotherapeutic agent is administered by injection, such as subcutaneous or intravenous injection. The immunotherapeutic agent needs to be administered before injection. Formulated in an injectable form. Particularly preferred injectable forms of immunotherapeutic agents are injection solutions or lyophilized powder injections, which contain the immunotherapeutic agent, a buffer, a stabilizer, and optionally a surfactant. The buffering agent may be selected from one or more of acetate, citrate, succinate and phosphate. The stabilizer can be selected from sugars or amino acids, preferably disaccharides, such as sucrose, lactose, trehalose, and maltose. The surfactant is selected from polyoxyethylene hydrogenated castor oil, glycerin fatty acid ester, and polyoxyethylene sorbitan fatty acid ester. Preferably, the polyoxyethylene sorbitan fatty acid ester is polysorbate 20, 40, 60 or 80. , the best is polysorbate 20. The optimal injectable form of the immunotherapeutic agent contains PD-1 antibody, acetate buffer, trehalose, and polysorbate 20.

The combined administration route of the present invention is selected from oral administration, parenteral administration, and transdermal administration. The parenteral administration includes but is not limited to intravenous injection, subcutaneous injection, and intramuscular injection.

The present invention provides a combination of the above immunotherapeutic agent, nucleoside antimetabolite and platinum complex as a therapeutic preparation for treating tumors and/or enhancing T-cell activity.

In the present invention, a method of treating tumors and/or enhancing T-cell activity is provided, which includes administering the above-mentioned immunotherapeutic agent, nucleoside antimetabolite and platinum complex to a patient.

The present invention also provides a drug set or a drug packaging box, which contains the aforementioned immunotherapeutic agent, nucleoside anti-metabolite and platinum complex.

(detailed description of the invention)

In order to make the present invention easier to understand, certain technical and scientific terms are specifically defined below. Unless otherwise clearly defined elsewhere in this document, all other technical and scientific terms used herein have the meaning commonly understood by one of ordinary skill in the art to which this invention belongs.

The term "humanized antibody", also known as CDR-grafted antibody, refers to transplanting the mouse CDR sequence into the human antibody variable region framework, that is, different types of human germline Antibodies produced from antibody framework sequences. It can overcome the strong antibody variable antibody response induced by chimeric antibodies carrying a large amount of mouse protein components. Such framework sequences can be obtained from public DNA databases or published references that include germline antibody gene sequences. For example, germline DNA sequences of human heavy and light chain variable region genes are available in the "VBase" human germline sequence database (available on the Internet at www.mrccpe.com.ac.uk/vbase), and in Kabat, E.A. et al. Found in Man, 1991 Sequences of Proteins of Immunological Interest, 5th ed. In a preferred embodiment of the present invention, the CDR sequence of the PD-1 humanized antibody is selected from SEQ ID NO: 1, 2, 3, 4, 5, 6.

The term "antigen-binding fragment" refers to Fab fragments, Fab' fragments, F(ab')2 fragments with antigen-binding activity, and Fv fragments that bind to human PD-1; sFv fragments that include the antibodies of the present invention are selected from One or more CDR regions of SEQ ID NO: 1 to SEQ ID NO: 6. Fv fragments contain the antibody heavy chain variable region and the light chain variable region, but no constant region, and have all The smallest fragment of an antibody that has an antigen-binding site. Typically, Fv antibodies also contain a polypeptide linker between the VH and VL domains and are capable of forming the structure required for antigen binding. Different linkers can also be used to connect the variable regions of two antibodies into a polypeptide chain, called a single chain antibody or single chain Fv (sFv). The term "binding to PD-1" in the present invention refers to the ability to interact with human PD-1. The term "antigen binding site" of the present invention refers to a discontinuous three-dimensional site on the antigen recognized by the antibody or antigen-binding fragment of the present invention.

The term "immunotherapy" means that immunotherapy uses the immune system to treat diseases. In the present invention, it mainly refers to stimulating and enhancing the body's anti-tumor immune response by improving the immunogenicity of tumor cells and their sensitivity to effector cell killing, and Immune cells and effector molecules are infused into the host body to coordinate with the body's immune system to kill tumors and inhibit tumor growth.

The following examples are used to further describe the present invention, but these examples do not limit the scope of the present invention.

Example 1: PD-1 antibody combined with gemcitabine and cisplatin in the treatment of recurrent or metastatic nasopharyngeal carcinoma

1. Test antibodies and compounds

The sequences of the heavy and light chains of the PD-1 antibody are as follows: SEQ ID NO: 7 and SEQ ID NO: 8 in the present invention. 200 mg/tube, formulated into 40 mg/ml for later use.

Gemcitabine is commercially available as freeze-dried powder for injection, with a specification of 1.0g/bottle. Please refer to the marketing instructions for the preparation method.

