TW201922282A - Combination use of PD-1 antibody and epigenetic modulating agent in the preparation of a medicament for the treatment of tumor - Google Patents

Abstract

The present invention relates to combination use of PD-1 antibody and epigenetic modulating agent in the preparation of a medicament for the treatment of tumor. In particular, the present invention relates to combination use of PD-1 antibody and epigenetic modulating agent in the preparation of a medicament for the treatment of tumor or enhancing T-cell activity.

Description

Application of PD-1 antibody and epigenetic modulator in preparing medicine for treating tumor

Use of a combination of an anti-PD-1 antibody and an epigenetic modulator in the manufacture of a medicament for treating tumors and / or enhancing T-cell activity.

PD-1 (programmed death receptor 1) antibody can specifically recognize and bind to PD-1 on the surface of lymphocytes, block the PD-1 / PD-L1 signaling pathway, and then activate the immune killing effect of T cells on tumors and regulate the body The immune system removes tumor cells in the body. Epigenetic changes are closely related to cancer development and drug resistance. With the deepening of research, it was found that drug resistance exists after repeated administration of PD-1 antibody to patients. DNA methylation inhibitor decitabine (Dacogen ^® ) has been approved for hematological treatment of malignant tumors and its clinical effect on solid tumors. Various animal models and cell line studies show that decitabine induces control Apoptosis, cell cycle arrest, and gene expression of tumor surface antigens; DNA demethylation effect induces major histocompatibility complex (MHC) and costimulatory molecules, resulting in increased decitabine resistance to tumors Response rate of immunotherapy, inhibit tumor growth. Studies have found that methylation modification and the silencing of specific genes may play a key role in the maturation and typing of T-cells. 5-azacitidine has been reported to increase PD-1 / PD-L1 (programmed death Body 1) is the target of the sensitivity of immune checkpoint therapy, so epigenetic therapy combined with immunotherapy can improve the effectiveness of clinical treatment of tumors (Cancer letters 354.1 (2014): 12-20). Studies by Riccadonna et al. Have shown that demethylating agents have improved immunotherapy's recognition and subsequent destruction of various glioma cells, and its cell and animal tests have shown that decitabine combined with multiple immunotherapy has good tumor suppression in a model of malignant glioma Effect ([J] .PloS one, 2016,11 (8): e0162105); study by Tamas A et al. Found decitabine combined with immune checkpoint inhibitor (Anti-PD-1H), especially in KBC pancreas Cancer model mice can significantly increase the tumor suppressor efficiency. It was found that the expression of PD-1H in T cells was significantly increased after administration of decitabine. Decitabine continued anti-PD-1H treatment could significantly prolong the survival of mice. [J]. Gastroenterology, 2017, 152 (5): S42-S43); WO2015035112 discloses an epigenetic modulator combined with an immunomodulator for the treatment of tumors, wherein the epigenetic modulator is selected from decitabine, immune The modulator can be selected from PD-1 / PD-L1 antibodies.

At present, clinical research has gradually carried out the treatment of decitabine combined with anti-PD-1 antibody, and the PD-1 antibody nivolumab (Nivolumab) combined with azacitidine in the treatment of relapsed and refractory AML (acute myeloid lymphocytic leukemia). The efficacy of 18% of patients in the study was evaluated as CR ([J] .Blood 2016 128: 763); a Phase I clinical study of PD-1 antibody and decitabine in advanced and untreated malignancies reported by Chunmeng Wang showed that Of the 11 patients enrolled One patient was evaluated as CR ([J]. Journal of Clinical Oncology 35 (2017): e14555-e14555). A Phase II clinical study of the PD-L1 inhibitor duvaluumab (Durvalumab) in combination with oral azacitidine in the treatment of myelodysplastic syndromes previously insensitive to decitabine or azacitidine (NCT02281084) ; PD-1 inhibitor Pembrolizumab combined with decitabine in the treatment of relapsed and refractory AML is also underway (NCT02996474), but the clinical results reported so far are still not ideal, and the overall survival of refractory tumors The prolongation of the period is not satisfactory, and the results of ongoing clinical studies are not yet known. How to screen the best PD-1 / PD-L1 inhibitor and DNA demethylation inhibitor to obtain the best tumor suppressive effect while taking into account Safety is still an urgent issue to be solved clinically.

The anti-PD-1 antibody provided by the present invention, WO2015085847 discloses the sequence and preparation method of the antibody. At present, the PD-1 antibody is in the domestic clinical phase I, and the safety is good. The reported clinical research results have shown that it has a certain Antitumor effect ([J]. Journal of Clinical Oncology 35 (2017): e15572-e15572).

The present invention provides the use of a combination of an immunotherapeutic agent and an epigenetic modulator in the preparation of a medicament for treating tumors and / or enhancing T-cells. The immunotherapeutic agent is selected from PD-1 antibodies.

