CN109663130A

CN109663130A - PD-1 antibody and mek inhibitor combine the purposes in the drug of preparation treatment tumour

Info

Publication number: CN109663130A
Application number: CN201811186529.1A
Authority: CN
Inventors: 邹建军; 徐瑞华; 刘毅
Original assignee: Jiangsu Hengrui Medicine Co Ltd
Current assignee: Jiangsu Hengrui Medicine Co Ltd
Priority date: 2017-10-13
Filing date: 2018-10-12
Publication date: 2019-04-23
Anticipated expiration: 2038-10-12
Also published as: CN109663130B

Abstract

The present invention relates to PD-1 antibody or its antigen-binding fragment and mek inhibitor to combine the purposes in the drug of preparation treatment tumour.Specifically, the present invention relates to PD-1 antibody or its antigen-binding fragments and compound (1) or its officinal salt to combine the purposes in the drug of preparation treatment tumour.

Description

PD-1 antibody and mek inhibitor combine the purposes in the drug of preparation treatment tumour

Technical field

PD-1 antibody or its antigen-binding fragment and mek inhibitor combine the purposes in the drug of preparation treatment tumour.

Background technique

The occurrence and development of malignant tumour and the aberrant continuation activation of Cellular Signaling Transduction Mediated access are closely related.MAPK The signal path control of (Mitogen-activated protein kinase, the protein kinase of mitosis person's activation) composition Multiple important physiology courses of cell, wherein ERK access (Extracellular regulated protein kinase Pathway, RAS-RAF-MEK1/2-ERK1/2, extracellular regulated protein kinases) it is the cascade signal occurred most frequently in human tumor Abnormal classical MAPK access.The abnormal activation of ERK access is usually by RAS (Rat sarcoma gene, rat sarcoma albumen Homologous gene) and RAF (Rapidly Accelerated Fibrosarcoma, rapid type accelerating fibers sarcoma kinases) gene man The gain mutation of family member leads to ([J] .Biochimica et Biophysica Acta (BBA)-Molecular Cell Research,2007,1773(8):1263-1284.).Wherein cholangiocarcinoma, colorectal cancer, lung cancer, bladder cancer, cancer of pancreas and forefront There is RAS encoding gene in gland cancer, and there are BRAF genes to dash forward for melanoma, papillary thyroid carcinoma and low differentiation oophoroma Become.BRAF and its downstream targets MEK is the kinases in MAPK access, and play an important role ([J] .Drug in cell Proliferation design,development and therapy,2016,10:43.).And PD-1 (programmed death receptor 1) antibody can be special Opposite sex identification and bind lymphocytes surface PD-1 block PD-1/PD-L1 signal path, and then T cell are activated to exempt from tumour Epidemic disease lethal effect transfers body immune system and removes interior tumor cell.

With gradually going deep into for PD-1/PD-L1 immunization therapy research, it is found that it is unwise many patients treat immunologic test point Sense or therewith drug resistance, the meta-analysis of Xia Bu et al. is studies have shown that can by the immunization therapy drug resistance of target spot of PD-1/PD-L1 Can be related with the up-regulation of certain genes, following such drag-resistance marker such as IL-10, VEGFA, VEGFC, FLT1, ANGPT2 and Monocyte and the macrophage driveization factor ([J] .Trends in molecular medicine, 2016,22 (6): 448- 451.) Immune expression is participated in.MEK protein kinase be normal T-cell and RAS signal access mutated tumor important regulating and controlling target spot it One, inhibit MEK that can block the proliferation of mouse Naive T cells, and pass through the front activating of the Apoptosis enhancing of damage TCR driving The anti-tumor activity of T cell, therefore mek inhibitor and can produce lasting tumour suppression with the combination of the antibody of PD-1/PD-L1 target spot Effect ([J] .Immunity, 2016,44 (3): 609-621.) processed.WO2016040892 is disclosed selected from Pembrolizumab Deng PD-1 antibody and mek inhibitor combination treatment kinds of tumors；WO2016011160 disclose selected from Nivolumab, A variety of B-RAF antagonist tolerance tumours are treated in PD-1 antibody and the mek inhibitor combination of Pembrolizumab etc.； WO2014195852 and WO2014193898 discloses Trametinib or Dabrafenib joint and PD-L1 antibody or PD-1 are anti- Body combination treatment B-RAF V600 saltant type melanoma；WO2013019906 discloses a variety of PD-1 antibody and a variety of MEK press down Agents treat B-RAF V600E mutant tumours.