Cisplatin injection is commercially available in specifications of 20 ml: 20 mg. Please refer to the marketing instructions for the preparation method.

2. Enrolled subjects

(1) Patients with pathologically confirmed nasopharyngeal cancer (primary metastatic nasopharyngeal cancer or recurrent nasopharyngeal cancer not suitable for local treatment), patients who have not received systemic chemotherapy for recurrent or metastatic nasopharyngeal cancer in the past, and who are taking 6 drugs for the first time Except for neoadjuvant chemotherapy, concurrent chemotherapy or adjuvant chemotherapy received more than 6 months ago; have measurable lesions; ECOG score 0-1 points; (2) age 18 to 70 years old;

3. Administration method

PD-1 antibody (200 mg/time, on day 1 of each cycle, given at least 30 minutes before chemotherapy drugs, intravenous infusion >30 minutes), gemcitabine (1000 mg/m ² , on days 1 and 8 of each cycle, intravenous infusion for 30 minutes ) and cisplatin (80 mg/m ² , intravenous infusion for 4 hours on the first day of each cycle). Every 3 weeks is a treatment period. If the dosing date exceeds 3 days in each treatment cycle, it will be considered as delayed dosing. The subsequent dosing time will be calculated based on the actual date of the previous dosing. The combination therapy is used for up to 6 cycles, and then the PD-1 antibody is used alone for maintenance therapy until disease progression, intolerable toxicity, the subject's request, or the investigator's judgment that the subject is not suitable for continued treatment. As of October 27, 2017, 23 cases were enrolled and 22 were used for evaluation.

Experimental conclusion: As shown in Table 1, the three-component combination of PD-1 antibody, gemcitabine, and cisplatin in the treatment of recurrent or metastatic nasopharyngeal carcinoma has a response rate of 90.9% and a control rate of 100%, showing good safety and effectiveness. sex.

<110> Jiangsu Hengrui Pharmaceutical Co., Ltd. Suzhou Shengdia Biopharmaceutical Co., Ltd.

<120> Immunotherapeutic agents, nucleoside antimetabolites and platinum combinations in the preparation of drugs for the treatment of tumors the use of

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<400> 10

Claims (38)