The PD-1 antibody is known, and preferably the light chain variable region of the PD-1 antibody comprises 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 contains SEQ ID NO: 1, respectively. HCDR1, HCDR2 and HCDR3 shown in SEQ ID NO: 2 and SEQ ID NO: 3.

The CDR sequences described above are shown in the following table:

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

Preferably, the humanized antibody light chain variable region sequence is the sequence shown in SEQ ID NO: 10 or a variant thereof; the variant preferably has a 0 to 10 amino acid change in the light chain variable region. ; More preferably, 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; the variant preferably has a 0 to 10 amino acid change in the heavy chain variable region; more preferably G44R amino acid change.

The sequences of the variable regions of the heavy and light chains of the aforementioned humanized antibodies are as follows:

Heavy chain variable region SEQID NO: 9

Light chain variable region SEQID NO: 10

Preferably, the humanized antibody light chain sequence is the sequence shown in SEQ ID NO: 8 or a variant thereof; the variant preferably has a 0 to 10 amino acid change in the variable region of the light chain; more preferably A43S amino acid change. The humanized antibody heavy chain sequence is the sequence shown in SEQ ID NO: 7 or a variant thereof; the variant preferably has an amino acid change of 0 to 10 in the heavy chain variable region; more preferably G44R Amino acid changes.

The 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.

The sequences of the heavy and light chains of the aforementioned humanized antibodies are as follows:

Heavy chain SEQID NO: 7

Light chain SEQID NO: 8

In a preferred embodiment of the present invention, the epigenetic modulator is selected from the group consisting of a demethylating agent, a DNA methyltransferase (DNMT) inhibitor, a histone deacetylating enzyme (HDAC) inhibitor, or Multiple, wherein the DNMT inhibitor is selected from the group consisting of guadecitabine, RX-3117, lenalidomide, and azacitidine compound, EPI-01, decitabine, and E-7727 compound , RRx-001, temozolomide, CM-272, KM-101, KRX-0402, TdCyd, UVI-5008, azacitidine prodrug, decitabine prodrug, aza-T-dCyd, XB-05, PMX -700, CP-4200; the HDAC inhibitor is selected from the group consisting of mocetinostat, entinostat, purinostat, largazole, laxazole Gazolla analogue, ACY-738, resminostat, VRx-3996, givinostat, MPT-0E028, tucidinostat, TMB-ADC , Romidepsin, Par Panobinostat, 4SC-202, abexinostat, CUDC-907, vorinostat, pracinostat, HLY-0019, DWP- 0016, remetinostat, sodium phenylbutyrate, valproic acid, CKD-509, CKD-504, CKD-506, CKD-581, ricolinostat, CG- 200745, AR-42, SP-2-59, citarinostat, vorinostat, AP-001, quisinostat, SP-1-161, RCY -1497, belinostatin (belinostat), trichostatin A, TMP-195, RND-001, GSK-3117391, ACY-1083, tefinostat, tenosestat Tinostamustine, SF-2558HA, CXD-101, JW-1521, CG-1255, LB-205, LB-201, OCID-4681, QTX-153, APH-0812, CX-1026, OBP-801 , CS-3158, RG-2833, TL-112, HG-3001, KDAC-001, Thailandepsin B, QTX-125, RGFP-966, SP-1003, BEBT-908, BRD-3308, BEBT-201, BEBT-906, ACY-257, NMB-T-BMX-OS01, NBM-HD-1, zebularine.

In another preferred embodiment of the present invention, the epigenetic modulator is selected from the group consisting of azacitidine, decitabine, fludarabine, Guadecitabine, and zebralin. (Zebularine), NPEOC-DAC, CP-4200, Vorinostat, Romidepsin, Panobinostat, CI-994, 5,6-Dihydro-5- Azacitidine, 5-fluoro-2'-deoxycytidine, RX-3117, epigallocatechin gallate (EGCG), genistein, curcumin, oligonucleotides, preferably Selected from decitabine, azacitidine, Guadecitabine, rome Dimid (Romidepsin), Fludarabine (Fludarabine).