Have multinomial PD-1 or PD-L1 joint mek inhibitor at present carrying out for treating the clinical research of tumour. Wilson H.Miller et al. open Atezolizumab (PD-L1) joint cobimetinib treatment metastasis melanin tumor Clinical studies show, the two combination compared with the two is used alone there is higher DCR (disease control rate) He Zhongwei to get nowhere Life cycle (mPFS) (Atezolizumab (A)+cobimetinib (C) in metastatic melanoma (mel): Updated safety and clinical activity[J].2017.)；Michelle Rohlfs et al. reports BRAF suppression The +/- mek inhibitor of preparation and combine PD-1 immunologic test order blocking agent (pembrolizumab or nivolumab) treatment transfer Property melanoma clinical study results, display patient in group mPFS is 7.6 months, wherein BRAF inhibitor+mek inhibitor+PD- 1 immunologic test order blocking agent group be 8.5 months, BRAF inhibitor+PD-1 immunologic test order blocking agent group be 7.2 months, it is tested Person's well-tolerated (BRAF with or without MEK inhibition plus PD-1 checkpoint blockade for the treatment of metastatic melanoma[J].2016.)；Antoni Ribas et al. report Pembrolizumab combines dabrafenib and trametinib and treats BRAF^V600E/KThe clinic of the advanced melanoma of mutation Result of study shows that the ORR (Overall response rate) in evaluable 15 subjects is 60%, but neutral grain occurs for 3 subjects The serious adverse reactions such as Leukopenia (Pembrolizumab in combination with dabrafenib and trametinib for BRAF-mutant advanced melanoma:Phase 1KEYNOTE-022 study[J] .2016)。

In conclusion although the research about PD-1 antibody and mek inhibitor with BRAF it has been reported that but mostly inhibited at present Agent is combined based on mek inhibitor and PD-L1 antibody, and subject's clinical Benefit and safety are not up to satisfactory effect, Therefore there is still a need for further explore suitable PD-1 antibody and mek inhibitor combination treatment recurrent and refractory tumour for therapy field.

Anti- PD-1 antibody provided by the invention, WO201508584 disclose the sequence and preparation method of the antibody, at present PD-1 antibody is in domestic clinical I phase, good security, and it is certain that the clinical study results registered have shown that it has Antitumor action ([J] .Journal of Clinical Oncology 35 (2017): e15572-e15572)；In addition originally The mek inhibitor provided is invented, WO2015058589 discloses its structure and preparation method, and in vitro study, which is shown, to be had preferably Tumors inhibition activity, specific structure are as follows:

Summary of the invention

The present invention provides a kind of immunotherapeutic agent and mek inhibitor combines the purposes in the drug of preparation treatment tumour, The immunotherapeutic agent is selected from PD-1 antibody or its antigen-binding fragment；The mek inhibitor is selected from AZD-8330, GDC- 0623、CI-1040、WX-554、TAK-733、HL-085、BI-847325、CEP-1347、Binimetinib、Pimasertib、 Cobimetinib、PD-0325901、RO-5126766、Trametinib、Refametinib、AS-703988、E-6201、 Selumetinib or compound as follows (1) or its officinal salt,

In preferred embodiment of the invention scheme, the mek inhibitor preferably be selected from Binimetinib, Pimasertib, Cobimetinib, Trametinib, Refametinib, compound (1) or its officinal salt.

PD-1 antibody or its antigen-binding fragment are known, preferably described PD-1 antibody or its antigen-binding fragment Light chain variable region include respectively LCDR1, LCDR2 as shown in SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6 and LCDR3。

The heavy chain variable region of the PD-1 antibody or its antigen-binding fragment includes respectively such as SEQ ID NO:1, SEQ HCDR1, HCDR2 and HCDR3 shown in ID NO:2 and SEQ ID NO:3.

Wherein, mentioned-above each CDR sequence is as shown in the table:

Title	Sequence	Number
			HCDR1	SYMMS	SEQID NO:1
HCDR2	TISGGGANTYYPDSVKG	SEQID NO:2
			HCDR3	QLYYFDY	SEQID NO:3
LCDR1	LASQTIGTWLT	SEQID NO:4
			LCDR2	TATSLAD	SEQID NO:5
LCDR3	QQVYSIPWT	SEQID NO:6

Preferably, the PD-1 antibody or its antigen-binding fragment are PD-1 humanized antibody.

Preferably, humanized antibody light chain's variable region sequences are the sequence as shown in SEQ ID NO:10 or its variant；Institute The variant stated preferably has the amino acid of 0-10 to change in light chain variable region；The amino acid of more preferably A43S changes.The source of people Changing antibody heavy chain variable region sequence is the sequence as shown in SEQ ID NO:9 or its variant；The variant is preferably in weight chain variable Area has the amino acid of 0-10 to change；The amino acid of more preferably G44R changes.