A use of a PD-1 antibody, a nucleoside antimetabolite and a platinum complex, which are combined in the preparation of a drug for the treatment of nasopharyngeal cancer; wherein the light chain variable region of the PD-1 antibody includes SEQ ID LCDR1, LCDR2 and LCDR3 shown in NO:4, SEQ ID NO:5 and SEQ ID NO:6; the heavy chain variable region of the PD-1 antibody includes SEQ ID NO:1, SEQ ID NO:2 and HCDR1, HCDR2 and HCDR3 shown in SEQ ID NO: 3; the nucleoside anti-metabolite is selected from gemcitabine, mercaptopurine, nelarabine, fludarabine, cladribine, Clofarabine, azacitidine, amcitabine, trosatabine, capecitabine or decitabine, the platinum complex is selected from cisplatin, carboplatin, nedaplatin, oxalate thaliplatin or loplatin. The use described in item 1 of the patent application, wherein the PD-1 antibody is a humanized antibody. The use described in item 2 of the patent application, wherein the light chain variable region sequence of the humanized antibody is the sequence shown in SEQ ID NO: 10 or a variant thereof; the heavy chain variable region sequence is as follows The sequence shown in SEQ ID NO: 9 or a variant thereof. The use described in item 3 of the patent application, wherein the variant of the light chain variable region sequence of the humanized antibody has 0 to 10 amino acid changes in the light chain variable region. The use described in item 4 of the patent application, wherein the variant of the light chain variable region sequence of the humanized antibody is an amino acid change of A43S. The use described in item 3 of the patent application, wherein the variant of the heavy chain variable region sequence of the humanized antibody has 0 to 10 amino acid changes in the heavy chain variable region. The use described in item 6 of the patent application, wherein the variant of the heavy chain variable region sequence of the humanized antibody is an amino acid change of G44R. The use as described in item 3 of the patent application, wherein the humanized antibody light chain sequence is the sequence shown in SEQ ID NO: 8 or a variant thereof; the heavy chain sequence is as shown in SEQ ID NO: 7 sequence or a variant thereof. The use as described in item 8 of the patent application, wherein the variant of the light chain variable region sequence of the humanized antibody has 0 to 10 amino acid changes in the light chain variable region. The use as described in item 9 of the patent application, wherein the variant of the light chain variable region sequence of the humanized antibody is an amino acid change of A43S. The use as described in item 8 of the patent application, wherein the variant of the heavy chain variable region sequence of the humanized antibody has 0 to 10 amino acid changes in the heavy chain variable region. The use as described in item 11 of the patent application, wherein the variant of the heavy chain variable region sequence of the humanized antibody is an amino acid change of G44R. The use described in item 8 of the patent application, wherein the humanized antibody light chain sequence is the sequence shown in SEQ ID NO: 8, and the heavy chain sequence is the sequence shown in SEQ ID NO: 7. The use described in item 1 of the patent application, wherein the nucleoside anti-metabolite is gemcitabine. The use described in item 1 of the patent application, wherein the platinum complex The substance is cisplatin. The use as described in item 1 of the patent application, wherein the nasopharyngeal cancer is selected from tumors that have not received any chemotherapy before, are recurrent, metastatic, unsuitable for local treatment, relapsed or metastasized after radiotherapy. For the use described in Item 16 of the patent application, the nasopharyngeal cancer is selected from nasopharyngeal cancer that has not received any chemotherapy before, is not suitable for local treatment, and has recurred or metastasized after radiotherapy. The use described in item 16 of the patent application, wherein the dose of the PD-1 antibody is selected from 1 to 10 mg/kg. The use as described in item 18 of the patent application, wherein the dose of the PD-1 antibody is selected from 1 mg/kg, 3 mg/kg, 5 mg/kg, 8 mg/kg or 10 mg/kg. The use described in item 19 of the patent application, wherein the dose of the PD-1 antibody is selected from 1 mg/kg, 3 mg/kg or 10 mg/kg. The use described in item 18 of the patent application, wherein the dose of the PD-1 antibody is selected from 50 to 600 mg. The use as described in item 21 of the patent application, wherein the dose of the PD-1 antibody is selected from 50 mg, 60 mg, 70 mg, 100 mg, 150 mg, 200 mg, 300 mg, 400 mg, 500 mg or 600 mg. The use described in item 22 of the patent application, wherein the dose of the PD-1 antibody is selected from 60 mg, 200 mg or 400 mg. The use described in Item 16 of the patent application, wherein the dose of the nucleoside antimetabolite is selected from 5 to 2000 mg/m ² . The use as described in item 24 of the patent application, wherein the dose of the PD-1 antibody is selected from 5mg/m ² , 10mg/m ² , 20mg/m ² , 40mg/m ² , 80mg/m ² , 100mg/m ² , 300mg/m ² , 500mg/m ² , 800mg/m ² , 1000mg/m ² or 2000mg/m ² . The use as described in item 25 of the patent application, wherein the dose of the PD-1 antibody is selected from 100 mg/m ² , 500 mg/m ² , 800 mg/m ² or 1000 mg/m ² . The use described in item 24 of the patent application, wherein the dose of the nucleoside antimetabolite is selected from 5 to 2000 mg. The use as described in item 27 of the patent application, wherein the dose of the nucleoside antimetabolite is selected from 10 mg, 100 mg, 200 mg, 500 mg, 800 mg, 1000 mg, 1500 mg or 2000 mg. The use as described in item 28 of the patent application, wherein the dose of the PD-1 antibody is selected from 100 mg, 500 mg, 1000 mg or 1500 mg. The use described in item 16 of the patent application, wherein the dose of the platinum complex is selected from 5 to 200 mg/m ² . The use as described in item 30 of the patent application, wherein the dosage of the platinum complex is selected from 5mg/m ² , 10mg/m ² , 20mg/m ² , 50mg/m ² , 80mg/m ² , 100mg/ m ² , 150mg/m ² or 200mg/m ² . The use described in item 31 of the patent application, wherein the dosage of the platinum complex is selected from 10 mg/m ² , 50 mg/m ² or 80 mg/m ² . The use described in item 30 of the patent application, wherein the dose of the platinum complex is selected from 5 to 400 mg. The use described in item 33 of the patent application, wherein the dosage of the platinum complex is selected from 10 mg, 20 mg, 40 mg, 100 mg, 160 mg, 200 mg, 300 mg or 400 mg. The use described in item 34 of the patent application, wherein the dosage of the platinum complex is selected from 20 mg, 100 mg or 160 mg. A drug package for treating nasopharyngeal cancer, characterized in that it contains a PD-1 antibody, nucleoside antimetabolites and platinum complexes; the light chain variable region of the PD-1 antibody contains respectively as follows: LCDR1, LCDR2 and LCDR3 shown in SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6; the heavy chain variable region of the PD-1 antibody includes SEQ ID NO: 1 and SEQ ID NO: respectively. 2 and HCDR1, HCDR2 and HCDR3 shown in SEQ ID NO: 3; the platinum complex is selected from cisplatin, carboplatin, nedaplatin, oxaliplatin or loplatin, and the nucleoside antimetabolite is selected from From gemcitabine, mercaptopurine, nelarabine, fludarabine, cladribine, clofarabine, azacitidine, amcitabine, troxacitabine, carbo Petabine or decitabine. The use described in item 36 of the patent application, wherein the platinum complex is selected from cisplatin. The use described in item 36 of the patent application, wherein the nucleoside anti-metabolite is selected from gemcitabine.