In a preferred embodiment of the present invention, the tumor is selected from malignant tumor and benign tumor; the malignant tumor is selected from malignant epithelial tumor, sarcoma, myeloma, leukemia, lymphoma, melanoma, head and neck tumor, brain tumor, Mixed tumor, malignant tumor in children; the malignant epithelial tumor is selected from the group consisting of lung cancer, breast cancer, liver cancer, pancreatic cancer, colorectal cancer, gastric cancer, gastroesophageal adenocarcinoma, esophageal cancer, small intestine cancer, cardiac cancer, endometrial cancer, ovary Cancer, fallopian tube cancer, vulvar cancer, testicular cancer, prostate cancer, penile cancer, kidney cancer, bladder cancer, anal cancer, gallbladder cancer, bile duct cancer, teratoma, heart tumor; the head and neck tumor is selected from nasopharyngeal cancer, throat 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 solitary myeloma Multiple myeloma, diffuse myeloma, leukemia myeloma, extramedullary myeloma; the leukemia is selected from the group consisting of acute lymphoblastic leukemia, chronic lymphocytic leukemia, Myeloid leukemia, chronic myeloid leukemia, hairy cell leukemia, T-cell lymphocytic leukemia, large granular lymphocytic leukemia, adult T-cell leukemia; the lymphoma is selected from non-Hodgkin's lymphoma, Hodgkin's lymphoma The brain tumor is selected from the group consisting of neuroepithelial tissue tumors, cranial nerve and spinal nerve tumors, and meningeal tissue tumors; the child malignant tumor is selected from nephroblastoma, neuroblastoma, retinoblastoma, and child germ cell tumor.

In another preferred embodiment of the present invention, the lung cancer is selected from the group consisting of non-small cell lung cancer and small cell lung cancer; the breast cancer is selected from the group consisting of hormone receptor (HR) positive breast Cancer, human epidermal growth Factor receptor-2 (HER2) positive breast cancer, triple negative breast cancer; the renal cancer is selected from the group consisting of clear renal cell carcinoma, papillary renal cell carcinoma, chromophobe renal cell carcinoma, and collecting duct cancer; the neuroepithelial tissue tumor Selected from the group consisting of preferred astrocytoma, anaplastic astrocytoma, and glioblastoma; the liver cancer is selected from primary liver cancer, secondary liver cancer, and the primary liver cancer is selected from hepatocellular carcinoma, bile duct cell carcinoma, Mixed liver cancer; the colorectal cancer is selected from colon cancer and rectal cancer.

In another preferred embodiment of the present invention, the tumor is selected from solid tumor, Hodgkin's lymphoma, non-Hodgkin's lymphoma, prostate cancer, pancreatic cancer, lung cancer, esophageal cancer, liver cancer, bile duct cancer, breast Cancer, colorectal cancer, gastric cancer, kidney cancer, acute myeloid lymphocytic leukemia, myelodysplastic syndrome, glioma.

In a preferred embodiment of the invention, the tumor is mediated and / or expresses PD-L1 by PD-1.

In a preferred embodiment of the present invention, the tumor is selected from intermediate and advanced tumors, relapsed refractory tumors, failed and / or relapsed tumors treated with chemotherapy drugs, failed and / or relapsed tumors treated with radiotherapy, and failed treatments with targeted drugs And / or relapsed tumor, failed immunotherapy, and / or relapsed tumor.

In a preferred embodiment of the present invention, the tumor is resistant or resistant to an immunotherapeutic agent or immunotherapy. Preferably, the immunotherapeutic agent is PD-1 and / or PD-L1 or CTLA- 4 (cytotoxic T lymphocyte-associated protein 4) as the target; the immunotherapy is selected from the group consisting of immunological checkpoint blocking (ICB) therapy, chimeric antigen receptor T cell immunotherapy (CAR-T therapy), and autologous cellular immunity Therapy (CIK therapy).

In a preferred embodiment of the present invention, preferably, the immunotherapy The agent is selected from the group consisting of PD-1 antibody, PD-L1 antibody, and CTLA-4 antibody. The PD-1 antibody includes, but is not limited to, pidilizumab, MEDI-0680, AMP-224, PF-06801591, and TSR- 042, JS-001, GLS-010, PDR-001, Genolimzumab, Camrelizumab, BGB-A317, IBI-308, REGN-2810, Pembrolizumab, Nivolumab; the PD-L1 antibody includes, but is not limited to, MSB-0011359-C, CA-170, LY-3300054, BMS-936559, Durvalumab, and Aveluzumab ( Avelumab), Atezolizumab; The CTLA-4 antibody includes, but is not limited to, ipilimumab, AK-104, JHL-1155, ATOR-1015, AGEN-1884, PRS-010, Trimelimumab, 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 use of a PD-1 antibody combined with an epigenetic regulator in the preparation of a T-cell activity-enhancing drug is provided. The T cells are preferably peripheral T-cells.

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

In a preferred embodiment of the present invention, the PD-1 antibody dose is selected from 50-600mg, preferably selected from 50mg, 60mg, 70mg, 75mg, 100mg, 125mg, 150mg, 175mg, 200mg, 225mg, 250mg, 375mg, 400mg, 425mg, 450mg, 475mg, 500mg, 600mg, more preferred From 100mg, 200mg, 400mg.