Humanized antibody above-mentioned is heavy, the variable region sequences of light chain are as follows:

Heavy chain variable region

EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYMMSWVRQAPGKGLEWVATISGGGANTYYPDSVKGRFT ISRDNAKNSLYLQMNSLRAEDTAVYYCARQLYYFDYWGQGTTVTVSS

SEQID NO:9

Light chain variable region

DIQMTQSPSSLSASVGDRVTITCLASQTIGTWLTWYQQKPGKAPKLLIYTATSLADGVPSRFSGSGSGT DFTLTISSLQPEDFATYYCQQVYSIPWTFGGGTKVEIK

SEQID NO:10

Preferably, humanized antibody light chain's sequence is the sequence as shown in SEQ ID NO:8 or its variant；The variant It is preferred that thering is the amino acid of 0-10 to change in light chain variable region；The amino acid of more preferably A43S changes.The humanized antibody weight Chain-ordering is the sequence as shown in SEQ ID NO:7 or its variant；The variant preferably has the amino of 0-10 in heavy chain variable region Acid variation；The amino acid of more preferably G44R changes.

Particularly preferred PD-1 humanized antibody light chain's sequence is the sequence as shown in SEQ ID NO:8, heavy chain Sequence is the sequence as shown in SEQ ID NO:7.

PD-1 humanized antibody above-mentioned is heavy, the sequence of light chain is as follows:

Heavy chain

EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYMMSWVRQAPGKGLEWVATISGGGANTYYPDSVKGRFT ISRDNAKNSLYLQMNSLRAEDTAVYYCARQLYYFDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVK DYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCP PCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVS VLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK

SEQID NO:7

Light chain

DIQMTQSPSSLSASVGDRVTITCLASQTIGTWLTWYQQKPGKAPKLLIYTATSLADGVPSRFSGSGSGT DFTLTISSLQPEDFATYYCQQVYSIPWTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKV QWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

SEQID NO:8

In preferred embodiment of the invention scheme, in preferred embodiment of the invention scheme, the tumour is selected from pernicious Tumour, benign tumour；The malignant tumour is selected from carcinoma, sarcoma, myeloma, leukaemia, lymthoma, melanin Tumor, head and neck neoplasm, brain tumor, peritoneal cancer, mixed type tumour, children malignant tumors；The carcinoma is selected from lung Cancer, breast cancer, liver cancer, cancer of pancreas, colorectal cancer, gastric cancer, stomach oesophagus gland cancer, the cancer of the esophagus, carcinoma of small intestine, cardia cancer, endometrium Cancer, oophoroma, carcinoma of fallopian tube, carcinoma of vulva, carcinoma of testis, prostate cancer, carcinoma of penis, kidney, bladder cancer, cancer of anus, gallbladder cancer, gallbladder Pipe cancer, teratoma, cardiac tumor；The head and neck neoplasm is selected from nasopharyngeal carcinoma, laryngocarcinoma, thyroid cancer, tongue cancer, carcinoma of mouth；It is described Sarcoma is selected from Askin's tumor, chondrosarcoma, ewing's sarcoma, malignant hemangioma, malignant schwannoma, osteosarcoma, soft tissue Sarcoma；The myeloma is selected from stand alone myeloma, multiple types myeloma, diffuses type myeloma, leukemia-type myeloma, outside marrow Type myeloma；The leukaemia is selected from acute lymphatic leukemia, chronic lymphatic leukemia, acute myelogenous leukemia, slow Property myelomatosis, hairy cell leukaemia, T cell lymphocytic leukemia, large granular lymphocyte leukaemia, at Human T cell leukemia；The lymthoma is selected from non-Hodgkin lymphoma, Hodgkin lymphoma；The brain tumor is selected from nerve Epithelial neoplasms, cranial nerve and spinal nerve tumour, meningeal tissue tumour；The children malignant tumors are selected from kidney mother cell Tumor, neuroblastoma, retinoblastoma, children's germinoma.

In another preferred embodiment scheme of the invention, the lung cancer is selected from the lung cancer and is selected from non-small cell lung Cancer, Small Cell Lung Cancer；The breast cancer is selected from the breast cancer and is selected from hormone receptor (HR) positive breast cancer, human epidermal growth factor Sub- receptor -2 (HER2) positive breast cancer, three negative breast cancer；The kidney be selected from transparent clear-cell carcinoma, Papillary Renal Cell Carcinoma, Chromophobe property clear-cell carcinoma, Collecting duct carcinoma；The neuroepithelial tissue tumour is selected from preferred astrocytoma, anaplastic astrocytoma Cytoma, glioblastoma；The liver cancer is selected from primary carcinoma of liver, secondary carcinoma of liver, and it is thin that the primary carcinoma of liver is selected from liver Born of the same parents' cancer, cholangiocellular carcinoma, Combination liver cancer；The colorectal cancer is selected from colon and rectum carcinoma.

In another preferred embodiment scheme of the invention, the tumour is selected from Hodgkin lymphoma, non-Hodgkin's It is lymthoma, prostate cancer, cancer of pancreas, lung cancer, the cancer of the esophagus, liver cancer, cholangiocarcinoma, breast cancer, colorectal cancer, gastric cancer, kidney, acute The unknown tumour of marrow lymphocytic leukemia, myelodysplastic syndrome, glioma, original site.

In preferred embodiment of the invention scheme, the tumour mediates and/or expresses PD-L1 by PD-1.

In preferred embodiment of the invention scheme, the tumour be selected from RAS mutant tumours, RAF mutant tumours, RAS/RAF/MEK access ectype tumour, RAS/RAF access ectype tumour.

In preferred embodiment of the invention scheme, the RAS saltant type be selected from HRas saltant type, KRas saltant type, NRas saltant type；The RAF saltant type is selected from A-RAF saltant type, B-RAF saltant type.Wherein B-RAF saltant type preferably is selected from B- RAF V600E saltant type, B-RAF V600K saltant type, B-RAF V600D saltant type, B-RAF V600R saltant type.

In the present invention, above-mentioned to sport Positive mutants.

In preferred embodiment of the invention scheme, the tumour is selected from Advanced cancers, recurrent and refractory tumour, through changing Treat drug therapy failure and/or recurrent tumor, through radiotherapy failure and/or recurrent tumor, through targeted drug treatment failure and/or multiple Swell tumor, through immunization therapy failure and/or recurrent tumor.

In preferred embodiment of the invention scheme, the tumour is to immunotherapeutic agent or immunotherapy or shows as resisting Or drug resistance, it is preferred that the immunotherapeutic agent is that (cytotoxic T lymphocyte is related with PD-1 and/or PD-L1 or CTLA-4 Albumen 4) it is target spot；The immunotherapy is selected from immunologic test point and blocks (ICB) therapy, Chimeric antigen receptor T cell are immune to treat Method (CAR-T therapy), autogenous cell immunotherapy (CIK therapy).

In preferred embodiment of the invention scheme, it is preferred that it is anti-that the immunotherapeutic agent is selected from PD-1 antibody, PD-L1 Body, CTLA-4 antibody, the PD-1 antibody includes but is not limited to 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 antibody includes but is not limited to MSB- 0011359-C,CA-170,LY-3300054,BMS-936559,Durvalumab,Avelumab,Atezolizumab；It is described CTLA-4 antibody include but is 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 preferred embodiment of the invention scheme, the PD-1 antibody or its antigen-binding fragment dosage are selected from 1- 10mg/kg, preferably be selected from 1mg/kg, 2mg/kg, 3mg/kg, 4mg/kg, 5mg/kg, 6mg/kg, 7mg/kg, 8mg/kg, 9mg/kg, 10mg/kg, more preferable 1mg/kg, 3mg/kg, 4mg/kg, 5mg/kg, 6mg/kg, 10mg/kg.

In preferred embodiment of the invention scheme, the PD-1 antibody or its antigen-binding fragment dosage are selected from 50- 600mg, preferably be selected from 50mg, 60mg, 70mg, 75mg, 100mg, 125mg, 150mg, 175mg, 200mg, 225mg, 250mg, 375mg, 400mg, 425mg, 450mg, 475mg, 500mg, 600mg, more preferably from 60mg, 100mg, 200mg, 400mg, 600mg。

In preferred embodiment of the invention scheme, the mek inhibitor dosage be selected from 0.01-500mg, preferably be selected from 0.1mg, 0.25mg、0.5mg、0.75mg、1mg、2mg、3mg、4mg、5mg、6mg、7mg、8mg、9mg、10mg、11mg、12mg、 12.5mg、15mg、17.5mg、20mg、22.5mg、25mg、30mg、45mg、50mg、60mg、70mg、75mg、80mg、90mg、 100mg, 125mg, 150mg, 175mg, 200mg, 225mg, 250mg, 275mg, 300mg, 400mg, 500mg, more preferably 0.25mg、0.5mg、1mg、2mg、3mg、4mg、10mg、15mg、20mg、30mg、45mg、50mg、60mg、75mg、100mg。

In preferred embodiment of the invention scheme, the mek inhibitor is selected from compound (1) or its officinal salt, agent Amount be selected from 0.01-500mg, preferably be selected from 0.1mg, 0.25mg, 0.5mg, 0.75mg, 1mg, 2mg, 3mg, 4mg, 5mg, 6mg, 7mg, 8mg、9mg、10mg、11mg、12mg、12.5mg、15mg、17.5mg、20mg、22.5mg、25mg、30mg。

In preferred embodiment of the invention scheme, the officinal salt of the compound (1) is selected from tosilate.

In the present invention, PD-1 antibody or its antigen-binding fragment are combined with mek inhibitor for oncotherapy, the two Be administered order be mek inhibitor be administered PD-1 antibody or its antigen-binding fragment administration before, or both be administered simultaneously, or Mek inhibitor administration is after PD-1 antibody or the administration of its antigen-binding fragment；Preferably, mek inhibitor administration is anti-in PD-1 Before body or the administration of its antigen-binding fragment.

In the present invention, PD-1 antibody or its antigen-binding fragment are combined with mek inhibitor for oncotherapy, and the two can It is administered in the same dosage period.