In a preferred embodiment of the present invention, the dose of the epigenetic modulator is selected from 5-100 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 ; more preferably selected from 7mg / m ² , 10mg / m ² , 12mg / m ² , 15mg / m ² , 20mg / m ² , 25mg / m ² , 30mg / m ² , 40mg / m ² , 50 mg / m ² , and 75 mg / m ² .

In a preferred embodiment of the present invention, the dose of the epigenetic modulator is selected from 5-500 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, more preferably selected from 10mg, 20mg, 100mg, 200mg, 300mg.

In a preferred embodiment of the present invention, the weight ratio of the PD-1 antibody to the epigenetic modulator is selected from 0.01 to 100: 1, preferably 5: 1, 4: 1, 3: 1, 5: 2, 2: 1, 7: 4, 3: 2, 4: 3, 5: 4, 1: 1, 3: 4, 2: 3, 3: 5, 1: 2, 2: 5, 1: 3, 3:10, 1: 4, 40: 1, 25: 1, 20: 1, 15: 1, 25: 2, 12: 1, 10: 1, 8: 1, 9: 2, 25: 4, 6: 1, 20: 3, 15: 4; more preferably selected from 5: 1, 4: 1, 3: 1, 5: 2, 2: 1, 1: 1: 1, 2: 3, 4: 3, 40: 1 20: 1.

In a preferred embodiment of the present invention, the combination further comprises a third component selected from the group consisting of alkylating agents, platinum complexing agents, metabolic antagonists, plant alkaloids, hormone anticancer agents, and proteasome inhibition. Agent, aromatase inhibitor, immunomodulator, EGFR inhibitor, ALK inhibitor, PARP inhibitor VEGF antibody, VEGFR inhibitor, mTOR inhibitor.

Preferably, the chemotherapeutic drug is selected from an alkylating agent, a platinum complexing agent, and a metabolic antagonist One or more of an antibiotic, a plant alkaloid (such as vinblastine, harringtonine), a hormonal anticancer agent, a proteasome inhibitor, an aromatase inhibitor, or an immunomodulator; in another preferred implementation In the example protocol, the chemotherapeutic drugs include, but are not limited to, cyclophosphamide, ifosfamide, melphalan, busulfan, nimustine, ramustine, dacarbazine, temozolomide, nitrogen mustard hydrochloride, Dibromomannitol, cisplatin, carboplatin, oxaliplatin, nedaplatin, methotrexate, 5-fluorouracil, tegafur, gemcitabine, capecitabine, fulvestrant, pemetrexed, Anthracycline antibiotics, mitomycin, bleomycins, actinomycins, vinblastine, camptothecin, paclitaxel, vincristine, vinblastine, vinblastine, etoposide, docetaxel Sytaxel, paclitaxel, albumin-bound paclitaxel, paclitaxel liposomes, irinotecan, vinorelbine, mitoxantrone, vinblastine, topotecan, leuprolide, goserelin, dutasteride, Fulvestrant, dexamethasone, tamoxifen, bortezomib, lenalidomide, etc., exemestane, Letrozole, Anastrozole.

Preferably, the targeted drug is selected from one or more of EGFR inhibitor, ALK inhibitor, PARP inhibitor, VEGF antibody and VEGFR inhibitor, mTOR inhibitor. These targeted drugs are well known in the art. For example, EGFR inhibitors can be selected from the group consisting of gefitinib, erlotinib, ektinib, and afatinib, cetuximab, trastuzumab One or more of ALK inhibitors; selected from crizotinib, ceritinib, axitinib, and brigatinib; VEGF antibodies selected from bevacizumab, bluzumab (Brolucizumab), Vanucizumab, Navicixizumab, Ranibizumab, Conbercept; VEGFR inhibitor selected from sunitinib, One or more of apatinib and famitinib; in a preferred embodiment of the present invention, the third component is selected from gemcitabine, cisplatin, carboplatin, paclitaxel albumin, paclitaxel liposomes, paclitaxel , One or more of docetaxel, cyclophosphamide, doxorubicin, epirubicin, vincristine, tigio, tegafur, 5-fluorouracil, and tetrahydrouridine.

In the above-mentioned preferred embodiment of the present invention, the above-mentioned third component may be based on the patient's body surface area, weight, or KPS functional status score standard or ECOG physical status score standard (Zubrod-ECOG-WHO) and various tumor diagnosis and treatment guidelines. In the selection of different types of tumor chemotherapy regimen recommended dosage and dosing regimen. For example, the dosage of paclitaxel albumin is 50 to 500 mg / m ² , preferably 125 mg / m ² ; the dosage of gemcitabine is 500 to 2000 mg / m ² , preferably 1000 mg / m ² ; the dosage of cisplatin is 25 To 200 mg / m ² , preferably 75 mg / m ² .

In the present invention, the treatment cycle may be 1 day, 3 days, 1 week, 2 weeks, 3 weeks, 3 to 4 weeks, 4 weeks, preferably 3 weeks or 3 to 4 weeks or 4 weeks.