In the present invention, the treatment cycle can be 1 day, 3 days, 1 week, 2 weeks, 3 weeks (21 days), 3-4 weeks (21-28 days), 4 All (28 days), preferably 3 weeks or 3-4 weeks or 4 weeks.

In the present invention, the treatment cycle includes but is not limited to chemotherapy cycles or radiotherapy period or other correlation targeting medicines Object treatment cycle or immunization therapy period.

In the present invention, mek inhibitor and PD-1 antibody or its antigen-binding fragment can be in identical or different treatments Period is inline to be shared in treatment tumour, during treat tumour, mek inhibitor and PD-1 antibody or its antigen-binding fragment While administering drug combinations or before or after can also combine according to the preferred chemotherapy regimen of different tumours or radiotherapy in the treatment scheme or It targets small molecule treatment scheme or Immunotherapy regimens treats tumour, the Immunotherapy regimens include but is not limited to cell Immunotherapy (such as CAR-T therapy, tumor vaccine, CIK therapy etc.)；Furthermore mek inhibitor and PD-1 antibody or its antigen binding The administering drug combinations of segment, which can not also combine other treatment schemes, individually to carry out.

In the present invention, PD-1 antibody or its antigen-binding fragment and mek inhibitor be while combination, or before or The different pathological parting recommended according to various tumour diagnosis and treatment specifications or guideline and advance stages tumour can be carried out later Therapeutic scheme, the tumour diagnosis and treatment specification or guideline include but is not limited to NCCN (the comprehensive cancer network hair of US National The various malignant tumour clinical practice guidelines of cloth) or ministry of Health of China promulgate malignant tumour diagnosis and treatment specification.

In the present invention, PD-1 antibody or its antigen-binding fragment and mek inhibitor are (such as 28 in identical treatment cycle It is a treatment cycle) combine for treating tumour, mek inhibitor administration synchronous with PD-1 antibody or its antigen-binding fragment Or it is administered before PD-1 antibody or its antigen-binding fragment or after PD-1 antibody or its antigen-binding fragment；Identical In dosage period, mek inhibitor is administered within a continuous period, and preferably 1-21 days successive administrations are continuously given for 1-28 days Medicine, the continuous period can be at any one time points of a treatment cycle, when the preferred therapeutic period starts；Phase With dosage period in, PD-1 antibody or its antigen-binding fragment administration frequency can be 2 weeks/time, 3 weeks/time or 4 weeks/time, can also To be administered at the 1st day of a treatment cycle and the 15th day.

The present invention is a kind of administration mode about " joint ", refers to and gives at least one dosage within the certain time time limit The mek inhibitor of PD-1 antibody or its antigen-binding fragment and at least one dosage, two of them substance all show that pharmacology is made With.The time limit can be in a dosage period, in preferably 4 weeks, in 3 weeks, in 2 weeks, in 1 week or 24 hours with It is interior, within more preferable 12 hours.PD-1 antibody or its antigen-binding fragment and mek inhibitor can simultaneously or sequentially be given.This The kind time limit includes such treatment, wherein giving PD-1 antibody or its antigen knot by identical administration route or different way of administration Close segment and mek inhibitor.United administration mode of the present invention, which is selected from, is administered simultaneously, independently prepares and is total to administration or only It on the spot prepares and is administered in succession.

In the present invention, the invention further relates to drug in purposes, wherein the PD-1 antibody or its antigen knot Close segment dosage rate be once a day, two times a day, three times per day, weekly, two weeks once, once in three weeks, January Once, the dosage rate of the mek inhibitor be once a day, two times a day, three times per day, weekly, two weeks once, three Zhou Yici, once a month.

In particularly preferred embodiments, the dosage rate of the PD-1 antibody or its antigen-binding fragment was two Mondays Secondary, the dosage rate of the mek inhibitor is once a day.

In a preferred embodiment of the present invention, the PD-1 antibody or its antigen-binding fragment are given in a manner of injecting Medicine, such as subcutaneous or intravenous injection, need the form that PD-1 antibody or its antigen-binding fragment are configured to injectable before injection.It is special The injectable forms of not preferred PD-1 antibody or its antigen-binding fragment are injection or freeze-dried powder, and it includes PD-1 antibody Or its antigen-binding fragment, buffer, stabilizer, optionally also contain surfactant.Buffer can be selected from acetate, lemon One or more of hydrochlorate, succinate and phosphate.Stabilizer can be selected from sugar or amino acid, preferably disaccharides, such as sugarcane Sugar, lactose, trehalose, maltose.Surfactant is selected from Crodaret, fatty acid glyceride, polyoxyethylene Sorbitan carboxylic esters, the preferably described polyoxyethylene sorbitan carboxylic ester are polysorbate 20,40,60 or 80, optimal Select polysorbate 20.The injectable forms of highly preferred PD-1 antibody or its antigen-binding fragment include PD-1 antibody or it is anti- Former binding fragment, acetate buffer, trehalose and polysorbate 20.