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

In the present invention, the PD-1 antibody is used in combination with an epigenetic modulator for tumors, and the order of administration of the two is that the epigenetic modulator is administered before the PD-1 antibody is administered, or both are administered at the same time. Or the epigenetic modulator is administered after the PD-1 antibody is administered; preferably, the epigenetic modulator is administered before the PD-1 antibody is administered.

In the present invention, the epigenetic agent and the PD-1 antibody may be The same treatment cycle is used for the treatment of tumors. During the treatment of tumors, the epigenetic agent and the PD-1 antibody can be administered at the same time or before or after it can be combined according to the better chemotherapy regimen or radiation therapy of different tumors. Regimens or targeted small molecule drug treatment regimens or immunotherapy regimens to treat tumors, which include but are not limited to cellular immunotherapy (such as CAR-T therapy, tumor vaccine, CIK therapy, etc.); in addition, epigenetic agents and PD The combined administration of the -1 antibody can also be carried out alone in combination with other treatment regimens.

In the present invention, the PD-1 antibody and the epigenetic modulator can be used in combination with different tumor pathological types and advanced stages of tumor treatment schemes according to various tumor diagnosis and treatment guidelines or guidelines. Guiding principles include, but are not limited to, NCCN (National Comprehensive Cancer Network issued clinical practice guidelines for various malignancies) or the malignant tumor diagnosis and treatment guidelines issued by the Chinese Ministry of Health.

In the present invention, epigenetic modulators can be performed within 1 to 2 treatment cycles before traditional tumor treatment protocols (including but not limited to chemotherapy, radiotherapy, small molecule targeted therapy, surgical resection, and endoscopic treatment). Continuous administration over any period of 1 to 7 days, preferably continuous administration over a period of 1 to 5 days. During this treatment cycle, PD-1 antibody is administered simultaneously with the epigenetic modulator. Or before or after the epigenetic modulator, preferably, the PD-1 antibody can be administered on the 1st, 2nd, 3rd day after the end of the decitabine administration, Administration on the 4th, 5th, 6th, and 7th days, preferably on the 3rd, 4th, and 5th days; more preferably, PD-1 antibody can be administered on decitabine Dosing 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, and 7 days before starting, preferably 3 days, 4 days, and 5 days before starting; The epigenetic modulator is selected from decitabine and azacitidine; the dose of decitabine is preferably 10 mg or 7 mg / m ² .

The "combination" of the present invention is a mode of administration, which refers to the administration of at least one dose of PD-1 antibody and at least one dose of epigenetic modulators within a certain period of time, both of which exhibit pharmacological effects. The time period can be within one administration cycle, preferably within 4 weeks, within 3 weeks, within 2 weeks, within 1 week, or within 24 hours and within 12 hours. The PD-1 antibody and the epigenetic modulator can be administered simultaneously or sequentially. This term includes treatments in which PD-1 antibodies and epigenetic modulators are administered by the same route or different routes of administration. The combined mode of administration of the present invention is selected from simultaneous administration, independent formulation and co-administration, or independent formulation and sequential administration. In the present invention, the use of the medicament further related to the present invention, wherein the frequency of administration of the PD-1 antibody is once a day, twice a day, three times a day, once a week, once every two weeks, once every three weeks, one Once a month, the frequency of administration of epigenetic modulators 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, taking 21 days as a dosing cycle, decitabine is administered at a dose of 10 mg / day 5 days per week, and PD-1 antibody is administered at a dose of 4 mg / kg once a week; It can be administered in a fixed dose, 200mg or 400mg per patient (body weight greater than 80kg); this administration method can be circulated for 8 cycles.

In the most preferred embodiment, 21 days is a dosing cycle, and decitabine is administered at 10 mg / day on the 1st to 5th days of the first dosing cycle; on the 8th day The PD-1 antibody was administered at 4 mg / kg; the PD-1 antibody can also be administered at a fixed dose, 200 mg or 400 mg per patient (those weighing more than 80 kg). In a preferred embodiment of the present invention, the PD-1 antibody is administered by injection, such as subcutaneously or intravenously, and the PD-1 antibody needs to be formulated into an injectable form before injection. A particularly preferred injectable form of the PD-1 antibody is an injection solution or a lyophilized powder needle, which contains the PD-1 antibody, a buffer, a stabilizer, and optionally a surfactant. The buffer may be selected from one or more of acetate, citrate, succinate, and phosphate. The stabilizer may be selected from sugars or amino acids, preferably disaccharides such as sucrose, lactose, trehalose, maltose. The surfactant is selected from the group consisting of 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 vitamin polysorbate 20. The best injectable forms of PD-1 antibodies include PD-1 antibodies, acetate buffers, 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 the above-mentioned immunotherapeutic agent in combination with the above-mentioned epigenetic modulator as a medicine for treating and preparing tumors and / or enhancing T-cell activity.