United administration mode of the present invention, which is selected from, to be administered simultaneously, independently prepare and is total to administration or independently prepare simultaneously Administration in succession.

United administration route of the present invention is selected from oral administration, parenteral, percutaneous dosing, described parenterally to give Medicine includes but is not limited to be injected intravenously, be subcutaneously injected, intramuscular injection.

The present invention provides above-mentioned immunotherapeutic agent and combines drug of the above-mentioned mek inhibitor as treatment tumour, wherein described Immunotherapeutic agent is selected from PD-1 antibody or its antigen-binding fragment.

In the present invention, a kind of method for the treatment of is provided, including applies above-mentioned immunotherapeutic agent and above-mentioned MEK to patient Inhibitor, wherein the immunotherapeutic agent is selected from PD-1 antibody or its antigen-binding fragment.

The present invention also provides a kind of medicine sleeve group or a kind of medicine package boxes, wherein containing mek inhibitor above-mentioned With PD-1 antibody or its antigen-binding fragment.

The present invention also provides a kind of pharmaceutical compositions, include a effective amount of PD-1 antibody above-mentioned or its antigen binding fragment Section and mek inhibitor and one or more pharmaceutical excipients, diluent or carrier.

Detailed description of the invention

Situation is assessed in the treatment of each subject of Fig. 1

Detailed description of the invention

One, term

In order to be easier to understand the present invention, certain technical and scientific terms are defined in detail below.Except obviously at this It is separately explicitly defined at it in file, otherwise all other technical and scientific term used herein all has belonging to the present invention The normally understood meaning of the those skilled in the art in field.

Term " humanized antibody (humanized antibody) ", also referred to as CDR grafted antibody (CDR-grafted Antibody), refer to the antibody variable region frame that the CDR sequence of mouse is transplanted to people, i.e., different types of human germline antibody The antibody generated in frame sequence.Chimeric antibody can be overcome due to carrying a large amount of murine protein ingredients, so that induction is strong Antibody variable antibody response.Such frame sequence can be from public DNA database or public affairs including germline antibody gene sequences The bibliography opened obtains.As people's heavy chain and the germline DNA sequence dna of light-chain variable region gene can be in " VBase " human germ line sequences Database (can get) in internet www.mrccpe.com.ac.uk/vbase, and in Kabat, E.A. et al., 1991Sequences of Proteins of Immunological Interest is found in the 5th edition.At the present invention one In preferred embodiment, the CDR sequence of the PD-1 humanized antibody mouse is selected from SEQ ID NO:1, and 2,3,4,5,6.

Term " antigen-binding fragment ", refers to the Fab segment with antigen-binding activity, Fab ' segment, 2 segment of F (ab '), And the Fv segment sFv segment in conjunction with people PD-1；SEQ ID NO:1 to SEQ ID is selected from comprising antibody of the present invention One or more CDR regions in NO:6.Fv segment contains antibody heavy chain variable region and light chain variable region, but does not have constant region, and Minimum antibody fragment with whole antigen binding sites.Generally, Fv antibody also includes more between VH and VL structural domain Peptide linker, and structure needed for being capable of forming antigen binding.Two antibody variable regions can also be connected with different attachments At a polypeptide chain, referred to as single-chain antibody (single chain antibody) or scFv (sFv).Term of the invention " with PD-1 combination ", referring to can interact with people PD-1.Term " antigen binding site " of the invention refer to it is discontinuous on antigen, by The three-dimensional space site of antibody or antigen-binding fragment identification of the present invention.

Term " immunotherapy " refers to that immunotherapy is that disease is treated using immune system, refers mainly to pass through in the present invention The immunogenicity of tumour cell and the sensibility of pairing effect cell killing, excitation and enhancing antitumor immunity of organism response are improved, And in application immunocyte and effector molecule infusion host, collaboration body immune system killing tumour inhibits tumour growth.

Specific embodiment

With reference to embodiments for further describing the present invention, but these embodiments are not intended to limit the scope of the invention.

Embodiment 1: -1 Antybody therapy advanced solid tumor clinical research of compound (1) tosilate combined PD

1, Subject antibodies and compound

PD-1 antibody is H005-1, heavy, light chain sequence by the preparation of method disclosed in WO2015085847, its corresponding code name Column such as SEQID NO:7 and SEQID NO:8 in the present invention.Lot number: it is spare to be made into 20mg/ml for P1512,200mg/ branch.

Compound (1) tosilate (hereinafter referred to as compound A) can press disclosed method system by WO2016155473 It is standby, tablet is made, specification is 0.125mg/ piece, 0.5mg/ piece, 2mg/ piece.

2, enrolled subject's standard

(1) age 18-70 years old；

(2) be diagnosed as advanced solid tumor and without respective standard treatment or treatment it is invalid；

(3) there are RAS or RAF Positive mutants；

(4) ECOG scores 0-1 points；

(5) life expectancy at least three moon；

(6) must have enough marrow, liver, kidney, lung and cardiac function.