In the present invention, a method for treating tumors and / or enhancing T-cell activity is provided, which comprises administering the above-mentioned immunotherapeutic agent and the above-mentioned epigenetic modulator to a patient.

The present invention also provides a medicine kit or a medicine package containing the aforementioned epigenetic modulator and PD-1 antibody.

The present invention also provides a medicinal composition comprising the aforementioned effective amount of PD-1 antibody and epigenetic modulator, and one or more pharmaceutically acceptable excipients, diluents or carriers.

Figure 1 is a comparison of the efficacy of drug resistance of PD-1 antibody combined with decitabine and PD-1 antibody administration.

Figure 2 shows the tumor regression after 4 weeks of PD-1 antibody combined with decitabine.

Figure 3 shows the reduction of tumor burden of malignant solid tumors with PD-1 antibody combined with decitabine.

Figure 4 shows the change of tumor burden of PD-1 antibody combined with decitabine in the treatment of malignant solid tumors with treatment time.

Detailed description of the invention

I. Terminology

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 "immunotherapy" refers to the use of 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 increasing the immunogenicity of tumor cells and the sensitivity to killing of effector cells, and Immune cells and effector molecules are infused into the host to cooperate 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 implementations The examples do not limit the scope of the invention.

Example 1: Treatment of relapsed and refractory Hodgkin lymphoma with decitabine combined with PD-1 antibody

1.Test antibodies and compounds

The PD-1 antibody is prepared according to the method disclosed in WO2015085847, and its corresponding code is H005-1. The sequences of its heavy and light chains are as SEQ ID NO: 7 and SEQ ID NO: 8 in the present invention. Lot number: P1512, 200mg / bottle, formulated as 20mg / ml for future use.

Commercially available decitabine lyophilized powder injection, 50mg / bottle, reconstituted with sterile water for injection, formulated into a solution of about 5mg / mL, and then formulated with physiological saline, 0.5% glucose solution or lactic acid-green solution to 0.1 according to clinical needs Use a -1 mg / mL solution.

Participants

(1) Hodgkin's lymphoma pathological diagnosis; (2) Ages 12 to 80 years; (3) ECOG score less than 2 points; (4) Life expectancy of at least 3 months; (5) Lymphoma subjects with lymphoma The tumor response standard defines at least one measurable lesion of 1 cm; (6) must have received at least two chemotherapy and have undergone an elution period of at least 4 weeks; the subject of autologous hematopoietic stem cell transplantation is eligible for more than 3 months; ( 7) Must have sufficient bone marrow, liver, kidney, lung and heart functions.

3.Grouping

As of September 27, 2017, a total of 19 subjects were enrolled in the study. Among them, 10 were resistant to PD-1 antibody treatment and were grouped into a resistance group treated with PD-1 antibody; 9 were not treated with PD-1 antibody and grouped to a group without PD-1 antibody treatment; the final effective evaluation targets were: There were 9 PD-1 antibody-resistant groups and 9 non-PD-1 antibody-treated groups. Among the subjects in the PD-1 antibody resistance group, 8 subjects had previously received 3 to 8 treatments with nivolumab, and 1 subject had previously received nivolumab at the same time (Nivolumab) and Pembrolizumab; 3 subjects in the non-PD-1 antibody-treated group had previously received CAR-T therapy; 2 of the 18 patients who were ultimately evaluated for efficacy He received both nivolumab and CAR-T.

4. Method of administration

Enrolled subjects were given decitabine 10 mg / d on the 1st to 5th days of the first dosing cycle (day 21), intravenous infusion; 200 mg of PD-1 antibody was administered on the 8th day (if the body weight exceeds 80 kg, 400 mg was administered), and this administration method was repeated for 8 cycles.

5. Test results

The results of clinical studies (see Table 1) show that during the clinical study, no serious adverse reactions occurred. The most common adverse reactions were cherry-like hemangiomas, fever, nausea, and rashes, all of which were grade 1 to 2 adverse reactions; decistat The total remission rate of Bin + PD-1 antibody in the treatment of relapsed and refractory Hodgkin's lymphoma was as high as 78%, and the objective remission rate was still as high as 56% in the drug-resistant group treated with PD-1 antibody. 1 / PD-L1 provides a new treatment approach for target-resistant patients.

Note: The drug resistance group treated with PD-1 antibody refers to the treatment with PD-1 antibody Four weeks without objective response (CR + PR); disease recurrence after PD-1 antibody treatment.

At the end of the experiment, a total of 71 patients were enrolled, of which one group was treated with PD-1 antibody alone. The specific results are as follows:

Example 2: Treatment of relapsed and refractory malignant tumors with decitabine combined with PD-1 antibody

1.Test antibodies and compounds

See Example 1

Participants

(1) Pathological diagnosis of malignant tumors; (2) Ages 12 to 80 years; (3) ECOG score less than 2 points; (4) Life expectancy of at least 3 months; (5) At least one 1 cm Measurable lesions; (6) Must fail first-line chemotherapy and have experienced an elution period of at least 4 weeks; (7) Must have sufficient bone marrow, liver, kidney, lung, and heart functions.

3.Grouping

As of October 30, 2016, a total of 11 subjects were enrolled, including 6 diffuse large B-cell lymphomas, 1 B-cell lymphoma, 1 Hodgkin lymphoma, and 4 solid tumors ( 2 gastric cancers, 1 esophageal cancer); the final evaluable object was 9 people. Two of the subjects with diffuse large B-cell lymphoma had been treated with CAR-T, and one subject with gastric cancer had previously been treated with CIK.

4. Method of administration

Enrolled subjects were given decitabine 10 mg / d on the 1st to 5th days of the first dosing cycle (day 21), intravenous drip; PD-1 antibody was given on the 8th day 4mg / kg, 8 cycles of administration were circulated according to this administration method.

5. Test results

The results of clinical studies (see Table 2) show that the total response rate of decitabine + PD-1 antibody in the treatment of relapsed and refractory malignant tumors was 7/9 (78%), of which 4 subjects received the PD-1 antibody monotherapy After the progress of treatment, PD-1 antibody combined with decitabine treatment has achieved a complete or partial response evaluation, which undoubtedly provides a new option for extending the survival of this type of patients.

Example 3: Treatment of relapsed and refractory malignant tumor with decitabine combined with PD-1 antibody

1.Test antibodies and compounds

See Example 1

Participants

(1) Pathological diagnosis of malignant tumors; (2) Ages 12 to 80 years; (3) ECOG score less than 2 points; (4) Life expectancy of at least 3 months; (5) At least one 1 cm Measurable lesions; (6) Must fail first-line chemotherapy and have experienced an elution period of at least 4 weeks; (7) Must have sufficient bone marrow, liver, kidney, lung, and heart functions.

3.Grouping

As of September 26, 2017, a total of 24 subjects were enrolled, including 18 Hodgkin lymphomas, 2 B-cell non-Hodgkin lymphomas; 4 solid tumors (2 colorectal cancer, 1 kidney cancer, 1 breast cancer); the final evaluable object was 23 people. Eight of the 18 Hodgkin's lymphoma subjects had previously received 3 to 13 treatments with nivolumab or Pembrolizumab, and 5 had previously received CAR-T therapy; 2 had B One of the non-Hodgkin's lymphoma subjects had previously received 13 nivolumab treatments, one had previously received CAR-T therapy, and two of the four solid tumor subjects had previously received 2 Or 10 times navumab (Nivolumab) treatment, 1 patient received 4 previous CIK treatments.

4. Method of administration

Enrolled subjects were given decitabine 10 mg / d on the 1st to 5th days of the first dosing cycle (day 21) and given an IV infusion; PD-1 antibody was given 4 mg / kg on the 8th day, and then given The medicine method was cycled for 8 dosing cycles.

5. Test results

The results of clinical studies show (see Figures 3 and 4) that decitabine + PD-1 antibody in the treatment of relapsed and refractory malignant tumors in accordance with the anti-tumor cell immunotherapy efficacy evaluation criteria (imRC-standard). The response rate was 10/23 (44%), of which 5 subjects had a complete response (CR, the tumor burden decreased by more than 100% compared to the baseline level), and 5 subjects had a partial response (PR, compared to the baseline level) Specific tumor load decreased by more than 50%), 10 subjects had stable disease (SD, tumor load decreased by less than 25%, but did not increase by more than 25% compared to baseline levels), and 2 subjects had disease progression (PD, and (Baseline level increased tumor load by more than 25%). According to the irRC standard, even if the patient's curative effect is evaluated as SD, it is actually considered to be a clinically beneficial population. Therefore, the rate of disease control (DCR) of decitabine + PD-1 antibody for relapsed and refractory malignant tumors is 21/23 (91% ). With the examples of the present invention, the treatment of relapsed and refractory malignant tumors with decitabine + PD-1 antibody is undoubtedly prolonging the survival of patients who have tolerated or relapsed with PD-1 antibody therapy and who have tolerated or relapsed with CAR-T or CIK therapy. Phase provides an important treatment path.