3, medication

Enrolled subject receives compound A single-dose first, elutes 7-10 days after oral, subsequently into successive administration rank Section, once a day, continuous 28 days are a cycle to compound A tablet；The the 1st and the 15th day of the 1st period of successive administration receives H005-1, dosage are 200mg/ times.It is administered in combination until progression of disease, toxicity do not tolerate, subject actively cancels notice of knowing Book or researcher judge that subject is no longer appropriate for continuing to receive treatment.

Up to now, 10 clinics for participating in dosing phase with recurrence or metastatic colorectal carcinoma subject are tried It tests.Subject receives compound A (0.125mg/qd, 0.25mg/qd and 0.5mg/qd) and PD-1 antibody (200mg) is combined and controlled It treats.It shares 5 subjects and has dropped out the research.2 subject's progression of disease, 1 subject occur to treat unrelated SAE, 1 Example brain metastes subject (not meeting inclusion criteria) and 1 subject's (0.5mg/qd dosage group) undergo dose-limiting toxicity. Still have 5 subjects in research treatment, wherein 2 subjects are PR (respectively 11.5 months and 7 months), in addition 2 by Examination person is stable disease (respectively 9 months and 1 month).1 subject will receive tumor evaluation.The treatment of each subject is commented It is as shown in Figure 1 to estimate situation.

The gene mutation situation of each subject is as follows:

The genotype of subject

Sequence table

<110>Hengrui Medicine Co., Ltd., Jiangsu Prov.

<120>PD-1 antibody and mek inhibitor combine the purposes in the drug of preparation treatment tumour

<160> 10

<170> SIPOSequenceListing 1.0

<210> 1

<211> 5

<212> PRT

<213>source of mouse (Mus musculus)

<400> 1

Ser Tyr Met Met Ser

1 5

<210> 2

<211> 17

<212> PRT

<213>source of mouse (Mus musculus)

<400> 2

Thr Ile Ser Gly Gly Gly Ala Asn Thr Tyr Tyr Pro Asp Ser Val Lys

1 5 10 15

Gly

<210> 3

<211> 7

<212> PRT

<213>source of mouse (Mus musculus)

<400> 3

Gln Leu Tyr Tyr Phe Asp Tyr

1 5

<210> 4

<211> 11

<212> PRT

<213>source of mouse (Mus musculus)

<400> 4

Leu Ala Ser Gln Thr Ile Gly Thr Trp Leu Thr

1 5 10

<210> 5

<211> 7

<212> PRT

<213>source of mouse (Mus musculus)

<400> 5

Thr Ala Thr Ser Leu Ala Asp

1 5

<210> 6

<211> 9

<212> PRT

<213>source of mouse (Mus musculus)

<400> 6

Gln Gln Val Tyr Ser Ile Pro Trp Thr

1 5

<210> 7

<211> 443

<212> PRT

<213>artificial sequence (Artificial Sequence)

<220>

<221> PEPTIDE

<222> (1)..(443)

<223>sequence of heavy chain

<400> 7

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Met Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Thr Ile Ser Gly Gly Gly Ala Asn Thr Tyr Tyr Pro Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gln Leu Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Thr Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro

210 215 220

Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro

225 230 235 240

Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr

245 250 255

Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn

260 265 270

Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg

275 280 285

Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val

290 295 300

Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser

305 310 315 320

Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys

325 330 335

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu

340 345 350

Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe

355 360 365

Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu

370 375 380

Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe

385 390 395 400

Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly

405 410 415

Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr

420 425 430

Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 8

<211> 214

<212> PRT

<213>artificial sequence (Artificial Sequence)

<220>

<221> PEPTIDE

<222> (1)..(214)

<223>sequence of light chain

<400> 8

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Leu Ala Ser Gln Thr Ile Gly Thr Trp

20 25 30

Leu Thr Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Thr Ala Thr Ser Leu Ala Asp Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Val Tyr Ser Ile Pro Trp

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 9

<211> 116

<212> PRT

<213>artificial sequence (Artificial Sequence)

<220>

<221> PEPTIDE

<222> (1)..(116)

<223>heavy chain variable region

<400> 9

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Met Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Thr Ile Ser Gly Gly Gly Ala Asn Thr Tyr Tyr Pro Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gln Leu Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Thr Val

100 105 110

Thr Val Ser Ser

115

<210> 10

<211> 107

<212> PRT

<213>artificial sequence (Artificial Sequence)

<220>

<221> PEPTIDE

<222> (1)..(10)

<223>light chain variable region

<400> 10

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Leu Ala Ser Gln Thr Ile Gly Thr Trp

20 25 30

Leu Thr Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Thr Ala Thr Ser Leu Ala Asp Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Val Tyr Ser Ile Pro Trp

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

Claims

1. immunotherapeutic agent and mek inhibitor combine the purposes in the drug of preparation treatment tumour, which is characterized in that described to exempt from Epidemic disease therapeutic agent is selected from PD-1 antibody or its antigen-binding fragment, the light chain variable region of the PD-1 antibody or its antigen-binding fragment Include LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6 respectively；It is described The heavy chain variable region of PD-1 antibody or its antigen-binding fragment includes respectively such as SEQ ID NO:1, SEQ ID NO:2 and SEQ ID HCDR1, HCDR2 and HCDR3 shown in NO:3；The mek inhibitor is selected from AZD-8330, GDC-0623, CI-1040, WX- 554、TAK-733、HL-085、BI-847325、CEP-1347、Binimetinib、Pimasertib、Cobimetinib、PD- 0325901, RO-5126766, Trametinib, Refametinib, AS-703988, E-6201, Selumetinib or as follows Compound represented (1) or its officinal salt,

2. purposes according to claim 1, which is characterized in that the PD-1 antibody or its antigen-binding fragment are selected from PD-1 Humanized antibody.

3. purposes according to claim 2, wherein the light-chain variable sequence of the anti-PD-1 humanized antibody is such as SEQ Sequence shown in ID NO:10 or its variant, the variant preferably has the amino acid of 0-10 to change in light chain variable region, more excellent It is selected as the amino acid variation of A43S；Weight chain variabl area sequence is the sequence as shown in SEQ ID NO:9 or its variant, the variant It is preferred that there is the amino acid of 0-10 to change in heavy chain variable region, the amino acid variation of more preferably G44R.

4. purposes according to claim 1-3, which is characterized in that PD-1 humanized antibody light chain's sequence For the sequence as shown in SEQ ID NO:8, sequence of heavy chain is the sequence as shown in SEQ ID NO:7.

5. purposes according to claim 1-4, which is characterized in that the tumour is selected from malignant tumour, benign swollen Tumor；It is swollen that the malignant tumour is selected from carcinoma, sarcoma, myeloma, leukaemia, lymthoma, melanoma, incidence Tumor, brain tumor, peritoneal cancer, mixed type tumour, children malignant tumors.

6. purposes according to claim 1-5, which is characterized in that the tumour is selected from Hodgkin lymphoma, non- Hodgkin lymphoma, prostate cancer, cancer of pancreas, lung cancer, the cancer of the esophagus, liver cancer, cholangiocarcinoma, breast cancer, colorectal cancer, gastric cancer, kidney The unknown tumour of cancer, acute myelogenous lymphocytic leukemia, myelodysplastic syndrome, glioma, original site.

7. purposes according to claim 1-6, which is characterized in that the tumour is mediated and/or expressed by PD-1 PD-L1。

8. purposes according to claim 1-7, which is characterized in that the tumour be selected from RAS mutant tumours, RAF mutant tumours, RAS/RAF/MEK access ectype tumour, RAS/RAF access ectype tumour.

9. purposes according to claim 1-8, which is characterized in that the tumour is selected from Advanced cancers, recurrence Refractory neoplasm, through chemotherapeutic drug therapy failure and/or recurrent tumor, through radiotherapy failure and/or recurrent tumor, through targeted drug Treatment failure and/or recurrent tumor, through immunization therapy failure and/or recurrent tumor.

10. -9 described in any item purposes according to claim 1, which is characterized in that the PD-1 antibody or its antigen binding Segment dosage is selected from 1-10mg/kg, preferably 3mg/kg, 4mg/kg, 5mg/kg.

11. -10 described in any item purposes according to claim 1, which is characterized in that the PD-1 antibody or its antigen binding Segment dosage is selected from 50-600mg, preferably 200mg, 400mg, 600mg.

12. -11 described in any item purposes according to claim 1, which is characterized in that the mek inhibitor dosage is selected from 0.01- 500mg, preferably be selected from 0.1mg, 0.25mg, 0.5mg, 0.75mg, 1mg, 2mg, 3mg, 4mg, 5mg, 6mg, 7mg, 8mg, 9mg, 10mg、11mg、12mg、12.5mg、15mg、17.5mg、20mg、22.5mg、25mg、30mg、45mg、50mg、60mg、70mg、 75mg、80mg、90mg、100mg、125mg、150mg、175mg、200mg、225mg、250mg、275mg、300mg、400mg、 500mg, more preferable 0.25mg, 0.5mg, 1mg, 2mg, 3mg, 4mg, 10mg, 15mg, 20mg, 30mg, 45mg, 50mg, 60mg, 75mg、100mg。

13. -12 described in any item purposes according to claim 1, which is characterized in that the officinal salt of the compound (1) selects From tosilate.

14. a kind of medicine package box, which is characterized in that include mek inhibitor described in claim 1-13 any one and PD-1 antibody or its antigen-binding fragment.

15. a kind of pharmaceutical composition, which is characterized in that include a effective amount of PD-1 described in claim 1-13 any one Antibody or its antigen-binding fragment and mek inhibitor and one or more pharmaceutical excipients, diluent or carrier.