<110> Jiangsu Hengrui Pharmaceutical Co., Ltd. General Hospital of Chinese People's Liberation Army

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Claims (24)

The use of an immunotherapeutic agent and an epigenetic modulator in the preparation of a medicament for treating tumors and / or enhancing T-cell activity is characterized in that the immunotherapeutic agent is selected from the group consisting of PD-1 antibody, The light chain variable region contains 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 contains as shown in SEQ ID NO: 1, HCDR1, HCDR2 and HCDR3 shown in SEQ ID NO: 2 and SEQ ID NO: 3. The use according to item 1 of the patent application scope, wherein the PD-1 antibody is a humanized antibody. The use according to item 2 of the scope of patent application, wherein the light chain variable region sequence of the anti-PD-1 humanized antibody is the sequence shown in SEQ ID NO: 10 or a variant thereof; the heavy chain is variable The region sequence is the sequence shown in SEQ ID NO: 9 or a variant thereof. The use according to item 3 of the scope of patent application, wherein the variant has a 0 to 10 amino acid change in the variable region of the light chain. The use according to item 4 of the scope of patent application, wherein the variant is an amino acid change of A43S in the variable region of the light chain. The use as described in claim 3 of the patent application range, wherein the variant has a 0 to 10 amino acid change in the heavy chain variable region. The use as described in claim 6 of the scope of patent application, wherein the variant is an amino acid change of G44R in the variable region of the heavy chain. The use according to any one of claims 1 to 7, wherein the humanized antibody light chain sequence is the sequence shown in SEQ ID NO: 8, The heavy chain sequence is the sequence shown in SEQ ID NO: 7. The use according to any one of claims 1 to 8, wherein the epigenetic regulator is selected from the group consisting of a demethylating agent, a DNA methyltransferase (DNMT) inhibitor, and a histone deacetylamidine One or more inhibitors of HDAC. The use according to any one of claims 1 to 9, wherein the epigenetic modulator is selected from the group consisting of azacitidine, decitabine, fludarabine, and orodicitabine Guadecitabine, Zebularine, NPEOC-DAC, CP-4200, Vorinostat, Romidepsin, Panobinostat, CI-994 , 5,6-dihydro-5-azacytidine, 5-fluoro-2'-deoxycytidine, RX-3117, epigallocatechin gallate (EGCG), genistein, Curcumin, oligonucleotides. The use according to item 10 of the scope of patent application, wherein the epigenetic modulator is selected from the group consisting of decitabine, azacitidine, guancitabine, romidepsin and fludarabine. The use according to any one of claims 1 to 11, wherein the tumor is selected from malignant tumors and benign tumors; the malignant tumor is selected from malignant epithelial tumors, sarcoma, myeloma, leukemia, lymphoma, melanin Tumors, head and neck tumors, brain tumors, mixed tumors, childhood malignancies. The use according to any one of claims 1 to 12, wherein the tumor is selected from solid tumors, Hodgkin's lymphoma, non-Hodgkin's lymphoma, prostate cancer, pancreatic cancer, lung cancer, and esophageal cancer , Liver cancer, bile duct cancer, Breast cancer, colorectal cancer, gastric cancer, kidney cancer, acute myeloid lymphocytic leukemia, myelodysplastic syndrome, glioma. The use according to any one of claims 1 to 13, wherein the tumor is mediated by PD-1 and / or expresses PD-L1. The use according to any one of claims 1 to 14, in which the tumor is selected from the group consisting of intermediate and advanced tumors, relapsed refractory tumors, failed and / or relapsed tumors treated with chemotherapy drugs, failed radiation therapy and / or Relapsed tumors, failed and / or relapsed tumors after targeted drug treatment, failed and / or relapsed tumors after immunotherapy. The use according to any one of claims 1 to 15, wherein the tumor is resistant or resistant to an immunotherapeutic agent or immunotherapy. The use according to any one of claims 1 to 16, wherein the tumor is resistant or resistant to an immunotherapeutic agent that targets PD-1 or PD-L1 or CTLA-4. The use according to any one of claims 1 to 17, wherein the dose of the PD-1 antibody is selected from 1 to 10 mg / kg. The use according to item 18 of the scope of patent application, wherein the PD-1 antibody dose is selected from 3 mg / kg, 4 mg / kg, and 5 mg / kg. The use according to any one of claims 1 to 19 of the application, wherein the PD-1 antibody dose is selected from 50 to 600 mg, wherein the dose is administered every three weeks. The use according to item 20 of the patent application range, wherein the PD-1 antibody dose is selected from 200 mg and 400 mg each time. The use according to any one of claims 1 to 21, wherein the epigenetic modulator is administered at a dose of 10 mg / day, of which 5 days are given every 21 days. A pharmaceutical packaging box, comprising the epigenetic regulator and PD-1 antibody according to any one of claims 1 to 22 of the scope of patent application. A medicinal composition, characterized in that it comprises an effective amount of PD-1 antibody and an epigenetic regulator as described in any one of claims 1 to 22 of the scope of patent application, and one or more pharmaceutically acceptable excipients Agent, diluent or carrier.